Combined therapy using inhibitors of tyrosine kinase receptors and inhibitors of angiogenesis

FIELD: medicine, oncology, chemical-pharmaceutical industry.

SUBSTANCE: invention relates to a pharmaceutical composition designated for treatment of tumors and tumor metastasis. The composition comprises: (i) at least one antibody or its functionally intact derivative comprising a binding site that binds with the epitope ErbB1(Herl1) of receptor and (ii) at least one agent inhibiting angiogenesis being optionally in common with a pharmaceutically acceptable carrier, diluting agent or excipient used in the combined therapy in treatment of tumors and tumor metastasis. Using the proposed composition can result to the possible synergetic enhancing inhibitory effect of each specific therapeutic agent with respect to inhibition of tumor cells proliferation and providing the enhanced effective treatment as compared with individual agents using separately.

EFFECT: enhanced effectiveness of therapy.

25 cl, 1 ex

 

Technical field of invention

The invention relates to a combination therapy in the treatment of tumors and tumor metastasis, including the appointment of antagonists/inhibitors of the receptor tyrosine kinase, especially of the ErbB receptor antagonists, more preferred antagonists and anti-angiogenic means of EGF receptor (Her-1), preferably integrin antagonists, optionally together with means or forms of therapy that have additive or synergistic effect when they are used together with the specified combination of antagonists/inhibitors, such as chemotherapeutic drugs and/or radiation therapy. Proposed therapy may lead to a possible synergistic increase in effect of inhibiting the proliferation of tumor cells of each particular therapeutic agent, providing a more effective treatment than in the application of individual tools separately.

Background of the invention

The receptor for epidermal growth factor (EGF receptor or EGFR), also known as c-erbB1/Her 1, and the product of the neu oncogene (also known as C-erbB2/Her-2) are members of the superfamily, EFG receptor, which belongs to a large family of receptor tyrosine kinase. They interact on the surface of cells with specific growth factors or natural ligands, such as EGF or TGF alpha, so chivera receptor tyrosine kinase. A cascade of downstream signaling proteins are activated, which in General leads to changes in gene expression and an increase in the rate of growth.

C-erbB2 (Her-2) is a transmembrane tyrosinekinase having a molecular weight of about 185000, with significant homology to the EGF receptor (Her-1), although the specific ligand for Her-2 have yet to be clearly identified.

EGF receptor is a transmembrane glycoprotein, which has a molecular weight of 170000 and found in many types of epithelial cells. It is activated by at least three ligands, EGF, TGF-α (transforming growth factor alpha) and amphiregulin. As shown, epidermal growth factor (EGF) and transforming growth factor alpha (TGF-α), binds to the EGF receptor and leads to cell proliferation and tumor growth. These growth factors are not associated with Her-2 (Ulrich and Schlesinger, 1990, Cell 61, 203). Unlike several families of growth factors that cause the dimerization of the receptor due to their dimeric nature (e.g., PDGF), Monomeric growth factors such as EGF, contain two binding sites for receptors and, therefore, can cross-link two adjacent EGF receptor (Lemmon and others, 1997, EMBO J. 16, 281). Dimerization of the receptor is essential for internal stimulation of catalytic activity and to autophosphorylate receptor growth factor. It should be noted that cocktail recipes. the s protein tyrosine kinases (PTKs) are able to undergo as Homo-, and heterodimerization.

Clinical studies have shown that as the EGF receptor and c-erbB2 abundantly present in certain types of tumors, especially breast, ovarian, bladder, colon, kidney, head and neck cancer, squamous lung cancer (Mendelsohn, 1989, Cancer Cells 7, 359; Mendelsohn, 1990, Cancer Biology 1, 339). Therefore, these observations stimulated preclinical studies aimed at inhibiting the function of the EGF receptor human or C-erbB2, as new therapeutic approaches to the treatment of cancer (see, e.g., Baselga and others, 1996, J. Clin. Oncol. 14, 737; Fan and Mendelsohn, 1998, Curr. Opin. Oncol. 10, 67). It was reported that, for example, antibodies anti-EGF receptor, and antibodies anti-Her-2 show promising results in the treatment of human cancer. So, humanitariannet monoclonal antibody 4D5 (hMAb 4D5, HERCEPTIN®) - is already selling the drug.

It was shown that antibodies anti-EGF receptor, blocking EGF and TGF-α the receptor binding causes inhibition of proliferation of tumor cells. Taking into account these results, it produced a number of mouse and rat monoclonal antibodies against the EGF receptor, and tested their ability to inhibit tumor cell growth in vitro and in vivo (Modjtahedi and Dean, 1994, J. Oncology 4, 277). Humanitariannet monoclonal antibody 425 (hMAb 425) (US 5558864; EP 0531472) and chimeric monoclonal antibody 225 (cMAb 225) (Naramua and others, 1993, Cancer Immunol. Immunother. 37, 343-349, WO 96/40210)directed to the EGF receptor, has shown its effectiveness in clinical trials. S antibody demonstrated the ability to inhibit EGF-oposredovany the growth of tumor cells in vitro and suppress tumor formation of human in vivo in hairless mice. The antibody also exhibits synergism, mainly, with some chemotherapeutic agents (e.g. doxorubicin, adriamycin, Taxol, and cisplatin)to eliminate the human tumor in vivo in xenotransplantation mouse models. Ye and others (1999, Oncogene 18, 731) reported that ovarian cancer cells human ovarian can be successfully treated by a combination of cMAb 225 and hMAb 4D5.

Angiogenesis, also called neovascularization, is the process of vascularization of tissue, which includes the growth of new blood vessels in tissue. The process defined by infiltration of endothelial cells and smooth muscle cells. The process is believed to take place in any of three ways: (1) the vessel may grow from a pre-existing vessel; (2) de novo development of blood vessels can be the result of cells predecessor (the formation and development of blood vessels); or (3) existing small vessels can increase in diameter (Blood and others, 1990, Bioch. Biophys. Acta 1032, 89. Vascular endothelial cells are known to contain at least five RGD-hung is the most integrins, including the vitronectin receptor (αvβ3or αvβ5); collagen type I and IV receptor, laminin receptor, fibronectin/laminin/collagen receptor and fibronectin receptor (Davis and others, 1993, J. Cell. Biochem. 51, 206). The smooth muscle cell, as is known, contains at least six RGD-dependent integrins, including avβ3αvβ5.

Inhibition of cell adhesion, in vitro, using immunospecificity monoclonal antibodies for different subunits α or β integrin, involves vitronectin receptor αvβ3in cell adhesion of various cell types, including capillary endothelial cells (Davis and others, 1993, J. Cell. Biol. 51, 206).

Integrins class of cellular receptors, is known to bind extracellular matrix proteins and serve as a liaison cell-extracellular matrix and cell-cell interaction, related mainly to the case of cell adhesion. The integrin receptors belong to a family of proteins with a common structural properties of non-covalent heterodimeric glycoprotein complexes formed from α and β subunits. Vitronectin receptor, named for its original property preferred binding to vitronectin, as you know, refers to three different integrins, designated as avβ 1that αvβ3and αvβ5. αvβ1binds fibronectin and vitronectin. αvβ3connects most diverse ligands, including fibrin, fibrinogen, laminin, thrombospondin, vitronectin and the factor a background of Villebranda. αvβ5binds vitronectin. It is clear that there are different integrins with different biological functions as well as various integrins and subunits have a common biological specificity and function. An important area of recognition in ligand for many integrins is Tripeptide sequence arginine-glycine-aspartic acid (RGD). RGD is found in all of the ligands identified above for integrins vitronectin receptor. This RGD site recognition can be simulated linear and tsiklicheskimi (poly) peptides that contain the RGD sequence. Such RGD peptides, are known to be inhibitors or antagonists, respectively, the functions of the integrin. However, it is important to note that depending on the sequence and structure of the RGD peptide specificity of inhibition can be modified target specific integrins. Covered various RGD polypeptides with different integrin-specificity, for example, Cheresh, and others, 1989, Cell 58, 945, Aumailley and others, 1991, FEBS Letts. 291, 50, in numerous the applications for patents and patents (for example, patents US 4517686, 4578079, 4589881, 4614517, 4661111, 4792525; EP 0770622).

The formation of new blood vessels or angiogenesis plays a key role in the growth of malignant diseases and is of great interest in the development of tools that inhibit angiogenesis (see, for example, Holmgren and others, 1995, Nature Medicine 1, 149; Folkman, 1995, Nature Medicine 1, 27; O'reilly and others, 1994, Cell 79, 315). Use αvβ3antagonists of integrin to inhibit angiogenesis known methods of inhibiting growth of a solid tumor, reducing the blood flow to solid tumors (see, for example, US 5753230 and US 5766591 which describe the use of αvβ3antagonists, such as synthetic polypeptides, monoclonal antibodies and mimetics αvβ3that are associated with αvβ3receptor and inhibit angiogenesis). Methods and compositions for inhibition αvβ5eposredstvennoe of angiogenesis tissue, using antagonists of vitronectin receptor αvβ5described in WO 97/45447. Angiogenesis is characterized by invasion, migration and proliferation of endothelial cells, processes that depend on the interactions of cells with extracellular matrix components. In this context, the integrin cell-matrix receptors posredstwom distribution and migration of cells. As the show is but receptors of endothelial adhesion integrin αvβ3play a key role in ensuring vascular-specific targeting strategy of anti-angiogenic treatment (Brooks and others, 1994, Science 264, 569; Friedlander and others, 1995, Science 270).

The requirement for integrin αvβ3vessels in angiogenesis has been demonstrated in several models in vivo, where the formation of new blood vessels when transplantirovali human cancers has been completely suppressed or systematic introduction of peptide antagonists of integrin αvβ3and αvβ5as described above, or, alternatively, anti-αvβ3antibody LM609 (Brooks and others, 1994, Cell 79, 1157; ATS HB 9537). These antibodies block αvβ3the integrin receptor, activation of which by means of its natural ligands promotes apoptosis of proliferative angiogenic vascular cells and, thus, interrupts the formation of new blood vessels, which is a significant event for proliferatio tumors. However, as was recently reported that melanoma cells are capable of forming poutine-like structure of blood vessels, even in the absence of endothelial cells (1999, Science 285, 14), implies that the tumor is able to circumvent the effect of antiangiogenic drugs, which effect is active only in the presence of endothelial tissue.

Numerous molecules stimulate endothelial proliferation, migration and education, including VEGF, Ang1 and bFGF, and are vital factors of survival. VEGF (endothelial growth factor vascular) was identified as a selective angiogenic growth factor that can stimulate endothelial cell mitogenesis. VEGF, in particular, is a major mediator of angiogenesis in the primary tumor and ischemic ocular diseases. VEGF - glycosilated (MM: 46000), which is the endothelial kletka-specific angiogram ((Ferrara and others, 1992, Endocrin. Rev., 13, 18) and vasopressin factor (Senger and others, 1986, Cancer Res., 465629), which binds with high affinity membrane-bound receptors with tyrosinekinase activity (Jakeman and others, 1992, J. Clin. Invest., 89, 244). The tumor biopsy man show increased expression of VEGF mRNA in malignant cells and VEGF receptor mRNA in adjacent endothelial cells. VEGF expression is most evident in the areas of tumors adjacent to areas of necrosis of blood vessels (for review, see Thomas and others, 1996, J. Biol. Chem. 271(2), 603; Folkman, 1995, Nature Medicine 1, 27).

