JP2008502588A - Laminin-5 modulator and use thereof - Google Patents

Abstract

          Disclosed herein are methods for treating or preventing various diseases and disorders using modulators of laminin-5. Typical diseases and disorders are inflammatory bowel diseases including ulcerative colitis and Crohn's disease, as well as multiple cystic kidneys and cancers associated with one or more of the aforementioned symptoms. Exemplary laminin-5 modulators include monoclonal antibodies against laminin-5 γ2 chain and small interfering RNA (siRNA) sequences against nucleotide sequences encoding laminin-5 γ2 chain. Combination therapy based on the administration of laminin-5 modulators and at least one other therapeutic agent is also described.

Description

          The present invention relates to methods and compositions for the treatment and / or prevention of diseases or disorders associated with altered laminin-5 expression and / or function.

          Inflammatory bowel disease (or “IBD”) is a disease characterized by inflammation of the gastrointestinal (“GI”) tract that is not usually caused by other factors such as infection, medication or malignancy. IBD includes at least two related symptoms: ulcerative colitis (UC) and Crohn's disease (CD). UC typically affects only the colon, and in some cases can be treated by surgical removal of the colon (colectomy). Many CD patients also require surgery at least once during their lifetime. However, CDs are currently not generally considered to be curable. Studies in this area have demonstrated that many CD and UC patients have an increased risk of developing colorectal cancer. Colorectal cancer is the leading cause of death in North America and Europe.

          For these and other reasons, there remains a need for alternative and improved methods of promoting, enhancing and / or inducing treatment and / or prevention of IBD and IBD-related cancers. The invention described herein includes such methods and novel compositions useful in the practice of such methods, as well as others including irritable bowel syndrome, polycystic kidney disease (PKD) and non-IBD associated cancers. Methods of inducing, promoting and / or enhancing the treatment and / or prevention of other diseases and disorders. These and other advantages of the invention, as well as further aspects and features of the invention, will be apparent from the description of the invention provided herein.

          The present invention employs various modulators of laminin-5 such as, for example, IBD, IBD-related cancers, irritable bowel syndrome (“IBS”) and various PKD caused by, for example, autosomal polycystic kidney disease (ADPKD). Various methods of inducing physiological responses associated with the treatment and / or prevention of diseases and disorders are provided. Also described herein is a combination method, ie, a method of treating the aforementioned diseases and disorders and cancer using a combination of a laminin-5 modulator and a second agent.

          Thus, in one embodiment, the present invention treats inflammatory bowel disease comprising administering to a subject suffering from inflammatory bowel disease a composition comprising an effective amount of a laminin-5 modulator. Provide a method. In another embodiment, the present invention relates to colorectal disease associated with inflammatory bowel disease, comprising administering to a subject suffering from inflammatory bowel disease a composition comprising an effective amount of a laminin-5 modulator. A method for reducing the risk of cancer is provided. In one aspect, the inflammatory bowel disease is ulcerative colitis. In the second aspect, the inflammatory bowel disease is Crohn's disease.

          The present invention also provides a method of treating polycystic kidney, comprising administering a composition comprising an effective amount of a laminin-5 modulator to a subject identified as suffering from polycystic kidney disease. . In another embodiment, the invention involves administering a composition comprising an effective amount of a laminin-5 modulator to a subject identified as suffering from autosomal dominant polycystic kidney disease. A method for inhibiting the progression of cystic kidney is provided.

          The invention also provides a method of treating cancer, comprising administering to a subject suffering from cancer a composition comprising an effective amount of a laminin-5 modulator and at least one second anticancer agent.

          In certain aspects, in any of the above methods, the laminin-5 modulator is an antibody, laminin-5 binding protein, siRNA, antisense molecule or small molecule Ln-5 modulator directed against laminin-5. Selected from the group consisting of In another aspect, the method of claim 8, wherein the laminin-5 modulator is an antibody directed against a laminin-5 γ2 chain. In yet another aspect, the laminin-5-modulator is a siRNA derived against a nucleic acid encoding a laminin-5 γ2 chain.

          The present invention also provides the use of laminin-5-modulators in the preparation of a medicament for treating inflammatory bowel disease. In one aspect, the inflammatory bowel disease is ulcerative colitis. In another aspect, the inflammatory bowel disease is Crohn's disease.

          The present invention also provides the use of laminin-5-modulators in the preparation of a medicament for treating inflammatory bowel disease-related colorectal cancer.

          The invention also provides the use of an effective amount of laminin-5-modulator and at least one second anticancer agent in the preparation of a medicament for treating cancer.

          The present invention also provides the use of an effective amount of laminin-5-modulator and at least one second anticancer agent in the preparation of a medicament for reducing tumor growth.

          In certain aspects, in any of the above uses, the laminin-5 modulator is an antibody, laminin-5 binding protein, siRNA, antisense molecule or small molecule Ln-5 modulator directed against laminin-5 Selected from the group consisting of In one aspect, the laminin-5 modulator is an antibody directed against laminin-5 γ2 chain. In another aspect, the laminin-5-modulator is an siRNA derived against a nucleic acid encoding a laminin-5 γ2 chain.

          These aspects of the invention and various further aspects are described elsewhere herein.

          The invention described herein is in part, for example, PKD caused by, for example, IBD, IBD-related cancers, irritable bowel syndrome (“IBS”), and autosomal polycystic kidney disease (ADPKD), for example. The use of laminin-5 (Ln-5) modulators to promote, enhance and / or induce physiological responses in connection with the treatment and / or prevention of various diseases and disorders such as Based on the discovery that it can. In one embodiment, the modulator of laminin-5 promotes a physiological response in connection with the treatment and / or prevention of diseases and disorders not previously known to be associated with laminin-5 modulation, Used to enhance and / or induce. One or more functions of Ln-5 and / or one or more functions of one or more biomolecules associated with Ln-5 (in an applicable patient or host in a normal and / or associated medical condition) Treatment and / or prevention of these and other diseases and disorders by administering Ln-5 modulators under conditions that are at least partially (eg, at least substantially or substantially completely) ineffective. The method of facilitating is another important feature of the present invention. For example, administration of an Ln-5 modulator to an IBD patient at risk of developing colorectal cancer can reduce the risk of cancer and / or restore the IBD pathology to milder symptoms. . In addition, Ln-5 modulators can slow or inhibit the progression of ADPKD, and combination therapy based on Ln-5 modulators and a second anticancer agent is used to treat cancer. It is possible.

          The invention described herein generally also provides novel compositions comprising one or more Ln-5 modulators that can be used in such methods and in the prevention and / or treatment of cancer. (Related methods are also provided.) Further important general aspects of the present invention include identifying Ln-5 modulators, preparing such compositions, and in connection with certain therapeutic, prophylactic and / or diagnostic applications. 5 Methods including, but not limited to, promoting the sale of preparative material compositions.

          As described herein, Ln-5 modulation can be achieved using n-5 modulators such as, for example, antibodies to Ln-5 and antisense technology. Ln-5 antibodies are described, for example, in U.S. Patent Nos. 5,660,982 and 6,143,505, U.S. Patent Application Nos. 2004/0120959, 2003/0100529 and 2002/0052307, European Patent No. 0 784 703, Ln-5 antisense nucleic acids and other related molecules and methods are described, for example, in US Patent Application No. 20030224993 and “Seftor et al., Cancer Res. 2001 Sep 1; 61 (17). ): 6322-7 ”.

          Laminin-5 (“Ln-5”) is a basement membrane extracellular matrix macromolecule that provides an adherent substrate for both adhesion and migration in a variety of cell types. Structurally, Ln-5, like other laminins, is composed of α, β and γ chain subunits (Ln-5 specifically has the general structure α3β3γ2). Ln-5 is expressed primarily in the epithelial basement membrane, and the biological functions of Ln-5 in the epithelial basement membrane include cell scaffolding and migration. Ln-5 γ2 chain expression is found at the forefront of invasion of tumors such as colorectal cancer, as well as in UC and ADPKD tissue samples (Habermann et al., Scand J Gastroenterol 2001; 36: 751-8 and Joly et al., Am J Pathol 2003; 163: 1791-1800).

          As shown in Example 1 and FIGS. 1 and 2, in an animal model of IBD, Ln-5 γ2-expression in tissues affected by IBD is observed in crypt cells and a portion of crypts. It was. Example 2 reports that in vitro Ln-5 modulation based on siRNA administration successfully reduced the expression of the Ln-5 γ2 chain, as shown in FIG. Example 3 shows that the use of siRNA on the Ln-5 γ2 chain increases spontaneous cell death, and that increased cell death is retained when siRNA is administered in combination with other anticancer compounds. It is shown that.

Ln-5 modulator
In general, the methods of the present invention may be any suitable substance that modulates the biological activity of Ln-5 (“Ln-5 modulator”) in a manner suitable to achieve the desired physiological result. ). In certain embodiments, a combination of two or more different Ln-5 modulators is combined to achieve the desired physiological result.

          As described herein, “Ln-5 modulation” includes reduction of Ln-5 expression, reduction of binding of Ln-5 and Ln-5 ligands and / or of cells expressing Ln-5. This includes, but is not limited to, a reduction in in vivo or in vitro proliferative capacity or migration or invasion capacity, or any combination of the foregoing. Ln-5 ligands and Ln-5 binding proteins that may serve as Ln-5 modulators are exemplified below.

          Ln-5 modulators include, for example, anti-Ln-5 antibodies (“Ln-5 antibodies”) (in normal and / or disease states in the target host) and other peptides that interact with Ln-5 An antibody that binds to a protein and / or polypeptide. Such non-immunoglobulin Ln-5 associated polypeptides, peptides and proteins can be referred to as “Ln-5 binding proteins” (the terms polypeptide, peptide and protein are not otherwise described or apparently Unless the context contradicts, it is used interchangeably throughout this specification and derivatives of such polypeptides (for example, international patent applications WO 02/083851, WO 03/048185 and WO 03/102166 and among them) Glycosides, polypeptides modified by acylation, etc.) as described in the relevant references cited in Exemplary anti-LN-5 antibodies include 5D5 as described in International Patent Application No. WO 2004/039401 and US Patent Application No. 2004/0120959, which are hereby incorporated by reference in their entirety. And 6C12. Collectively, such anti-Ln-5 antibodies and antibodies to Ln-5 binding proteins can be referred to as Ln-5 related antibodies.

          Additional Ln-5 modulators include functional nucleic acid molecules (eg, Ln-5) such as antisense nucleic acids (typically oligonucleotides) derived against polypeptides that modulate Ln-5 activity in vivo. -5 itself, as well as polypeptides that modulate Ln-5 in relation to cancer progression, IBD development, ADPKD development, etc.-as further described herein, examples include Certain integrins, MMP-2 and / or BMP-1 may be included); proteins and / or nucleic acid aptamers (including intramers—for example, “Famulok et al., Chem Biol. 2001 Oct; 8 (10 ): 931-9 ”)), typically interfering RNA molecules that are double stranded (ds) (eg, small or short interfering RNA ( siRNA ") molecules), and / or larger dsRNA molecules, ribozymes, triplex-forming molecules, foreign guide sequences and other" gene repressions "known in the art designed to be RISC cleaved into appropriate siRNAs. "Agent". A functional nucleic acid molecule is a specific that is carried by the target molecule such that Ln-5 activity is modulated, or that a functional nucleic acid molecule can possess nascent activity independent of any other molecule. It can act as an influencing factor, inhibitor, modulator and stimulator of the activity.

          In other aspects, the Ln-5 modulator is a demethylating agent (see, for example, “Urburadka et al., Clinical Cancer Research Vol. 9, 2665-2672, July 2003”), a gene activator (eg, Specific types of Ln-5 molecules, such as Ln-5 or Ln-5-related protein expression promoters and / or enhancers integrated into the genome of the target cell by homologous recombination techniques, and / or It can be a substance that up-regulates the production of Ln-5 binding protein. Ln-5 modulators are Ln-5 modulatory “small molecule” compounds (small molecules are generally organic compounds having a molecular weight of less than about 1,000 daltons, more typically less than about 500 daltons). Conceivable and typically exclude nucleic acids and peptides (especially of any significant length), but include sugars, individual amino acids, lipids, vitamins and non-peptide hormones such as, for example, Ln-5 linked heparin Can be included). Typically, small molecule Ln-5 modulators bind to Ln-5 and Ln-5 binding proteins under physiological conditions associated with an associated disease or pre-disease state in a subject, and Ln-5 It will disrupt the interaction between Ln-5 binding proteins.

Ln-5-associated antibody refers to an immunoglobulin molecule having the ability to specifically bind Ln-5, its components (for example, α3, β3, or γ2 chain, or their domain-domain 2 of γ2 chain, etc.) Fragment or derivative, ie Ln-5 binding protein. Unless otherwise stated or clearly contradicted by context, this definition of an antibody includes antibody fragments that retain the ability to specifically bind to Ln-5 or a portion thereof. Examples of binding fragments encompassed by the term “antibody” include, but are not limited to: (I) Fab fragment, monovalent fragment consisting of VL, VH, CL and CH I domains; (ii) F (ab) ₂ and F (ab ′) ₂ fragments, _two linked by disulfide bridges in the hinge region (Iii) an Fd fragment consisting of VH and CH1 domains; (iv) one amino acid residue of each of VH and VL is a cysteine residue via a disulfide bond between cysteine residues. DsFv fragment obtained by ligating substituted polypeptides (Reiter et al. (See Protein Engineering, 7, 697 (1994))); (v) dAb fragment consisting of VH domain (eg, Ward et al., (1989) Nature 341: 544-546); (vi) isolated phase A linear antibody (eg, Zapata et al. Protein Eng. 8 (10): 1057-1062 (1995) comprising a sex determining region (CDR) and (viii) a pair of tandem Fd segments forming a pair of antigen binding regions. In addition, although the two domains VL and VH of the Fv fragment are typically encoded by separate genes, they can be paired with the VL and VH regions using recombinant methods. Produced as a single protein chain, known as a monovalent molecule (single chain Fv (scFv); see, for example, Bird et al. (1988) Science 242: 423-426: and Huston et al. (1988) Proc Natl. Acad. Sci. USA 85: 5879-5883).) Can be linked by a synthetic linker that can be made to form. Such single chain antibodies are also encompassed within the term “antibody”. Unless otherwise stated, other forms of single chain antibodies such as diabodies are also encompassed by the term “antibody”. A diabody is a VH and VL domain expressed on a single polypeptide chain, but separated by a linker that is too short to allow pairing between two domains on the same chain. Are bivalent and bispecific antibodies that force the domain to pair with the complementary domain of another chain, creating two antigen-binding sites (eg, Holliger, P., et al. (1993) Proc. Natl. Acad. Sci. USA 90: 6444-6448; Poljak, RJ, et al. (1994) Structure 2: 1121-1123, EP 404,097 and WO 93/11161)).

          Also contemplated are “fully human antibodies” which are antibodies having both variable and constant regions derived from human germline immunoglobulin sequences. Such antibodies can be obtained from humanized transgenic animals (eg, Green et al.) That contain human immunoglobulin loci and unique immunoglobulin gene deletions, such as XenoMouse® (Abgenix-Fremont, CA, USA). al. Nature Genetics 7: 13-21 (1994); Mendez et al. Nature Genetics 15: 146-156 (1997); Green and Jakobovits J. Exp. Med. 188: 483-495 (1998); 463 151 B1; International patent applications WO 94/02602, WO 96/34096; WO 98/24893, WO 99/45031, WO 99/53049, and WO 00/037504; U.S. Patent Nos. 5,916,771, 5,939,598, 5,985,615 No. 5,998,209, 5,994,619, 6,075,181, 6,091,001, 6,114,598 and 6,130,364) or a transgenic vertebrate containing a minilocus of the human Ig coding gene is there.

          In certain aspects, the Ln-5 associated antibodies used in the various methods of the invention are “humanized antibodies”. A humanized antibody is a recombinant antibody in which complementarity determining region (CDR) sequences or variants thereof obtained from the germline of another mammalian species, such as a mouse, are grafted into a human “framework” sequence. . In one aspect of the invention, the isolated humanized monoclonal antibody comprises a CDR amino acid sequence derived from a monoclonal antibody. In a further aspect of the invention, the isolated humanized monoclonal antibody comprises a CDR amino acid sequence derived from a mouse monoclonal antibody.

          In one aspect, the Ln-5 associated antibody used in the various methods of the invention is a chimeric antibody, which is a molecule specific such that the molecule has reduced immunogenicity in humans. It refers to an antibody that has been altered in part. Such antibodies and methods for making them are well known in the art.

          The antibodies used in the various methods of the invention can be of natural or synthetic origin. Natural antibodies can be monoclonal or polyclonal antibodies, both of which are well known in the art. For synthetic antibodies, the antibody can be a non-natural immunoglobulin molecule that is altered by one or more amino acid substitutions, exchanges, additions and / or deletions, provided that such an immunoglobulin molecule is Ln-5, It shall retain the ability to bind Ln-5 binding protein or any portion thereof. Synthetic antibodies can also be “derivatives” of Ln-5 associated antibodies that have been altered by the addition of modified amino acids, chemical moieties, and the like. Unless otherwise stated, various methods of the invention involving Ln-5 associated antibodies can be performed using different chemical compositions, antigen specificities or combinations of both Ln-5 associated antibodies. is there.

          Typically, the Ln-5 associated antibody used in the method of the present invention is an antibody against Ln-5. In such methods, any suitable Ln-5 antibody can be used. Thus, for example, the methods of the present invention may include Ln-5 antibody that detectably binds the γ2 chain of Ln-5, Ln-5 antibody that detectably binds the α3 chain of Ln-5, or β3 of Ln-5 This can be done using an Ln-5 antibody that detectably binds the chain.

          Ln-5 binding protein refers to a non-immunoglobulin polypeptide that detectably binds Ln-5 under physiological conditions of normal and / or disease states. Examples of Ln-5 binding proteins include type VII collagen, fibulin-1, fibrin-2, BP180, syndecan-4, dystroglycan, nidogen-1, matrix metalloproteinases (eg, MMP-1, MMP-2, MMP -9, MMP-10 (stromelysin-2) and MMP-14 (also known as MT1-MMP)), matrix metalloproteinase tissue inhibitor-1 (TIMP-1) and TIMP-2, laminin-6, Laminin-7, EGF-R, Rho GTPase, phosphorylated hsp-27, p300, cytokeratin, E-cadherin, 67 kDa laminin receptor and bone morphogenetic protein 1 (BMP-1). In one aspect, the Ln-5 binding protein is an Ln-5 binding integrin. In such an aspect, the Ln-5 binding protein is a group consisting of, for example, alpha3beta1 (α3β1), alpha6beta1 (α6β1), alpha2beta1 (α2β1) and alpha6beta4 (α6β4) integrin. It is possible to select from.

          Ln-5 binding proteins are fragments of such proteins that retain Ln-5 binding ability or synthetic variants (homologs) of such proteins that retain Ln-5 binding ability or derivatives of such proteins or variants It can also be. Retention of Ln-5 binding ability is achieved by immobilizing a protein presumed to be Ln-5 binding protein on the surface, and after washing, soluble Ln− at stepwise concentrations (eg, 0.1 ng, 10 ng, 100 ng, 1 μg, 10 μg). Anti-Ln-5 antibody or anti-Ln-5 conjugated to fluorophore after incubation at room temperature (20 ° C.) for 1-24 hours before adding 5 and washing with standard ELISA wash buffer Laminin-5 is detected by using antibody alone, followed by addition of secondary anti-mouse antibody, using appropriate detection such as horseradish peroxidase, washing to ensure specificity, and finally detection By doing so, it is possible to detect by ELISA assay. The binding between Ln-5 and a candidate Ln-5 binding protein includes a protein (for example, an integrin heterodimer) that is membrane-bound in the liposome, and after the radioactivity is loaded inside the liposome, specific binding occurs. Can be detected by detecting the radioactivity bound to the surface coated with Ln-5 (10 nM) or laminin 10/11 (10 nM). Such methods are described in further detail in Nishiuchi et al. (J. Biochem. 2003; 134: 497-504). Candidate binding of Ln-5 and Ln-5 binding proteins may include Ln-5 and / or Ln-5 binding proteins (to detect cells that are stained during binding interactions, It is also possible to detect Ln-5 itself by using FACS analysis of cell lines expressing FITC or another fluorophore. Non-specific signals can be determined using staining of control cells lacking expression of Ln-5 or Ln-5 binding protein (proteins not stained need to be missing). The disruption of binding of Ln-5 and Ln-5 binding proteins may be determined by a reduction or absence of specific binding interactions as determined by the methods in the art as exemplified (but not limited to) by the above list. Is possible. As defined herein, an appropriate control (eg, the non-specific binding test described above, or the same assay, but omitting Ln-5 or irrelevant control instead of Ln-5) Ln-5 binding protein is a candidate for a Ln-5 binding protein that has a higher detection value in such an assay. Unless otherwise stated, it may be acceptable to use such combinations of Ln-5 binding proteins of the invention in the practice of the methods of the invention. An example may be a combination of the extracellular domains of α and β integrin subunits in such binding assays.

          In another aspect, the Ln-5 associated antibody is an antibody against an n-5 binding protein. Thus, for example, Ln-5 associated proteins include an antibody that binds to the β1 domain of the β subunit or an antibody that binds to the α1 domain / β propeller in the α subunit of the integrin, α2β1, α3β1, α6β1, or α6β4. It can be any antibody against Ln-5 binding integrin, such as an integrin. See also "Xiao et al., Nature (2004) vol 432 page 59-67". In another aspect, the Ln-5 associated antibody can be an antibody against MMP-2 or an antibody against BMP-1.

          The peptide Ln-5 modulator can be administered directly or can be expressed in the host by any suitable gene expression system. Thus, for example, the various methods of the present invention, upon expression, can include Ln-5 binding proteins (eg, wild type Ln-5 binding proteins or Ln-5 binding variants of such proteins, and / or among them). Any fragment or fusion protein comprising any of them) and / or by delivery of nucleic acids resulting in the production of Ln-5 associated antibodies. Such nucleic acids can be linear expression elements, plasmids, cosmids, or adenoviral vectors (eg, wild type or modified adenovirus-replication deficient and targeted adenoviruses), poxvirus vectors, herpes virus vectors. Can be advantageously delivered by a suitable vector, such as a viral vector such as a lentiviral vector, adeno-associated virus (AAV). As already mentioned, expression of such peptides can also be achieved by administration of factors that up-regulate such expression in terms of transduction factors, gene activation or other suitable techniques for such factors. Is possible.

The antisense Ln-5 modulator can be any suitable type of antisense nucleic acid molecule. A “classical” antisense molecule is a short molecule that typically includes a gene-specific sequence that suppresses translation of a target RNA by targeting a specific RNA transcript. Often, such molecules are modified such that the complex formed between the target transcript and the antisense molecule is digested by RNase H or a similar enzyme, thereby increasing the efficiency of the antisense molecule. . Other antisense molecules are engineered so that ribozymes (eg, RNaseP) can destroy target RNAs (eg, antisense molecules are so-called “foreign guide sequences” (EGS; (External Guide Sequences)). ). Ln-5 modulators can also be triple helix-forming nucleic acids, typically gene specific DNA molecules (whether such molecules are properly classified as antisense molecules are currently , Has become the subject of academic discussion.) These molecules can target genes, eg, genes encoding Ln-5 associated polypeptides that modify Ln-5 associated with certain stages of cancer progression and / or IBD development. . Antisense molecules can be designed to interact with the target nucleic acid molecule through either standard or non-standard base pairing. The interaction of a functional nucleic acid molecule and a target molecule is designed to facilitate the destruction of the target molecule, for example, through RNAse DNA degradation mediated by RNAseH. Alternatively, functional nucleic acid molecules are designed to interfere with processing functions such as transcription or replication that are naturally expected to occur on the target molecule. Antisense modulators can be designed based on the sequence of the target molecule. There are many ways to optimize the efficiency of binding by finding the most accessible region of the target molecule. Typical methods would be in vitro selection experiments and DNA modification studies using DMS and DEPC. It is preferred that the antisense molecule binds the target molecule with a dissociation constant (k _d ) of less than 10 ⁻⁶ , more preferably less than 10 ⁻⁸ , even more preferably less than 10 ⁻¹⁰ , more preferably less than 10 ⁻¹² . Representative examples of methods and techniques that aid in the design and use of various antisense molecules can be found in the following non-limiting list of US patents. (Antisense :) 5,135,917, 5,294,533, 5,627,158, 5,641,754, 5,691,317, 5,780,607, 5,786,138, 5,849,903, 5,856,103, 5,919,772, 5,955,590, 5,990,088, 5,994,320, 5,998,602, 6,005,095, 6,007,995, 6,013,522, 6,017,898, 6,018,042, 6,025,198, 6,033,910, 6,040,46,46,0,46 And 6,057,437; (Aptamer) 5,476,766, 5,503,978, 5,631,146, 5,731,424, 5,780,228, 5,792,613, 5,795,721, 5,846,713, 5,858,660, 5,861,254, No. 5,869,641, No. 5,958,691, No. 6,001,988, No. 6,011,020, No. 6,013,443, No. 6,020,130, No. 6,028,186, No. 6,030,776, No. 6,051,698, (Ribozyme :) No. 5,646,042, No. 5,535, No. 5,69 5,731,295, 5,811,300, 5,837,855, 5,869,253, 5,8 77,021, 5,877,022, 5,972,699, 5,972,704, 5,989,906 and 6,017,756. In addition, a description of suitable techniques, particularly with respect to siRNA, can be found in the following literature: U.S. Patent No. 6,506,559, International Publication WO 01/29058, International Publication WO 01/68836, International Publication WO 01/75164, U.S. Publication 20020114784, U.S. Publication 20030125281, U.S. Publication 2002162126, U.S. Publication 20030108923, US Publication No. 20020173478, Fire, et al. Nature 391: 806-811 (1998); Yang, et al., Mol. Cell. Biol. 21: 7807-7816 (2001), Elbashir, et al., Nature 411: 494-498 (2001), Hammond et al. Nat. Rev. Genet 2: 110-119 (2001), and Sharp, Genes Dev. 15: 485-490 (2001)).

