Methods for immune or hematological stimulation, tumour formation or growth inhibition or prevention of malignant tumour, symptoms of malignant tumours or symptoms related to treatment of malignant tumours

FIELD: medicine, pharmaceutics.

SUBSTANCE: group of inventions refers to introducing dairy fat or an analogue thereof, optionally with at least one additional therapeutic factor, preferentially with lactoferrin or lactoferrin containing a metal ion, preferentially lactoferrin containing iron, preferentially bovine lactoferrin containing iron or a functional version thereof containing metal ions or a functional fragment to inhibit the tumour formation or growth, to maintain or improve one or more parameters, such as leukocyte count, erythrocyte count, or myelocyte count, to reduce the manifestations of cachexia, mucositis and anemia, to stimulate the immune system and to treat or prevent a malignancy and malignant symptoms, and side effects of treating the malignant tumour. Methods and therapeutic use according to the invention may be implemented by the use of a diet (in the form of food products or food additives), nutrient or pharmaceutical composition. There are also presented compositions applicable in the methods according to the invention.

EFFECT: group of inventions provides the higher clinical effectiveness or prevention of the malignant tumour or its symptoms.

39 cl, 7 tbl, 21 ex

 

The scope of the invention

The present invention relates to methods of immune or hematological stimulation, inhibition of the formation or growth of the tumor and the treatment or prevention of malignant tumors, the symptoms of a malignant tumor or symptoms associated with the treatment of malignant tumors through the introduction of milk fat or similar milk fat, optionally, at least one additional drug, such as an antitumor agent, preferably selected from lactoferrin (including iron-lactoferrin). Methods and therapeutic use according to the invention can be realized by applying the diet (in the form of food or supplements), nutraceutical or pharmaceutical composition. Also provided compositions useful in the methods according to the invention.

Background of the invention

Milk is rich in biological fluid that provides nourishment during rapid growth and development in infancy. Therefore, in addition to the substrates necessary for the development of the baby, it contains a lot of growth regulators.

Health benefits from drinking milk on the risk of development of malignant tumors is the subject of research and extensive discussions. It was reported that consumption of dairy products is mswb with high fat content (Larsson, et al., 2005) or milk with low fat content (Ma, et al., 2001 and Goodman, et al., 2002) may reduce the risk of certain cancers, including cancer of the colon, rectum and cancer of the ovaries. On the other hand, other studies have described that high consumption of milk with low fat increases the risk of some malignancies (Larsson, et al., 2006). In the literature review reported that the evidence for the consumption of dairy products with risk of ovarian cancer is limited and contradictory (Schulz, et al., 2004). With reduced breast cancer risk (Shin, et al., 2002 and Bradlow and Sepkovic, 2002) was connected as whole milk, and milk with low fat content, whereas in other studies they have been described as risk factors (Knekt, et al., 1996 and Gaard, et al., 1995). Milk, low-fat contributes to the development of induced carcinogens of mammary tumors in rats (Qin, et al., 2004), as well as milk with low fat content was associated with an increased risk of prostate cancer (Tseng, et al., 2005 and Veierod et al, 1997). In one large prospective study published that drinking milk as an adult has a tendency to negative correlation with the incidence of breast cancer (Hjartaker, et al., 2001), whereas in recent reviews concluded that between elevated or reduced risk of malignant is th tumors and high consumption of dairy products as low and high-fat regularities no (Moorman and Terry, 2004 and Parodi, 2005). In conclusion, it should be noted that the epidemiological evidence regarding the risk of development of malignant tumors and benefits of drinking milk are contradictory.

Milk fat contains a number of components, including conjugate linoleic acid (CLA), sphingomyelin, butyric acid, vaccinology acid, fatty acids with branched-chain lipids ethers, β-carotene and vitamins A and D, which were examined to evaluate their anticancer potential (reviewed in Parodi, 1999 and Parodi, 1997). For example, according to published data, CIS-9, TRANS-11 (c9, t11) CLA isomer and its predecessor Aksenova acid inhibit the growth of tumor cell lines (Miller, et al., 2003 and O'shea, et al., 2000). According to published data, the diet with CLA, increasing apoptosis, reduces the incidence of colon cancer in rats, which were administered 1,2-dimethylhydrazine (Kim and Park, 2003). Other studies have described that the diet with CLA inhibits the initiation of skin carcinogenesis in mice induced by 7,12-dimethyl-Benz[a]anthracene (DMBA) (Ha et al., 1987) and neoplasia of the gastric cardia in mice induced by benzo[a]pyrene (Chen, et al., 2003). Described that feeding vaccinology acid and c9, t11 CLA in the form of fat oils inhibited the development of carcinomas of the mammary glands in rats (Corl et al., 2003, Banni, et al., 2001). It was reported that CLA inhibits angiogenesis, which may contribute to its effectiveness as chemopreventive funds (Masso-Welch, et al., 2002).

In contrast, other studies have described that the diet with CLA individually or in combination with isolate soy protein did not inhibit the growth and development of tumor cells of the prostate gland in rats in vivo (Cohen, et al., 2003) and the development of aberrant crypt foci in the colon of male rats Sprague-Dawley, induced azoxymethane (Ealey et al., 2001). In addition, published that the mass Hepatol rats was significantly higher than in rats, which were kept on a diet with CLA (Yamasaki, et al., 2001). In epidemiological studies published that the occurrence of breast cancer was not associated with consumption of food products containing CLA (Voorrips et al., 2002). As in the case of epidemiological studies of consumption of milk fat people, the results of various studies CLA and antitumor activity are contradictory.

Previously published that the tumor is not very sensitive to chemotherapy in all cases. For example, the effectiveness of chemotherapy for subjects with malignant tumors depends on the tumor type, the type and dose of drugs used for treatment, the mechanisms through which work drugs is a war funds and schemes of treatment.

In this area it is known that malignant tumors are distinguished by their sensitivity to chemotherapy from usually and often sensitive (e.g., lymphoma, acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), Hodgkin's disease, intermediate and high grade non-Hodgkin's lymphoma, such as diffuse both lymphoma, lymphoma Berkata, lymphoblastic lymphoma, horiokartsinoma, embryonal tumors, multiple myeloma, oat cell bronchial carcinoma, testicular carcinoma, Ewing sarcoma, Wilm tumor, skin cancer), where you can achieve a complete clinical recovery, to more sustainable (bladder cancer, esophageal cancer, non-small cell lung cancer, liver carcinoma, carcinoma, kidney carcinoma, pancreatic, head and neck cancer, cervical carcinoma, liver carcinoma, lung carcinoma, which are not oat cell). The previously described that tumor EL-4, constituting more than 0.3 cm in diameter, become completely insensitive to immunotherapy and antiangiogenic treatment (Kanwar, et al, 1999, Sun et al., 2001) and chemotherapy (Kanwar et al. - WO 2006/054908).

In addition, treatment of malignant tumors by radiation therapy, or surgery, or chemotherapy, or by other means, often causes or exacerbates assosiated with her SIM is Toms or disease. For example, subjects with malignant tumors, treatment, often cachectin, because chemotherapy can cause inflammation of the mucous membrane of the small intestine associated with apoptosis in the crypts, which precedes atrophy of intestinal villi (Keefe, et al., 2000).

Thus, it is desirable to develop an improved method of inhibiting the formation or growth of a tumor or reducing the symptoms or severity of diseases associated with malignant tumor, or treatment of malignant tumors with the use of milk fat, optionally with one or more additional drugs, or at least to give people the right choice.

The invention

Thus, one aspect of the invention relates to a method of inhibiting the formation of a tumor, inhibiting tumor growth, inhibiting metastasis of a tumor or the treatment or prevention of a malignant tumor in the subject, where the method comprises the separate, simultaneous or sequential introduction to the needy in this subject an effective amount of milk fat or similar milk fat and one or more drugs, such as one or more anticancer agents, where preferably one or more anticancer which means selected from anticancer dietary factors, chemotherapy, immunotherapy, hematopoietic funds protivokahekticalkie funds or protivomoskitnykh funds, more preferably one or more drugs are lactoferrin (Lf).

Another aspect of the invention relates to a method of stimulating the immune system of a subject, where the method includes the introduction to the needy in this subject an effective amount of milk fat or similar milk fat. In one embodiment, the implementation method of stimulating the immune system includes separate, simultaneous or sequential administration to the subject of milk fat and one or more anticancer agents, where preferably one or more anticancer agents selected from anticancer dietary factors, chemotherapeutic agents, immunotherapy, hematopoietic funds protivokahekticalkie funds or protivomoskitnykh funds, where one or more drugs more preferably represent lactoferrin.

In one embodiment, the implementation of introduction increases the production of cytokines by Th1 and Th2 in the tumor of the subject. In one embodiment, the implementation of introduction increases the production of cytokines by Th1 and Th2 in the intestine of a subject. In one embodiment, the implementation of the centuries is typically increases the level of Th1 cytokines and Th2 in the circulatory system of the subject. In one embodiment, the implementation of the introduction raises the subject of the antitumor immune response.

Another aspect of the invention relates to a method of inducing apoptosis in a subject, where the method includes introducing an effective amount of milk fat or similar milk fat needy in this subject. In one embodiment, the implementation method of promoting apoptosis in need of this subject includes separate, simultaneous or sequential administration to the subject of milk fat and at least one antineoplastic agents, where at least one antitumor agent is preferably selected from anticancer dietary factors, chemotherapeutic agents, immunotherapy, hematopoietic funds protivokahekticalkie funds or protivomoskitnykh funds, where preferably at least one additional drug is a lactoferrin. In one embodiment, the implementation of apoptosis represents the apoptosis of tumor cells.

Another aspect of the invention relates to a method of inhibiting angiogenesis in a subject, where the method includes introducing an effective amount of milk fat or similar milk fat needy in this subject. In one embodiment, the implementation of the method of inhibiting Angie is ESA we need in this subject includes separate, simultaneous or sequential administration to the subject of milk fat and at least one antineoplastic agents, where preferably at least one antitumor agent selected from anticancer dietary factors, chemotherapeutic agents, immunotherapy, hematopoietic funds protivokahekticalkie funds or protivomoskitnykh funds, where preferably at least one additional drug is a lactoferrin. In one embodiment, the implementation of angiogenesis is a tumor angiogenesis.

Another aspect of the invention relates to a method of treating or preventing anemia caused by low levels of hemoglobin or red blood cells, cachexia, mucositis or leukopenia, or maintain or improve one or more of the quantities of leukocytes, red blood cells and myeloid cells in the subject, where the method includes introducing an effective amount of milk fat or similar milk fat needy in this subject.

Another aspect of the invention relates to a method for maintaining or increasing the entity has one or more of the number of leukocytes, the number of red blood cells or the number of myeloid cells, where the method includes introduction to the subject of milk fat or similar dairy siraprapasiri with the introduction of the subject separately, simultaneously or sequentially at least one antineoplastic agents, where preferably at least one antitumor agent selected from anticancer dietary factors, chemotherapeutic agents, immunotherapy, hematopoietic funds protivokahekticalkie funds or protivomoskitnykh funds, where preferably at least one additional drug is a lactoferrin.

Another aspect of the invention relates to a method of treating or preventing the subject of anemia, where the method includes introduction to the subject of milk fat or similar milk fat, preferably with the introduction of the subject separately, simultaneously or sequentially at least one antineoplastic agents, where preferably at least one antitumor agent selected from anticancer dietary factors, chemotherapeutic agents, immunotherapy, hematopoietic funds protivokahekticalkie funds or protivomoskitnykh funds, where preferably at least one additional drug is a lactoferrin.

Another aspect of the invention relates to a method of treating or preventing cachexia in a subject, where the method involves rst is giving to the subject of milk fat or similar milk fat, preferably with the introduction of the subject separately, simultaneously or sequentially at least one antineoplastic agents, where preferably at least one antitumor agent selected from anticancer dietary factors, chemotherapeutic agents, immunotherapy, hematopoietic funds protivokahekticalkie funds or protivomoskitnykh funds, where preferably at least one additional drug is a lactoferrin.

Another aspect of the invention relates to a method of treating or preventing in a subject mucositis, where the method includes introduction to the subject of milk fat or similar milk fat, preferably with the introduction of the subject separately, simultaneously or sequentially at least one antineoplastic agents, where preferably at least one antitumor agent selected from anticancer dietary factors, chemotherapeutic agents, immunotherapy, hematopoietic funds protivokahekticalkie funds or protivomoskitnykh funds, where preferably at least one additional drug is a lactoferrin.

Another aspect of the invention relates to a method of inhibiting the formation of a tumor, inhibiting tumor growth, inhib the regulation of metastasis of the tumor or the treatment or prevention of a malignant tumor in the subject, where the method includes the introduction of the subject of fractions of milk fat, such as phospholipid fraction, fraction solid milk fat fraction soft milk fat, sphingolipids faction, faction shell globules of milk fat, phospholipid fraction, or complex lipid fraction, or a combination of any two or more of them, optionally, at least one additional drug, where preferably at least one additional drug selected from anticancer dietary factors, chemotherapeutic agents, immunotherapy, hematopoietic funds protivokahekticalkie funds or protivomoskitnykh funds, where preferably at least one additional drug the tool is a lactoferrin (Lf).

Another aspect of the invention relates to a method for increasing autochemistry subject for treatment of a malignant tumor, where the method includes introduction to the subject of milk fat or similar milk fat, preferably with the introduction of the subject separately, simultaneously or sequentially at least one antineoplastic agents, where at least one antitumor agent is preferably selected from anticancer dietary factors, chemotherapeutic agents, immuno rapeutically funds hematopoietic funds protivokahekticalkie funds or protivomoskitnykh funds, where preferably at least one additional drug is a lactoferrin.

Another aspect of the invention relates to a method for increasing the sensitivity of a tumor of a subject to treatment of a malignant tumor, where the method includes introduction to the subject of milk fat or similar milk fat, preferably with the introduction of the subject separately, simultaneously or sequentially at least one antineoplastic agents, where preferably at least one antitumor agent selected from anticancer dietary factors, chemotherapeutic agents, immunotherapy, hematopoietic funds protivokahekticalkie funds or protivomoskitnykh funds, where preferably at least one additional drug is a lactoferrin.

Another aspect of the invention relates to a method for accelerating the recovery of the subject being treated for a malignant tumor, where the method includes introduction to the subject of milk fat or similar milk fat, preferably with the introduction of the subject separately, simultaneously or sequentially at least one antineoplastic agents, where predpochtitelno at least one antitumor agent selected from anticancer dietary factors, immunotherapy hematopoietic funds protivokahekticalkie funds or protivomoskitnykh funds, where preferably at least one additional drug is a lactoferrin.

Another aspect of the invention relates to the use of milk fat in the production of compositions for purpose, as described herein, where preferably the composition includes at least one additional drug or introduced separately, simultaneously or sequentially with at least one additional drug, where preferably at least one additional drug is an anticancer agent, where an antitumor agent preferably selected from anticancer dietary factors, chemotherapeutic agents, immunotherapy, hematopoietic funds protivokahekticalkie funds or protivomoskitnykh funds, where preferably at least one additional drug is a lactoferrin.

Another aspect of the invention relates to the use of milk fat, optionally at least one additional drug in the manufacture of compositions for purpose, as described in the present on the document.

Another aspect of the invention relates to the use of milk fat and at least one additional drug in the manufacture of compositions for purpose, as described herein, where the composition is to provide separate, simultaneous or sequential introduction of milk fat and at least one additional drug.

Another aspect of the invention relates to the use of milk fat and at least one additional drug in the manufacture of compositions for purpose, as described herein, where the milk fat is administered separately, simultaneously or sequentially with an additional drug.

Another aspect of the invention relates to the use of milk fat and at least one additional drug in the manufacture of compositions for purpose, as described herein, where the milk fat are for separate, simultaneous or sequential introduction of additional drug.

Another aspect of the invention relates to a composition, which includes mainly contains or consists of milk fat and one or more, two or more or three or more additional drugs.

Another aspect of the image is possible only applies to the product, which includes, mainly contains or consists of milk fat and one or more, two or more or three or more additional drugs, as a combined preparation for simultaneous, separate or sequential use for the purpose as described in this document.

The following options for implementation may treat any of the above aspects.

In preferred embodiments, the implementation of at least one drug is an anticancer agent. In preferred embodiments, the implementation of the antitumor agent is a lactoferrin.

In one embodiment, the implementation of lactoferrin is selected from the group comprising a lactoferrin polypeptide, a functional lactoferrin variant, a functional fragment of lactoferrin containing a metal ion lactoferrin functional variant containing a metal ion lactoferrin functional fragment containing the metal ion lactoferrin or a mixture of any two or more of them. In one embodiment, the implementation of lactoferrin is apolactoferrin. In another embodiment, the lactoferrin is a natural saturated iron lactoferrin. In another embodiment, the lactoferrin is an essentially completely is rich in iron lactoferrin.

Goals described in this document, include an objective selected from the inhibition of the formation of a tumor, inhibiting tumor growth, inhibiting metastasis of a tumor or of treating or preventing malignant tumors in need thereof of a subject, stimulating the immune system in in need thereof of a subject, increasing the production of cytokines by Th1 and Th2 in tumors in need thereof of a subject, increasing the production of cytokines by Th1 and Th2 in the intestine in need of this subject, enhancing the production of cytokines by Th1 and Th2 in the circulatory system we need in this subject, enhance antitumor immune response in in need thereof of a subject, stimulating apoptosis the need for this subject, stimulation of apoptosis of tumor cells in in need thereof of a subject, inhibiting angiogenesis in in need thereof of a subject, inhibiting angiogenesis in a tumor in need of this subject, maintain or improve one or more of the quantities of leukocytes, red blood cells or myeloid cells in in need thereof of a subject, treating or preventing anemia in need thereof of a subject, treating or preventing cachexia in in need thereof of a subject, treating or preventing mucositis in in need thereof of a subject, increasing autochemistry subject for treatment of evil is Aceitunas tumor, increase the sensitivity of a tumor of a subject to treatment of a malignant tumor and accelerate the recovery of the subject being treated for a malignant tumor.

In one embodiment, the implementation of at least one drug or anti-cancer agent selected from the group consisting of anticancer dietary factors, chemotherapeutic agents, immunotherapy, hematopoietic funds protivokahekticalkie funds or protivopokazanie tools.

In one embodiment, the implementation of anticancer food factor selected from vitamin D and analogues of vitamin D (including as non-limiting examples, i.e. which are discussed below), soy protein, one or more components of soybeans (including those discussed below), polyphenols, lycopene, wheat bran, flavanoids, Inositol, resveratrol, propolis, mushroom extract, anthocyanins, almonds, ginseng extract, hydrolyzed casein, and combinations thereof.

In one embodiment, the implementation of anticancer food factor selected from the group including anticancer foods and anticancer dietary components. Preferably administered one or more, two or more or three or more anticancer dietary factors.

In one embodiment, implemented what I anticancer foods can be a functional food products or their derivatives, which have anti-cancer properties, including those described in this document.

In one embodiment, the implementation of anticancer food component can be selected from those described in this document.

In one embodiment, the implementation of the method according to the invention includes the introduction of a composition mainly containing or consisting of milk fat and at least one additional drug, where at least one additional drug is preferably selected from lactoferrin, a functional lactoferrin variant, a functional fragment of lactoferrin containing a metal ion lactoferrin functional lactoferrin variant containing a metal ion, a functional fragment of lactoferrin containing metal ion, or a mixture thereof. Preferably, the composition mainly contains or consists of one or more, two or more or three or more anticancer dietary factors.

In one of the embodiments of the application of the invention composition obtained for inhibition of the formation of a tumor in a subject, inhibiting growth of a tumor in a subject, inhibiting metastasis of a tumor in a subject, treating or preventing a malignant tumor in a subject, stimulate immune with the system of the subject, increase the production of cytokines by Th1 and Th2 in the tumor of the subject, increase the production of cytokines by Th1 and Th2 in the intestine of the subject, increase the production of cytokines by Th1 and Th2 in the circulatory system of the subject, enhance antitumor immune response in the subject, stimulation of apoptosis in the subject, stimulation of apoptosis of tumor cells in a subject, inhibiting angiogenesis in a subject, inhibiting angiogenesis in a tumor in the subject, maintain or improve one or more of the quantities of leukocytes, red blood cells or myeloid cells in a subject, treating or preventing anemia in need thereof of a subject, treating or preventing cachexia in need of this subject, of treating or preventing mucositis in in need thereof of a subject, treating or preventing radiation in need of this subject, the gain response of a subject to treatment of a malignant tumor, increased autochemistry subject for treatment of a malignant tumor or accelerate the recovery of a subject undergoing treatment for malignant tumors.

In one embodiment, the implementation of the introduction is an oral, local or parenteral administration.

In one of the embodiments the subject suffers from a malignant tumor or susceptible to malignant tumors; was cured the e but he had a relapse or it susceptible to relapse; has a tumor that is resistant to treatment with chemotherapy, radiotherapy, antiangiogenic or immunotherapy agent; or he has previously undergone surgical intervention, surgical intervention with poor outcome, or were unsuccessful treatment with chemotherapy, radiotherapy, anti-angiogenic or immunotherapy agent.

In one embodiment, the implementation of the milk fat is selected from milk lipids, lipid fractions of milk, hydrolysates of milk lipids and hydrolyzed fractions of milk lipids. Preferred dairy fats are fats milk, especially fats in cow's milk.

In other embodiments, the implementation of the milk fat is the fat of the milk of any mammal, including as non-limiting examples of the milk fat sheep, goats, pigs, mice, Asian water Buffalo, camel, Yak, horse, donkey, llama or person.

In one embodiment, the implementation of lactoferrin is a lactoferrin any mammal, including as non-limiting examples of lactoferrin sheep, goats, pigs, mice, Asian water Buffalo, camel, Yak, horse, donkey, llama, or person. Preferably lactoferrin is a bovine lactoferrin.

In one embodiment, about what westline lactoferrin is apolactoferrin. In one embodiment, the implementation of lactoferrin, a functional lactoferrin variant or functional fragment of lactoferrin does not contain metal ions. In one embodiment, the implementation of lactoferrin or a functional variant or functional fragment saturated with the metal ion of at least about 5, 10 or 20% in the stoichiometric ratio.

In one embodiment, the implementation of the metal ion is a metal ion selected from the group comprising aluminum, bismuth, copper, chromium, cobalt, gold, iron, manganese, osmium, platinum, ruthenium and zinc ions, or any combination of any two or more of them or other ions that will be specific to interact with the binding of metal ions in lactoferrin. Preferably the metal ion is an iron ion.

In one embodiment, the implementation of lactoferrin, a functional lactoferrin variant or functional fragment of lactoferrin are involved in non-specific binding of ions. Ions that may nonspecific contact lactoferrin, a functional variant of lactoferrin or a functional fragment of lactoferrin, preferably selected from aluminum, calcium, bismuth, copper, chromium, cobalt, gold, iron, manganese, osmium, platinum, ruthenium, selenium and zinc ions or any combination of any two is whether more of them. Ion can be any ion or mixture of ions, which are nonspecific contact with lactoferrin, a functional variant of lactoferrin or a functional fragment of lactoferrin, preferably ion calcium and selenium.

In one of the embodiments containing a metal ion lactoferrin or containing a metal ion functional variant or functional fragment saturated with the metal ion of at least about 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 99,5 or 100% of the stoichiometric value.

In one of the embodiments containing a metal ion lactoferrin or containing a metal ion functional variant or functional fragment saturated with the metal ion of at least about 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195 or 200% of the stoichiometric ratio.

In one of the embodiments the composition is suitable herein include from about 2 grams to about 210 grams of milk fat or similar milk fat and from about 0.1 grams to about 210 grams of one or more anticancer agents. In one of the embodiments the composition includes approximately 2, 4, 6, 8, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175 180, 185, 190, 195, 200, 205 or 210 grams of milk fat or similar milk fat and suitable ranges, you can choose between any of these values. In one of the embodiments the composition includes approximately 2, 4, 6, 8, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, 200, 205 or 210 grams of one or several anticancer agents, and suitable ranges, you can choose between any of these values. Preferably one or more anticancer agents selected from the group comprising lactoferrin, apolactoferrin, lactoferrin polypeptide, a functional lactoferrin variant, a functional fragment of lactoferrin containing metal ion lactoferrin, natural saturated iron lactoferrin, essentially fully saturated with iron lactoferrin, a functional variant containing a metal ion lactoferrin functional fragment containing the metal ion lactoferrin or a mixture. In a different implementation, the composition includes

(a) from about 2 grams to about 210 grams of milk fat or similar milk fat and from about 0.7 grams to about 70 grams of one or more anticancer agents;

(b) from about 35 grams to about 210 grams of milk fat or similar saying is knogo fat and from around 0.35 grams to about 210 grams of one or more anticancer agents;

(c) from about 10 grams to about 200 grams of milk fat or similar milk fat and from about 2.5 grams to about 70 grams of one or more anticancer agents;

(d) from about 10 grams to about 200 grams of milk fat or similar milk fat and from about 0.25 gram to about 5 grams of one or more anticancer agents;

(e) from about 15 grams to about 30 grams of milk fat or similar milk fat and from about 1 gram to about 6 grams of one or more anticancer agents; or

(f) from about 3 grams to about 8 grams of milk fat or similar milk fat and from about 0.1 gram to about 1 gram of one or more anticancer agents.

In one of the embodiments the method includes the introduction together with milk fat and blends containing metal ion lactoferrin and at least one containing a metal ion functional variant or functional fragment.

In one embodiment, the implementation of the milk fat and the additional drug, preferably lactoferrin, provide a synergistic therapeutic effect that is greater than the Indus the individual action of each, or greater than the sum of the actions of each of them individually. For example, there is a greater effect on inhibition of the formation or tumor growth, regression of tumor cytolytic effects, immune activation, the formation of cytokines Th1 and Th2, to maintain or improve the quantity of white blood cells, red blood cells or myeloid cells, treatment or prevention of anemia, cachexia, mucositis, or the response of the subject or of the tumor to the treatment. In one embodiment, the implementation of lactoferrin and anti-tumour food factors, in appropriate cases, reduce or increase the dose or duration of introduction when conducting simultaneous or sequential therapy of malignant tumors.

In one embodiment, the implementation of the method according to the invention further includes a separate, simultaneous or sequential administration of at least one method of treatment of malignant tumors.

In one embodiment, the implementation of the treatment of malignant tumors is an antineoplastic or antitumor therapy.

In one embodiment, the implementation of the milk fat, optionally, at least one additional pharmaceutical agent and at least one anticancer agent or anti-tumor therapy is administered separately is about, simultaneously or sequentially.

In one embodiment, the implementation of anticancer therapy as non-limiting examples selected from such treatments as surgery, chemotherapy treatments, methods of radiation therapy, the methods of hormonal therapy, biological methods of treatment/therapy, methods of cell therapy, methods antiangiogenic therapy, methods of cytotoxic therapy, vaccines, vaccines based on nucleic acids (e.g., nucleic acids expressing the antigen malignant tumors, such as DNA vaccines, including vaccines p185), treatment-based viruses (e.g., adeno-associated virus, lentivirus), methods of gene therapy, methods of treatment based on low molecular weight inhibitor, methods of therapy on the basis of nucleotides (for example, Rnci, antisense nucleotides, ribozymes etc), methods of treating an antibody-based, oxygen - ozone therapy, treatment by embolization and/or oily chemo-embolization. In one embodiment, the implementation of the anti-cancer agent includes one or more angiogenesis inhibitors.

In one embodiment, the implementation of the anti-cancer agent is a chemotherapeutic agent or immunotherapy agent. In one embodiment, the implementation is about at least one anti-cancer agent is a chemotherapeutic agent. Preferably the chemotherapeutic agent is selected from the disintegrator tubulin, intercalation DNA and mixtures thereof. In one embodiment, the implementation of the disintegrator tubulin, include as non-limiting examples of the disintegrators tubulin listed in the published international patent application WO 2006/054908, which is included in this document as a reference. In one embodiment, the implementation of intercalatory DNA include as non-limiting examples intercalatory DNA listed in the published international patent application WO 2006/054908, which is included in this document as a reference. In one embodiment, the implementation of the chemotherapeutic agent represents paclitaxel, doxorubicin, epirubicin, fluorouracil, cyclophosphamide or methotrexate.

In one embodiment, the implementation of the anti-cancer agent is an immunotherapy agent. Preferably immunotherapy product is expressing plasmid encoding costimulatory t cells, B7-1, costimulator T-cells or functionally related molecules, such as soluble chimeric molecule B7-Ig. In one embodiment, the implementation of the anti-cancer agent includes therapy by immune cells. Preferably the treatment is a is arapey dendritic cells.

In one embodiment, the implementation of the chemotherapeutic agent represents paclitaxel, doxorubicin, epirubicin, fluorouracil, cyclophosphamide or methotrexate.

In one embodiment, the implementation of the anti-cancer agent is an immunotherapy agent. Preferably immunotherapy represents an expression plasmid encoding costimulatory t cells, B7-1, costimulator T cells or functionally related molecules, such as soluble chimeric molecule B7-Ig.

In one embodiment, the implementation of the anti-cancer agent includes therapy by immune cells. Preferably the treatment is a dendritic cell therapy.

In one embodiment, the implementation of the anti-cancer agent includes one or more angiogenesis inhibitors.

In one embodiment, the implementation of at least one anti-tumor agent administered orally or parenterally, preferably by intravenous, intraperitoneal introduction or injection into the tumour.

In one embodiment, the implementation of the milk fat, optionally, at least one additional drug is administered daily for at least about 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 weeks before the introduction of the anti-Christ. opuholevogo funds or conducting cancer treatment.

In one embodiment, the implementation of the milk fat, optionally, at least one additional drug is administered for at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or 21 days, or within at least approximately 1, 2, 3, 4, 5, 6, 7 or 8 weeks, or for at least about 1, 2, 3, 4, 5 or 6 months before the introduction of anti-cancer agents and / or stage of cancer treatment.

In one embodiment, the implementation of the milk fat, optionally, at least one additional drug is administered for at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or 21 days, or within at least approximately 1, 2, 3, 4, 5, 6, 7 or 8 weeks, or for at least about 1, 2, 3, 4, 5, or 6 months after the introduction of anti-cancer agents or anti-tumour therapy.

In one embodiment, the implementation of the milk fat, optionally, at least one additional drug is administered at least once, including continuous oral administration within days or parenteral administration by infusion or by a combination of routes of administration with the treatment of malignant tumors or without it.

In one embodiment, the implementation of the op is hol or malignant tumor is a solid tumor, leukemia, lymphoma, multiple myeloma, hematopoietic tumors of lymphocytic origin, hematopoietic tumors of myeloid origin, carcinoma of the colon, breast cancer, melanoma, malignant tumor of the skin or lung cancer.

In one embodiment, the implementation of the tumor or malignancy is a leukemia, as non-limiting examples such as acute leukemia, acute lymphocytic leukemia, acute granulocytic leukemia, acute military leukemia, such as myeloblastic, promyelocytic, myelomonocytic, monocytic, erythroleucus and myelodysplastic syndrome, chronic leukemia, such as, but not limited to them, chronic miliitary leukemia, chronic granulocytic leukemia, chronic lymphocytic leukemia and hairy cell leukemia.

In one embodiment, the implementation of a tumor or a malignant tumor is a lymphoma, as non-limiting examples such as Hodgkin's disease and nahodkinskuju lymphoma.

In one embodiment, the implementation of a tumor or a malignant tumor includes hematopoietic tumor of myeloid origin, such as, but not limited to it, acute and chronic myelogenous leukemia, indolent multiple myeloma, non-secretory myeloma and osteosclerotic the Czech myeloma.

In one embodiment, the implementation of a tumor or a malignant tumor includes hematopoietic tumor lymphocytic origin, including leukemia, acute and chronic lymphocytic leukemia, acute and chronic lymphoblastic leukemia, B-cell lymphoma, T-cell lymphoma, lymphoma Bernita.

In one embodiment, the implementation of a tumor or a malignant tumor includes hematopoietic tumor of B-lymphoid lines of differentiation.

In one embodiment, the implementation of a tumor or a malignant tumor includes hematopoietic tumor T-lymphoid lines of differentiation.

In one of the embodiments, the tumor is a solid tumor. In one embodiment, the implementation of a tumor or a malignant tumor includes

(a) a tumor with a size of at least about 0.3, 0.4 or 0.5 cm in diameter, or

(b) a tumor that is resistant to treatment with one of at least one of the immunotherapy, angiogenic or chemotherapeutic agent.

In one embodiment, the implementation of support or improve one or more of the number of leukocytes, red blood cells or myeloid cells in the subject.

In one of the embodiments, the tumor reduced in size or substantially eliminated.

In one variant new implementation when an input lactoferrin is administered in a dosage form containing digestible protein, preferably casein or other protein, such as other usable in food proteins.

In one of the embodiments the composition is a food product, beverage, food additive, liquid food Supplement, dietary Supplement, nutritional product, medical food, nutraceutical, medicinal or pharmaceutical. Preferably the composition is for oral or local administration. Preferably the composition is for oral or parenteral administration. In one of the embodiments the composition comprises a milk fat fraction of milk protein.

In one of the embodiments the composition comprises a dairy composition is selected from fresh or whole recombineering milk, recombineering or fresh skim milk, restored whole or skim milk powder, skim milk concentrate, skim milk powder, UltraConnect skim milk, concentrated milk, buttermilk, UltraConnect milk after ultrafiltration, milk protein concentrate (MPC), milk protein isolate (MPI), milk protein concentrate depleted in calcium (MPC), milk with reduced sod is a neigh fat, milk protein concentrate with low fat (MPC), a colostrum fraction, colostrum protein concentrate, colostrum (CPC), whey, colostrum immunoglobulin fraction from colostrum, whey protein isolate whey proteins (WPI), protein concentrate whey (WPC), sweet whey, whey obtained by the use of lactic acid, whey, obtained by using inorganic acids, or restored whey powder.