WO 97/45447 offers the use of αvβ5integrin with neovascularisation caused by VEGF, EGF and TGF-αand describes what αvβ5the antagonist can inhibit VEGF induced angiogenesis. Effectively the active anti-tumour therapy is also available when using the target VEGF receptor to inhibition of angiogenesis, using monoclonal antibodies (Witte and others, 1998, Cancer Metastasis Rev. 17(2), 155). MAb DC-101, as is known, inhibits angiogenesis of tumor cells.

In summary, it is obvious that EGF, VEGF and integrins αvβ3and αvβ5and their receptors are mainly involved in proliferation and tumor angiogenesis of the tumor, and that effective inhibitors, particularly monoclonal antibodies, directed to EGF receptor and/or VEGF receptor and/or integrin receptors or any other protein receptor tyrosine kinases are appropriate candidates for treatment of the tumor. Monoclonal antibodies specific to recognize their antigen epitopes on the relevant receptors are of particular interest.

However, the use of such antibodies, which were successful in vitro and in animal models, have not shown satisfactory performance on patients with mono-drug therapy. Similar results were obtained when other anti-angiogenetic antagonists or antagonists of EGF receptor than antibodies, have been used in clinical experiments. It seems that the tumor, while blocking some active centers can use other molecules of the cell surface to compensate for the specified initial blocking. Thus, tumors do not shrink during various anti-angiogenic or anti-proliferative therapies. For these reasons have been proposed combined therapy, in order to solve the problem using monoclonal antibodies with cytotoxic or chemotherapeutic drugs or in combination with radiation therapy. Indeed, clinical trials have demonstrated that this combined therapy is more effective than the corresponding monophosphate. So, for example, was described antibody-cytokine combined protein therapy, which causes immune-mediated reaction of oppression known tumors, such as metastatic carcinoma.

For example, the cytokine interleukin 2 (IL-2) was attached to a specific monoclonal antibody KS1/4 and ch14.18 directed to tumor-associated antigens of epithelial cell adhesion molecule (EP-ITSELF, KSA, KS1/4 antigen) or disialogangliosides GD, respectively, to form the Association of proteins ch14.18-IL-2 and KS1/4-IL-2, respectively (US 5650150). Another clinical approach based on the use of monoclonal antibodies s in combination with Herceptin®(Ye and others, 1999, I.c). Furthermore, the combination of antibodies anti-EGF receptor together with antitumor means, such as cisplatin or doxorubicin, have been described in EP 0667165 (A1) and US 6217866; such combinations, especially the combination of Herceptin®with cisplatin and other cytotoxic factors, has been described in Genentch's US 5770195. Synergistic effects between anti-angiogenic antagonist of integrin αvand the above-mentioned antibody-cytokine United proteins was observed in tumor metastasis (Lode and others, 1999, Proc. Natl. Acad. Sci. 96, 1591, WO 00/47228). Methods of use of integrin antagonists together with antitumor means recently been claimed in WO 00/38665. It was recently found that the combination of gemcitabine with a specific monoclonal antibody DC-101, that inhibits angiogenesis, increased antitumor effect in cancer of the pancreas of the mouse, compared with the use of only one of gemcitabine. DE 19842415 discloses a combination of a specific cyclic RGD peptide as integrin inhibitor, with a specific anti-angiogenic agents. Other approaches propose the use of EGF receptor blocking drugs, containing antibody or integrin antagonists in conjunction with radiation or radiation therapy, respectively (e.g. WO 99/60023, WO 00/0038715).

Although various combination therapies are at the stage of research and clinical trials, the results of these clinical trials are not fruitful. Therefore, it is necessary to develop other combinations, which can lead to increase effectiveness and reduce side effects.

Brief description of the invention

The present invention in which he describes a new pharmaceutical treatment based on a new concept for therapy of tumors for use in individual therapy a therapeutically effective amount of an agent that blocks or inhibits the receptor tyrosine kinase, preferably ErbB receptor, and more preferably EGF receptor with an anti-angiogenic agent, the composition according to the invention optionally may include other therapeutically active compounds, preferably selected from the group consisting of cytotoxic agents, chemotherapeutic agents and other pharmacologically active compounds that can enhance the efficiency of these funds or to reduce the side effects of these substances.

Thus, the invention relates to pharmaceutical compositions containing as the preferred antagonists ErbB receptor anti-EGFR (ErbB1/Her 1) antibodies and, as antiangiogenic agent inhibitor or antagonist of any one of the αvβ3that αvβ5or αvβ6integrin receptors, preferably RGD-containing linear or cyclic peptide. In particular, the invention relates, as a preferred embodiment, the specific combination therapy containing anti-EGFR or anti-Her-2 antibodies, such as humanitariannet monoclonal antibody 425 (h425, EMD 72000), chimeric mono is analnoe antibody 225 (s) or Herceptin ®together with preferably RGD-containing integrin inhibitors, preferably cyclic peptide cyclo-(Arg-Gly-Asp-DPhe-NMe-Val), optionally together with a chemotherapeutic compound.

According to this invention, these therapeutically active funds can also be supported by a pharmaceutical kit comprising a package containing one or more antagonists of the receptor tyrosine kinase, one or more anti-angiogenic agents, and optionally one or more cytotoxic/chemotherapeutic agents in a single package or in separate packages, therapy with such combinations may include, optional, and radiation therapy.

However, the invention relates, furthermore, to a combination therapy comprising the use of only one (combined) molecules with antireceptor tyrosine kinase, preferably anti-ErbB receptor activity and anti-angiogenic activity, optionally with one or more cytotoxic / chemotherapeutic agents. An example is an anti-EGFR antibody, such as h425 or s as described above and below, which is attached To the end of its Fc portion to anti-hormonal agent known recombinant or chemical methods. The following example bespecifically antibody in which one specificity directed the nuclear hormone receptor, and the other directed to the EGF receptor.

Mainly, the application may be accompanied by radiation therapy, and radiation therapy may be done simultaneously, before or after drug administration. The use of different means of combination therapy, according to the invention can also be achieved, mainly, simultaneously or sequentially. Tumors have receptors on the surfaces of their cells, which are involved in the formation of blood vessels of the tumor can be successfully treated with combination therapy of this invention.

It is known that tumors identify alternative ways for its development and growth. If one path is blocked, they often have the ability to switch to another path, seeking and using other receptors and routes of transmission signals. Therefore, the pharmaceutical combinations of the present invention can block a few of these possible strategies for the development of the tumor and, therefore, to provide a variety of useful results. The combination, according to the present invention, useful in the treatment and prevention of tumors, tumor-like growths and neoplasia disorders, tumor metastasis, which develop and grow activation of their respective hormone receptors that are present on the surface of tumor cells. predpochtitelno, various combined means of the present invention are used in combination at low dose, i.e. the dose lower than traditionally used in the clinic. The benefit of lowering the dose of the compounds, compositions, tools, and therapies of the present invention applied by the patient, comprises the reduction of adverse effects associated with higher dosages. For example, when lower doses means described above and below, there was a reduction in the frequency and severity of nausea, stop vomiting, compared with that observed at higher doses. Reducing the scope of adverse effects, there was an improvement in the quality of life of a cancer patient. Other benefits from reducing the scope of adverse effects leads to the improvement of health, reduction in the number of hospitalizations for treatment of adverse effects, and reduce the use of analgesics for the treatment of painful symptoms associated with adverse effects. Alternatively, the methods and the combination of the present invention can also maximize therapeutic effect in higher doses.

Tumor bearing (super-expressive) ErbB receptors, preferably ErbB1 (Her-1, EGFR or ErbB2 (Her-2) receptors on the surfaces of their cells, can be successfully treated by combination with the according to the invention. Combinations within the pharmaceutical treatment according to the invention show a surprising synergistic effect. When using a combination of drugs was observed real shrinkage and degradation of the tumor during clinical trials, while no significant adverse reactions to the medicinal product was not detected. First of all, a combination of three drugs (receptor tyrosine kinase, preferably the locking means ErbB receptor plus anti-angiogenic agent, plus chemotherapeutic agent) show superior performance. However, does chemotherapeutic drug synergistic effect or not, depends on the funds, the receptor tyrosinekinase, preferably antagonist of ErbB receptor and tumor cell that is subjected to the above treatment means should be checked usually from case to case.

In particular, the invention relates to:

pharmaceutical compositions containing the tool or tools that have a

(i) at least one blocking / inhibitory specificity of receptor tyrosine kinase;

(ii) at least one blocking / inhibitory specificity for angiogenesis, where the specified agent or agents is / are not cytokine immunoconjugates,optionally together with a pharmaceutically acceptable carrier, the diluent or recipient;

- as a first alternative, a pharmaceutical agent, including

(1) at least one tool having a blocking specificity to the receptor tyrosine kinase, and

(2) at least one agent with inhibitory specificity for angiogenesis;

as a second alternative, the pharmaceutical composition comprising a tool having a blocking specificity to the receptor tyrosine kinase, as well as possessing inhibitory specificity for angiogenesis;

- appropriate compositions which additionally contain at least one cytotoxic, preferably chemotherapeutic agent;

more in detail, the pharmaceutical composition in which the specified agent (i) has

blocking / inhibitory specificity to ErbB receptor;

- corresponding pharmaceutical compositions, in which the specificity of the specified funds to the ErbB receptor belongs to the EGF receptor (ErbB1/Her1) or ErbB2/Her2 receptor;

more in detail, the pharmaceutical composition in which the specified agent is an antibody or a functionally intact derivative containing the binding site that binds to an epitope ErbB1 (Her1) or Erb2 (Her2) receptor;

as a preferred embodiment, pharmaceutical compositions, inwhich the specified antibody or a functionally intact derivative selected from the group which includes:

- humanitariannet monoclonal antibody 425 (h425);

- chimeric monoclonal antibody 225 (s);

- humanitariannet monoclonal antibody Her-2, corresponding humanized, chimeric or de-immunosorbant functionally intact derivative is included;

- corresponding pharmaceutical compositions in which the above-mentioned inhibiting angiogenesis agent is αvβ3that αvβ5or αvβ6integrin inhibitor;

- corresponding pharmaceutical compositions, in which the specified integrin inhibitor is an RGD-containing linear or cyclic peptide, preferably cyclo-(Arg-Gly-Asp-DPhe-NMeVal);

as a specific embodiment, pharmaceutical compositions, in which the indicated antibody or a functionally intact derivative is a humanized monoclonal antibody 425 (h425) or chimeric monoclonal antibody 225 (s), including de-immunized form, and the above integrin inhibitor is cyclo-(Arg-Gly-Asp-DPhe-NMeVal), optionally containing, optionally in separate containers or packages, chemotherapeutic agent, which is selected from any compound group, which includes the following: cisplatin, doxorubicin, gemcitabine, docetaxel, paclitaxel, bleomycin;

