CN1942199A - Methods and compositions for the treatment of cancer and infectious disease using alpha (2) macroglobulin-antigenic molecule complexes - Google Patents

Abstract

The present invention relates to the use of alpha (2) macroglobulin complexes isolated from the serum of a mammal. The invention also relates to methods for making such complexes and compositions comprising alpha (2) macroglobulin complexes, isolated from the serum of a mammal, wherein such compositions are used in methods for the treatment and prevention of cancer and infectious disease. The invention also relates to methods for treating and preventing cancer and infectious disease using such complexes comprising, isolated from the serum of a mammal. The invention also encompasses methods for production of alpha (2) macroglobulin complexes.

Description

Use the method and composition of α (2) macroglobulin-antigenic molecule complexes treatment cancer and infectious disease

The application requires the U.S. Provisional Application No.60/449 of submission on February 20th, 2003, the U.S. Provisional Application No.60/450 that on February 27th, 022 and 2003 submitted to, and 751 priority, described application is all incorporated into this paper as a reference in full.

The present invention finishes under the support of the government-funded CA A184479 that NIH (the National Institutes ofHealth) authorizes.Government enjoys certain right in the present invention.

1. brief introduction

The present invention relates to the application of isolating α (2) macroglobulin complex in the method for treatment and prophylaxis of cancer and infectious disease from mammalian blood serum.The present invention also comprises the method for preparing α (2) macroglobulin complex.The present invention also comprises the compositions and the method for α (2) macroglobulin-antigenic molecule complexes that is used to prepare derived from serum.

2. background of invention

2.1. α (2) macroglobulin

α-macroglobulin is the proteic protein superfamily of structurally associated, and described structurally associated albumen also comprises complement component C3, C4 and C5.Human plasma protein fraction matter α (2) macroglobulin (α 2M) is the equal four polyprotein matter of 720kDa, mainly be called as protease inhibitor and blood plasma and inflammatory fluid protease remove molecule (summary is referring to Chu and Pizzo, 1994, Lab.Invest., 71:792).α 2M synthesizes the precursor with 1474 amino acid residues.Preceding 23 aminoacid play the effect of signal sequence, its fracture obtain having 1451 amino acid residues maturation protein (people such as Kan, 1985, Proc.Natl.Acad.Sci., U.S.A., 82:2282-2286).

α (2) macroglobulin with the mode of covalency mussily (promiscuously) be incorporated into protein and peptide (people such as Chu with nucleophilicity amino acid side chain, 1994, Ann.N.Y.Acad.Sci., 737:291-307) and make their targeting (be also referred to as α 2M receptor or α 2MR in the cell of expressing CD91; Chu and Pizzo, 1993, J.Immunol., 150:48).α 2M and CD91 combine carboxyl terminal by α 2M partly mediate (people such as Holtet, 1994, FEBSLett., 344:242-246) and discerned Key residues (people such as Nielsen, 1996, J.Biol.Chem., 271:12909-12912).

Usually known to the Profilin enzymatic activity, α 2M combines (referring to people such as for example Hall, 1981, Biochem.Biophys.Res.Commun., 100 (1): 8-16) with the multiple protein enzyme by a plurality of binding sites.Protease and α 2M interact and cause the composite structure rearrangement, are called distortion, and it " is caught " result of the interior fracture in " bait " zone of α 2M afterwards for protease by thioesters.Conformation change exposes the required residue of receptors bind, makes α 2M-protease complex be incorporated into α 2MR.Methylamine can be induced by inductive similar conformation change of protease and fracture.The not fracture mode of the α 2M that receptor can not be discerned is commonly referred to " slowly " form (s-α 2M).The form of fracture be called " soon " form (f-α 2M) (summarize by people such as Chu, 1994, Ann.N.Y.Acad.Sci., 737:291-307).Recently shown that α 2MR can be bonded to HSP, as gp96, hsp90, hsp70 and calprotectin (people such as Basu, 2001, Immunity, 14 (3): 303-13).

Studies show that except its protease inhibitory action, when forming complex with antigen, α 2M can improve antigen and be taken in and be presented to the ability of T quadroma external by antigen-presenting cell such as macrophage, reaches as high as two orders of magnitude; (Chu and Pizzo, 1994, Lab.Invest., 71:792) and the ability of inducing T cell propagation (people such as Osada, 1987, Biochem.Biophys.Res.Commun., 146:26-31).Other evidence hint antigen and α 2M be compound have been increased by the external antibody generation of natural spleen cell (people such as Osada, 1988, Biochem.Biophys.Res.Commun., 150:883), cause experimental rabbit (people such as Chu, 1994, J.Immunol., 152:1538-1545) and mice (people such as Mitsuda, 1993, Biochem.Biophys.Res.Commun., the internal antibody in 101:1326-1331) is replied.α 2M-antigenic peptide complexes also shown inducing cytotoxic t cell response in the body (people such as Binder, 2001, J.Immunol., 166:4698-49720).

2.2. heat shock protein

Heat shock protein (HSP) is also referred to as stress protein, and is initial as being discerned by the synthetic protein of cellular response heat shock.Hsp has been divided into 5 families, Hsp100, Hsp90, Hsp70, Hsp60 and smHsp according to molecular weight.The many members that find these families subsequently can reply other stress stimulation and be induced, comprise that nutrition is deprived, in metabolism interruption, oxygen-derived free radicals and the born of the same parents or the outer pathogenic infection of born of the same parents (referring to Welch, 1993, ScientificAmerican56-64; Young, 1990, Annu.Rev.Immunol., 8:401-420; Craig, 1993, Science, 260:1902-1903; People such as Gething, 1992, Nature, 355:33-45; With people such as Lindquist, 1988, Annu.Rev.Genetics, 22:631-677).