          In certain aspects, the Ln-5 modulator is a target gene expression product (eg, any of cancer progression and / or development of any of the IBD, ADPKD, or other diseases or disorders described herein). SiNRA (typically a dsRNA molecule of about 20-25 nucleotides) that interferes with the action of the gene expression product associated with the polypeptide that induces modifications in Ln-5 associated with this aspect. The following DNA sequence corresponds to a typical siRNA sequence targeting the Ln-5 γ2 chain.

AGTGCTCGATGTGACAACTCT (SEQ ID NO: 1)
AGCCAGATGCGACCGAGTCT (SEQ ID NO: 2)
AGGTTCCATAAGATCACCCT (SEQ ID NO: 3)
AGCCTGGCAGAAAGTGAAGCT (SEQ ID NO: 4)
AGGAGATTGGGAGTCTGAACT (SEQ ID NO: 5)
The following correspond to random siRNA sequences that can be used as controls.

AGGGCCGACATAGATGAGACT (SEQ ID NO: 6)
AGCGTGCGCTGAAAGTGAAGCT (SEQ ID NO: 7)
how to use
As mentioned above, Ln-5 modulators can induce, promote and / or enhance physiological responses associated with changes in Ln-5 activity. These changes in physiological response or activity are then accompanied by the promotion and / or enhancement of therapeutic or prophylactic protection against various Ln-5 related diseases and disorders.

          Unless otherwise stated or clearly contradicted by context, the term “treatment” is intended to prevent the occurrence of any symptom or condition or to alleviate such a symptom or condition that has already occurred. Represents delivering an effective amount of a therapeutically active compound of the present invention for the purpose of reducing, eradicating or eradicating (curing). “Treatment” induces, promotes and / or enhances one or more physiological responses or characteristics associated with the treatment of a disease state, or the severity, transmission, or state of a disease state as described herein. It can also mean reducing the risk of onset or onset. Thus, the term “therapy” is intended to include prophylactic treatment. Unless otherwise stated or clearly contradicted, terms similar to “treatment” (eg, “treat”) should be interpreted similarly. However, the “therapeutic therapy” for the already established diseases provided by the teachings herein and the prevention of the disease, to reduce the severity of the later-onset disease, until the onset of the disease symptoms It will be apparent that such “prophylactic therapy” used to increase the time of, or any combination of, etc. is considered a separate and independent aspect of the invention (eg, Such therapies can vary in terms of dosage, dosing schedule, etc.). By way of example, prophylactic treatment, i.e., treatment aimed at reducing the risk of a patient developing a disease state, may determine the incidence of a disease state in a group to be treated during a given study period. It can be evaluated by comparing with the incidence of disease states in the group.

          As defined herein, an “effective amount” refers to treatment of a disease, condition or disease when administered to a subject suffering from a disease, condition or disease and at risk for a disease, condition or disease. Or an amount of an active agent such as an Ln-5 modulator that provides one or more physiological responses with alleviation or reduction of one or more disease symptoms in a subject. A “therapeutically effective amount” is an active agent that, when administered to a subject suffering from a disease, condition or disease, results in one or more physiological responses with treatment of symptoms of the characteristics of the established disease Is the amount. A “prophylactically effective amount” is an amount of an active factor that, when administered to a subject, suppresses or reduces the risk of developing one or more disease symptoms in the subject. As used herein, a “physiologically effective amount” is an amount of an active agent that induces, promotes and / or enhances one or more physiological responses. In embodiments relating to combination therapy using an Ln-5 modulator and a second agent, an “effective amount” of said factor, separately or in combination, is affected by a disease, condition, or disease, and a disease Each, when administered to a subject at risk for a medical condition or disease, provides one or more physiological responses with treatment of the disease, medical condition or disease, or with alleviation or reduction of one or more disease symptoms in the subject Represents the amount of the factor. In certain embodiments, the effective amount has a combined or increased potency, or is synergistically effective.

          In one aspect, the present invention provides a method of promoting prevention and / or treatment of inflammatory bowel disease (“IBD”). Thus, for example, the present invention provides Ln-5 modulators (eg, Ln-5 associated antibodies, Ln-5 binding proteins) to patients identified as having or at risk of developing significant IBD. Or a combination of both) in an amount sufficient to detectably induce, promote and / or enhance one or more physiological responses. Methods are provided for inducing, promoting and / or enhancing one or more associated physiological responses.

          Such methods of the invention can be used to induce, facilitate, promote and / or enhance the treatment and / or prevention of any aspect of IBD. Thus, in one aspect, such methods of the invention are useful in the treatment and / or prevention of ulcerative colitis (UC). Such a method of the invention can additionally or alternatively be used for the treatment and / or prevention of Crohn's disease.

          In a more specific aspect, performance of such a method of the present invention can detect a reduction (in terms of transmission and / or severity) of swelling, scarring, inflammation or any combination thereof in gastrointestinal tissue such as colon tissue. To bring. Implementation of such a method can additionally or alternatively be accompanied by a reduction in the severity and / or transmission of, for example, colitis, ileitis and / or proctitis. In another aspect, performance of such methods can reduce the size and / or number of ulcers in the patient's gastrointestinal system, such as the intima of the colon of a patient suffering from ulcerative colitis. Performing such a method can also involve reducing the frequency of bowel movement (relieving diarrhea) and / or reducing the amount of blood in the stool of the patient / host. Such a method can also involve a reduction in abdominal pain. Furthermore, implementation of such methods may also reduce the risk of developing colorectal cancer or reduce the severity of colorectal cancer. Implementation of such a method of the present invention can additionally or alternatively involve, for example, reduction of rectal bleeding and reduction of the risk of colon rupture. Methods for assessing such effects are known in the art and are discussed elsewhere herein (eg, colon barium enema and / or surgical examination). An improvement in any of the previously described disease symptoms or characteristics may be a physiological response according to the present invention.

          As mentioned above, another aspect of the present invention is to induce, enhance, and / or promote the treatment and / or prevention of IBD as LN-5 associated antibody, Ln-5 binding antibody or such It is the discovery that it can be achieved by administration of therapeutically and / or prophylactically effective amounts of Ln-5 modulators such as peptide combinations. The present invention provides Ln-5 modulators (eg, Ln-5 regulated siRNA, Ln-5 regulated antisense molecule, Ln-5) to patients identified as having or at risk of developing IBD. A composition comprising an associated antibody, Ln-5 binding protein, or any combination thereof) detectably reduces the severity, transmission, initiation, or risk of developing IBD in a population of similar patients, and / or Or reduce the risk of severe, spread, initiation or onset of IBD and / or facilitate its treatment, including administration in an amount sufficient to detectably facilitate treatment of IBD in a population of similar patients As well as methods for reducing or reducing the risk of developing complications associated with IBD, such as colorectal cancer. Prophylactic or therapeutic amounts of such compositions can be determined, for example, in connection with clinical trials for IBD. Such studies and studies of IBD patients can be used to assess the risk of developing IBD and / or the risk of mean onset time in similar patients. Appropriate clinical indicators for IBD in this regard are well known and include, for example, the degree of inflammation (eg, determined by visualization through colonoscopy or sigmoidoscopy), anemia, white blood cell count, and ulcers. Degree (for example, determined by barium enema x-ray). Suitable indicators for CD include the number and / or size of the fistula, pain around the anus, thickening of the intestinal wall, and / or “discontinuous lesion” (or simply “skips” —inflammation occurs The number and size of alternating regions). Appropriate indicators for ulcerative colitis include inflammatory polyps, granular mucosal roughness, “lead tubular colon” (colon lumen stiffness and symmetric stenosis), “burnt colon” (colon bulge pattern) -Free generalized colonic loss-associated with general loss of colonic swelling), filamentous polyposis, occult or overt blood loss, weight loss, neutrophil infiltration, loss of epithelial cells (multiple ulcers, fibers , Dysplasia and longitudinal contraction of the colon)), folate deficiency and toxic megacolon may be included. Other imaging methods (not endoscopic and / or radiological procedures) can be used to evaluate IBD. Magnetic resonance spectroscopy (MRS) is a variation of magnetic resonance imaging (MRI) that may prove useful for evaluating IBD. Computed tomography (CT) imaging and ultrasound are useful in determining the extent of disease on the intestine and may be useful in detecting advanced IBD abscesses and other complications. Any reduction of the above-described indicators can be used as a physiological response indicator in the present invention.

          The present invention improves a therapeutically effective amount of a composition comprising an Ln-5 modulator, such as an Ln-5-associated antibody, an Ln-5 binding protein, or a combination thereof, so that the quality of life of the patient can be detected. Also provided is a method for detectably improving the quality of life of a patient suffering from IBD, comprising administering for a sufficient period, under conditions and in an amount. The use of quality of life assessment in IBD treatment is well identified in the art. For example, “Han et al., Scand J Gastroenterol. 1998 Sep; 33 (9): 961-6; Guyatt et al., Gastroenterology. 1989 Mar; 96 (3): 804-10; Panwala et al., J Immunol 1998 Nov 15; 161 (10): 5733-44; Irvine et al., Gastroenterology. 1994 Feb; 106 (2): 287-96; Cheung et al., J Clin Epidemiol. 2000 Mar 1; 53 (3) Irvine, Scand J Gastroenterol Suppl. 1993; 199: 36-9; Irvine et al., Am J Gastroenterol. 1996 Aug; 91 (8): 1571-8; Hjortswang et al., Scand J Gastroenterol. 2001 Jan; 36 (1): 77-85; Pallis, Dig Liver Dis. 2000 Nov; 32 (8): 682-8; Irvine, Scand J Gastroenterol Suppl. 1995; 208: 136-40; Han et al., Am Jan Gastroenterol. 2000 Jan; 95 (1): 145-51; de Boer et al., Eur J Gastroenterol Hepatol. 1995 Nov; 7 (11): 1043-50 .; Bernklev et al., Scand J Gastroenterol. 2002 Oct; 37 (10): 1164-74; van der Eijk et al., Am J Gastroenterol. 2001 Dec; 96 (12): 3329-36; Konig et al., Eur J Gastroenterol Hepatol. 2002 Nov; 14 (11 ): 1205-15; Griffiths et al., J Pediatr Gastroenterol Nutr. 1999 Apr; 28 (4): S46-52; Otley et al. J Pediatr Gastroenterol Nutr. 2002 Oct; 35 (4): 557-63; Int J Nurs Stud. 2002 Aug; (6): 583-90; Xiao et al. Curr Gastroenterol Rep. 2002 Dec; 4 (6): 490-6; and Ferry J Pediatr Gastroenterol Nutr. 1999 Apr; 28 (4): S15-8 " I want. Appropriate periods and dosing conditions can be readily determined by those skilled in the art using known principles, examples of which are discussed elsewhere herein.

          The foregoing methods for preventing and / or treating IBD include Ln-5 modulators (eg, Ln-5 associated antibodies, Ln-5 binding proteins or both, and one or more second IBD mitigating agents and / or IBDs). A second anti-IBD agent may be further administered with (or in combination with) the composition comprising: a prophylactic agent. Any suitable second anti-IBD agent can be administered to the host or patient in any suitable manner. Examples of second anti-IBD agents include GLP-2, GLP-2 analogs, Kunitz-type inhibitors, trefoil factor 2 (TFF2) peptide (monomer or dimer), trefoil factor (TFF1) peptide ( Monomer or dimer) and trefoil factor 3 (TFF3) peptide (monomer or dimer), non-steroidal anti-inflammatory drug (NSAIDS), steroid or alcohol, tissue factor agonist, CD45 phosphatase inhibitor, 1 , 2,4-triazoles, α-aminoketones, aminosalicylates such as sulfasalazine (azurfidine), corticosteroids, and cyclosporine, prednisone, 6-mercaptopurine or azathioprine (converted to 6-mercaptopurine in the liver) Immunosuppressants, various leukotrienes, PAF and cycloo Shigenaze pathway inhibitors, as well as IL-1, IL-6 or a receptor molecule specific proteins are antibodies or cloned targeted in inflammatory cascade, such as TNF-alpha. Another approach to relieving oxidative stress due to inflammatory responses is to use nitrone related therapy (NRT).

          In certain aspects, prevention and / or treatment of IBD using Ln-5 associated antibodies, Ln-5 binding proteins and / or other Ln-5 modulator compositions comprises 5-ASA, anti-TNFα agents and antibodies ( For example, it is enhanced by concomitant administration of an agent selected from Enbrel, Remicade), an inhibitor of VCAM and an inhibitor of ICAM.

          Further useful anti-IBD agents include TRA-flc (TRAFYK) inhibitors such as RDP58 (SanStat Medical Crp.), Antisense nucleic acid agent Aricaforsene (Isis Pharmaceuticals), interferon β, infliximab, rosiglitazone, anti-IL- 12 antibodies and anti-Il-6R antibodies and ICAM-1 inhibitors such as daclizumab. Other useful second agents, as well as anti-IBD methods and methods associated with diagnosis of IBD and evaluation of anti-IBD agents are described, for example, in US Pat. Nos. 6,610,674, 6,545,056, 6,479,465, 6,326,364, No. 6,265,374, No. 6,194,465, No. 6,183,951, No. 5,968,741, No. 5,368,854, No. 6,613,751, No. 6,599,509, No. 6,558,661, No. 6,551,593, No. 6,540,993, No. 6,500,418, No. 6,482,409, No. No. 6,358,939, No. 6,291,441, No. 6,218,129, No. 6,214,373, No. 6,166,044, No. 6,143,785, No. 6,046,179, No. 6,036,978, No. 6,028,095, No. 6,025,387, No. 6,013,670, No. No. 5,932,214, No. 5,928,883, No. 5,905,081, No. 5,888,969, No. 5,834,021, No. 5,776,972, No. 5,710,181, No. 5,674,754 and No. 5,599,795.

          Any of the above IBD-related methods may be associated with surgical techniques associated with the treatment and / or prevention of IBD (eg, ileostomy, ileo-anal or ileo-rectal anastomosis (especially in the case of ulcerative colitis), forbidden ileum Surgery, colorectal colectomy, resection and / or bypass surgery, bowel transplantation, colectomy, ileal anal penetration, ileal anal “pouch” (eg, J-pouch) technology, stem cell / precursor Cell and / or tissue transplantation), and / or therapeutic nutritional programs (eg, folic acid therapy, vitamin D therapy, anti-inflammatory vitamin C therapy, vitamin B12 therapy, zinc / calcium therapy, etc., or IBD-related “nutritional supplements” It is also possible to combine with “therapy”. Suitable treatments and other materials related to IBD are described, for example, in “Haanaer and Bayless, Advanced Therapeutic Inflammatory Bowel Disease (2d ed. 2000), US Pat. No. 6,486,349 and therein. In various references. Implementation of such methods can also be combined with therapeutic diets including, for example, fat reduction, lactose reduction and / or fiber reduction.

          The invention also provides methods for preventing IBD-related cancers, delaying the onset of IBD-related cancers, reducing IBD-related cancers and / or treating IBD-related cancers. Thus, for example, the present invention provides a composition comprising an Ln-5 modulator (eg, Ln-5-associated antibody, Ln-5 binding protein, or both) at a risk of developing cancer in a similar patient population. A method is provided for reducing the risk of developing cancer in a patient suffering from inflammatory bowel disease, comprising administering to the patient in an amount sufficient to detectably reduce. In a more general sense, the present invention has a risk of developing a patient suffering from inflammatory bowel disease and having a cancer associated with inflammatory bowel disease or developing a cancer associated with inflammatory bowel disease. Detect a physiological response to a composition that includes an Ln-5 modulator (such as an Ln-5 modulatory siRNA, an Ln-5-associated antibody, an Ln-5 binding protein, or any combination thereof) in a patient identified as A method is provided for inducing, promoting and / or enhancing a physiological response associated with the prevention and / or treatment of cancer in said patient comprising administering in an amount sufficient to induce it.

          Many colorectal cancers arise from adenomatous polyps, regardless of etiology. Only adenomas are clearly precancerous, and very few such lesions progress to cancer. Many polyps give no symptoms and remain undetected clinically. Fecal occult blood can be found in less than 5% of patients with such lesions. Clinically, the probability that an adenomatous polyp will become cancer depends on the overall appearance of the lesion, its histological characteristics and its size. Adenomatous polyps can be pedunculated (stalked) or sessile (flat base). Cancer more frequently progresses to a plain polyp. Histologically, the adenomatous polyp can be tubular, villi (ie, papillary) or ductal villi. Ciliated adenomas (most of which are pedunculated) become malignant more than three times as often as tubular adenomas. The possibility that a polypoid lesion in the large intestine contains invasive cancer is related to the size of the polyp and is negligible for lesions less than 1.5 cm (less than 2%) and for lesions between 1.5 and 2.5 cm in size Moderate (2-10%) and marked (10%) for lesions greater than 2.5 cm. The present invention provides a method of reducing the risk of developing cancer in such patients by administering an LN-5 modulator.

          Further, if a patient is identified as suffering from an IBD-related cancer (eg, by biopsy or other research technique or technique that reveals cancer and / or precancerous growth), an anti-cancer agent (eg, cancer Cell-targeted apoptotic agents, tumor vaccines, antisense molecules induced against tumor suppressors or oncogenes, or anti-angiogenic growth factors and signaling regulators, cell cycle regulation, and immune responses against tumor cells Can be administered concomitantly with Ln-5 associated antibodies, Ln-5 binding proteins, other Ln-5 modulators or combinations thereof. Exemplary anticancer agents are described herein. If the patient is identified as being at significant risk of developing an inflammatory bowel disease-related cancer, the method may include a cancer prophylactic agent (eg, a so-called tumor vaccine or an oncogene antisense nucleotide inhibitor), Ln -5 associated antibodies, Ln-5 binding proteins, other Ln-5 modulators or combinations can be administered to the patient. Examples of anti-cancer therapeutics and cancer preventives are known in the art, and examples of such agents are further described elsewhere herein. In certain aspects, a method of treating and / or preventing IBD-related cancer comprises administering the composition to a population of cells having a higher than normal aneuploid DNA distribution. In another aspect, a method for treating and / or preventing IBD-related cancer comprises administering the composition to a human having increased Ln-5 expression relative to a healthy individual.

          If the patient is identified as having an IBD-related cancer, the patient is associated with administration of Ln-5 associated antibodies, Ln-5 binding proteins, other Ln-5 modulators, or combinations thereof. Surgery can be performed. Surgery can be either primary management to remove the tumor, secondary tumor reduction surgery, or symptom relief secondary surgery. Administration of Ln-5 associated antibodies, Ln-5 binding proteins, other Ln-5 modulators or combinations thereof can be performed before surgery, during surgery, after surgery, or any suitable combination thereof. is there.

          In another aspect, the present invention provides Ln-5 modulators (Ln-5-association) to patients identified as having renal cysts or patients identified as having a substantial risk of developing such cysts. A composition comprising an antibody, Ln-5 binding protein, or both) detectably reduce the risk of developing a renal cyst in a population of similar patients, detectably suppress or propagate the development of a renal cyst, Administering to the patient in an amount sufficient to detectably reduce proliferation or propagation and proliferation in said patient, reducing the risk of developing a renal cyst, inhibiting the onset, or transmitting, proliferating or A method for reducing propagation and proliferation is provided. Methods for identifying patients with such cysts or at risk for developing such cysts are known in the art (eg, McHugh et al., Radiology. 1991 Feb; 178 (2): 383 -5; King et al., Kidney Int. 2003 Dec; 64 (6): 2214-21; and Sutters et al., J Lab Clin Med. 2003 Feb; 141 (2): 91-101). For example, genetic tests have been developed and are commercially available to detect mutations in the PKD1 and PKD2 genes. This test can detect the presence of an autosomal dominant PKD mutation before the cyst develops. The most common symptoms are back and flank pain (between ribs and buttocks) and headache. Blunt pain can be temporary or permanent and can be mild or severe. Hypertension occurs early in the disease, often before cysts appear. In addition, diverticulosis (small sac over the large intestine), kidney stones and liver and pancreatic cysts, hematuria (blood in urine), urinary tract infections can occur. Reduction of any of these symptoms and / or reduction of renal cyst size or number can be a physiological response in the present invention.

          Animal models of diseases with such cysts, such as ADKPD, are known. See, for example, “Thomson et al., Am J Physiol Renal Physiol. 2003 Nov; 285 (5): F870-80 and Gattone et al. Nature Med. 2003 vol 9 no 10 pages 1323-1326”.

          In yet another aspect, the invention relates to a patient identified as having a renal cyst or having a substantial risk of developing a renal cyst with Ln-5-associated antibody, Ln-5 binding protein, other Ln-5 modulators or any combination thereof (the terms combination and any combination thereof may be used interchangeably throughout this specification unless otherwise indicated or in context) .) Inducing and promoting a physiological response in said patient comprising administering a composition in an amount sufficient to detectably induce, promote and / or enhance a physiological response. And / or a method of enhancing.

          In a further aspect, the present invention provides Ln-5 modulators (eg, Ln-5-associated antibodies, Ln-5 binding proteins, other Ln-5 modulators or any combination thereof) to patients with renal cysts. A method of reducing a pathology associated with proliferation and / or propagation of renal cysts in the patient, comprising administering to the patient a composition comprising a sufficient amount to detectably reduce the pathology To do. In this regard, the present invention provides a method for reducing such cyst migration and / or infiltration in a patient.

          In methods relating to modulating Ln-5 function in relation to renal cysts, the effectiveness of Ln-5 associated antibodies and / or Ln-5 binding proteins can be determined by the use of anti-cystic kidney agents / anti-autosomal dominant polycystic kidneys ( ADPKD) and / or concomitant administration (pre-administration, concomitant with and / or post-administration) of agents associated with the management of ADPKD. Examples of such agents include ACE inhibitors, calcium channel blockers, trimethoprim, sulfamethoxazole, chloramphenicol and ciprofloxacin, among others. Cystic penetrating antibiotics such as trimethoprim-sulfamethoxazole, ciprofloxacin and chloramphenicol may also be useful for the treatment of ADPKD and related diseases / disorders. Chronic pain from cysts can also be managed with cyst puncture and sclerosis using ethanol.

          The effectiveness of ADPKD treatment and / or prevention techniques can be assessed by ultrasonography, venous urography and renal tomography, CT scans and MRI, and other techniques discussed herein. Is possible.

          Physiological conditions associated with ADPKD that can be modulated by administration of Ln-5 modulator (eg, Ln-5 associated antibodies and / or Ln-5 binding proteins) compositions include kidney size from an expanded state Reduction, cyst number and / or size reduction, hemorrhagic cyst number reduction, flank pain reduction, urine pH increase and ureteral obstruction reduction.

          Furthermore, the present invention provides Ln − to a host suffering from IBS or identified as being at substantial risk of developing irritable bowel syndrome (“IBS”) to achieve the desired result. Reducing the risk of developing irritable bowel disease in said patient, comprising administering a therapeutically or prophylactically effective amount of 5-associated antibody, Ln-5 binding protein, other Ln-5 modulators or combinations thereof And methods of delaying the onset of irritable bowel disease in the patient, reducing its severity, preventing and / or treating inflammatory bowel disease. Patients with IBS basically belong to two broad clinical groups. Most commonly, patients have abdominal pain with changes in bowel habits consisting of constipation, diarrhea, or both. In the second group, the patient has painless diarrhea. Abdominal pain in IBS varies greatly in intensity and position, with 25% of patients located in the lower abdomen, 20% on the right, 20% on the left, and 10% on the upper abdomen. Abdominal pain in IBS is often transient and convulsive, but can overlap the background of persistent pain. Pain may be negligible enough to be negligible, or it may interfere with daily activities. However, patients with severe IBS often wake up repeatedly at night. In addition, 25-50% of patients with IBS complain of dyspepsia, heartburn, nausea and vomiting. Improvement of any of these symptoms after administration of the Ln-5 modulator of the present invention is a physiological effect.

          In yet another aspect, the invention provides a composition comprising an amount of Ln-5-associated antibody, Ln-5 binding protein, or both or other Ln-5 modulators or combinations of Ln-5 modulators, One or more physiology associated with the prevention and / or treatment of irritable bowel syndrome, comprising administering to the patient in an amount sufficient to detectably promote, enhance and / or induce a physiological effect. A method of promoting, enhancing and / or inducing sexual effects in the patient. In another sense, the present invention provides for the management of IBS (IBD and / or ADKPD) comprising periodically administering a composition comprising one or more Ln-5 modulators to achieve a desired therapeutic effect. The same can be said of the above).