- compliance is adequate pharmaceutical composition, in which the specified integrin inhibitor is an antibody or a functionally intact derivative, which includes the binding site to bind to the epitope integrin receptor, preferably selected from the group of antibodies: LM609, P1F6, E, 14D9.F8, humanized, chimeric and de-immunized version;

pharmaceutical compositions in which one of these tools is bespecifically antibody or molecule heterodontidae comprising a first binding site that binds to an epitope of the receptor tyrosine kinase, preferably ErbB receptor and a second binding site that binds to an epitope of the receptor angiogenesis, preferably integrin receptor;

specific corresponding pharmaceutical compositions in which these monoclonal antibodies are selected from h425, c225 or Her-2, and of the monoclonal antibody LM609, P1F6, E and 14D9.F8;

pharmaceutical compositions in which one of these tools is immunoconjugate comprising the antibody or antibody fragment carrying one of these blocking specificdate, and non-immunological molecules attached to the antibody or antibody fragment bearing a different specificity;

- corresponding pharmaceutical compositions, in which part of the antibody or its fragment contains the binding site, which binds to the epitope ErbB receptor, preferably EGF receptor (Her-1), and connected non-immunological molecule contains a binding site that binds to an epitope of an integrin receptor;

its specific pharmaceutical compositions in which the specified portion of the antibody that binds to the epitope ErbB receptor, selected from monoclonal antibodies h425, c225 or Her-2, and the specified non-immunological part that binds to the epitope of an integrin receptor, is cyclo-(Arg-Gly-Asp-DPhe-NMeVal);

- pharmaceutical collection, which includes

(i) a package containing at least one receptor, inhibiting tyrosinekinase, preferably by means of a blocking ErbB receptor, and

(ii) a package containing at least one tool, inhibiting angiogenesis, preferably the tool, inhibitory αvβ3that αvβ5or αvβ6the integrin receptor, more preferably RGD-containing linear or cyclic peptide, in particular cyclo-(Arg-Gly-Asp-DPhe-NMeVal);

optional comprising packaging containing cytotoxic agent;

- corresponding pharmaceutical kit, in which the tool blocking ErbB receptor is an antibody or a functionally intact derivative having a binding site which binds to an epitope of a specified receptor; the specified antibody is preferably selected from the group of antibodies, which includes:

humanitariannet monoclonal antibody 425 (h425), a chimeric monoclonal antibody 225 (s) or humanitariannet monoclonal antibody to Her-2;

- pharmaceutical set in which the specified tool, inhibiting angiogenesis, is an antibody or its active derivative, which is preferably chosen from the group of antibodies comprising: LM609, RN, E and 14D9.F8;

a specific embodiment of the invention, the specific pharmaceutical kit containing

(i) a package containing humanitariannet monoclonal antibody 425 (h425), a chimeric monoclonal antibody 225 (s), or a functionally intact derivative, and

(ii) a package containing cyclo-(Arg-Gly-Asp-DPhe-NMeVal), optionally containing chemotherapeutic agent selected from any connection group, which includes the following: cisplatin, doxorubicin, gemcitabine, docetaxel, paclitaxel, bleomycin;

- the use of a pharmaceutical composition or a pharmaceutical kit as above, below and in the claims, for the manufacture of a medicinal product for the treatment of tumors and tumor metastases;

- pharmaceutical treatment or a method of treating tumors or tumor metastases of a patient, which comprises applying to the patient therapeutics and an effective amount of vehicle or vehicles, with

(i) at least one blocking the specificity of receptor tyrosine kinase, and

(ii) at least one inhibitory specificity for angiogenesis,

where the specified agent or agents is/are not cytokine immunoconjugates, optionally, together with cytotoxic, preferably a chemotherapeutic agent, and where, preferably, the said means (i) an antibody or a functionally intact derivative containing the binding site that binds to an epitope ErbB receptor, preferably ErbB1 (Her1) or rb2 (Her2) receptor, and the specified agent (ii) is αvβ3that αvβ5or αvβ6integrin inhibitor or vehicle, blocking VEGF receptor; and, finally,

- appropriate way, in which the indicated antibody directed at the ErbB receptor is chosen from the group which includes: humanitariannet monoclonal antibody 425 (h425), a chimeric monoclonal antibody 225 (s) or humanitariannet monoclonal antibody Her-2, and anti-angiogenic agent is a cyclo-(Arg-Gly-Asp-DPhe-NMeVal), optionally together with a cytotoxic drug selected from the group: cisplatin, doxorubicin, gemcitabine, docetaxel, paclitaxel, bleomycin.

Pharmaceutical cured the e using pharmaceutical compositions and kits according to the invention, may be accompanied, simultaneously or sequentially with radiation therapy.

Mainly, according to the invention, it is possible to note the four different combinations of pharmaceutical compositions:

(i) means containing at least one blocking activity/specificity to the receptor tyrosine kinase, preferably ErbB receptor, combined with means containing at least one anti-angiogenic activity (combination of two drugs);

(ii) means containing at least one blocking activity/specificity to the receptor tyrosine kinase, preferably ErbB receptor, combined with means containing at least one anti-angiogenic activity, combined with at least one chemotherapeutic agent (a combination of three drugs);

(iii) means containing at least one blocking activity/specificity to the receptor tyrosine kinase, preferably ErbB receptor, and at least one anti-angiogenic activity, combined in one molecule (the combination of one medicinal product with the activity of two drugs);

(iv) means containing at least one blocking activity/special which was mentioned to the receptor tyrosine kinase, preferably ErbB receptor, and at least one anti-angiogenic activity, combined in one molecule, combined with at least one chemotherapeutic agent (a combination of two drugs that have activity three drugs).

Funds may be applied simultaneously or sequentially in any of these cases. According to the above, the methods of the invention include primarily the following combinations:

(i) means containing at least one blocking activity/specificity to the receptor tyrosine kinase, preferably ErbB receptor, combined with means containing at least one anti-angiogenic activity (appointment of two drugs);

(ii) means containing at least one blocking activity/specificity to the receptor tyrosine kinase, preferably ErbB receptor, combined with means containing at least one anti-angiogenic activity (appointment of two drugs)and radiation therapy;

(iii) means containing at least one blocking activity/specificity to the receptor tyrosine kinase, preferably ErbB receptor, combined with means containing at least one anti-angiogenic activity, combined with at least about what it chemotherapeutic agent (the appointment of three drugs);

(iv) means containing at least one blocking activity/specificity to the receptor tyrosine kinase, preferably ErbB receptor, combined with means containing at least one anti-angiogenic activity, combined with at least one chemotherapeutic agent (the appointment of three drugs)and radiation therapy;

(v) means containing at least one blocking activity/specificity to the receptor tyrosine kinase, preferably ErbB receptor, and at least one anti-angiogenic activity, combined in one molecule (the assignment of one medicinal product with the activity of two drugs);

(vi) means containing at least one blocking activity/specificity to the receptor tyrosine kinase, preferably ErbB receptor, and at least one anti-angiogenic activity, combined in one molecule (the assignment of one medicinal product with the activity of two drugs)and radiation therapy;

(vii) means containing at least one blocking activity/specificity to the receptor tyrosine kinase, preferably ErbB receptor, and at least one anti-angiogenic activity, combined in one molecule, combined with at least one hee is iterations tool (the appointment of two medicines, having the activity of three drugs);

(viii) means containing at least one blocking activity/specificity to the receptor tyrosine kinase, preferably ErbB receptor, and at least one anti-angiogenic activity, combined in one molecule, combined with at least one chemotherapeutic agent (the appointment of two drugs that have activity three drugs)and radiation therapy;

Pharmaceutical combinations and methods of the present invention provide various advantages. The combination, according to the present invention, useful in the treatment and prevention of tumors, tumor-like diseases and neoplasia violations. Preferably, various combined means of the present invention used in combination in low doses, i.e. doses lower than the conventionally used in clinical situations. The advantage of reducing the dose of the compounds, compositions, tools, and therapies of the present invention, assigned to a mammal, includes the reduction of adverse factors associated with higher doses. For example, the reduction in the dosage of a chemotherapeutic drug, such as methotrexate, doxorubicin, gemcitabine, docetaxel, paclitaxel, bleomycin or cisplatin leads to reduction is astate and severity of nausea and vomiting, compared with the observed at higher dosages. In the present invention are considered similar advantages to compounds, compositions, tools, and therapies in combination with antagonists of the integrin. Lowering the sphere of influence of adverse factors, there was an improvement in the quality of life of a cancer patient. Other benefits from reducing the scope of adverse factors lead to the improvement of health, reduction in the number of hospitalizations for treatment of adverse factors, and reduce the use of analgesics for treatment of pain associated with adverse factors.

Alternatively, the methods and the combination of the present invention can also maximize therapeutic effect in higher doses.

A detailed description of the invention

Unless otherwise specified, the selected terms and phrases used in this invention have the meanings and definitions as below. In addition, these definitions and values describe the invention in more detail, including the preferred options of the description.

"Receptor" or "receptor molecule" is soluble or associated/associated membrane protein or glycoprotein, consisting of one or more domains, which binds the ligand, to form the idea is with the receptor-ligand. Linking the ligand, which may be an agonist or antagonist, a receptor is activated or inactivated, and can activate or block the signal.

The term "ligand" or "receptor ligand" understand natural or synthetic compound that binds to a receptor molecule to form a complex receptor-ligand. The term ligand includes the concept of agonists, antagonists and compounds with partial agonist/antagonist action.

"Agonist" or "receptor agonist" is a natural or synthetic compound that binds to a receptor to form a receptor-agonist complex, activating the said receptor and the complex receptor agonist, respectively, initiating the occurrence of the signal and the emergence of biological processes.

The term "antagonist" or "receptor antagonist" understand natural or synthetic compound that has a biological effect opposite to the effect of the agonist. The antagonist binds to a receptor and blocks the action of receptor agonist, competing with the agonist for the receptor. The antagonist is determined by its ability to block the actions of the agonist. Receptor antagonist may be an antibody or immunotherapy effective fragment. Preferred antagonists according to this invention, p is precisley and analyzed below.

"ErbB receptor" is a protein tyrosine kinase receptor, which belongs to the group of ErbB receptors and includes EGFR (ErbB1), ErbB2, ErbB3 and ErbB4 receptors and other members of this group, which will be identified in the future. ErbB receptor mainly consists of the extracellular domain, which may bind ErbB ligand; lipophilic transmembrane domain; saved intracellular tyrosine kinase domain; and transmitting the signal domain with carboxyl end that contains multiple tyrosine residues that can be phosphorylated. ErbB receptor may be a "natural sequence" ErbB receptor, or its variant with amino acid sequence". Preferably ErbB this receptor with the natural sequence of ErbB receptor of human rights. ErbB1 is the product of a gene encoding EGFR protein. Most preferred is the EGF receptor (Her-1). The expression "ErbB1" and "Her 1" are used interchangeably and refer to a protein of human Her-1. The expression "ErbB2" and " Her-2" are used interchangeably and refer to a protein of human Her-2. Receptors ErbB1 (EGFR) are preferred according to this invention.