Heat shock protein is one of existing topnotch conservative protein.For example, DnaK from the Hsp70 of escherichia coli (E.coli), has about 50% aminoacid sequence identical (people such as Bardwell, 1984, Proc.Natl.Acad.Sci., 81 with Hsp70 protein from decortication; 848-852).Hsp60 and Hsp90 family also show the level of guarding in the same high family (people such as Hickey, 1989, Mol.Cell.Biol., 9:2615-2626; Jindal, 1989, Mol.Cell.Biol., 9:2279-2283).In addition, have been found that Hsp60, Hsp70 and Hsp90 family by forming with stress protein proteins associated matter on the sequence, for example, have the aminoacid homogeneity greater than 35%, but its expression does not change in response to swashing.

Pair cell studies show that to the reaction of heat shock and other physiological stress HSP not only participates in the cytoprotective to these unfavorable conditions, and participate in stress not the necessary biochemistry of cell and immunologic process in.HSP finishes multiple chaperone function.For example, be positioned at the Hsp70 family member (people such as Lindquist of Cytoplasm, nucleus, mitochondrion or endoplasmic reticulum, 1988, Ann.Rev.Genetics, 22:631-677), participation is arrived immune cell with antigen presentation, and participates in proteinic transhipment in the normal cell, folding and assembling.HSP can conjugated protein or peptide, and discharges bonded protein or peptide in the presence of adenosine triphosphate (ATP) or low pH.

2.3.HSP-the immunogenicity of peptide complexes

People such as Srivastava proved to the inductive immunne response of the methyl cholanthrene of inbreeding mice (1988, Immunol.Today, 9:78-83).In these researchs, the molecule that discovery can be replied the different separately immunogenicity of these tumors is that the glycoprotein (gp96) and 84 of 96kDa is to the intracellular protein of 86kDa (people such as Srivastava, 1986, Proc.Natl.Acad.Sci.USA, 83:3407-3411; People such as Ullrich, 1986, Proc.Natl.Acad.Sci.USA, 83:3121-3125).With making mice to this specific tumors immunity to mouse immune, still not immune to the different tumor of antigenicity from isolating gp96 of mice or the p84/86 that suffers from specific tumors.The gene that separates and characterize coding gp96 and p84/86 shows between them and has significant homology, and show that gp96 and p84/86 are respectively the endoplasmic reticulum and the Cytoplasm counter pair (people such as Srivastava of same heat shock protein, 1988, Immunogenetics 28:205-207; People such as Srivastava, 1991, Curr.Top.Microbiol.Immunol.167:109-123).In addition, from the isolating Hsp70 of tumor to this tumor performance immunity, but to the different tumor of antigenicity do not have immunity (Udono and Srivastava, 1993, J.Exp.Med.178:1391-1396).These are observed prompting heat shock protein itself and are not had immunity, and when still forming the complex of non-covalent bond with antigenic peptides, this complex can be to the immunity of antigen peptide specific (Srivastava, 1993, Adv.Cancer Res.62:153-177; People such as Udono, 1994, J.Immunol., 152:5398-5403; People such as Suto, 1995, Science, 269:1585-1588).

Can be used for treatment and prophylaxis of cancer from the HSP of cancerous cell purification and the non-covalent bond complex of peptide, and it is open at PCT: the WO 97/10001 in WO March 20 in 96/10411,1997 on April 11st, 1996 (is respectively the United States Patent (USP) 5 of authorizing on April 12nd, 1998,750, the United States Patent (USP) 5 that on November 17th, 119 and 1998 authorized, 837,251, it all is merged in this paper as a reference) in describe to some extent.Cell separation and purification stress protein-peptide complexes from for example pathogenic infection have been described, and its can be used for treating and prevent by pathogen as in virus and other born of the same parents or the outer pathogen of born of the same parents comprise that the infection that antibacterial, protozoacide, fungus and parasite cause is (open referring to for example PCT: 21 days WO of nineteen ninety-five JIUYUE 95/24923).Also can be by making the external compound preparation immunogenicity stress protein-peptide complexes of stress protein and antigenic peptides, and this complex is used for the treatment of with the application of prophylaxis of cancer and infectious disease open at PCT: the WO 97/10000 on March 20th, the 1997 (United States Patent (USP) 6 of authorizing on February 29th, 2000,030,618) describes to some extent in.The external adoptive immunotherapy that is used for of stress protein-peptide complexes discloses the PCT that is applied in of antigen-presenting cell sensitization: the WO 97/10002 on March 20th, 1997 (also is illustrated in the United States Patent (USP) 5 of authorizing on November 16th, 1999,985,270) describe to some extent in.