In one embodiment, the implementation of the milk fat amount for joint introduction of at least one additional drug. In one embodiment, the implementation of the milk fat to be consistent introduction of at least one additional factor.

In one of the embodiments, where the composition according to the invention or the composition used in the method according to the invention, includes lactoferrin composition provides a population of lactoferrin polypeptides or functional variants or fragments where at least approximately 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 99,5 or 100% of the available binding sites of metal ions in the population is associated with a metal ion preferably with the iron ion.

In one of the embodiments, where the composition according to the invention or the composition, skin is fair in the method according to the invention, includes lactoferrin composition provides a population of lactoferrin polypeptides or functional variants or fragments, where approximately 100% of the available population centers of the binding of metal ions associated with a metal ion, preferably an iron ion, and an additional metal ions are associated with molecules of lactoferrin in nonspecific binding sites, thus, lactoferrin at least approximately 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195 or 200% saturated ions metal in stoichiometric ratio.

In one embodiment, the implementation of the milk fat or similar milk fat includes

(a) from approximately 23% (wt./mass.) and approximately 32% (wt./mass.) palmitic acid;

(b) from about 15% (wt./mass.) and about 22% (wt./mass.) oleic acid;

(c) from about 10% (wt./mass.) and about 15% (wt./mass.) stearic acid;

(d) from about 9% (wt./mass.) and approximately 12% (wt./mass.) myristic acid;

(e) from about 3% (wt./mass.) and about 5% (wt./mass.) butyric acid;

(f) any two of a), b), c), d) or e) above;

(g) any three of a), b), c), d) or e) above;

(h) any four of a), b), c), d) or e) above; or

(i) each of a), b), c), d) and e) above.

It should be understood that the indication of some numbers, opican the x in this document (for example, from 1 to 10)also includes an indication of all rational numbers within that range (e.g., 1, 1,1, 2, 3, 3,9, 4, 5, 6, 6,5, 7, 8, 9 and 10), as well as any range of rational numbers within this range (for example, from 2 to 8, from 1.5 to 5.5 and from 3.1 to 4.7) and thus explicitly described all ranges all ranges explicitly described in this document. They are only examples of what is specifically listed, and all of these possible combinations of numerical values between the lowest value and the highest value should be considered as explicitly specified in this application in a similar manner.

In this description, where made reference to patent descriptions, other external documents, or other sources of information, as a rule, this is done in order to provide context for discussing the features of the invention. Unless specifically stated otherwise, reference to such external documents should not be interpreted as recognition that such documents or other sources of information, in any jurisdiction, are prior art or form part of the General prior art in this field.

Also available in a broad sense to say that the invention consists of parts, elements and features referred to or indicated in the description of the application, individually or collectively, in any or all combinations of two and more of these parts, elements or features, and where in this document specify a specific number, which have known equivalents in this area, to which the invention relates, such known equivalents are considered included in the present document, as if they were specified individually.

Brief description of drawings

Figure 1 presents a graph showing that milk fat inhibits the growth of lymphoma and inhibits opukholeobrazovanie. Mice were kept on the control AIN93G diet or the same diet in which the proportion of fat was replaced as milk fat and enriched milk fat. After two weeks of diet in the flanks of mice were injected cell line EL-4 at 2×105. Tumor size was measured by finding two perpendicular diameters (in cm), controlled before 91 days or until the tumor size 1 cm in diameter. Each dot represents the average tumor size with a confidence interval of 95%, as specified 6 mice, and for the indicated number of mice.

Figure 2 presents a graph showing that milk fat enhances the immunogenic treatment with B7-1 eradicate the tumor. Mice were kept on the control AIN93G diet or the same diet in which the proportion of fat was replaced as milk fat and enriched milk fat. After two weeks of this diet in pakamisa was injected 2×10 5cells, EL-4. Controlled tumor size, measured by finding two perpendicular diameters (in cm). When the tumors ~0.4 cm in diameter tumor was injected complexes DNA-liposome containing 60 µg expressing plasmids B7-1. The moment of introduction of the plasmids shown by the arrow. Tumor size was measured by finding two perpendicular diameters (in cm), controlled before 91 days or until the tumor size 1 cm in diameter. Each dot represents the average tumor size with a confidence interval of 95%, as specified 6 mice, and for the indicated number of mice.

Figure 3 presents 4 graph showing that milk fat acts synergistically with saturated iron by lactoferrin (Lf+) in the prevention of tumor formation. (A) Effects on opukholeobrazovanie. Mice were kept on the control AIN93G diet or the same diet, where the proportion of fat and protein was replaced as Lf+, milk fat, and a combination of the Lf+ and milk fat. Day 0 refers to the day when the mice were placed on a proper diet. After two weeks of diet in the flanks of mice were injected with 2×105cells, EL-4. The tumor size is found by measuring two perpendicular diameters (in cm), controlled before 56 days. Each dot represents the average tumor size with trust is inim interval of 95%, specified as 6 mice, and for the indicated number of mice. (B) Effects on antitumor cytolytic activity. Splenocytes were isolated from mice in (A) at 56 days and analyzed for their cytolytic activity against target cells, EL-4. The percentage of cytotoxicity pending against various ratios of effector cells and target (E:T ratio). Each point indicates the average percentage of cytotoxicity, calculated from 6 mice or a specified number of mice. Error bars indicate 95 percent confidence interval. (C) the Effects on apoptosis of tumor cells. Sections were obtained from tumors in (A) at 56 days and were stained by the indirect way end deoxynucleotidyltransferase end labeling breaks desacetyldiltiazem-digoxigenin (TUNEL) and also by way of fluorescence with annexin V (annexin-V-fluos). The number of apoptotic cells was determined by TUNEL method or fluorescence with annexin V (annexin-V-fluos), were analyzed by 10 randomly selected areas of stained tumor sections, visualized through magnification ×40. Apoptotic index (A/I) is the number of cells in apoptosis (positive in the TUNEL or fluorescence with annexin) × (100/total number of cells). Error bars indicate 95 percent confidence interval. (D) Effects on tumor angiogenesis. Sections were obtained from mice in (A) 56 with the TCA and stained as mAb MEC13,3 against CD31, and against CD105 for visualization of blood vessels or alternatively, one minute to collect tissues in the tail vein injected DiO7 in order to visualize the blood flow. Colored blood vessels believed six mice in six blindly selected random fields.

4 shows six graphs showing that a dry mixture of alpha lipid (Phospholac 600TMand sphingomyelin inhibit tumor growth. Mice were kept on the control AIN93G diet or the same diet, where the proportion of fat and protein was replaced as Lf+ (B), Phospholac 600TM(C), sphingomyelin (D), and a combination of the Lf+ and Phospholac 600TM(E) or a combination of the Lf+ and sphingomyelin (F). The tumor size is found by measuring two perpendicular diameters (in cm), pending against time for each mouse individually. Timeline, pending on the x-axis is not linear.

Figure 5 presents a graph showing that milk fat inhibits tumor growth of primary breast cancer. Mice of Balb/c mice were kept on a diet with milk fat or the corresponding control diet. Tumors formed two weeks later by a subcutaneous injection of 2×104tumor 4T1 cells in the right flank of the mice. Paclitaxel was administered I.P. Pavlova., when the tumors had reached a size of 0.5 cm in diameter. Been watching it grow is the tumor in mice and every three days was measured tumor size. *Significant difference (P<0.05) as compared with mice that were kept on a control diet. #Significant difference (P<0.05) as compared with mice that were kept on a control diet and which was administered paclitaxel.

Figure 6 presents a graph showing that milk fat inhibits tumor growth in breast cancer that metastasize to the lungs, and increases the effect of paclitaxel. Mice figure 5 slew 35 days and separated the light. Estimate the number of metastatic tumors on the surface of the lungs of mice that were kept on a control diet, the mice on the control diet and with the introduction of paclitaxel, on the diet from milk fat and on a diet from milk fat and with the introduction of paclitaxel and expressed by the average number ±SEM. *Significant difference (P<0.05) as compared with mice that were kept on a control diet. #Significant difference (P<0.05) as compared with mice that were kept on a control diet and with the introduction of paclitaxel.

Figure 7 presents a graph showing that milk fat inhibits tumor growth breast cancer that metastasize to the liver, and enhances the effects of paclitaxel. Mice, figure 5, slew 35 days and separated the liver, did the liver slices and stained with hematoxylin/eosin. Counting the number of tumor metastases within the liver of mice, which soderglen control diet, which contained the control diet and with the introduction of paclitaxel and diet from milk fat or a diet with milk fat and with the introduction of paclitaxel and expressed the average ±SEM. *Significant difference (P<0.05) as compared with mice that were kept on a control diet. #Significant difference (P<0.05) as compared with mice that were kept on a control diet and with the introduction of paclitaxel.

On Fig presents two graphs showing that milk fat inhibits tumor angiogenesis. Tumor T mice figure 5 was dissected, made the cut and were stained with mAb against CD31 to identify vascular endothelial cells. In random areas, selected blindly, counted the blood vessels, painted mAb against CD31 to determine the average density of vessels (A) or the average distance to the nearest blood vessel, labeled mAb to CD31, from a point location (B). Error bars represent ±SEM. *Significant difference (P<0.05) as compared with mice that were kept on a control diet. #Significant difference (P<0.05) as compared with mice that were kept on a control diet and with the introduction of paclitaxel.

Figure 9 presents two graphs showing that milk fat protects against damage to the intestine caused by chemotherapy. Mice described in figure 5, was slaughtered at 35 days and the neckline is whether skinny intestine, made the cut and were stained with hematoxylin/eosin. (A) Average length (±SEM) of the villi of the jejunum. (B) Mean activity(±SEM) of γ-GGT. *Significant difference (P<0.05) as compared with mice that were kept on a control diet. #Significant difference (P<0.05) as compared with mice that were kept on a control diet and with the introduction of paclitaxel.

Figure 10 is a graphic showing that milk fat prevents apoptosis of intestinal cells caused by chemotherapy. Skinny intestine, as in Fig.9, were stained by the TUNEL method. Counted the number of apoptotic cells in 10 randomly selected crypts and expressed as the number of apoptotic Taurus on the crypt (mean ±SEM). *Significant difference (P<0.05) as compared with mice that were kept on a control diet. # Significant difference (P<0.05) as compared with mice that were kept on a control diet and with the introduction of paclitaxel.

Figure 11 presents three graphs, showing that milk fat inhibits the body weight loss due to chemotherapy, as described in example 14. (A) Milk fat in the diet insignificant increases body weight in healthy mice. Mice within 4 weeks before chemotherapy contained on the control AIN93G diet or a similar diet, where the proportion of fat was replaced as 70%, 25%and 5% milk fat. At the end of the 4 week period recorded an average mass of those who and mice (corresponding to the shapes, provided at the top above the line). (B, C) Milk fat in the diet prevented the body weight loss due to chemotherapy and accelerated weight gain. After 4 weeks of feeding, as specified above, the mice described in (A), was administered 300 mg/kg of cyclophosphamide. Register the percentage change of body weight 4 (B) and 12 (C) days later and compared with the initial body weight. ***P<0.001 and **P<0,01 compared with the control diet.

On Fig presents a graph showing that milk fat in large doses inhibits the loss due to chemotherapy peripheral WBC, as described in example 15. The number of peripheral WBC were recorded for mice with 11 per day injection of cyclophosphamide and 4, 8 and 12 days later.

On Fig presents two graphs showing that milk fat inhibits the loss of cellular content of the spleen due to chemotherapy and accelerates recovery of cell content of the spleen, as described in example 15. (A) Milk fat prevents loss of cellular content of the spleen. Milk fat has no effect on the cellular content of the spleen in case of feeding the mice for 4 weeks before chemotherapy. Significant inhibition of loss of cellular content of the spleen was achieved in 4 days after chemotherapy through all diets with milk fat, on the 8th day through diet with 7% and 25% milk fat and 12 day through diet with 70% of milk fat compared with the control diet. (B) Milk fat stimulates the formation of splenic colony forming units. Significant stimulation of the formation of splenic colony forming units was reached on the 8th day after chemotherapy through all three diets with milk fat compared with the control diet, although for 12 hours, the situation has reversed, as obtained by feeding with milk fat precursor cells are no longer needed. *Significant difference (P<0,05) against mice, which were kept on a control diet.

On Fig presents three graphs that show that a diet with milk fat reduces symptoms of anemia, as described in example 16. For mice on 11 registered number of RBC, HCT level and the levels of hemoglobin in cardiac samples on the day of injection of cyclophosphamide and 4, 8 and 12 days later. (A) diet from milk fat increases the number of RBC, but the results are not significant. (B) Diet from milk fat increases the level of HCT (RBC volume). Two higher doses of milk fat was significantly increased HCT levels at 8 and 12 hours. (C) Diet from milk fat increases the levels of hemoglobin. Two higher doses of milk fat was significantly increased levels of hemoglobin for 12 hours, and diet with 25% milk fat also increased the levels of hemoglobin for 8 days.

On Fig presents a graph showing that a diet with milk fat pre is appalled by the damage of the small intestine, as described in example 17. For mice on 11 registered length of the villi of the small intestine in the day of the introduction of cyclophosphamide and 4, 8 and 12 days later.

On Fig presents a graph showing the recovery of the intestinal villi after the defeat of the small intestine, mediated by cyclophosphamide. Mice were kept on one of four diets (control diet and diets with substitution by 0.025% Lf+, milk fat, and a combination of these) and record the length of the villi of the jejunum in the day of the introduction of cyclophosphamide and 4, 8 and 12 days later. For 8 days the average length of the fibers in the case of the combination of milk fat and 0.025% Lf+ was significantly greater than in the case only of milk fat or only 0.025% of Lf+, providing evidence of a synergistic effect.

On Fig presents a graph showing that after chemotherapy cyclophosphamide in mice, which were kept on a diet containing milk fat, 0,025% Lf+, or a combination of the latter, the number of WBC recovered faster than in mice, which were kept on a control diet. Registered number of WBC in samples of heart blood in mice on Fig daily injection of cyclophosphamide and 4, 8 and 12 days later. An increase in WBC between 4 days and 8 days in the case of the combination of milk fat and 0.025% Lf+ was significantly greater than in the case only of milk fat or only 0.025% of Lf+, both the providing thus evidence of a synergistic effect.

On Fig presents a graph showing that after chemotherapy cyclophosphamide in mice, which were kept on a diet containing milk fat, of 0.25% Lf+, or a combination of the latter, the number of RBC recovered faster than in mice, which were kept on a control diet. In samples of heart blood in mice on Fig recorded the number of RBC per day injection of cyclophosphamide and 4, 8 and 12 days later. For 12 hours the number of RBC for the combination of milk fat and 0.25% Lf+ was significantly higher than for milk fat or only 0.25% Lf+that provided evidence of a synergistic effect.

On Fig presents a graph showing that after chemotherapy cyclophosphamide in mice, which were kept on the diet, including milk fat, of 0.25% Lf+, or a combination of the latter, HCT is recovering faster than mice that were kept on a control diet. HCT (RBC volume) in samples of heart blood in mice on Fig recorded daily injection of cyclophosphamide and 4, 8 and 12 days later. On the 12 days of HCT for the combination of milk fat and 0.25% Lf+ was significantly higher than for milk fat or only 0.25% Lf+, thus providing evidence for a synergistic effect.

On Fig presents a graph showing that after chemotherapy cyclophosphamide in mice, which were kept on the diet, including milk fat ,25% Lf+, or a combination of the latter, hemoglobin is recovering faster than mice that were kept on a control diet. The hemoglobin level was detected in samples of heart blood in mice on Fig the day of the introduction of cyclophosphamide and 4, 8 and 12 days later. On the 12 days of hemoglobin for the combination of milk fat and 0.25% Lf+ was higher than for milk fat or only 0.25% Lf+. The comparison only with 0.25% Lf+ was significant.

On Fig presents a graph showing that milk fat increases body weight after chemotherapy cyclophosphamide in mice, which were kept on a diet of milk fat with Lf+ or without Lf+. Weight was registered tel mice on Fig daily injection of cyclophosphamide and 4, 8 and 12 days later. When data from all day combined, the mass of any mice that were kept on a diet enriched milk fat was significantly higher than in groups, which is not contained in the diet, enriched with milk fat.

Detailed description of the invention

1. Definition

The term "anhydrous milk fat" and "AMF" are used herein interchangeably and refer to fractions of milk fat obtained by contacting phases of cream or melted butter. Milk fat may be any milk fat mammals, including as non-limiting examples of milk fat cow, sheep, goat, pig, mouse, Asian BU the ox, camel, Yak, horse, donkey, llama or human milk fat cow, which is the preferred source. Commonly used methods of obtaining AMF disclosed in Bylund (Ed., 1995), incorporated herein in full. Preferred AMF is, as a rule, approximately 60%, approximately 70%, approximately 80%, approximately 90%, approximately 95%, greater than about 95%, about 96%, about 97%, about 98%, about 99%, about 99.5% pure or 100% fat, more preferably AMF with approximately 99% fat, 99.5% fat or more. AMF additionally repeatedly fractionary in hard (H) and soft (S) fractions, the latter optionally, you can fractionate in "soft solid" (SH) and "soft soft" (SS) fraction, the latter can again further fractionate in "soft soft solid" (SSH) and "soft soft soft (SSS) fractions. As will be clear, each fraction differs in composition of fatty acids. Non-limiting illustrative composition of fatty acids for AMF and fractions derived are shown below in tables 1 to 5.

Table 1
Illustrative composition AMF
Component fatty acidAverage (% wt./mass.) Min (% wt./mass.)Max (% wt./mass.)
C4:0 (butyric acid)3,63,34,1
C6:0 (Capraia acid)2,21,92,4
C8:0 (Caprylic acid)1,21,11,4
C10:0 (capric acid)2,62,22,8
C10:1 (2-decanoate)0,30,30,3
C12:0 (lauric acid)2,92,53,2
C12:1 (11-dolezelova acid)0,10,10,1
C13:0 times. (tridecanoate acid-fu.)0,10,10,1
C13:0 (tridecanoate acid)0,1 0,10,1
C14:0 times. (myristic acid again.)0,20,10,2
C14:0 (myristic acid)10,49,510,8
C14:1 (myristoleate acid)0,90,61,0
C15:0 ISO0,40,30,5
C15:0 antiso0,60,50,7
C15:0 (pentadecanoic acid)1,41,11,5
C16:0 times.0,30,20,3
C16:0 (palmitic acid)28,725,430,4
C16:1 (palmitoleic acid)1,91,62,0
C17:0 ISO0,70,60,7
C17:0 antiso0,50,50,5
C17:0 (margaric acid)0,70,60,8
C17:10,30,30,4
C18:0 (stearic acid)11,510,813,6
C18:1 (oleic acid)23,421,826,4
C18:2 (linoleic acid)1,41,31,7
C18:2 conjug.1,31,01,8
C18:30,80,70,9
C20:0 (arachnid acid)0,20,2
C20:10,30,20,3

Table 2
Illustrative composition fraction H
Component fatty acidAverage (% wt./mass.)Min (% wt./mass.)Max (% wt./mass.)
C4:0 (butyric acid)2,01,82,1
C6:0 (Caproic acid)1,31,21,4
C8:0 (Caprylic acid)0,80,80,9
C10:0 (capric acid)2,21,92,4
C10:1 (2-decanoate)0,20,10,2
C12:0 (lauric acid)3,026 3,4
C12:1 (11-dolezelova acid)0,00,00,1
C13:0 times. (tridecanoate acid-fu.)0,10,10,1
C13:0 (tridecanoate acid)0,10,10,1
c14:0 times. (myristic acid again.)0,10,10,2
C14:0 (myristic acid)11,810,712,6
C14:1 (myristoleate acid)0,60,30,7
C15:0 ISO0,40,30,5
C15:0 antiso0,50,40,6
C15:0 (pentadecanoic acid)1,61,2 1,7
C16:0 times.0,30,30,3
C16:0 (palmitic acid)34,831,536,6
C16:1 (palmitoleic acid)1,31,11,6
C17:0 ISO0,80,70,8
C17:0 antiso0,50,50,6
C17:0 (margaric acid)0,90,80,9
C17:10,20,20,3
C18:0 (stearic acid)15,213,919,7
C18:1 (oleic acid)of 17.015,519,8
C18:2 (linoleic acid) 1,31,11,5
C18:2 conjug.0,80,61,1
C18:30,50,40,6
C20:0 (arachnid acid)0,20,20,3
C20:10,20,10,2

Table 3
Illustrative composition SH faction
Component fatty acidaverage (% wt./mass.)min (% wt./mass.)max (% wt./mass.)
c4:0 (butyric acid)4,03,74,3
c6:0 (Caproic acid)2,42,12,6
c8:0 (Caprylic acid)1,2 1,4
c10:0 (capric acid)2,42,22,7
c10:1 (2-decanoate)0,30,20,3
c12:0 (lauric acid)2,52,32,7
c12:1 (11-dolezelova acid)0,10,00,1
c13:0 times. (tridecanoate acid-fu.)0,10,10,1
c13:0 (tridecanoate acid)0,10,10,1
c14:0 times. (myristic acid again.)0,10,10,2
c14:0 (myristic acid)9,89,010,3
c14:1 (myristoleate acid)0,80,5/td> 0,9
c15:0 ISO0,40,30,4
c15:0 antiso0,50,40,6
c15:0 (pentadecanoic acid)1,41,11,5
c16:0 times.0,20,20,3
c16:0 (palmitic acid)32,829,834,0
c16:1 (palmitoleic acid)1,51,31,8
c17:0 ISO0,60,60,7
c17:0 antiso0,40,40,5
c17:0 (margaric acid)0,80,80,9

c17:10,30,20,3
c18:0 (stearic acid)13,212,516,1
c18:1 (oleic acid)19,517,422,2
c18:2 (linoleic acid)1,31,21,5
c18:2 conjug.1,21,01,6
c18:30,70,60,7
c20:0 (arachnid acid)0,20,20,3
c20:10,20,20,3

td align="center"> 2,0
Table 4
Illustrative composition faction SSH
Component fatty acidThe medium is (% wt./mass.) Min (% wt./mass.)Max (% wt./mass.)
c4:0 (butyric acid)4,0a 3.94,3
c6:0 (Caproic acid)2,42,22,6
c8:0 (Caprylic acid)1,41,21,6
c10:0 (capric acid)2,82,43,4
c10:1 (2-decanoate)0,30,30,3
c12:0 (lauric acid)3,22,7the 3.8
c12:1 (11-dolezelova acid)0,10,10,1
c13:0 times. (tridecanoate acid-fu.)0,10,10,1
c13:0 (tridecanoate acid) 0,10,10,1
c14:0 times. (myristic acid again.)0,20,10,2
c14:0 (myristic acid)11,510,612,2
c14:1 (myristoleate acid)0,90,71,0
c15:0 ISO0,40,40,5
c15:0 antiso0,60,60,7
c15:0 (pentadecanoic acid)1,41,21,5
c16:0 times.0,30,20,3
c16:0 (palmitic acid)28,625,730,0
c16:1 (palmitoleic acid)1,81,6
c17:0 ISO0,70,60,7
c17:0 antiso0,50,50,5
c17:0 (margaric acid)0,70,60,8
C17:10,30,30,4
c18:0 (stearic acid)10,610,211,3
c18:1 (oleic acid)22,220,324,8
c18:2 (linoleic acid)1,41,31,5
c18:2 conjug.1,31,11,7
c18:30,80,81,0
c20:0 (arachnid acid)0,2 0,10,2
c20:10,20,00,3

Table 5
Illustrative composition faction SSS
Component fatty acidAverage (% wt./mass.)Min (% wt./mass.)Max (% wt./mass.)
c4:0 (butyric acid)4,44,0the 4.7
c6:0 (Caproic acid)2,72,42,8
c8:0 (Caprylic acid)1,61,41,8
c10:0 (capric acid)3,42,83,7
c10:1 (2-decanoate)0,40,30,4
c12:0 (lauric acid)3,74,1
c12:1 (11-dolezelova acid)0,10,10,1
c13:0 times. (tridecanoate acid-fu.)0,20,10,2
c13:0 (tridecanoate acid)0,10,10,1
c14:0 times. (myristic acid again.)0,20,20,2
c14:0 (myristic acid)10,29,511,0
c14:1 (myristoleate acid)1,20,81,3
c15:0 ISO0,50,40,5
c15:0 antiso0,80,70,9
c15:0 (pentadecanoic acid)1,10,9c16:0 times.0,30,20,3
c16:0 (palmitic acid)20,018,421,1
c16:1 (palmitoleic acid)2,62,23,0
c17:0 ISO0,60,50,6
c17:0 antiso0,50,50,5
c17:0 (margaric acid)0,40,40,5
c17:10,50,50,6
c18:0 (stearic acid)6,75,87,8
c18:1 (oleic acid)30,828,2the 33.4
c18:2 (linoleic acid) 1,91,72,1
c18:2 conjug.1,71,32,3
c18:31,31,11,4
c20:0 (arachnid acid)0,10,10,4
c20:10,30,10,4

The terms "anticancer factor food product", "anticancer food product" and "anticancer component food product" refers to food products and food components of the product, which is capable of inhibiting tumour formation or growth, and preferably is able to enhance the ability of dairy fat and/or lactoferrin to inhibit the formation and growth of tumors.

The term "anti-tumor factors" refers at least to the factors that cause apoptosis, and may include antitumor cytolytic antibodies and destroy tumor cells cytokines, such as TNF-α.

The term "antitumor immune response" refers to the ability of milk fat or varnish is operina to encourage the formation of antigen specific cytolytic activity (activity of immune cells, particularly cytotoxic T-lymphocytes) and/or activity of NK cells, enhance the cellular immune response to antigens (through the activity of at least cytotoxic T-lymphocytes), improve immune protection (by at least restoring the activity of cytotoxic T-lymphocytes and/or NK cells and increased production of cytokines), restore the immune protection (by at least restoring or stimulating the activity of cytotoxic T-lymphocytes and/or activity of NK cells and increased production of cytokines), the formation of Pro-inflammatory and immunoregulatory mediators (cytokines (Th1 and Th2) and/or education antitumor cytolytic antibodies and destroying tumor cells cytokines, such as TNF-α.

The term "protivokahekticalkie tool" and its grammatical variant protivokahekticalkie facility refers to a facility that can pay, to slow down or stop cachexia or possessing activity to ameliorate one or more symptoms of cachexia, including progressive loss of body weight (including weight loss due to lipolysis and weight loss due to MOLISA), anemia, edema, and anorexia in the subject. Protivokahekticalkie tools include inhibitors of cyclooxygenase (e.g., indomethacin), corticosteroids and glucocorticoids, such as prednisolone, IU ylprednisolone and dexamethasone, progestogenic tools, such as megestrol acetate, medroxyprogesterone acetate, cannabinoids, such as tetrahydrocannabinols and dronabinol, serotonin antagonists such as cyproheptadine at tools that affect intestinal motility, such as metoclopramide and cisapride, steroidal anabolic agent, such as nandrolone decanoate and fluoxymesterone, inhibitors phosphoenolpyruvate-carboxykinase, such as hydrazine sulfate, analogues of methylxanthines, such as pentoxifylline and lisofylline, thalidomide, cytokines and anticytokines, such as antibody Anti-IL-6, IL-12, branched chain amino acids, agents, improving lipid metabolism, such as eicosapentaenoic acid, inhibitors of prostaglandin synthesis, such as indomethacin and ibuprofen, hormones such as melatonin agonists β2-adrenergic receptors, such as clenbuterol, metoclopramide, growth hormone, IGF-1 and antibodies to factors TNF-alpha, LIF, IL-6 and oncostatin M, causing cachexia.

The term "protivopokazanie tool" means a tool that can lead to improvement in the intestine damage, such as alteration or circulation, slow down or stop mucositis or having the ability to ameliorate the symptoms of mucositis.

The term "comprising"as used in this description, means "contains at m is partially re". When interpreting statements in this description, which include that term in each statement should be presented all the characteristic features that begin with this term, but can be also presents other features. Related terms such as "include" and "includes"should be interpreted in the same way.

"Effective amount" is an amount necessary to ensure therapeutic effect. Compliance of dosages for animals and humans (based on milligrams per square meter of body surface) is described by Freireich, et al. (1966). The surface area may be approximately determined from the height and weight of the subject. See, for example, Scientific Tables, Geigy Pharmaceuticals, Ardley, New York, 1970, 537. Effective doses will also change in accordance with the decision of the experts in this field, depending on the method of administration used excipient etc.

The terms "enhance the immune system and stimulate the immune system" (and grammatical times these terms) refer to the ability of milk fat to stimulate the formation of antigen specific cytolytic activity (activity of immune cells, particularly cytotoxic T-lymphocytes) and/or activity of NK cells, enhance the cellular immune response to antigens (through activity measures at the cytotoxic T-lymphocytes), to improve the immune protection (by at least restoring the activity of cytotoxic T-lymphocytes and/or NK cells and increased production of cytokines), restore the immune protection (by at least restoring or stimulating the activity of cytotoxic T-lymphocytes and/or activity of NK cells and enhancing cytokine production) or to form a Pro-inflammatory and immunoregulatory mediators (cytokines (Th1 and Th2).

The term "functional fragment of lactoferrin" is intended to refer to naturally occurring or navstrechalas in the nature part of the lactoferrin polypeptide that has activity, was investigated according to the examples below, and includes functional fragments containing the metal ion. Suitable fragments of lactoferrin include truncated polypeptides lactoferrin, lactoferrin hydrolysates, binding metal ions, fragments that comprise the Central metal ion binding N-lobe, fragments that comprise a binding site metal ion in the C-lobe, and fragments that bind metal ions formed (by artificial or natural processes) and identified by known methods, which are discussed below. In the published international patent applications WO 2006/054908 and WO2007/043900 report on receipt and use of fragments of lactoferrin and they are in the turned off in the present document by reference.

The term "functional lactoferrin variant" is intended to denote a variant of lactoferrin polypeptide that has activity, was investigated according to the examples below, and includes functional variants containing the metal ion.

The term "glycosylated", used in relation to the lactoferrin polypeptide, functional variant or functional fragment is intended to indicate that lactoferrin is fully or partially glycosylated glycosylase groups occurring in nature or newstrategies in the nature of a person or of a bull. Glycosylated and deglycosylated forms of lactoferrin is known (see Pierce, et al. (1991); Metz-Boutigue et al. (1984); van Veen, et al. (2004)).

The term "hematopoietic tool" means a tool that is able to regulate and preferably to stimulate haematopoiesis and/or lymphopoiesis and includes tools that improve the quality of the blood by increasing the number of blood cells such as erythrocytes, lymphocytes or myeloid cells, and by increasing the level of hemoglobin. Preferred hematopoietic tools are suitable in the treatment of anemia. Illustrative hematopoietic tools include hematopoietic, lymphopoiesis and myeloid growth factors and recombinant equivalents, such as the hands of Eritrean who epoetin, including epoetin Alfa, thrombopoietin, IL-1-12, IL-20, a factor that stimulates the formation of colonies of granulocytes/macrophages (GM-CSF), including sargramostim (LEUKINE), the factor stimulating the formation of colonies of monocytes/macrophages (M-CSF or CSF-1), the factor stimulating the formation of colonies of macrophages (M-CSF), factor, stimulating the formation of colonies of granulocytes (G-CSF), including filgrastim (NEUPOGEN), stem cell factor (SCF), the ligand FTL-3 (FL), iron and salt iron, such as ferrous sulfate, ferrous fumarate, gluconate, iron, etc., edetate iron, iron dextran, sodium zelenogradtsy complex (FERRLECIT), pyridoxine, Riboflavin, vitamins, including B12 and folic acid. Cm. also Goodman & Gilman's Pharmacological Basis of Therapeutics, 10th Edition, Harman JG and Limbird LE eds., McGraw-Hill, New York, Chapter 54. Hematopoietic Agents: Growth Factors, Minerals, and Vitamins, pp.1487-1517.

The term "improving the response of the subject" is intended to indicate that the subject has observed a greater reduction in the rate of tumor growth, tumor size, or clinical symptoms than the subject which is not subjected to the method according to the invention. In one of the embodiments being treated, the subject also has the benefit of one or more recovered vitamin status, reduced time of chemotherapy, low dose chemotherapy, increased immune resistance, improve health, is due to the power, reduced cachexia, reduced mucositis, reduced anemia, reduced hematologic suppression or increased haematopoiesis.

The term "increased sensitivity of the tumor" is intended to indicate that the tumors observed a greater decrease in the rate of tumor growth, tumor size, or it is destroyed, at the same time have a tumor that is not influenced by the method according to the invention will not experience these effects.

The term "immunotherapy agent" is intended to refer to an agent that stimulates antitumor immunological activity, also referred to herein as anti-tumor immunity and antitumor immune response(s). Tools that stimulate antitumor immune activity, are preferably those which directly or indirectly stimulate T-cells and/or NK cells to destroy tumor cells. Illustrative analysis to determine stimulates whether the selected tool antitumor immunological activity in vitro, is an analysis of CTL, described below.

The term "inhibition of cancer" is intended to indicate that the tumors are not formed or that tumors are formed, but are not defined or does not grow, or that tumors are formed, but still not the most, benign and do not become cancerous tumors or not metastasize, or that tumors grow more slowly. The tumors can be controlled by images obtained with CT and tumor markers, where available.

The term "inhibition of tumor growth" is intended to indicate that the tumors are not formed in a subject undergoing treatment according to the invention, or one or more tumors that may be present in a subject undergoing treatment according to the invention, does not grow in size or become malignant tumors or not metastasize, or that one or more tumors present in the subject treated according to the invention, decrease in size (preferably, at least about 20, 30, 40, 50, 60, 70, 80, 90 or 100% volume), or one or more tumors present in the subject treated according to the invention, destroyed. Tumor size can be controlled by images obtained with CT and tumor markers, where available.