"ErbB ligand" is a polypeptide that binds and/or activates ErbB receptor. ErbB ligands that bind to EGFR include EGF, TGF-α, amphiregulin, betacellulin, HB-EGF and epiregulin.

The term "antagonist/inhibitor tyrosi the kinase" refers to natural or synthetic means, which are able to inhibit or block tyrosine kinase, including receptor tyrosinekinase, which are a matter of special interest to the present invention. Thus, the term includes "antagonists/inhibitors of ErbB receptor", defined below in more detail. In addition to these antagonists, preferably antibodies anti-ErbB receptor, optionally with suitable tyrosine kinase antagonists of the invention are chemical compounds that have shown efficacy in mono-drug therapy, for example, in breast cancer and prostate cancer. The corresponding tyrosine kinase inhibitors indolocarbazoles type can be obtained on the basis of information contained in documents, such as patents US 5516771; 5654427; 5461146; 5650407. Patents US 5475110; 5591855; 5594009 and WO 96/11933 describe inhibitors pyrrolopyrazole type tyrosine kinase gene and prostate cancer. Preferably, the dosage of chemical inhibitors of tyrosine kinase, as described above, lies in the range of 1 PCG/kg to 1 g/kg body weight, per day. More preferably, the dose of a tyrosine kinase inhibitor is in the range from 0.01 mg/kg to 100 mg/kg of body weight, per day.

The term "antagonist/inhibitor of ErbB receptor" refers to a natural or synthetic molecule, which binds and blocks or inhibits ErbB receptor and, therefore, is part of a group "(receptor is) antagonists/inhibitors of tyrosine kinase". Thus, blocking receptor antagonist prevents binding ErbB ligand (agonist) and activation of the complex agonist/receptor ligand. The ErbB antagonists can be directed to Her 1 (or EGFR / Her-1 or Her-2. Preferred antagonists according to the invention, directed to EGF receptor (EGFR, Her-1). Antagonist of ErbB receptor may be an antibody or immunotherapy effective fragment or fragment neimmunizirovannah molecules, such as peptide, polypeptide protein. Chemical molecules also include, however, antibodies anti-EGFR and antibodies anti-Her-2 are preferred antagonists, according to the invention.

Preferred according to the invention are antibodies anti-Her1 and anti-Her2, more preferably antibodies anti-Her1. Preferred antibodies anti-Her1 this MAb 425, preferably humanized MAb 425 (hMAb 425, US 5558864; EP 0531472) and chimeric MAb 225 (cMAb 225, US 4943533 and EP 0359282). Most preferred is a monoclonal antibody h425, which showed in the mono-drug therapy high efficiency, along with reduced adverse effects, and side effects. The most preferred antibody anti-Her2 Herceptin is®produced by Genentech/Roche.

Effective antagonists of EGF receptor according to the invention can be also other natural or si is synthetic chemical compounds. Some examples of preferred molecules of this class include organic compounds, ORGANOMETALLIC compounds, salts of organic and organometallics connections.

Effective antagonists ErbB receptor according to the invention can also be small molecules. Small molecules according to the invention is not biological molecules, as defined above, with a molecular weight of approximately not more than 400. Preferably, they do not have the protein or peptide structure, and most preferably, it is a synthetically derived chemical compounds. Some preferred examples of small molecules include organic compounds, ORGANOMETALLIC compounds, salts of organic and organometallics connections.

Numerous small molecules have been described, as are useful for the inhibition of the EGF receptor and/or Her-2 receptor. Examples are: styryl substituted heteroaryl compounds (US 5656655); bis-, mono - and/or bicykliczny the aryl heteroaryl, carbocyclic and heterocyclic connection (US 5646153); tricycline connection pyrimidine (US 5679683); derivatives of hintline having the activity of inhibiting receptor tyrosine kinase (US 5616582); derivatives heteroallergy or heteroarylboronic (US 5196446); the connection, designated as -(2,6-dichlorophenyl)-2-(4-(2-diethyl-aminoethoxy)phenylamino)-8-methyl-8H-pyrido(2,3)-5-pyrimidine-7-he (Panek, and others, 1997, J. Pharmacol. Exp. Therap. 283,1433), which inhibits EGFR, DERIVED and FGFR group of receptors.

"Anti-angiogenic agent" refers to a natural or synthetic compound that blocks or interferes to some extent, the development of blood vessels. Anti-angiogenic molecule may, for example, be a biological molecule that binds to and blocks the angiogenic growth factor or growth factor receptor. Preferred anti-angiogenic molecule when it binds to the receptor, preferably with the integrin receptor or VEGF receptor. The term includes, according to the invention, the prodrug of the specified angiogenic funds. There are many molecules with different structure and origin that exhibit antiangiogenic properties. The most important classes of angiogenic inhibitors or blocking means which are suitable in this invention are, for example:

(i) anti-Mitty, such as fluorouracil, mitomycin-C, Taxol;

(ii) the metabolites of estrogen, such as 2-methoxyestradiol;

(iii) inhibitors of matrix metalloproteinases (MMP), which inhibit zinc metalloproteinases (metalloprotease) (such as, for example, batimastat, W, TIMPs, minocycline, GM6001, or described in the "Inhibition of Matrix Metalloproteinases: Therapeutic Applications," Golub, Annals of the New York Academy of Science, Vol. 878a; Greenwald, Zucker (Eds.), 199);

(iv) anti-angiogenic multifunctional tools and factors such as IFNα (US 4,530,901; US 4,503,035; 5,231,176); angiostatin fragments of plasminogen (for example, Kringle (kringle) 1-4, Kringle 5, Kringle 1-3 (O'reilly, M.S., and others, Cell (Cambridge, Mass.) 79(2): 315-328, 1994; Cao and others, J. Biol. Chem. 271: 29461-29467, 1996; Cao and others, J. Biol Chem 272: 22924-22928, 1997); endostatin (O'reilly, M.S., and others, Cell 88(2), 277, 1997 and WO 97/15666), thrombospondin (TSP-1; Frazier, 1991, Curr Opin Cell Biol 3(5):792); platelet factor 4 (PF4);

(v) inhibitors of the plasminogen activator/urokinase;

(vi) antagonists of the receptor for urokinase;

(vii) heparinase;

(viii) analogues fumagillin, such as TNP-470;

(ix) the tyrosine kinase inhibitors, such as SUI 01 (many of above and below antagonist of ErbB receptor antagonists, EGFR / Her-2), and tyrosine kinase inhibitors, and can therefore show that anti-EGF receptor blocking activity, which is manifested in the inhibition of tumor growth, as well as anti-angiogenic activity, which is manifested in the inhibition of the development of blood vessels and endothelial cells, respectively);

(x) suramin and analogues.;

(xi) angiostatic steroids;

(xii) antagonists of VEGF and bFGF;

(xiii) antagonists, VEGF receptor, such as antibodies anti-VEGF receptor (DC-101);

(xiv) antagonists flk-1 and flt-1;

(xv) inhibitors of the cyclooxygenase-II, such as COX-II;

(xvi) integrin antagonists and antagonists of the integrin receptor, t is the cue as antagonists α v and antagonists αv receptor, for example, antibodies anti-αv receptor and RGD peptides. The integrin antagonists (receptor) are preferred, according to this invention.

The term "antagonists/inhibitors of integrin" or "antagonists/inhibitors of integrin receptor" refers to a natural or synthetic molecule that blocks and inhibits the integrin receptor. In some cases, the term includes antagonists directed to ligands of these receptors integrin (such as for αvβ3: vitronectin, fibrin, fibrinogen, factor a background of Villebranda, thrombospondin, laminin; αvβ5: vitronectin; αvβ1: fibronectin and vitronectin; αvβ6: fibronectin). Antagonists directed to integrin receptors, are preferred according to the invention. The integrin antagonists (receptor) can be natural or synthetic peptides, dipeptide, peptidomimetics, antibodies, such as antibodies or their functional fragments, or immunoconjugate (related proteins). Preferred inhibitors of integrin invention is directed to a receptor αvintegrins (for example, αvβ3that αvβ5that αvβ6and sub-classes). Predpochtitel the diversified integrin inhibitors are antagonists α vand, in particular, antagonists αvβ3. Preferred antagonists αvaccording to the invention are RGD peptides antagonists peptidomimetics (dipeptides) and antibody anti-integrin receptor, such as antibodies, blocking αvthe receptors.

Examples of non-immunological antagonists αvβ3are described in patents US 5,753,230 and US 5,766,591. Preferred antagonists include linear and cyclic RGD-containing peptides. Cyclic peptides are generally more stable and have an extended serum half-life. The most preferred integrin antagonist according to the invention is, however, cyclo-(Arg-Gly-Asp-DPhe-NMeVal) (EMD 121974, Cilengitide®, Merck KgaA, Germany; EP 0770622), which is effective in blocking the receptor integrin αvβ3that αvβ1that αvβ6that αvβ8that αvβ3. Suitable peptidyl, as well as antagonists peptidomimetic (dipeptide) receptor integrin αvβ3vβ5vβ6have been described in the scientific literature and in patents. For example, you can refer to Hoekstra and Poulter, 1998, Curr. Med. Chem. 5, 195; WO 95/32710; WO 95/37655; WO 97/01540; WO 97/37655; WO 97/45137; WO 97/41844; WO 98/08840; WO 98/18460; WO 9/18461; WO 98/25892; WO 98/31359; WO 98/30542; WO 99/15506; WO 99/15507; WO 99/31061; WO 00/06169; EP 0853084; EP 0854140; EP 0854145; US 5,780,426 and US 6,048,861. Patents that describe benzazepin, as well as related benzodiazepine and benzocycloheptene antagonists αvβ3receptor integrin, which are also suitable for use in this invention, including WO 96/00574, WO 96/00730, WO 96/06087, WO 96/26190, WO 97/24119, WO 97/24122, WO 97/24124, WO 98/15278, WO 99/05107, WO 99/06049, WO 99/15170, WO 99/15178, WO 97/34865, WO 97/01540, WO 98/30542, WO 99/11626 and WO 99/15508. Other antagonists of the integrin receptor, form the basis of the limitations of the ring structure, have been described in WO 98/08840; WO 99/30709; WO 99/30713; WO 99/31099; WO 00/09503; US 5,919,792; US 5,925,655; US 5,981,546 and US 6,017,926. In the US 6,048,861 and in WO 00/72801 describes several derivatives Romanovyh acids, which are potent antagonists αvβ3the integrin receptor. Other chemical small molecule antagonists of integrin (mainly antagonists of vitronectin) are described in WO 00/38665. Other antagonists αvβ3receptor, as shown, has shown efficacy in the inhibition of angiogenesis. For example, a synthetic receptor antagonists, such as (S)-10,11-dihydro-3-[3-(pyridine-2-ylamino)-1-propyloxy]-5H-dibenzo[a,d]Cycloheptane-10-acetic acid (known as SB-265123), were tested in several model systems mammals (Keenan and others, 1998, Bioorg. Med. Chem. Lett. 8(22), 3171; Ward and others, 1999, Drug Metab. Dispos. 27(11), 1232).