2.4. α (2) macroglobulin receptor or " CD91 "

α (2) macroglobulin receptor (being called " α 2MR " or " α 2M receptor " herein interchangeably) also is known as LDL (low density lipoprotein, LDL) receptor-associated protein (" LRP ") or CD91, mainly expresses in liver, brain and Placenta Hominis.α 2M receptor is the low density lipoprotein receptor family member.The zone, extracellular of people's receptor comprises that six 50-aminoacid EGF repeat (repeat) and 31 and have the amino acid whose complement repetition of about 40-42.Complement repeats to be organized as 2,8,10 and 11 multiple bunch from the amino terminal to the carboxyl terminal, be called a bunch I, II, III and IV (people such as Herz, 1988, EMBOJ., 7:4119-4127).Research is at comprising bunch II (C1-II) that complement repeats 3-10 (CR3-10), as the main ligand binding moiety of receptor (people such as Horn, 1997, J.Biol.Chem., 272:13608-13613).α 2M receptor works in the endocytosis of multiple part.Except α 2M, other part of α 2MR comprises protein-lipid complex, lactoferrin, histiotype plasminogen activation factor (tPA), urokinase type fibrinolysin activation factor (uPA) and extracellular toxin.The other example of the part of α 2MR can find in open WO97/04794 of PCT and United States Patent (USP) 6,156,311.Therefore, α 2M receptor works in the various kinds of cell metabolic process, comprises endocytosis, antigen presentation, cholesterol regulation, contains the lipoprotein removing of ApoE and removing of Chylomicron remnants.

It is 1474 amino acid whose precursors that people α 2M is synthesized, and preceding 23 aminoacid play the effect of signal sequence, its fracture obtain 1451 amino acid whose maturation proteins (people such as Kan, 1985, Proc.Natl.Acad.Sci.U.S.A., 82:2282-2286).In the recombiant protein experiment, find 138 aminoacid (representing the amino acid/11 314-1451 of maturation protein) bind receptor of the carboxyl terminal of α 2M.This zone has been called as RBD (receptors bind zone; People such as Salvesent, 1992, FEBS Lett., 313:198-202; People such as Holtet, 1994, FEBS Lett., 344:242-246).RBD variant (RBDv), the proteolysis fragment (representing the amino acid/11 314-1451 of maturation protein) that comprises the α 2M of 15 other n terminal residues with α 2M-protease affinity bind receptor (people such as Holtet much at one, 1994, FEBS Lett., 344:242-246).

The conservative region that is present among the α macroglobulin RBD has been discerned in the sequence contrast of α 2MR part.Conserved sequence comprises the amino acid/11 366-1392 of people α 2M.Conserved residues in this zone is Phe ₁₃₆₆, Leu ₁₃₆₉, Lys ₁₃₇₀, Val ₁₃₇₃, Lys ₁₃₇₄, Glu ₁₃₇₇, Val ₁₃₈₂, Arg ₁₃₈₄(people such as Nielsen, 1996, J.Biol.Chem., 271:12909-12912).This wherein, Lys ₁₃₇₀And LYS ₁₃₇₄Show for receptors bind have crucial effects (people such as Nielsen, 1996, J.Biol.Chem., 271:12909-12912).

Part combines with α 2MR's, comprises and the combining of α 2M, and is suppressed by α 2MR associated protein (RAP).RAP be present in the 39kDa in the endoplasmic reticulum folding chaperone and for the normal process of α 2MR required.RAP can suppress combining of all α 2MR and all α 2MR parts that tried competitively.One studies show that, RAP be incorporated into complement among bunch II (C1-II) of α 2MR repeat C5-C7 (people such as Horn, 1997, J.Biol.Chem., 272:13608-13613); Another studies show that RAP is incorporated among the C1-II the multiple assembly of all giving young employees remedial-courses in general knowledge and vocational skills body weight except that C9-C10 assembly (module) (people such as Andersen, J.Biol.Chem., on March 24th, 2000, PMID:10747921; Deliver with electronic form before the printing).In RAP, discerned three domains, formed by amino acid residue 18-112,113-218 and 219-323 respectively.The part competition titrimetry in reorganization RAP territory shows, the determinant that suppresses to be tried part in the C-terminal zone of territory 1 and 3 (people such as Ellgaard, 1997, Eur.J.Biochem., 244:544-51).

Also described among the open WO 01/92474 of calendar year 2001 December disclosed PCT on the 6th CD91 as the application of heat shock protein receptor, express the cell of the CD91 that is incorporated into HSP, screening analysis, the use of regulating HSP and the interactional chemical compound of CD91 in conjunction with the antibody of CD91-HSP complex and other molecule, identification comprise the method for compositions of CD91 and be used to diagnose and treat the CD91 sequence of immune disease and proliferative disease and infectious disease.Be used for immunization therapy and the complex bonded α of antigen molecule (2) macroglobulin and be used for using this method for compositions among the disclosed WO 01/91787 description to be arranged also on 6th at PCT December of open calendar year 2001 in the diagnosis and the treatment of proliferative disease and infectious disease.People such as Binder show, the complex of the α of vitro recombination (2) macroglobulin and antigenic peptides cause specific CTL reply (people such as Binder, 2001, J.Immunol., 166:4968-4972).

2.5. antigen presentation

Main histocompatibility complex (MHC) molecule with antigen presentation to the cell surface of antigen-presenting cell.Derive from the born of the same parents still outside the born of the same parents according to antigen, it is handled by two kinds of different antigen processing approach.In the born of the same parents or endogenous protein antigen, i.e. synthetic antigen in antigen-presenting cell, by I class (MHC I) molecular presentation of MHC to the CD8+ cytotoxic T lymphocyte.On the other hand, born of the same parents outer or the synthetic antigenic determinant of external source on the cell surface of " special " or " specialty " APC (for example macrophage), be presented to the CD4+T cell (usually referring to Fundamental Immunology by the I I quasi-molecule of MHC, W.E.Paul (chief editor), NewYork:Raven Press, 1984).The differentiation of this antigen processing approach is for preventing in the immunne response process because of not adding differentiation owing to the disorganization that the release of flanking cell MHC I antigen takes place is important.