          Assessment of treatment for IBS can be performed on any suitable criteria using any suitable criteria (eg, quality of life). Suitable factors for assessing IBS management / treatment are known, eg, “Graef, AWHONN Lifelines. 2003 Aug-Sep; 7 (4): 324-30; Olden et al., Rev Gastroenterol Disord. 2003; 3 Suppl 3: S3-S11; Corazziari et al., Aliment Pharmacol Ther. 2003 Sep 15; 18 (6): 569-80; Gordon et al., Dig Dis Sci. 2003 Jul; 48 (7): 1317-23 Somers et al., Gastroenterol Clin North Am. 2003 Jun; 32 (2): 507-29; Lacy et al., Arch Intern Med. 2003 Jun 9; 163 (11): 1374-5; van Zanten, Rev Gastroenterol Disord. 2003; 3 Suppl 2: S12-7; and Ron, Isr Med Assoc J. 2003 Mar; 5 (3): 201-2 ”.

The compositions of the present invention may be useful in promoting the treatment and / or prevention of other gastrointestinal diseases such as gastrointestinal endometriosis. The composition of the present invention is described in "Cash et al., Curr Gastroenterol Rep. 2003 Dec; 5 (6): 468-75; Huerta et al. Pharmacoepidemiol Drug Saf. 2003 Oct-Nov; 12 (7): 583-8. Lacy et al., Rev Gastroenterol Disord. 2003; 3 Suppl 3: S32-42 .; Gunn et al., Postgrad Med J. 2003 Mar; 79 (929): 154-8 ”, It can be administered concomitantly with the application of IBS prevention, management and / or treatment techniques. Related methods of treating, preventing, assessing and / or identifying IBS include, for example, 6,653,339, 6,638,928, 6,596,759, 6,566,369, 6,562,629, 6,429,209, 6,395,705, No. 6,369,079, No. 6,228,040, No. 6,203,797, No. 5,965,557, No. 5,900,233, No. 5,434,174, No. 4,745,131 and No. 4,239,768.
Another feature of the invention includes a combination of an Ln-5 associated antibody, an Ln-5 binding protein, other Ln-5 modulators or combinations thereof and at least one second anticancer agent or cancer preventive agent, The treatment and / or prevention of cancer can be achieved by the administration of a composition with synergistic effects, additive effects or significantly superior effects or significantly reduced side effects compared to the case where such drugs are similarly administered individually. Including methods of inducing, promoting and / or enhancing.

          In one exemplary aspect, the present invention relates to Ln-5-associated antibody, Ln-5 binding protein or both or other Ln-5 modulators or combinations of Ln-5 modulators and one or more anticancer agents. Suffering from cancer, comprising administering to the host a composition comprising a synergistically effective combination in an amount sufficient to detectably reduce the progression and / or time to recurrence of said cancer to the patient Methods are provided for reducing cancer progression in a mammalian host.

          In another exemplary aspect, the present invention relates to Ln-5 modulators (eg, Ln-5 associated antibodies, Ln-5 binding proteins, etc.) to increase the ratio of quiescent cells to invasive cells in a mammalian host. Or both) and a therapeutically effective amount of a composition comprising a synergistically effective combination of one or more anti-cancer agents, said quiescence against invasive neoplastic cells in a mammalian host comprising administering to said mammalian host A method for increasing the proportion of cells is provided. For such detection, it is possible to use tumor biopsy material or other tumor tissue, in which case BrdU or Ki67 can be used as a proliferation marker, and laminin-5 expression or matrix metalloproteinase secretion can be used. It can be used as an invasive marker.

          In a further aspect, the present invention detectably reduces the risk of developing a cancer-related tubular network, detectably extends the occurrence of a cancer-related tubular network, and / or formed in a mammalian host. In order to detectably reduce the number of expected tubular networks associated with cancer, Ln-5 modulators (eg, Ln-5-associated antibodies, Ln-5 binding proteins or both) and one or more anticancer agents A method of inhibiting the formation of a tubular network associated with cancer in a mammalian host is provided comprising administering to the mammalian host a therapeutically effective amount of a composition comprising a synergistically effective combination with.

          In yet another aspect, the present invention provides one or more with Ln-5 modulators (eg, Ln-5-associated antibodies, Ln-5 binding proteins or both) to detectably reduce the invasive potential of cancer cells. Provided is a method of reducing the invasive ability of cancer cells in a mammalian host, comprising administering to the mammalian host a therapeutically effective amount of a composition comprising a synergistically effective combination with a plurality of anticancer agents.

          In yet another aspect, the invention provides Ln-5-associated antibodies, Ln-5 binding proteins, other Ln-5 modulators or combinations thereof and one or more anti-cancer agents to achieve the desired result. Reducing cell migration, reducing tumor growth, neoplasia or pre-neoplasm in a mammalian host comprising administering to the mammalian host a therapeutically effective amount of a composition comprising a synergistically effective combination of Methods are provided for reducing cell division or reducing any combination thereof.

          In a further aspect, the invention administers a composition comprising a synergistic combination of a Ln-5-associated antibody, Ln-5 binding protein, other Ln-5 modulator, or a combination thereof and a second anticancer agent. A method of promoting remission of cancer is provided.

          In yet another aspect, the present invention provides Ln-5-associated antibodies, Ln-5 binding proteins, other to detectably modulate MAP kinase activity in a neoplastic or preneoplastic cell of a mammalian host. A neoplastic cell of a mammalian host comprising contacting said cell with a physiologically effective amount of a composition comprising a synergistically effective combination of an Ln-5 modulator or a combination thereof and one or more anticancer agents, or Methods of modulating MAP kinase activity in preneoplastic cells are provided.

          In a further aspect, the present invention relates to Ln-5-associated antibodies, Ln-5 binding proteins, other Ln-5 modulators or combinations thereof and one or more anti-cancers to achieve the desired physiological effect. Reducing the risk of developing cancer in a mammalian host, comprising administering to the mammalian host a prophylactically effective amount of a composition comprising a synergistically effective combination with a prophylactic agent, until the onset of cancerous symptoms Methods are provided that reduce time, reduce the severity of cancer diagnosed at an early stage, and / or reduce the affected area of the cancer at the onset of cancer.

          In the present invention, a cancer cell is defined by exceeding the “hay flick limit” of normal cell growth (for example, as described in “Hayflick, Exp. Cell Res., 37, 614 (1965)”). ) Any cell that abnormally divides and regenerates while growing unregulated. As used herein, “cancer progression” refers to any phenomenon or combination of phenomena that promotes or is an indicator of the transition of normal non-neoplastic cells to cancerous neoplastic cells. Examples of such phenomena include changes in the phenotype of cells that involve conversion to a pre-neoplastic phenotype where normal non-neoplastic cells are recognized and the conversion of pre-neoplastic cells to neoplastic cells. Changes in the phenotype of the cells shown are included.

          Aspects of cancer progression (also referred to herein as “stages of cancer progression”) include cell crisis, immortalization and / or failure of normal apoptosis, immortalized cells and / or Or pre-neoplastic cell growth, transformation (i.e. immortalized cells can exhibit anchorage-independent, serum-independent and / or growth factor-independent or contact inhibition-independent growth) Change or shape change indicative of cancer, aneuploidy and focus formation), proliferation of transformed cells, development of metastatic potential, migration and metastasis (eg, dissociation of cells from one place and to another site) Rearrangement), formation of new colonies, tumor formation, tumor growth, new tumor formation (formation of a new tumor that is not in contact with the source of transformed cells at an identifiable location) and any combination thereof Contains It is.

          The methods of the invention can be used to reduce, treat, prevent, or otherwise alleviate any appropriate aspect of cancer progression. The methods of the present invention are particularly useful in reducing and / or reducing tumor growth and metastatic potential, as further described herein. Methods that reduce, inhibit, or otherwise alleviate such aspects of cancer progression are preferred. A particularly preferred aspect of the present invention is the reduction of metastatic ability and / or invasive (invasive) ability of cancer cells. Another preferred aspect is the effectiveness of such methods in the treatment of cancers characterized by micrometastases.

          Detection of cancer progression can be accomplished by any suitable technique, some examples of which are known in the art. Examples of suitable techniques include PCR and RT-PCR (eg, cancer cell associated genes or “markers”), biopsy, electron microscopy, positron emission tomography (PET), computed tomography, immunoscintigraphy And other scintigraphic techniques, nuclear magnetic resonance imaging (MRI), karyotyping and other chromosomal analysis, immunoassay / immunocytochemical detection techniques (eg, discriminating antibody recognition), (eg, for cell membrane changes) Includes histology and / or histopathological assays, cell kinetic studies and cell cycle analysis, ultrasound or other acoustic detection techniques, radiation detection techniques, flow cytometry, endoscopic visualization techniques and physical examination techniques. Examples of these and other suitable techniques are described, for example, in “Rieber et al., Cancer Res., 36 (10), 3568-73 (1976), Brinkley et al., Tex. Rep. Biol. Med., 37 , 26-44 (1978), Baky et al., Anal. Quant. Cytol., 2 (3), 175-85 (1980), Laurence et al., Cancer Metastasis Rev., 2 (4), 351-74 (1983), Cooke et al., Gut, 25 (7), 748-55 (1984), Kim et al, Yonsei Med. J., 26 (2), 167-74 (1985), Glaves, Prog. Clin Biol. Res., 212,151-67 (1986), McCoy et al., Itnmunol. Ser., 53,171-87 (1990), Jacobsson et al., Med. Oncol. Tumor. Pharmacother., 8 (4), 253 -60 (1991), Swierenga et al., IARC Sci. Publ., 165-93 (1992), Hirnle, Lymphology, 27 (3), 111-3 (1994), Laferte et al., J. Cell Biochem. , 57 (1), 101-19 (1995), Machiels et al., Eur. J. Cell Biochem., 66 (3), 282-92 (1995), Chaiwun et al., Pathology (Phila), 4 ( 1), 155-68 (1996), Jacobson et al, Ann. Oncol., 6 (Suppl. 3), S3-8 (1996), Meijer et al., Eur. J. Cancer, 31A (7-8) , 1210-11 (1995), Greenman et al., J. Clin. Endocrinol. Metab., 81 (4), 1628-33 (1996), Ogunbiyi et al., Ann. Surg. Oncol., 4 (8), 613-20 (1997), Merritt et al., Arch. Otolaryngol. Head Neck Surg., 123 (2), 149-52 (1997), Bobardieri et al., QJ Nucl. Med., 42 (1), 54-65 (1998), Giordano et al., J. Cell Biochem, 70 (1), 1-7 (1998), Siziopikou et al., Breast J., 5 (4), 221-29 (1999), Rasper, Surgery, 126 (5), 827-8 (1999), von Knebel et al., Cancer Metastasis Rev., 18 (1 ), 43-64 (1999), Britton et al., Recent Results Cancer Res., 157,3-11 (2000), Caraway et al., Cancer, 90 (2), 126-32 (2000), Castillo et al., Am. J. Neuroadiol., 21 (5), 948-53 (2000), Chin et al., Mayo Clin. Proc., 75 (8), 796-801 (2000), Kau et al., J. Ortohinolaryngol. Relat. Spe., 62 (4), 199-203 (2000), Krag, Cancer J. Sci. Am., 6 (Suppl. 2), S121-24 (2000), Pantel et al., Curr. Opin. Oncol., 12 (1), 95-101 (2000), Cook et al., QJ Nucl. Med., 45 (1), 47-52 (2001), Gambhir et al., Clin. Nucl Med., 26 (10), 883-4 (2001), MacManus et al., Int. J. Radiat. Oncol. Biol. Phys., 50 (2), 287-93 (2 001), Olilla et al., Cancer Control., 8 (5), 407-14 (2001), Taback et al., Recent Results Cancer Res., 158, 78-92 (2001) '' and references therein. Are described in the references. The related technologies are U.S. Pat.Nos. 5,989,815, 5,939,258, 5,882,627, 5,829,437, 5,677,125, and 5,455,159 and international patent applications WO 01/69199, WO 01/64110, WO 01/60237, WO 01 / 53835, WO 01/48477, WO 01/04353, WO 98/12564, WO 97/32009, WO 97/09925 and WO 96/15456.

          Reduction of cancer progression may include (1) the rate of transformation of normal cells into neoplastic cells (or any aspect thereof), (2) the rate of preneoplastic or neoplastic cell growth, (3) preneoplastic and Number of cells exhibiting a neoplastic phenotype, (4) physical of cell culture medium (eg cell culture medium, tissue or organ containing pre-neoplastic and / or neoplastic cells (eg organ in a mammalian host)) Area, (5) probability that a normal cell will transform into a neoplastic cell, (6) probability that a cancer cell will progress to the next aspect of cancer progression (eg, reduced metastatic potential) or (7) any combination thereof Such a change can be detected using any of the techniques described above or appropriate similar techniques known in the art, which are Applied at an appropriate time prior to administration of the composition of the invention. Times and conditions for assaying whether reduction occurs may include several types, including types of cancer, types and amounts of Ln-5 associated antibodies and / or Ln-5 binding proteins and / or other Ln-5 modulators. Those skilled in the art will be able to apply techniques and principles known in the art and / or routine experimental procedures to appropriately determine the time and conditions for performing such assays. It will be possible to decide.

          The methods of the present invention can be used to reduce the progression of any suitable type of cancer. The method of the present invention can be used to treat cancer in prostate cancer cells, malignant melanoma cells (eg, cutaneous malignant melanoma cells, ocular malignant melanoma cells and lymph node-associated malignant melanoma cells), breast cancer cells, colon cancer cells and lung cancer cells. Can be used to reduce progression. The methods of the invention can be used to reduce cancer progression in both tumor-forming and non-tumorigenic cancers (eg, non-tumorigenic hematopoietic cancers). The methods of the present invention are particularly useful in the treatment of epithelial cancer and / or colorectal cancer, breast cancer, lung cancer, vaginal cancer, cervical cancer and / or squamous cell carcinoma (eg, in the head and neck).

          Additional features of such methods include reducing the size and / or number of tubular networks associated with Ln-5 and / or inhibiting the formation of tubular networks associated with Ln-5. Another feature of the present invention is a method of preventing less invasive neoplastic cells from producing an angiogenic phenotype. Such aspects can be combined with the general features of reduced invasiveness (invasiveness) and / or metastasis of cancer by such methods. Reduction of neoplastic and / or pre-neoplastic cell migration, decreased cell division and / or decreased cell migration by administration of the compositions of the invention is a further feature.

Anticancer agents and cancer preventive agents include IL-2, IL-21, TGF-β inhibitors, Killer Inhibitory Receptor (KIR) inhibition, anti-CD20-like rituximab, anti-CD33-like gentuzumab, anti-CD52-like alemtuzumab, IL19, IL20, tumor vaccine, interferon, (anti-) TNF-α, anti-IL-2R, anti-IL-15R, telomerase inhibitor, “crosslinking agents” such as platinum agents (cisplatin, carboplatin, etc.), expression of oncogenes Included are immunomodulators such as antisense oligonucleotides or siRNA that reduce, mutated or deregulated genes and immunomodulatory (oligo) nucleotides. Non-limiting examples of additional anticancer therapeutic and / or prophylactic and therapeutic methods include fluoropyrimidine carbamates such as capecitabine; non-polyglutamate thymidate synthase inhibitors; nucleosides such as tocladecin Analogs; folic acid antagonists such as pemetrexed disodium; taxanes and taxane analogs; topoisomerase inhibitors; polyamine analogs; mTOR inhibitors (eg rapamycin esters); alkylating agents (eg oxaliplatin); lectin inhibitors; Vitamin D analogs (such as theocalcitol); carbohydrate processing inhibitors; antimetabolite folate antagonists; thymidylate synthase inhibitors; other antimetabolites (eg, raltitrexed); ribonucleases Reductase inhibitors; dioxolate nucleoside analogs; thymylate synthase inhibitors; gonadotropin releasing hormone (GRNH) peptides; human chorionic gonadotropins; chemically modified tetracyclines (eg, CMT-3) COL-3); cytosine deaminase; thymopentine; DTIC; carmustine; carboplatin; vinblastine; temozolomide; vindesine; thymosin-α; histone deacetylase inhibitor (eg, phenylbutyrate); DNA repair agent (eg, DNA repair) Enzymes and related compositions such as Dimericin® (T4 endonuclease V-containing liposomes); gastrin peptides (and related compositions such as Gastrimune®) GMK and related compounds / compositions (eg, Knutson, Curr Opin Investig Drugs. 2002 Jan; 3 (1): 159-64 and Chapman et al., Clin Cancer Res. 2000 Dec; 6 (12): 4658 Β-catenin blockers / inhibitors and / or agents that reduce the production of β-catenin in the preneoplastic or neoplastic cell nucleus (see, eg, US Pat. No. 6,677,116) ), Agents that upregulate E-cadherin expression (or E-cadherin); agents that reduce slug (associated with β-catenin) gene expression: block, inhibit or antagonize PAI-1, or others Agents that modulate the interaction of urokinase plasminogen activator (uPA) with uPA receptors; survivin; DNA demethylating agents; “cross-linking” agents such as platinum-related anticancer agents (cisplatin, carboplatin, etc.); MAPK and Ra Agents that block anti-apoptotic signaling (eg, agents that interfere with cyclin D production and / or function), such as agents that inhibit signal transduction pathways or components thereof; Cepectabine / Xeloda, cytarabine / Ara-C, Cladribine Inhibitors such as metabolic antagonists such as / Leustatin, Fludaraine / Fludara, fluorouracil / 5-FU, gemcitabine / Gemzar, mercaptopurine / 6-MP, methotrexate / MTX, thioguanine / 6-TG, Allopurinol / Zylopri; Busulfan, Cyclophosphamide, mechlorethamine, Melphalan, thiotepa, semestine, carbo Acylating agents such as platin, cisplatin, procarbazine, dacarbazine, altretamine, lomusine, carmustine and chlorambucil; Inhibitors of microtubule and / or spindle formation such as Vinorelbine, or Taxane, such as Paxlitaxel, Docetaxel, Combrestatin, Epothilone B; RRR-
α-tocopheryl succinate; anthracyclines such as Daunorubicin / Cerubidine and Doxorubisin; idarubicin; mitomycin; pricamycin; all-trans retinoic acid, retinoic acid analogues such as 13-cis retinoic acid; inhibitors of receptor tyrosine kinases; ErbB-1 / EGFR inhibitors such as Iressa and Erbitux; ErB-2 / Her2 inhibitors such as Herceptin; c-kit inhibitors such as Gleevec; VEGF receptor inhibitors such as ZD6474 and SU6668; ErbB3 ErbB4, IGF-IR, insulin receptor, PDGFRa, PDGFRβ, Flk2, Flt4, FGFR1, FGFR2, FGFR3, FGFR4, TRKA, TRKC, c Inhibitors such as met, Ron, Sea, Tie, Tie2, Eph, Ret, Ros, Alk, LTK, PTK7; cancer-related enzyme inhibitors such as metalloproteinase inhibitors such as marimastat, Neovasstat; cathepsin B; cathepsin D dehydration Modulators of elementary enzyme activity; glutathione-S-transferase and related compounds such as glutasylcysteine synthetase and lactate dehydrogenase; proteasome inhibitors (eg, Bortezomib); tyrosine kinase inhibitors; farnesyltransferase inhibitors; HSP90 Inhibitors (eg, 17-allylaminogel-anamycin) and other heat shock protein inhibitors; mycophenolate mofetil; mycophenolate; asparaginase; calcineurin inhibitor; TOR-inhibitor; ultrakine) molecules; enkephalins (see, eg, US Pat. No. 6,737,397); SU11248 (Pfizer); BAY 43-9006 (Bayer and Onyx); inhibitors of “lymphocyte homing” mechanisms such as FTY720; Tarceva; Iressa; Glivec; thalidomide as well as adhesion molecule inhibitors (such as anti-LFA). Additional antineoplastic agents that can be used in the combination compositions and combination administration methods of the present invention include, for example, U.S. Patent Nos. 6,660,309, 6,664,377, 6,677,328, 6,680,342, 6,683,059 and 6,680,306 and those described in International Patent Application WO 2003070921.

          Where appropriate, one or more of such agents may be conjugated to an Ln-5 modulating molecule in addition or alternatively. Such conjugates are another feature of the invention.

          In one aspect, the present invention provides combination compositions and combination delivery / administration protocols that include one or more biological response modifiers (BRMs) in addition to one or more Ln-5 modulators. A BRM is generally a product (such as a cytokine or antibody) produced by a cell that stimulates or restores the ability of the immune system to act against disease factors (eg, cancer cells).

          In yet another aspect, the Ln-5 modulator and / or related compound is free, such as Ln-5 interacting integrin or a γ2 peptide binding fragment thereof (examples of such agents are known in the art). Of Ln-5 γ2 and / or γ2 fragment peptides that are delivered concomitantly with the molecule.

          Combination compositions and combination delivery methods may additionally or alternatively include anti-anergy agents (eg, small molecule compounds, proteins, glycoproteins or antibodies that destroy tolerance to tumor and cancer antigens). It is.

          Further exemplary and specific types of compositions and therapies that can be used / applied with delivery of one or more Ln-5 modulators or related compositions are sequentially described below. It will be appreciated that these classifications of drugs and methods are non-limiting and are used for convenience only. Some of the previously mentioned drugs may belong to one or more of the categories described below. Also, some anticancer agents cannot be easily classified. This includes, for example, imatinib, mesylate (Gleevec® or Glivec®), which are relatively new tyrosine kinase inhibitors. Another example of a novel anticancer agent is an agent that induces tumor cell fusion, such as measles glycoprotein and related nucleic acids (see, eg, US Pat. No. 6,750,206).

          In certain aspects, the second anticancer agent is selected from one of the following agents. Melphalan (Musphalan), Busulfan (Cis-platin), Carboplatin (Carboplatin), Cyclophosphamide (Ifphoslomide), Dacarbazine (Dacarbazine) , Alkylating agents such as lomustine or temozolamide; cytarabine, azathioprine, fludarabine, fludarabine, phosphate, fludarabine, fludarabine (5-fluorouracil), gemcitabine (Hydroxyurea), cladribine (Thioguanine), methotrexine (Methotrexine), capecitabine (V) and capecitabine (V) ), Vinblastine, docetaxel or paclitaxel; antimitotic agents such as doxorubicin, amsacrine, irinotecan, irinotecan bicin), epirubicin (Epirubicin), mitomycin (Mitomycin), mitoxantrone, idarubicin (Idarubicin), teniposide, etoposide (Etoposide) or topotecan ceotoposide (topocan) Asparaginase; Estramustine phosphate, Polyestradiol phosphate, Estradiol, Anastrozole, Exestane L, Letrozole L tropole, tamoxifen, megestrol acetic acid, medroxyprogesterone acetic acid, octreotide, cyproterone acetic acid (Cyproterone acetic acid) Hormones or antihormonal agents such as Tritorelin, Leuprorelin, Buserelin, or Goserelin; and any combination of the above compounds.

          One aspect of the present invention is embodied in a composition comprising at least one Ln-5 modulator and a suitable secondary antibody (eg, a suitable secondary anti-cancer mAb). In general, such compounds can include any secondary antibody or combination of secondary antibodies that does not significantly interfere with the specificity, selectivity and / or affinity of the Ln-5 modulator. Typically, the secondary antibody component of the Ln-5 modulator combination composition has no more than about 5%, such as no more than about 10%, for Ln-5 binding to the composition's Ln-5 modulator. Generally, it shows about 20% or less of competition.

In certain aspects, the present invention provides a combination composition comprising at least one Ln-5 modulator and at least one secondary anti-cancer monoclonal antibody. A number of suitable anti-cancer mAbs are known in the art and similar suitable antibodies can be developed against cancer-related targets. Specific examples of suitable secondary anti-cancer mAbs include anti-CD20 mAb (Rituximab and HuMax-CD20), anti-Her2 mAb (eg, Trastuzumab), anti-CD52 mAb (eg, Alemtumumab and Capath® 1H), anti-CD20 mAb EGFR mAb (eg, Cetuximab, HuMax-EGFr and ABX-EGF), Zamyl, Pertuzumab, anti-A33 antibody (see US Pat. No. 6,652,853), anti-tumor fetal protein mAb (US Pat. No. 5,688,505) No.), anti-PSMA mAb (eg, US Pat. No. 6,649,163 and Milowski et al., J Clin Oncol. 2004 Jul 1; 22 (13): 2522-31. Epub 2004 Jun. 01), anti-TAG-72 antibody (see US Pat. No. 6,207,815), anti-aminophospholipid antibody (see US Pat. No. 6,406,693), anti-neurotrophic factor antibody (US Pat. No. 6,548) , 062), anti-C3b (i) antibody (see US Pat. No. 6,572,856), anti-cytokeratin (CK) mAbs, anti-MN antibody (see, eg, US Pat. No. 6,051,226), Anti-mts1 mAbs (see, eg, US Pat. No. 6,638,504), anti-PSA antibodies (eg, Donn et al., Andrologia. 1990; 22 Suppl 1: 44-55; Sinha et al., Anat Rec. 1996) Aug; 245 (4): 652-61; and Katzenwadel et al., Anticancer Res. 2000 May-Jun; 20 (3A): 1551-5); antibodies against CA125; antibodies against integrins such as integrin β1; VCAM Antibodies / inhibitors; anti-α-v / β-3 integrins anti-quininostatin mAb; anti-aspartyl (asparaginyl) β-hydroxylase (HAAH) intrabody (see, eg, US Pat. No. 6,783,658); anti-CD3 mAb (US Pat. Nos. 6,706,265 and 6,750,325) and anti-CD3 bispecific antibodies (eg, anti-CD3 / Ep-CAM, anti-CD3 / her2 and anti-CD3 / EGP-2 antibodies—see, eg, Kroesen et al., Cancer Immunolother. 1997 Nov-Dec; 45 (3-4): 203-6); as well as anti-VEGF mAbs (eg, bevacizumab). Other possibly suitable secondary mAb molecules include alemtuzumab, edrecolomab, tositumomab, ibritumomab, tiuxetane and gemtuzumab ozogamicin. In one aspect, the invention relates to one or more antibodies, typically monoclonal antibodies, targeted to angiogenic factors such as VEGF, bFGF and angiopoietin-1 and / or their receptors; As well as monoclonal antibodies against other suitable targets (generally Reisfeld et al., Int Arch Allergy Immunol. 2004 Mar; 133 (3): 295-304; Mousa et al., Curr Pharm Des. 2004; 10 (1 ): 1-9; Shibuya, Nippon Yakurigaku Zasshi. 2003 Dec; 122 (6): 498-503; Zhang et al., Mol Biotechnol. 2003 Oct; 25 (2): 185-200; Kiselev et al., Biochemistry (Mosc). 2003 May; 68 (5): 497-513; Shepherd, Lung Cancer. 2003 Aug; 41 Suppl 1: S63-72; O'Reilly, Methods Mol Biol. 2003; 223: 599-634; al., Curr Cancer Drug Targets. 2002 Jun; 2 (2): 135-56; and international patent application WO 2004/035537).