IU is Tiki analysis to identify antagonists of the integrin, used as antagonists, as described, for example, Smith and others, 1990, J. Biol. Chem. 265, 12267, and in the referenced patent literature referenced. Antibody anti-integrin receptor is also well known. Suitable monoclonal antibody anti-integrin (e.g., αvβ3that αvβ5that αvβ6) can be modified to create antigen-binding fragments, including F(ab)2, Fab and create a Fvor antibodies with one chain. One suitable and preferred monoclonal antibody directed against the receptor integrin αvβ3identified as LM609 (Brooks and others, 1994, Cell 79, 1157; ATS HB 9537). Effective specific antibody anti-αvβ5, P1F6, described in WO 97/45447, which is also preferred, according to this invention. Other suitable selective antibody αvβ6is MAb 14D9.F8 (WO 99/37683, DSM ACC2331, Merck KGaA, Germany), as well as MAb E (EP 0719859; DSM ASS, Merck KGaA), which is selectively directed to αv-chain of integrin receptors. Other acceptable antibody anti-integrin is a commercial drug Vitraxin®.

The term "antibody" or "immunoglobulin" is used here in its broadest sense and, in particular, covers intact monoclonal antibodies, polik the national antibodies multispecific antibodies (for example, bespecifically antibodies) formed from at least two intact antibodies, and fragments of antibodies, because they exhibit the desired biological activity. The term mainly includes the concept of heterogenite, which consist of two or more antibodies, or their fragments, with different binding specificity, which are connected together. Depending on the amino acid sequence of the invariant parts, intact antibodies can be correlated to various antibody (immunoglobulin) classes". There are five major classes of intact antibodies: IgA, IgD, IgE, IgG and IgM, and several of them can be further divided into "subclasses (isotypes), e.g., IgG1, IgG2, IgG3, IgG4, IgA, and IgA2. Invariant domains, consisting of heavy chains, corresponding to different classes of antibodies are called α, δ, ε, γ and μ respectively. The preferred base class for antibodies according to the invention, it is IgG, and more IgG1 and IgG2. Antibodies is mainly glycoprotein, with a molecular weight of approximately 150,000, consisting of two identical light (L) chains and two identical heavy (H) chains. Each light chain is linked to a heavy chain by one covalent disulfide bond, while the number of disulfide linkages varies between the heavy chains of RA is personal isotypes of immunoglobulin.

Each heavy and light chain also has regularly spaced megamachine disulfide bridges. Each heavy chain at one end has a variable domain (VH), complemented by near constant domains. Variables region include gipervalentnye area or "CDR" areas that contain the antigen binding site and are responsible for the specificity of the antibodies, and "FR" region, which are important in relation to the affinity/avidity antibodies. HyperTerminal region usually includes amino acid residues from a complementary defined region" or "CDR" (e.g. residues 24-34 (L1), 50-56 (L2) and 89-97 (L3) in the light chain variable domain and 31-35 (H1), 50-65 (H2) and 95-102 (H3) in the heavy chain variable domain; and/or from the remnants of the "hyperpersonal loop" (e.g. residues 26-32 (L1), 50-52 (L2) and 91-96 (L3) in the light chain variable domain and 26-32 (H1), 53-55 (H2) and 96-101 (H3) in the heavy chain variable domain; Chothia and Lesk J. Mol. Biol. 196:901-917 (1987)). "FR" residues (area of operations) are those residues of the variable domains, which differ from the remnants HyperTerminal area, as defined here. Each light chain has a variable domain at one end (VL) and a constant domain at the other end. Constant domain of the light chain is aligned with the first constant domain of the heavy chain and the variable domain of the light chain is aligned with the variable domain thee who Eloy chain. Certain amino acid residues are believed to form a bond between the variable domains of the light chain and heavy chain. The "light chains" of antibodies from any specimens of vertebrate animal can be assigned to one of two clearly distinct types, called Kappa (κ) and lambda (λ)based on amino acid sequences of their constant domains.

The term "monoclonal antibody", as used in this invention relates to an antibody obtained from a population essentially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical except possibly occurring mutations that may be present in minor amounts. Monoclonal antibodies are highly specific, targeted operate against a single antigenic site. In addition, unlike drugs polyclonal antibodies, which include different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant on the antigen. In addition to their specificity, monoclonal antibodies are preferable in that they can be synthesized uncontaminated by other antibodies. Methods for creating monoclonal antibodies include the method hybridomas described by Kohler and Milstein (1975, Nature 256,495) and in "Monoclonal Antibody Technology, The Production and Characterization of Rodent and Human Hybridomas" (1985, Burdon and others, Eds, Laboratory Techniques in Biochemistry and Molecular Biology, Volume 13, Elsevier Science Publishers, Amsterdam), or can be obtained by known recombinant DNA methods (see, for example, US 4,816,567).

Monoclonal antibodies can also be obtained from the collection of antibody phage, using the methods described, for example, Clackson and others, Nature, 352:624-628 (1991) and Marks etc., J. Mol. Biol., 222:58, 1-597(1991).

The term "chimeric antibody" means an antibody in which a portion of the heavy and/or light chain is identical with or homologous with respect to corresponding sequences in antibodies derived from a particular species or belonging to a specific class or subclass of antibody, whereas the remaining member of the chain(EC) is identical with or homologous to corresponding sequences in antibodies derived from another species or belonging to another class or subclass antibodies as well as fragments of these antibodies, as they are expected biological activity (e.g., US 4,816,567; Morrison and others, Proc. Nat. Acad. Sci. USA, 81:6851-6855 (1984)). Methods to generate chimeric and humanized antibodies are also known. For example, methods to generate chimeric antibodies include description in patents Boss (Celltech) and Cabilly (Genentech) (US 4,816,397; US 4,816,567).

"Humanized antibodies" are forms of chimeric antibodies derived from human, for example, rodent)that contain minimal sequence derived from non-human immunoglobulin. In the most part, humanized antibody is a human immunoglobulins (recipient antibody)in which residues from HyperTerminal region (CDRs) of the recipient are replaced remnants of hyperpersonal the field of non-human species (donor antibody)such as mouse, rat, rabbit, or Primate having the desired specificity, affinity and capacity. In some cases, the working region (FR) residues of the human immunoglobulin are replaced by corresponding inhuman remnants. Furthermore, humanized antibodies may contain residues that are not found in the recipient antibody or in the donor antibody. These changes were made to improve the quality of the antibodies. In General, humanitariannet antibody includes at least one, and typically two, changing the domain in which all or almost all gipervalentnye loops correspond to those of non-human immunoglobulin and all or almost all of the FRs, which correspond to the sequence of human immunoglobulin. Humanitariannet antibody optionally also will comprise at least a portion of an immunoglobulin constant region (Fc), typically from a human immunoglobulin. Methods to create the humanized antibodies is described, for example, Winter (US 5,225,539) and Boss (Celltech, US 4,816,397).

"Fragments of antibodies include a portion of an intact antibody, preferably including antigen-linked or variable region. Examples of fragments of antibodies are Fab, Fab', F(ab')2, Fv and Fc fragments, diately, linear antibodies, molecules, single chain antibodies; and multispecific antibodies formed from a fragment(s) of the antibody. "Intact" antibody is one that contains the antigen-linked variable region and the constant domain of the light chain (CL) and constant domain of the heavy chain, CN, CH2 and CH3. Preferably, the intact antibody has one or more effector functions. The papain digestion of antibodies produces two identical antigen-binding fragment, called "Fab" fragments, each comprising a single antigen-linked plot and CL and SN region, and a residual "Fc" fragment, whose name reflects its ability to crystallize. "Fc region of the antibody includes, as a rule, CH2, CH3, and the area of the loop IgG1 or IgG2 antibodies of the main class. The area of the loop group of about 15 amino acid residues, which combine IN area with the CH2-CH3 region. Treatment with pepsin gives a "F(ab')2 fragment that has two antigen-related site and still capable of antigen cross-linking. "Fv" is the minimum antibody fragment that contains the floor the first antigen-uznavaemi and the antigen-binding site. This region consists of a dimer of one heavy chain and one light chain variable domain with a tight, non-covalent bond. In this configuration, these three gipervalentnye region (CDRs) of each variable domain interact to define an antigen-binding site on the surface of the VH-VL dimer. All together, these six hyperparenting areas give antigennegative the specificity of the antibody. However, even a single variable domain (or half of an Fv, containing only three gipervalentnye areas identified for an antigen) has the ability to recognize and bind antigen, although at a lower affinity than the entire binding site. Fab fragment also contains the constant domain of the light chain and the first constant domain (SN) the heavy chain. "Fab'fragments differ from Fab fragments several additional residues carboxyl end of the heavy chain CN domain, including one or more cysteines from the area of the loop antibodies. F(ab')2 fragments of antibodies originally were produced as pairs of Fab' fragments which have a cysteine loop between them. Other chemical compounds fragments of antibodies are also known (see, for example, Hermanson, Bioconjugate Techniques, Academic Press 1996; US 4342566).

"Single chain Fv "or" scFv fragments of antibodies includes V and V domains of antibodies, where these domains are present in a Single polypeptide the th circuit. Preferably, the Fv polypeptide further comprises a polypeptide linker between the VH and VL domains, which allow scFv to form the desired structure for antigen binding. Single-chain FV antibodies are known, for example, Pluckthun (Pharmacology of Monoclonal Antibodies, Vol.113, Rosenburg and Moore eds., Springer-Verlag, New York, pp.269-315 (1994)), WO 93/16185; US 5,571,894; US 5,587,458; Huston and others (1988, Proc.Natl. Acad. Sci. 85, 5879) or Skerra and Plueckthun (1988, Science 240, 1038).

"Bespecifically antibody" is a single, divalent antibodies (or immunotherapy effective fragments)that have two different specific antigen-binding site. For example, the first antigen-binding site is aimed at angiogenesis receptor (e.g., integrin or VEGF receptor), whereas the second antigen-binding site aims to ErbB receptor (e.g., EGFR or Her-2). Bespecifically antibodies can be obtained by chemical methods (see, for example, Kranz and others (1981) Proc. Natl. Acad. Sci. USA 78, 5807), method polydoma" (see US 4,474,893), or by recombinant DNA technology, all of which are known per se. Other methods described in WO 91/00360, WO 92/05793 and WO 96/04305. Bespecifically antibodies can also be derived from a single chain antibodies (see, e.g., Huston and others (1988) Proc. Natl. Acad. Sci 85, 5879; Skerra and Plueckthun (1988) Science 240, 1038). They are analogues of variables plots antibodies, obtained as a single polypeptide chain. To get biosp nificence connecting means, single chain antibodies can be joined together chemically or by genetic engineering methods known from the prior art. You can also get bespecifically antibodies according to this invention, using leucine of zipper sequence. Used sequence derived from leucine to zipper regions of the transcription factors Fos and Jun (Landschulz and others, 1988, Science 240, 1759; for a review, see Maniatis and Abel, 1989, Nature 341, 24). Sipera leucine is certain amino acid sequence of about 20-40 residues with leucine, usually found in every seventh residue. Such sipera sequence form an amphipatic α-helix, leucine residues aligned on the hydrophobic side for the formation of the dimer. Peptides corresponding to Sitaram leucine Fos and Jun proteins form heterodimeric selectively (O'shea and others, 1989, Science 245, 646). Super-containing bespecifically antibodies and methods for obtaining them are also disclosed in WO 92/10209 and WO 93/11162. Bespecifically antibody according to the invention can be an antibody directed to VEGF receptor and αvβ3receptor, as discussed above relative to antibodies with a single specificity.