Heat shock protein has multiple chaperonin polypeptide, depend on the source of separating HSP (summary is referring to people such as Srivastava, 1998, Immunity, 8:657-665).The HSP that derives from tumor carry tumor-antigenic peptides (people such as Ishii, 1999, J.Immunology, 162:1303-1309); The gp96 goods that derive from the cell of viral infection carry virus antigen epitope (Suto and Srivastava, 1995, Science, 269:1585-1588; People such as Nieland, 1996, Proc.Natl.Acad.Sci.USA, 95:1800-1805), derive from the gp96 goods of model antigen such as ovalbumin or beta galactosidase cells transfected relevant (people such as Arnold, 1995 with epi-position accordingly, J.Exp.Med., 182:885-889; People such as Breloer, 1998, Eur.J.Immunol., 28:1016-1021).Gp96 take place with combining in vivo of peptide (Menoret and Srivastava, 1999, Biochem.Biophys.Research Commun., 262:813-818).The HSP-peptide complexes, no matter be from cell separation (people such as Tamura, 1997, Science, 278:117-120) still (people such as Blachere, 1997 of vitro recombination, J.Exp.Med., 186:1183-1406), all be excellent immunogen, and be widely used for exciting for the specific CD8+T cell response of HSP-companion's antigenic peptides.

The ability of immunne response that excites the HSP-peptide complexes depends on that peptide is to the transhipment of the MHC of antigen-presenting cell I quasi-molecule (referring to for example Suto and Srivastava, 1995, the same).The endogenous synthetic antigen of being accompanied by gp96 in the endoplasmic reticulum [ER] can contact (prime) antigenic specificity CD8+T cell (or MHC I-restriction CTL) in vivo; This contact antigen of CD8+T cell needs macrophage.Yet, do not understand the process of the gp96-peptide complexes challenging antigen specific C D8+T cell response that external source introduces fully, because there is not born of the same parents' exoantigen to translocate to definite approach that a type is presented mechanism.Yet the antigenic peptides of the born of the same parents external source relevant with HSP is caught by macrophage in some way, is directed in the endogenous approach and by MHC I molecular presentation, by the identification of CD8+ lymphocyte (Suto and Srivastava, 1995, the same; People such as Blachere, 1997, J.Exp.Med., 186:1315-22).

2.6.HSP-CD91 interact

Studies show that of people such as Basu report, heat shock protein gp96, hsp90, hsp70 and calprotectin are the additional part (people such as Basu, 2001, the same) of CD91.Gp96 embeds CD91, is arranged in to be called the segmental amino terminal fragment of p80 (people such as Binder, 2000, Nature immunology, 1:151-155; WO 01/92474).People gp96 encoding gene in advance by we chromosome 12 mapping (q24.2 q24.3) (people such as Maki, 1993, Somatic Cell Mol.Gen., 19:73-81).What is interesting is about this point, the CD91 gene mapped on the identical chromosome and be positioned at position (q13q14) little far away (people such as Hilliker, Genomics, 13:472-474).Gp96 is directly in conjunction with CD91, rather than by other part such as α 2M.In solution or the gp96 homogeneous goods that are linked to solid matrix be incorporated into CD91.In fact, the main part of CD91, α 2M suppresses rather than promotes the interaction of gp96 and CD91, proves that gp96 is the direct part of CD91.Show as 80kDa albumen in conjunction with gp96, p80, obviously be the alpha subunit of CD91 the amino terminal catabolite (people such as Binder, 2000, Nature immunology, 1:151-155).Also observe in the research formerly the CD91 of this magnitude range catabolite (people such as Jensen, 1989, Biochem.Arch., 5:171-176), and may show may be to the proteolysis cutting existence of responsive discrete extracellular domain especially among the CD91.

People's such as Basu alpha2 Macroglobulin and anti-CD91 antibody suppress to show that by the observation of presenting again of each among four kinds of HSP CD91 is unique receptor (people such as Basu, 2001, the same) of four kinds of HSP fully.Consider HSP congenital (people such as Basu, 2000, Int.Immunol., 12 (11): 1539-1546) and acquired immunity reply in conspicuous further effect, this observed result to a certain extent with counter-intuitive.Yet the data that suppress fully by two kinds of independent modes are that quite noticeable (PCT is open: 6 days WO01/92474 of calendar year 2001 December).Binder reported between hsp70 and the hsp90/gp96 with the marked difference of the competitiveness that combines the gp96 receptor (people such as Binder, 2000, J.Immunol., 165:2582-2587).Another group also observe similar difference between gp96 and the hsp70 (people such as Amold-Schild, 1999,162:3757-3760).These difference and Basu are not inconsistent at the report of the single receptor of four kinds of HSPs.They have just hinted affinity and the single acceptor interaction of different HSP to differ greatly.