          Where appropriate, such antibodies can be conjugated to a cytotoxin, radionuclide or another anticancer agent. Also where appropriate, the immunogenic peptide target of such antibodies can be used to induce an immune response in the combination composition or combination administration method of the invention.

          Where appropriate, targets for these secondary antibodies can also be targeted by the multispecific LN-5 modulators of the present invention. Thus, for example, VH, VL and / or CDR sequences from such mAbs are also described elsewhere in this specification as bispecific and multispecific Ln-5 modulators such as L5G2D3BP It can be used in connection with Similarly, mAbs specific for the targets discussed above for bispecific mAbs, cancer antigens and other molecules discussed for other aspects can also be used in the methods of the invention. Can be incorporated into the composition of the present invention.

          Other antibodies developed against lymphomas, white blood cells, micrometastases and solid tumors may also be useful in the combination methods and / or compositions of the present invention. Antibodies that inhibit functions essential for tumor cell survival, proliferation, invasiveness and / or migration; antibodies that induce ADCC or CDC against tumor / cancer cells; antibodies that interfere with key cancer progression-related signaling events And / or antibodies that deliver a load that is toxic to preneoplastic and / or neoplastic cells may be particularly useful in such methods and compositions. Tumor cell death can also result in the release of tumor antigens that “vaccinate” the immune system, and after a secondary reaction has occurred, can stimulate the tumor / cancer cells to be targeted. Thus, in one aspect, the present invention targets specific cancer cells or tumor populations, and subsequently monitors patients for secondary responses, and such secondary responses are considered inadequate. Thus, a method of providing additional anticancer therapy is provided. Overexpressed expressed genes and tumor specific antigens can be advantageous targets for such antibodies. Tumor antigens (“vaccines”) that may be useful as immunogenic peptides in this context or per se are, for example, “Stauss et al .: Tumor antigens recognized by T cells and antibodies. Taylor and Frances (2003) and Durrant et al., Expert Opin. Emerging Drugs 8 (2): 489-500 (2003) ”.

          In a further aspect, the present invention provides a combination treatment therapy comprising such a combination of an Ln-5 modulator and one or more anti-cancer secondary mAbs. Such secondary mAb variants, derivatives of such secondary mAbs or variants of secondary mAbs and any functional fragments thereof are additionally or alternatively used in the methods of the invention. It will be apparent that it is possible and can be incorporated into the compositions of the invention.

          In another aspect, the present invention provides combination compositions and combination therapies comprising “chemotherapeutic agents” in addition to anti-cancer Ln-5 modulators. In the present invention, chemotherapeutic agents are typically at least some cell types that are selectively toxic to and / or contain cancer cells and precancerous cells. Represents a small molecule compound that exerts one or more effects on the cell cycle.

          Combinations of chemotherapeutic agents commonly used in the treatment of cancer can also be combined with one or more Ln-5 modulators in the composition or by simultaneous delivery. For example, in one aspect, the present invention provides CHOP chemotherapy (cyclophosphamide, doxorubicin, doxorubicin, so that at least an additive anti-cancer effect is obtained) Provided is a method of treating cancer comprising delivering an effective amount of one or more Ln-5 modulators to a patient with effective application of chemotherapy comprising a combination of vincristine and prednisone.

          In another aspect, the Ln-5 modulator and / or related composition comprises a chemotherapeutic agent that acts on the progression of cancer at the DNA level (non-limiting examples of conventional chemotherapeutic agents at the DNA level). Antimetabolites (6-mercaptopurine, 6-thioguanine, methotrexate, 5-fluorouracil and hydroxyurea), cytarabine, alkylating agents, procarbazine, topoisomerase inhibitors, platinum derivatives, anthracyclines and antibiotics. ) And delivered jointly.

          In another specific aspect, the Ln-5 modulator or related composition is delivered to a subject or included in a composition, together with an “RNA level” chemotherapeutic agent (or combination thereof) Non-limiting examples of chemotherapeutic agents include L-asparaginase, vinca alkaloids, taxanes, anti-cancer taxane combination compositions (such as docetaxel + prednisone) and topoisomerase inhibitors.

          In another specific aspect, the Ln-5 modulator and / or related composition is delivered in conjunction with an effective dose of decarbazine (DTIC).

          In another specific aspect, the present invention relates to one or more Ln-5 modulators and 5-fluorouracil, actinomycin D, amsacrine, arsenic trioxide, asparaginase, azadocitadine (5-azacytidine, 5AZ) , Busulfan (milleran), capecitabine, carboplatin (paraplatin), carmustine (BiCNU), chlorambucil (Leukeran), cisplatin (Platinol), cyclophosphamide (Cyroxan), cytarabine (Ara-C), dacarbazine, dactinobicin, dactinobicin (Cerubidine), docetaxel (Taxotere), doxorubicin (Adriamycin Doxil), epirubicin (Ellence), etoposide (VP) 16, Vespid), fludarabine, fluorouracil, gemcitabine, greevac (imatinib mesylate, STI 571), hydroxyurea, idarubicin, ifosfamide (Ifex), irinotecan, liposomal doxorubicin, lomustine, mechlorecamine (Mustalgen), (6MP), methotrexate, methyl CCNU, mitomycin, mitoxantrone, nitrogen mustard, nitrosourea, oxaliplatin, paclitaxel (Taxol), pentostatin, prikamycin, procarbazine, streptozocin, telcyta (Telcyta) Alone or in combination with doxil), teniposide (Vumon), thiotepa, Retinoic, vinblastine (Velban), provides vincristine (Oncovin) and vinorelbine combination compositions and combination therapies comprising an effective amount of one or more chemotherapeutic agents selected from (Navelbine).

          To further clarify the present invention, a non-limiting group of chemotherapeutic agents useful in Ln-5 modulator combination compositions and therapy is described herein.

          In one aspect, the present invention provides combination compositions and combination therapies that include one or more alkylating agents. Alkylating agents are so named because they can add alkyl groups to many electronegative groups under conditions present in cells. These agents crosslink guanine bases in the DNA double helix-stop tumor growth by directly attacking the DNA. This prevents the chain from unwinding and separating the coil. Since this is necessary for DNA replication, cells affected by such agents can no longer divide. Examples of alkylating agents include busulfan (Myleran), busulfan injection (Busulex Injection), carboplatin (Paraplatin), carmustine injection (BiCNU Injection), chlorambucil (Leukeran), cyclophosphamide injection (Cytoxan osin, N (DTIC, DTIC-Dome), ifosfamide (Ifex), lomustine (CCNU, CeeNU), mechlorethamine (Mustergen, Nitrogen mustard), melphalan (Alkeran, L-PAM), melphalan injection (Alkeran Injecto, Alkeran Injection) ), And thiotepa included.

          In another aspect, the present invention provides combination compositions and therapies comprising one or more Ln-5 modulators and one or more antimetabolites. Antimetabolites prevent cancer cells from processing nutrients and other substances required for the normal activity of cancer cells. More specifically, the antimetabolite pretends to be a purine or pyrimidine that is a building block of DNA. They prevent cell division by preventing the uptake of these substances into DNA during the “S” phase of the cell cycle. There are several different cellular targets for antimetabolites. Some general classes of antimetabolites are folic acid antagonists, purine antagonists and pyrimidine antagonists.

          Folate antagonists (also known as folate antagonists) inhibit dihydrofolate reductase (DHFR), an enzyme involved in nucleotide formation. When this enzyme is blocked, no nucleotides are formed, destroying DNA replication and cell division. Methotrexate is the main folate antagonist used as a chemotherapeutic agent.

          Purine antagonists function by inhibiting DNA synthesis in two different ways. They can inhibit the production of adenine and guanine, which are purine-containing nucleotides. If the cells do not have a sufficient amount of purines, DNA synthesis is stopped and the cells cannot divide. They can also be incorporated into DNA molecules during DNA synthesis. The presence of the inhibitor is thought to interfere with further cell division. Examples of purine antagonists include 6-mercaptopurine, dacarbazine and fludarabine.

          Pyrimidine antagonists can be combined or co-delivered with one or more Ln-5 modulators. Pyrimidine antagonists act to block the synthesis of pyrimidine-containing nucleotides. These drugs typically have a structure similar to the natural compound they substitute. By acting as a “bait”, these drugs can interfere with the production of the final nucleotide. They can exert their effects at different stages in the pathway and can directly inhibit critical enzymes. Pyrimidine antagonists can also be incorporated into the growing DNA strand and terminate this process. Examples of pyrimidine antagonists include 5-fluorouracil, albinosylcytosine, capecitabine and gemcitabine.

          In a typical aspect, the present invention relates to floxuridine (FUDR, Fluorodeoxyuridine), fludarabine phosphate (Fludara), gemcitabine hydrochloride (Gemzar), hydroxyurea (Droxia, Hydrea), mercaptopurine (6-MP , Purinethol), methotrexate (Rheumatrex, Trexall), methotrexate injection (Amethopterin, MTX Injection), thioguanine (6-TG, TG), and any combination thereof. Combination compositions and therapies are provided.

          The present invention also relates to combination compositions and therapies comprising one or more anti-neoplastic hormones. Anti-neoplastic hormones interfere with receptors for growth stimulating proteins at the cellular level. By blocking the receptor, production or release of growth factors is reduced. Examples of such agents include diethylstilbestrol injection (Stilphostrol Injection), megestrol (Megace) and mitotan (Lysodren).

          In a further aspect, the present invention provides a combination composition and a combination delivery method characterized by comprising one or more mitotic inhibitors. Mitotic inhibitors generally inhibit cell division by interfering with a protein called tubulin. Tubulin is the basic building block of fibers required to ensure that each cell continues to grow. Examples of such agents include docetaxel (Taxotere), etoposide injection (Toposar, VePesid Injection), etoposide oral (VP-16, VePesid Oral), paclitaxel (Onxol, Taxol), vinblastine (Velban) and vincristine (Velban) Vincasar) is included.

          Another slightly related class of classic chemotherapeutic agents are plant alkaloids. These alkaloids are obtained from plants and block cell division by preventing microtubule synthesis. These are essential for cell division and without them, cell division cannot occur. The main examples are vinca alkaloids such as vincristine.

          Another slightly related class of classic chemotherapeutic agents are “genotoxic” drugs. Genotoxic drugs are chemotherapeutic agents that affect nucleic acids and change their function. These drugs can bind directly to DNA or indirectly cause DNA damage by affecting enzymes involved in DNA replication and possibly apoptosis. Such genotoxic chemotherapeutic treatments include alkylating agents, intercalating agents (interfering into the space between nucleotides in the DNA double helix, thereby interfering with transcription and replication and often inducing mutations. Drugs), and enzyme inhibitors (eg, agents that inhibit important enzymes such as topoisomerase involved in DNA replication that induce DNA damage). For this reason, the genotoxic drug class of chemotherapeutic agents overlaps with other classes described elsewhere in this specification. The ultimate goal of treatment with any of these drugs (ie, the general mechanism of action associated with such drugs) includes inducing DNA damage in cancer cells. DNA damage, if severe enough, will induce cells to undergo apoptosis. Genotoxic chemotherapeutic drugs typically affect both normal and cancer cells. The selectivity of drug action is based on the sensitivity of rapidly dividing cells (such as cancer cells) to treatments that damage DNA. Examples of drugs that can be classified as genotoxic agents include busulfan, carboplatin, carmustine, chlorambucil, cisplatin, cyclophosphamide, dacarbazine, daunorubicin, doxorubicin, epirubicin, etoposide, idarubicin, ifosfamide, irinotecan, lomustine, mechlorethamine, Melphalan, mitomycin C, mitoxantrone, oxaliplatin, temozolamide and topotecan are included.

          Another aspect of the invention is embodied in combinatorial methods and compositions comprising one or more Ln-5 modulators (or their delivery) and one or more nucleic acids (or their delivery) that act as anticancer agents. .

          In certain aspects, the present invention provides combination compositions and methods in which an Ln-5 modulator is combined with or delivered with a nucleic acid comprising a sequence encoding a tumor suppressor. In one exemplary aspect, one or more Ln-5 modulators are delivered with delivery of a p53 tumor suppressor gene (eg, Roth et al., Oncology (Huntingt). 1999 Oct; 13 (10 Suppl 5): 148-54) and Nielsen et al., Cancer Gene Ther. 1998 Jan-Feb; 5 (1): 52-63). ). Additional tumor suppressor targets include BRCA1, RB1, BRCA2, DPC4 (Smad4), p21, E2F-1, FUS1 compounds (eg, INGN 401), MSH2, MLH1 and DCC. Such nucleic acids can be delivered in the form of suitable vectors, host cells, and the like. For example, one or more Ln-5 modulators are combined with a replication-deficient adenovirus encoding human recombinant wild-type p53 / SCH58500, or with a replication-deficient adenovirus encoding human recombinant wild-type p53 / SCH58500. Can be delivered.

          A further aspect of the present invention is a combination method and composition comprising one or more Ln-5 modulators and one or more nucleic acids capable of reducing one or more aspects of expression of a particular cancer-related gene. The provision of things. Such agents include antisense nucleic acids and inhibitory RNA (iRNA) molecules.

          In one exemplary aspect, the present invention provides combinatorial compositions and methods comprising one or more antisense nucleic acids targeted to oncogenes, mutated genes or deregulated genes. In another exemplary aspect, the present invention provides combination compositions and methods comprising at least one siRNA molecule targeted to a mutated gene or a deregulated gene. Another feature of the present invention is that an oncogene or other cancer progression-related gene (eg, an antisense molecule targeted to Ln-5 or an antisense molecule targeted to integrin—see, for example, US Patent Application No. 20030243993 and O'Toole et al., Exp Cell Res. 1997 Jun 15; 233 (2): 330-9 and / or MT1-MMP antisense oligonucleotides (eg, Giles et al., J Cell Sci. 2001 Aug; 114 (Pt 16): 2967-76)), other inhibitors of Ln-5 production or activity (see e.g. p300-see eg Miller et al., J Biol Chem. 2000 Mar 17; 275 (11): 8176-82). , MT1-M Antisense molecules against Ln-5 modulators such as P antisense oligonucleotides (see Giles et al., J Cell Sci. 2001 Aug; 114 (Pt 16): 2967-76)), other of Ln-5 production or activity Inhibitors (eg, p300—eg, Miller et al., J. Biol Chem. 2000 Mar 17; 275 (11): 8176-82), reduced expression of mutated and / or deregulated genes Combination compositions and methods comprising one or more antisense oligonucleotides and / or siRNAs.

          For example, Ln-5 modulators such as L5G2D3BP include anti-cancer antisense nucleic acids (eg, Genasense® (Augmerosen / G3139), LY900003 (ISIS 3521), ISIS 2503, OGX-011 (ISIS 112989), LE-AON. / LEraf-AON (c-raf antisense oligonucleotide encapsulating liposomes / ISIS-5132), MG98, and other antisense nucleic acids targeting PKCα, clusterin, IGFBP, protein kinase A, cyclin D1 or Bcl- 2- For example, Benimetteskaya et al., Clin Prostate Cancer. 2002 Jun; 1 (1): 20-30; Tortora et al., Ann NY Acad Sci. 2003 Dec; 1002: 236-43; Gleave et al., Ann NY Acad Sci. 2003 Dec; 1002: 95-104 .; Lahn et al., Ann NY Acad Sci. 2003 Dec; 1002: 263-70; K im et al., Int J Oncol. 2004 Jan; 24 (1): 5-17; Stahel et al., Lung Cancer. 2003 Aug; 41 Suppl 1: S81-8; Stephens et al., Curr Opin Mol Ther. 2003 Apr; 5 (2): 118-22; Cho-Chung, Arch Pharm Res. 2003 Mar; 26 (3): 183-91; and Chen, Methods Mol Med. 2003; 75: 621-36)) It can be combined with or administered with the anti-cancer antisense nucleic acid. In another exemplary aspect, the Ln-5 modulator is an anti-cancer inhibitory RNA molecule (eg, Lin et al., Curr Cancer Drug for a discussion on such iRNA molecules, related principles and related methods). 2001 Nov; 1 (3): 241-7, Erratum in: Curr Cancer Drug Targets. 2003 Jun; 3 (3): 237; Lima et al., Cancer Gene Ther. 2004 May; 11 (5): 309 -16; Grzmil et al., Int J Oncol. 2004 Jan; 24 (1): 97-105; Collis et al., Int J Radiat Oncol Biol Phys. 2003 Oct 1; 57 (2 Suppl): S144; Yang et al., Oncogene. 2003 Aug 28; 22 (36): 5694-701; and Zhang et al., Biochem Biophys Res Commun. 2003 Apr 18; 303 (4): 1169-78.) Or combined in an anti-cancer inhibitory RNA molecule in the composition.

          In another aspect, the invention provides that L5G2D3BP is an angiozyme (eg, Pennati et al., Oncogene. 2004 Jan 15; 23 (2): 386-94; Tong et al., Clin Lung Cancer. 2001 Feb; 2 (3): 220-6; Kijima et al., Int J Oncol. 2004 Mar; 24 (3): 559-64; Tong et al., Chin Med J (Engl). 2003 Oct; 116 ( 10): 1515-8; and Orlandi et al., Prostate. 2003 Feb 1; 54 (2): 133-43) and herzyme (in a related sense, US Pat. No. 6,617,438) Combination compositions and combination administration in combination with anti-cancer nucleozymes such as ribozymes are provided. Additional anti-cancer ribozymes are described in US patent applications 20030195164, 2003050236 and 20030105043 and US Pat. Nos. 6,482,803 and 6,489,163. See also "Poliseno et al., Current Pharmaceutical Biotechnology August 2004, vol. 5, no. 4, pp. 361-368 (8)" for a general review of RNA-based drugs.

          In yet another aspect, the Ln-5 modulator is combined or co-delivered with an immunostimulatory nucleic acid (in another aspect, a nucleic acid comprising a sequence encoding the Ln-5 modulator and at least one immunostimulatory sequence Is provided.) Numerous examples of suitable immunostimulatory nucleic acids are described in the art (eg, Krieg, Trends in Microbiol 7: 64-65 (1999); Wooldridge et al., Curr Opin Oncol. 2003 Nov; 15 (6 ): 440-5; Jahrsdorfer et al., Semin Oncol. 2003 Aug; 30 (4): 476-82; Jahrsdorfer et al., Curr Opin Investig Drugs. 2003 Jun; 4 (6): 686-90; Carpentier et al., Front Biosci. 2003 Jan 1; 8: e115-27; U.S. Patent No. 6,406,705; U.S. Patent No. 6,218,371; U.S. Patent Application No. 20040181045; and U.S. Patent Application No. 20040087538).

          In another aspect, the invention up-regulates endogenous Ln-5 production (eg, by so-called gene activation) at least one Ln-5 encoding nucleic acid (in a form with normal cell basement membrane attachment / association). One or more Ln-5 modulators together with at least one nucleic acid and / or one or more cells expressing Ln-5 at a level at least as large as that in epithelial cells associated with a normal basement membrane Methods are provided for treating an Ln-5 association condition comprising delivering an agent and / or combining one or more Ln-5 modulators (or related compositions). Other functional gene replacement methods can be used in connection with the methods of the present invention and can be reflected in the compositions of the present invention (eg, tumor suppressors such as p53). Providing a nucleic acid encoding a non-cancer related version of). Another use of gene therapy is to introduce enzymes into these cells that make the cancer cells sensitive to certain chemotherapeutic agents (eg, to make cancer cells sensitive to acyclovir, Introducing thymidine kinase into cancer cells).

          In a further aspect, the invention provides that the Ln-5 modulator is ginsenoside-Rb2, anti-MMP-1 antibody, anti-integrin antibody, anti-MMP2 antibody and inhibitor, anti-MT1-MMP antibody and inhibitor, anti-EGF-R Antibodies, anti-BMP-1 inhibitors and antibodies, and urokinase-type plasminogen activator (uPA) and / or inhibitors of plasminogen activation to plasmin (aprotinin, amiloride, EACA, tranexamic acid, anti-uPA antibody A combination composition that is combined or co-administered with a basement membrane targeting factor and / or basement membrane association factor that modulates an anti-cancer molecule (eg, a molecule that inhibits destruction of the basement membrane during cancer progression) Products and methods of administration are provided. In another aspect, the present invention provides such compositions and methods, wherein said composition or method comprises an inhibitor of thymosin β4.

          In a further aspect, the present invention provides a combination composition wherein Ln-5 modulator is combined or co-delivered with a virus or related molecule (eg, virus-like particle, viral nucleic acid, etc.) that acts as an active agent against cancer and Combination administration methods are provided. In one aspect, the present invention provides combination compositions and methods comprising one or more Ln-5 modulators (or related compositions) and at least one oncolytic virus. Examples of such viruses include oncolytic adenoviruses and herpes viruses, which may or may not be modified viruses (see, for example, Teshigahara et al. , J Surg Oncol. 2004 Jan; 85 (1): 42-7; Stiles et al., Surgery. 2003 Aug; 134 (2): 357-64; Zwiebel et al., Semin Oncol. 2001 Aug; 28 (4 ): 336-43; Varghese et al., Cancer Gene Ther. 2002 Dec; 9 (12): 967-78; and Wildner et al., Cancer Res. 1999 Jan 15; 59 (2): 410-3) .

          Various viruses, viral proteins, etc. can be used in combination compositions and combination administration methods. In general, replication deficient viruses that can replicate only one or a few rounds in vivo and that target tumor cells can be useful components of such compositions and methods, for example. Such viral factors can include or be accompanied by a nucleic acid encoding an immunostimulant such as GM-CSF and / or IL-2. Both natural oncolytic viruses and such recombinant oncolytic viruses (eg, HSV-1 virus; reovirus; replication deficient and replication sensitive adenoviruses, etc.) are useful for such methods and compositions. (Eg, Varghese et al., Cancer Gene Ther. 2002 Dec; 9 (12): 967-78; Zwiebel et al., Semin Oncol.2001 Aug; 28 (4): 336-43; Sunarmura et al., Pancreas. 2004 Apr; 28 (3): 326-9; Shah et al., J Neurooncol. 2003 Dec; 65 (3): 203-26; and Yamanaka, Int J Oncol. 2004 Apr; 24 (4): See 919-23).

          In a further aspect of the invention, the LN-5 modulator comprises a cancer antigen / tumor associated antigen (eg, epithelial cell adhesion molecule (Ep-CAM / TACSTD1), mucin 1 (MUC1), carcinoembryonic antigen (CEA), Tumor-associated glycoprotein 72 (TAG-72), gp100, Melan-A, MART-1, KDR, RCAS1, MDA7, CEA, cyclin-dependent kinase 4, β-catenin, caspase-B (capsase-B), tyrosinase, Further examples of cancer-related virus vaccines (eg, human papillomavirus vaccines), tumor-derived heat shock proteins, etc. are described elsewhere herein (eg, Acres et al., Curr Opin Mol Ther 2004 Feb, 6: 40-7; Taylor-Papadimitriou et al., Biochim Biophys Acta. 1999 Oct 8; 1455 (2-3): 301-13; Emens et al., Cancer Biol Ther. 200 3 Jul-Aug; 2 (4 Suppl 1): S161-8; and Ohshima et al., Int J Cancer. 2001 Jul 1; 93 (1): 91-6) Alternatively, combination administration methods and combination compositions to be delivered are included. Many other suitable cancer antigens / tumor associated antigens described herein (eg, gp75) and similar molecules known in the art may be used in addition to or in lieu of such combinations. It can be used in administration methods or incorporated into such combination compositions.