"Getrootframe are two or more antibodies or antibody-binding fragments, which are connected together, each of them has different links special is vechnosti. Heterodontidae can be obtained by conjugation of two or more antibodies or fragments of antibodies. Preferred heterodontidae consist of cross stitched Fab/Fab' fragments. The number of connecting or krasivaya agents can be used for conjugation of antibodies. Examples include protein a, carbimide, N-Succinimidyl-S-acetyl-thioacetate (SATA) and N-Succinimidyl-3-(2-pyridyldithio)propionate (SPDP) (see, e.g., Karpovsky and others (1984) J. EXP. Med. 160, 1686; Liu et a. (1985) Proc. Natl. Acad. Sci. USA 82, 8648). Other methods include described Paulus, Behring Inst. Mitt., No. 78, 118 (1985); Brennan and others (1985) Science 30 m:81 or Glennie and others (1987) J. Immunol. 139, 2367. Another method uses on-phenylenedimaleimide (oPDM) to link the three Fab' fragments (WO 91/03493). In the context of this invention, multispecific antibodies are also suitable and can be obtained, for example, according to WO 94/13804 and WO 98/50431.

The term "fusion protein" refers to a natural or synthetic molecule, consisting of one or more proteins or peptides or their fragments having different specificity, which are connected together arbitrarily linker molecule. A specific embodiment of the term includes the construction of a merger where at least one protein or peptide is an immunoglobulin or antibody, respectively, or any part thereof ("immunoconjugate").

The term "immunoconjugate" refers to the antibody or immuno is Abolina, respectively, or their immunologically effective fragment that is attached covalent bond to non-immunological active molecule. Preferably, the binding partner is a peptide or protein that can be glycosylated. This molecule is not an antibody may be linked to the C-terminal constant heavy chain antibodies or with an N-terminal variable light and/or heavy chains. Liaudies parts can be connected by linker molecules, which, as a rule, is a peptide containing from 3 to 15 amino acid residues. Immunoconjugate, according to the invention, consist of immunoglobulin or immunotherapy active fragment directed to a receptor tyrosine kinase, preferably ErbB (ErbB1/ErbB2) receptor and integrin antagonistic peptide, or angiogenic receptor, preferably integrin or VEGF receptor and TNFα or the fusion protein, consisting essentially of TNFα and IFNγ or another. a suitable cytokine, which is connected to its N-terminal to C-terminal specified immunoglobulin, preferably the Fc portion. The term also includes the corresponding merge structures containing bi - or multispecific immunoglobulins (antibodies) or fragments thereof.

The term "functionally intact derivative" means, according to the invention, a fragment or frequent the, modified, variant, homologue or de-immunized form (modification, where the epitopes that are responsible for immune responses removed) compound, peptide, protein, antibody (immunoglobulin), immunoconjugate etc. that has mostly the same biological and/or therapeutic function compared to the original compound, peptide, protein, antibody (immunoglobulin), immunoconjugates, etc. However, the term also includes such derivative compounds that identifies a reduced or increased efficiency.

The term "cytokine" is a generic term for proteins released by the cell population that interact with another cell as intercellular mediators. Examples of such cytokines are lymphokines, Monokini and traditional polypeptide hormones. Included among the cytokines, growth hormones such as human growth hormone, N-methionyl the human growth hormone; bovine growth hormone; parathyroid hormone; thyroxine; insulin; proinsulin; relaxin; prolactin; glycoprotein hormones such as follicle stimulating hormone (FSH), thyroid stimulating hormone (TSH), and luteinizing hormone (LH); hepatic growth factor; growth factor fibroblast; prolactin; placental lactogenic; mouse gonadotropin-associated peptide; inhibin; activin; vascular endothelial the tion growth factor (VEGF); integrin; thrombopoietin (TPO); nerve growth factors such as NGFβ; growth factor platelet; transforming growth factors (TGFs)such as TGFα and TGFβ; erythropoietin (EPO), interferon-type IFNα, IFNβ and IFNγ; colony-stimulating factors such as M-CSF, GM-CSF and G-CSF; interleukins type IL-1, IL-la, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12; TNFα or TNFβ. Preferred cytokines, according to the invention, the interferon and TNFα.

The term "cytotoxic agent" is used here to refer to a substance that inhibits or prevents the function of cells and/or causes destruction of cells. The term includes radioactive isotopes, chemotherapeutic agents, toxins, such as enzymatically active bacterial toxins, toxins, fungal, plant or animal origin, or fragments thereof. The term may also include members of the family of cytokines, preferably IFNγand antineoplastic agents with cytotoxic activity.

The term "chemotherapeutic agent" or "anti-cancer agent" is considered, according to the invention, as a member of the class "cytotoxic agents", as defined above, and includes chemicals that have antitumor activity, that is, prevent the development, maturation or proliferation of tumor cells, direct the military to a tumor cell, for example, with cytostatic or cytotoxic activity, and not indirectly through mechanisms such as the modification of the biological response. Suitable chemotherapeutic agents, according to the invention, preferably natural or synthetic chemical substance, but a biological molecule type proteins, polypeptides, etc. is not completely excluded. There are a large number of anticancer agents, available in clinical trials and pre-clinical development, which could be included in the present invention for the treatment of tumors/neoplasms method of combination therapy with TNFα and anti-angiogenic means, as cited above, optionally with other means, such as antagonists of EGF receptor. It should be noted that chemotherapeutic agents can be applied arbitrarily, together with the above combination. Examples of chemotherapeutic or products containing alkylating agents can be, for example, nitrogen mustards, etieleniminne derivatives, alkyl sulfonates and other compounds with an alkylating action, such nitrosomonas, cisplatin and dacarbazine; antimetabolites, such as folic acid, purine or pyrimidine antagonists; mitotic inhibitors, for example, Vinca alkaloids and derivatives of podophyllotoxin; cytotoxic the ski antibiotics and derivatives camptothecin. Preferred chemotherapeutic agents or chemotherapy involves amifostine (ethyol), cisplatin, dacarbazine (DTIC), dactinomycin, mechlorethamine (nitrogen mustard), streptozocin, cyclophosphamide, carmustin (BCNU), lomustin (CCNU), doxorubicin (adriamycin), doxorubicin lipo (doxil), gemcitabine (Gemzar), daunorubicin, daunorubicin lipo (daunoxome), procarbazine, mitomycin, cytarabine, etoposide, methotrexate, 5-fluorouracil (5-FU), vinblastine, vincristine, bleomycin, paclitaxel (Taxol), docetaxel (Taxotere), aldeslakin, asparaginase, busulfan, carboplatin, cladribine, camptothecin, CPT-11, 10-hydroxy-7-Etiketten (SN38), dacarbazine, floxuridine, fludarabine, hydroxyurea, ifosfamide, idarubitsin, mesna, alpha interferon, beta interferon, irinotecan, mitoxantrone, topotecan, leuprolide, megestrol, melphalan, mercaptopurine, plicamycin, mitotane, pegaspargase, pentostatin, pipobroman, plicamycin, streptozocin, tamoxifen, teniposide, testolactone, tioguanin, thiotepa, uracil mustard, vinorelbine, chlorambucil, and combinations thereof.

Preferred according to the invention, chemotherapeutic agents - cisplatin, gemcitabine, doxorubicin, paclitaxel (Taxol) and bleomycin.

The term "cancer" and "tumor" refers to or describes the physiological condition of mammals that is typically characterized by unregulated R is a shrub cells. Through the pharmaceutical compositions in accordance with the present invention, can be treated tumors of this type: tumors of the breast, heart, lung, small intestine, colon, spleen, kidney, bladder, head and neck, ovary, prostate, brain, pancreas, skin, bone, bone marrow, blood, thymus, uterus, ovary, larynx and liver. More specifically, the tumor is selected from the group consisting of adenoma, angiosarcoma, astrocytoma, epithelial cancer, germinoma, glioblastomas, gliomas, hamartomas, hemangioendothelioma, hemangiosarcoma, hematoma, hepatoblastoma, leukemia, lymphoma, Protocol, melanoma, neuroblastoma, osteogenic sarcoma, retinoblastoma, rhabdomyosarcoma, sarcoma and teratoid tumors. In particular, the tumor is selected from the group which includes lentigines limbs (acral lentiginous melanoma), actinic keratoses, adenocarcinoma, cystadenoma carcinoma, adenoma, adenosarcoma, adenosquamous carcinoma, astrocytic tumors, carcinoma of Bartolomeu cancer, basal cell carcinoma, carcinoma, bronchial gland, capillary, carcinoids, carcinoma, carcinosarcoma, cavernous/cholangiocarcinoma, chondrosarcoma, papilloma/carcinoma holodnogo plexus carcinoma bright cells, cystic adenoma, the tumor endodermal sinus, endometrial hyperplasia, endometrial stromal RAA the WMD, endometrioid adenocarcinoma, ependymoma, epithelioma, Ewing sarcoma, fibrolamellar carcinoma, Central nodular hyperplasia, gastrinoma, tumor germ cells, glioblastoma, glucagonoma, hemangioblastoma, hemangioendothelioma, hemangioma, hepatic adenoma, hepatic adenomatosis, hepatocellular cancer, insulinoma, intraepithelial the neoplasia, bezepitelialnye squamous cell neoplasia, invasive squamous cell carcinoma, large cell carcinoma, leiomyosarcoma, lentigo, maligna melanoma, malignant melanoma, malignant mesothelial tumor, medulloblastoma, medulloepithelioma, melanoma, meningeal, mesothelial metastatic cancer, mucoepidermoid cancer, neuroblastoma, neuroepithelial adenocarcinoma, nodular melanoma, oat cell carcinoma, oligodendroglioma, osteogenic sarcoma, pancreatic polypeptide, papillary serous adenocarcinoma, the tumor cells of the pineal gland, pituitary tumors, plasmacytoma, pseudosarcoma, pulmonary blastoma, renal cancer cells, retinoblastoma, rhabdomyo-sarcoma, sarcoma, serous cancer, small cell carcinoma, carcinoma of the soft tissue, somatostatin-secreting tumor, squamous cancer, squamous cell carcinoma, pedestalling superficial spreading melanoma, undifferentiated carcinoma, Avelino is melanoma, warty cancer, vipoma, well-differentiated tumors and tumor Wilma.