The term "lactoferrin containing iron and lactoferrin saturated with iron", as used herein, is intended to naming the population of lactoferrin polypeptides, providing its population centers, where at m is re approximately 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 99,5, 99,9 or 100% are represented in the population centers, linking metal ions are bound iron ion.

The term "lactoferrin polypeptide" refers to deglycosylation or glycated amino acid sequence of lactoferrin wild-type or homologous sequences lactoferrin other species, such as described below. The lactoferrin polypeptide has two binding metal ions and thus can bind metal ions in a stoichiometric ratio of 2 metal ion molecule of lactoferrin. One binding metal ions present in the N-terminal lobe (N-lobe) lactoferrin, and the other center is located in the C-terminal lobe (C-lobe) (Moore et al, 1997). Proven sequence of lactotransferrin (predecessors lactoferrin) of bovine and human, lactoferrin and peptides thereof can be found in the database Swiss-Prot (http://au.expasy.org/cgi-bin/sprot-search-ful). These lactoferrin polypeptides include predecessor lactotransferrin bull with registration number P24627, bovine lactoferrin, predecessor lactotransferrin of man with registration number P02788 and human lactoferrin. In the published international patent applications WO 2006/054908 and WO 2007/043900 report on receipt and use of lactoferrin polypeptides and his aminoxy the pilot sequence, and each application incorporated herein by reference. Polypeptides lactoferrin can bind metal ions, as a rule, iron ions, to "natural" levels. For example, in the nature of bovine lactoferrin iron-replete from approximately 10% to 20% (preferably 15%). In the framework of the invention suitable apolactoferrin and lactoferrin at least 1% of saturation of the metal ion.

The term "solid tumor" is intended to refer to tumors that cannot be treated by one of at least one of the immunotherapy, anti-angiogenic or chemotherapeutic agent, preferably not treatable at least one at least one of the immunotherapy or chemotherapy agent. In one embodiment, the implementation of the solid tumor is a tumor that represents at least approximately 0,3, 0,4, 0,5, 0,6, 0.7 or 0.8 cm in diameter. In one embodiment, the implementation of the solid tumor is a tumor that represents from about 0.3 to about 0.8, from about 0.4 to about 0.8, from about 0.5 to about 0.8, from about 0.6 to about 0.8, or from about 0.7 to about 0.8 cm in diameter. In one embodiment, the implementation of the solid tumor is a tumor that does not supports the treatment is immunotherapy or antiangiogenic therapy or chemotherapy.

The term "binding metal ions" is intended to denote the binding of a metal ion in the binding of iron lactoferrin polypeptide or the binding of iron polypeptide fragment of lactoferrin, which is still capable of forming binding of iron.

The terms "containing a metal ion lactoferrin" and "intense metal ion lactoferrin is intended for the naming of the population of lactoferrin polypeptides that provide the population centers of the binding of metal ions where at least about 25% of the binding of metal ions present in the population, have an associated metal ion. It should be understood that the population may contain polypeptides of different types; for example, some molecules that do not bind any one ion, and others, where each connects one or two ions. If used in a variety of metal ions, some molecules can bind a metal ion selected from, for example, the group comprising aluminum, bismuth, copper, chromium, cobalt, gold, iron, manganese, osmium, platinum, ruthenium, zinc ions, or other ions that can in a certain way to set the correct value in the binding of a metal ion in the lactoferrin, and other molecules that can bind a variety of ions. In some cases, the population may include polypeptid is s, involved in the nonspecific binding of ions, where one or more ions, preferably metal ions, non-related polypeptide, i.e. are not in the binding of metal ions. Non-limiting examples of ions that can be associated with nonspecific lactoferrin polypeptides, are calcium and selenium.

The terms containing metal ion fragment of lactoferrin and saturated metal ion fragment of lactoferrin equally intended for naming populations of polypeptide fragments of lactoferrin, which provide binding sites of metal ions where at least about 25% are present in the population centers of binding metal ions are bound metal ion.

According to the present invention can apply a mixture of lactoferrin polypeptides and fragments of lactoferrin. In this embodiment, the population centers of the binding of metal ions composed of two centres for each of lactoferrin polypeptide and one or two centres for each fragment of lactoferrin, depending on the nature of the fragments.

The degree of saturation can be determined by spectrophotometric analysis (Brock & Arzabe, 1976; Bates et al, 1967; Bates et al, 1973). It should be understood that may involve the exchange of metal ions between lactoferrin polypeptides. In one of the variants of implementation of lactoferrin, rich in iron, can be obtained by the method of Law et al. (1977). In another embodiment, the lactoferrin saturated with iron, you can get way Kawakami et al. (1993). Lactoferrin saturated with metal ions, can be obtained by binding metal ions with places of binding metal ions in lactoferrin, including the binding of a metal ion, such as the binding of Fe and other centers of nonspecific binding in the molecule of lactoferrin or lactoferrin fragment.

In one embodiment, the implementation of at least approximately 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 99,5, 99,9 or 100% of the binding of metal ions present in the population of lactoferrin molecules are bound metal ion, and a suitable range you can choose between any of the above values (for example, from about 25 to about 100%, from about 30 to about 100%, from about 35 to about 100%, from about 40 to about 100%, from about 45 to about 100%, from about 50 to about 100%, from about 55 to about 100%, from about 60 to about 100%, from about 65 to about 100%, from about 70 to about 100%, from about 75 to about 100%, from about 80 to the roughly 100%, from about 85 to about 100%, from about 90 to about 100%, from about 95 to about 100%, and from about 99 to about 100%). In one of the embodiments containing a metal ion lactoferrin is a supersaturated lactoferrin.

The term "containing a metal ion functional fragment of lactoferrin" is intended to refer to naturally occurring or not naturally occurring part of the lactoferrin polypeptide that has one or two binding with metal ions and which has an activity defined in the conditions according to the examples below. Suitable fragments of lactoferrin include a shortened polypeptide lactoferrin, lactoferrin hydrolysates, binding metal ions, fragments that contain a Central metal ion binding N-lobe, the fragments that contain the binding site metal ion in the C-lobe and fragments that bind metal ions formed by artificial or natural means and determined by known methods, as discussed below.

The term "milk fat" includes the lipids and the lipid fraction and the hydrolysates of the lipid fraction mammals. Preferred dairy fats are fats milk, especially milk fat milk cows. Preferred milk fat is in the composition of palmitic acid, oleic acid, stearic acid or myristic acid, one or more, which is present as the most numerous fatty acid(s), preferably palmitic, oleic, stearic and myristic acids are present as the most numerous fatty acids. In particularly preferred embodiments, the implementation of the milk fat is composed of (a) substantially the same percentage by weight of palmitic acid, as well as the normal milk fat cow's milk (from approximately 23% (wt./mass.) and approximately 32% (wt./mass.), typically, approximately 28% (wt./mass.) - see table 1,2, PF Fox and McSweeney PLH eds, Advanced Dairy Chemistry Volume 2 - Lipids, 3rd Ed, Springer NY, NY (2006) ISBN-10:0-387-26364-0); (b) substantially the same percentage by weight of oleic acid, as well as the normal milk fat cow's milk (from about 15% (wt./mass.) and about 22% (wt./mass.), typically, approximately 17% (wt./mass.) - see Fox and McSweeny ibid); (c) substantially the same percentage by weight of stearic acid, as well as the normal milk fat cow's milk (from approximately 10% (wt./mass.) and about 15%(wt./mass.), typically, approximately 12% (wt./mass.) - see Fox and McSweeny ibid); (d) substantially the same percentage by weight of myristic acid, as well as conventional milk W the RA cow's milk (from approximately 9% (wt./mass.) and approximately 12% (wt./mass.), typically, approximately 11% (wt./mass.) - see Fox and McSweeny ibid); (e) substantially the same percentage by weight of butyric acid, as well as the normal milk fat cow's milk (from approximately 3% (wt./mass.) and about 5% (wt./mass.), typically, approximately 4% (wt./mass.) - see Fox and McSweeny ibid); (f) any two of a), b), c), d) or e) above; (g) any three of a), b), c), d) or e) above; (h) any four of a), b), c), d) or e) above; (i) each of a), b), c), d) and e) above. The preferred anhydrous milk fat (AMF), especially AMF composition having essentially the same percentages by weight of palmitic, oleic and stearic acid as the normal milk fat of cow's milk, more preferably essentially the same composition of fatty acids that have normal milk fat cow's milk (see Fox and McSweeny ibid).

The term "similar milk fat" includes any combination of vegetable fats, animal fats, or fats of marine products, which are mixed to provide palmitic acid, oleic acid, stearic acid or myristic acid, one or more present as the most numerous fatty acid(s), preferably palmitic, oleic, stearic and myristic acids are presented as the most numerous fatty acids, thus, similar milk fat and eat in part (a) essentially the same percentages by weight of palmitic acid, as well as conventional milk fat cow's milk (from approximately 23% (wt./mass.) and approximately 32% (wt./mass.), typically, approximately 28% ( wt./mass.) - see Fox and McSweeny ibid); (b) substantially the same percentage by weight of oleic acid, as well as the normal milk fat cow's milk (from about 15% (wt./mass.) and about 22% (wt./mass.), typically, approximately 17% (wt./mass.) - see Fox and McSweeny ibid); (c) substantially the same percentage by weight of stearic acid, as well as the normal milk fat cow's milk (from approximately 10% (wt./mass.) and about 15% (wt./mass.), typically, approximately 12% (wt./mass.) - see Fox and McSweeny ibid); (d) substantially the same percentage by weight of myristic acid, as well as the normal milk fat cow's milk (from approximately 9% (wt./mass.) and approximately 12% (wt./mass.), typically, approximately 11% (wt./mass.) - see Fox and McSweeny ibid); (e) substantially the same percentage by weight of butyric acid, as well as the normal milk fat cow's milk (from approximately 3% (wt./mass.) and about 5% (wt./mass.), typically, approximately 4% (wt./mass.) - see Fox and McSweeny ibid); (f) any two of a), b), c), d) or e) above; (g) any three of a), b), c), d) or e) above; (h) any four of the C (a), b), c), d) or e) above; (i) each of a), b), c), d) and e) above. Suitable fats can include food or cooking oils, including palm, olive, soybean, canola, corn, sunflower, safflower, peanut, grape seed, sesame, hazelnut, almond, cashew, hazelnut, macadamia, pecan, pistachio and walnut and other edible oils, including acai berries, amaranth, apricot, argan barbed, artichoke, avocado, babassu, begenovskogo walnut, black currant seed, borage seed oil, Shorea, bottle gourd, pumpkin Buffalo, carob (algaroba), manaki, coriander seed, evening primrose, camelina sativa seeds, cannabis, seeds, cotton tree, lallemantia, seeds pennica meadow, mustard, seeds of okra (hibiscus seeds, Perilla seeds, pequi, pine, poppy seeds, plum, pumpkin seeds, quinoa, chickpeas Abyssinian, rice, tea (Camellia), rasterops, watermelon seeds and wheat germ oil products from the sea, including fats of shellfish, fish, anchovies, fish of lake Baikal, smoked fish, fish Kaha, carp, eel, Pacific smelt, herring, macruronus, tenualosa, pike fish katla, smoked salmon, mackerel, holostea, carp, pilcharda, notothenia, salmon, sardines, shark, sprat, trout, tuna, smelt and swordfish and a combination of any two or more of them.

The term "oral administration" includes oral, buccal, ante the social and intragastric administration.

The term "parenteral administration" includes, as non-limiting examples of local (including the introduction of any skin, epidermal or mucous surface), subcutaneous, intravenous, intraperitoneal, intramuscular and intratumoral introduction (including any direct injection into the tumor).

The term "pharmaceutically acceptable carrier" is intended to refer to media, including as non-limiting examples excipient, diluent or adjuvant, which you can enter the subject as a component of compositions according to the invention. With the introduction in doses sufficient to Institute an effective amount of milk fat, milk derived fat or component thereof, including, for example, cis-9, trans-11 CLA and TVA or the introduction of a lactoferrin polypeptide or functional variant or functional fragment, preferred carriers do not reduce the activity of the composition and is not toxic. Drugs can be administered orally, nasal or parenteral.

The term "subject" is intended to refer to an animal, preferably a mammal, more preferred mammal is a domestic animal or a person. Preferred animals include cats, dogs and horses.

The term "supersaturated lactoferrin" refers to populations gender the peptides of lactoferrin or a functional fragments, providing a population of binding metal ions, where the metal ions are available in sufficient quantity to saturate the 100% binding, and contains the additional metal ions and associated nonspecific sites linking in a lactoferrin polypeptide or fragment of lactoferrin. In other words, given a stoichiometric excess of metal ions. In the composition according to the invention, comprising supersaturated lactoferrin, presents, preferably, a proprietary metal ions, although it may involve the exchange of the metal ion between the centers of the link between non-specific places and binding between the binding and the nonspecific binding sites. Preferably, supersaturated lactoferrin does not form insoluble aggregates. In one embodiment, the implementation of the supersaturated lactoferrin represents at least 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195 or 200% saturated with metal ions, preferably saturated with iron. Suitable ranges of saturation include from about 25 to about 200%, from about 30 to about 200%, from about 35 to about 200%, from about 40 to about 200%, from about 45 to about 200%, from about 50 to about 200%, from priblisitelno to about 200%, from about 60 to about 200%, from about 65 to about 200%, from about 70 to about 200%, from about 75 to about 200%, from about 80 to about 200%, from about 85 to about 200%, from about 90 to about 200%, from about 95 to about 200% and from approximately 100 to approximately 200% of saturation of the metal ion.

The terms "cure" and its derivatives should be interpreted in their broadest possible sense. The term should not be used to indicate that the subject is treated until total recovery. Thus, the term "treat" widely includes the preservation of the disease entity or symptoms substantially constant, the increase in the rate of recovery of the subject, the improvement and/or prevention of the start of symptoms or enhance the severity of a particular condition or extend the quality of life of the subject. The term "treatment" is also commonly includes support good health in susceptible individuals and the formation of protective forces to prevent disease.

The term "variant" refers to a naturally occurring (allelic variant, for example) or not occurring (artificially created mutant, for example) polypeptide l is ceferina or fragment of lactoferrin, which differ from the primary amino acid sequence of the polypeptide lactoferrin wild-type data types (such as listed below) or its fragment attach, remove or replace one or more amino acids.

As a rule, a variant sequence of the polypeptide has an overall qualitative biological activity, which is determined according to the examples below. Additionally, these variants of the polypeptide sequence can have a sequence identity of at least approximately 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%. Also included in the meaning of the term "variant" are homologues of the polypeptide of lactoferrin. The homologue is, as a rule, the polypeptide of various kinds, but having essentially the same biological function or activity as the corresponding polypeptide described herein.

A preferred variant of the polypeptide preferably has at least about 70, 75, 80, 85, 90, 95 or 99% identity, preferably at least about 90, 95 or 99% identity with the sequence of lactoferrin, described herein, include those sequences that are described in published international patent application WO 2006/054908 and WO 2007/043900, which are included in the real document is t as a reference. Variant fragments preferably has at least about 70, 75, 80, 85, 90, 95 or 99% identity, preferably at least about 90, 95 or 99% identity with a fragment described herein, including sequences, are described in published international patent application WO 2006/054908 and WO 2007/043900. Identity can be determined by comparison of the appropriate amino acid sequence c sequence described herein, such as lactoferrin polypeptide or its fragment, using the BLAST Suite of programs (version 2,2,12; 28 August 2005), which is publicly available at NCBI (ftp://ftp.ncbi.nih.gov/blast/).

Conservative substitutions of one or several amino acids in the sequence of the polypeptide lactoferrin, without significant changes in its biological activity, are also suitable. Specialists in this field there are ways to create substitutions of amino acids that are not manifested phenotypically (see, for example, Bowie et al., (1990)).

The term "vitamin D" refers to one or more compounds of the vitamin D is selected from the group comprising vitamin D1 [luisteren], vitamin D2 [calciferol or ergocalciferol], vitamin D3 [cholecalciferol], vitamin D4 [22-dihydroergosterol] and vitamin D5 [situaltional] and any mixture of any two or more of them. The term "analog of vitamin D" refers to any is the connection, which will bind and activate the vitamin D receptor (VDR). VDR is an intracellular receptor that is activated by the ligand, which acts as a transcription factor and binds to the promoter/enhancer regions of genes sensitive elements to vitamin D (VDREs), including as non-limiting examples of genes that have antiproliferative effects on tumor cells by blocking phase G0/G1 cell cycle and inhibits factors that stimulate growth, such as IGF-1 activates a negative growth regulator, such as transforming growth factor beta, what causes apoptosis of the tumor, inhibition of tumor angiogenesis and inhibition of metastasis. Assays for determining binding of the VDR is known; for example, the immunoassays that measure the level of expression of genes regulated by vitamin D. Thus, appropriate vitamin D analogues for use in the present invention, it is possible to quickly determine without undue experimentation.

2. Milk lipids and fractions of lipids

Milk lipids are comprehensively discussed by Fox and McSweeney (2006) and, therefore, included as a reference. Fraction of milk lipids discussed in Dairy Processing Handbook, 1995 and Illingworth, 2002 and Rombaut et al., 2006 and all inclusive, so as a reference.

Examples of suitable fractions of milk lipids p the invention include cream, butter, anhydrous milk fat (AMF) (usually obtained through the treatment phase of the cream or oil), buttermilk, plasma oils, solid fractions of milk fat, soft fractions of milk fat, sphingomyelinase faction, faction shell globules of milk fat, phospholipid fractions, and complex lipid fractions and their combinations and their hydrolysates.

Multistage fractionation of milk fat can be done by differential crystallization. Fractions of milk fat is heated to the set temperature and crystallized or share on solid (stearin) and liquid (olein) fractions. Multistage fractionation means refaccionaria in the subsequent stage of the product fractionation previous stage.

Other fractionation techniques include contacting phases, transesterification, glycerols, solvent fractionation, supercritical fractionation, fractionation, close to supercritical, distillation, fractionation by centrifugation, crystallization slurry, dry crystallization, fractionation with a modifier (e.g., Soaps or emulsifiers) and combinations of these methods.

The lipids present in the compositions according to the invention can be fully or partially modified, including, for example, glycosylated, salirov is installed, esterified, phosphorylated or hydrolyzed natural way, chemical, enzymatic, or any other known in the field of ways.

Lipid hydrolysates can be obtained using known methods, including as non-limiting examples of hydrolysis in acidic medium, the hydrolysis in an alkaline environment, enzymatic hydrolysis, using, for example, lipase, as described in Fox and McSweeney ((2006), Chapter 15 by Deeth HC and CH Fitz-Gerald) and microbial fermentation. How alkaline hydrolysis involves adding 1% KOH (in ethanol) and heating for 10 minutes. The products of hydrolysis can be neutralized with acetic acid or hydrochloric acid.

The substance of the shell globules of milk fat can be distinguished according to the way acidification Kanno & Dong-Hyun, 1990 and further fractionate complex fractions of lipids and lipoproteins by adding methanol, as described by Kanno et al., 1975. The fraction of phospholipids can be distinguished by extraction of the mixture of lipids with acetone according to the method Purthi et al, 1970. The lipid residue can be further enriched lipid complex by selective extraction of simple lipids pentane.

In one embodiment, the implementation of the milk fat comprises one or more fatty acids selected from butyric acid (C4:0), Caproic acid (C6:0), capryl the howling acid (C8:0), capric acid (C10:0), lauric acid (C12:0), myristic acid (C14:0), palmitic acid (C16:0), palmitoleic acid (C16:1), stearic acid (C18:0), oleic acid (C18:1 cis-9; cis-9-octadecenoate acid), elaidic acid (C18:1 trans-9; trans-9-octadecenoate acid), vaccinology acid (C18:1 trans-11; trans-11-octadecenoate acid), cis-vaccinology acid (C18:1 cis-11; cis-11-octadecenoate acid), arachnid acid (C20:0) and beganovi acid (C22:0), optionally substituted by one or more groups selected from hydroxyl, methyl, ethyl and have various groups and salts, esters and amides, and combinations thereof. The optional substituent(s) may be present in any position throughout the length of the carbon chain. Preferred milk fat consists of fatty acids such as butyric acid, Caproic acid, Caprylic acid, capric acid, lauric acid, myristic acid, palmitic acid, palmitoleic acid, stearic acid, oleic acid and elaidic acid, salts, esters and amides, and combinations thereof. These fatty acids are the main components of cow's milk fat.

In one of the embodiments the composition includes mainly contains or consists of at least approximately 0,1, 0,2, 0,5, 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 0, 95, 99, 99,5, and 99.8 or 99.9% by weight of fresh, recombineering or powder whole milk or milk derived, preferably milk fat and suitable ranges, you can choose between any of the above values (for example, from approximately 0.1 to approximately 50%, from about 0.2 to about 50%, from about 0.5 to about 50%, from about 1 to about 50%, from about 5 to about 50%, from about 10 to about 50%, from about 15 to about 50%, from about 20 to about 50%, from about 25 to about 50%, from about 30 to about 50%, from about 35 to about 50%, from about 40 to about 50%, from about 45 to about 50%, from about 0.1 to about 60%, from about 0.2 to about 60%, from about 0.5 to about 60%, from about 1 to about 60%, from about 5 to about 60%, from about 10 to about 60%, from about 15 to about 60%, from approximately 20 to approximately 60%, from about 25 to about 60%, from about 30 to about 60%, from about 35 to about 60%, from about 40 to about 60%, from about 45 to priblizitelen is 60%, from about 0.1 to about 70%, from about 0.2 to about 70%, from about 0.5 to about 70%, from about 1 to about 70%, from about 5 to about 70%, from about 10 to about 70%, from about 15 to about 70%, from about 20 to about 70%, from about 25 to about 70%, from about 30 to about 70%, from about 35 to about 70%, from about 40 to about 70%, from about 45 to approximately 70%, from about 0.1 to about 80%, from about 0.2 to about 80%, from approximately 0.5 to approximately 80%, from about 1 to about 80%, from about 5 to about 80%, from about 10 to about 80%, from about 15 to about 80%, from about 20 to about 80%, from about 25 to about 80%, from about 30 to about 80%, from about 35 to about 80%, from about 40 to about 80%, from about 45 to about 80%, from about 0.1 to about 90%, from about 0.2 to about 90%, from about 0.5 to about 90%, from about 1 to about 90%, from about 5 to about 90%, from about 10 doprinosilo 90%, from about 15 to about 90%, from about 20 to about 90%, from about 25 to about 90%, from about 30 to about 90%, from about 35 to about 90%, from about 40 to about 90%, from about 45 to about 90%, from about 0.1 to about 99%, from about 0.2 to about 99%, from about 0.5 to about 99%, from about 1 to about 99%, from about 5 to about 99%, from about 10 to approximately 99%, from about 15 to about 99%, from about 20 to about 99%, from about 25 to about 99%, from about 30 to about 99%, from about 35 to about 99%, from about 40 to about 99%, and from about 45 to about 99%).

In one of the embodiments the composition includes at least about 0,001, 0,01, 0,05, 0,1, 0,15, 0,2, 0,3, 0,4, 0,5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18 or 19 grams of fresh, recombineering or powder whole milk or milk derived, preferably milk fat, formula (I) or (II) and suitable ranges, you can choose between any of the above values (for example, from about 0.01 to about 1 gram, from about 0.01 to priblisitelno grams, from about 0.01 to about 19 grams, from about 0.1 to about 1 gram, from about 0.1 to about 10 grams, from about 0.1 to about 19 grams, from about 1 to about 5 grams, from about 1 to about 10 grams, from about 1 to about 19 grams, from about 5 to about 10 grams, and from about 5 to about 19 grams).

In one of the embodiments the composition includes, contains mainly or contains approximately 0,1, 0,2, 0,5, 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 99, 99,5, 99,8 or 99.9% by weight of a lactoferrin polypeptide, a functional lactoferrin variant, a functional fragment of lactoferrin containing a metal ion lactoferrin functional variant containing a metal ion lactoferrin or a functional fragment containing the metal ion lactoferrin or a mixture of any two or more of them and suitable ranges, you can choose between any of the above values (for example, from approximately 0.1 to approximately 50%, from about 0.2 to about 50%, from about 0.5 to about 50%, from about 1 to about 50%, from about 5 to about 50%, from about 10 to about 50%, from approximately 15 to approximately 0%, from about 20 to about 50%, from about 25 to about 50%, from about 30 to about 50%, from about 35 to about 50%, from about 40 to about 50%, from about 45 to about 50%, from about 0.1 to about 60%, from about 0.2 to about 60%, from about 0.5 to about 60%, from about 1 to about 60%, from about 5 to about 60%, from about 10 to about 60%, from about 15 to approximately 60%, from about 20 to about 60%, from about 25 to about 60%, from about 30 to about 60%, from about 35 to about 60%, from about 40 to about 60%, from about 45 to about 60%, from about 0.1 to about 70%, from about 0.2 to about 70%, from about 0.5 to about 70%, from about 1 to about 70%, from about 5 to about 70%, from about 10 to about 70%, from about 15 to about 70%, from about 20 to about 70%, from about 25 to about 70%, from about 30 to about 70%, from about 35 to about 70%, from about 40 to about 70%, from about 45 to the roughly 70%, from about 0.1 to about 80%, from about 0.2 to about 80%, from approximately 0.5 to approximately 80%, from about 1 to about 80%, from about 5 to about 80%, from about 10 to about 80%, from about 15 to about 80%, from about 20 to about 80%, from about 25 to about 80%, from about 30 to about 80%, from about 35 to about 80%, from about 40 to about 80%, from about 45 to approximately 80%, from about 0.1 to about 90%, from about 0.2 to about 90%, from about 0.5 to about 90%, from about 1 to about 90%, from about 5 to about 90%, from about 10 to about 90%, from about 15 to about 90%, from about 20 to about 90%, from about 25 to about 90%, from about 30 to about 90%, from about 35 to about 90%, from about 40 to about 90%, from about 45 to about 90%, from about 0.1 to about 99%, from about 0.2 to about 99%, from about 0.5 to about 99%, from about 1 to about 99%, from about 5 to about 99%, from about 10 doprinosilo 99%, from about 15 to about 99%, from about 20 to about 99%, from about 25 to about 99%, from about 30 to about 99%, from about 35 to about 99%, from about 40 to about 99%, and from about 45 to about 99%).

3. Compositions suitable according to the invention

A composition suitable in the framework of the invention, it is possible to make as a food product, beverage, Supplement to the food product, additive for a beverage, dietary Supplement, nutritional product, medical food, nutraceutical, medical or pharmaceutical. Suitable compositions can obtain a specialist in this field with regard to this area and the idea of this description.

In one of the embodiments the present invention relates to the use of milk fat or similar milk fat, optionally with at least one anticancer agent, preferably lactoferrin, in the manufacture of food, beverage, supplements to the food product, additive to drink, dietary Supplement, nutritional product, medical food, nutraceutical, medical or pharmaceutical. Preferably the composition is for oral or local administration. Preferably the composition is for oral or parenteral the aqueous injection. Preferably the composition is for inhibiting tumor growth, inhibiting tumor metastasis, stimulation of apoptosis, stimulation of apoptosis of tumor cells, treatment or prevention of a malignant tumor, increase the response of the subject or the sensitivity of a tumor to treatment, preservation or improvement of the subject has one or more indicators of the number of white blood cells, red blood cells or myeloid cells, raising the subject of the production of cytokines by Th1 and Th2 in the intestine or tumor, treatment or prevention of anemia, cachexia, mucositis we need in this subject or other applications, as described above. Preferred are milk fat or similar milk fat and at least one additional drug, such as one or more anticancer agents, such as lactoferrin, which is described in this document. Preferably antitumor factor is the one described in this document.

In one of the embodiments the composition is in the form of tablets, caplet, pills, hard or soft capsules or lozenges.

In one of the embodiments the composition is in the form of starch capsules, packaged powder, granules, a suspension, an elixir, a liquid solution, drink or any of the other form, you can add to the food product or beverage, including, for example, water or fruit juice. In one of the embodiments the composition is a product for enteral nutrition, a solid product for enteral feeding or liquid product for enteral feeding.

In one of the embodiments the composition further includes one or more components (such as antioxidants)that prevent or reduce degradation of the composition during storage or after injection.

In one of the embodiments of the composition, suitable within the framework of the invention include any food consumer products that may contain fats, fatty acids or lipids. When the composition contains a protein factor, such as lactoferrin, as at least one additional drug, grocery and consumer goods may contain protein. Examples of suitable food consumer products include bakery products, flour, liquid, confectionery, and reconditioned products from fruits, snacks, cereals, spreads, sauces, sauces, dairy products, including ice cream, yogurt and cheese, drinks, including milk-based drinks and dairy drinks (such as milk drinks, including milk shakes and drinking yogurt), milk is the powder, sports supplements, including supplements based on milk and dairy, additives to the food product, such as bulk protein supplements and products that represent dietary supplements, including dietary supplements in pill form. In this embodiment, the composition is suitable herein can also be infant formula, powder or liquid. Similar views can provide suitable nutraceutical compositions suitable in the framework of the invention.

Compositions suitable in the framework of the invention can optionally include other factors, such as calcium, zinc, magnesium, selenium, vitamin C, vitamin D, vitamin E, vitamin K2, complex carbohydrates, food or cooking oils, including palm, olive, soybean, canola, corn, sunflower, safflower, peanut, grape seed, sesame, hazelnut, almond, cashew, hazelnut, macadamia, pecan, pistachio and walnut and other edible oils, including acai berries, amaranth, apricot, argan barbed, artichoke, avocado, babassu, begenovskogo walnut, black currant seed, borage seed oil, Shorea, bottle gourd, pumpkin Buffalo, carob (algaroba), manaki, coriander seed, evening primrose, flax seed, hemp seed cotton tree, lallemantia, seeds pennica meadow, g is richee, seeds of okra (hibiscus seeds, Perilla seeds, pequi, pine, poppy seeds, plum, pumpkin seeds, quinoa, chickpeas Abyssinian, rice, tea (Camellia), milk Thistle, watermelon seeds or wheat germ, or a combination thereof.

Compositions suitable in the framework of the invention, can be constructed, allowing for the introduction of the subject by any selected method of administration, including as non-limiting examples of oral or parenteral (including local, subcutaneous, intramuscular and intravenous) administration.

Basically, for oral administration, diet (food product, additive to the food product or food additive, for example), nutraceutical or pharmaceutical composition suitable herein may be the specialist in this field, according to the known methods of formulation.

Thus, the pharmaceutical composition is suitable according to the invention, it is possible to make with a suitable pharmaceutically acceptable carrier (including excipients, diluents, auxiliaries and their combinations)are selected according to the intended route of administration and standards in the pharmaceutical field. See, for example, Remington''s Pharmaceutical Sciences, 16th edition, Osol, A. Ed., Mack Publishing Co., 1980.

It should be understood that, although the preferred method of introduction is peroral the first, any route of administration may be suitable for any composition according to the invention, including the introduction of multiple way of introduction, including different ways for different media. Thus, it is also assumed inhalation (breathing in the nose or mouth) and vaginal and rectal administration of any composition of the invention. It is also assumed intramedullary, epidural, intra-articular and intrapleural the introduction of any composition according to the invention. Also expected to maintain milk fat or similar milk fat, optionally at least one additional factor, one way of introduction, followed by a separate, simultaneous or sequential introduction of another means the second method of administration; for example, oral administration of milk fat accompanied by the local introduction of at least one additional drug.

Dosage form, fit within the framework of the invention can be administered orally in the form of a powder, liquid, tablets or capsules. If necessary, a suitable dosage form may contain additional means, including emulsifier, antioxidant, flavoring or coloring tools, or be dissolved in the intestine floor. Suitable soluble in the intestinal coating is known. Restorasi is camping in the intestine coating surrounding the active ingredients and prevent the release of active ingredients in the stomach, but ensure the release after the passage of the dosage form through the stomach. Dosage forms suitable in the framework of the invention, it is possible to apply for immediate, delayed, modified, delayed, pulsed-or controlled release of active ingredients. If necessary, appropriate preparations can contain additional tools, including emulsifier, antioxidant, flavoring or coloring tools.

Capsules can contain any of the standard pharmaceutically acceptable substances such as gelatin or cellulose. Tablets can be in accordance with conventional ways, by pressing a mixture of active ingredients with a solid carrier and a lubricant. Examples of the solid carrier include starch and sugar, bentonite. The active ingredients can also be entered in the form of tablets or capsules with a hard shell containing a binder, such as lactose or mannitol, a conventional filler, and means for tableting. Pharmaceutical compositions can also enter parenteral way. Examples of dosage forms for parenteral administration include solutions of active funds in the water, isotonic saline solution or 5% glucose or other well-known pharmaceutically acceptable excipient. Cyclodextrin is s or other solvent means, well-known well-informed in this field can be used as pharmaceutical excipients for delivery of the drug.

Dosage forms for injection can be in the form of liquid solutions or suspensions. Prior to injection can also get the solution or suspension suitable solid forms in the liquid. Dosage form can also be obtained in the form of an emulsion. Milk fat or similar milk fat and at least one additional factor, if present, can be mixed with carriers such as, for example, water, saline, dextrose, glycerol, ethanol or the like, and combinations thereof.

You can get drugs with a slow release, including milk fat, optionally at least one additional factor. Suitable examples of the drug delayed release include semi-permeable matrices of solid hydrophobic polymers containing milk fat, and at least one additional drug, if present, such as lactoferrin or a functional variant or functional fragment. The matrix can be a profiled products, for example patches or microcapsules. Examples of matrices with a slow release include polyesters, hydrogels (for example, poly(2-hydroxyethyl-methacrylate is) or poly(ethenol)), the polylactide (see US 3773919), copolymers of L-glutamic acid and ethyl-L-glutamate, non-degradable ethylene-vinyl acetate and biodegradable copolymers of lactic acid-glycolic acid such as the LUPRON DEPOT™ (microspheres for injection containing a copolymer of lactic acid-glycolic acid and leuprolide acetate).