Shown in people such as Binder, heat shock protein-CD91 interacts to CD91 or its fragment provide new role, i.e. the effect of pick off is not only the pick off with born of the same parents' external environment of its known in advance part based on blood plasma, and is the pick off of environment in the born of the same parents.HSP such as gp96 are molecule in the obligate born of the same parents, and it is discharged under gangrenosum acne (but not being apoptosis) cell death condition, and (PCT is open: 6 days WO of calendar year 2001 December 01/92474) in born of the same parents' external environment.Therefore, CD91 can be used as the pick off of necrosis, play the effect (people such as Savill of the receptor of the pick off of apoptotic cell death and apoptosis dead cell just as the Phosphatidylserine of scavenger receptor CD 36 and identification recently-conjugated protein, 1992, J.Clin.Invest., 90:1513-1522; People such as Fadok, 2000, Nature, 405:85-90).The interaction of macrophage and apoptotic cell causes the downward modulation (people such as Fadok of inflammatory cytokine such as TNF, 2000, ditto), and gp96-APC interacts and to cause gp96 companion peptide presenting again by the MHC I molecule of APC, T cells with antigenic specificity excitement subsequently (Suto and Srivastava, 1995, the same), and, the secretion of preceding inflammatory cytokine such as TNF, GM-CSF and IL-12.What is interesting is, α 2M, the independent part of CD91 suppresses gp96-companion's peptide presenting again by macrophage.This observed result hint gp96-of Binder accompanies presenting again in blood of peptide not take place on physiology ground, and can only be caused by the local necrosis cell death in tissue.This with blood under all test conditions in do not have gp96 or other HSP consistent fully.Generally speaking, the observation of Binder point out since the release of HSP can not cause general and fatal preceding inflammatory cytokine cascade reaction in the blood that serious tissue injury and molten born of the same parents cause may mechanism.

Therefore, might be CD91 give APC (i) by α 2M and other blood plasma part to born of the same parents' external environment sampling of blood with (ii) by HSP, the particularly gp96 family probability in the environmental sample in the born of the same parents that organize.The former allows APC to implement its primary phagocytic function, and the latter allows it to carry out its congenital and acquired immunity function.From another angle, produce the antiinflammatory signal by APC by CD36 or Phosphatidylserine identification apoptotic cell, and the interaction of APC by CD91 and non-viable non-apoptotic cell cause before the congenital and posteriority immunne response of inflammatory (referring to people such as Srivastava, 1998, Immunity, 8:657-665).

Should be with not quoting or discussing and think it is the approval of prior art of the present invention list of references herein.

3. summary of the invention

The present invention relates to use the complex treatment of α (2) macroglobulin and antigen molecule and the method and composition of prophylaxis of cancer and infectious disease, this complex derives from the patient's who suffers from cancer or infectious disease body fluid.In a preferred embodiment of the invention, α (2) macroglobulin-antigenic molecule complexes for the patient that will treat from body.The present invention part is based on applicant's following discovery: can find also therefrom to separate α (2) macroglobulin-antigenic molecule complexes to tumor or pathogen specific in the patient's who suffers from cancer or infectious disease blood flow.In addition, the applicant finds, can separate this α (2) macroglobulin-antigenic molecule complexes of q.s from patient's blood flow, is used for the autoimmune treatment.Aspect treatment of cancer, this discovery makes it possible to prepare the high degree of specificity that is oriented to the specific antigen of expressing on each independent patient's tumor, the immunotherapeutic agent of individuation, and need not at first to characterize or discern this antigen.It all is highly effective that the autoimmune treatment of this use α (2) macroglobulin complex shows as in this article in treatment for cancer and prevention.This in addition result can extend in the application for the treatment of and keeping off infection.

The applicant finds, uses the female C57BL/6 mice of α (2) macroglobulin complex immunity from the male mice purification, makes it have anti-male reaction (people such as Binder, 2002, CancerImmunity, the 2nd volume: 16).Obtain α (2) macroglobulin-male antigenic compound and be used to cause immunne response.People such as Binder prove, can be separated in the Y antigen of expressing the male mice from the serum of normal male mice, and it is the composite form with α (2) macroglobulin.People such as Binder are applied to this basic conception on the cancer immunity then, and the immunity that caused by α (2) macroglobulin-peptide complexes of vitro recombination of proof is effectively (people such as Binder, 2002, the same) in prophylaxis of tumours.

Most of tumors do not produce α (2) macroglobulin, even and those tumors that produce α (2) macroglobulin as the hepatocyte source also only in born of the same parents, produce α (2) macroglobulin.The present invention part is based on applicant's following discovery: tumor or the cell that has a pathogen are discharged into antigen in body fluid such as the blood, and they can form complex with α (2) macroglobulin there.

The invention provides treatment or prevention patient's the cancer or the method for infectious disease, described method comprises that the patient to described treatment of needs or prevention uses α (2) macroglobulin of amount of effective treatment or prevention patient's cancer or infectious disease and the complex of antigen molecule, and wherein said complex separates from the mammiferous body fluid of suffering from cancer or infectious disease.

The present invention also provides the method for treatment or prevention patient's cancer or infectious disease in addition, and it comprises step: the complex that a) separates α (2) macroglobulin and antigen molecule from the mammiferous body fluid of suffering from described cancer or infectious disease; And b) described patient is used effective treatment or prevents the described isolating complex of the amount of described cancer or infectious disease.In a specific embodiment, complex is and the colony of the complex of the bonded α of different antigen molecules (2) macroglobulin that wherein different antigen molecules comprises that described cancer or infectious disease specific antigen are had antigenic antigen molecule.In another specific embodiment, this method is used for the treatment of cancer and suffers from the patient of cancer, and antigen molecule derives from tumor.In another specific embodiments of the present invention, this method is used for the treatment of cancer and suffers from the patient of cancer, and dwindles or the number of tumors purpose reduces the effect of estimating described treatment of cancer by the patient tumors size.In another embodiment, this method is used for prophylaxis of cancer and this prevention of needs of patients, and described antigen molecule shows as described cancer specific antigen is had antigenic antigen.In another specific embodiment, this method is used to keep off infection and this prevention of needs of patients, and the specific antigen that described antigen molecule shows as described infectious disease related diseases substance has antigenic antigen.In another embodiment, this method is used for the treatment of cancer and suffers from the patient of cancer, and this method comprises the step of described patient being used chemotherapeutics before or simultaneously in step (b) in addition.In another specific embodiment, this method is used for the treatment of cancer and suffers from the patient of cancer, and this method is included in the step of induced tumor necrosis in the described mammal in addition before or simultaneously in step (a).In another specific embodiment, the step of induced tumor necrosis comprises described administration neoplasm necrosis medicine.