Pharmaceutical compositions of the invention may include tools that reduce or eliminate adverse factors associated with combination therapy of the present invention (the"secondary treatment"), including, but not limited to those means, for example, which reduces the toxic effect of anticancer drugs, such as inhibitors of bone absorption, cardioprotective funds. The mentioned extension means to prevent or reduce the side effects, nausea and vomiting associated with chemotherapy, radiotherapy or surgery, or reduce the effect of infection associated with the appointment mielosupressiveh anti-cancer drugs. Incremental means known from the prior art. Immunotherapy, according to the invention can optionally be administered with stimulants, such as BCG and immunostimulatory. In addition, the composition may include immunotherapeutics funds or chemotherapeutic agents, which contain cytotoxic effective Radiometrie isotopes or other cytotoxic agents such as cytotoxic peptides (e.g., cytokines or cytotoxic drugs, etc.

The term "pharmaceutical kit for treating tumors or tumor metastasis is relates to packaging and, as a rule, instructions use of reagents in methods of treatment of tumors and tumor metastases. The reagent in the kit of this invention is usually formulated as a therapeutic composition as described herein, and can therefore be in any of various forms, suitable for placement in the set. Such forms may include liquid, powder, tablet, suspension and a similar composition to ensure antagonist and/or fusion protein of the present invention. The reagents may be in separate containers suitable for the purpose, separately, according to this method, or alternatively can be combined in the composition in a separate container in the package. The package may contain a quantity sufficient for one or more reagent dosages, according to methods of treatment described herein. The kit of this invention also contains "instructions for use" of the materials contained in the package.

The term "pharmaceutical treatment" refers to therapeutic methods of the present invention, for the treatment of tumor cells in tumors and tumor metastasis, based on combined therapy inhibition of angiogenesis (anti-angiogenesis and anti-tumor immunotherapy using blocking means receptor to tyrosinekinase, preferably ErbB antagonists, primarily anti-ErbB1 (EGFR, Her1)/anti-rbB2 (Her2) antibodies. More than one media type inhibition of angiogenesis can be used in combination with more than one type, preferably, the inhibiting means anti-ErbB receptor. Sharing can be performed simultaneously, sequentially, or with the time interval between treatments. Any special therapy can be applied more than once during the course of treatment. The method can lead to synergistic potentiating effect of inhibiting the proliferation of tumor cells of each individual therapy, giving a more effective treatment than found by using only the individual component. Thus, in one aspect, the method of the invention comprises applying to the patient, in combination, the amount of anti-angiogenic means and the anti-ErbB receptor (Her1/Her2), which may not result in effective inhibition of angiogenesis, or antitumor cell activity, given that the number of individually. The method of the invention encompasses a variety of models for practicing the invention Paladino. For example, the means according to the invention, can be applied simultaneously, sequentially, or separately. In addition, the blocking means tyrosine kinase receptor, and anti-angiogenic agent can be applied separately at intervals of time of about 3 weeks between destinations is tions, i.e. mainly immediately after the first active drugs used until approximately 3 weeks after the first tool used. The method can be applied after the surgical procedure. Alternatively, a surgical procedure can be practiced in the interval between the appointment of the first active ingredient and the second active substance. A sample of this method is the combination of this method with surgical removal of the tumor. The treatment according to the method generally includes therapeutic compositions with one or more cycles of destination. For example, when simultaneously appoint a therapeutic composition comprising both agents used in the time interval approximately from 2 days to 3 weeks in a separate loop. After the cycle can be repeated, if necessary, according to the doctor. Similarly, when it is assumed sequential application, the destination time for each individual therapy should be adjusted to close the same interval of time. The interval between cycles can vary from zero up to 2 months. Monoclonal antibodies, polypeptides or organic mimetics/chemoterapeutic of this invention can be administered parenterally by injection or infusion after some time. Despite the fact that h is about the fabric want to treat, can usually be achieved in the body by way of General application and therefore most often treated by intravenous administration of therapeutic compositions, other tissues and delivery vehicles to consider, where there is a likelihood that the target tissue contains the target molecule. Thus, monoclonal antibodies, polypeptides or organic methods of this invention can be applied vnutriglazna, intravenously, intraperitoneally, intramuscularly, subcutaneously, vnutripolostno, transdermal, orthotopic injections and infusions, and can also be fitted with a peristaltic means. Therapeutic compositions containing, for example, an integrin antagonist, of this invention, traditionally used intravenously, such as by injection of a single dose. Therapeutic compositions of this invention contain a physiologically acceptable carrier together with an appropriate tool, as described herein, dissolved or dispergirovannom as the active ingredient.

As used herein, the term "pharmaceutically acceptable" and its grammatical variations, as he refers to compositions, carriers, diluents and reagents, are used equally, and is that matter is permissible to use mammal without showing undesirable Fiziol the environmental consequences such as nausea, dizziness, gastric upset and the like, the Preparation of a pharmaceutical composition that contains active ingredients that are dissolved or dispergirovannom a form that is understandable in technology and should not be restricted on the basis of the composition. Typically, such compositions are prepared as injectable or as liquid solutions or suspensions, however, solid forms suitable for solution in, or suspension, can also be prepared in the form of a solution before use. The preparation can also be in the form of an emulsion. The active ingredient may be mixed with excipients that are pharmaceutically acceptable and compatible with the active ingredient and in amounts suitable for use in therapeutic methods described herein. Fit excipient, for example, water, saline, dextrose, glycerol, ethanol or the like, and combinations thereof. In addition, if necessary, the composition may contain minor amounts of auxiliary substances such as wetting or emulsifying agents, buffer funds, etc. that enhance the effectiveness of the active ingredient. A therapeutic composition of the present invention can include pharmaceutically acceptable salts of the components. Pharmaceutically acceptable salts include salts of joining acids (formed by the s free amino groups of the polypeptide), derived from mineral acids such as, for example, hydrochloric or phosphoric acids, or with organic acids such as acetic, tartaric, almond, etc. Salts formed free carboxyl groups can also be derived from inorganic bases, such as, for example, hydroxides of sodium, potassium, ammonium, calcium or iron, and such organic bases as Isopropylamine, trimethylamine, 2-ethylaminoethanol, histidine, procaine and the like, Especially preferred HCl salt used in the preparation of cyclic polypeptide αvantagonists. Physiologically acceptable carriers known in the prior art. Examples of liquid carriers are sterile aqueous solutions that do not contain further substances in addition to the active ingredients and water, or contain a buffer such as sodium phosphate with the physiological value of pH value, physiological saline or both phosphate buffer and saline. Next, aqueous carriers can contain more than one buffer salt, and salts such as sodium and potassium chloride, dextrose, polyethylene glycol or other solvents. Liquid compositions may also contain a liquid phase in addition to and excluding water. Examples of such additional liquid phases is glycerin, vegetable oils, such Okhlopkova oil, and water-oil emulsion.

Typically, a therapeutically effective amount of immunotherapy in the form of, for example, antibodies anti-ErbB receptor or antibody fragment or conjugate antibody, or antibody anti-angiogenic receptor, fragment or conjugate is a number used in a physiologically acceptable composition, is sufficient to achieve a plasma concentration of approximately from 0.01 micrograms (μ (g) per milliliter (ml) to about 100 μg/ml, preferably from about 1 μg/ml to approximately 5 μg/ml, and usually about 5 μg/ml, the Dosage may be assigned differently and can vary from about 0.1 mg/kg to about 300 mg/kg, preferably from about 0.2 mg/kg to about 200 mg/kg, most preferably from about 0.5 mg/kg to about 20 mg/kg, in one or more doses daily for one or several days. If immunotherapy means is in the form of a fragment of a monoclonal antibody or mates, the number can be adjusted in terms of the mass of the fragment or conjugate relative to the weight of the whole antibody. The preferred plasma concentration in molar concentration is approximately 2 micromol (μM) CA is approximately to 5 mmol (mm) and preferably, approximately 100 μM 1 mm antagonist antibodies. A therapeutically effective amount, according to the invention, which is neimanmarcus.com peptide or protein of the polypeptide (for example, IFN-α), or other small molecule of this size is usually such an amount of the polypeptide, the use of which in a physiologically acceptable composition, is sufficient to achieve a plasma concentration of approximately from 0.1 micrograms (μ (g) per milliliter (ml) to about 200 μg/ml, preferably, from about 1 μg/ml to approximately 150 μg/m1. For a polypeptide having a mass of approximately 500 grams per mole, the preferred plasma concentration in moles is approximately 2 micromol (μM) to about 5 mmol (mm) and preferably about 100 μM 1 mm antagonist polypeptide. A typical dosage of the active substance, which is, preferably, a chemical antagonist or (chemically) chemotherapeutic agent, according to the invention, no immunotherapy tool or deimante.samaviciute peptide/protein) - 10 mg - 1000 mg, preferably about 20-200 mg, and more preferably, 50 to 100 mg per kilogram of body weight per day.

The term "therapeutically effective" or "terapevticheskii number" refers to the quantity of a drug, effective to treat a disease or disorder in the mammal. In the case of cancer, a therapeutically effective quantity of a drug can reduce the number of cancer cells; reduce the tumor size; inhibit (i.e., to reduce to some extent and preferably stop) cancer cell infiltration into peripheral organs; inhibit (i.e., to reduce to some extent and preferably stop) tumor metastasis; inhibit, to some extent, tumor growth; and/or relieve to some extent one or more symptoms associated with cancer. Significantly, the drug may prevent growth and/or kill cancer cells, it may be cytostatic and/or cytotoxic. For cancer therapy, efficacy can, for example, be measured by assessing the time to progression (TTP) and/or determining responsiveness (RR).

Example: brief clinical report of tests:

Patient, 45 years, initially suffered from progressing to squamous cell cancer of the upper jaw.

EMD 72000 monoclonal antibody human 425 (h425), Merck KgaA, Germany

EMD 121974: Cyclo-(Arg-Gly-Asp-DPhe-NMeVal), Cilengitide®, Merck KgaA, Germany

Chemotherapy: varied (gemcitabine, cisplatin, and so on)

The patient's history and clinical results/status at the beginning of the use astragalin the treatment: In July 1997 the patient is first delivered in Virchow Klinikum, Germany. Biopsies were taken of the suspected large tumor in the upper jaw. Histology showed squamous cell carcinoma classified as T4 N0 M0. on August 5, 1997, was given a partial resection of the upper jaw and resection of regional lymph nodes. Histological studies showed that was not achieved net border, was made additional resection during the same hospital stay. Due to unfavorable histological classification patient received postoperative radiation therapy to 50.4 Gy with September and October 1997

In July 1998, was suspicious about the progression of the illness that led to hospitalization. Now histology showed adenosquamous carcinoma. After consulting doctors - radiologists was recommended by another radiation therapy, which began in August 1998, the Patient was simultaneously treated with gemcitabine (100 mg) as a sensitizer exposure. 6-week therapy resulted in complete clinical remission.

During the combined radio-chemotherapy the patient received treatment with 1000 mg gemcitabine (5 courses to 16 destinations).