Compositions for topical administration, including milk fat or similar milk fat and at least one, if present, the additional drug can be obtained in the form of lotions, creams, ointments, pastes, or balms, using for such applications known to the media.

The present invention also relates to a parenteral dosage form including milk fat, optionally at least one additional drug, and at least one anticancer agent. Preferred is at least one antitumor agent selected from paclitaxel, doxorubicin, epirubicin, fluorouracil, cyclophosphamide, methotrexate, expressing plasmid that encodes costimulator t cells, B7-1, and treatment of dendritic cells. Alternative means are selected from any described herein. Preferred are milk fat or similar milk fat, as described above.

The effectiveness of whom is osili, suitable according to the invention can be assessed in vitro and in vivo. See, for example, the examples below. In summary, in one of the embodiments the composition can be tested on its ability, for example, to inhibit the formation and tumor growth in vitro. The composition can be fed or it can enter the animal (e.g. mouse) and then to evaluate the effects on the size or morphology of the tumor in vivo studies. Based on the results, you can determine the appropriate dose range and route of administration.

Compositions suitable in the framework of the invention can be used individually or in combination with one or more other therapeutic agents. The medicinal product may be a food product, beverage, Supplement to the food product, additive to drink, a component of the food product, the component of the beverage, dietary Supplement, nutritional product, medical food, nutraceutical, medical or pharmaceutical. It is preferable for the drug, effective to alleviate one or more symptoms of the condition associated with malignant tumor or associated with anemia, hematologic suppression, mucositis, or cachexia, or associated with a condition caused by anemia (including macrocytic and microcytic anemia), heme is ideological suppression, mucositis or cachexia. Preferred therapeutic agent includes anticancer dietary factors, immunotherapy, hematopoietic funds protivokahekticalkie funds against mucositis and especially preferred drug is lactoferrin.

In one embodiment, the implementation of the milk fat or similar milk fat optionally at least one additional drug is for introducing separately, simultaneously or sequentially with at least one anticancer agent or anti-tumor treatment described in this document.

In one embodiment, the implementation of the milk fat or similar milk fat optionally at least one additional drug is for joint introduction of at least one anticancer agent or anti-tumor treatment described in this document.

In one embodiment, the implementation of the milk fat or similar milk fat optionally at least one additional drug to be consistent introduction of at least one anticancer agent or anti-tumor treatment described in this document.

In od the ohms of options exercise of milk fat or similar milk fat included or entered as adjuvant antineoplastic or anticancer treatment, in which milk fat or similar milk fat increases or enhances the effects of anticancer agents or anticancer treatment. At least one additional drug can be entered separately.

When used in combination c other drug composition suitable in the framework of the invention and the other drug can be administered simultaneously or sequentially. Simultaneous introduction includes the introduction of a single dosage form that contains all the components, or the introduction almost at the same time different dosage forms. Sequential introduction includes the introduction of various schemes, preferably so that there is overlapping parts of the schema in the periods during which envisages the introduction of a composition suitable in this document, and other medicines.

Suitable means, which can be put together compositions according to the invention include chemotherapeutic agents, immunotherapy, protivokahekticalkie funds funds against mucositis, hematopoietic agents and other suitable means known in this field. Preferably such means are administered parenterally, preferably by intravenous, subcutaneous, nutrim the muscle, intraperitoneal, intramedullary, epidural, intradermal, transdermal (topical), transmucosal, intra-articular and intrapleural, as well as oral, inhalation, vaginal, and rectal administration.

It is additionally assumed that the composition in accordance with the invention can comprise additional active ingredients, which can be advantageous to the subject in separate cases. For example, you can use a therapeutic agent having the same target or different aspects of the pathological process.

Suitable means, which can together be entered compositions suitable herein include antagonists integranova receptor alpha v beta 3, antiestrogens or SERMs (selective modulators of estrogen receptor) (including as non-limiting examples of tamoxifen, raloxifene, lasofoxifene, toremifene, atrktive, clomiphene, droloxifene, idoxifene, levormeloxifene, zuclomiphene, enclomiphene, nafoxidine and their salts), protivostoyanie tools, bisphosphonates (including as non-limiting examples of alendronate, clodronate, etidronate, ibandronate, encadrant, minodronate, meridional, olpadronate, pamidronate, Piedmont, risedronate, tiludronate zoledronate and their pharmaceutically acceptable salts), antagonists ka is elevage receptor, calcium supplements, inhibitors of cathepsin K, dual-action (DABAs) (including as non-limiting examples of strontium ranelate), estrogen and estrogen derivatives (including as non-limiting examples of 17 beta-estradiol, estrone, conjugated estrogen, a horse estrogen and 17 beta-ethinyl estradiol), flavonoids, folic acid, osteoanabolic tools, osteoprotegerin, progestin and progestin derivatives (including as non-limiting examples norethindrone and medroxyprogesterone acetate), inhibitors using the ATPase antagonists of VEGF preparations, thiazolidinediones, calcitonin, inhibitors of protein kinase, parathyroid hormone (PTH)analogs PTH, recombinant parathyroid hormone, enhances the secretion of growth hormone, the hormone that stimulates growth hormone, insulin-like growth factor, bone morphogenetic protein (BMP)inhibitors of BMP antagonism, derivatives prostaglandin, fibroblast growth factors, vitamin B6, vitamin D, derivatives of vitamin D (including as non-limiting examples of 1,25-dihydroxycholecalciferol), vitamin K, and derivatives of vitamin K, soy isoflavones, calcium salts, fluorides, statins (including as non-limiting examples of lovastatin, simvastatin, pravastatin, fluvastatin, atorvastatin, tseriwastatina, rosuvastatin and pitavastatin) and their combined what I and other appropriate means, known in this field.

In one of the embodiments the composition is suitable in the framework of the invention, includes or introduced simultaneously or sequentially with other components of milk, such as whey protein milk protein fractions of whey milk (including acid or alkaline protein fractions of whey milk or a combination of both), glycomacropeptide, lactoferrin, vitamin D or calcium, or combinations thereof. Suitable compositions containing milk components include such compositions as a food product, a beverage additive to the food product, additive beverage, dietary Supplement, nutritional product, medical food or nutraceutical. These components can also be applied enriched milk fraction.

It should be understood that additional therapeutic agent listed above (based food products, and pharmaceuticals), can also be used in the method according to the invention, where they are administered separately, simultaneously or sequentially with the composition, suitable within the framework of the invention.

As will be clear dose entered compositions, the period of introduction and General mode of administration may differ from the subjects depending on such factors as the severity of symptoms in a subject, the type of disease to be treated, the chosen method introduction the age and floor and/or the underlying disease entity. However, as a General example, the inventors have considered the introduction of a composition suitable in the framework of the invention, which was introduced in an amount of from about 1 mg to about 1000 mg per kg of body weight per day, preferably from about 50 to about 500 mg per kg per day. In one of the embodiments, the inventors have considered the introduction of pharmaceutical compositions suitable in the framework of the invention, from about 0.05 mg to about 250 mg per kg of body weight.

It should be understood that the introduction may include one dose per day or the introduction of a number of individual doses, as needed. It should be understood that the person skilled in the art will be able, without undue experimentation, based on this area and of this description, to determine the effective doses of this state (including the daily dose and schedule of administration).

The present invention also relates to dietary, nutraceutical or oral pharmaceutical composition, comprising, containing mainly or containing milk fat or similar milk fat in combination with lactoferrin, casein, or other protective protein. Preferably the composition contains generally from about 0.1 to 99 wt%. milk fat or similar milk fat and is roughly 0.1 to 99% of the mass. lactoferrin, casein or other protective protein. A more preferred composition containing generally from about 0.5 to 10 wt%. milk fat and from about 10 to 99 wt%. lactoferrin, casein or other protective protein. The most preferred composition contains mainly approximately 1% of the mass. milk fat and about 20% of the mass. lactoferrin, casein or other protective protein. Preferred are milk fat or similar milk fat, as described above.

In one of the embodiments of the composition according to the invention is a milk fraction, preferably a fraction of milk fat. In one embodiment, the implementation of the milk fraction comprises at least about 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95 or 99% by weight of milk fat and suitable ranges you can choose from any of these values (for example, from about 1 to about 99% by weight, from about 5 to about 99% by weight, from about 10 to about 99% by weight, from about 15 to about 99% by weight, from about 20 to about 99% by weight, from about 25 to about 99% by weight, from about 30 to about 99% by weight, from about 35 to about 99% by weight, from about 40 to about 9% by weight, from about 45 to about 99% by weight, from about 50 to about 99% by weight, from about 55 to about 99% by weight, from about 60 to about 99% by weight, from about 65 to about 99% by weight, from about 70 to about 99% by weight, from about 75 to about 99% by weight, from about 80 to about 99% by weight, from about 85 to about 99% by weight, from about 90 to about 99% by weight, or from about 95 to about 99% by weight). Preferably, the composition includes at least one additional drug, the composition may further contain a fraction of milk protein.

In one embodiment, the implementation of the milk fat is entered as fractions of milk fat. Preferred fractions of milk fat include cream, butter, anhydrous milk fat (AMF) (typically obtained by contacting phases cream or oil), buttermilk, plasma oils, solid fractions of milk fat, soft fractions of milk fat fractions sphingomyelin, faction shell globules of milk fat, phospholipid fractions, and complex lipid fraction, and combinations thereof, and hydrolysates and fractions of the hydrolysates, and combinations hydrolysed and/or non-hydrolyzed fractions.

Suppose the equipment phospholipid fractions can be obtained by separation of anhydrous milk fat (such available at Fonterra Co-operative Group Limited, New Zealand) by thin-layer chromatography and/or liquid chromatography. Using similar methods of separation, it is possible to obtain the desired phospholipid fraction. For example, factions, including all three peak sphingomyelin, including phosphatidylcholine and phosphatidylinositol in equal amounts, a small amount of phosphatidylserine and the first of the three peaks sphingomyelin or includes a large number of phosphatidylethanolamine and ceramides and lysophospholipid obtained in working order by liquid chromatography. Hydrolyzed forms of these fractions can be obtained, for example, adding 1% KOH (in ethanol) and stirring and heating for 10 minutes at a pH of 9.5 to 10.0. Preferably, as a rule, hydrolyzed samples neutralized before further use, for example, acetic acid or hydrochloric acid to pH 7.0 and blow dry in a heated enclosure in an atmosphere of nitrogen.

A commercially available phospholipid fractions suitable for use in the present invention. Preferred are the fraction of phospholipids Phospholipid Concentrate PC600™ and the Ganglioside fraction G600™ (both available from Fonterra Co-operative Group Limited, New Zealand), as in non-hydrolyzed form, in hydrolyzed form. And in this case, these fractions can hydrolyze in the manner specified the WMD above.

In one of the embodiments the composition includes mainly contains or consists of approximately 0,1, 0,5, 1, 5, 10, 15, 20, 25, 30, 35, 40, 45 or 50% by weight of fresh, recombineering or powdered whole milk or milk derived, preferably milk fat and suitable ranges you can choose from any of these values (for example, from approximately 0.1 to approximately 50%, from about 0.2 to about 50%, from about 0.5 to about 50%, from about 1 to about 50%from about 5 to about 50%, from about 10 to about 50%, from about 15 to about 50%, from about 20 to about 50%, from about 25 to about 50%, from about 30 to about 50%, from about 35 to about 50%, from about 40 to about 50%, and from about 45 to about 50%). Preferred is a derivative of milk, selected from recombineering, powdered or fresh milk, restored, whole milk powder, concentrated milk, retentate milk, concentrated milk, retentate milk subjected to ultrafiltration, milk protein concentrate (MPC), milk protein isolate (MPI), milk protein concentrate depleted in calcium (MPC), casein, Casey the ATA, milk fat, cream, butter, anhydrous milk fat (AMF), buttermilk, plasma oils, solid fractions of milk fat, soft fractions of milk fat fractions sphingomyelin, fractions shell globules of milk fat, phospholipid fractions, complex lipid fractions, colostrum fraction, colostrum protein concentrate colostrum (CPC), whey, colostrum fractions of immunoglobulins from colostrum, whey protein isolate whey protein (WPI), whey protein concentrate (WPC), sweet whey, whey obtained by the use of lactic acid, whey, obtained with the use of inorganic acids or the recovered powder serum composition obtained from any faction processed milk or colostrum composition obtained from retentate or permeate obtained by ultrafiltration or microfiltration of any faction processed milk or colostrum, or the composition obtained from the faction, not contacting the chromatographic sorbent (slippage), or adsorbed fraction obtained by chromatographic separation of any recycled fraction of milk or colostrum (including as non-limiting examples of ion exchange and gel permeation chromatography), and their combination and their hydrolysates and fractions of the hydrolysates, and combinations hydrolyzed and/or dehydrodiconiferyl.

In one embodiment, the implementation of the method according to the invention includes the introduction of a mixture of milk fat or similar milk fat and lactoferrin or at least one functional variant or functional fragment of lactoferrin, as described below. Thus, in one of the embodiments the composition includes a mixture of milk fat or similar milk fat and lactoferrin or at least one functional variant or functional fragment of lactoferrin. In an alternative embodiment, the composition includes a mixture of milk fat or similar milk fat and at least one functional fragment of lactoferrin.

The composition of the invention comprising lactoferrin or at least one functional variant or functional fragment, you can also enter via parenteral routes of administration, including as non-limiting examples subcutaneous, intravenous, intraperitoneal, intramuscular, and intratumoral administration. Preferably, the lactoferrin introduced parenterally, by injection. Milk fat or similar milk fat can be entered separately and is preferred oral administration. Specialists in this field are able to get suitable drugs for parenteral administration without undue e is sperimentazione.

In one of the embodiments, the daily dose range for an adult weighing 70 kg (by any route of administration) is from about 0.001 to 250 g of milk fat per day, preferably from 0.001 to 100 g, 0.1 to 30 g, 0.1 to 40 g, 0.1 to 50 g, 0.1 to 60 g, 0.1 to 70 g, 0.1 to 80 g, 0.1 to 100 g, 0.1 to 110 g, 0.1 to 120 g, 0.1 to 130 g, 0.1 to 140 g, 0.1 to 150 g, 0.1 to 160 g, 0.1 to 170 g, 0.1 to 180 g, 0.1 to 190 g, 0.1 to 200 g, 0.1 to 210 g, 0.1 to 220 g 0.1 to 230 g, 0.1 to 240 g or 0.1 to 250 g per day, preferably from 10 mg to 1.5 g/kg/day, preferably 50 mg to 500 mg/kg/day. The highest dose is preferred for short-term treatment and prevention, and the lowest dose in long-term treatment and prevention.

4. Polypeptides lactoferrin

Bovine lactoferrin (bLf) is a single-chain glycoprotein that binds iron with a mass of 78 kDa, which is present in cow's milk. This is a natural protein with protective action, usually the secretory to maintain the normal microflora present in the majority of secreted fluids, including milk, colostrum, tears, nasal secretion, saliva, bile, pancreatic juice, mucus of the intestines and the secret genitals. Bovine lactoferrin secrete neutrophils, and it is presented at a high level in the pockets of bacterial infection. It is a multifunctional protein that can morecontribute iron absorption in the intestine, to ensure the growth of cells in the intestine to protect against infection by microbes to control milopotas, to control systemic immune response and prevent the development of malignant tumors (reviewed in Ward et al., 2002; Brock, J H, 2002; Weinburg, E D, 2001; Conneely, O M, 2001; Tomita, et al., 2002 and Tsuda, et al., 2002).

In addition to suitable lactoferrin polypeptides and fragments listed above, examples of the amino acid sequence and mRNA sequence lactoferrin, which are published and are suitable in the methods of the invention include as non-limiting examples of amino acid (registration number NP_002334) and mRNA (registration number NM_002343) sequence of human lactoferrin; the amino acid (registration number NP_851341 and CAA38572) and mRNA (registration number X54801 and NM_180998) sequence of bovine lactoferrin; the amino acid (registration number JC2323, CAA55517 and AAA97958) and mRNA (registration number U53857) sequence goat lactoferrin; the amino acid (registration number CAA09407) and mRNA (registration number AJ010930) sequence lactoferrin horses; amino acid (registration number NP_999527, AAL40161 and AAP70487) and mRNA (registration number NM_214362) sequence lactoferrin pigs; amino acid (registration number NP_032548) and mRNA (registration number NM_008522) the sequence is lactoferrin mice; amino acid (registration number CAA06441) and mRNA (registration number AJ005203) sequence lactoferrin Asian water Buffalo; and amino acid (registration number CAB53387) and mRNA (registration number AJ131674) sequence of camel lactoferrin. According to the invention, these sequences can be used in wild type or in the form of a variant. Encoded by those sequences of polypeptides can be isolated from a natural source, is obtained as a recombinant protein or receive through organic synthesis using known methods.

Methods of obtaining suitable polypeptides and variants known in this area and are discussed below. Suitable recombinant lactoferrin polypeptides and fragments and methods of obtaining them published in the descriptions of U.S. patents US 5571691, US 5571697, US 5571896, US 5766939, US 5849881, US 5849885, US 5861491, US 5919913, US 5955316, US 6066469, US 6080599, US 6100054, US 6111081, US 6228614, US 6277817, US 6333311, US 6455687, US 6569831, US 6635447, US 2005-0064546 and US 2005-0114911.

Suitable variants include variants of bovine lactoferrin bLf-a and bLf-b (Tsuji, et al. (1989); Yoshida, et al. (1991)). Additional suitable variants include glycosylated and deglycosylated forms of lactoferrin (Pierce, et al. (1991); Metz-Boutigue et al. (1984); van Veen, et al. (2004)) and glycosylation mutants (with option centers glycosylation or option glycosidic residue side chains.

Suitable fragments include fragments of the N-lobe and C-lobe (Baker, et al., 2002) and any other lactoferrin polypeptides that retain the binding of lactoferrin, such as truncated polypeptides lactoferrin. Other fragments of lactoferrin described in published international patent application WO2007/043900, which is incorporated herein by reference.

Suitable truncated lactoferrin polypeptides include polypeptides, shortened from about 1 to about 300 amino acids, preferably approximately 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, 200, 205, 210, 215, 220, 225, 230, 235, 240, 245, 250, 255, 260, 265, 270, 275, 280, 285, 290, 295 or 300 amino acids or more, and include polypeptides, shortened from the N-Terminus, C-Terminus, or with a N - Terminus and C-Terminus, while maintaining in a truncated polypeptide of at least one binding metal ions to the N-lobe or C-lobe. Reported that ligands of metal ions in the N-lobe are residues of amino acids Asp 60, Tyr 92, Tyr 192 and His 253 bovine lactoferrin (without signal sequence). Reported that ligands of metal ions in the C-lobe are residues of amino acids Asp 395, Tyr 433, Tyr 526 and His 595 bovine lactoferrin (without signal sequence) (Karthikeyan et al., 1999).

Suitable variants or fragments of lactoferrin PlaysForSure of the present invention can be obtained by the methods including as non-limiting examples of how changes in proteins of the wild type (see Sambrook, et al. (1989) and in another place the discussion of such methods), such as, but not limited to these, site-specific mutagenesis lactoferrin wild-type and expression obtained polynucleotides; how to get expressing fragments of polynucleotide, such as PCR using random or selected primers; how full or partial proteolysis or hydrolysis of wild-type lactoferrin or variant of lactoferrin polypeptides; and methods chemical synthesis of polypeptides. Variants or fragments of lactoferrin can be obtained by expression from recombinant DNA molecules or RNA from lactoferrin or its variants or fragments. Nucleic acid sequences encoding variants or fragments of lactoferrin, can be inserted into a suitable vector for expression in cells, including eukaryotic cells, such as Aspergillus, but not limited to, or bacterial cells such as E. coli, but not limited to them. Variants or fragments of lactoferrin can be obtained using known methods of PCR, including as non-limiting examples of flawed PCR and changes in DNA. Flawed PCR is a method of carrying out PCR conditions is x, when copying DNA polymerase presents with low precision, so that there is a high rate of point mutations along the full-length PCR product (Leung, et al. (1989); Cadwell, et al. (1992)). Rearrangement of DNA refers to the directed homologous recombination in vitro between DNA molecules with different, but vysokomaslichnoy a DNA sequence-based homology sequence generated by random fragmentation of the DNA molecule, with subsequent fixation of the crossover by extension of the primer in the PCR reactions (Stemmer (1994)). Suitable for use in such methods nucleic acid sequence lactoferrin include those listed above or can be obtained by known methods, including, for example, PCR with reverse transcription (RT-PCR) on RNA isolated from tissues. Suitable primers for RT-PCR can be calculated based on the mRNA sequence listed above. On sale available kits for RT-PCR (e.g., sets of Cells-to-cDNA™ from Ambion, USA).

Variants or fragments of lactoferrin can also get well-known synthetic methods (see Kimmerlin, et al., 2005, for example).

Variants that bind metal ions or fragments of lactoferrin, can be obtained by known methods for isolating polypeptides that bind metals, including as non-limiting Ave is Mer metalroofing chromatography, for example. Suitable variants or fragments of lactoferrin may be associated with free or immobilized metal ions, such as Fe, and cleaned in an appropriate manner. For example, suitable variants or fragments can interact with neutral pH with metal ions immobilized by formation of chelate linkages with the matrix for chromatography, including ligands, iminodiethanol acid or Tris(carboxymethyl)Ethylenediamine. Related variants or fragments can be eluted from the carrier matrix and assemble, reducing pH and ionic strength of the used buffer solution. Variants that bind metal ions or fragments, can be obtained by methods described above and below and described in the examples below.

Functional variants, fragments and hydrolysates of lactoferrin can be obtained by choosing variants, fragments and hydrolysates of lactoferrin and evaluating their effectiveness in the methods of the present invention, by applying the methodology set forth in the examples described below.

In one embodiment, the implementation of lactoferrin is any lactoferrin mammals, including as non-limiting examples of a sheep, goat, pig, mouse, Asian water Buffalo, camel, Yak, horse, donkey, llama, bovine or human lactoferrin. The preferred lactoferrin is lactoferrin the bull.

In another embodiment, the lactoferrin is any recombinant lactoferrin mammal, including as non-limiting examples of recombinant lactoferrin sheep, goats, pigs, mice, Asian water Buffalo, camel, Yak, horse, donkey, llama, bull or man. Preferred lactoferrin is a recombinant bovine lactoferrin. Recombinant lactoferrin can be obtained by expression in a cell-free expression systems or in transgenic animals, plants, fungi or bacteria, or other suitable types.

In yet another embodiment, the lactoferrin isolated from milk, preferably sheep, goats, pigs, mice, Asian water Buffalo, camel, Yak, horse, donkey, llama, cow or the milk man. It is preferable lactoferrin isolated from milk by cation exchange chromatography with subsequent ultrafiltration and diafiltration.

5. The allocation of lactoferrin from milk

The following presents an illustrative method of separation of lactoferrin from milk cows. Fresh skim milk (7 l, pH 6.5) passed through a 300 ml column with sorbent S Sepharose Fast Flow equilibrated water purified system milli Q, a flow rate of 5 ml/min and at 4°C. Unbound protein is washed with water 2.5 volume of the layer and associated protein blueraven stupen is that each of 0.1 M to 0.35 M and 1.0 M sodium chloride, about 2.5 volume of the layer. Lactoferrin, eluruumist in the form of a pale pink stripes in 1 M sodium chloride, collected as one fraction and Valitova against water milli Q followed by lyophilization. Dried powder was dissolved in 25 mm sodium phosphate buffer, pH 6.5, and subjected to re-chromatography on S-Sepharose Fast Flow in the gradient of NaCl up to 1 M in buffer as above, and a flow rate of 3 ml/min Fraction containing lactoferrin proper purity, which was determined by electrophoresis in a gel and HPLC with reversed phase, combined cialisbuy and liofilizovane. The final stage of purification of lactoferrin completed by gel filtration on Sephacryl 300 80 mm dicale phosphate, pH of 8.6, containing 0.15 M potassium chloride. Selected fractions are combined, cialisbuy against water milli Q and liofilizovane. The degree of purity with this method of obtaining the shape of lactoferrin saturated with iron, is more than 95%, as indicated by HPLC analysis and the spectral ratio (280 nm/465 nm), equal to ~19 or less.

Other illustrative methods of selection of suitable fractions of milk, including lactoferrin, is presented in US patent No. 5932259 to Kato et al. and US patent No. 5976597 to Takada et al.

6. Ionized metals or unsaturated lactoferrin

The saturation of the iron reaches through time to relax is of 5 mm nitrilotriacetate iron in molar excess of 2:1 (Foley and Bates (1987)) to a 1% solution of purified lactoferrin in 50 mm Tris, pH 7.8, containing 10 mm sodium bicarbonate. Excess nitrilotriacetate iron removed by dialysis against 100 volumes of water milli Q (updated twice) in total for 20 hours at 4°C. Saturated with iron (holo-) lactoferrin can then be lyophilizate. Different degrees of saturation of the iron can be obtained by providing fewer donor metal ions as described in the examples below. Another way to get lactoferrin containing metal ions, published in published international patent application WO 2006/132553, which included, therefore, as a reference. The way to preserve or improve the quality of the composition of lactoferrin with metal ions during storage published in published international patent application WO 2006/096073, which included, therefore, as a reference.

Unsaturated iron (APO) lactoferrin get through dialysis 1% solution of high-purity sample of lactoferrin in the water against 30 volumes of 0.1 M citric acid, pH 2.3, containing 500 mg/l disodium EDTA, for 30 h at 4°C (Masson and heremans kept carrying out renovation (1966)). Citrate and EDTA are then removed by dialysis against 30 volumes of water milli Q (upgraded once) and the resulting colorless solution can be lyophilizate.

The lactoferrin polypeptide may contain iron ion (as found in nature is polypeptid lactoferrin) or other metal ion (for example, ion copper, ion chromium, cobalt ion, manganese ion or zinc ion). For example, lactoferrin obtained from the milk of a cow, you can remove iron and then fill other type of metal ion. For example, in the same way to fill the iron as described above, it is possible to perform the saturation of copper. For saturation of lactoferrin other metal ions can be used the way Ainscough, et al. (1979).

In one embodiment, the implementation of metal ion selected from the group comprising aluminum, bismuth, copper, chromium, cobalt, gold, iron, manganese, osmium, platinum, ruthenium, zinc, or other ions that can in a certain way to establish the right balance in the binding metal ion lactoferrin. The preferred metal ion is an iron ion.

In the composition for use according to the invention, the lactoferrin polypeptide or a fragment of lactoferrin binding metal ions, can be of the same species or different species. For example, every polypeptides or fragments may contain a different number of metal ions or different kinds of metal ions; or the length of a polypeptide can vary, for example, some are full-length polypeptides, and some are fragments, and each of the fragments may represent separate plot full polypeptide. Such a composition can be obtained from nature is the source or mixing of various types of lactoferrin polypeptides. For example, a mixture of lactoferrin polypeptides of different lengths can be obtained by proteinase splitting (full or partial) of a full-sized lactoferrin polypeptides. The degree of splitting can be adjusted according to methods well known in the field, for example, by varying the number of proteases or the time of incubation, and the methods described below. With full splitting receive a mixture of a variety of full-sized fragments of lactoferrin polypeptides; with partial splitting receive a mixture of full-length lactoferrin polypeptides and various fragments.

7. Obtaining fragments of lactoferrin or lactoferrin hydrolysates

Suitable fragments of lactoferrin described in published international patent applications WO 2006/054908 and WO 2007/043900, each of which is incorporated herein in full. Hydrolysates containing suitable functional fragments can be obtained by selecting appropriate enzymes of known specificity of cleavage, such as trypsin or chymotrypsin, and regulation/restriction of proteolysis by pH, temperature, incubation time and the ratio of enzyme to substrate. Processing similar to the selected peptides can be done using specific endopeptidase. As, for example, a bullish lactoferricin can be obtained through the splitting of the texts bovine lactoferrin pepsin at pH 2.0 V for 45 min at 37°C (Facon & Skura, 1996) or at pH 2.5 and 37°C for 4 h, using the enzyme in the form of a 3% (wt./mass. substrate) (Tomita et al., 1994). The peptide can then be obtained in pure form by HPLC with reversed phase (Tomita et al., 1994) or chromatography based on hydrophobic interactions (Tomita et al., 2002).

Alternatively, the peptides of lactoferrin can be obtained by conventional synthetic Fmoc chemistry as described for calycina-1 person (NH2-FFSASCVPGADKGQFPNLCRLCAGTGENKCA-COOH) and the derived peptide lactoferricin (NH2-TKCFQWQRNMRKVRGPPVSCIKR-COOH) at Viejo-Diaz et al., (2003); and bovine peptide lactoferricin (NH2-RRWQWRMKKLG-COOH), as described in Nguyen et al., (2005); and lactoferrin (NH2-WKLLSKAQEKFGKNKSR-COOH) and shorter fragments, as described in van der Kraan et al., (2004).

Basically, to assess the degree of hydrolysis by comparing the molecular masses of the hydrolyzate and control you can use LTO-page (SDS-PAGE). Exclusion chromatography can be used for separation of the hydrolysate of the various factions and to assess the profile of the molecular mass distribution.

In the preferred hydrolytic method bovine lactoferrin was dissolved to 20 mg/ml in 50 mm Tris pH 8.0, 5 mm CaCl2. Added trypsin (Sigma T8642, TPCK treated, Type XII from bovine pancreas, 11700 units/mg protein) in the enzyme-substrate ratio 1:50 wt./mass. and the mixture is incubated at 25°C for 3 hours Reacts the Yu stopped by adding PMSF to a final concentration of 1 mm and watched the degree of cleavage by LTOs-PAG. Split by trypsin sample (4 ml) was subjected to gel filtration on Sephacryl S300 (Amersham GE) (column 90 cm × 2.6 cm) in 50 mm Tris, 0.15 M NaCl with pH 8.0. The appropriate fractions containing higher amounts of fragments of bovine lactoferrin (Legrand et al., 1984), was then subjected to cation exchange chromatography on S-Sepharose fast Flow (Amersham GE) (column 15 cm × 1.6 cm), using sodium phosphate buffer with a pH of 6.5 and a salt gradient to 1 M NaCl. The final separation of the C lobe and N+C fractions was achieved by an additional gel filtration on Sephacryl S300, as described above, but using 10% vol./about. acetic acid as eluent (Mata et al., 1994). Identity cialisovernight (against water milli-Q) and liofilizovannyh fragments was confirmed by LTOs-PAG and sequencing of N-end of the Edman method.

In another way trypsinogen splitting, such as the above, were separated by HPLC with a reversed phase column (Vydac C18, as well as Superti et al., (2001), and gathered up the fragments with a large mass, corresponding to fragments of the C-lobe and the fragments of the N-lobe. The identity was confirmed by MALDI MS.

In one embodiment, the implementation of the hydrolysates suitable herein contain one or more functional fragments.

8. Anticancer dietary factors

Antitumor components of foods discussed in Park et al., 2002 and Kris-Etherton, 2002.

One is from the embodiments antitumor factor of the food product is selected from vitamin D (including vitamin D1 [luisteren], vitamin D2 [calciferol or ergocalciferol], vitamin D3 [cholecalciferol], vitamin D4 [22-dihydroergosterol] and vitamin D5 [situaltional] and vitamin D5 [7-dehydrocholesterol]), analogues of vitamin D (including as non-limiting examples mentioned below), soy protein, one or more components of soybeans (including those selected from the group including but not limited to omega-3 fatty acids from soy, soybean isoflavones (e.g. genistein and/or daidzein) and peptides lunasin (such as described in U.S. patent US 6107287 and US 6544956, which are incorporated herein by reference, and those that have registration numbers AAE49016, AAE49017, AAP62458 and AAP62459)), polyphenols (from green or black tea, for example), lycopene (or tomato juice, for example), wheat bran, flavonoids (or Apple juice, for example), Inositol, resveratrol (or grape juice, for example), propolis, extracts of mushrooms, anthocyanins (or berry juice, for example), almonds, ginseng extract, hydrolyzed casein and their combination.

Examples of compounds of vitamin D, is suitable in the framework of the invention, include as non-limiting examples of calcitriol (1-alpha,25-dihydroxy[1,25(OH)2D3]; 1,25-dihydroxycholecalciferol), 1,25-digidroksiholekaltsiferol, calcifediol (25-hydroxycholecalciferol), 25-hydroxyethylthio, e is localization (and its predecessor ergosterol), cholecalciferol (and its precursor 7-dehydrocholesterol), doxercalciferol, dihydrotachysterol, paricalcitol, seocalcitol [EB 1089; 1(S),3(R)-dihydroxy-20(R)-(5'-ethyl-5'-hydroxylate-1'(E),3'(E)-Dien-1'-yl)-9,10-scoprega-5(Z),7(E),10(19)-triene)], as well as derivatives, analogs, homologues, predecessors and their metabolites.

In one embodiment, the implementation of the antitumor factor of food selected from the group including anticancer foods and antitumor components of food.

In one embodiment, the implementation of functional food products or their derivatives, which possess properties directed against malignant tumors, can be anticancer foods, including fruits, vegetables, legumes, nuts, seeds, grains, spices, herbs, mushrooms, probiotics, apples, apricots, beans (for example, green beans, black beans), chick peas, berries (such as blueberries, raspberries), vegetables from the cruciferous family (e.g., broccoli, Brussels sprouts, cabbage, cauliflower, braunkohle, cabbage leaf, kohlrabi, Chinese cabbage, radish, mustard and turnip), carrots, cheese, corn products, cranberry, eggplant, flax seed, vegetables of the family Alliaceae [e.g., garlic, onion, spring onion (onion), Snet-onion, leek, shallot], ginger (including components of ginger gingerol, PA is Adol and beta-elements), ginseng, grapefruit, grapes, grape juice, green or black tea, ginger, kiwi, sweet potato, leeks, lemons, lime, Noni fruit, onions, oranges, peanuts, peppers, products of rye, salmon, products made from soy milk, soy nuts, soy beans, pumpkins, tangerines, tomatoes, products of wheat bran, rice, papaya, asimina, peaches, persimmon, strawberry, Taro leaves, green bananas, mango, watercress water, yams, almonds, and combinations thereof.