          Anti-cancer immunogenic peptides include BEC2 anti-idiotype mAb (Mitumomab-eg Chapman, Curr Opin Investig Drugs. 2003 Jun; 4 (6): 710-5 and McCaffery et al., Clin Cancer Res. 1996 Apr; 2 (4): 679-86), CeaVac® and related anti-idiotype mAbs (see, eg, Foon et al., J Clin Oncol. 1999 Sep; 17 (9): 2889-5), MG7 mAb Anti-idiotype mAbs against (see, eg, Fengtian et al., Chin Med Sci J. 2002 Dec; 17 (4): 215-9), and other anti-cancer anti-idiotype Abs (eg, Birebent et al., Vaccine 2003 Apr 2; 21 (15): 1601-12, Li et al., Chin Med J (Engl). 2001 Sep; 114 (9): 962-6, Schmitt et al., Hybridoma. 1994 Oct; 13 ( 5): 389-96, Maloney et al., Hybridoma. 1985 Fall; 4 (3): 191-209, Raychardhuri et al., J Immunol. 1986 Sep 1; 137 (5): 1743-9, Pohl et al ., Int J Cancer. 1992 Apr 1; 50 (6): 958-67, Bohlen et al., Cytokines Mol Ther. 1996 Dec; 2 (4): 231-8, and Maruyama, J Immunol Methods. 2002 Jun 1; 264 (1-2): 121-33) and other anti-idiotype “vaccines” are also included. Such anti-idiotype Abs are synthetic (typically inactive) molecular carriers, carriers that can be proteins (eg, keyhole limpet hemocyanin (KLH) (eg, Ochi et al., Eur J Immunol. 1987 Nov; 17 (11): 1645-8) or cells (eg erythrocytes-see eg Wi et al., J Immunol Methods. 1989 Sep 1; 122 (2): 227-34) as required. It can be advantageously conjugated.

          Compositions and combined administration methods of the present invention include the inclusion or co-administration of nucleic acid vaccines such as naked DNA vaccines encoding such cancer antigens / tumor associated antigens (eg, US Pat. No. 5,589,466). No. 5,593,972, 5,703,057, 5,879,687, 6,235,523 and 6,387,888).

          In another aspect, the combination administration method and / or combination composition comprises an autovaccine composition. In a further aspect, the combination composition and / or combination administration method comprises a complete cell vaccine or cytokine-expressing cells (eg, recombinant IL-2 expressing fibroblasts, recombinant cytokine-expressing dendritic cells, etc.) (eg, Kowalczyk et al., Acta Biochim Pol. 2003; 50 (3): 613-24; Reilly et al., Methods Mol Med. 2002; 69: 233-57; and Tirapu et al., Curr Gene Ther. 2002 Feb; 2 (1): 79-89). Another example of a therapeutic autologous cell method that may be useful in the combination methods of the present invention is the MyVax® Personalized Immunotherapy method (formerly GTOP-99) (Genitope Corporation-Redwood City). , CA, USA) (see US Pat. Nos. 5,972,334 and 5,776,746).

          In a further aspect, the combination composition and / or combination administration method of the present invention comprises administration of an immunomodulatory compound or a modulator thereof (eg, an anti-inhibitory immunomodulating antibody). Examples of such compounds include T cell activation and proliferation promoting molecules such as B7 molecules (B7-1, B7-2, variants and fragments thereof) (eg, Adv Exp Med Biol. 2000; 465: 381-90 and US Patent Application No. 2003028058), ICOS (inducible costimulator) molecules, and OX40 molecules (Coyle et al., Springer Semin Immunopathol. 2004 Feb; 25 (3-4): 349-59 and No. 6,312,700). Another example of such a molecule is a negative T, such as an antibody against CTLA4 such as MDX-010 (Phan et al. (2003) Proc. Natl. Acad. Sci. USA 100: 8372). It is an inhibitor of cell regulators. Antibodies against several members of the TNF receptor (TNFR) family have been shown to enhance T cell proliferative responses. Antibodies against CD27 have also been shown to enhance T cell proliferation. The interaction of integrin family member LFA-1 (Lymphocyte function-related antigen 1 or CD18 / CD11a) with its ligand, intercellular adhesion molecules (ICAM) -1, -2 and -3, is the activation of T cells. It is well known that it is an important factor in SLAM (signaling lymphocyte activation molecule, or CDw150) is another T cell regulator. The thermostable antigen (HSA or CD24) is a large glycosylated 38-70 kDa form that enhances T cell proliferation and is a 27 amino acid glycophosphatidylinositol (GPI) found on the surface of hematopoietic cells and neurons. It is a binding protein. 4-1BB is a costimulatory receptor for T cells. TNFR-related factor (TRAF) is also a T cell signaling molecule. CD40L can also act as an immunomodulator. The CD2-LFA3 pathway is also important for T cell control (thus factors acting on it can be included in combination methods and compositions). NK cell activation and growth factors such as stimulatory KIR molecules can also be included in such combination methods and compositions. In addition to or in place of these, other immunomodulators that can be included in such combination compositions and methods are TGFβ inhibitors.

          Cytokines and chemokines, an important subset of immunomodulators, are discussed in detail in the following discussion.

          The present invention provides combination compositions and combination delivery methods comprising at least one Ln-5 modulator and at least one anticancer cytokine, chemokine or a combination thereof.

          In general, any suitable anti-cancer cytokine and / or chemokine can be used and / or combined with the Ln-5 modulator in the methods and compositions of the invention. Appropriate chemokines and cytokines detectably result in greater and / or more comprehensive immune responses against cancer cells or related tissues (eg, tumors) in vivo, and binding of L5G2D3BP in the compositions / methods Is not substantially disturbed.

          Examples of suitable cytokines and growth factors include interferons (eg, IFNβ, IFNα (eg, INFα2b) and IFNγ (eg, IFNγ1b)) and interleukins (eg, IL-2, IL-4, IL-6, IL- 7, IL-10, IL-12, IL-13, IL-15, IL-18, IL-23, IL-24, IL-27, IL-28a, IL-28b, IL-29, etc.) . Additional cytokines that can be included in such compositions and methods include KGF, IFNβ, GM-CSF, CD40L, Flt3 ligand, stem cell factor, ancestim and TNFα (eg, Dranoff, Nat Rev Cancer. 2004 Jan; 4 (1): 11-22 and Szlosarek, Novartis Found Symp. 2004; 256: 227-37; discussion 237-40, 259-69).

          Suitable chemokines can include Glu-Leu-Arg (ELR) negative chemokines such as IP-10, MCP-3, MIG and SDF-1α obtained from the human CXC and C-C chemokine families. Suitable cytokines include cytokine derivatives, cytokine variants, cytokine fragments and cytokine fusion proteins (eg, Eliason, BioDrugs, 2001; 15 (11): 705-11 (for PEGylated cytokines) and Shibuya et al. , Laryngoscope. 2003 Nov; 113 (11): 1870-84 (other cytokine derivatives) and WO 01/79258 (albumin-cytokine fusion protein)).

          As used herein, these and other methods involving nucleic acids encoding naturally-occurring peptides are alternatively or additionally described in US Pat. Nos. 5,968,502, 6,063,630. And 6,187,305 and those described in European Patent Publication No. 0 505 500 can be performed by “gene activation” and homologous recombination gene upregulation techniques. Further useful cytokines for such combination therapies and compositions are described elsewhere herein.

          In another aspect, the invention typically provides a combination composition or combination administration comprising an Ln-5 modulator and an adjuvant, typically in combination with an anti-cancer immunogenic peptide (or surrogate nucleic acid / nucleic acid encoding molecule). Provide law. Non-limiting examples of suitable adjuvants include QS21, GM-CSF, SRL-172, histamine dihydrochloride, thymocartin, Tio-TEPA, monophosphoryl-lipid A / mycobacterial compositions, Alum, Incomplete Freund's Adjuvant, Montanide ISA, Ribi Adjuvant System, TiterMax Adjuvant, Syntex Adjuvant Formulation, Immune Stimulating Complexes (ISCOMs), GerbuR Adjuvant, CpG Oligodeoxynucleotide, Lipopolysaccharide and Polyinosine: Polycysinic Acid.

          Combination compositions and combination administration methods can also include “complete cell” and “adoptive” immunotherapy and “in vivo vaccination techniques”. For example, such methods may involve immune system cells (eg, tumor infiltrating lymphocytes (TIL), antibody expression such as CD4 + and / or CD8 + T cells (eg, T cells expanded by tumor specific antigen and / or gene modification). B cells or other antibody producing / presenting cells, dendritic cells (eg, anti-cytokine expressing recombinant dendritic cells, dendritic cells cultured with DC proliferating agents such as GM-CSF and / or Flt3-L, and / or Or dendritic cells loaded with tumor-associated antigens), anti-tumor NK cells (so-called hybrid cells) or combinations thereof (eg Fishman et al., Expert Rev Anticancer Ther. 2003 Dec; 3 (6): 837-49 Whiteside et al., Cancer Immunol Immunother. 2004 Mar; 53 (3): 240-8; Conrad et al., Curr Opin Mol Ther. 2003 Aug; 5 (4): 405-12; Trefzer et al., Mol Biotechnol. 2003 Sep; 25 (1): 63-9; Reinhard et al., Br J Cancer. 2002 May 20 ; 86 (10): 1529-33; Korbelik et al., Int J Cancer. 2001 Jul 15; 93 (2): 269-74; Costa et al., J Immunol. 2001 Aug 15; 167 (4): 2379 Hanson et al., Immunity. 2000 Aug; 13 (2): 265-76; Matsui et al., Int Immunol. 2003 Jul; 15 (7): 797-805; and Ho et al., Cancer Cell 2003 May; 3 (5): 431-7)))) or injection. Cell lysates may also be useful in such methods and compositions. Cellular “vaccines” in clinical trials that may be useful in this aspect include Canvaxin®, APC-8015 (Dendreon), HSPPC-96 (Antigenics) and Melacine® cell lysates. included. Antigens shed from cancer cells and mixtures thereof optionally mixed with adjuvants such as alum (see, eg, Bystryn et al., Clinical Cancer Research Vol. 7, 1882-1887, July 2001), such as And may be an advantageous component in the process. US Pat. No. 6,699,483 provides another example of full cell anticancer therapy. Another example of such complete cell immunotherapy that can be usefully combined in Ln-5 modulator related compositions and methods is described elsewhere herein.

          In another aspect, the one or more Ln-5 modulators are delivered to the patient with delivery of an effective amount of antigen-pulsed dendritic cells or other anti-cancer immune cells (eg, NK cells). .

          In yet another aspect, the Ln-5 modulator can be delivered to a patient in combination with application of an internal vaccination method. In vivo vaccination typically involves (i) whole tumor cells, or (ii) (a) secreted proteins, glycoproteins or other products, (b) membrane-bound proteins or glycoproteins or membranes Resulting in the induction of an immune response induced against a portion of the tumor cells, including other components associated or inserted into the membrane, and / or (c) intracellular proteins or other intracellular components, Represents induced death of tumor or cancer cells in a patient (such as cell death of tumor cells induced by drugs or induced by radiation). The immune response induced by in vivo vaccination can be humoral (ie, antibody-complement mediated) or cell mediated (eg, endogenous cytotoxicity T that recognizes tumor cells killed in the body or parts thereof). Lymphocyte development and / or increase). In addition to radiation therapy, non-limiting examples of drugs and agents that can be used to induce the induction of tumor cell death and in vivo vaccination methods include conventional chemotherapeutic agents, cell cycle inhibitors, antiangiogenic agents. Herbal medicines, monoclonal antibodies, apoptosis inducers and signal transduction inhibitors are included.

          In another aspect, the present invention provides a combination composition and combination administration method comprising at least one Ln-5 modulator and one or more cell cycle regulator / apoptosis regulator (or cell cycle / apoptosis “regulator”). provide.

          Cell cycle regulator / apoptosis regulators that can be combined with Ln-5 modulators include, for example, (i) cdc25 (non-limiting examples include NSC663284 (eg, Pu et al (2003) J Biol Chem 278, 46877). (Ii) Cyclin-dependent kinase that overstimulates the cell cycle (non-limiting examples of which are flavopiridol (L868275, HMR1275; Aventis)), 7-hydroxystaurosporine (UCN- 01, KW-2401; Kyowa Hako Kogoyo) and roscovitine (R-roscovitine, CYC202; Cyclcel) -Fischer & Gianella- Borradori (2003) Exp Op Invest Drugs 12 Reviewed in 955-970) have been described. ) And (iii) telomerase modulators (such as BIBR1532 (Dam et al (2001) EMBO J 20, 6958-6968) and SOT-095 (Tauchi et al (2003) Oncogene 22, 5338-5347), etc.) / One or more molecules that target and regulate apoptosis regulators may be included. Mycobacterial DNA has been reported to be capable of inducing apoptosis in cancer cells (see, eg, US Pat. No. 6,794,368).

          Non-limiting examples of molecules that interfere with the apoptotic pathway include TNF-related apoptosis-inducing ligand (TRAIL) / apoptotic-2 ligand (Apo-2L), an antibody that activates the TRAIL receptor , IFN and antisense Bcl-2 (Igney and Krammer (2002) Nature Rev. Cancer 2, 277-288; Makin; Dive (2003) Trends Mol Med 9, 2519; Smyth et al (2003) Immunity 18, 1-6; and Panaretakis et al. (2003) Oncogene 22, 4543-4556).

          In another aspect, the present invention provides combination compositions and combination delivery methods comprising, in addition to at least one Ln-5 modulator or related molecule, a telomerase inhibitor, a telomerase vaccine or a combination thereof. Examples of such compositions and related techniques are described in US Pat. Nos. 6,440,735 and 6,713,055.

          In yet another aspect, the present invention provides a combination composition and combination administration method comprising one or more growth factor inhibitors.

          Numerous antibodies (eg, mAbs) to growth factors and growth factor receptors are known that can be useful in promoting the treatment of cancer. For example, antibodies against the extracellular ligand binding domain of epidermal growth factor receptor (EGF-R) protein that is abnormally activated in epithelial tumors may be useful in the treatment of invasive epithelial cell derived tumors. Antibodies to low molecular weight molecules and small molecules that inhibit the tyrosine kinase domain of such receptors may also be useful in combination compositions or combination administration methods. Non-limiting examples of such molecules include Herceptin (monoclonal antibody), Cetuximab (monoclonal antibody), Tarceva (small molecule low molecular weight inhibitor) and Iressa (small molecule low molecular weight inhibitor). Additional related and useful antibodies suitable for inclusion in such combination compositions and administration methods are described elsewhere herein.

          In a further aspect, the invention relates to one or more Ln-5 modulators (or related molecular surrogates) and one or more inhibitors of angiogenesis, neovascularization and / or other angiogenesis (this Such agents are referred to herein by terms such as anti-angiogenic agents, anti-angiogenic agents, etc.)) and provide combination compositions and methods. Non-limiting examples of such agents include endostatin and angiostatin (reviewed individually or in combination) (reviewed in Marx (2003) Science 301, 452-454) and their derivatives / Anti-angiogenic heparin derivatives and related molecules (eg, heperinase III); VEGF-R kinase inhibitors and other anti-angiogenic tyrosine kinase inhibitors (eg, SU011248-Rosen et al., Clinical Oncology; May 31- June 3, 2003, Chicago, IL, USA (abstract 765)); Temozolomide; Neovasstat® (Gingras et al., Invest New Drugs. 2004 J n; 22 (1): 17-26); Angiozyme® (Weng et al., Curr Oncol Rep. 2001 Mar; 3 (2): 141-6); NK4 (Matsumoto et al., Cancer Sci. 2003 Apr; 94 (4): 321-7); macrophage migration inhibitory factor (MIF); cyclooxygenase-2 inhibitor; resveratrol (eg, Sala et al., Drugs Exp Clin Res. 2003; 29 (5-6): 263-9); PTK787 / ZK 222584 (e.g., Klem, Clin Collective Cancer. 2003 Nov; 3 (3): 147-9 and Zips et al., Anticancer Res. 003 Sep-Oct; see 23 (5A): 3869-76); anti-angiogenic soy isoflavones (see, eg, Genistein, eg, Sarkar and Li, Cancer Invest. 2003; 21 (5): 744-57); Olypraz); thalidomide and thalidomide analogs (see, eg, CC-5013—see, for example, Tohnya et al., Clin Prostate Cancer. 2004 Mar; 2 (4): 241-3); other endothelial cell inhibitors (eg, squalamine). And 2-methoxyestradiol); fumagillin and its analogs; somatostatin analogs; pentosan polysulfate; tecogalan sodium; molecules that block matrix dysfunction (suramin and analogs thereof) For example, Marchetti et al., Int J Cancer. 2003 Mar 20; 104 (2): 167-74, Meyers et al., J Surg Res. 2000 Jun 15; 91 (2): 130-4, Kruger and Figg, Clin Cancer Res. 2001 Jul; 7 (7): 1867-72, and Gradishar et al., Oncology. 2000 May; 58 (4): 324-33); Dalteparin (Scheinowitz et al. , Cardiovasc Drugs Ther. 2002 Jul; 16 (4): 303-9); matrix metalloproteinase inhibitors (such as BMS-275291-Rundhaug, Clin Cancer Res. 2003 Feb; 9 (2): 551-4; generally, Coussens et al. al. Science 2002; 295: 2387-2392); angiocol; anti-PDGF mAb and other PDGF (platelet derived growth factor) inhibitors; and PEDF (pigment epithelial derived growth factor).

          In another aspect, the invention provides that at least one Ln-5 modulator is an anti-androgen and / or anti-estrogen therapeutic agent or therapy (eg, Trachtenberg, Can J Urol. 1997 Jun; 4 (2 Supp 1): 61- 64; Ho, J Cell Biochem. 2004 Feb 15; 91 (3): 491-503), tamoxifen, progestin, luteinizing hormone-releasing hormone (or its analogs or other LHRH agonists) or aromatase inhibitors (eg, Combination compositions and combinations that are combined with or delivered with a hormone regulator such as Dreicer et al., Cancer Invest. 1992; 10 (1): 27-41) To provide a method of administration. Steroids (often dexamethasone) can inhibit tumor growth or concomitant edema (brain tumor) and may also be suitable for combination with Ln-5 modulators (or related compound surrogates) . One or more Ln-5 modulators may also be antiandrogens such as Flutamine / Eulexin; hydroxyprogesterone caproate, medroxyprogesterone / provera, megestrol acepate / Megeth Progestins such as (Megace); corticosteroids such as hydrocortisone and prednisone; luteinizing hormone-releasing hormone (LHRH) analogs such as buserelin and goserelin; and / or hormone inhibitors such as octreotide / sandstatin; Or they can be combined. In certain aspects, the Ln-5 modulator is an estrogen receptor such as tamoxifen, idoxifene, fulvestrant, droloxifene, toremifene, raloxifene, diethylstilbestrol, ethinylestradiol / estinyl, etc., or any combination thereof Provided or combined with an anticancer agent that is a modulator (ERM). Combination compositions and combination administration methods can additionally or alternatively include tamoxifen. Further teachings on cancer immunotherapy can be found, for example, in “Berczi et al.,“ Combination Immunotherapy of Cancer ”in NEUROIMMUNE BIOLOGY, Volume 1: New foundation of Biology, Berczi I, Gorczynski R, Editors, Elsevier, 2001; pp. 417- 432 ".

          In certain aspects, one or more Ln-5 modulators (or appropriate related molecular surrogates thereto-unless otherwise stated or clearly contradicted, throughout this specification, 5 such modulators are envisaged.) May be combined with one or more aromatase inhibitors such as anastrazol / arimidex, aminoglutethimide / citradene, exemestane or one or more Delivered with an aromatase inhibitor. Anti-aromatase agents inhibit the cytochrome P450 component of the aromatase enzyme complex by interfering with electron transfer from NADPH. Examples of such agents include anastrozole (Arimidex) and letrozole (Femara). These drugs can also be classified into first generation (eg, aminoglutethimide), second generation (eg, formestane and fadrazole) and third generation (anastrozole, letrozole and exemestane) compounds. . Anti-aromatase agents can also be classified as type I and type II inhibitors. Type I inhibitors have a steroidal structure similar to androgens and irreversibly inactivate the enzyme by blocking the substrate binding site and are therefore known as aromatase inactivators. Examples of such drugs include formestane and exemestane (Aromasin). Type II inhibitors are nonsteroidal and their action is reversible. Examples include anastrozole and letrozole. In one aspect, the one or more Ln-5 modulators are selected from formestane, exemestane, aminoglutethimide, anastrozole and letrozole. Provided or combined with one or more of such molecules.

         Prostate cancer is often sensitive to finasteride, an agent that blocks the peripheral conversion of testosterone to 5-hydroxy-testosterone. Ln-5 modulators can be provided with or combined with the agent, or provided with various forms of androgen depletion therapy (ADT).

          In one aspect, one or more Ln-5 modulators are combined or co-delivered with one or more intracellular signaling inhibitors. Examples of such compounds include tyrosine kinase inhibitors (Gleevec®, imatinib mesylate), modulators of the ras signaling pathway and regulators of protein transport. Other examples include serine / threonine kinase inhibitors, protein-tyrosine phosphatase inhibitors, bispecific phosphatase inhibitors and serine / threonine phosphatase inhibitors.

          In another aspect, the present invention provides combination compositions and combination delivery methods comprising one or more immune system inhibitors and one or more Ln-5 modulators. A number of immunosuppressive / immunomodulating agents are known, examples of which include T lymphocyte homing modulators (eg, FTY-720—eg, Yangawa et al., J Immunol. 1998 Jun 1; 160 (11): 5493-9); calcineurin inhibitors (such as valspodar, PSC 833 and other MDR-1 or p-glycoprotein inhibitors); and TOR-inhibitors (eg sirolimus, everolimus and Rapamycin) is included.

          Another feature of the present invention is a combination composition and combination delivery method comprising one or more Ln-5 modulators and one or more antineoplastic antibiotics. Such antibiotic chemotherapeutic agents inhibit or delay cell replication. There are a number of different antitumor antibiotics, but in general, (1) bind to DNA, making it impossible to separate DNA, (2) inhibit ribonucleic acid (RNA) and suppress enzyme synthesis It suppresses cell division in two ways. Examples of such agents include bleomycin (brenoxane), dactinomycin (actinomycin D, cosmegen), daunorubicin (servidin), doxorubicin (adriamycin, rubex), idarubicin (idamycin), mitomycin (mitomycin-C, mutamycin) ), Mitoxantrone (Novantrone), pentostatin (Nipent), plicamycin (Myrasin, Mythramycin) and combinations thereof.

          In other aspects, the Ln-5 modulator or related composition comprises low molecular weight heparin, standard heparin, pentasaccharide, thrombin inhibitors (melagatran, ximelagatran, etc.), and / or factor VII, factor VIII, etc. Delivered to the host with a thrombosis regulator such as a clotting factor.

          Because these cells may die or lack a sufficient blood supply, chemotherapeutic agents may lack the ability to kill them against a sufficiently invasive tumor. Anaerobic bacteria such as Clostridium novii can destroy the inside of tumors that are poor in oxygen. Such bacteria die when contacted with the oxygenated side of the tumor, which means they are likely to be harmless to the rest of the body. The application of such bacteria and one or more Ln-5 modulators represents another aspect of the present invention. Typically, such methods are performed in further combination with chemotherapeutic agents.

          As described above, various methods effective for the treatment of cancer can be combined with delivery of an effective amount of one or more Ln-5 modulators to a subject. Specific examples of such techniques are described in more detail herein.

          In one aspect, the invention provides a combination method comprising applying radiation to a patient or concomitantly administering a radiopharmaceutical to a patient in combination with one or more Ln-5 modulators. In a related aspect, the present invention provides a composition comprising an effective combination of one or more Ln-5 modulators and one or more radiopharmaceuticals.

          The radiation source in such a method may be either external or internal to the patient being treated (eg, radiation therapy is, for example, external beam radiation therapy (EBRT) or brachytherapy (BT). ). Radioactive elements that can be used in carrying out such methods and can be included in such compositions include, for example, radium, cesium-137, iridium-192, americium-241, gold- 198, cobalt-57, copper-67, technetium-99, iodine-123, iodine-131 and indium-111. In addition, useful radionuclides that can be incorporated into radiopharmaceuticals and can be used in such methods can be found elsewhere in the specification (eg, in connection with Ln-5 modulator conjugates). Is discussed. In certain aspects, such methods and compositions further optionally include one or more radiation protection substances. These drugs are designed to protect normal cells from radiation. An example is the intravenous drug amifostine (Ethyol). In another specific aspect, intensity-modulated radiation therapy (IMRT) is applied in combination with delivery of one or more Ln-5 modulators. IMRT delivers radiation therapy targeted to the shape of the tumor, so that damage to healthy tissue is minimized. More specifically, IMRT is capable of splitting radioactive rays and delivering them at different intensities and directions to suit the shape of the tumor. Another similar technique, three-dimensional conformal radiation therapy, has proven effective in several situations and can be combined with Ln-5 modulator therapy and prophylactic treatment regimes. In addition or alternatively, pulsed delivery of radiation (gating) can be used in this manner, reducing the field of radiation by compensating for natural breathing patterns.