In another embodiment, the invention provides in the method for patient's moderate stimulation at cancer or infectious disease immunne response, it comprises: a) separate α (2) macroglobulin complex from mammiferous body fluid, and b) patient is used the isolating α (2) of institute macroglobulin complex, reply with immune stimulatory.In a specific embodiment, the patient is a human patients.In another specific embodiment, α (2) macroglobulin complex is and the colony of the complex of the bonded α of different antigen molecules (2) macroglobulin that wherein different antigen molecules comprises that described cancer or infectious disease specific antigen are had antigenic antigen molecule.In another specific embodiment, this method is used for immune stimulatory and replys patient anticancer that antagonism suffers from cancer, and this method is included in the step of induced tumor necrosis in the described mammal in addition before or simultaneously in step (a).In another specific embodiment, the step of induced tumor necrosis comprises described administration neoplasm necrosis medicine.

The present invention also provides the method for prevention mammalian cancer in addition, described method comprises α (2) macroglobulin of using effective dose and the complex of antigen molecule, described complex separates from described mammiferous body fluid, and described mammal suffers from precancerous lesion or polyp.In a specific embodiments of this method, mammal is behaved.

The present invention also provides prevention the first mammiferous method for cancer in addition, described method comprises α (2) macroglobulin of using effective dose and the complex of antigen molecule, described complex separates from the second mammiferous body fluid, and described second mammal suffers from precancerous lesion or polyp.In a specific embodiments, complex is the colony of the complex of the bonded α of different antigen molecules with one or more (2) macroglobulin, and wherein different antigen molecules comprises at least a described cancer or infectious disease specific antigen being had antigenic antigen molecule.In another specific embodiment, complex is described patient from body.In another specific embodiment, the described complex of purification before using.In another embodiment, described body fluid is the vascular fluid.In another specific embodiment, the vascular fluid is the serum that comes autoblood.In another specific embodiment, body fluid is outer ascites or cerebrospinal fluid of vascular.

In another embodiment of the invention, pharmaceutical composition is provided, it comprises that (a) is from the isolating multiple α of the mammiferous body fluid of suffering from cancer or infectious disease (2) macroglobulin-antigenic molecule complexes, described multiple complex comprises at least a following complex: it comprises α (2) macroglobulin and antigen molecule, and this antigen molecule shows as the antigenic antigen with tumor or infective agent specific antigen; (b) pharmaceutical carrier of effective dose.In a specific embodiment, body fluid is vascular fluid serum.In another specific embodiment, the vascular fluid is the serum that comes autoblood.In another specific embodiment, body fluid is outer ascites or cerebrospinal fluid of vascular.

The present invention also provides vaccine in addition, it comprises: (a) from the isolating multiple α of the mammiferous body fluid of suffering from precancerous lesion or polyp (2) macroglobulin-antigenic molecule complexes, described multiple complex comprises at least a following complex: it comprises α (2) macroglobulin and antigen molecule, and this antigen molecule shows as the antigenic antigen with tumor or infective agent specific antigen; (b) pharmaceutical carrier of effective dose.In specific embodiment, with multiple α (2) macroglobulin-antigenic molecule complexes purification.

In another embodiment of the invention, pharmaceutical composition is provided, it comprises: (a) comprise α (2) macroglobulin and have tumor or the antigenic antigen molecule of infectious disease; (b) be selected from following medicine: HSP-peptide complexes, antineoplastic agent, antibody, cytokine, antiviral agents, antifungal agent, antibiotic and chemotherapeutics; (c) pharmaceutical carrier of effective dose.In a specific embodiment, medicine is a chemotherapeutics.In another specific embodiment, with α (2) macroglobulin-antigenic molecule complexes purification.

The present invention also provides in addition increases the method that comprises the complex of α (2) macroglobulin and antigen molecule in the mammiferous body fluid, and described method is included in induced tumor necrosis in the described mammal.In a specific embodiment, the step of induced tumor necrosis comprises administration neoplasm necrosis medicine.

In another embodiment, the invention provides medicine box, it comprises α covalently or non-covalently (2) macroglobulin-antigenic molecule complexes that pharmacy can be accepted the treatment of form or prevent effective dose in one or more containers.

The present invention also provides the method for the existence that increases the complex that comprises α (2) macroglobulin and antigen molecule in the mammalian body fluid in addition, and described method is included in induced tumor necrosis in the described mammal.

In another embodiment, the invention provides preparation and comprise α (2) macroglobulin and have the method for the complex of precancerous lesion, tumor or the antigenic antigen molecule of infectious disease that it comprises: a) extract serum from the patient who suffers from precancerous lesion, tumor or infectious disease; And b) reclaims α (2) macroglobulin-antigenic molecule complexes from described serum.In a specific embodiment, this method is used for the treatment of or prophylaxis of cancer, and described method comprises the step to the complex that reclaims of described patient's administering therapeutic or prophylaxis of cancer effective dose in addition.In specific embodiment of this method, the step that reclaims α (2) macroglobulin-antigenic molecule complexes from described serum comprises: the solid phase that comprises α (2) macroglobulin binding molecule is contacted with serum enough make α (2) macroglobulin-antigenic molecule complexes be incorporated into the time of solid phase; B) remove the material that is not incorporated into described solid phase; And c) from α (2) macroglobulin-antigenic molecule complexes of solid phase elution of bound.In a specific embodiment, α (2) macroglobulin binding molecule is α (a 2) macroglobulin specific antibody.In another specific embodiment, α (2) macroglobulin binding molecule is the part binding fragment of CD91.