In one embodiment, the implementation of the antitumor component of food you can choose from a group comprising soy protein, one or more components of soybeans (including those selected from the group including but not limited to, omega-3 fatty acids from soy, soybean isoflavones (e.g. genistein and/or daidzein) and peptides lunasin (such as described in U.S. patent US 6107287 and US 6544956, which are incorporated herein by reference, and those that have registration numbers AAE49016, AAE49017, AAP62458 and AAP62459)), cartilage sharks, extract of garlic dietary Supplement with selenium, extract of tea (for example, polyphenols/catechins/epigallocatechin gallate green or black tea), curcuminoid, caffeine, carnosol acid, capsaicin, sesquiterpenes lactones (for example, parthenolide, Kostunica, imagine), Kotilainen A, M. perennials, arginine, glutamine, retinoids of green vegetable leaves, cocoa powder, face of the pin, glucosinolates from cruciferous vegetables, organic sulfur compounds (allicin, diallyl sulfide, diallyl disulfide, allyl mercaptan, N-acetyl cysteine, connections, onion, carotenoids (including as non-limiting examples of (β-carotene), coumarins, dietary fiber, dithiolthione, flavonoids (e.g., myricetin, quercetin, rutin), indoles, Inositol, Inositol hexaphosphate, soy isoflavones (e.g. genistein, daidzein), isothiocyanates, monoterpene (for example, lemon, parilova acid, metol, carveol), wheat bran, diterpene esters, polyphenols, Riboflavin-5'-phosphate, cinnamic aldehyde, vanillin, oksikumarin, phenols (for example, cinnamic acid), polyphenols, plant sterols (e.g., sitostanol, stigmasterol, campesterol), acylglycerol, ticosteroid, protease inhibitors, saponins, isoprenoids, terpenoids, tocotrienols, retinoids, ellagic acid, polyamine, resveratrol, hydroxycotinine acid [e.g., (E)-ferulic acid and (E)-p-coumaric acid], chlorophyllin, propolis and some of its components (e.g., caffeic acid, phenyl complex esters, intellipen C), red wine, tannic acid, extracts of mushrooms, anthocyanins (for example, cyanidin), mushrooms (β-glucan) (e.g., lentinan), extracts of spinach leaves, a mixture of natural antioxidants from the leaves of spinach,Noni juice, vitamins A, B6, C and E, extracts of Cassia Siamese, an extract of beet ordinary, extracts of lemon grass and bamboo sorghum, karnosky acid, capsaicin, sesquiterpenes lactones (for example, parthenolide, Kostanai, Imogen), Kotilainen a, M. perennials and omega-3 fatty acids (including eykozapentaenovuyu acid (EPA) and docosahexaenoic acid (DHA)), and combinations thereof.

In one embodiment, the implementation of anticancer food component selected from the group comprising vitamin D, vitamin B6, taurine, arginine, glutamine, α-lactalbumin, whey colostrum, full or partial hydrolysates of casein, the peptide(s) of casein, known as immunostimulatory (for example, immunocytokines, caseinophosphopeptides, casomorphine, Kaskinen), peptides, colostrum, colostrum, calcium and phosphate of calcium, folate, milk proteins, rich in cysteine, lactoperoxidase, HAMLET (the complex of α-lactalbumin and oleic acid), fragments of plasminogen, prosaposin, saposin, catalase, lactoperoxidase, protein, fatty acid binding, ribonuclease, an inhibitor of beta-glucuronidase, RAZVI, RAZVI, CD36, interferon, tumor necrosis factor, interleukin 2 (IL-2), kininogen and fragments, connotation, cystatin C, fetuin, defensive neutrophils, interleukin 12 (IL-12), chitinozoans proteins, dystroglycan, prostasin, SPARC-like proteins and thrombospondin or the combination.

Protein soy

Soy is already promoted as a tool that helps maintain a healthy heart and healthy bones, prevents the development of malignant tumors and relieves the symptoms of menopause (Kerwin, 2004). Anticancer effects of soy are associated with soy protein which is a protein with a lower content of sulfur-containing amino acids than animal proteins, and has been shown to inhibit the development of tumors in animals, caused by the action of the carcinogen. Other components of the soybean with anticancer activity include protease inhibitors, isoflavones, such as genistein, which may have anti-cancer or pre-cancerous effects and saponins.

Vitamin D and ligands of the receptor for vitamin D

Widely described anti-cancer properties of vitamin D (Harris, et al., 2004). In addition, I believe that vitamin D reduces the risk of the development not only of malignant tumors, but also many common and serious diseases, including type 1 diabetes, multiple sclerosis, cardiovascular disease and osteoporosis (Holick, 2004). Dairy products are the main dietary source of vitamin D in countries such as New Zealand, where milk and other dairy products are fortified with vitamin D. Vitamin D3 is a 25-gidroksilirovanii in the liver and converted in the kidneys and peripheral organs in the active gormonalnomu 1-alpha,25-dihydroxy [1,25(OH)2D3] (calcitriol), which affects many processes associated with growth and development of cells. It has antiproliferative effects on tumor cells by blocking phase G0/G1 of the cell cycle, which leads to the suppression factors that stimulate growth, such as IGF-1, and activates growth regulators with inhibit action, such as transforming growth factor beta. It causes apoptosis of the tumor and is an inhibitor of angiogenesis and tumor metastasis (Nakagawa, et al., 2005). The introduction of the active metabolite 1,25(OH)2D3 at doses required to inhibit the growth of tumors associated with toxicity caused by hyperkalemia. As described, the synthetic structural analogs of 1,25(OH)2D3, which do not cause hyperkalemia, inhibit tumor growth and lead to regression of tumors in animal models (Colston et al., 2003; Nolan, et al., 1998; van Weelden et al., 1998). Anticancer activity of the ligand of the vitamin D receptor (VDR) has been shown in models of carcinoma of the bladder, breast, colon, endometrium, kidney, lung, pancreas, prostate, sarcoma of soft tissue and bone, neuroblastoma, glioma, melanoma, squamous cell carcinoma (SCC) and other (Beer, et al., 2004).

In phase I clinical studies have shown that the dose periodically, once per week, can significantly increase the dose to get the potential therapeutic peak concentrations of calcitriol (Beer, et al., 2001). In the phase II studies have described the levels, contributing to the antitumor activity of the combination of high doses of 1α,25-dihydroxy vitamin D and docetaxel was administered on a weekly scheme subjects with androgen-independent prostate cancer (Beer, et al., 2003).

Several studies have shown that daily intake of 10,000 IU is safe for humans (in the absence of the sun). In most cases, the toxicity of vitamin D, as described, appears after ingestion per day more than 50,000 IU for several years. Some argue that vitamin D is not very toxic (Saul, 2003).

Compounds that act as ligands of the vitamin D receptor (VDR) and which may be suitable in the framework of the invention, generally divided into three classes: (1) "deletenode"that have secosteroids frame structure, (2) "pseudo-secosteroid", which have the A-ring of vitamin D, but one C or D ring is open, and (3) "not-secosteroid", which in its structure is different from that of secosteroids. For an overview of suitable analogues of vitamin D, see Guyton, et al., 2003; Guyton, et al., 2001; Peleg, et al., 2003; Yee, et al., 2005.

Suitable analogues of vitamin D are described in international application PCT/NZ2007/000389 (quoted in it references), incorporated herein in full.

Evaluation of these analogues in the method according to the invention m is should be done by the following protocols described in the examples below.

9. Immune stimulation

The authors of the present invention have found that milk fat, optionally at least one additional drug is able to stimulate and strengthen the immune system. In particular, as shown in the examples below, milk fat, optionally at least one additional factor that can stimulate the formation of antigen specific cytolytic activity (activity of immune cells, particularly cytotoxic T-lymphocytes) and/or the activity of NK cells, enhance the cellular immune response to antigens (through the activity of at least cytotoxic T-lymphocytes), improve immune protection (by at least restoring the activity of cytotoxic T-lymphocytes and/or NK cells and increased production of cytokines), restore the immune protection (by at least restoring or stimulating activity cytotoxic T-lymphocytes and/or activity of NK cells and increased production of cytokines and production of Pro-inflammatory and immunoregulatory mediators (cytokines (Th1 and Th2). I believe that any functional variant of milk fat, including fractions of milk fat and the like, in combination or not in combination with at least one additional drug will show aktivnosti, similar milk fat. Similarly, suppose that when lactoferrin is subject to the introduction, any functional variant or functional fragment of lactoferrin will show activity similar to lactoferrin.

As shown in the examples below, the milk fat is effective for increasing the formation of antigen specific cytolytic activity and/or activity of NK cells, enhance the cellular immune response to antigens, enhance immune protection and restoration of immune defense.

Thus, the present invention relates to a method of stimulating the immune system of a subject, comprising the introduction of milk fat or similar milk fat to a subject, optionally at least one additional drug, such as one or more anticancer agents. The present invention also relates to methods for increasing the production of cytokines by Th1 and Th2 in the tumor of the subject, increase the production of cytokines by Th1 and Th2 in the intestine of a subject, increasing the level of Th1 cytokines and Th2 in the circulatory system of the subject and enhance antitumor immune response in the subject.

In one embodiment, the implementation of these methods according to the invention, the subject is undergoing or will undergo treatment for a malignant tumor, as described above.

In one of the embodiments of subje the t took the treatment, but he had a relapse or susceptible to relapse. In one of the embodiments the subject has a tumor, immune therapy, chemotherapy, antiangiogenic or immunotherapy agent. In one of the embodiments of the subject had previously been unsuccessful treatment of chemotherapy, antiangiogenic or immunotherapy agent.

In one embodiment, the implementation of the Th1 cytokine selected from IL-18, TNF-α and IFN-γ. In one embodiment, the implementation of the Th2 cytokine selected from IL-4, IL-5, IL-6 and IL-10. In one embodiment, the implementation level of Th1 or Th2 cytokines increased at least approximately 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750 or 800%.

If necessary, these methods can be combined with methods of treatment using any one or more anticancer agents (including chemotherapeutic drugs or immunotherapy) or anticancer treatments, described below.

10. Warning malignant tumor

The authors of the present invention have found that milk fat and individually, and in combination with at least one additional drug may inhibit the formation of tumors and inhibit tumor growth and inhibit metastasis of the tumor. Milk fat, an individual is about or in combination with lactoferrin and especially with lactoferrin, containing metal ion, releases or stimulates the release of antitumor factors, such as T-cells and/or NK cells (natural killer cells), and factors causing apoptosis, inside the system of blood circulation, stimulates antitumor immunological activity and manifests immunostimulirutuyu activity, angiogenic activity and direct tumor cytotoxicity and can cause apoptosis of tumor cells, as shown in the examples below. I believe that any functional variant of milk fat, or fractions of milk fat, or similar milk fat will show activity similar to milk fat.

The present invention is useful for the prevention of malignant tumors, especially to prevent relapse (tumor growth) after surgery, which often occurs due to the growth and proliferation of secondary tumors, prevent tumor spread after her diagnosis and the preparation of the subject for the introduction of antineoplastic or anticancer treatment.

Solid tumors must form new blood vessels before will be able to grow beyond a certain size. Thus, inhibition of angiogenesis, especially tumor angiogenesis (formation of blood vessels for the blood supply of the tumor is s) has obvious applications in the treatment of malignant tumors (Dass, 2004). As shown in the examples below, the oral input milk fat, both individually and in combination with lactoferrin and especially with lactoferrin containing a metal ion capable of significantly reducing the number of vessels and significantly reduce the blood flow in tumors.

Inhibition of angiogenesis also finds application in other diseases, including as non-limiting examples of cardiovascular diseases (atherosclerosis and restenosis, for example), chronic inflammation (rheumatoid arthritis and Crohn's disease, for example), diabetes (diabetic retinopathy), psoriasis, endometriosis, macular degeneration and obesity. Therefore, milk fat or similar milk fat, preferably in combination, more preferably at least one additional drug, lactoferrin or a functional variant or functional fragment and particularly those containing metal ion lactoferrin, have application for the treatment and prevention of malignant tumors.

Similarly, oral introduce milk fat or similar milk fat, both individually and in combination at least one additional drug, such as lactoferrin and especially the content is relevant metal ion lactoferrin, able to cause apoptosis of tumor cells, as shown in the examples below. The examples also show that apoptotic factors detected in the serum of mice that contained in the lactoferrin-containing metal ion.

Thus, the present invention also relates to methods of inhibiting cancer in the subject, stimulation of apoptosis in the subject, stimulation of apoptosis of tumor cells in the subject and inhibition of tumor angiogenesis in a subject, comprising an introduction to the subject of lactoferrin or a functional variant containing metal ions or a functional fragment.

The present invention also relates to a method of maintaining or increasing the level of antitumor factors in the blood stream.

In one of the embodiments a subject prone to the formation of malignant tumors. In one of the embodiments the subject has a tumor that is resistant to treatment with chemotherapy, antiangiogenic or immunotherapy agent. In one of the embodiments of the subject had previously been unsuccessful treatment of chemotherapy, antiangiogenic or immunotherapy agent.

If necessary, these methods can be combined with methods of treatment using any one or more anticancer agents including chemotherapeutic drugs or immunotherapy) or anticancer treatments described below.

11. The treatment and prevention of malignant tumors by means of combined treatment

The authors of the present invention have found that milk fat is able to inhibit tumor growth and inhibits tumor metastasis. Milk fat acts synergistically with immunotherapy (including that mediated gene transfer B7-1 inside the tumor), with chemotherapy including paclitaxel, doxorubicin, or epirubicin fluorouracil) or therapy of dendritic cells for the elimination of tumors significantly. Milk fat is able to act synergistically with chemotherapy including paclitaxel, doxorubicin, epirubicin, fluorouracil, cyclophosphamide, or methotrexate)to inhibit the growth and metastasis of tumor. I believe that any functional variant of milk fat, including fractions of milk fat, etc. or similar milk fat, in combination or not in combination with at least one additional drug will show activity similar to milk fat. Similarly, suppose that when lactoferrin is subject to the introduction, any functional variant or functional fragment of lactoferrin will show activity similar to lactoferrin. Milk fat individually or in combination with other treatment modalities was capable of inhibition is th the development of tumors of the line 4T1 breast cancer, which spread to the lungs and liver, and thus, is able to inhibit tumor metastasis.

As described above, it has been found that milk fat releases inside or stimulates the blood flow antitumor factors, such as T-cells and/or NK cells (natural killer cells), and the factors that cause apoptosis, shows immunostimulirutuyu activity, angiogenic activity and direct tumor cytotoxicity and can cause apoptosis of tumor cells, as shown in the examples below.

In one embodiment, the implementation of the chemotherapeutic agent represents paclitaxel, doxorubicin, epirubicin, fluorouracil, cyclophosphamide or methotrexate.

In addition to the methods described above, the present invention relates to a method of inhibiting tumor growth or inhibiting tumor metastasis in a subject and methods of treating or preventing malignant tumors in a subject, where the method includes:

(a) the introduction of milk fat, optionally at least one additional drug and

(b) separate, simultaneous or sequential introduction of at least one antineoplastic or anticancer treatment.

In one of the embodiments the subject suffers from slocate is the result of a tumor or susceptible to the development of malignant tumors. In one of the embodiments, the patient has a tumor, immune therapy, chemotherapy, antiangiogenic or immunotherapy agent. In one of the embodiments, the patient had previously undergone unsuccessful treatment of chemotherapy, antiangiogenic or immunotherapy agent.

In one embodiment, the implementation of at least one anti-tumor agent administered orally or parenterally, although the preferred method of administration depends on the selected anti-cancer drugs. Preferably at least one anti-tumor agent administered orally or by intravenous, intraperitoneally or intratumoral injection. Paclitaxel, doxorubicin, epirubicin, fluorouracil, cyclophosphamide and methotrexate preferably introduced via intravenous or intraperitoneally injection. Expressing plasmid encoding B7-1, preferably introduced via intratumoral injection. Alternatively, from the subject can be obtained tumor cells, transliterate expressing B7-1 plasmid ex vivo, then transfetsirovannyh cells to enter the subject. Alternatively, a soluble fusion protein B7-Ig can be entered parenteral. Therapy dendritic cells are preferably introduced via intravenous, intraper thealloy or intratumoral injection.

In one embodiment, the implementation of the milk fat or similar milk fat administered orally or parenterally.

In one embodiment, the implementation of the milk fat or similar milk fat, optionally at least one additional drug is administered daily for at least about 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 weeks before the introduction of antineoplastic or anticancer treatment.

In one embodiment, the implementation of the milk fat or similar milk fat, optionally at least one additional drug, entered into a period of at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or 21 days, or within at least approximately 1, 2, 3, 4, 5, 6, 7 or 8 weeks, or for at least about 1, 2, 3, 4, 5 or 6 months before the introduction of antineoplastic or anticancer treatment.

In one embodiment, the implementation of the milk fat or similar milk fat, optionally at least one additional drug, entered into a period of at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or 21 days, or within at least approximately 1, 2, 3, 4, 5, 6, 7 or 8 weeks, or for at least about 1, 2, 3, 4, 5, or 6 months after the achala introduction antineoplastic or anticancer treatment.

Preferably milk fat or similar milk fat, optionally at least one additional drug is introduced at least once a day, including continuous oral administration within days, parenteral administration by infusion or by combining methods of administration (oral and parenteral, for example).

In one embodiment, the method of the invention, the tumor is a large tumor, as described above.

In one embodiment, the method of the invention one or more indicators of the number of white blood cells, red blood cells or myeloid cells in the subject are retained or improved.

In one embodiment, the implementation of the milk fat or similar milk fat and at least one additional drug, if entered, is entered in the dosage form, comprising digestible fat, more preferably digestible fat and digestible protein, preferably lactoferrin, casein or other protein, such as other proteins suitable for human consumption.

In one embodiment, the implementation of the milk fat or similar milk fat and at least one additional drug to provide a synergistic therapeutic effect that is greater than the individual effects to the each, preferably greater than the sum of effects of each individually. For example, there is preferably greater effect on inhibition of the formation or tumor growth, regression of tumor cytolytic effects, immune activation, production of cytokines by Th1 and Th2, maintain, or improve the quantity of white blood cells, red blood cells or myeloid cells, treatment or prevention of anemia, cachexia, mucositis, or the response of the subject or of the tumor to treatment method.

These methods can be combined with methods of treatment using any one or more anticancer agents (including chemotherapeutic agents, immunotherapy, protivokahekticalkie funds funds against mucositis or hematopoietic funds) or anticancer treatments, described below.

In one embodiment, the implementation of antitumor treatment selected from the treatments, but not limited to, such as surgery, chemotherapy treatments, methods of radiation therapy, the methods of hormonal therapy, biological methods of treatment/immunotherapy, how antiangiogenic therapy, methods of cytotoxic therapy, vaccines, vaccines based on nucleic acids (e.g., nucleic acids expressing the antigen of a malignant tumor, such as the NC vaccines, including vaccines p185), methods of treatment based on viruses, such as adeno-associated virus, lentivirus), methods of gene therapy, methods of therapy based on low molecular weight inhibitor, methods of therapy on the basis of nucleotides (for example, Rnci, antisense nucleotides, ribozymes etc), methods of treating an antibody-based, oxygen - ozone therapy, treatment by embolization and/or oily chemo-embolization.

In one embodiment, the implementation of anti-tumor treatment or anti-tumor agent selected from chemotherapeutic agents, including as non-limiting examples are listed in the published international patent application WO 2006/054908 fully incorporated herein by reference.

In one of the preferred embodiments chemotherapeutic agent selected from paclitaxel, doxorubicin, epirubicin, fluorouracil, cyclophosphamide and methotrexate.

In one embodiment, the implementation of the anti-cancer agent is an immunotherapy agent. Immunotherapy product is preferably expressing plasmid encoding costimulatory t cells, B7-1, costimulator T-cells or functionally related molecule, such as chimeric molecule B7-Ig.

In one embodiment, the implementation of protivo the Holevo remedy or treatment includes therapy dendritic cells.

In one embodiment, the implementation of the radiation therapy comprises radiation therapy external radiation (including therapy gamma rays and x-rays) and internal radiation therapy using radioactive isotopes. According to the invention, radioactive isotopes can also be used as anticancer agents.

The inventors have shown in the examples below that oral input milk fat, or individually, or in combination with at least one additional drug, such as lactoferrin and especially lactoferrin containing a metal ion capable of increasing the response of the subject and to increase the sensitivity of tumors to anticancer drugs. When lactoferrin is administered in combination with milk fat, believe that any functional variant, functional fragment, functional variant, containing metal ions, or a functional fragment of lactoferrin containing metal ions, will show activity similar to lactoferrin containing metal ion in combination with milk fat.

Thus, the present invention also relates to a method of improving the response of a subject to treatment, such as anti-cancer therapy selected from the group including surgical intervention, methods of chemotherapy, the way the radiation therapy, methods of hormonal therapy (e.g. tamoxifen, aromatase inhibitors), methods of biological treatment/immunotherapy, how antiangiogenic therapy, methods of cytotoxic therapy, vaccines, vaccines based on nucleic acids (e.g., nucleic acids expressing the antigen malignant tumors, such as DNA vaccines, including vaccines p185), treatment-based viruses (e.g., adeno-associated virus, lentivirus), methods of gene therapy, methods of therapy based on low molecular weight inhibitor, methods of therapy on the basis of nucleotides (for example, Rnci, antisense nucleotides, ribozymes etc), methods of treatment based on antibodies, oxygen and ozone therapy, treatment by embolization and/or oily chemo-embolization, or a combination, where the method includes along with treatment of the separate, simultaneous or sequential introduction of milk fat or similar milk fat, optionally at least one additional pharmaceutical agent, preferably lactoferrin, the needy in this subject.

The present invention also relates to a method of increasing the sensitivity of a tumor of a subject to treatment of malignant tumors, including oral or parenteral administration of milk fat or similar milk fat, not necessarily at least the one additional drug, preferably, the lactoferrin in need of this subject separately, simultaneously or sequentially with the introduction of anti-tumor therapy. Preferably milk fat or similar milk fat is a described above. Preferably at least one additional drug is a described above. Preferably lactoferrin is a described above. Preferably the treatment is one of the above.

Equally the present invention also relates to a method for accelerating the recovery of the subject, the past treatment of malignant tumors, including the introduction of milk fat or similar milk fat, optionally at least one additional pharmaceutical agent, preferably lactoferrin, the needy in this subject separately, simultaneously or sequentially with the introduction of the treatment. In this embodiment of the invention the subject recovered from the effects of a malignant tumor or the treatment of malignant tumors more rapidly than the subject that is not treated according to the invention. State of a subject, preferably enables you to reduce the dose or time of treatment of malignant tumors.

These methods can be combined with treatments using Liu is passed one or more anticancer agents (including chemotherapeutic drugs or immunotherapy) or anticancer treatments above.

In one embodiment, the implementation of the milk fat or similar milk fat, optionally at least one additional drug, entered daily for at least about 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 weeks before the introduction of antineoplastic or anticancer treatment. In one embodiment, the implementation of the milk fat or similar milk fat, optionally at least one additional drug is introduced within at least approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or 21 days, or for at least about 1, 2, 3, 4, 5, 6, 7 or 8 weeks, or for at least about 1, 2, 3, 4, 5 or 6 months before the introduction of antineoplastic or anticancer treatment. In one embodiment, the implementation of the milk fat or similar milk fat, optionally at least one additional drug is introduced within at least approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or 21 days, or for at least about 1, 2, 3, 4, 5, 6, 7 or 8 weeks, or for at least about 1, 2, 3, 4, 5, or 6 months after the start of injection of antineoplastic or anticancer treatment.

13. The types of tumors

In one the m variant implementation of the tumor, cells of the tumor or a malignant tumor are leukemia, carcinoma of the colon, breast cancer, melanoma, skin cancer, or lung. The types of tumors that are relevant to the present invention are listed in the published international patent application WO 2006/054908, which is incorporated herein by reference.

In one embodiment, the implementation of the tumor cells of the tumor or a malignant tumor are leukemia, such as, but not limited to them, acute leukemia, acute lymphocytic leukemia, acute granulocytic leukemia, acute military leukemia, such as myeloblastic, promyelocytic, myelomonocytic, monocytic, erythroleucus and myelodysplastic syndrome, chronic leukemia, such as, but not limited to them, chronic miliitary leukemia, chronic granulocytic leukemia, chronic lymphocytic leukemia and hairy cell leukemia.

In one embodiment, the implementation of the tumor cells of the tumor or a malignant tumor are a lymphoma, such as, but not limited to it, Hodgkin's disease and sickness nehodzhkin. In one embodiment, the implementation of the tumor is the tumor cells originate from or malignant tumor includes hematopoietic tumor of myeloid series, such as, but not limited to it, sharp and XP the technical myelogenous leukemia, smoldering multiple myeloma, non-secretory myeloma and osteosclerotic myeloma. In one embodiment, the implementation of the tumor is the tumor cells originate from the lymphoid series or malignant tumor includes hematopoietic tumor of lymphoid series, including leukemia, acute and chronic lymphocytic leukemia, acute and chronic lymphoblastic leukemia, B-cell lymphoma, T-cell lymphoma, lymphoma Bernita. In one embodiment, the implementation of the tumor is the tumor cells originate from or malignant tumor includes hematopoietic tumor B-cell lymphocytic series. In one embodiment, the implementation of the tumor is the tumor cells originate from or malignant tumor includes hematopoietic tumor T-cell lymphocytic series.

Additional malignant tumors and related diseases that can be treated or prevented by methods and compositions of the present invention, include as non-limiting examples of the following: leukemia; lymphoma; multiple myeloma; macroglobulinemia waldenstrom; monoclonal gammopathy of undetermined significance; benign monoclonal gammopathy; heavy chain disease; bone sarcoma and connective tissue; brain tumor; breast cancer is elez; malignant tumor of the adrenal gland; thyroid cancer; malignant pancreatic tumor; malignant tumor of the pituitary gland; malignant neoplasm of eye malignant neoplasm of vagina; malignant tumor of the female external genital organs; malignant cervical tumors; malignant uterine tumors; malignant tumor of ovary; malignant tumor of the esophagus; malignant tumor of stomach; malignant tumor of colon; malignant tumor of the rectum; malignant liver tumor; malignant tumor of the gallbladder; cholangiocarcinoma; malignant lung tumors; testicular a malignant tumor; malignant tumor of the prostate; kidney a malignant tumor; malignant tumor of the oral cavity; basal cell carcinoma; malignant tumor salivary gland; malignant tumor of the throat; malignant tumor of the skin; malignant kidney tumor; Wilms tumor and a malignant tumor of the bladder. For a review of such disorders, see Fishman at al., 1985, Medicine, 2d Ed., J.B. Lippincott Co., Philadelphia and Murphy at al., 1997, Informed Decisions: The Complete Book of Cancer Diagnosis, Treatment, and Recovery, Viking Penguin, Penguin Books U.S.A., Inc, United States of America.

Additional malignant tumors or other abnormal proliferative diseases which may be treated or prevented according to the invention, include as non-limiting examples, the following: carcinoma, including carcinoma of the liver, spleen, heart, lung, small intestine, large intestine, rectum, kidneys, brain, bladder, breast, colon, kidney, liver, lung, ovary, pancreas, stomach, cervix, thyroid and skin; including squamous cell carcinoma; hematopoietic tumors of the lymphocytic series, including leukemia, acute lymphocytic leukemia, acute lymphoblastic leukemia, B-cell lymphoma, T-cell lymphoma, lymphoma Berkata; hematopoietic tumors of myeloid series, including acute and chronic myelogenous leukemias and promyelocytic leukemia; tumors of mesenchymal origin, including fibrosarcoma and rhabdomyosarcoma; other tumors, including melanoma, seminoma, teratocarcinoma, neuroblastoma and glioma; tumors of the Central and peripheral nervous system including astrocytoma, neuroblastoma, glioma and sandamu; tumors of mesenchymal origin, including fibrosarcoma, rhabdomyosarcoma, and osteosarcoma; and other tumors, including melanoma, pigmented the xeroderma, keratoakantoma, seminoma, thyroid follicular cancer cancer and teratocarcinoma.

In specific embodiments, implementation, malignant or deproliferation changes (such as metapl the AI and dysplasia), or hyperproliferative disorders treat or prevent the ovaries, bladder, breast, colon, liver, lung, skin, pancreas, or the uterus. In other specific embodiments, implementation, cure or prevent sarcoma, melanoma or leukemia.

14. Treatment or prevention of malignant tumors of the skin

In an additional embodiment, the invention presents a method of treating or preventing malignant tumors of the skin, including the stage of the introduction of milk fat or similar milk fat, optionally at least one additional pharmaceutical agent, preferably lactoferrin, in or on the skin and/or in the region of the tumor.

In the preferred embodiment, due to the impact of solar radiation skin prone to malignant tumors of the skin.

In a preferred embodiment, a malignant tumor is basal cell carcinoma, squamous cell carcinoma or melanoma.

Preferably, the milk fat or similar milk fat injected locally, both individually and in combination with standard treatment regimen for malignant tumors. Introduction in the area of the tumor includes the introduction in the vicinity of or near the boundaries of the tumor or directly in the area of the tumor. It is assumed that milk fat inhibits carcinogenesis, stimulates local protivoop alevy immunity in tissues, inhibits tumor angiogenesis and/or is a means of directly destroying tumor cells can inhibit tumor growth). In summary, milk fat, optionally at least one additional pharmaceutical agent in a suitable medium in concentrations of 0.1%, 1%, 5% or 10% applied twice daily to the skin in the risk zone or area of skin affected by a malignant tumor. The progression of tumor size is controlled by means of images obtained with CT and tumor markers, where available.

About doses and treatment regimens can learn by conducting preclinical studies on a suitable model of malignant skin tumors in animals. The area of skin in mice are shaved and treated topically with the use of a carcinogen (e.g., 7,12-dimethyl-Benz[a]anthracene (DMBA), then perhaps the subsequent exposure to UV-B (Bestak, et al., 1996). Lactoferrin can be applied within two days after application of the carcinogen or as soon as formed, the lesion is malignant, preferably in the presence of substances that promote penetration into the skin (such as 70% Laureth sulfate and 30% phenylpiperazine), which are able to increase skin permeability. Lactoferrin is applied twice a day or otherwise, if necessary, on the skin or lesion malignant SDA is the role and control of tumor growth over a period of weeks to months.

If necessary, these methods can be combined with methods of treatment using any one or more anticancer agents (including chemotherapeutic drugs or immunotherapy) or anticancer treatments described above.

15. Anemia

As shown in the examples below, oral administration of milk fat is effective to reduce hematologic suppression in the subject and treating or preventing anemia in a subject.

Anemia refers to a deficiency of red blood cells (RBCs) and/or hemoglobin. The result of this reduced ability of blood to transfer oxygen to the tissues, which leads to hypoxia. Thus, different stages of anemia can lead to a wide range of clinical consequences.

Using a morphological approach, anemia can be classified by the size of erythrocytes. The size is reflected in the average volume of erythrocytes (MCV), which is measured, usually in femtoliter (fl). If the cells are smaller than normal (up to 80 fl), the anemia is called microcytic; if the cells are normal in size (80-100 fl), normocytes; and if the cells are larger than normal (>100 fl), the anemia is classified as macrocytic.

Microcytic anemia is primarily a result of violations in the synthetic is e or insufficient hemoglobin hemoglobin synthesis and can be caused by several reasons, including defects in heme synthesis, iron deficiency, anemia of chronic disorders, defects in the synthesis of globin, alpha - and betathalassemia, HbE syndrome, HbC syndrome and other diseases associated with the presence of unstable forms of hemoglobin defects sideroblasts, including hereditary sideroblastic anemia, acquired sideroblastic anemia, including from the toxic action of lead and reversible sideroblastic anemia. Zhelezodificitnaya anemia is the most widespread type of anemia, and it is due to many reasons. When viewed under a microscope is often revealed hypochromic (paler than usual) and microcytic (smaller than normal RBCs. Zhelezodificitnaya anemia is a condition, usually caused by inadequate intake of iron from food or insufficient absorption of iron. Zhelezodificitnaya anemia can also be caused by bleeding from damaged areas of the gastrointestinal tract. As microcytic anemia usually classified hemoglobinopathy, such as sickle cell disease and thalassemia.

Normalfree anemia is a condition in which the total Hb level decreased, but the size of erythrocytes (MCV) remains normal. Causes include acute blood loss, anemia of chronic disease, plastices the Yu anemia (violation of bone marrow hematopoiesis) and hemolytic anemia. Hemolytic anemia caused by a separate set of symptoms with a number of possible reasons (also characterized by jaundice and elevated LDH level). The disease may be an autoimmune, immune, hereditary or be a result of mechanical interference (e.g., surgery on the heart). Although often normalfree, it can occur as a result of cell disruption microcytic anemia or due to premature release of immature erythrocytes from the bone marrow macrocytic anemia.