          In a further aspect, the Ln-5 modulator is provided with application of radiotherapy. An example of such a therapy is of a prodrug or gene with a radiation-inducible promoter (such a gene may encode a cytotoxic protein that is an enzyme that activates the co-delivered prodrug and the like). Local production of cytotoxic factors by radiation stimulation / activation. Another example is a target Auger-emitting radiolabeled molecule. These therapies can modulate cancer by delivering targeted radiation to cells with specific receptors. Auger electrons are emitted by radioactive isotopes (iodine-125 or indium-111). Since these electrons have a very short range, they can be delivered to a specific group of target cells, leaving healthy cells. In another exemplary method, a nucleic acid comprising a radiation-inducible gene sequence encoding a protein that can be targeted by a cytotoxic factor is delivered with one or more Ln-5 modulators. Radiation is applied to produce protein and cytotoxic factors are delivered to provide targeted therapy.

          In a further aspect, the Ln-5 modulator is delivered in connection with the application of photodynamic therapy. In general, such therapies involve the delivery of photosensitizers that increase the photosensitivity of the cells, thereby causing destruction of the cancer cells when laser light is directed over the cancer area. Thus, the various prophylactic and therapeutic therapies of the present invention can be combined with anti-cancer-induced photodynamic therapy in addition or alternatively (eg, anti-cancer laser therapy—as needed) The photosensitizer can be used, for example, see Zhang et al., J Control Release. 2003 Dec 5; 93 (2): 141-50). For example, rhodium compounds are capable of damaging DNA in living cells in a manner similar to platinum classic chemotherapeutics, while being harmless until irradiated with light.

Lasers can also be used in performing accurate anti-cancer surgery (eg, in this case, labeled Ln-5 modulators identify the growth of cancerous tissue and / or precancerous tissue). . In addition or alternatively, other forms of surgery may be applied in connection with delivery of one or more Ln-5 modulators. Anti-cancer surgical techniques (eg, colectomy, colorectal resection, polypectomy, prostatectomy, segmental resection, lobectomy, lung resection, mammary tumor resection, mastectomy, etc.) are well known in the art and are therefore described herein. (E.g. CANCER SURGERY, Harvey and Beatie (WB Saunders Company 1996); ADVANCED ONCOLOGIC SURGERY, Roh et al. Eds. (Mosby-Year Books, 1st Ed. 1994); CANCER SURGERY, McKenna et al Eds. (Lippincott Williams & Wilkins 1994); The MD ANDERSON SURGICAL ONCOLOGY HANDBOOK, Feig et al. (Lippincott Williams &Wilkins; 3rd Ed. 2002); and SURGICAL ONCOLOGY: CONTEMPORARY PRINCIPLES AND PRACTICE, Bland et al. Hill Professional; 1 ^st Ed. 2001) In certain aspects, Ln-5 modulator therapy and / or labeled Ln-5 modulator diagnostic techniques are combined with anti-cancer cryosurgery. Ln-5 modulator related to anticancer therapy Then, the hormone-producing organ (such as ovary or testis) can be removed.

          In a further aspect, Ln-5 modulator therapy is combined with bone marrow transplantation and / or application of anti-cancer stem cell therapy. For example, stem cell transplantation (SCT) can be advantageously used in cancer treatment. The SCT can be autologous (previously preserved own cells), allogeneic (cells donated by another person) or syngeneic (cells donated by the same twin). SCT methods and related principles are known in the art (eg, Georges et al., Int J Hematol. 2003 Jan; 77 (1): 3-14; Tabbara et al., Anticancer Res. 2003 Nov-Dec; 23 (6D): 5055-67; Bhatia et al., Expert Opin Biol Ther. 2001 Jan; 1 (1): 3-15; Huugen et al., Neth J Med. 2002 May; 60 (4): 162- 9; Margolin et al., J Urol. 2003 Apr; 169 (4): 1229-33; and US Pat. No. 6,143,292). Bone marrow transplantation is a more well-known method used in the treatment of certain cancers (eg, Thomas, Ann NY Acad Sci. 1995 Dec 29; 770: 34-41; Kolb and Holler, Stem Cells. 1997; 15 Suppl 1: 151-8; Thomas, Semin Hematol. 1999 Oct; 36 (4 Suppl 7): 95-103).

          Ln-5 modulators are anti-cancer sonic and shock wave treatments (see, eg, Kambe et al., Hum Cell. 1997 Mar; 10 (1): 87-94); anti-cancer hyperthermia (eg, US Pat. No. 6, , 690, 976) and / or anti-cancer nutritional food therapy (eg, Roudebush et al., Vet Clin North Am Small Anim Pract. 2004 Jan; 34 (1): 249-69, viii and Rafi, Nutrition 2004 Jan; 20 (1): 78-82) and can be delivered in conjunction with the application of other therapeutic methods. Other methods include dietary (eg fasting therapy (may be assisted by anti-obesity agents or appetite suppressants)) or high potassium free of fat or oil and rich in fresh raw fruits and vegetables. , Stealing a low sodium (unsalted) diet (see, for example, A Cancer Therapy: Results of Fifty Cases, Max Gerson, Gerson Inst; 6th edition). Another technique that can be combined with an Ln-5 modulator anti-cancer therapy is an insulin-enhanced therapy where a low dose of insulin is given in combination with a low dose of chemotherapy and an Ln-5 modulator anti-cancer therapy.

          Ln-5 modulator anti-cancer methods are designed to reduce cancer-related and cancer treatment-related symptoms such as depression treatment, pain treatment (eg, by delivery of morphine or morphine derivatives), incontinence treatment, sexual disability treatment, etc. It can also be applied in combination with various additive therapies that have been made.

          In addition or alternatively, the methods of the invention described herein can include one or more agents, related compounds, or combinations thereof that facilitate access of Ln-5 modulators within a tumor. Can be implemented in connection with delivering to Thus, for example, such methods can be performed with the delivery of relaxin, which can relieve the tumor (see, eg, US Pat. No. 6,719,977). As another example of such a technique, an Ln-5 modulator can bind to a cell penetrating peptide (CPP). Cell penetrating peptides and related peptides (such as modified cell penetrating antibodies) are described in, for example, “Zhao et al., J Immunol Methods. 2001 Aug 1; 254 (1-2): 137-45; Hong et al. , Cancer Res. 2000 Dec 1; 60 (23): 6551-6; Lindgren et al., Biochem J. 2004 Jan 1; 377 (Pt 1): 69-76; Buerger et al., J Cancer Res Clin Oncol. 2003 Dec; 129 (12): 669-75; Pooga et al., FASEB J. 1998 Jan; 12 (1): 67-77; and Tseng et al., Mol Pharmacol. 2002 Oct; 62 (4): 864 -72 ". Intracellular administration of a vector comprising a nucleic acid sequence encoding L5G2D3BPn or L5G2D3BP or a related molecule can additionally or alternatively be used to facilitate the therapy of the present invention. .

          Molecules that are involved in aspects of cancer progression that are affected by additional target Ln-5 binding molecules and Ln-5 and thus are a favorable target for a second molecule for combination compositions and / or combination delivery methods include α6β1 Integrin, α3β1 integrin, α2β1 integrin, α6β1 integrin, laminin-6, laminin-7, EGF-R, type VII collagen, fibulin-1, fibulin-2, Rho GTPase, BP180, syndecan-4, nidogen-1, phosphorus Oxidized hsp-27, p300, cytokeratin and other matrix metalloproteinases (eg MMP-1, MMP-2, MMP-9, MMP-10 and MMP-14) (membrane matrix metalloproteinase 1 (MT1) Also known as) Tissue inhibitor -1 of matrix metalloproteinases (TIMP-1) and TIMP-2, E- cadherin, include bone morphogenetic protein -1 (BMP-1) and 67kDa laminin receptor. Thus, in one aspect, the present invention is able to detect Ln-5 modulators and antibodies specific for one or more of these dissimilar molecules in a patient with detectable cancer progression. A method for reducing aspects of cancer progression (eg, cancer cell migration) in a human patient in need of reduction of aspects of cancer progression comprising delivering to a human patient under an amount and under conditions such as provide. Additional types of such molecules are discussed elsewhere herein and / or are known in the art.

          In other aspects, the Ln-5 associated antibody and / or Ln-5 binding protein (or other Ln-5 modulator) is a low molecular weight heparin, standard heparin, pentasaccharide, thrombin inhibitor (melagatran, ximelagatran, etc. And / or thrombosis regulators such as coagulation factors such as Factor VII and Factor VIII. In another aspect, the anticancer agent is an anti-VEGF agent (eg, an anti-VEGF agent such as avastatin), angiostatin, canstatin, combrestatin, endostatin, NM-3, thrombospondin, tumastatin, 2- Antiangiogenic agents such as methoxyestradiol and vitaxin, or interferon α, an inhibitor of hypoxia-inducible factor 1, IL-12, marimastat, neobasstat, angiozyme, thalidomide, squalamine and the like. In yet another aspect, the anticancer agent is an estrogen receptor modulator (SERM) such as tamoxifen, idoxifene, fulvestrant, droloxifene, toremifene, raloxifene, diethylstilbestrol, ethinylestradiol / estinyl. Anticancer agents include: aromatase inhibitors such as anastrazol / arimidex, aminoglutethimide / citradene, exemestane; antiandrogens such as flutamide / eulexin; hydroxyprogesterone caproate, medroxyprogesterone / provera, megestrol acepeto / Progestins such as Megges; corticosteroids such as hydrocortisone and prednisone; luteinizing hormone-releasing hormone (LHRH) analogs such as buserelin and goserelin; hormone inhibitors such as octreotide / sandstatin; cepecitabine / xeloda (Xeloda), cytarabine / Ara- C, cladribine / leustatin, fludaline / fludara, fluorouracil / 5-FU, gemcitabine / gemzar, mercaptopurine / Growth inhibitors such as anti-metabolites such as MP, methotrexate / MTX, thioguanine / 6-TG, allopurinol / zyloprim; busulfan, cyclophosphamide, mechlorethamine, melphalan, thiopeta, semplatin, carboplatin, cisplatin, procarbazine, Acylating agents such as dacarbazine, altretamine, lomustine, carmustine, chlorambucil; topotecan, topoisomerase inhibitors such as camptothecin as irinotecan; podophyllotoxin as etoposide / VP16, teniposide / VP26, etc .; vincristine, vinblastine, vinorelbine Inhibitor of microtubule and / or spindle formation such as taxane, docetaxel, combrestatin, epothilone B as paclitaxel To be possible.

          Anticancer agents include antibiotics such as bleomycin, dactinomycin / actinomycin D, daunorubicin / servidin and anthracycline as doxorubicin, idarubicin, mitomycin, pricamycin; telomerase inhibitors; differentiation inducers; all-trans retinoic acid, 13- Retinoic acid analogs such as cis retinoic acid; Vitamin D analogs such as seocalcitol; Inhibitors of receptor tyrosine kinases; ErbB-1 / EGFR inhibitors such as Iressa and Erbitux; Erb− such as Herceptin 2 / Her2 inhibitors; inhibitors of c-kit such as Gleevec, inhibitors of VEGF receptors such as ZD6474, SU6668; ErbB3, ErbB4, IGF-IR, insulin receptor, PDGFRa, PDGF Inhibitors such as β, Flk2, Flt4, FGFR1, FGFR2, FGFR3, FGFR4, TRKA, TRKC, c-met, Ron, Sea, Tie, Tie2, Eph, Ret, Ros, Alk, LTK, PTK7; Cancer-related enzyme inhibitors such as metalloproteinase inhibitors such as Stat are also included. Other anticancer agents include cathepsin B, a modulator of cathepsin D dehydrogenase activity, glutathione-S-transferases such as glutasylcysteine synthetase and lactate dehydrogenase.

          In other aspects, viral delivery (usually replication deficient) with modified viruses can be used as a delivery vehicle. For example, to deliver a gene encoding a tumor suppressor gene such as human recombinant wild-type p53 / SCH58500, Apc, PTEN, or a gene encoding a death receptor ligand such as TRAIL, TNF, adenovirus, measles Viruses and other viruses can be used. Viral delivery vehicles to deliver oncogenic genes, mutated genes or antisense nucleic acids against deregulated genes, Ras, siRNA against mutated or deregulated genes such as carcinoembryonic antigen (CEA) Can also be used. In addition, delivery of genes encoding interleukins that induce immune responses such as TNF, IL-2, etc. can be performed using such vehicles.

          In a further aspect, the anticancer agent is an HSP90 inhibitor such as 17-allylaminogeldanamycin; an antibody directed against a tumor antigen such as PSA, CA125, KSA; an integrin such as integrin β1; and an inhibitor of VCAM It can be. Other anti-cancer agents include mycophenolate mofetil, mycophenolic acid, asparaginase, calcineurin inhibitor, TOR inhibitor, and the like, and inhibitors of “lymphocyte homing” mechanisms such as FTY720, and adhesion molecule inhibitors (eg, anti-LFA And the like having an effect on cell signaling.

          One or more functions of Ln-5 and / or one or more functions of one or more biomolecules associated with Ln-5 (in an applicable patient or host in a normal and / or associated medical condition) Treatment and / or prevention of these and other diseases and disorders by administering Ln-5 modulators under conditions that are at least partially (eg, at least substantially or substantially completely) disabled. The method of facilitating is another important feature of the present invention.

          The invention further includes distributing information relating to the use of the compound in the prevention or treatment of any of the preceding aspects or any of the symptoms or combinations of symptoms described elsewhere herein. Provided are methods for promoting the sale and / or use of a compound described in any of the preceding aspects or elsewhere herein.

Pharmacological method
Animal and cellular models of IBD or polycystic kidney (PKD) are identified herein as specific Ln-5-associated antibodies, Ln-5 binding proteins or other Ln-5 modulators or various combinations thereof. Can be useful for assessing the effectiveness in promoting, enhancing and / or inducing the desired response discussed in.

For example, colon inflammation can be induced in a mouse, rat, or another mammalian model by known techniques, and macroscopic damage reflects the amount of ulceration and / or inflammation in the mammalian colon. The score can be determined by such techniques in the control and test groups as a means of assessing the ability of the drug to reduce IBD. Other similarly known animal models measure colon strength and / or colon tissue stiffness as a means of evaluating a particular treatment or prevention regime. Specific models include the trinitrobenzene sulfonic acid (“TNBS”) model for IBD. The TNBS model is one of the standard IBD models used in IBD discovery studies and is widely evaluated in rodents. For example, see "CO Elson et al. (1995), Experimental Models of Inflammatory Bowel Disease, Gastroenterology, 109: 1344-1367" and references cited in this document. Another specific model is the dextran sulfate (“DSS”) model. Similar to the TNBS model, colitis induced by DSS is widely used as a screening tool for IBD treatment. Another method for assessing IBD is the use of a chemiluminescent assay for mucosal reactive oxygen metabolites. Eosinophil-associated neurotoxins can also be markers for IBD (see, eg, US Pat. No. 5,928,883). The TNO pharmacological model of inflammatory bowel disease has excellent reproducibility and low mortality compared to the model described. This has been achieved by sensitizing the animals prior to rectal administration of TNBS. The model has been successfully validated using budesonide and sulfasalazine, two of the most commonly used pharmaceutical compounds for the treatment of IBD. Anti-Saccharomyces cerevisiae antibody (ASCA), as well as perinuclear (P-ANCA) or anti-neutrophil cytoplasmic antibody (ANCA) exhibiting an atypical pattern, may be useful in the evaluation of IBD. Commercial systems such as the Prometheus system, IBD FIRST STEP, and IBD DIAGNOSTIC SYSTEM (Promethe Laboratories, Inc.-San Diego, Calif.) Can also be used to evaluate IBD.

          Thus, selection of therapeutic Ln-5 modulators for the treatment of IBD, such as LN-5 binding antibodies useful for IBD treatment, does not use reduced staining of the normal basement membrane of skin or blood vessels. Or can be selected by immunohistochemistry (IHC) detection staining of IBD / disease sites. It is also possible to induce IBD in animals, preferably mice or rats, by adding sodium dextran sulfate (DSS) to drinking water. LN-5-associated antibodies, Ln-5 binding proteins and / or other Ln-5 modulator compositions are preferably administered to animals by injection, but are not limited to injection. Clinical signs of IBD and biochemical markers are recorded to allow discrimination between the composition and the effect of vehicle treatment.

          In another example, IBD is induced in animals, preferably mice or rats, by rectal injection of 2,4,6-trinitrobenzenesulfonic acid (TNBS) solubilized in ethanol. The LN-5 modulator (eg, LN-5-associated antibody, Ln-5 binding protein) is preferably administered to the animal by injection, but is not limited to injection. Clinical signs of IBD and biochemical markers are recorded to allow discrimination between the composition and the effect of vehicle treatment.

In an exemplary aspect, the effects of Ln-5 modulators are evaluated in a model in which IBD is induced by adoptive transfer. Adoptive transfer of CD4 + CD25-T cells to immunodeficient mice (SCID or RAG-KO) induces clinically expressed colitis by two and a half weeks after transfer. Colitis is clinically characterized by weight loss, diarrhea and bloody stool. The macroscopic appearance of the large intestine is conspicuous due to severe enlargement of the intestinal wall that can be observed by histology. Three to five weeks after transmission, inflammation occurs following hypertrophy, but ulcers are only seen in the late stages. Usually, dysplasia or metaplasia is not seen. Lesions are mainly confined to the large intestine (including the cecum) and there are no histological signs of gastritis, pancreatitis or salivary adenitis. In order to examine the potency against IBD of functional Ln-5 modulators with physiological effects, CD4 + CD25-T cells are purified from the spleens of syngeneic donor mice by MACS (Magnetic Activated Cell Sorting). Each mouse is injected intraperitoneally with 3 × 10 ⁵ CD4 + CD25− T cells in a volume of 200 μL RPMI 1640 plus glutamax. Mice are weighed twice a week to check for signs of illness. The Ln-5 modulator is administered via a route that is compatible with optimal bioavailability, such as intravenous, oral, intraperitoneal, subcutaneous, or other suitable route. If necessary, the Ln-5 modulator can be administered repeatedly for an appropriate period of time. For example, the frequency of administration can vary from one or more times daily to every other day, every third day, every week, or every month, and interval treatments can be used as appropriate. In one embodiment, in the case of antibody LN-5 modulators, a dose of 20 mg / kg may be administered twice weekly intraperitoneally in PBS. To assess the prophylactic effect or prevention or delay in a disease or disease symptom, the LN-5 modulator can be administered prior to adoptive transfer. Alternatively, start Ln-5 modulator dosing one to three weeks after adoptive transfer to measure the efficacy of Ln-5 modulator therapy on disease progression, disease development, or disease severity Is possible. The animals are then treated for two and a half to six weeks before sacrifice. Prior to sacrifice, 100 mg / kg BrdU is injected intraperitoneally into 200 μL NaCl physiological solution. At necropsy, the entire gastrointestinal tract is removed for dissection. Before preparing the tissue for immunohistochemistry, the weight, length and width of the gastrointestinal tract are measured. Appropriate physiological effects include significant normalization of the animal's body weight and / or a reduction in the incidence of bloody stool and / or restoration to normal intestinal weight, length and / or thickness and / or significant growth. Reduction (BrdU staining) and / or significant normalization of mucin production (determined by PAS or Alcian blue or Alcian blue HID staining) and / or preservation of goblet cells. Recovery or reduction of inflammation in the IBD state is also a physiological effect that determines effective LN-5 modulators.

          In another exemplary aspect, the effects of Ln-5 modulators are evaluated in a mouse model in which colitis is induced by DSS. Briefly, 8-week old male wild-type C57B1 / 6 mice are treated for 8 days with or without DSS added to their drinking water at a concentration of 1.5%. For example, in the first preliminary experiment, the concentration of DSS was titrated from 1-5%, given to wild type mice, and the dose that caused disease after 8 days was selected for subsequent experiments. Titration of the DSS dose is important. Since the rate of disease induction depends on the type and amount of microorganisms in a particular site, even a change in position can affect the results. After 6-8 days, the mice begin to lose weight. The mice are then weighed twice a week to check for any signs of illness. The potency of a functional Ln-5 modulator having a physiological effect on IBD is compatible with optimal bioavailability and is intravenous, oral, intraperitoneal, subcutaneous or other suitable route. It can be determined by administering the compound by any possible route. If necessary, the Ln-5 modulator can be administered repeatedly for an appropriate period of time. For example, the frequency of administration can vary from one or more times daily to every other day, every third day, every week, or every month, and interval treatments can be used as appropriate. In one embodiment, in the case of antibody LN-5 modulators, a dose of 20 mg / kg may be administered twice weekly intraperitoneally in PBS. The first dose can be administered the day before DSS is added to the drinking water, and administration can continue for 8 to 15 days. Prior to sacrifice, 100 mg / kg BrdU is injected intraperitoneally into 200 μL NaCl physiological solution. At necropsy, the entire gastrointestinal tract is removed for dissection. Before preparing the tissue for immunohistochemistry, the weight, length and width of the gastrointestinal tract are measured. Physiological effects indicative of treatment include significant normalization of the animal's body weight and / or reduction in the incidence of bloody stool and / or restoration and / or proliferation to normal bowel weight, length and / or thickness. Significant reduction (BrdU staining) and / or significant normalization of mucin production (determined by PAS or Alcian blue or Alcian blue HID staining) and / or preservation of goblet cells. Recovery or reduction of inflammation in the IBD state is also a physiological effect that determines effective Ln-5 modulators. Expression of Ln-5 in the DSS model is described by “Pirila et al., Dig Diseases sc. 2003; 48: 93-98”.

The efficacy of a functional Ln-5 modulator with a prophylactic effect that reduces or eliminates increased risk of cancer in IBD patients has been demonstrated in the Apcmin model, a mouse model of multiple intestinal neoplasia (min). It can be detected in mice by assessing the effect on adenomatous polyps. Inbred B6-Apcmin mice can be obtained, for example, from Jackson Laboratory and can be screened for Min / + genotype using a PCR assay. Divide mice into groups that have standardized male and female distribution to avoid clustering of mice from a single litter. Mouse health is monitored every 3 days and weighed weekly. Treatment begins at 28 days of age (at the time of weaning) and continues for several weeks up to 100 days as needed. The compound is compatible for optimal bioavailability and is administered by a route that can be intravenous, oral, intraperitoneal, subcutaneous, or other suitable route. If necessary, the Ln-5 modulator can be administered repeatedly for an appropriate period of time. For example, the frequency of administration can vary from one or more times daily to every other day, every third day, every week, or every month, and interval treatments can be used as appropriate. In an exemplary embodiment, the LN-5 modulator is an antibody and a dose of 20 mg / kg can be administered intraperitoneally in PBS twice a week. Animals are sacrificed when the mice are 90-115 days old at the end of the treatment period. At necropsy, the entire gastrointestinal tract is removed for dissection. The small intestine is divided into three segments of approximately equal length and the large intestine is left untreated. All four intestinal segments are opened longitudinally and washed thoroughly with PBS to remove intestinal contents. Tissues are fixed (60% methanol, 30% chloroform, 10% acetic acid) and rinsed (70% ethanol) prior to tumor counting. Tumor size (area mm ² ) is measured using image analysis of photographed intestinal tissue placed on a calibrated sample stage micrometer. The smallest tumor to be counted has a counting diameter of 0.2 mm. Statistically significant (P <0.05) tumor number and / or tumor size changes are obtained for effective LN-5 modulators.

          As mentioned above, animal models of ADPKD are known and described, for example, by “Thomson et al. (2003 Nov; 285 (5): F870-80)”. Furthermore, the effects of LN-5 modulators on the development and progression of renal cystic disease are described in, for example, “Gattone et al. (2003) Nature Med. 9 (10): 1323-1326” or “Qian et al. (2004). ) J. Biol. Chem (electronic publishing before printing) "can be established in animal models. Establishing the functionality of an Ln-5 modulator based on the administration of an effective amount of an LN-5 modulator to a mouse or rat (depending on the model) and through the development or progression of the disease over 3 to 30 weeks. it can. Appropriate indicators of therapeutic efficacy include renal fibrosis volume and / or plasma BUN (in blood) combined with or reduced by renal cAMP (pmol / mg wet tissue) or decreased renal cyst number or size Urea nitrogen). In one embodiment, two PKD models are examined. One PKD model is the rat, in which splice mutations cause disease by causing a frameshift of the gene Pkhd1, a rat homologous species of PKHD1. Another PKD model is the pcy mouse. Animals obtained from both models have abnormal urinary concentrations and significantly higher kidney cAMP. In a typical assay, animals are monitored for health twice a week and weighed weekly. Urine is collected during the experiment to determine osmolarity. Administration of Ln-5 modulators to PCK rats begins by 3 weeks of age and continues until 10 weeks of age. In the case of pcy mice, treatment begins by 4 weeks of age and continues until 30 weeks of age. The Ln-5 modulator is compatible with optimal bioavailability and is administered by a route that can be intravenous, oral, intraperitoneal, subcutaneous, or other suitable route. If necessary, the Ln-5 modulator can be administered repeatedly for an appropriate period of time. For example, the frequency of administration can vary from one or more times daily to every other day, every third day, every week, or every month, and interval treatments can be used as appropriate. When the Ln-5 modulator is an antibody, a typical treatment regime may be administered twice weekly using a dose of 20 mg / kg administered intraperitoneally in PBS. At the end of the experiment, the animals are sacrificed. Two hours prior to sacrifice, 100 mg / kg BrdU is injected intraperitoneally into 200 μL NaCl physiological solution. At necropsy, blood is collected by cardiac puncture for analysis and the kidney is removed for dissection. Examine kidney weight and visual appearance. The left kidney is fixed in 10% formaldehyde in phosphate buffer (pH 7.4) embedded in paraffin for histological studies. To measure cAMP, the right kidney is immediately frozen in liquid nitrogen. Histological studies use H & E staining to measure cyst volume and Picrosirius red collagen staining to measure fibrosis, both combined with image analysis. Apoptosis index is measured by TUNEL assay and mitotic index is measured by BrdU staining. Blood is analyzed for blood urea nitrogen and serum creatinine. The right kidney is minced in a homogenizer in a 10-fold dose of cold 5% trichloroacetic acid. After centrifugation at 600 × g for 10 minutes, the supernatant is extracted with ether saturated with 3 volumes of water. After drying, the extract is reconstituted in buffer for analysis by enzyme immunoassay using a kit (Sigma-Aldrich). The physiological effect of an effective LN-5 modulator can be seen as a significant decrease in renal accumulation of cAMP measured as pmol / mg wet kidney or pmol / mg protein. A significant (p <0.05) decrease in renal cysts and / or fibrosis and / or kidney weight and / or kidney growth and / or kidney apoptosis index is due to urinary osmolality and / or blood urea nitrogen and / or Or as a normal indicator of useful L-5 modulators, as well as normalization of serum creatinine.