The present invention also provides preparation to comprise α (2) macroglobulin in addition and has the method for the complex of precancerous lesion, tumor or the antigenic antigen molecule of infectious disease, and it comprises: a) extract serum from the patient who suffers from precancerous lesion, tumor or infectious disease; And b) fractionated serum is with enriching of alpha (2) macroglobulin-antigenic molecule complexes.In a specific embodiment, the step that serum is contacted with reagent promotes the formation of covalent bond between α (2) macroglobulin and antigen molecule.In another specific embodiment, reagent is protease, ammonia, methylamine or ethamine.

In each embodiment, the body fluid of the inventive method is the vascular fluid.In other embodiments, the vascular fluid is the serum that comes autoblood.In other embodiments, body fluid is outer ascites or cerebrospinal fluid of vascular.

As used in this article, " α (2) macroglobulin " is meant α (2) macroglobulin polypeptide, and peptide binding fragment, derivant, analogies and analog.

Term " complex that comprises α (2) macroglobulin and antigen molecule " or " α (2) macroglobulin-antigenic molecule complexes " use in this article interchangeably, are meant α (2) the macroglobulin protein or polypeptide or its peptide binding fragment that are incorporated into antigen molecule.In conjunction with can be covalently or non-covalently combining between α (2) macroglobulin protein or polypeptide and the antigen molecule.

As used in this article, term " body fluid " is meant any body fluid that can be used for preparing α (2) macroglobulin-antigenic molecule complexes.Body fluid comprises vascular fluid and vascular outer fluid.The vascular outer fluid includes but not limited to ascites fluid, brain liquid, tissue lymph's liquid, colostrum and seminal fluid.In a preferred embodiment, body fluid is blood.In a concrete preferred embodiment, body fluid is the serum from blood separation.

4. accompanying drawing summary

Fig. 1 is illustrated in and uses in α (2) the macroglobulin complex mice immunized group tumor growth size (mm during 18 days ³) figure.Whether can provide preventive effect to the mouse immune that is used to first test to detect complex.A uses the PBS mice immunized as negative control, is called PBS1; B uses irradiated Meth A tumor cell mice immunized; C-F uses from 3 groups of mices (being labeled as " ascites 1-3 ") (every group of 5 mices) of the isolating α of mice serum (2) the macroglobulin-antigenic molecule complexes immunity of suffering from Intradermal Meth A tumor; G-I, 3 groups of mices (being labeled as " bleach 1-3 ") (every group of 5 mices) of using the isolating α of serum (2) the macroglobulin-antigenic molecule complexes immunity of the mice that suffers from tumor that tumor wherein handles with bleach; J uses from the isolating α of the serum of no mice with tumor (2) macroglobulin-antigenic molecule complexes mice immunized; K uses the deutero-gp96 mice immunized of MethA; L uses the compound α 2M mice immunized with MethA10 (derive from MethA tumor lysate＜peptide of 10kDa); M uses the gp96 mice immunized in liver source; N uses and the compound gp96 of MethA10 (being called gp96-MethA10) mice immunized; With 0, use PBS (being called PBS2) mice immunized.

5. detailed Description Of The Invention

The present invention relates in treatment and prophylaxis of cancer or infectious disease, use compositions and method from the isolating α of mammiferous serum (2) macroglobulin-antigenic molecule complexes.α (2) macroglobulin-antigenic molecule complexes that the present invention includes the body fluid that derives from the patient who suffers from cancer or infectious disease is used for the treatment of or prevents the method for described cancer or infectious disease.This method comprises the self-treating method of cancer and infectious disease and is used for the vaccine of prophylaxis of cancer and infectious disease.The present invention also comprises the pharmaceutical composition that contains multiple α (2) macroglobulin-antigenic molecule complexes, and pharmaceutical composition comprises and therapeutic agent α (2) macroglobulin-antigenic molecule complexes together that is used for the treatment of cancer or infectious disease.The present invention also comprises the metering method that increases α (2) macroglobulin-antigenic molecule complexes in the mammalian blood serum.

5.1. pharmaceutical composition of the present invention

The invention provides and comprise the pharmaceutical composition that can be used for treating and/or preventing cancer or infectious disease from the isolating α of mammalian blood serum (2) macroglobulin complex, this compositions also can randomly comprise the medicine that is generally used for treating cancer and infectious disease, such as but not limited to, HSP-peptide complexes, antineoplastic agent, antibody, cytokine, antiviral agents, antifungal agent, antibiotic or chemotherapeutics.Pharmaceutical composition of the present invention also can comprise and is used for the medicine described in the therapeutic alliance that directed cancer, part 5.5 be used for directed infectious disease and directed cancer of part 5.3.3 and infectious disease in the part 5.4.Pharmaceutical composition of the present invention also comprises those pharmaceutical carriers described in the part 5.9.Pharmaceutical composition of the present invention is effective especially for self-treating.This pharmaceutical composition also can comprise auxiliary agent and/or induce or increase the medicine of antigen molecule generation.In certain embodiments, medicine causes neoplasm necrosis.This paper has described the method for preparing this pharmaceutical composition.This paper has also described before separating this complex or has increased antigen molecule in the serum simultaneously and/or the method for the amount of α (2) macroglobulin-antigenic molecule complexes.