Macrocytic anemia can be further divided into "megaloblastic anemia" or "semiglobata macrocytic anemia". A cause of megaloblastic anemia is primarily a violation of DNA synthesis with saved RNA synthesis, which leads to limited cell division of progenitor cells. Megaloblastic anemia is often presented with hypersegmented neutrophils (6-10 shares). Megaloblastic anemia is the most common case macrocytic anemia. Megaloblastic anemia occurs mainly because of the shortage of both vitamin B12 and folic acid (or both), which in turn usually occurs or due to inadequate intake or poor absorption. Folate deficiency in normal does not lead to neurological is the how do I get it, unlike B12 deficiency. Pernicious anemia is an autoimmune condition that is directed against parietal cells of the stomach. Parietal cells produce internal factor needed for absorption of vitamin B12 from food. Thus, the destruction of parietal cells leads to a lack of intrinsic factor, which leads to low absorption of vitamin B12. Megaloblastic anemia can also be the result of exposure to methotrexate, zidovudine, and other drugs that inhibit DNA replication. Amaglobeli macrocytic anemia have other reasons (i.e. undamaged DNA synthesis), which manifest themselves, for example, in alcoholism.

There are various specific anemia, including anemia of prematurity, which occurs in premature infants within 2 to 6 weeks after birth and is the result of decreased erythropoietin response to decreasing levels of hematocrit; anemia Fanconi's syndrome, hereditary disorders or defects, characterizing aplastic anemia and various other disorders; hereditary spherocytosis, hereditary disorder that leads to defects in the membrane of erythrocytes (RBC), which leads to the deposition and destruction of erythrocytes in the spleen. This leads to a decrease in the number of circulating RBCs is, thus, to anemia; sickle cell anemia, a hereditary disorder that occurs due to the presence of a mutant gene for hemoglobin S; autoimmune hemolytic anemia with heat agglutinins, anemia caused by autoimmune attack against red blood cells, primarily by IgG; and hemolytic anemia associated with holomovie agglutinins mainly mediated by IgM.

Thus, the present invention relates to a method of treating or preventing anemia in a subject comprising administration to the subject of milk fat or similar milk fat optionally at least one additional drug. The present invention also relates to methods of reducing or inhibiting hematologic suppression of the subject.

The reduction or inhibition of hematologic suppression has application not only in subjects undergoing treatment, or those who have passed anti-cancer treatment, but also in other disorders, including as non-limiting examples of anemia described above. Therefore, milk fat or similar milk fat, preferably in combination with at least one additional drug, more preferably lactoferrin or a functional variant or functional fragment and especially l is ceferina, containing metal ion, have application outside the field of treatment and prevention of malignant tumors.

16. Cachexia

As shown in the examples below, oral administration of milk fat is effective to improve the condition of the subject and for treating or preventing cachexia in a subject.

Cachexia are one or more signs, it is weight loss, muscle atrophy, fatigue, weakness and lack of appetite (significant loss of appetite in someone who is not committed to an intensive weight loss. This condition is usually associated with underlying disease, such as malignant tumor, certain infectious diseases (e.g. tuberculosis, AIDS) and some autoimmune disorders. Cachexia physically weakens subjects to a state of immobility caused by loss of appetite, asthenia, and anemia and is usually a response to standard treatment is weak.

Subjects with malignant tumor cachexia caused by both reduced physical activity, joined infections, and toxic effect of chemotherapy and radiotherapy on the digestive tract, usually are symptomatic treatment to maintain the quality of food and quality of life. Such regulation includes the use of rinse mouth with stomatitis is, frequent consumption of foods in small portions, antiemetics, antibiotics, transfusion of blood and oral and parenteral nutritional supplements. Dietary supplements can be effective to provide additional calories, proteins, fats, vitamins and minerals. In specific cases, such as malabsorption syndrome on the background of carcinoma of the pancreas, used exogenous extract of the pancreas to improve absorption of fat and protein (Perez MM, et al. Evaluation of weight loss, food intake, fat metabolism, malabsorption and treatment of pancreatic insufficiency in cancer of the pancreas. Cancer 1983; 52:346-52).

Various pharmacological tools were introduced when trying to pay, delay or halt progression of cachexia in subjects with malignant tumor. They include corticosteroids, such as prednisolone, methylprednisolone and dexamethasone, progestogenic tools, such as megestrol acetate, medroxyprogesterone acetate, cannabinoids such as dronabinol, serotonin antagonists such as cyproheptadine at tools that affect intestinal motility, such as metoclopramide and cisapride, steroidal anabolic agent, such as nandrolone decanoate and fluoxymesterone, inhibitors phosphoenolpyruvate-carboxykinase, such as the guide is azina sulfate, analogues of methylxanthines, such as pentoxifylline and lisofylline, thalidomide, cytokines and anticytokines, such as antibody Anti-IL-6, IL-12, branched chain amino acids, eicosapentaenoic acid, inhibitors of prostaglandin synthesis, such as indomethacin and ibuprofen, hormones such as melatonin and agonists β2-adrenergic receptors, such as clenbuterol.

Thus, the present invention relates to a method of treating or preventing cachexia in a subject, comprising administration to the subject of milk fat or similar milk fat, optionally, at least one additional drug. The present invention also relates to a method of improving the condition of the subject with cachexia.

Treatment or prevention of cachexia has application not only in subjects undergoing treatment, or those who have passed anti-cancer treatment, but also in other disorders associated with weight loss or fatigue, including as non-limiting examples of disorders described above. Therefore, milk fat or similar milk fat, preferably in combination with at least one additional drug, more preferably lactoferrin or a functional variant or functional fragment and particularly those containing metal ion, lactoferrin who have applications outside the treatment and prevention of malignant tumors.

17. Mucositis

As shown in the examples below, oral administration of milk fat is effective for improvement in the damage of the digestive tract, such as ulcers and in the treatment or prevention of mucositis.

Mucositis is a condition that is characterized by damage to the epithelium of the oral cavity, and pharynx and gastrointestinal tract (GI), resulting in inflammation and ulceration of these mucous membranes. In the oral cavity and esophageal mucositis is characterized by the formation of painful ulcers. Further along the digestive tract mucositis causes diarrhea, which is often heavy and exhausting. Mucositis is usually the result of radiation and/or chemotherapy, which occur in varying degrees, approximately 40% of subjects who received chemotherapy against malignant tumors. Due to the relatively high refresh rates compared to cells in other organs, epithelial cells are more sensitive to the cytotoxic effects of radiotherapy and chemotherapy. In most cases, the epithelial cells of the mucous membranes have a faster update than to be treating a malignant tumor, and are vulnerable to damage cytotoxic means and irradiation. Diagnosis and control of torment is Zita successfully through survey of the subject (such as a questionnaire on pain), inspection of the oral cavity and endoscopy. Also recommended some tests, to be determined for the diagnosis, including a breath test for sucrose, citrulline and test transglutaminase.

Treatment of mucositis is mainly supportive. Currently the mainstay of treatment is oral hygiene achieved, usually through frequent rinsing of the mouth. For lubrication of the oral cavity can be used water-soluble gels. To relieve pain and increased sensitivity, which is associated with mucositis, there are various local palliative means. Rinsing the mouth with saline can help alleviate the pain and remove particles of food product in order to avoid infection. Medicines include chlorhexidine gluconate, lidocaine (Xylocaine), dyclonine HCl (Declan) and benzocaine in the composition Arabas (Orabase). Additionally, diphenhydramine HCl (Benadryl), which has a local anesthetic effect, can be mixed in equal volumes, or Kaopectate (Kaopectate) or magnesium hydroxide with formation of a suspension. The benzydamine hydrochloride (HCl) is a nonsteroidal drug for rinsing with an anti-inflammatory, analgesic and anesthetic properties. A number of researchers have described the use of sucralfate, widely introduced the CSOs in the treatment of gastric ulcer, in the form of mouthwashes in the treatment of mucositis caused by radiation therapy and chemotherapy.

It has been described that a bactericidal agent, such as chlorhexidine gluconate, polymyxin E, tobramycin and amphotericin, have some clinical significance. It has been described that anti-inflammatory agents such as betamethasone and indomethacin, potentially important in preventing or reducing the severity of mucositis caused by radiation. Described the use of palifermin (KEPIVANCE), human keratinocyte growth factor (KGF), to enhance the proliferation, differentiation and migration of epithelial cells. Other methods of treatment based on the use of cytokines and other modulators of inflammation (such as IL-1, IL-11, TGF-beta), supplements with amino acids (e.g. glutamine), vitamins, colony stimulating factors, such as colony-stimulating factor for granulocytes and macrophages (GM-CSF), cold treatment and laser treatment.

Thus, the present invention relates to a method of treating or preventing mucositis in a subject comprising administration to the subject of milk fat or similar milk fat, optionally, at least one additional drug. The present invention also relates to a method of improving the condition of the subject when mucositis.

Leche is and / or prevention of mucositis and damage reduction or improvement in injuries of the gastrointestinal tract has application not only in subjects, treatment, or those who have passed anti-cancer treatment, but also in other disorders associated with damage to the epithelium of the digestive tract, including as non-limiting examples of disorders described above. Therefore, milk fat or similar milk fat, preferably in combination with at least one additional drug, more preferably lactoferrin or a functional variant or functional fragment and particularly those containing metal ion-lactoferrin has application outside the treatment and prevention of malignant tumors.

18. Leukocytopenia

As shown in the examples below, oral administration of milk fat is effective to reduce hematologic suppression in subject and in treatment and prevention of radiation from the subject.

Leukocytopenia (also known as leukopenia and including state, sometimes referred to as lymphopenia or lymphocytopenia) refers to the reduction in the number of leukocytes (WBCs) in the subject. This leads to a reduced ability of the body to resist infections, which leads to greater vulnerability of the body. Different degrees of radiation can thus be equated to a wide range of clinical consequences.

On average, an adult has the WBC quantity of 500-10000 cells/cubic millimeter (varies depending on gender and subject). Leukopenia determine, as a rule, when the number of WBC less than 4000 cells/cubic millimeter (again varies from the floor and subject). The two main forms of leukocytopenia are neutropenia and granulocytopenia.

Neutropenia is a major leukocytopenia and is characterized by a reduced number of neutrophils. In addition to malignant tumors, many syndromes associated with neutropenia, most of which is hereditary in nature. Kostmann neutropenia syndromes Shwachman, agammaglobulinemia, dysgammaglobulinemia, myelokathexis syndrome krasivo-hair hypoplasia and congenital diskeratoz are just a few examples of disorders associated with neutropenia. Neutropenia is also associated with deficiencies of nutrients such as vitamin B12, folic acid or a lack of copper.

The muscle problems are sometimes used interchangeably with neutropenia, but more accurately granulocytopenia is a drop in the number of eosinophils and basophils, and neutrophils.

Leukocytopenia can also occur when subjects are exposed to chemotherapy or radiation therapy. It is a common side effect and subjects usually given time to recover and education the larger the number is TBA WBCs before continuing therapy. Basically, the radiation is usually treated with steroids or vitamins to stimulate the bone marrow to produce more WBCs.

Thus, the present invention relates to a method of treating or preventing radiation from the subject, comprising administration to the subject of milk fat or similar milk fat optionally at least one additional drug. The present invention also relates to a method of reducing or inhibiting hematologic suppression of the subject.

The reduction or inhibition of hematologic suppression has application not only in subjects undergoing treatment, or those who went through cancer treatment, but also in other disorders, including as non-limiting examples of leukocytopenia above. Therefore, milk fat or similar milk fat, preferably in combination with at least one additional drug, more preferably lactoferrin or a functional variant or functional fragment and particularly those containing metal ion-lactoferrin has applications outside the treatment and prevention of malignant tumors.

Various aspects of the invention will now be illustrated in the way, but unlimited by them, by reference to the following examples, the am.

EXAMPLES

Mouse and reagents

Worked with female C57BL/6 and mice of Balb/c at the age of six to nine weeks (University of Auckland, New Zealand), and in each group on nutrition included 5 or 6 mice, unless otherwise noted. Mice were kept in a room with air-conditioning with adjustable humidity, temperature and 12 h cycle of alternation of light and darkness. Murine T-cell lymphoma of the thymus EL-4 from C57BL/6 and cancer cell line 4T1 mammary glands of BALB/c mice (H-2b) were purchased from American Type Culture Collection (Rockville, MD, USA). Cells, EL-4 were cultured at 37°C in DMEM (Gibco BRL, Grand Island, NY, USA), whereas the 4T1 cells were maintained as monolayer cultures at 37°C in RPMI medium 1640 (Gibco BRL, Grand Island, NY, USA). Wednesday was further added 10% fetal calf serum, 50 U/ml penicillin/streptomycin, 2 mm L-glutamine and 1 mm pyruvate. Paclitaxel was purchased from Bristol-Meyers Squibb, WA, USA. All experiments were performed under a Protocol approved by the Animal Ethics Committee, University of Auckland.

Getting milk fat

Milk fat enriched with conjugated linoleic and vaccinology acids, obtained through supplementary feeding of free fatty acids of cows on pasture, according to the method Harfoot et al. 58. Normal anhydrous milk fat was obtained from Fonterra Co-operative Group Limited, NZ. Composition of milk fat and enriched milk fat used in therapeutic diets, generalization is s in tables 6a and 6b. The data in tables 6a and 6b were obtained using FAMES, advanced FAMES, CLA and analyses of milk fat, well-known in this field.

Table 6a
The content of milk fat in the diets
CLA componentNormal (% wt./mass.)Enriched (% wt./mass.)The magnification of
CLA (C-9,t-11)1,175,043,3
CLA t-10,c-12)--
CLA (minor c18:2 isomers)0,551,301,4
Total CLA (all forms)1,726,342,7
The ratio of c-9, t-11 CLA to the General CLAs68,0%79,5%

1,8
Table 6b
The content of milk fat in the diets
component fatty acid Normal (% wt./mass.)Enriched (% wt./mass.)The percentage reduction
c4:0 (butyric acid)3,23,2
c6:0 (Caproic acid)2,31,726%
c8:0 (Caprylic acid)1,3123%
c10:0 (capric acid)2,82,221%
c10:1 (2-decanoate)0,30,233%
c12:0 (lauric acid)3,22,522%
c12:1 (11-dolezelova acid)0,20,150%
c13:0 times. (tridecanoate acid-fu.)0,10100%
c13:0 (tridecanoate acid)0,10,1
c14:0 times. (myristic acid again.)0,20,150%
c14:0 (myristic acid)10,99,117%
c14:1 (myristoleate acid)0,90,811%
c15:0 ISO-fu.0,40,325%
c15:0 antiso times.0,60,6
c15:0 (pentadecanoic acid)1,31,28%
c 16:0 times.0,20,2
c16:0 (palmitic acid)30,619,736%
c16:1 (palmitoleic acid)3
c17:0 ISO-fu.0,60,6
c17:0 antiso times.0,40,5
c17:0 (margaric acid)0,80,538%
c17:10,30,3
c18:0 (stearic acid)10,54,656%
c18:1 n-9 (oleic acid)16,611,928%
c18:1 n-7 (Aksenova acid)the 4.722,9
c18:2 n-6 (linoleic acid)1,42,1
c18:3 n-30,80,450%
c18:2(CLA)1,25,3
c18:4 + isomers CLA00
c20:0 (orhideea acid)0,20,150%
c20:1 n-110,10,1
c20:1 n-9 (Aksenova acid)00,1
c20:2 n-600
c20:3 n-3 (eicosatrienoic acid)0,10100%
c20:4 n-6 (arachidonic acid)0,10100%
c20:3 n-300,1
c20:4 n-3 (eicosatetraenoic acid)0,10,2
c20:5 n-3 (eicosapentaenoic acid, EPA)0,10,2
c22:0 (Baganova acid)0,10,1
c22:1 n-13, n-11 (docosanoate acid)00,1
c22:2 n-900
C22:4 n-600
C22:5 n-3 (docosapentaenoic acid)0,10100%
C24:0 (Ognianova acid)00
c22:6 n-3 (DHA)00
c24:100
Fatty acids were identified through the om analysis of FAME. The fold increase relates to increased levels of lipid-enriched milk fat compared with milk fat. c18:1 n-7 provides an assessment of the content of TRANS c18:1 fatty acids.

Receiving lactoferrin

Bovine lactoferrin was obtained from skim milk (Fonterra Co-Operative Group Limited, New Zealand)using the method Norris et al. (Norris, GE et al., 1989). Ion-exchange resin SP Big Beads were loaded with skim milk and washed with water. The elution was performed with a column 0-0,5 M NaCl and the eluate is not used. Then the elution from the column was carried out with 0.5 to 1.0 M NaCl and collected eluate. The collected eluate was subjected to UF/DF using a membrane KD to reduce the amount of salts and components with low molecular weight. The filtration was continued up until retentate did not constitute from 90 to 93% of bovine lactoferrin. The extract obtained lactoferrin had a natural saturation levels of iron, about 15%, and is designated as bLf in the following examples. Extract of bovine lactoferrin saturated with iron (100% saturated), were obtained from natural bLf method of Law et al., (Law and Reiter, 1977), denoted as Lf+ in the following examples.

Diet

Experimental food rations received by Crop & Food Research, Palmerston North, New Zealand, using as a base powder composition AIN93G. In the AIN93G diet that does not contain lactoferrin, as the source of protein used casein is soybean oil as a source of lipids. In the experimental dietary casein was replaced natural bLf or Lf+, obtained as described above, the total protein content in the diet, therefore, was unchanged. The diet consisted of 28 g bLf, which is rich in iron, or 28 g of extract natural bLf 2.4 kg of food. Soybean oil was replaced in the experimental food rations or enriched milk fat, or normal anhydrous milk fat, obtained as described above, thus, the total fat content in the diet was unchanged. Fresh food was provided twice a week, and the mice had free access to food and water throughout the study. The phospholipid fraction of the Phospholipid concentrate Phospholac 600™ (PC600™) was purchased from Fonterra Co-operative Group Limited, New Zealand.

Experimental models of tumor and treatment

Tumors were inoculated by s.c. injection of cells, EL-4 at 2×105or 4T1 2×104in the left side of the mice, and tumor growth was evaluated by finding two perpendicular diameters. When the tumors had reached a size of more than 1.0 cm in diameter, animals were slaughtered in accordance with Animal Ethics Approval (University of Auckland). In all experiments consisted of 5 or 6 mice per treatment group, if not stated otherwise. Paclitaxel (30 mg, dissolved in 5 ml of Cremophor® EL and digidratirovannogo alcohol) was diluted with 0.9% NaCl and injected I.P. Pavlova. from R the account 30 mg/kg In the case of a metastatic model of breast cancer line 4T1 at the end of the experiments after killing the mice were removed and weighed organs, including lungs, liver and spleen. Counted the number of metastases on the lung surface. Livers were fixed in 4% paraformaldehyde and did cross sections with a thickness of 10 μm to 5 different levels to cover the entire liver. The sections were stained with hematoxylin and eosin. Counted the number of metastatic nodes, containing more than 6 malignant cells, and take the average value to represent the number of metastases. Blood was collected by cardiac puncture at the time of autopsy and counted on hemocytometer the number of blood cells.

Quantification of the production of antitumor cytotoxic T lymphocytes (CTLs)

Splenocytes were collected on certain days after injection of tumor cells. Incubated at 37°C with target cells, EL-4 gradient in the ratio E:T in 96-well round bottom plate. After 4 h incubation were collected and 50 μl of the supernatant fluid was measured by lysis of the cells using the Cyto Tox 96 Assay Kit (Promega, Madison, WI, USA). Included control measure background of non-specific lysis of target cells and effector cells. After subtracting the background was estimated percentage of cell lysis, using the formula: 100 × (experimental spontaneous shall effector - spontaneous target/maximum target - spontaneous target).

Quantification of apoptosis

Tumors were excised and immediately frozen in dry ice and kept at -70°C for further detection of apoptotic cells in tumors in situ. Frozen series of slice thickness of 6 μm were fixed in a solution of paraformaldehyde (4% in PBS, pH 7,4) and violated the permeability of cell membranes with a solution containing 0.1% Triton X-100 and 0.1% sodium citrate. Slices were incubated with 20 μl of TUNEL reagent (commercially available kit for determination of apoptosis in situ from Boehringer Mannheim, Germany) for 60 min at 37°C and examined by fluorescence microscopy. Adjacent contrasting sections were stained with hematoxylin for counting the total number of cells or the number of apoptotic cells in ten areas selected randomly (magnification x40). Apoptotic index (AI) was considered as the number of apoptotic cells × 100/total number of nucleated cells. To identify apoptotic cells in vitro, the number of apoptotic and necrotic tumor cells was measured by staining with fluorescence method with annexin V, TUNEL and Trifanova blue. To measure apoptosis in the digestive tract, sections of small intestine were stained by the TUNEL method using the DeadEndTM Fluorometric TUNEL System (Promega, Madison, WI, USA). Calculated cycloheptadiene cells 10 crypts, selected randomly and data were presented as the number of apoptotic Taurus on the crypt.

Assessment of vascularization

For determination of tumor vascularization, frozen tumor sections with a thickness of 10 μm were fixed in acetone, washed with PBS, blocked with 2% BSA for 2 h and was tuberculi overnight with the antibody MEC13,3 anti-CD31 or mAb against CD105. The sections then were incubated for 30 min with secondary antibodies, using a set of VECTASTAIN Universal Quick kit (Vector Laboratories, Burlingame, CA, USA); and treated with Sigma FAST DAB (3,3'-diaminobenzidin tetrahydrochloride) and pills that increase CoCl2(Sigma) and contrast were stained with hematoxylin. Stained blood vessels were counted under x40 magnification in five or six areas (0,155 mm2), blindly selected randomly, and calculated the average of the three maximum values. To monitor blood flow in the blood vessels in the tail vein injected DiO7 at a concentration of 1.0 mg/kg (Molecular Probes, Eugene, OR) for one minute to collect tissues, as described previously (Ding et al., 2001). To assess vascularization also used the method of concentric circles.

Assessment cachexia

Mice were weighed at the beginning and at the end of the experiment. After mice were killed, and tumors were excised and weighed. To determine the amount of adipose tissue and muscle weakness was dissected epididymal W the global fabric and left gastrocnemius muscle. Slaughter weight was calculated as the difference in weight between the total weight and weight of tumors.

Quantitative evaluation of damage of the digestive tract

Skinny intestine of mice were fixed in 4% paraformaldehyde, were immersed in paraffin, did the slice thickness of 4 μm, and the sections were stained with hematoxylin-eosin. Each sample was chosen randomly 20 well preserved villi and measured their length by microscopy, using an objective micrometer.

Recorded the activity of γ-glutamyltranspeptidase (γ-GGT) in the jejunum, as a separate indicator of damage to the intestine, as described previously (Ziotnik et al., 2005). Part of the jejunum (approximately 5 cm) was separated and washed with 10 ml PBS and cut into two pieces 2 cm2that was placed in 1.0 ml of 1.0% Triton X-100, 0.15 M NaCl, 100 mm Tris, pH 8.0 (Tris buffer). Added twenty microlitres solution containing 0.3 ml of 100 mm glycyl-glycine, pH 8.0, and 0.08 ml of Tris buffer, 0.5 ml of 5 mm γ-glutamyl-p-nitroanilide, to a final volume of 0.9 ml Portion of the digestive tract were incubated for 10 min in a water bath, with stirring, at 37°C and recorded the activity of γ-GGT at 405 nm. The results were expressed as units of activity of γ-GGT on cm jejunum, where one unit is defined as the activity of γ-GGT, which releases 1.0 mol of p-nitroaniline at 1 o'clock

Statistical analysis

The results presented and as mean ±SEM or confidence interval ±95% and statistical significance was assessed, using the student's t test (examples 1 to 17), or analysis of variance or analysis of covariance, followed by the method of multiple comparison Tukey's, or 1-sided test Dannette (examples 18 to 21). The value of P<0,05 indicates statistical significance, whereas P<0,001 shows that the results are highly significant.

Example 1

Bovine lactoferrin with a purity of more than 90% was purchased from Fonterra Co-operative Group. To get APO-Lf, about 80 mg/ml Lf in milliQ water (pH~5,7), brought to pH 2,08 careful addition of 6 M HCl. The solution was stirred at RT for 1 h, and then were dialyzed overnight at 4°C against 10 volumes of 0.1 M citric acid, using SpectraPor dialysis tube with a nominal cutoff molecular weight of 3.5 kDa (Spectrum Companies, Ranco Dominguez, CA, USA). The dialysis solution was changed twice during the 24 h period and the solution Lf liofilizirovanny to white semi-crystalline powder. To obtain lactoferrin saturated with Fe 50%, 8% solution of lactoferrin in 0.1 M sodium bicarbonate was brought to a pH of 8.2 careful addition of 6 M NaOH. An appropriate volume of 50 mm nitrilotriacetate iron (Fe-NTA) (Bates et al., 1967; Brock & Arzabe, 1976) were added to obtain ~50% saturation of lactoferrin (taking into account the purity of the Lf and its natural saturation of Fe ~12%). After stirring for 1 h at RT, the solution (pH 8,01) were dialyzed overnight at 4°C against 10 volumes of water milli-Q, IP is by using SpectraPor dialysis tube, as explained above. The dialysis solution was changed twice during the 24 h period and the solution Lf liofilizirovanny to salmon-red semi-crystalline powder. Lactoferrin with ~100% saturation Fe received basically the same as for substance, saturated Fe 50%, except for the adjustment of an appropriate number of Fe-NTA and the subsequent addition of Fe-NTA, peredvigenija pH to 8.0 by careful addition of 6 M NaOH. The final product was a dark salmon-red semi-crystalline powder. The saturation levels of Fe end-products was checked by spectrophotometric titration (Bates et al., 1967; Brock & Arzabe, 1976). APO-lactoferrin was saturated Fe approximately 5%.

Example 2

The example shows that milk fat inhibits the growth of tumors EL-4, whereas milk fat enriched with conjugated linoleic and vaccinology acids is ineffective.

A group of six C57BL/6 contained on the control diet AIN-93 or the same diet with the replacement of 120 g or milk fat, or enriched milk fat by 2.4 kg of food, providing ~71% of the fat component of the diet. Two weeks after the beginning of such a diet mice were inoculated subcutaneously with 2×105tumor cells, EL-4. Tumor size was measured by finding two perpendicular diameters (in cm) was controlled to 91 days or until it reaches distend the pour of size 1 cm in diameter. Each dot represents the average tumor size with a confidence interval of 95%, as specified 6 mice, and for the indicated number of mice.

Enriched milk fat slowing the growth of tumors in 25% to 49 days, compared with the control diet, but the effect was insignificant (figure 1). In contrast, marked milk fat completely prevented the development of tumors in 2 of 6 mice. Tumor growth in the other 4 mice that were kept on a diet with milk fat was similar to the growth of tumors in mice that were fed enriched milk fat.

Example 3

The example shows that milk fat acts synergistically with immunotherapy in the destruction of tumors EL-4.

Tumors were grafted in the group of five mice, which were fed as enriched milk fat, milk fat, and maintained on the control diet, as described above in example 2. Controlled tumor size, measured by finding two perpendicular diameters (in cm). When the tumors ~0.4 cm in diameter, the tumor was injected complexes DNA-liposomes containing 60 µg expressing plasmids B7-1. The moment of introduction of the plasmids shown by the arrow. Tumor size was measured by finding two perpendicular diameters (in cm), controlled before 91 days or until the tumor size 1 cm in diameter. Tumor this is a specific size partially retain sensitivity to immunogenic treatment of B7-1, as evidenced by the fact that the control tumors in four mice slowly decreased over the course of one week, but then resumed growth. However, the tumor in one mouse was decreased and completely disappeared within a 4 week period after the injection of plasmids B7-1 (figure 2).

Tumors in mice fed milk fat decreased more rapidly than in control mice, because they needed 2 weeks to extinction after the injection of plasmids B7-1. In contrast, tumors in mice fed enriched milk fat were resistant to immunogenic treatment of B7-1 and was growing at a rate close to the growth of tumors in mice that were kept on a control diet. Therefore, milk fat increases the effect of immunotherapy against malignant tumors, whereas enriched milk fat is less efficient.

Example 4

The example shows that milk fat acts synergistically with lactoferrin saturated with iron to completely inhibit the formation of tumors.

In previous studies, the applicants have shown that Lf saturated with iron (Lf+), able to inhibit tumor growth (WO/2006/054908). In these experiments investigated the effect of joint introduction of milk fat and Lf+. We tried to determine whether milk fat to act synergistically with Lf+ in the fight against lymphoma.

Groups is from six mice were kept on a control diet AIN-93G, diets containing 28 g Lf+, or 120 g milk fat (AMF) 2.4 kg of food, or a diet containing a combination and Lf, and milk fat. Day 0 refers to the day of beginning of the premises mice on a proper diet. After two weeks of diet in Boca mouse was injected with 2×105cells, EL-4. The tumor size is found by measuring two perpendicular diameters (in cm), controlled before 56 days. Each dot represents the average tumor size with a confidence interval of 95%, as specified 6 mice, and for the indicated number of mice.

The appearance of tumors was delayed by one week in 5 out of 6 mice fed Lf+, and all 6 mice, which were kept on the diet from milk fat (figa). One of the 6 mice, which were kept on a diet with Lf+, there is a complete tumor rejection. In contrast, a complete rejection of grafted tumors occurred in all six mice that were kept on a diet containing a combination of the Lf+ and milk fat, which indicates that milk fat acts synergistically with Lf+ in the destruction of lymphoma.

Example 5

The example shows that milk fat increases the ability Lf+ to increase the antitumor cytotoxicity of leukocytes and tumor apoptosis.

Splenocytes were isolated on the 56th day of the mice described in example 4, and analyzed their cytolytic activity against target cells, EL-4. The anti-Christ. puhaleva the cytolytic activity of splenocytes, obtained from mice that were fed or Lf+, or milk fat was significantly increased by 66% (P<0.001) and 61% (P<0,01), respectively, compared to mice, which were kept on a control diet (pigv). Antitumor cytolytic activity was also increased by 86% (P<0,001) compared to control mice fed a combination of Lf+ and milk fat.

Sections were obtained from tumors of mice described in example 4, at 56 days and were stained by the indirect way end deoxynucleotidyltransferase end labeling breaks desacetyldiltiazem-digoxigenin (TUNEL) and also by way of fluorescence with annexin V (annexin-V-fluos). The number of apoptotic cells was determined by TUNEL method or fluorescence with annexin V (annexin-V-fluos), were analyzed by 10 randomly selected areas of stained tumor sections, visualized through increased x40. Apoptotic index (A/I) is the number of cells in apoptosis (positive in the TUNEL or fluorescence with annexin)×(100/total number of cells). Milk fat and Lf+ stimulated tumor apoptosis by 73% and 68% respectively compared with the control diet (figs). In accordance with its high antitumor cytolytic activity of the combination of milk fat and Lf+ increased tumor apoptosis by 84%.

Example 6

what this example shows that milk fat inhibits tumor angiogenesis.

Effects Lf+ and milk fat on tumor blood flow and blood vessel formation was analyzed by staining of tumor sections obtained as described in example 5, with mAbs against CD31 and anti CD 105 and through the perfusion DiO7 respectively. From mice subjected to processing as described in example 4, at 56 day received tumor slices and stained or with mAb MEC13,3 against CD31, or mAb against CD 105 for visualization of blood vessels, or alternatively, one minute to collect tissues, into the tail vein injected DiO7 in order to visualize the blood flow. Colored blood vessels believed six mice in six blindly selected random fields.

As shown in fig.3D, the number of CD31 + vessels in tumors of mice fed Lf+, milk fat or a combination of milk fat and Lf+was significantly reduced by 84% (P<0,001), 72% (P<0.001) and 84% (P<0,01), respectively, and the blood flow was reduced by 84% (P<0,001), 68% (P<0.001) and 84% (P<0,001), respectively, compared with mice, which were kept on a control diet. Similar results were obtained when staining of CD105+ vessels and perfusion DiO7 as additional markers of formation of tumor vessels. These results show that milk fat inhibits angiogenesis.

Example 7

The example shows that the powder alpha lip is Dov (Phospholac 600™), sphingomyelin and 9, 11 CLA isomers inhibit tumor growth, but does not act synergistically with Lf+ for the effective destruction of tumors.

A group of five C57BL/6 contained on the control diet AIN-93 (figa) or the same diet with the replacement of 28 g Lf+ (pigv), 120 g phospholac 600™ (figs) or 1.2 g sphingomyelin (fig.4D) (providing ~71 and 0.71% of the fat component of the diet, respectively) 2.4 kg of food. Two weeks after the appointment of the diet, mice were inoculated subcutaneously with tumor cells, EL-4. Tumor size was again measured by finding two perpendicular diameters (in inches).

Lf+ delayed the tumor growth of two tumor growth was delayed by two weeks and one at least one week. Phospholac 600™ completely inhibited opukholeobrazovanie one mouse and delayed the formation of tumors in other mice within 24 hours, while had no significant effect on tumor growth in the other three mice compared with mice that were kept on a control diet. Sphingomyelin was as effective as and Lf+because delayed tumor growth by ~1 week from two mice and ~2 weeks the other two mice, while had no significant effect on tumor growth in other mice. An additional group of five mice fed Lf combinations+ (28 g per 2.4 kg of diet) and or Phospholac 600™ (120 g 2.4 kg food) (fige), and the and sphingomyelin (1.2 g 2.4 kg food) (fig.4F). Lf+ in combination with Phospholac 600™ has not led to the complete destruction of the tumor (as you can see from this document for the combination of Lf+ and milk fat). Rather, the overall effect was similar to the effect observed in monotherapy Phospholac 600™.

The combination of Lf+ and sphingomyelin was less effective than the corresponding types of monotherapy, such that tumor growth was the same as in control mice, suggesting that the two biologically active products can counteract each other (fig.4F).

Inclusion in the diet of mice c-9, t-11 CLA isomers at a rate of 5 g per 2.4 kg of diet, providing 3% of the fat component of the diet, only weakly inhibited the growth of 4T1 tumors and showed synergistic effects with Lf+.

Example 8

The example shows that milk fat inhibits tumor growth breast cancer line 4T1, both individually and in combination with chemotherapy.