          These methods provide techniques for identifying / screening for Ln-5 modulators with superior anti-IBD and / or anti-ADPKD properties, such as Ln-5-associated antibodies and / or Ln-5 binding proteins.

          Thus, the present invention further provides for a composition comprising an Ln-5-associated antibody, an Ln-5 binding protein, or both to respond in a model of IBD that predicts a correlated therapeutic or prophylactic effect in a mammal. And the therapeutic and / or prophylactic potential of the composition by subjecting the composition to analysis by the model, and assessing the effect of the composition in the model. There is provided a method of screening a candidate composition for biological activity associated with prevention and / or treatment of IBD in a mammal comprising assessing ability.

          In a further aspect, the present invention results in a composition comprising Ln-5-associated antibody, Ln-5 binding protein or both in a model of ADPKD that predicts a correlated therapeutic or prophylactic effect in a mammal. And the therapeutic and / or prophylactic potential of the composition by subjecting the composition to analysis by the model, and assessing the effect of the composition in the model. There is provided a method of screening a candidate composition for biological activity associated with reduced propagation and / or proliferation of renal cysts in a mammal comprising assessing ability.

          In a further aspect, the present invention relates to a composition comprising Ln-5-associated antibody, Ln-5 binding protein or both in a model of irritable bowel syndrome that predicts a correlated therapeutic or prophylactic effect in a mammal. Giving the composition in an amount that is expected to produce a response; subjecting the composition to analysis by the model; and evaluating the effect of the composition in the model by treating the composition and / or A method of screening a candidate composition for biological activity associated with the treatment and / or prevention of irritable bowel syndrome, comprising assessing prophylactic potential.

          The present invention further provides a composition comprising a combination of an Ln-5-associated antibody, Ln-5 binding protein or both and at least one second anticancer agent to correlate therapeutic or prophylactic effects in a mammalian host. In a model that predicts a response in a predicted amount, obtaining a cumulative effect predicted for the combination, subjecting the composition to analysis by the model, and an anti-cancer agent Screening a candidate synergistic combination of anti-cancer agents to identify a synergistic combination comprising assessing whether the combination results in a result that is substantially greater than the cumulative effect expected for the combination Provide a way to do it.

Pharmaceutical composition
In yet another aspect, the invention provides (a) a composition described in any of the preceding aspects or elsewhere herein, and (b) a pharmaceutically acceptable carrier, vehicle, excipient. Shapes, diluents, preservatives, stabilizers, binders, flavoring agents, antioxidants, colorants, adjuvants, disintegrants, solvents, solubilizers, suspending agents, isotonic / isotonic agents , Buffering agents, soothing agents, or any combination thereof (overlapping these, including, for example, two diluents) and (c) directed by a government agency that regulates the manufacture, use, or sale of pharmaceuticals A notice attached to the container in a structured form, to a human or animal to treat at least one symptom described in any of the preceding aspects or a symptom or disease described herein. Notification that is a reflection of the approval of the agency of the pharmaceutical product to be administered. Including in response to provides a pharmaceutical product.

          The compounds of the present invention may be used alone or in combination with other active agents described herein and / or in combination with pharmaceutically acceptable carriers or excipients in single or multiple doses. Can be administered. The pharmaceutical compositions of the present invention are prepared according to conventional techniques such as those disclosed in “Remington: The Science and Practice of Pharmacy, 19th Edition, Gennaro, Ed., Mack Publishing Co., Easton, PA, 1995”. Can be formulated with pharmaceutically acceptable carriers or diluents and any other known adjuvants and excipients.

          The pharmaceutical compositions are oral, rectal, nasal, pulmonary, topical (including buccal and sublingual), transdermal, intracapsular, intraperitoneal, vaginal and parenteral (subcutaneous, intramuscular, intrathecal, intravenous And intradermal) and can be specifically formulated for administration by any suitable route, such as oral, parenteral and intraperitoneal routes being preferred. It will be appreciated that the preferred route will depend on the general condition and the age of the subject to be treated, the nature of the condition to be treated and the active ingredient chosen.

          Pharmaceutical compositions for oral administration include solid dosage forms such as hard or soft capsules, tablets, troches, dragees, pills, troches, powders and granules. Where appropriate, coatings such as enteric coatings can be used to prepare pharmaceutical compositions or, according to methods well known in the art, for active ingredients such as sustained or sustained release. The pharmaceutical composition can be formulated to provide controlled release. Liquid dosage forms for oral administration include solutions, emulsions, aqueous or oily suspensions, syrups and elixirs.

          Pharmaceutical compositions for parenteral administration include sterile aqueous and non-aqueous injectable solutions, dispersions, suspensions or emulsions and sterile powders that should be returned to sterile injectable solutions or dispersions before use. . Injectable depot forms are also contemplated as being within the scope of this invention. Other suitable dosage forms include suppositories, sprays, ointments, creams, gels, inhalants, skin patches, implants and the like.

          A typical oral dosage is about 0.001 to about 100 mg / kg body weight / day, preferably about 0.01 to about 50 mg / day, administered in one or more doses, such as 1 to 3 doses. kg body weight / day, more preferably about 0.05 to about 10 mg / kg body weight / day. The exact dosage will depend on the frequency and mode of administration, sex, age, weight and general symptoms of the subject being treated, the nature and severity of the symptoms being treated, and the complications to be treated, as well as those skilled in the art. Will depend on other factors.

          The formulations can be conveniently given in unit dosage form by methods known to those skilled in the art. Typical unit dosage forms for oral administration one or more times per day, such as 1 to 3 times / day, are 0.05 to about 1000 mg, preferably about 0.1 to about 500 mg, more preferably about 0.5 mg to Can contain about 200 mg.

          For parenteral routes such as intravenous, intrathecal, intramuscular and similar administration, typically the dosage is approximately half of the dose used for oral administration.

          The compounds of the invention are generally used as the free substance or as a pharmaceutically acceptable salt thereof. Examples are acid addition salts of compounds with free base utility and compound base addition salts with free acid utility. The term “pharmaceutically acceptable salt” is generally prepared by reacting the free base with a suitable organic or inorganic acid or by reacting the acid with a suitable organic or inorganic base. Represents non-toxic salts of the compounds of the invention. When a compound of the present invention contains a free base, such salts can be prepared in a conventional manner by treating a solution or suspension of the compound with a chemical equivalent of a pharmaceutically acceptable acid. It is prepared with. Where a compound of the invention contains a free acid, such salts can be prepared in a conventional manner by treating a solution or suspension of the compound with a chemical equivalent of a pharmaceutically acceptable base. It is prepared with. Physiologically acceptable salts of a compound having a hydroxyl group include the anion of the compound in combination with a suitable cation such as sodium or ammonium ion. Other pharmaceutically unacceptable salts may be useful in the preparation of the compounds of the invention, which form a further aspect of the invention.

          For parenteral administration, sterile aqueous solutions, aqueous propylene glycol solutions or solutions of the novel compounds of formula (I) in sesame oil or peanut oil may be used. Such an aqueous solution should be appropriately buffered if necessary. First, the liquid diluent is made isotonic with sufficient physiological saline or glucose. Aqueous solutions are particularly suitable for intravenous, intramuscular, subcutaneous and intraperitoneal administration. All sterile aqueous solvents used are readily available by standard techniques known to those skilled in the art.

          Suitable pharmaceutical carriers include inert solid diluents or fillers, sterile aqueous solution and various organic solvents. Examples of solid carriers are lactose, gypsum, sucrose, cyclodextrin, talc, gelatin, agar, pectin, acacia, magnesium stearate, stearic acid and lower alkyl ethers of cellulose. Examples of liquid carriers are syrup, peanut oil, olive oil, phospholipids, fatty acids, fatty acid amines, polyoxyethylene and water. Similarly, the carrier or diluent may include any sustained release material known in the art, such as glyceryl monostearate or glyceryl distearate, alone or mixed with a wax. The pharmaceutical compositions formed by combining the novel compounds of this invention and pharmaceutically acceptable carriers are then readily administered in a variety of dosage forms suitable for the disclosed route of administration. The formulation can be conveniently given in unit dosage form by methods known in the pharmaceutical arts.

          Formulations of the present invention suitable for oral administration can be presented as discrete units, such as capsules or tablets, each containing a predetermined amount of the active ingredient and which can contain suitable excipients. Furthermore, the orally utilized formulations may be in the form of powders or granules, aqueous or non-aqueous liquids or solutions or suspensions in oil-in-water or water-in-oil liquid emulsions. Compositions intended for oral use can be prepared according to any known method, and such compositions are sweetened to provide a pharmaceutically elegant and palatable preparation. One or a plurality of agents selected from the group consisting of agents, flavoring agents, coloring agents and preservatives can be contained. Tablets contain the active ingredient in admixture with non-toxic pharmaceutically acceptable excipients that are suitable for the manufacture of tablets. These excipients include, for example, inert diluents such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents such as corn starch or alginic acid; binders such as starch, Gelatin or acacia; and lubricants such as magnesium stearate, stearic acid or talc. The tablets may be uncoated or they may be coated by known techniques to delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer period. For example, a time delay material such as glyceryl monostearate or glyceryl distearate may be employed. Tablets are disclosed in US Pat. Nos. 4,356,108, 4,166,452, and 4,265,874 (incorporated herein by reference) to form sustained release osmotic therapeutic tablets. It can also be coated by the technique described in.

          Oral formulations are prepared as hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent such as calcium carbonate, calcium phosphate or kaolin, or the active ingredient is water or an oil medium such as peanut oil, liquid paraffin or It can also be provided as a soft gelatin capsule mixed with olive oil. Aqueous suspensions may contain the active compounds in admixture with excipients suitable for the manufacture of aqueous suspensions. Such excipients are suspending agents, for example sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum arabic; dispersants or wetting agents are naturally occurring such as lecithin Phosphatides or condensation products of alkylene oxides with fatty acids such as polyoxyethylene stearate or condensation products of ethylene oxide with long chain fatty acid alcohols such as heptadecaethyleneoxysetanol, or fatty acid-derived partial esters and monostearic acid Condensation products of ethylene oxide with hexitols such as polyoxyethylene sorbitol, or condensation products of ethylene oxide with fatty acid-derived partial esters and hexitol anhydrides, eg If, polyethylene sorbitan monooleate. Aqueous suspensions may also contain one or more coloring agents, one or more flavoring agents and one or more sweetening agents, such as sucrose or saccharin. Oily suspensions may be formulated by suspending the active ingredient in a vegetable oil, for example arachis oil, olive oil, sesame oil or coconut oil, or in a mineral oil such as liquid paraffin. Oily suspensions may contain a thickening agent, for example beeswax, hard paraffin or cetyl alcohol. Sweetening agents such as those mentioned above, and flavoring agents can be added to provide a palatable oral preparation. These compositions can be preserved by the addition of an anti-oxidant such as ascorbic acid. Dispersible powders and granules suitable for preparation of an aqueous suspension by the addition of water provide the active compound in admixture with a dispersing or wetting agent, suspending agent and one or more preservatives. Suitable dispersing or wetting agents and suspending agents are exemplified by those already mentioned. Additional excipients, for example sweetening, flavoring and coloring agents, can also be present. The pharmaceutical composition of the present invention may be in the form of an oil-in-water emulsion. The oily phase can be a vegetable oil, for example olive oil or arachis oil, or a mineral oil, for example liquid paraffin or mixtures of these. Suitable emulsifiers include naturally occurring gums such as gum arabic or tragacanth, naturally occurring phosphatides such as soy lecithin, and esters or partial esters derived from fatty acids and hexitol anhydrides such as sorbitan monooleate, and The condensation product of the partial ester with ethylene oxide, for example, polyoxyethylene sorbitan monooleate can be used. The emulsion can also contain sweetening and flavoring agents. Syrups and elixirs may be formulated with sweetening agents, for example glycerol, polyethylene glycol, sorbitol or sucrose. Such formulations may also contain a demulcent, a preservative and flavoring and flooring agents. The pharmaceutical compositions may be in the form of a sterile injectable aqueous or oleagenous suspension. This suspension may be prepared according to known methods using those suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that can be employed are water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conveniently used as a solvent or suspending medium. For this purpose any bland fixed oil can be employed using synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid find use in the preparation of injectables.

          The composition can also be in the form of suppositories for rectal administration of the compound of the invention. These compositions are solid at room temperature but liquid at rectal temperature and are therefore prepared by mixing the drug with a suitable nonirritating excipient that can melt in the rectum to release the drug. can do. Such materials include, for example, cocoa butter and polyethylene glycol. For topical use, creams, ointments, jellies, solutions or suspensions containing the compounds of the invention are envisioned. In this application, topical application shall include mouth washes and gargles.

          The compounds of the present invention can also be administered in the form of liposome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles and multilamellar vesicles. Liposomes can be formed from a variety of phospholipids, such as cholesterol, stearylamine or phosphatidylcholines.

          In addition, some of the compounds of the present invention can form solvates with water or common organic solvents. Such solvates are also encompassed within the scope of the present invention.

          Thus, in a further embodiment, a compound of the invention, or a pharmaceutically acceptable salt, solvate or prodrug thereof, and one or more pharmaceutically acceptable carriers, excipients or diluents. A pharmaceutical composition is provided.

          If a solid carrier is used for oral administration, the preparation can be tableted, placed in a hard gelatin capsule in powder or pellet form, or in the form of a troche or medicated candy Is possible. The amount of solid carrier can vary widely but will usually be from about 25 mg to about 1 g. If a liquid carrier is used, the preparation may be in the form of a syrup, emulsion, soft gelatin capsule or sterile injectable liquid such as an aqueous or non-aqueous liquid suspension or solution.

          The Ln-5 modulator composition described herein may be an important feature of the present invention per se (ie, apart from the inventive method described herein).

          All references cited in this specification, including publications, patent applications and patents, are noted to be incorporated individually and specifically by reference, the entirety of which is incorporated herein by reference. To the same extent as described in the text, it is incorporated herein by reference.

          In the context of describing the present invention (especially in the context of the following claims), the use of the terms “a” and “an” and [the] and similar directives, unless stated otherwise, Or should be construed to include both singular and plural unless the context clearly dictates otherwise. The terms “comprising”, “having”, “including” and “including” mean open-ended terms (ie, including but not limited to), unless otherwise specified. Must be interpreted.

          The list of value ranges in this specification, unless otherwise specified in this specification, serves only as an abbreviated way of noting each individual value within the range, Each individual value is incorporated herein as if it were listed separately herein. Unless expressly stated otherwise or clearly contradicted by context, approximate values (eg, “about 10”) described herein can be replaced by the corresponding exact values and vice versa. Is also true.

          All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples or exemplary languages (eg, “etc.”) described herein are for the purpose of clarifying the invention only, and unless otherwise stated. It does not limit the range. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the invention.

          Preferred embodiments of this invention are described herein, including the best mode known to the inventors for carrying out the invention. Variations on those preferred embodiments will be apparent to those of ordinary skill in the art upon reading the foregoing description. The inventor (e.g.) expects those skilled in the art to use such variations as appropriate, and the inventor has other aspects than those specifically described herein. It is intended that the present invention be implemented. Accordingly, this invention includes all modifications and equivalents of the subject matter recited in the claims appended hereto as permitted by applicable law. Moreover, any combination of the above-described elements in all possible variations thereof is encompassed by the invention unless otherwise indicated herein or otherwise clearly contradicted by context.

Example 1: IHC immunohistochemical detection of laminin-5 γ2 expression in IBD
This example describes the detection of Ln-5 γ2 chain expression in an animal model of IBD.

Animal model. CD4 + CD25-T cells were purified from the spleens of syngeneic donor mice by MACS (magnetic activated cell sorting). Each mouse was injected intraperitoneally with 3 × 10 ⁵ CD4 + CD25− T cells in a volume of 200 μL RPMI 1640 plus glutamax. Mice are weighed twice a week for signs of any disease.

          Immunohistochemistry: Mice were injected with bromodeoxyuridine (BrdU, 100 mg / kg) 2 hours before sacrifice. The colon was dissected and fixed in paraformaldehyde and embedded in paraffin. Paraffin from colon sections (4 μm) from control SCID and IBD mice was removed in xylene and rehydrated. Antigen recovery via microwave oven in Tris / EDTA buffer, pH = 9, with 0.5% hydrogen peroxide in Tris buffered saline (TBS) for 20 minutes Endogenous peroxidase activity was blocked at room temperature (RT). Endogenous biotin was blocked using a biotin blocking system from DakoCytomation (Glostrup, Denmark).

          For Ln-5 γ2 immunostaining, to reduce non-specific background, incubate in TBS containing 4% casein (trypton casein peptone, Amersco) for 1 hour at room temperature, then 4% casein / Stained overnight at 4 ° C. with 1 μg / mL rabbit polyclonal anti-human Ln-5 γ2 antibody Pad 28 (obtained from Sirpa Salo, Oulo, Finland) diluted in TBS. A secondary antibody, biotinylated donkey anti-rabbit (1: 3000, Jackson), was applied for 1 hour at room temperature. Signal amplification was performed by incubation with horseradish peroxidase conjugated streptavidin and biotinylated tyramide according to the manufacturer (NEN, Perkin Elmer Life Science, Boston, Mass.). Slides were developed using diaminobenzidine and nuclei were counterstained with hematoxylin.

          In the case of BrdU staining, after incubating the slides for 1 hour at room temperature in TBS containing 10% donkey serum and 0.01% Triton-x-100 to reduce non-specific background, Stained overnight at 4 ° C. with mouse monoclonal anti-BrdU antibody M744 (1:50, DakoCytomation) diluted in TBS with 0.01% Triton-X-100, 7% donkey serum and 3% rat serum. . A secondary antibody, biotinylated donkey anti-mouse (1: 3000, Jackson), was applied for 1 hour at room temperature. Signal amplification was performed by incubation with Vectastein ABComplex according to the manufacturer (Vector). Slides were developed using diaminobenzidine-nickel and the nuclei were counterstained with hematoxylin.

          As shown in FIGS. 1 and 2, Ln-5 γ2 immunopositive cells in tissues given IBD were observed in some of the crypt cells or parts of the crypts. Comparison with growth indices by BrdU staining on adjacent tissue sections (FIG. 2A vs. 2B) revealed that Ln-5 γ2 immunopositive was non-proliferative. Ln-5 γ2-immunopositive cells were found to be predominantly non-mucin producing (no vesicles were found in the cells) (FIG. 1A vs. 1B). Therefore, these cells may be more exposed to chemical and / or pathogen injury.

Example 2: Western blot analysis of siRNA against γ2 chain
This example shows that anti-Ln-5 γ2 siRNA reduces the expression of Ln-5 γ2 chain in vitro.

SW480 cells obtained from ATCC were grown in DME: F12 supplemented with 10% fetal bovine serum and periodically checked for the presence of mycoplasma. Cells were seeded at 3 × 10 ⁵ cells / 2 mL in 6-well plates for transfection. Cells were allowed to attach for at least overnight using Lipofectamine 2000 (Life Technologies) before transfection. Liposomes were formed in solvent without serum by mixing 10 μL Lipofectamine 2000 and 3 μg plasmid DNA or 15 μL 20 pmol / μL oligonucleotide in a total volume of 250 μL. The following oligonucleotides were used. # 7 (SEQ ID NO: 7; control) and # 6 (SEQ ID NO: 6; control), and siRNA sequences # 2 (SEQ ID NO: 2), and # 5 (SEQ ID NO: 5) and the entire Ln-5 γ2 chain; Corresponding antisense sequence. In order to prepare the sequence encoding the antisense construct for the Ln-5 γ2 chain, the following sequence was placed in the expression vector pCDNA3.1 + (Invitrogen).

          ccg atc aa gat aca ctg cct gga ctt ccc att gca atc aca gac tt ccc cct gga ggt gg c c c g gc tgc gg g ag g ag

          After liposome formation, this mixture was added to the cells and incubated for 5 hours in a cell incubator before changing the solvent to complete solvent with serum. After 24 hours, cells were analyzed for effects on laminin-5 γ2 expression.

          The solvent was collected and centrifuged. The attached cells were scraped with a rubber policeman into ice-cold PBS. Cell pellets obtained from the solvent were pooled with the scraped cells and centrifuged again (1200 rpm, 3 minutes). The supernatant is drained and the cells are lysed on ice with 20 μL lysis buffer (50 mM Hepes, pH 8.0, 150 mM NaCl, 1% v / v Triton X100, 2 mM EDTA, 10% v / v glycerol). did. Cells were centrifuged at 13,000 × g for 5 minutes and loading buffer was added to a final concentration of 1 × (6.57 μL of 4 × loading buffer). As before, samples were sonicated on ice prior to centrifugation. Prior to loading each lane on a pre-formed gel (NuPAGE 4-12% BisTris), the sample was heated to 90 ° C. for 10 minutes and electrophoresed at 180 V for 1 hour. For sizing and blotting efficiency, 5 μL of rainbow marker and 2 μL of biotin labeled marker were loaded into each lane. During wet blotting, the protein was electroblotted onto a nitrocellulose membrane for 3 hours at 90 V and detected by Western blot.

          To detect an internal standard for protein loading, the membrane was cut into two parts using a rainbow marker and was found to be approximately 50-60 kDa in size. Two immunodetections were performed in parallel using a membrane with a high molecular weight protein for laminin-5 γ2 detection and a membrane with a low molecular weight for actin detection (internal protein control for loading). The membrane was blocked for 1 hour at 25 ° C. (Blocking buffer: 5% non-fat dry milk in PBS, 0.1% tween-20). After blocking, primary antibodies (polyclonal rabbit anti-laminin-5 γ2 1: 1000 or peroxidase-conjugated anti-actin 1: 1000 in PBS supplemented with 0.1% tween-20 and 0.5% non-fat dry milk) Prior to addition, membranes were rinsed once in PBS and incubated overnight (about 16 hours) at 4 ° C. Rinse the membrane 3x5 min with PBS supplemented with 0.1% tween-20, horseradish peroxidase conjugated secondary antibody (for detection of laminin-5 γ2) 0.1% tween-20 and 0.5% With goat anti-rabbit 1: 15,000 in PBS supplemented with non-fat dry milk) and incubated at 25 ° C. for 1 hour. Membranes were rinsed 3 × 5 minutes in PBS supplemented with 0.1% tween-20 prior to visualization with ECL (Amersham) according to the supplier's instructions.

          In FIG. 3, immunoblot detection of laminin-5γ2 is shown at the top from the Western blot of SW480 cell lysate, whereas immunoblot detection of actin is shown below. The immunoblot of laminin-5 γ2 shows two distinct bands corresponding to a long 155 kDa chain and a shorter 105 kDa chain. In all siRNA against laminin-5 γ2, reduced expression is seen mainly on the large form of strands. This long form of laminin-5γ2 is also secreted from the cell and is therefore expected to be the first form affected by siRNA.

Example 3: siRNA cytotoxicity in combination with other anticancer drugs
This example shows that susceptibility to cell death induction is increased by decreasing laminin-5 γ2 expression.

SW480 cells obtained from ATCC were grown in DME: F12 supplemented with 10% fetal bovine serum and periodically checked for the presence of mycoplasma. For transfection, cells were seeded in 6 well plates as 3 × 10 ⁵ cells / 2 mL. Cells were allowed to attach for at least overnight using Lipofectamine 2000 (Life Technologies) before transfection. Liposomes were formed in solvent without serum by mixing 10 μL Lipofectamine 2000 and 3 μg plasmid DNA or 15 μL 20 pmol / μL oligonucleotide in a total volume of 250 μL. After liposome formation, this mixture was added to the cells and incubated for 5 hours in a cell incubator before changing the solvent to complete solvent with serum. After 24 hours, cells were trypsinized and transferred to 96-well plates for analysis for induction of death.

          Cells were seeded for 24 hours (all in triplicate) before dosing the title compound. This compound was solubilized in DMSO and added to cells in medium supplemented with 0.1% v / v or less DMSO. Triplicate cell cultures for basal cell death (untreated) and maximal response (complete lysis) and triplicate media samples for background measurements were included. Cells were incubated for an additional 24 hours. Maximal response (100% cell death) is induced by adding 1% v / v Triton X100 to triplicate cell culture medium 5 minutes before collecting medium for detection of lactate dehydrogenase (LDH) activity did. Media from all samples was used in the detection of LDH using Takara Bio Inc's kit.