In March 1999, again there was a progression of the cancer, which led to additional radiation therapy and palliative tumor resection. In August 1999, again prog is Essie tumor and was started on chemotherapy with cisplatin (75 mg/m 2) and docetaxel (75 mg/m2). After three appointments, therapy was stopped due to the lack of effect on tumor growth.

Diffuse bleeding from a large tumor mass required frequent transfusions of concentrated erythrocyte.

The compassionate course of treatment with anti-angiogenic tools / chemotherapeutic agents: After treatment with EMD 121974 (600 mg/m2) and gemcitabine (Gemzar) (1000 mg/m2in November 1999 he was diagnosed regression of the tumor. Since mid-January 2000, the patient was again able to hear in the right ear, and he was able to open his mouth to 30% more than in December 1999, the Surface of the tumor showed symptoms of granulation and healing of small wounds.

The bleeding stopped, and there was no need for further transfusions. The patient was treated EMD 121974 and Gemzar from 17.11.1999 on 30.03.2000. With 06.04.2000 on 28.04.2000 EMD 121974, Gemzar and chemotherapy with 5-FU, cisplatin and reschooling was attributed to the patient, because it was discovered the tumour progression. Chemotherapy treatment was stopped because of hematotoxicity and the treatment continued only cilengitide. From April to June 2000, the patient received 600 mg/m2EMD 121974 twice a week, which only resulted in stability of the disease.

The patient's condition deteriorated after a few weeks, and the patient began Leche is to be increased dose of 1200 mg/m 2EMD 121974 twice a week.

Treatment with h425 + cilengitide + chemotherapy: EMD 72000 first was given in November 2000 at a dosage of 200 mg (infusion within half an hour), after predlejania with dexamethasone / gemeindebauten (Fenistil) and ranitidine Zantic). After one week the patient received additional gemcitabine (1000 mg/m2).

Weekly treatment plan was as follows: Monday: 1200 mg/m2cilengitide (infusion within hours), Thursday 200 mg EMD 72000 (infusion within half an hour), the next 1000 mg/m2gemcitabine (infusion within hours), Friday 1200 mg/m2cilengitide (infusion for one hour). After such treatment was found createrootpane decomposition of the tumor mass. Tumor masses were surgically removed in a few cases. The doctors believed the effect of the combined treatment is extremely significant. No adverse reactions during therapy drugs EMD 121974 and EMD 72000 not been identified. Still the patient's condition remained superior.

1. The pharmaceutical composition intended for the treatment of tumors and tumor metastasis, including

(i) at least one antibody or a functionally intact derivative containing the binding site that binds to an epitope ErbBl(Herl) receptor and

(ii) at least one inhibitory Angie ESIS means, optionally together with a pharmaceutically acceptable carrier, diluent or recipient.

2. The pharmaceutical composition according to claim 1, in which the specified antibody is chosen from a group that includes humanitariannet monoclonal antibody 425 to EGFR and chimeric monoclonal antibody 225 (s) to EGFR.

3. The pharmaceutical composition according to claim 1, in which the specified inhibiting angiogenesis agent, is αvβ3that αvβ5or αvβ6inhibiting integrin tool or a means of blocking VEGR receptor.

4. The pharmaceutical composition according to claim 3, in which the specified inhibiting integrin tool is an RGD-containing linear or cyclic peptide, or antibody, including its intact derived.

5. The pharmaceutical composition according to claim 4, in which the specified RGD-containing peptide is cyclo-(Arg-Gly-Asp-DPhe-NMeVal).

6. The pharmaceutical composition according to claim 3, where the specified inhibiting the integrin agent is an antibody or its active derivative, selected from the group comprising LM609, P1F6 and E.

7. The pharmaceutical composition according to claim 1, which additionally comprises a cytotoxic agent.

8. The pharmaceutical composition according to claim 7, where the cytotoxic agent is selected from a group that includes cisplatin, doxorubicin, gemcitabine, docetaxel, pacl taxel, bleomycin.

9. The pharmaceutical kit for the treatment of tumors and tumor metastasis, including

(i) a first package containing at least one antibody or a functionally intact derivative, comprising a binding site that binds to an epitope ErbBl(Herl) receptor and

(ii) a second package containing at least one inhibiting angiogenesis agent.

10. The pharmaceutical kit according to claim 9, in which the specified antibody is chosen from a group that includes humanitariannet monoclonal antibody 425 to EGFR and chimeric monoclonal antibody 225 (s) to EGFR.

11. The pharmaceutical kit according to claim 9, in which the specified inhibiting angiogenesis agent, is αvβ3that αvβ5or αvβ6inhibiting integrin tool or a means of blocking VEGR receptor.

12. The pharmaceutical kit according to claim 11, in which the specified inhibiting integrin tool is an RGD-containing linear or cyclic peptide, or antibody, including its intact derived.

13. The pharmaceutical kit according to item 12, in which the specified RGD-containing peptide is cyclo-(Arg-Gly-Asp-DPhe-NMeVal).

14. The pharmaceutical kit according to claim 11, where the specified inhibiting the integrin agent is an antibody or its active derivative, vibrancies group, including LM609, P1F6 and E.

15. The pharmaceutical kit according to claim 9, which additionally a third package comprising a cytotoxic agent.

16. The pharmaceutical kit according to item 15, where the cytotoxic agent is selected from a group that includes cisplatin, doxorubicin, gemcitabine, docetaxel, paclitaxel, bleomycin.

17. A method of treating tumors or tumor metastases in a patient, which comprises administration to the patient a therapeutically effective amount or means including

i) at least one antibody or a functionally intact derivative, comprising a binding site that binds to an epitope ErbBl(Herl) receptor and

(ii) at least one inhibiting angiogenesis agent.

18. The method according to 17, in which the indicated antibody is chosen from a group that includes humanitariannet monoclonal antibody 425 (h425) to EGFR, a chimeric monoclonal antibody 225 (s) to EGFR.

19. The method according to 17, in which the specified means of inhibiting angiogenesis is αvβ3that αvβ5or αvβ6inhibiting integrin tool or a means of blocking VEGR receptor.

20. The method according to claim 19, in which the specified inhibiting integrin tool is an RGD-containing linear or cyclic peptide, or antibody, including its intactivists.

21. The method according to claim 20, wherein said RGD-containing peptide is cyclo-(Arg-Gly-Asp-DPhe-NMeVal).

22. The method according to 19, where the specified inhibiting the integrin agent is an antibody or its derivative selected from the group comprising L609, 1F6 and E.

23. The method according to claim 19, allowing for introduction to the patient a cytotoxic agent.

24. The method according to item 23, where the cytotoxic agent is selected from the group which consists of: cisplatin, doxorubicin, gemcitabine, docetaxel, paclitaxel, bleomycin.

25. The method according to 17, which includes an introduction to the patient a therapeutically effective amount of (i) gumanitarnogo monoclonal antibody 425 (h425) or chimeric monoclonal antibody 225 (s), (ii) cyclo-(Arg-Gly-Asp-DPhe-NMeVal), and optionally (iii) cisplatin, doxorubicin, gemcitabine, docetaxel, paclitaxel, bleomycin.



 

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SUBSTANCE: method involves dividing cells by using agglutination. Required reagents comprise dextrane solution, anti-Glycophorin A-antibody, anti-CD15-antibody, anti-CD9-antibody, and antibodies selected from a group composed of anti-CD3-antibody, anti-CD72-antibody, anti-CD16-antibody, anti-CD4-antibody, anti-CD8-antibody and anti-CD2-antibody.

EFFECT: enhanced effectiveness in selecting blood cells of rare cell types with high output amount.

33 cl, 17 tbl

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EFFECT: high accuracy of diagnosis.

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EFFECT: high accuracy of prognosis.

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1 ex

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1 tbl

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1 tbl

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EFFECT: valuable medicinal property of preparation.

1 tbl, 2 ex

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4 cl, 1 tbl, 3 ex

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SUBSTANCE: invention describes a medicament used in treatment of liver disease by prevention of invasion onset of portal vein. This agent represents the effective medicament containing menatetrenone as an active component used in treatment/prophylaxis of liver disease. Indicated agent used in treatment/prophylaxis of liver disease is the effective medicament against liver cancer, in particular, against DCP (des-γ-carboxyprothrombin)-positive liver cancer. Agent used in aims for treatment/prophylaxis of liver disease contains menatetrenone as an active component, it improves prognosis significantly after carrying out the anticancer therapy and possesses the excellent effect directed on prevention of liver cancer relapses.

EFFECT: valuable medicinal property of agent.

4 cl, 2 tbl, 10 dwg, 2 ex

FIELD: medicine.

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79 cl, 28 dwg, 44 tbl

FIELD: organic chemistry, medicine, pharmacy.

SUBSTANCE: invention relates to novel derivatives of pyridylcyanoguanidines of the general formula (I):

or their pharmaceutically acceptable salts wherein 1 means hydrogen atom; X means hydrocarbon (C1-C12)-biradical; Y means a bond or oxygen atom (O); Z means 5-10-membered aromatic heterocyclic radical optionally substituted with hydroxy-group under condition that R1 is not bound to nitrogen atom in pyridyl ring. Compounds of the formula (I) and their salts possess antiproliferative activity and can be used in medicine.

EFFECT: valuable medicinal properties of compounds and pharmaceutical composition.

9 cl, 1 tbl, 7 ex

FIELD: medicine.

SUBSTANCE: compound is represented by structural formula

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19 cl, 90 tbl

Kahalalide f // 2292216

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SUBSTANCE: invention relates to novel compositions and using kahalalide F, to a set containing the kahalalide F composition and to a reduced solution prepared from the kahalalide F composition. Combination of non-ionic surface-active substance and organic acid is suitable for using with a filling agent for preparing lyophilized formulation of kahalalide F.

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10 cl, 7 ex

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2 ex, 3 tbl

FIELD: medicine.

SUBSTANCE: preparation belongs to gestagen class as 17α-acetoxy-3β-butanoyloxy-6-methyl-pregna-4,6-dien-20-on allowing peroral application. The compound shows no toxic activity and has no androgenic side effect. It is found to be better than Depo-Provera gestagen used in clinical practice and possesses chemosensitizing activity. The preparation is obtained by megestrol acetate reduction with sodium borohydride with following product etherification with butyric acid anhydride. The reaction mixture is treated with ammonia solution after etherification with butyric acid excess being removed to increase output and target product separation.

EFFECT: enhanced effectiveness of treatment.

3 cl, 3dwg, 1 tbl

FIELD: medicine.

SUBSTANCE: method involves applying chlorocholine chloride as antitumor means.

EFFECT: enhanced effectiveness of treatment; reduced risk of adverse side effects manifestation; low toxicity.

FIELD: veterinary science.

SUBSTANCE: the suggested preparation contains thiamuline, polymixin or antibiotic of tetracycline group and, also, a target additive at the following content of the above-mentioned components, weight%: thiamuline 0.25-90.0, polymixin or antibiotic of tetracycline group 2.5-90.0%, a target additive - the rest. The preparation in question is of high prophylactic and curative efficiency and wider spectrum of antimicrobial action.

EFFECT: higher efficiency of prophylaxis and therapy.

3 cl, 14 ex, 11 tbl

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