A group of 6 mice of Balb/c mice were kept on a control diet AIN-93 or the same diet with substitution at the 5% milk fat, providing ~71% of the fat component of the diet. Two weeks after the diet mice were inoculated subcutaneously with 2×104tumor cells T. Mice were monitored tumor growth, and tumor size was measured every three days.

Compared with mice that were kept on a control diet, tumors in mice, which contained n the diet from milk fat, was produtivos three days later and at 35 days after inoculation, when mice slew formed tumors were smaller on average by 21% (P<0,05) (figure 5). Chemotherapeutic drug paclitaxel was administered administered intraperitoneally (30 mg/kg body weight), once tumors reached ~0.5 cm in diameter, which resulted in a reduction in the size of the tumors (35 day) to 31% (P<0,05) in mice, which were kept on a control diet. At 35 days, as a result of treatment with paclitaxel, in mice fed milk fat, the sizes of tumors were reduced by 35% (P<0.05) and 49% (P<0.01) compared with mice that were kept on a control diet with treatment with paclitaxel and mice, which were kept on a control diet without treatment, respectively.

Example 9

The example shows that milk fat inhibits the growth of tumours line 4T1 breast cancer, which disseminat in the lungs and liver.

The suppression of metastases in the lung: cell line 4T1 breast cancer have a high potential to metastasize and disseminatum in the lungs and liver. The lungs of mice in example 8 (day 35) were tested for the presence of tumors and metastases. The average number of tumors on the surface of the lungs of mice that were kept on a control diet without treatment, mice that were kept on a control diet with treatment with paclitaxel, we the her fed milk fat without treatment, and mice fed milk fat by treatment with paclitaxel, was 32, 18, 22 and 10, respectively (Fig.6).

Compared with mice that were kept on a control diet without treatment, treatment with paclitaxel and feeding milk fat at high doses was significantly reduced (P<0.01) number of tumors on the lung surface 44% and 31%, respectively.

Diet from milk fat in combination with treatment with paclitaxel resulted in a further decrease in the number of tumors, 63% (P<0.001) and 44% (P<0,05)compared with mice that were kept on a control diet without treatment with paclitaxel, and mice, which were kept on a control diet with treatment with paclitaxel, respectively. Accordingly, the weight of the lungs of mice that were kept on a control diet with treatment with paclitaxel, mice fed milk fat without treatment, and mice fed milk fat by treatment with paclitaxel, was significantly reduced by 30, 22, and 40% compared with mice that were kept on a control diet without treatment (table 7).

Table 7
Body weight, tumor, organ and tissue and the number of blood cells1
The mouse with the Pujol Mice without tumor
The control diet (n=6)Control diet + paclitaxel (n=6)Diet from milk fat (n=6)Diet from milk fat + paclitaxel (n=6)The control diet (n=6)Diet from milk fat (n=6)
Body weight (g)18,7±2,7217,7±1,520,3±2,020,8±1,4520,8±0,9821,2±2,3
Slaughter body weight (g)18,4±2,8217,6±1,420,1±1,9420,7±1,35--
Tumor (mg)263,5±31,7103,4±24,13143±20,9448,3±11,24--
Spleen (mg)79,2±9,472,35±8,23 82,34±7,5480,28±4,9582,6±12,782,3±9,5
Liver (mg)1268,9±86,31210,1±72,41230,8±63,71201,3±82,951204,5±102,51221,2±94,3
Lungs (mg)389,8±24,62274,5±30,63302,1±9,74234,5±25,65183,4±13,9190,1±17,8
Gastrocnemius muscle (mg)the 96.3±8,62to 101.2±9,3128,5±7,94to 129.2±10,85158,7±25,2171,7±12,52
Epididymal adipose tissue (mg)26,7±8,2224,3±6,755,3±7,1458,6±6,05154,4±23,9172,5±31,82
WBC (× 103/µl)4,21±0,572 2,37±0,393of 5.34±0,7645,48±0,565of 5.89±0,746,12±0,93
RBC (× 106/µl)6,35±0,6223,21±0,7438,65±0,934of 7.36±0,8758,23±0,588,67±0,62
1Data are expressed as mean ±SEM. Statistical significance was determined by student's t test.2P<0.05 compared with the body weight of mice without tumor, which contained on the control diet;3,4P<0.05 compared with the body weight of mice having tumor, which contained on the control diet;5P<0.05 compared with the body weight of mice having tumor, which contained on the control diet and treated with paclitaxel.

The suppression of metastases in the liver: similarly examined for the presence of tumors and metastases to the liver in mice above. The liver is cut out, made the cut and were stained with haematoxylin/eosin and counted the number of metastatic nodules within the liver. The average number of metastases in mice, which were kept on a control diet without treatment, the mice that contained on the completed dietary treatment with paclitaxel, in mice fed milk fat without treatment, and in mice fed milk fat by treatment with paclitaxel was 108, 59, 74 and 36, respectively (Fig.7). Thus, treatment with paclitaxel and feeding milk fat at high doses significantly (P<0,01) reduced the number of tumors in the liver by 45% and 32%, respectively, compared to mice, which were kept on a control diet without treatment.

Diet from milk fat in combination with treatment with paclitaxel resulted in a further decrease in the number of tumors by 67% (P<0.001) and 39% (P<0.05) as compared with mice that were kept on a control diet without treatment, and the mice that were kept on a control diet with treatment with paclitaxel, respectively.

Example 10

The example shows that milk fat inhibits tumor angiogenesis.

In order to assess tumor angiogenesis above the primary tumor 4T1 (example 8; 35 days) cut in mice, which were kept on a control diet without treatment, the mice were kept on a control diet with treatment with paclitaxel in mice fed milk fat without treatment, and in mice, which were kept on the diet from milk fat by treatment with paclitaxel, made the cut and were stained with antibody against CD31 to identify vascular endothelial cells. Blood vessels stained with mAb against CD31, is read in random areas selected blindly for the registration of the average density of blood vessels (8A), or the average distance to the nearest blood vessel, labeled mAb to CD31, from a point location (8B).

During treatment with paclitaxel and feeding milk fat at high doses significantly (P<0,05) decreased the density of microvessels (CD31+ vessels) by 37% and 31%, respectively, compared to mice, which were kept on a control diet without treatment (figa). Accordingly, when treated with paclitaxel and feeding milk fat at high doses significantly (P<0,05) increased the average distance to the nearest CD31+ vessels 71 and 59%, respectively (pigv).

Diet from milk fat in combination with treatment with paclitaxel resulted in a further decrease in the density of microvessels by 52% (P<0.001) and 22% (P<0.05) as compared with mice that were kept on a control diet without treatment, and the mice that were kept on a control diet with treatment with paclitaxel, respectively. Accordingly, the diet from milk fat in combination with treatment with paclitaxel led to a further increase in the average distance to the nearest CD31+ vessels to 125% (P<0.001) and 31% (P<0.05) as compared with mice that were kept on a control diet without treatment, and the mice that were kept on a control diet with treatment with paclitaxel, respectively.

Example 11

Cachexia is seriously the second problem for the subjects with malignant tumor, because it weakens the physical condition and reduces the response of subjects to the treatment. Improved nutrition is one of the ways to combat cachexia. In this experiment investigated, will improve if feeding milk fat condition of mice with cachexia model of breast cancer, used by the applicant, and will long-term feeding of high doses of milk fat, which contains saturated fatty acids (e.g., stearate), hypercholesterinemia, which would be unfavorable for some organs such as the liver and spleen. This example shows that a diet with milk fat at high doses is no apparent toxic effects on organs and weakens cachexia caused progressive malignant tumor.

The absence of toxic effects on organs. In the spleen and liver of mice that were fed high doses of milk fat, not observed obvious signs of toxic effects, and no significant changes in the mass of bodies (P>0.05) as compared with mice that were kept on control diet (table 7).

Reducing the symptoms of cachexia. Model 4T1 metastatic breast cancer represents an ideal model of cachexia in cancer. The inoculation of tumors led to the significant reduction in slaughter weight by 12% (P<0,05)that accounted for the lo the loss in mass of the gastrocnemius muscle and epididymal adipose tissue by 40% (P< 0.05) and 83% (P<0,01), respectively, compared to mice without tumors, which are contained on the same control diet (table 7). Reduced symptoms of cachexia was observed in mice that were kept on a diet with milk fat, as evidenced by significantly increased slaughter weight (P<0,05) (9,2 and 17.6%) and the mass of the gastrocnemius muscle (33.4 and 27.6 per cent) and epididymal adipose tissue (107 and 141%) in mice fed milk fat without treatment, and in mice fed milk fat by treatment with paclitaxel compared to those mice that were kept on a control diet without treatment, and mice, which contained on the control diet with treatment with paclitaxel, respectively. The increase in the mass of the gastrocnemius muscle and epididymal adipose tissue was not limited to mice-carriers of tumour, because feeding milk fat healthy mice without tumor, increased a lot the last two tissues 8.2 and 12% (P<0,05), respectively, compared to those of healthy mice without tumor, which contained on the control diet.

Example 12

One of the most severe side effects of the tumor and the treatment of malignant tumors, such as chemotherapy, is hematologic suppression, which can lead to immunologic failure, and that, in turn, can I have determined the cause of severe infections and even death. This example shows that a diet with milk fat decreases hematologic suppression caused by malignant tumor and chemotherapy.

Violation of haematopoiesis was found in mice with cachexia from example 11, above, because the number of erythrocytes (RBC) and leukocytes (WBC) was significantly reduced by 23 and 29% (P<0,05), respectively, compared to those of healthy mice without tumor, which contained the same control diet (table 7). Chemotherapy with paclitaxel resulted in a further reduction in the number of RBC and WBC 49 and 44% and a corresponding decrease in weight of the spleen by 8.5% compared with mice-carriers of tumour, which is contained on the control diet.

In contrast, the number of RBC and WBC in mice fed milk fat by treatment with paclitaxel, declined only 11% and 7% and the weight of the spleen at 3%, respectively, compared to those of healthy mice without tumor, which contained on the control diet (table 7). The number of WBC in mice-carriers of tumour fed milk fat without treatment, decreased only by 5%, while the number of RBC was slightly increased by 9% (P<0.05) and weight of the spleen was completely restored. Was significantly increased number of RBC and WBC in mice fed milk fat by treatment with paclitaxel, 56, and 57% (P<0,01) sootvetstvenno and 11% the mass of the spleen (P< 0.05) as compared to those mice that were kept on a control diet with treatment with paclitaxel (table 7). The number of RBC and WBC in mice-carriers of tumour fed milk fat without treatment, was increased by 34% and 27% (P<0.05) as compared to those mice-carriers of tumour, which is contained in a control diet without treatment.

Thus, the diet from milk fat weakens hematologic suppression by enhancing or restoring the number of erythrocytes and leukocytes, which reduced due to a malignant tumor and as a result of chemotherapy.

Example 13

Cytotoxic drugs damage the villi of the intestine, which leads to the fact that they are smooth, and thus, change the suction properties of the digestive tract (Melichar et al., 2005). This example shows that a diet with milk fat improves the condition caused by damage to the intestine caused by chemotherapy.

To determine defended whether milk fat digestive tract from damage caused by paclitaxel, a group of 6 mice of Balb/c mice were kept on a control diet AIN-93 or the same diet with substitution at the 5% milk fat, providing ~71% of the fat component of the diet. Two weeks later, each mouse was made intraperitoneally injection of paclitaxel (30 mg/kg body weight). Even after one week is ISA cut and separated skinny intestine. Immersed in the paraffin sections of the jejunum of mice that were kept on a control diet, a diet with milk fat control diet plus paclitaxel and diet from milk fat plus paclitaxel, were stained with hematoxylin and eosin.

As expected, healthy mice, which without treatment contained on the control diet and the diet with milk fat, had intact villi of the intestine. In contrast, the slices obtained from mice that were treated with paclitaxel, showed that the jejunum these mice contains damaged and flattened villi. Villi in mice fed milk fat by treatment with paclitaxel, were mostly intact, suggesting that the diet from milk fat has a protective effect on the mucous membrane of the intestine.

In order to determine the degree of damage to the intestine, we measured the average length of the fibers. Compared with villi control mice, paclitaxel significantly reduced the average length of fibers 61% (P<0,01) (figa). Diet from milk fat defended the intestines from damage caused by paclitaxel, since the average length of the fibers in mice fed milk fat by treatment with paclitaxel, was reduced by 29% (P<0.05) as compared with the control mice. Villi mice, which were kept on a diet with milk fat with treatment paclitaxel is, were significantly longer (40%, P<0.05)than the fibers of mice that were kept on a control diet with treatment with paclitaxel.

The activity of γ-glutamyltranspeptidase (γ-GGT) jejunum, the indicator of the brush the edges of the epithelium of the small intestine (Tate and Meister, 1981 and Ferraris, et al., 1992), was registered as a separate parameter injury of the intestine. Treatment with paclitaxel significantly reduced the level of γ-GGT in the mucosa of the jejunum by 56% (P<0.01) compared with control mice without treatment (pigv), whereas the activity of γ-GGT mice, which were kept on the diet from milk fat by treatment with paclitaxel was reduced only by 29% (P<0,05). The activity of γ-GGT jejunum was significantly increased in mice that were kept on the diet from milk fat by treatment with paclitaxel (78%, P<0.05) as compared to those mice that were kept on a control diet with treatment with paclitaxel.

Mucositis of the small intestine caused by chemotherapy, is associated with cell apoptosis in the crypts, which precedes atrophy of the villi (Keefe, et al., 2000). Very few apoptotic cells was in the crypts in mice, which without treatment contained on the control diet and the diet with milk fat. In contrast, sections of the intestine of mice that were kept on a control diet and treated with paclitaxel, contained a large number of cells in apoptosis. A diet supposedly cnym fat significantly reduced the number of apoptotic cells. Apoptotic cells were counted in 10 randomly selected crypts and were expressed as apoptotic Taurus on the crypt (figure 10). Treatment with paclitaxel significantly increased the number of apoptotic cells in 6-fold compared with mice without treatment. Diet from milk fat was significantly reduced the number of apoptotic cells in mice treated with paclitaxel by 39% (P<0.05) as compared with the control diet.

Discussion of examples 1-13

In the above-described experiments have shown that oral input a normal anhydrous milk fat and antitumor activity. Introduction milk fat increased the effect of immunotherapy on verified lymphoma and increased the effect of chemotherapeutic treatment verified tumors in breast cancer.

Milk fat in combination with Lf+ completely inhibited opukholeobrazovanie all subjects with lymphoma, leading to increased antitumor cytolytic activity of tumor apoptosis, and reduced the number of blood vessels of the tumor.

Introduction milk fat in combination with chemotherapeutic treatment slowed the growth of tumors in breast cancer, in addition, milk fat, or individually or in combination with chemotherapy treatment in breast cancer was significantly suppressed the metastasis of tumors in eggie and liver and the growth in these organs.

Antitumor activity was also observed when introducing fractions of milk fat. So, Phospholac 600™, and sphingomyelin resulted in a delay of tumor formation.

In addition, the introduction of milk fat reduces hematologic suppression, cachexia and damage to the intestine caused by tumors in breast cancer and/or chemotherapy.

Example 14

The example shows that milk fat significantly inhibits the mass loss due to chemotherapy and may contribute to weight gain.

Diet

Experimental food rations received by Crop & Food Research, Palmerston North, New Zealand, using as a control diet powdered drug AIN93G. In the AIN93G diet as a protein source used casein, and as a source of lipids used soybean oil. Soybean oil was replaced in the experimental food rations on anhydrous milk fat (AMF) so that the total content of lipids power remained unchanged. Diets included or 100, 35,7 or 7.1 g of AMF on 2 kg of food, providing 70, 25 and 5% of the total content of lipids in the diet (140 g lipid/2 kg). Fresh food was provided twice a week, and mice had free access to food and water throughout the study.

Experimental model for the analysis of side effects of chemotherapy

Cyclophosphamide enter the if in accordance with Animal Ethics Approval (University of Auckland). All experiments consisted of 6 mice in the treatment group, if not stated otherwise. Cyclophosphamide was dissolved in PBS, were injected I.P. Pavlova. in the amount of 300 mg/kg During the autopsy collected blood by cardiac puncture.

A group of six C57BL/6 contained on the AIN93G diet or the same diet with the replacement or 5%, 25%, or 70% milk fat and after 4 weeks of feeding at day 0 was administered intraperitoneally injected cyclophosphamide (300 mg/kg body weight). Mice were killed after 4, 8 and 12 days and analyzed the tissue for side effects treatment drug, as described below.

As shown in figa, day 0 was not significant differences in the mass of bodies mice (i.e. after 4 weeks of content on four different diets before chemotherapy). In contrast to this, namely through 4 days of chemotherapy, mice, which were kept on a control diet lost 7% body weight (pigv). In the case of two high doses of milk fat the degree of weight loss was significantly reduced by 54% at 4 days (P<0.001) and 39% (P=0,0017) respectively compared with the control diet. In the case of low doses of milk fat weight loss unreliable decreased by 19% (P=0,068) compared with the control. In mice on the control diet was observed a slight increase in weight gain after 12 days of chemotherapy, whereas, in contrast, the mass of the bodies of the mice, which is what's fed the lowest dose of milk fat, restored almost to normal (P<0,001), and mouse on a diet with the two higher doses of milk fat has actually gained weight (figs).

Example 15

The example shows that milk fat facilitates the recovery of white blood cells circulating in the blood, and spleen leukocytes (WBC) after chemotherapy, stimulates the development of colony forming units in the spleen and ensures the recovery of the spleen.

The content on different diets with milk fat within 4 weeks before starting chemotherapy did not influence significantly on the number of peripheral WBC. Cyclophosphamide significantly reduced the number of peripheral WBC in mice in example 14 to 4 days, more than 90%, regardless of diet (Fig). All four groups of mice, the number of WBC started to recover for 8 days. For 12 hours was significantly increased level of recovery in the number of WBC in the group of mice that were fed the highest dose of milk fat (P=0,0028). The number of WBC in these mice returned to normal. In contrast, lower doses of milk fat did not contribute significantly to the recovery in the number of WBC.

The content on different diets with milk fat within 4 weeks before starting chemotherapy did not influence significantly on the volume of the cellular content of the spleen (figa). Cyclophosphamide significantly reduced the volume of the cell content is letenky over 4 days in mice which contained the control diet. Compared with the control diet, the 4 day diet with 70%, 25% and 5% milk fat weakened the loss of spleen cell content of 52%, 41% and 25%, respectively. 8 day diet with 70% and 25% milk fat weakened the loss of spleen cell content of 32% and 27% respectively, while the 12 day diet with 70% milk fat had an impact on the volume of the cellular content of the spleen (figa).

Recovery of the number of cells within the spleen began with the formation of colony forming units in the spleen 4 days after starting chemotherapy. On the 8th day of each diet with 70%, 25% and 5% milk fat significantly stimulated the formation of colony-forming units in the spleen by 207%, 130% and 85%, respectively (pigv). For 12 hours the volume of the cellular content of the spleen in mice, which were kept on a diet with 70% and 25% milk fat, has almost recovered to normal values, thus it is not surprising that when the content in the diet from milk fat, the number of colony forming units was significantly decreased, whereas the spleen of mice that were kept on a control diet, were still subjected to recovery (pigv).

Example 16

The example shows that milk fat increases the size and hemoglobin content in erythrocytes (RBC) after chemotherapy.

Cyclophosphamide, an is independent of the power for 8 days reduced the number of peripheral RBC in mice from example 14 (figa). The loss of the number of RBC, which was less than the reduction in the number of WBC in example 15, ranged from 17% to 22% over 4 days. The number of RBC continued to decline to 8 days and 12 days not fully recovered. Diet from milk fat were not affected significantly by the reduction in the number of RBC-mediated cyclophosphamide. Despite the fact that at higher doses of milk fat was observed enhancement of recovery from the PNC, the results were insignificant.

It was unexpectedly found that each of the two higher doses of milk fat prevented the reduction and/or contributed to the restoration of the level of hematocrit (HCT) (pigv). The hematocrit is the percentage of whole blood that contains red blood cells, and is a way of measuring how the number of RBC and RBC size. Given that the diet from milk fat were not affected significantly by the number of RBC, it can be assumed that increased levels of HCT is a result of the increase in the average volume of RBC. Diet with 70% of milk fat was significantly (P=0,023) delayed reduction in the level of HCT for 8 days (a decrease of 18% compared to 31% reduction in the control diet) and increased the level of HCT almost to normal levels at 12 hours (P<0,001). Diet with 25% milk fat significantly contributed to the recovery of HCT levels at 8 (P=0,017) and 12 days (P=0,06), when on the 8th day observed 18% decrease compared to 31% reduction in the control diet and for 12 hours was observed in 9% decrease compared to the 24% decrease for the control diet. The results in the case of very low doses of milk fat did not reach significant values.

The key issue was the question of impact does the increase in the level of HCT is also on the increase in the total level of hemoglobin. Diet with 70% milk fat reduced reduced hemoglobin for 8 days, when there was a 22% decrease compared to 31% reduction in the control diet, but the difference did not reach significant values (figs). However, on the 12th day of this diet significantly increased the hemoglobin level of 8% (P=0,0048) compared with mice that were kept on a control diet. Diet with 25% milk fat significantly contributed to the restoration of hemoglobin level 8 (P=0,030) and 12 days (P=0,015), when the 8 days was observed a 20% decrease compared to 31% reduction in the control diet and 12 day observed 12% decrease compared with 26% decrease in the control diet. The results in the case of very low doses of milk fat did not reach significant values.

Thus, the results show that although the use of milk fat is not affected significantly by the number of RBC, it increases the size of RBC and hemoglobin in RBC and may, therefore, be who is one positive effect of anemia. These results have important implications for the treatment of iron deficiency anemia, for example, microcytic anemia, which is characterized pathologically small size of the RBC.

Example 17

As described above, mucositis occurs when chemotherapy of malignant tumors destroys rapidly dividing epithelial cells lining the gastrointestinal tract, leaving the mucous membrane tissues open to ulceration and infection. The example shows that milk fat reduces the damage of the intestine due to chemotherapy.

To determine defended whether milk fat gastro-intestinal tract from damage caused by the action of cyclophosphamide, skinny intestine of mice in example 14 was divided into slices, immersed in paraffin and stained with hematoxylin and eosin.

Feeding for 4 weeks before chemotherapy different diets with milk fat had no apparent effect on villi of jejunum (Fig). Sections of the intestine of mice that did one injection of cyclophosphamide showed that the jejunum of mice contained destroyed and flattened villi. To assess the damage caused by chemotherapy, used an average length of fibers. Cyclophosphamide was significantly reduced average length of fibers by 65% (P<0,001) compared to healthy control mice (Fig). Diet with 70%, 25% and 5% milk fat was protected from the damage of the tion of the intestine, caused by cyclophosphamide, since the average length of the fibers in mice, which were kept on the diet from milk fat by treatment with cyclophosphamide, 4 days decreased by 36% (P<0.01), and 41% (P<0.01) and 52% (P<0,001), respectively, and for 8 days by 21% (P<0.05)and 24% (P<0.01) and 31% (P<0,01), respectively, compared to mice, which were kept on the control diet. Villi after chemotherapy in mice, which were kept on a diet with 70% and 25% milk fat by treatment with cyclophosphamide, were significantly longer than 4 days (71% and 58%, respectively, for both diets P<0.01) and 8 days (44% and 36% respectively for both diets P<0.05)than in mice, which were kept on a control diet with treatment with cyclophosphamide. On the 12th day of the intestine of mice that were kept on a control diet began to recover, and only the highest dose of milk fat showed the preservation of the authentic effect.

The results suggest that diets with milk fat have a protective effect on the intestinal mucosa, especially during the first few days after beginning chemotherapy, when side effects from treatment are the most obvious. Not wanting to be limited by any theory, suggest that the protection of the intestine through nutrition from milk fat at least partially leads to a reduced mass loss due to chemotherapy and p is increased to increase the mass, observed in mice fed these foods. Diet from milk fat, apparently, helps to maintain appetite and absorption properties of the gastrointestinal tract. The weight increase was not only due to the high caloric content of milk fat, as the diet is balanced by the intensity and mice fed milk fat, not gaining weight during the four-week feeding period before chemotherapy.

Discussion of examples 14-17

In examples 1-13, described above, investigated the effect of milk fat on side effects from chemotherapy in mice-carriers of tumour. In examples 14-17 were investigated the direct effect of milk fat on side effects, mediated by chemotherapy with cyclophosphamide, other control healthy mice. Cyclophosphamide is a drug that strongly inhibits the metabolism and cell division, thus, as a side effect of cyclophosphamide inhibits the growth and division of cells of the immune and circulatory systems, leading to mielosupression and anemia. In the research reported that anemia caused by chemotherapy, including anemia mild to moderate, has a negative impact on the quality of life of subjects with malignant tumor (Groopman JE, Itri LM. Chemotherapy-induced anemia in adults: incidence and treatment. J Natl Cancer Inst. 1999 Oct 6;91 (19):1616-34). Chemotherapy can also reduce the quality of life of subjects with malignant tumor, determining the mass loss due to poor appetite, mucositis, nausea, damage to the bowel and dehydration.

The results obtained herein show that milk fat is effective in preventing or reducing the side effects of chemotherapy.

Example 18

The example shows that milk fat acts synergistically with Lf+ to reduce many of the side effects of chemotherapy in a subject, including hematologic suppression, anemia and damage to the gastrointestinal tract.

Diet

Experimental food rations received from the Crop & Food Research, Palmerston North, New Zealand, using as a base powder composition AIN93G. In the AIN93G diet used casein as the protein source and soybean oil as a source of lipids. Soybean oil in experimental food rations were replaced by anhydrous milk fat (AMF) so that the total content of lipids in the diet did not change. The casein in the experimental food rations were replaced by Lf+, so that the total protein content in the diet was not changed. Diet with substitution at AMF contained 35 g of AMF on 2 kg of food, providing 25% of the total lipid content of the food (140 g lipid/2 kg). Diet with replacement at Lf+ contained or 0.1, 1 or 10 g Lf+ 2 kg of food, providing of 0.025, 0.5 and 2.5% of the total protein supply (400 g protein/2 kg). Some diets contain a combination of 35 g of AMF on 2 kg power supply and one of the above quantities Lf+. Fresh food was provided twice a week, and mice had free access to food and water throughout the study.

Experimental model for the analysis of side effects of chemotherapy

Cyclophosphamide was administered in accordance with Animal Ethics Approval (University of Auckland). In all experiments consisted of 24 mice per treatment group. Cyclophosphamide dissolved in PBS, were injected I.P. Pavlova. the rate of 300 mg/kg For blood collection mice were subjected to deep anesthesia and were bled by cardiac puncture.

A group of 24 mice C57BL/6, containing an equal number of female and male mice, contained on the AIN93G diet or the same diet with the substitution for milk fat, Lf+, or a combination of milk fat and Lf+ and after 4 weeks of feeding at day 0 was administered intraperitoneally injected cyclophosphamide (300 mg/kg body weight). Mice were killed by groups of six mice at day 0, 4, 8 and 12 days later, and analyzed blood and tissue of the intestine for the study of adverse effects of the medicinal product, as described below.

The average length of the fibers were analyzed for 8 days, using two-factor analysis of variance (ANOVA) to assess the effects of replacement of milk fat (0 vs. 25%), substitution Lf+ (0 vs. 0,025%) and their interaction. Separate groups were compared, applied the method of multiple comparisons Tukey's.

Milk fat acts synergistically with Lf+to inhibit damage to the intestine caused by chemotherapy. The content on different diets with milk fat within 4 weeks before chemotherapy had no significant influence on the length of the villi of the jejunum (Fig). The average length of fibers was used as an assessment of the damage caused by chemotherapy. Cyclophosphamide was significantly reduced average length of fibers in all groups at 4 days (Fig). 25% milk fat contributed to the recovery of the intestine from damage caused by cyclophosphamide. For 8 days the average length of the fibers in the group with milk fat was significantly greater (P<0,001)than in the control group.

The combination of milk fat and Lf+ resulted in significantly greater length of the villi compared only with milk fat and only with Lf+ (P<0,01).

Example 19

The example shows that milk fat acts synergistically with Lf+to contribute to the restoration of circulating leukocytes (WBC) after chemotherapy.

In order to evaluate the effects of enrichment of milk fat (0 vs. 25%), enrichment Lf+ (0 vs. 0,025%) and their combinations analyzed the difference in the number of WBC between 4 days and 8 days, using two-factor covariance analysis (ANCOVA). Body weight during chemotherapy was included as an independent variable. The combination of milk fat and Lf+ compared only with milk is fat or only Lf+, using one-sided criterion Dannetta.

The number of WBC in samples from cardiac puncture was recorded in days of chemotherapy and after 4, 8 and 12 days as described in example 18. The content in the diet from milk fat and diet with 0,025% Lf+ within 4 weeks before chemotherapy had no significant effect on the number of peripheral WBC (Fig). Cyclophosphamide significantly reduced the number of peripheral WBC over 4 days in all mice in example 19. The number of WBC started to recover for 8 days in all groups of mice. Therefore, compared the number of WBC in the case of the lowest values (4 days) 8 days, when the number of WBC already recovered substantially. Compared to the diet with 0,025% Lf+ (P=0,049) and diet from milk fat (P=0,012) the increase in the number of WBC was significantly greater in mice fed 0,025% Lf+ and milk fat in combination.

Example 20

The example shows that milk fat acts synergistically with Lf+to increase the number and size of red blood cells (RBC) after chemotherapy, with a tendency to increase in hemoglobin.

In order to evaluate the effects of enrichment of milk fat (0 vs. 25%), enrichment Lf+ (0 vs. 0,025%) and their combinations were analyzed, the number of RBC, HCT, and hemoglobin using two-factor analysis (ANCOVA). Body weight during chemotherapy was included as an independent variable. The combination of milk fat is Lf+ compared only with milk fat or only Lf+, using one-sided criterion Dannetta.

The number of RBC, HCT and HGB levels in samples cardiac puncture was recorded in days of chemotherapy and after 4, 8 and 12 days as described in example 18.

Cyclophosphamide reduced the number of RBC at 4 and 8 days in mice in example 20, regardless of diet (Fig). The number of RBC continued to decline to 8 days and then began to recover, but has not reached full recovery for 12 hours. Milk fat showed a drop at 4 and 8 days and acted synergistically with 0.25% Lf, which helped to restore the number of RBC for 12 hours. The number of RBC for 12 hours was significantly greater in mice fed the 0.25% Lf+ and milk fat in combination compared with a diet with 0.25% Lf+ (P=0,018) and diet from milk fat (P=0,024).

Cyclophosphamide was reduced HCT at 4 and 8 days in mice in example 20, regardless of diet (Fig). HCT has continued to decline to 8 days and then began to recover, but has not reached full recovery for 12 hours. Milk fat showed a decline falling 4 and 8 days and acted synergistically with 0.25% Lf, which contributed to the recovery of HCT levels for 12 hours. The HCT levels at 12 hours were significantly greater in mice fed the 0.25% Lf+ and milk fat in combination compared with a diet with 0.25% Lf+ (P=0,046) and diet from milk fat P=0,047).

Cyclophosphamide reduced the levels of hemoglo the ina at 4 and 8 days in mice in example 20, regardless of diet (Fig). Hemoglobin continued to decrease to 8 days and then began to recover, but did not achieve full recovery for 12 hours. Milk fat showed a decline falling 4 and 8 days and acted synergistically with 0.25% Lf, which contributed to the restoration of hemoglobin level for 12 hours. The hemoglobin level for 12 hours was significantly higher in mice fed the 0.25% Lf+ and milk fat in combination compared with a diet with 0.25% Lf+ (P=0.038). And was higher but not significantly, compared with the diet from milk fat (P=0,094).

Thus, the results show that the use of milk fat in combination with 0.25% Lf+ significantly increases the number and size of RBC, and there is a tendency to increase in the total hemoglobin content. The results have important implications for the treatment of anemia, including anemia caused by chemotherapy.

Example 21

The example shows that milk fat significantly reduces anorexia and cachexia, which are caused by chemotherapy. Diets containing a combination of milk fat and Lf+reduce anorexia and cachexia, which are caused by chemotherapy.

Body weight was recorded daily starting chemotherapy and after 4, 8 and 12 days as described in example 18. Milk fat was significantly (P=0.004) increased the mass of bodies mice regardless of gender, enrichment Lf+ and days, i.e. due to the loss of any significant inter is the behaviour, combined analysis of animals of both sexes, all day and all groups feeding with Lf (Fig). The effects of milk fat were specific to chemotherapy, since neither diet with milk fat or other diets did not have any reliable effect on the mass of the bodies of the mice before chemotherapy.

Industrial application

Methods, medical applications and compositions of the present invention are useful in the inhibition of tumor growth, maintaining or improving one or more parameters, such as the number of white blood cells, red blood cells or myeloid cells, while stimulating the immune system and in the treatment or prevention of a malignant tumor. Methods and therapeutic use can be implemented through the use of diet (as food or food additives to the food product), nutraceutical or pharmaceutical compositions.

Specialists in this field will understand that the above description is provided only as a way of illustration and that the invention is not limited to them.

Links

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1. A method of treating or preventing mucositis, anemia caused by low levels of hemoglobin or red blood cells, cachexia, radiation, or maintain, or improve one or more parameters, such as the number of leukocytes, red blood cells and myeloid cells in a subject, comprising introducing an effective amount of milk fat or similar milk fat needy in this subject.

2. The method according to claim 1, further comprising separate, simultaneous or sequential administration to the subject one or more anticancer agents.