          Transfection of SW480 cells with siRNA against γ2 increased spontaneous cell death when compared to SW480 cells transfected with the same length of scrambled RNA. The addition of low doses of thapsigargin, staurosporine, TRAIL or tunicamycin did not alleviate cell death increased by siRNA against γ2. Any combination of anti-cancer compounds was still effective against cell death in SW480 cells.

Exemplary aspects and features of the invention
1. A patient having a significant risk of developing inflammatory bowel disease or identified as at risk is given a composition comprising laminin-5-associated antibody, laminin-5 binding protein, or both, in one or more physiology. Induce one or more physiological responses associated with the treatment or prevention of inflammatory bowel disease, including administering in an amount sufficient to detectably induce, promote and / or enhance an inflammatory response , Promote and / or strengthen.

          2. A patient population identified as having or identified as having a significant risk of developing inflammatory bowel disease is administered a composition comprising laminin-5-associated antibody, laminin-5 binding protein, or both, to a similar population of patients Administration in an amount sufficient to detectably reduce the severity, transmission, onset or risk of developing inflammatory bowel disease in and / or facilitate treatment of inflammatory bowel disease in a similar patient population A method of reducing the severity, transmission, onset or risk of developing inflammatory bowel disease and / or promoting its treatment.

          3. 3. The method of aspect 1 or aspect 2, wherein the method comprises administering to the patient a therapeutically effective amount of laminin-5-binding integrin.

          4). 4. The method of aspect 3, wherein the method comprises administering to the patient a therapeutically effective amount of one or more laminin-5-binding integrins selected from the group consisting of α3β1, α6β1 and α6β4.

          5. 5. The method according to any one of aspects 1 to 4, wherein the method comprises administering to the patient a therapeutically effective amount of an antibody that binds laminin-5.

          6). 6. The method of aspect 5, wherein the laminin-5 antibody detectably binds the γ2 chain of laminin-5.

          7. 6. The method of aspect 5, wherein the laminin-5 antibody detectably binds the α3 chain of laminin-5.

          8). 6. The method of aspect 5, wherein the laminin-5 antibody detectably binds the β3 chain of laminin-5.

          9. 9. The method according to any of aspects 1 to 8, wherein said method comprises administering to said patient a therapeutically effective amount of an antibody that binds laminin-5 binding integrin.

          10. 10. The method of aspect 9, wherein the antibody binds an integrin selected from the group consisting of α3β1, α6β1 and α6β4.

          11. The method according to any of the preceding aspects, wherein said patient suffers from ulcerative colitis.

          12 Administering a composition comprising a laminin-5-associated antibody, laminin-5 binding protein or both to the patient in an amount sufficient to detectably reduce the risk of developing cancer in a population of similar patients. A method for reducing the risk of developing cancer in a patient suffering from inflammatory bowel disease.

          13. In patients suffering from inflammatory bowel disease and patients identified as having a cancer associated with inflammatory bowel disease or at risk of developing cancer associated with inflammatory bowel disease, laminin-5 Relevant to the prevention and / or treatment of cancer in said patient comprising administering a composition comprising an associated antibody, laminin-5 binding protein, or both in an amount sufficient to detectably induce a physiological response. A method for inducing, promoting and / or enhancing a physiological response.

          14 14. The method of aspect 12 or aspect 13, wherein the method comprises administering to the patient a therapeutically effective amount of laminin-5-binding integrin.

          15. 15. The method of aspect 14, wherein the method comprises administering to the patient a therapeutically effective amount of one or more laminin-5-binding integrins selected from the group consisting of α3β1, α6β1, and α6β4.

          15. 15. The method of any one of aspects 12 to 14, wherein the method comprises administering to the patient a therapeutically effective amount of an antibody that binds laminin-5.

          16. 16. The method of aspect 15, wherein the laminin-5 antibody detectably binds the γ2 chain of laminin-5.

          17. 16. The method of aspect 15, wherein the laminin-5 antibody detectably binds the α3 chain of laminin-5.

          18. 16. The method of aspect 15, wherein the laminin-5 antibody detectably binds the β3 chain of laminin-5.

          19. 19. The method according to any of aspects 12-18, wherein said method comprises administering to said patient a therapeutically effective amount of an antibody that binds laminin-5 binding integrin.

          20. 20. The method of aspect 19, wherein the antibody binds an integrin selected from the group consisting of α3β1, α6β1 and α6β4.

          21. 21. A method according to any of aspects 12 to 20, wherein the patient suffers from ulcerative colitis.

          22. The patient is identified as suffering from cancer, and the method comprises administering an anticancer agent, or the patient is identified as being at substantial risk of developing cancer associated with inflammatory bowel disease; The method according to any one of aspects 12 to 21, wherein the method comprises administering a cancer preventive agent to the patient.

          23. To a patient identified as having a renal cyst or a patient identified as having a substantial risk of developing such a cyst, a composition comprising laminin-5-associated antibody, laminin-5 binding protein, or both Said patient in an amount sufficient to detectably reduce the risk of developing a renal cyst in a population of similar patients, detectably prevent the onset, or detectably reduce transmission, proliferation or propagation and proliferation A method of reducing the risk of developing a renal cyst, preventing the onset, or reducing transmission, proliferation or propagation and proliferation in the patient, comprising administering to the patient.

          24. A patient identified as having a renal cyst or having a substantial risk of developing a renal cyst may receive a physiological response with a composition comprising laminin-5-associated antibody, laminin-5 binding protein, or both. A method of inducing, promoting and / or enhancing a physiological response in said patient comprising administering in an amount sufficient to detectably induce, promote and / or enhance.

          25. Renal in said patient comprising administering to a patient having a renal cyst a composition comprising laminin-5-associated antibody, laminin-5 binding protein or both in an amount sufficient to detectably reduce the condition A method of reducing the pathology associated with cyst growth and / or propagation.

          26. 26. The method of any one of aspects 23-25, wherein the method comprises administering to the patient a therapeutically effective amount of laminin-5-binding integrin.

          27. 27. The method of aspect 26, wherein the method comprises administering to the patient a therapeutically effective amount of one or more laminin-5-binding integrins selected from the group consisting of α3β1, α6β1, and α6β4.

          28. 28. A method according to any one of aspects 23 to 27, wherein the method comprises administering to the patient a therapeutically effective amount of an antibody that binds laminin-5.

          29. 29. The method of aspect 28, wherein the laminin-5 antibody detectably binds the γ2 chain of laminin-5.

          30. 29. The method of aspect 28, wherein the laminin-5 antibody detectably binds the α3 chain of laminin-5.

          31. 29. The method of aspect 28, wherein the laminin-5 antibody detectably binds the β3 chain of laminin-5.

          32. 32. The method according to any of aspects 23 to 31, wherein the method comprises administering to the patient a therapeutically effective amount of an antibody that binds laminin-5 binding integrin.

          33. 33. The method of aspect 32, wherein the antibody binds an integrin selected from the group consisting of α3β1, α6β1, and α6β4.

          34. 34. A method according to any of aspects 23 to 33, wherein the method comprises administering at least one second anti-renal cyst agent to the patient.

          35. In order to achieve the desired results, a host suffering from irritable bowel syndrome or identified as being at a substantial risk of developing irritable bowel syndrome has received laminin-5-associated antibody, laminin-5 Reducing the risk of developing irritable bowel disease in a patient, including administering a therapeutically or prophylactically effective amount of a binding protein or both, delaying the onset of irritable bowel disease in a patient, reducing severity; A method for preventing and / or treating inflammatory bowel disease.

          36. A composition comprising a certain amount of laminin-5-associated antibody, laminin-5 binding protein, or both, in an amount sufficient to detectably promote, enhance and / or induce a physiological response. A method of promoting, enhancing and / or inducing one or more physiological effects associated with the prevention and / or treatment of irritable bowel syndrome in said patient.

          37. 37. The method of any one of aspects 35 to 36, wherein the method comprises administering to the patient a therapeutically effective amount of laminin-5-binding integrin.

          38. 38. The method of aspect 37, wherein the method comprises administering to the patient a therapeutically effective amount of one or more laminin-5-binding integrins selected from the group consisting of α3β1, α6β1 and α6β4.

          39. 39. The method of any one of aspects 35 to 38, wherein the method comprises administering to the patient a therapeutically effective amount of an antibody that binds laminin-5.

          40. 40. The method of aspect 39, wherein the laminin-5 antibody detectably binds the γ2 chain of laminin-5.

          41. 40. The method of aspect 39, wherein the laminin-5 antibody detectably binds the α3 chain of laminin-5.

          42. 40. The method of aspect 39, wherein the laminin-5 antibody detectably binds the β3 chain of laminin-5.

          43. 43. The method according to any of aspects 35 to 42, wherein the method comprises administering to the patient a therapeutically effective amount of an antibody that binds laminin-5 binding integrin.

          44. 44. The method of aspect 43, wherein the antibody binds an integrin selected from the group consisting of α3β1, α6β1, and α6β4.

          45. 45. The method of any one of aspects 35 to 44, wherein the method comprises administering at least one second anti-irritable bowel syndrome agent to the patient.

          46. A composition comprising a synergistically effective combination of a laminin-5-associated antibody, laminin-5 binding protein or both and one or more anticancer agents in a mammalian host afflicted with cancer can detect the progression of said cancer A method of reducing cancer progression in a mammalian host afflicted with cancer, comprising administering in an amount sufficient to reduce it.

          47. A composition comprising a synergistically effective combination of laminin-5-associated antibody, laminin-5 binding protein or both and one or more anticancer agents to increase the ratio of quiescent cells to invasive cells in a mammalian host A method of increasing the ratio of quiescent cells to invasive neoplastic cells in a mammalian host comprising administering to the mammalian host a therapeutically effective amount of the product.

          48. Expected to be associated with cancer, detectably reduce the risk of developing a tubular network associated with cancer, detectably prolong the occurrence of tubular network associated with cancer, and / or formed in a mammalian host Therapeutic efficacy of a composition comprising a synergistically effective combination of laminin-5-associated antibody, laminin-5 binding protein or both and one or more anticancer agents to detectably reduce the number of tubular networks A method of inhibiting the formation of a tubular network associated with cancer in a mammalian host comprising administering an amount to said mammalian host.

          49. Therapeutic of a composition comprising a synergistically effective combination of laminin-5-associated antibody, laminin-5 binding protein or both and one or more anticancer agents to detectably reduce the invasive potential of cancer cells A method of reducing the invasive ability of cancer cells in a mammalian host, comprising administering an effective amount to the mammalian host.

          50. In order to achieve the desired result, a therapeutically effective amount of a composition comprising a synergistically effective combination of laminin-5-associated antibody, laminin-5 binding protein or both and one or more anticancer agents is administered to a mammalian host. A method of reducing cell migration, reducing tumor growth, reducing neoplastic or pre-neoplastic cell division, or reducing any combination thereof in a mammalian host, comprising administering to a mammalian host.

          51. Synergistically laminin-5-associated antibody, laminin-5 binding protein or both and one or more anticancer agents to detectably modulate MAP kinase activity in a neoplastic or pre-neoplastic cell of a mammalian host A method of modulating MAP kinase activity in a neoplastic or pre-neoplastic cell of a mammalian host comprising contacting said cell with a physiologically effective amount of a composition comprising an effective combination.

          52. Prophylactic of a composition comprising a synergistically effective combination of laminin-5-associated antibody, laminin-5 binding protein or both and one or more anticancer prophylactic agents to achieve a desired physiological effect Reducing the risk of developing cancer in a mammalian host, including administering an effective amount to the mammalian host, reducing the time to onset of cancerous symptoms, reducing the severity of cancer diagnosed at an early stage, And / or a method of reducing the affected area of cancer at the time of onset of cancer.

          53. 53. The method of any one of aspects 46-52, wherein the method comprises administering to the patient a therapeutically effective amount of laminin-5-binding integrin.

          54. 55. The method of aspect 54, wherein the method comprises administering to the patient a therapeutically effective amount of one or more laminin-5-binding integrins selected from the group consisting of α3β1, α6β1 and α6β4.

          55. 55. The method of any one of aspects 46 to 54, wherein the method comprises administering to the patient a therapeutically effective amount of an antibody that binds laminin-5.

          55. 56. The method of aspect 55, wherein the laminin-55 antibody detectably binds the γ2 chain of laminin-5.

          56. 56. The method of aspect 55, wherein the laminin-5 antibody detectably binds the α3 chain of laminin-5.

          57. 56. The method of aspect 55, wherein the laminin-5 antibody detectably binds the β3 chain of laminin-5.

          58. 58. The method of any of aspects 46-57, wherein the method comprises administering to the patient a therapeutically effective amount of an antibody that binds laminin-5 binding integrin.

          59. 59. The method of aspect 58, wherein the antibody binds an integrin selected from the group consisting of α3β1, α6β1 and α6β4.

          60. Any of the preceding aspects, including disseminating information relating to the use of said compound in the prevention or treatment of any symptom or combination of conditions described elsewhere in this specification Or a method of promoting the sale and / or use of a compound described elsewhere herein.

61. (A) a composition as described in any of the preceding aspects or elsewhere herein; and (b) a pharmaceutically acceptable carrier, vehicle, excipient, diluent, preservative, stable. Agent, binder, flavoring agent, antioxidant, colorant, adjuvant, disintegrant, solvent, solubilizer, suspending agent, isotonic / isotonic agent, buffer, soothing agent, or these Any combination (including any overlap of these, eg, including two diluents);
Including
(C) at least one symptom described in any of the preceding aspects, or a description herein, attached to said container in a form directed by a government agency that regulates the manufacture, use or sale of a medicament; A notification that is a reflection of the approval by the institution of a pharmaceutical product for administration to a human or animal to treat other symptoms or illnesses described in
A pharmaceutical product containing as needed.

62. (A) an amount of a composition comprising a laminin-5 associated antibody, a laminin-5 binding protein, or both, that is expected to produce a response in a model of inflammatory bowel disease that predicts a correlated therapeutic or prophylactic effect in a mammal. And giving in
(B) subjecting the composition to analysis by the model; and
(C) evaluating the therapeutic and / or prophylactic potential of the composition by evaluating the effect of the composition in the model;
A method of screening a candidate composition for biological activity associated with the prevention and / or treatment of inflammatory bowel disease in a mammal.

63. (A) Predicting that a composition comprising a laminin-5 associated antibody, laminin-5 binding protein or both will produce a response in a model of autosomal dominant polycystic kidney disease that predicts a correlated therapeutic or prophylactic effect in mammals Giving in the amount to be
(B) subjecting the composition to analysis by the model; and
(C) evaluating the therapeutic and / or prophylactic potential of the composition by evaluating the effect of the composition in the model;
A method of screening a candidate composition for biological activity associated with reduced propagation and / or proliferation of renal cysts in a mammal.

64. (A) an amount of a composition comprising laminin-5 associated antibody, laminin-5 binding protein, or both, that is expected to produce a response in a model of irritable bowel syndrome that predicts a correlated therapeutic or prophylactic effect in mammals And giving in
(B) subjecting the composition to analysis by the model; and
(C) evaluating the therapeutic and / or prophylactic potential of the composition by evaluating the effect of the composition in the model;
A method of screening a candidate composition for biological activity associated with the treatment or prevention of irritable bowel disease.

65. (A) A composition comprising a combination of a laminin-5 associated antibody, a laminin-5 binding protein or both and at least one second anticancer agent in a model that predicts a correlated therapeutic or prophylactic effect in a mammalian host Giving in the amount expected to bring
(B) obtaining the cumulative effect expected for the combination;
(B) subjecting the composition to analysis by the model; and
(C) evaluating whether the combination yields a result that is substantially greater than the cumulative effect expected for the combination to identify a synergistic combination of anti-cancer agents;
Screening for candidate synergistic combinations of anticancer agents.

          66. 68. The method of aspect 65, wherein the second anticancer agent is selected from the agents described herein.

          67. The method according to any of the preceding aspects, wherein the method is performed using one or more non-Ln-5 associated antibodies, non-Ln-5 binding proteins, Ln-5 modulators.

          68. 68. The method of aspect 67, wherein the Ln-5 modulator is siRNA, an antisense molecule or a small molecule Ln-5 modulator.

          69. A method of treating inflammatory bowel disease, comprising administering a composition comprising an effective amount of a laminin-5 modulator to a subject suffering from inflammatory bowel disease.

          70. A method of reducing the risk of inflammatory bowel disease, comprising administering a composition comprising an effective amount of a laminin-5 modulator to a subject at risk of inflammatory bowel disease.

          71. A method of reducing the risk of inflammatory bowel disease-related colorectal cancer comprising administering a composition comprising an effective amount of a laminin-5 modulator to a subject suffering from inflammatory bowel disease.

          72. The method according to any of aspects 69 to 71, wherein the inflammatory bowel disease is ulcerative colitis.

          73. The method according to any of aspects 69 to 71, wherein the inflammatory bowel disease is Crohn's disease.

          74. 74. A method according to any of aspects 69 to 73, comprising administration of at least one additional anti-inflammatory bowel disease agent.

          75. A method of treating irritable bowel syndrome, comprising administering a composition comprising an effective amount of a laminin-5 modulator to a subject suffering from irritable bowel syndrome.

          76. A method of reducing the risk of irritable bowel syndrome, comprising administering a composition comprising an effective amount of a laminin-5 modulator to a subject at risk of irritable bowel syndrome.

          77. 77. The method of any of aspects 75 and 76, comprising administration of at least one additional anti-irritable bowel syndrome agent.

          78. A method of treating polycystic kidney, comprising administering a composition comprising an effective amount of a laminin-5 modulator to a subject identified as suffering from polycystic kidney disease.

          79. A method of reducing the progression of polycystic kidney, comprising administering a composition comprising an effective amount of a laminin-5 modulator to a subject identified as suffering from autosomal dominant polycystic kidney disease.

          80. 80. The method of any of aspects 78 and 79, comprising administration of at least one additional anti-cystic kidney agent.

          81. A method of treating cancer comprising administering to a subject suffering from cancer a composition comprising a synergistically effective combination of a laminin-5 modulator and at least one second anticancer agent.

          82. A method of reducing tumor growth comprising administering to a subject suffering from a tumor a composition comprising a synergistically effective combination of a laminin-5 modulator and at least one second anticancer agent.

          83. Tumor invasive or metastatic growth comprising administering to a subject suffering from a primary tumor a composition comprising a synergistically effective combination of a laminin-5 modulator and at least one second anticancer agent How to reduce.

          84. Said second anti-cancer agent is melphalan, busulfan, cis-platin, carboplatin, cyclophosphamide, ifosfamide, dacarblamin, dacarbamin Chlorambucil, thiotepa, lomustine, temozolamide, cytarabine, azathioprine, fludarabine phosphate, fludarabine phosphate 5-fluorouracil, gemcitabine, hydroxyurea, cladribine, thioguanine, methotrebin, capecitabine, capecitabine, capecitabine, capecitabine, capecitabine , Docetaxel (Docetaxel), paclitaxel (Paclitaxel), doxorubicin (Axacrubine), amsacrine, irinotecan, daunorubicin (Daunorubicin), epirubicin E icin), mitomycin (Mitomycin), mitoxantrone, idarubicin, teniposide, etoposide, topotecan, thiolamin e ), Polyestradiol phosphate, estradiol, anastrozole, exemestane, letrozole, tamoxifen, megestrol acetic acid (Megerol acetic acid) trol acetate, medroxyprogesterone acetic acid, octreotide, cyproterone acetic acid, bicartimide, flutriline, flutrimine ,), Goserelin, and any other cancer agent described herein, and any combination thereof, The method of any of aspects 81-83.

          85. Any of aspects 69 to 84, wherein said laminin-5 modulator is selected from the group consisting of an antibody that binds laminin-5, a laminin-5 binding protein, siRNA, an antisense molecule or a small molecule Ln-5 modulator. The method described in 1.

          86. 86. The method of aspect 85, wherein the laminin-5 modulator is an antibody that binds laminin-5 γ2 chain.

          87. 86. The method of aspect 85, wherein the laminin-5-modulator is an siRNA derived against a nucleic acid encoding a laminin-5 γ2 chain.

          88. Use of a laminin-5-modulator in the preparation of a medicament for treating inflammatory bowel disease.

          89. Use of laminin-5-modulator in the preparation of a medicament for the treatment of inflammatory bowel disease-related colorectal cancer.

          90. Use of laminin-5-modulator in the preparation of a medicament for treating irritable bowel syndrome.

          91. Also provided is the use of an effective amount of laminin-5-modulator and at least one second anticancer agent in the preparation of a medicament for treating cancer.

          92. Use of an effective amount of laminin-5-modulator and at least one second anticancer agent in the preparation of a medicament for inhibiting tumor growth.

          93. Use of an effective amount of laminin-5-modulator and at least one second anticancer agent in the preparation of a medicament for reducing the risk for metastatic growth of a primary tumor.

          94. Aspects 88-93, wherein the laminin-5 modulator is selected from the group consisting of antibodies directed against laminin-5, laminin-5 binding protein, siRNA, antisense molecule or small molecule Ln-5 modulator. Use as described in any of the above.

          95. 95. Use according to aspect 94, wherein said laminin-5 modulator is an antibody directed against laminin-5 γ2 chain.

          96. 95. Use according to aspect 94, wherein the laminin-5-modulator is a siRNA derived against a nucleic acid encoding a laminin-5 γ2 chain.

          97. Treatment for inflammatory bowel disease, multiple cystic kidney disease or irritable bowel syndrome (wherein the inflammatory bowel disease includes ulcerative colitis and Crohn's disease), including means for modulating Ln-5.

          98. Information regarding the use of Ln-5 in the treatment of inflammatory bowel disease, polycystic kidney disease or irritable bowel syndrome (wherein inflammatory bowel disease includes ulcerative colitis and Crohn's disease) A method of promoting the sale of Ln-5 modulators, including providing to medical professionals and / or organizations including, but not limited to, insurance professionals, research scientists and / or patients.

Paraffin sections of colon obtained from (A) IBD and (B) normal control mice were immunostained for Ln-5 γ2 chain expression. Arrows indicate Ln-5 γ2 immunopositive epithelial cells. Paraffin sections of IBD colon were immunostained for (A) Ln-5 γ2 chain and (B) BrdU. Arrows indicate Ln-5 γ2 immunopositive, BrdU negative epithelial cells. Western blot analysis of the effect of siRNA on Ln-5 γ2 expression in an in vitro model. Cell death of SW480 cells transfected with anti-LN-5 γ2 siRNA and treated with chemotherapeutic compounds.

Claims (19)

          A method of treating inflammatory bowel disease, comprising administering a composition comprising an effective amount of a laminin-5 modulator to a subject suffering from inflammatory bowel disease.           A method of reducing the risk of inflammatory bowel disease-related colorectal cancer comprising administering a composition comprising an effective amount of a laminin-5 modulator to a subject suffering from inflammatory bowel disease.           The method according to claim 1, wherein the inflammatory bowel disease is ulcerative colitis.           The method according to claim 1, wherein the inflammatory bowel disease is Crohn's disease.           A method of treating polycystic kidney, comprising administering a composition comprising an effective amount of a laminin-5 modulator to a subject identified as suffering from polycystic kidney disease.           A method of inhibiting the progression of polycystic kidney, comprising administering a composition comprising an effective amount of a laminin-5 modulator to a subject identified as suffering from autosomal dominant polycystic kidney disease.           A method of treating cancer comprising administering to a subject suffering from cancer a composition comprising an effective amount of a laminin-5 modulator and at least one second anticancer agent.           The laminin-5 modulator is selected from the group consisting of antibodies directed against laminin-5, laminin-5 binding protein, siRNA, antisense molecule or small molecule Ln-5 modulator. 9. The method according to any one of 8.           9. The method of claim 8, wherein the laminin-5 modulator is an antibody induced against laminin-5 γ2 chain.           9. The method of claim 8, wherein the laminin-5-modulator is an siRNA derived against a nucleic acid encoding a laminin-5 γ2 chain.           Use of a laminin-5-modulator in the preparation of a medicament for treating inflammatory bowel disease.           12. Use according to claim 11, wherein the inflammatory bowel disease is ulcerative colitis.           12. Use according to claim 11, wherein the inflammatory bowel disease is Crohn's disease.           Use of laminin-5-modulator in the preparation of a medicament for the treatment of inflammatory bowel disease-related colorectal cancer.           Use of an effective amount of laminin-5-modulator and at least one second anticancer agent in the preparation of a medicament for treating cancer.           Use of an effective amount of laminin-5-modulator and at least one second anticancer agent in the preparation of a medicament for inhibiting tumor growth.           12. The laminin-5 modulator is selected from the group consisting of antibodies directed against laminin-5, laminin-5 binding protein, siRNA, antisense molecule or small molecule Ln-5 modulator. 16. Use according to any of 16.           18. Use according to claim 17, wherein the laminin-5 modulator is an antibody directed against laminin-5 γ2 chain.           18. Use according to claim 17, wherein the laminin-5-modulator is a siRNA derived against a nucleic acid encoding a laminin-5 γ2 chain.

Images