3. The method according to claim 1 or 2, where one or more anticancer agents selected from the group comprising lactoferrin, APO-lactoferrin, lactoferrin polypeptide, a functional lactoferrin variant, a functional fragment of lactoferrin,lactoferrin, containing metal ion, natural saturated iron lactoferrin, lactoferrin, essentially fully saturated with iron, a functional lactoferrin variant containing a metal ion, a functional fragment of lactoferrin containing metal ion, or a combination of two or more of them, antitumor factors nutritional products including vitamin D (including vitamin D1 [luisteren], vitamin D2 [calciferol or ergocalciferol], vitamin D3 [cholecalciferol], vitamin D4 [22-dihydroergosterol] and vitamin D5 [situaltional] and vitamin D5 [7-dehydrocholesterol]), analogues of vitamin D, soy protein, one or more components of soybeans, one or more omega-3 fatty acids from soy, one or more isoflavones from soy, genistein, daidzein, one or more peptides lunasin, one or more polyphenols, lycopene, wheat bran, flavonoids, Inositol, resveratrol, propolis, extracts of mushrooms, anthocyanins, almond, ginseng, casein hydrolysate, and combinations of two or more of them, chemotherapeutic drugs, immunotherapy, haematopoietic funds protivorahiticescoe funds or protivopokazanie tools and combinations of two or more of them.

4. The method according to claim 1 or 2, where the milk fat selected from the group comprising milk lipids, fractions of milk lipids, hydrolysates of milk lipids, practicerelated of milk lipids, fat cow's milk, cream, butter, anhydrous milk fat, buttermilk, whey oils, solid fractions of milk fat, soft fractions of milk fat fractions sphingomyelin, faction shells beads of milk fat, phospholipid fractions, and complex lipid fractions, and combinations thereof, and hydrolysates and fractions of the hydrolysates, and combinations of the hydrolysates and/or non-hydrolyzed fraction.

5. The method according to claim 1 or 2, where the milk fat or similar milk fat is an anhydrous milk fat or cream or, where the milk fat or similar milk fat includes:
(a) from 23% (wt./mass.) up to 32% (wt./mass.) palmitic acid;
(b) from 15% (wt./mass.) up to 22% (wt./mass.) oleic acid;
(c) from 10% (wt./mass.) up to 15% (wt./mass.) stearic acid;
(d) from 9% (wt./mass.) up to 12% (wt./mass.) myristic acid;
(e) from 3% (wt./mass.) up to 5% (wt./mass.) butyric acid;
(f) any two of a), b), C), d) or e);
(g) any three of a), b), C), d) or e);
(h) any four of a), b), C), d) or e); or
(i) each of a), b), C), (d) and (e).

6. The use of milk fat or similar milk fat for the treatment of mucositis, anemia, mediated by low hemoglobin or red blood cells, cachexia, mucositis or leukopenia, or maintain, or improve one or more indicators of the number of cells, number of erythrocytes and the number myeloid the x cells in the subject.

7. The use according to claim 6, where the milk fat or similar milk fat formulated for separate, simultaneous or sequential injection with one or more anticancer agents.

8. The use according to claim 6, where the milk fat or similar milk fat present in the composition, optionally comprising one or more anticancer agents.

9. The use according to any one of claims 7 or 8, wherein the one or more anticancer drugs selected from the group comprising lactoferrin, APO-lactoferrin, lactoferrin polypeptide, a functional lactoferrin variant, a functional fragment of lactoferrin, lactoferrin containing metal ion, natural saturated iron lactoferrin, lactoferrin, essentially fully saturated with iron, a functional lactoferrin variant containing a metal ion, a functional fragment of lactoferrin containing metal ion, or a combination of two or more of them, antitumor factors nutritional products including vitamin D (including vitamin D1 [luisteren], vitamin D2 [calciferol or ergocalciferol], vitamin D3 [cholecalciferol], vitamin D4 [22-dihydroergosterol] and vitamin D5 [situaltional] and vitamin D5 [7-dehydrocholesterol]), analogues of vitamin D, soy protein, one or more components of soybeans, one or more omega-3 fatty acids from soy, and one the or more isoflavones from soy genistein, daidzein, one or more peptides lunasin, one or more polyphenols, lycopene, wheat bran, flavonoids, Inositol, resveratrol, propolis, extracts of mushrooms, anthocyanins, almond, ginseng, casein hydrolysate, and combinations of two or more of them, chemotherapeutic drugs, immunotherapy, haematopoietic funds protivorahiticescoe funds or protivopokazanie tools and combinations of two or more of them.

10. The use according to any one of PP 8, wherein the milk fat is selected from the group comprising milk lipids, fractions of milk lipids, hydrolysates of milk lipids, fractions of the hydrolysates of milk lipids, fat cow's milk, cream, butter, anhydrous milk fat, buttermilk, whey oils, solid fractions of milk fat, soft fractions of milk fat fractions sphingomyelin, faction shells beads of milk fat, phospholipid fractions, and complex lipid fractions, and combinations thereof, and hydrolysates and fractions of the hydrolysates, and combinations of the hydrolysates and/or non-hydrolyzed fraction.

11. The use according to any one of PP 8, wherein the milk fat or similar milk fat is an anhydrous milk fat or cream or, where the milk fat or similar milk fat includes:
(a) from 23% (wt./mass.) up to 32% (wt./mass.) palmitic acid;
(b) from 15% (wt./mass.) up to 22% (massmass.) oleic acid;
(c) from 10% (wt./mass.) up to 15% (wt./mass.) stearic acid;
(d) from 9% (wt./mass.) up to 12% (wt./mass.) myristic acid;
(e) from 3% (wt./mass.) up to 5% (wt./mass.) butyric acid;
(f) any two of a), b), C), d) or e);
(g) any three of a), b), C), d) or e);
(h) any four of a), b), C), d) or e); or
(i) each of a), b), C), (d) and (e).

12. Composition for the treatment of mucositis or suitable for administration to a subject with a risk of suffering from, or subject to treatment for mucositis comprising from 2 grams to 210 grams of milk fat or similar milk fat and from 0.7 g to 210 g of one or more anticancer agents.

13. The composition according to item 12, comprising from 2 grams to 210 grams of milk fat or similar milk fat and from 0.7 g to 70 g of one or more anticancer agents.

14. The composition according to item 12, comprising from 35 grams to 210 grams of milk fat or similar milk fat and from 0.7 g to 210 g of one or more anticancer agents.

15. The composition according to item 12, comprising from 10 g to 200 g of milk fat or similar milk fat and 2.5 grams to 70 grams of one or more anticancer agents.

16. The composition according to item 12, comprising from 10 g to 200 g of milk fat or similar milk fat and from 0.7 g to 5 g of one or more anticancer agents.

17. The composition according to item 12, comprising from 15 g to 30 g of milk fat or similar milk fat and 1.0 g do,0 grams of one or more anticancer agents.

18. The composition according to item 12, comprising from 3 g to 8 g of milk fat or similar milk fat from 0.1 g to 1.0 g of one or more anticancer agents.

19. Composition according to any one of p-18, which represents an edible consumer product or part edible consumer product selected from the group comprising a confectionery product, a dessert, a liquid drink, briquetted food concentrate, cereal, spread, sauce, sauce, ice cream, yogurt, cheese, milk shakes, yogurt drink, milk powder, baby food or food for young or enteral fluid.

20. Composition according to any one of p-18, where one or more anticancer drugs selected from the group comprising lactoferrin, APO-lactoferrin, lactoferrin polypeptide, a functional lactoferrin variant, a functional fragment of lactoferrin, lactoferrin containing metal ion, natural saturated iron lactoferrin, lactoferrin, essentially fully saturated with iron, a functional lactoferrin variant containing a metal ion, a functional fragment of lactoferrin containing metal ion, or a combination of two or more of them, antitumor factors nutritional products including vitamin D (including vitamin D1 [luisteren], vitamin D2 [calciferol or ergocalciferol the l], vitamin D3 [cholecalciferol], vitamin D4 [22-dihydroergosterol] and vitamin D5 [situaltional] and vitamin D5 [7-dehydrocholesterol]), analogues of vitamin D, soy protein, one or more components of soybeans, one or more omega-3 fatty acids from soy, one or more isoflavones from soy, genistein, daidzein, one or more peptides lunasin, one or more polyphenols, lycopene, wheat bran, flavonoids, Inositol, resveratrol, propolis, extracts of mushrooms, anthocyanins, almond, ginseng, casein hydrolysate, and combinations of two or more of them, chemotherapeutic drugs, immunotherapy, haematopoietic funds protivorahiticescoe funds or protivopokazanie tools and combinations of two or more of them.

21. Composition according to any one of p-18, where the milk fat selected from the group comprising milk lipids, fractions of milk lipids, hydrolysates of milk lipids, fractions of the hydrolysates of milk lipids, fat cow's milk, cream, butter, anhydrous milk fat, buttermilk, whey butter, milk solids, ICRA, soft fractions of milk fat fractions sphingomyelin, faction shells beads of milk fat, phospholipid fractions, and complex lipid fractions, and combinations thereof, and hydrolysates and fractions of the hydrolysates, and combinations of the hydrolysates and/or non-hydrolyzed fraction.

22. Composition according to any one of p-18, where the milk fat or similar milk fat is an anhydrous milk fat or cream or, where the milk fat or similar milk fat includes:
(a) from 23% (wt./mass.) up to 32% (wt./mass.) palmitic acid;
(b) from 15% (wt./mass.) up to 22% (wt./mass.) oleic acid;
(c) from 10% (wt./mass.) up to 15% (wt./mass.) stearic acid;
(d) from 9% (wt./mass.) up to 12% (wt./mass.) myristic acid;
(e) from 3% (wt./mass.) up to 5% (wt./mass.) butyric acid;
(f) any two of a), b), C), d) or e);
(g) any three of a), b), C), d) or e);
(h) any four of a), b), C), d) or e); or
(i) each of a), b), C), (d) and (e).

23. Composition for the treatment of mucositis or suitable for administration to a subject with a risk of suffering from, or subject to treatment for mucositis comprising from 2 grams to 210 grams of milk fat or similar milk fat and from 0.1 g to 210 g of one or more anticancer agents.

24. The composition according to item 23, comprising from 0.1 g to 50 g of milk fat or similar milk fat and from 1.0 mg to 1.0 g of one or more anticancer agents.

25. The composition according to item 23, comprising from 0.1 g to 50 g of milk fat or similar milk fat Yot 1.0 mg to 1.0 mg of one or more anticancer agents.

26. The composition according to item 23, comprising from 2.0 g to 210 g of milk fat or similar milk fat and 0.7 mg on the 70,0 one or more anticancer agents.

27. The composition according to item 23, comprising from 35 grams to 210 grams of milk fat or similar milk fat and 0.35 g to 210 g of one or more anticancer agents.

28. The composition according to item 23, comprising from 10 g to 200 g of milk fat or similar milk fat and 2.5 g to 70.0 g of one or more anticancer agents.

29. The composition according to item 23, comprising from 10 g to 200 g of milk fat or similar milk fat and from 0.25 g to 5.0 g of one or more anticancer agents.

30. The composition according to item 23, comprising from 15 g to 30 g of milk fat or similar milk fat and from 1.0 g to 6.0 g of one or more anticancer agents.

31. The composition according to item 23, comprising from 3.0 g to 8.0 g of milk fat or similar milk fat and from 0.1 g to 1.0 g of one or more anticancer agents.

32. Composition according to any one of p-31, which represents an edible consumer product or part edible consumer product selected from the group comprising a confectionery product, a dessert, a liquid drink, briquetted food concentrate, cereal, spread, sauce, sauce, ice cream, yogurt, cheese, milk shakes, yogurt drink, milk powder, baby food or food for young or enteral fluid.

33. Composition according to any one of p-31, where the one or more protivoop is of evich drugs selected from the group including lactoferrin, APO-lactoferrin, lactoferrin polypeptide, a functional lactoferrin variant, a functional fragment of lactoferrin, lactoferrin containing metal ion, natural saturated iron lactoferrin, lactoferrin, essentially fully saturated with iron, a functional lactoferrin variant containing a metal ion, a functional fragment of lactoferrin containing metal ion, or a combination of two or more of them, antitumor factors nutritional products including vitamin D (including vitamin D1 [luisteren], vitamin D2 [calciferol or ergocalciferol], vitamin D3 [cholecalciferol], vitamin D4 [22-dihydroergosterol] and vitamin D5 [situaltional] and vitamin D5 [7-dehydrocholesterol]), analogues of vitamin D, soy protein, one or more components of soybeans, one or more omega-3 fatty acids from soy, one or more isoflavones from soy, genistein, daidzein, one or more peptides lunasin, one or more polyphenols, lycopene, wheat bran, flavonoids, Inositol, resveratrol, propolis, extracts of mushrooms, anthocyanins, almond, ginseng, casein hydrolysate, and combinations of two or more of them, chemotherapeutic drugs, immunotherapy, haematopoietic funds protivorahiticescoe funds or protivopokazanie tools and combinations of two or the more of them.

34. Composition according to any one of p-31, where the milk fat selected from the group comprising milk lipids, fractions of milk lipids, hydrolysates of milk lipids, fractions of the hydrolysates of milk lipids, fat cow's milk, cream, butter, anhydrous milk fat, buttermilk, whey oils, solid fractions of milk fat, soft fractions of milk fat fractions sphingomyelin, faction shells beads of milk fat, phospholipid fractions, and complex lipid fractions, and combinations thereof, and hydrolysates and fractions of the hydrolysates, and combinations of the hydrolysates and/or non-hydrolyzed fraction.

35. Composition according to any one of p-31, where the milk fat or similar milk fat includes:
(a) from 23% (wt./mass.) up to 32% (wt./mass.) palmitic acid;
(b) from 15% (wt./mass.) up to 22% (wt./mass.) oleic acid;
(c) from 10% (wt./mass.) up to 15% (wt./mass.) stearic acid;
(d) from 9% (wt./mass.) up to 12% (wt./mass.) myristic acid;
(e) from 3% (wt./mass.) up to 5% (wt./mass.) butyric acid;
(f) any two of a), b), C), d) or e);
(g) any three of a), b), C), d) or e);
(h) any four of a), g), C), d) or e); or
(i) each of a), b), C), (d) and (e).

36. Product for the treatment of mucositis or suitable for administration to a subject with a risk of suffering from, or subject to the treatment of mucositis, including milk fat or similar milk fat and one or more additional the additional anticancer agents, in the form of a combined preparation for simultaneous, separate or sequential use.

37. Product p, where an antitumor agent suitable for parenteral administration.

38. Product p or 37, where the one or more anticancer agents selected from the group comprising lactoferrin, APO-lactoferrin, lactoferrin polypeptide, a functional lactoferrin variant, a functional fragment of lactoferrin, lactoferrin containing metal ion, natural saturated iron lactoferrin, lactoferrin essentially fully saturated with iron, a functional lactoferrin variant containing a metal ion, a functional fragment of lactoferrin containing metal ion, or a combination of two or more of them, antitumor factors nutritional products including vitamin D (including vitamin D1 [luisteren], vitamin D2 [calciferol or ergocalciferol], vitamin D3 [cholecalciferol], vitamin D4 [22-dihydroergosterol] and vitamin D5 [situaltional] and vitamin D5 [7-dehydrocholesterol]), analogues of vitamin D, soy protein, one or more components of soybeans, one or more omega-3 fatty acids from soy, one or more isoflavones from soy, genistein, daidzein, one or more peptides lunasin, one or more polyphenols, lycopene, wheat bran, flavonoids, Inositol, resveratrol, propolis, extras the CT of mushrooms, anthocyanins, almond, ginseng, casein hydrolysate, and combinations of two or more of them, chemotherapeutic drugs, immunotherapy, haematopoietic funds protivorahiticescoe funds or protivopokazanie tools and combinations of two or more of them.

39. Product p or 37, where the milk fat selected from the group comprising milk lipids, fractions of milk lipids, hydrolysates of milk lipids, fractions of the hydrolysates of milk lipids, fat cow's milk, cream, butter, anhydrous milk fat, buttermilk, whey oils, solid fractions of milk fat, soft fractions of milk fat fractions sphingomyelin, faction shells beads of milk fat, phospholipid fractions, and complex lipid fractions, and combinations thereof, and hydrolysates and fractions of the hydrolysates, and combinations of the hydrolysates and/or non-hydrolyzed fraction.

40. Product p or 37, where the milk fat or similar milk fat includes:
(a) from 23% (wt./mass.) up to 32% (wt./mass.) palmitic acid;
(b) from 15% (wt./mass.) up to 22% (wt./mass.) oleic acid;
(c) from 10% (wt./mass.) up to 15% (wt./mass.) stearic acid;
(d) from 9% (wt./mass.) up to 12% (wt./mass.) myristic acid;
(e) from 3% (wt./mass.) up to 5% (wt./mass.) butyric acid;
(f) any two of a), b), C), d) or e);
(g) any three of a), b), C), d) or e);
(h) any four of a),g), C), (d) or (e); or
(i) each of a), b), C), (d) and (e).

41. The use of milk fat or similar milk fat for the manufacture of a medicinal product for administration to a subject with a risk of suffering from, or subject to the treatment of mucositis, anemia, caused by low hemoglobin or red blood cell count, cachexia, radiation, or maintain or improve one or more indications selected from the number of cells, number of erythrocytes and the number of myeloid cells in the subject.



 

Same patents:

FIELD: medicine.

SUBSTANCE: there are presented versions of the peptide (A) or (B) with the amino acid sequence of SEQ ID NO: 1 or 2 respectively presented in this description. The peptide has activity to induce a cytotoxic T-cell when an antigen-presenting cell carrying HLA-A2 (A*0201) presents it. There are described the versions of the peptide antibodies prepared by immunisation by the proper peptide. There are presented: an agent, methods for inducing: a cytotoxic (killer) T-cell, an antigen-presenting cell, CDH3 expressing cancer immunity; as well as a method of treating CDH3 expressing cancer on the basis of the peptide. What is presented is an isolated cytotoxic T-cell induced by said method on the basis of the peptide. There are described: the antigen-presenting cell and exosome presenting the complex containing the peptide and HLA-A2 (A*0201).

EFFECT: higher effectiveness of the use of the invention in treating CDH3 expressing cancer.

15 cl, 5 dwg, 2 tbl, 5 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: present invention refers to an aminopropylidene derivative presented by formula wherein R1 and R2, which may be identical or different, represent hydrogen or a substitute specified in the following (a)-(c), provided the case of both representing hydrogen is excluded: (a) carbonyl substituted with hydroxy, alkoxy or hydroxy alkylamino, (b) carbonylalkyl substituted by hydroxy or alkoxy, and (c) acrylic acid including its alkyl ester, R3 and R4, which may be identical or different, represent hydrogen, alkyl which may be substituted by phenyl or cycloalkyl, or R3 and R4, which together form a heterocyclic ring with a nitrogen atom bound thereto, represent pyrrolidino, piperidino, which may be substituted by oxo or piperidino, piperazinyl substituted by alkyl or penyl, morpholino or thiomorpholino; A means oxo or is absento, B represents canbon or oxygen; one of X and Y represents carbon, while the other one represents sulphur, a part represented by a dash line represents a single bond or a double bond, and a wavy line represents a cys-form and/or a transform. Also, the invention refers to a pharmaceutical composition exhibiting histamine receptor antagonist activity on the basis of said compounds.

EFFECT: there are produced new compounds and pharmaceutical compositions thereof, which can be used in medicine for treating asthma, allergic rhinitis, pollen allergy, hives and atopic dermatitis.

10 cl, 12 tbl, 58 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to medicine and concerns a pharmaceutical composition in the form of a capsule either acid-resistant, or with an acid-resistant coating containing recombinant interferon specified from alpha, beta, gamma recombinant interferon; a stabiliser of the biological, physical and chemical properties specified in low-molecular polyvinylpyrrolidone, macrogol 400-12000, propylene glycol, hydroxypropyl methylcellulose; a stabiliser of microbial contamination resistance specified in dexamethasone, diflorazone, boric acid, phospholipids, cyclodextrins, calcium stearate, and/or magnesium stearate, a thickly-forming base specified in hydrophilic, lipophilic, hydrophilic-lipophilic bases.

EFFECT: invention provides a higher therapeutic effect and prolonged shelf life.

3 cl, 4 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmacology, particularly to an agent with immunocorrective action on the humoral component of the immunity system in cytostatic immunosuppression. The agent with immunocorrective action on the humoral component of the immunity system in cytostatic immunosuppression, which is a liquid extract of licorice root (Radices Glycyrrhizae) prepared by repercolation with 40% ethanol in certain ratio materials: extractant.

EFFECT: licorice extract exhibits the immunocorrective properties expressed in the effective enhancement of the cyclophosphamide-suppressed humoral immune response.

1 tbl, 1 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmacology, particularly to an agent with immunocorrective action on the humoral component of the immunity system in cytostatic immunosuppression. The agent with immunocorrective action on the humoral component of the immunity system in cytostatic immunosuppression, which is a liquid extract of licorice root (Radices Glycyrrhizae) prepared by repercolation with 40% ethanol in certain ratio materials: extractant.

EFFECT: licorice extract exhibits the immunocorrective properties expressed in the effective enhancement of the cyclophosphamide-suppressed humoral immune response.

1 tbl, 1 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: present invention refers to pharmaceutical compositions containing synthetic oxidised lipids.

EFFECT: developing the method for using the oxidised lipids for treating and preventing the inflammation associated with the endogenous oxidised lipid.

39 cl, 11 tbl, 15 ex, 25 dwg

Antibodies // 2482131

FIELD: medicine, pharmaceutics.

SUBSTANCE: present invention refers to immunology. There are presented the antibodies that bind to the P-selectin glycoprotein ligand-1 (PSGL-1), as well as methods for stimulating the death of activated T-cells and simulating T-cell immune response in a patient by the use of the antibodies under the invention, and pharmaceutical compositions containing the antibodies under the invention. There are also disclosed nucleic acids, expression vectors and host cells for producing the antibodies under the invention.

EFFECT: invention may can find further application in therapy of the PSGL-1 associated diseases.

35 cl, 4 ex, 3 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to combinations of peptides in each case with the same sequence length (SEQL) which can be prepared in a stable reproducible quality and quantity of a mixture (A) containing a number of x amino acid with protected acid groups or a number of z peptides with the acid groups protected by the protective groups and the activated amino groups, with the amino acids in the mixture (A) found in a specific molar ratio, and a mixture (B), containing a number of y amino acids with the amino groups protected by the protective groups, with a molar ratio of the amino acids of the mixture (B) being the same as the molar ratio of the amino acids of the mixture (A), and the number x=y, and x is a figure from 11 to 18.

EFFECT: new combinations of the peptides are presented.

13 cl, 2 dwg, 1 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to a composition of a rapamycin analogue for immunomodulation and antiproliferation that involves a crystalline form of the rapamycin analogue having at least one of the following structures: and a pharmaceutically acceptable carrier. What is also described a method for preparing the crystalline form of the rapamycin analogue.

EFFECT: what is described is a new form of rapamycin, which can be used in the therapeutic treatment.

38 cl, 30 ex, 11 tbl, 21 dwg

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to field of biotechnology and immunology. Claimed is medication for treatment and prevention of TNF-dependent disorder, which contains molecule of TNF-binding nanobody, lyoprotector, surface-active substance and buffer, method of such medication obtaining, method of lyophilised preparation reduction and method of analysis of medication-manufacturing process, as well as method and set for treatment or prevention of TNF-dependent disorder.

EFFECT: invention ensures obtaining stable pharmaceutical preparations of TNF-binding nanobodies and can be applied in therapy of TNF-dependent diseases.

30 cl, 12 ex, 3 tbl, 32 dwg

FIELD: medicine, pharmaceutics.

SUBSTANCE: what is presented is the use of 7-chlor-3-(5-dimethylaminomethyl-[1,2,4]oxadiazol-3-yl)-5-methyl-4,5-dihydro-imidazo[1,5-a][1,4]benzodiazepin-6-one or a pharmaceutically acceptable salt thereof for treating various types of insomnia (terminal one, insomnia in an individual under min. 65 years, for the relief of wakefulness after the beginning of a sleep, for the prolongation of total sleep time after the beginning of a sleep - versions), pharmaceutical compositions for the appropriate application (versions) and methods of treating the various types of insomnia (versions).

EFFECT: invention is effective in treating insomnia, maintaining the sleep, terminal insomnia, including in the aged; the compound has a short half-life, ie 3-4 hours (so it causes no residual sedative action), and improves the daily activity in the aged suffering diurnal drowsiness.

61 cl, 29 dwg, 10 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to a percutaneously absorbable preparation comprising 2-[(1-benzylpiperidin-4-yl)methyl]-5,6~dimethoxyindan-1-one and/or its hydrochloride in a pressure-sensitive adhesive layer, wherein the percutaneously absorbable preparation may be administered into a patient so that Cmax per a surface area unit of the percutaneously absorbable preparation is 0.025 to 0.5 ng/ml·cm2 in the plasma concentration profile. The preparation has a favourable plasma concentration profile of 2-[(1-benzylpiperidin-4-yl)methyl]-5,6~dimethoxyindan-1-one and/or its hydrochloride with no jump of the plasma concentration.

EFFECT: what is presented is the percutaneously absorbable preparation.

29 cl, 1 tbl, 2 dwg, 4 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine and may be used to improve the adaptation possibilities and to correct the psychofunctional state in the patients with harmful working conditions. That is ensured by recommending a relaxed or partial load or motion state. Health food from Diet No.5, intake of low-mineralised (3.7 g/dm3) low-carbonate sulphate-hydrocarbonate sodium-calcium mineral water of 'Slavyanovskaya' source (Zheleznovodsk mineral water), 3.3 g/kg of body weight (200-250 ml) per one intake, 45 minutes before meals, 3 times a day. The patient takes baths with mineral water of the same composition at water temperature 36-37°C for 15 minutes, every second day, in the therapeutic course consisting of 10 procedures. The aromatherapy-assisted psychological autotraining sessions are prescribed for 14 days. Furthermore, Adaptol is prescribed for 14-day intake in a dose of 500 mg 3 times a day.

EFFECT: invention provides an improved adaptation, autonomic regulation, peroxide homeostasis, digestive functions, mental and emotional status of the patients exposed to the adverse effects of environmental factors due to the integrated treatment.

5 tbl, 3 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine and may be used to improve the adaptation possibilities and to correct the psychofunctional state in the patients with harmful working conditions. That is ensured by recommending a relaxed or partial load or motion state. Health food from Diet No.5, intake of low-mineralised (3.7 g/dm3) low-carbonate sulphate-hydrocarbonate sodium-calcium mineral water of 'Slavyanovskaya' source (Zheleznovodsk mineral water), 3.3 g/kg of body weight (200-250 ml) per one intake, 45 minutes before meals, 3 times a day. The patient takes baths with mineral water of the same composition at water temperature 36-37°C for 15 minutes, every second day, in the therapeutic course consisting of 10 procedures. The aromatherapy-assisted psychological autotraining sessions are prescribed for 14 days. Furthermore, Adaptol is prescribed for 14-day intake in a dose of 500 mg 3 times a day.

EFFECT: invention provides an improved adaptation, autonomic regulation, peroxide homeostasis, digestive functions, mental and emotional status of the patients exposed to the adverse effects of environmental factors due to the integrated treatment.

5 tbl, 3 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to solid fast-disintegrating dosage form of medication with antiparkinsonian action, which contains as active pharmaceutical ingredient memantine and/or memantine hydrochloride and cellulose II with the following ingredient ratio, wt %: memantine and/or memantine hydrochloride - 5-10, cellulose II - 90-95. Dosage form can represent pellet, created by method of direct pelletting.

EFFECT: obtaining fast-disintegrating in oral cavity dosage form, its distribution throughout oral cavity and delay of its transport to stomach, in order to provide medication delivery, not entering gastrointestinal tract and eliminating metabolism of medication in liver.

2 cl, 2 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: claimed invention relates to method of obtaining aripiprazole suspension. Method includes the following stages: (a) combining aripiprazole mass and carrier with formation of primary suspension; (b) first refining of primary suspension with obtaining secondary suspension using apparatus for refining with large shearing force, dispersant, in which shearing force is applied for material processing, or high-pressure homogeniser; and (c) second refining of secondary suspension with obtaining final sterile suspension by high-pressure homogeniser. Claimed composition also relates to methods of obtaining lyophilysed composition from aripiprazole suspension.

EFFECT: claimed invention provides possibility of obtaining aripiprazole suspension with the average size of particles from 1 to 10 mcm, without necessity to use special crystallisation technologies for obtaining initial aripiprazole mass and without application of vacuum mixing at the stage of combining aripiprazole mass and carrier.

33 cl, 13 tbl, 15 ex

FIELD: chemistry.

SUBSTANCE: present invention relates to novel quinoline compounds of formula (I) and physiologically acceptable acid addition salts and N oxides thereof, wherein R denotes a polycyclic group of formula (R) wherein * indicates the quinolinyl radical binding site; A denotes (CH2)a, where a equals 0, 1, 2 or 3; B denotes (CH2)b, where b equals 0, 1, 2 or 3; X' denotes (CH2)x where x equals 0, 1, 2 or 3; Y denotes (CH2)y where y equals 0, 1, 2 or 3; provided that a+b=1, 2, 3 or 4, x+y=1, 2, 3 or 4, and a+b+x+y=3, 4, 5, 6 or 7; Q denotes N; R1 denotes hydrogen, C1-C6-alkyl, C3-C6-cycloalkyl-C1-C4-alkyl, phenyl-C1-C4-alkyl, C1-C4-alkylcarbonyl, C1-C4-alkoxycarbonyl, phenoxycarbonyl or benzyloxycarbonyl, where phenyl rings in last two said groups are unsubstituted or carry 1, 2 or 3 substitutes selected from halogen, C1-C4-alkyl or C1-C4-halogenalkyl; R2 denotes hydrogen; R3 denotes hydrogen; p=0, 1 or 2; R4, if present, denotes C1-C4-alkyl and is bonded with X and/or Y, if p=2, two radicals R4, which are bonded with adjacent carbon atoms of X or Y, together can also denote a straight C2-C5-alkylene; q=0; n=0; m=0; X denotes S(O)2; which is located in position 3 of quinoline; Ar denotes a radical Ar1, wherein Ar1 is a phenyl, wherein the phenyl can be unsubstituted or can carry 1 substitute Rx wherein Rx denotes halogen, CN, C1-C6-alkyl, C1-C6-halogenalkyl, C1-C6-alkoxy, C1-C6-halogenalkoxy, C1-C6-alkylthio, C1-C6-halogenalkylthio, NRx1 Rx2, wherein Rx1 and Rx2 independently denote hydrogen, C1-C6-alkyl, or Rx1 and Rx2 together with a nitrogen atom form an N-bonded 5-, 6- or 7-member saturated heteromonocyclic ring or an N-boned 7-, 8-, 9- or 10-member saturated heterobicyclic ring, which are unsubstituted or carry 1, 2, 3 or 4 radicals selected from C1-C4-alkyl. The invention also relates to a pharmaceutical composition based on the compound of formula (I), a method of treatment using the compound of formula (I) and use of the compound of formula (I).

EFFECT: novel quinoline derivatives are obtained, which respond to modulation of the serotonin 5-HT6 receptor.

23 cl, 2 tbl, 44 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to new gamma-secretase inhibitors of formula I: , wherein L1,R1, R2,X,n and Ar have the values specified in the description, their pharmaceutically acceptable salts and solvates, as well as to pharmaceutical compositions based on these compounds for treating Alzheimer's disease and the use of drugs for gamma-secretase and beta-amyloid protein inhibition, and for treating neurodegenerative diseases such as Alzheimer's disease.

EFFECT: preparing the compound for treating the neurodegenerative diseases.

38 cl

FIELD: chemistry.

SUBSTANCE: invention refers to compounds of formula I: wherein a dash line represents an optional double bond; R1 means phenyl, naphthyl, pyridyl optionally substituted by one or two substitutes optionally substituted from halogen, C1-C6 alkyl, mono-halo C1-C6 alkyl, di-halo C1-C6 alkyl, CF3; R2 means H, methyl, halogen; R3 and R4 optionally mean, CF3, halo, C1-C3 alkyl wherein C1-C3 alkyl groups are optionally substituted by one or more halogen atoms; X=O; q=0; R5 means C1-C6 alkyl or its pharmaceutically acceptable salt.

EFFECT: compounds possess potassium channel modulating activity that enables using them in pharmaceutical compositions.

28 cl, 1 tbl, 2 dwg

FIELD: medicine.

SUBSTANCE: invention relates to medicine, namely to pediatric neurology, and can be used for rehabilitation of neurological disorders in children in case of neuroinfections. For this purpose, at the background of adequate complex and pathogenetic therapy, parenteral introduction of actovegin in acute period of disease additionally from the first days of disease cytoflavin is introduced intravenously by drop infusion in dose 0.6 ml/kg or 10 ml per day for 3-5 days, elcar perorally in dose 70-100 mg/kg of weight per day for 3-4 weeks. In the period of early reconvalescence pantogam is additionally introduced perorally in dose 50-70 mg/kg of weight per day for 4 weeks. In case if multifocal affection of brain substance is present, gliatilin is introduced intravenously by drop infusion in dose 1 ml per 5 kg of body weight per day in combination with intramuscular introduction of ipidacrine in dose 5-15 mg per day for 7-10 days, after that gliatilin perorally in dose 50 mg/kg of weight per day together with ipidacrine inside in dose 1 mg/kg per day for 4 weeks.

EFFECT: method makes it possible to improve disease outcome due to reduction of frequency of residual neurological deficiency formation with reduction of term of hospital treatment.

3 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, particularly, the use of human lactoferrin apo-form as an antihypoxant and a hypoxia inducible factor-1 alpha stabiliser.

EFFECT: invention provides the use of the natural iron chelator lactoferrin, no toxicity, hypoallergenicity, an ability to penetrate through the bowel into the blood flow and through the blood-brain barrier.

2 tbl, 1 dwg

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