Pharmaceutical composition of the present invention and method comprise the complex of α (2) macroglobulin and antigenic peptides colony.Pharmaceutical composition shows the degeneration that can also can finally cause tumor load among the cancer patient who is treated at the inflammatory reaction at induced tumor position.Compositions by the inventive method preparation can improve individual immunocompetence and cause to the specific immunity of infective agent or to the specific immunity of pre-neoplastic cell and tumor cell.These compositionss have the morbidity that keeps off infection and the ability of development and inhibition growth of tumour cell and development.

In specific embodiment, the patient who α 2M complex is aligned the another kind treatment pattern of accepting to be used for the treatment of cancer or infectious disease uses, wherein this patient may no response for the treatment of independent treatment pattern or is had intractablely, and promptly the cancerous cell of at least some key components or pathogen are not killed or their cell division is not suppressed.Can use in the method body as known in the art or analyzed in vitro detects the effectiveness of treatment pattern.The intractable implication that this area is accepted is known in the cancer field.In one embodiment, wherein cancer or infectious disease are respectively intractable or unresponsive, and the number of cancerous cell or pathogen reduces significantly, or increases to some extent.These patients that just receiving treatment are for just accepting those of chemotherapy or radiotherapy.

In certain embodiments, compositions of the present invention comprises purified α (2) macroglobulin-antigenic molecule complexes.In these embodiments, the purity of α (2) macroglobulin-antigenic molecule complexes is at least 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100%.

5.2. the preparation of α (2) macroglobulin complex

Usually, can reclaim and purification α of the present invention (2) macroglobulin-antigenic molecule complexes from mammiferous serum by known method, comprise ammonium sulfate precipitation, acid extraction, anion or cation-exchange chromatography, cellulose phosphate (phosphocellulose) chromatography, immunoaffinity chromatography, hydroxyapatite and agglutinin chromatography.

In one embodiment, use the affinity purification technology from serum purification α (2) macroglobulin-antigenic molecule complexes.Proteinic chromatography stage division such as affinity chromatograph are as known in the art.In brief, affinity chromatograph utilizes immobilised binding partners to catch protein specifically in association reaction.The binding partners molecule of affine capture assay can comprise the antibody of α (2) macroglobulin for example or other part as specifically in conjunction with the CD91 of α (2) macroglobulin in conjunction with the territory.Perhaps, filter-binding assay use device such as solid phase surface such as filter or post are based on complex with some physics between the association reaction thing or chemical difference do not keep protein or protein complex non-specificly.Affinity chromatograph and/or filter membrane can be used for separating α (2) macroglobulin-antigenic molecule complexes from serum as herein described or other body fluid in conjunction with isolation technics.

In a specific embodiments of the present invention, separate α (2) macroglobulin-antigenic molecule complexes according to following mode from serum: make serum contact solid phase such as agarose column, it comprises the binding partners of α (2) macroglobulin, i.e. α (2) macroglobulin binding molecule.Serum is cultivated on solid phase enough made α (2) macroglobulin-antigenic molecule complexes and bonded a period of time of solid phase.Remove unconjugated material from solid phase then; And from α (2) macroglobulin-antigenic molecule complexes of solid phase elution of bound.

The binding partners of α (2) macroglobulin can be any molecule that specificity is incorporated into α (2) macroglobulin.In a preferred embodiment, α (2) macroglobulin binding molecule is the specific antibody of α (2) macroglobulin.Preferred α (2) macroglobulin specific antibody is a monoclonal antibody.In a further preferred embodiment, α (2) macroglobulin binding molecule is the part binding fragment of CD91.

Solid phase can be any surface or substrate, such as but not limited to Merlon, polystyrene, polypropylene, polyethylene, glass, nitrocellulose, dextran, nylon, polyacrylamide and agarose.Carrier structure can comprise globule; Film; Microgranule; The inner surface of reaction vessel such as microtitration plate, test tube or other reaction vessel.

In a preferred embodiment, α (2) macroglobulin-antigenic molecule complexes separates from mice serum.By using 0.04M Tris pH 7.6,0.15M NaCl, then mixture is applied to Sephacryl S300R (Sigma) post with identical buffer balance and eluting with 1: 1 dilute serum.The 65ml post is used for about 10ml serum.Detecting the positive fraction of α (2) macroglobulin and use the PD-10 post that buffer is become pH by Dot blot is 7.5 0.01M sodium phosphate buffer.In method optionally, the 0.01M sodium phosphate buffer that can in the 65ml post, use pH7.5 as buffer to save the step of exchange buffering liquid.The fraction that will contain complex is applied to concanavalin A agarose gel post.Complex with 0.2M methyl mannose pyranoside or 5% methyl mannose pyranoside elution of bound, and being used for the PD-10 post, to change buffer be the 0.05M sodium acetate buffer of pH6.0, and be applied to the equilibrated DEAE post of 0.01M sodium acetate buffer with pH6.0.With 0.13M sodium acetate buffer eluting α (2) macroglobulin is pure form, and with SDS-PAGE and immunoblotting analysis.

Above-mentioned embodiment can be used for for example comprising from mammalian cell the serum recovery and purification α (2) macroglobulin-antigenic molecule complexes of the blood of α of the present invention (2) macroglobulin-antigenic molecule complexes.During can be used for implementing, this method arrives the purification of a large amount of α (2) macroglobulin-antigenic molecule complexes.The method that does not most preferably need to reduce pH or degeneration condition is used for the purification of α (2) macroglobulin-antigenic molecule complexes.This method can be used for from eukaryotic cell such as cancerous cell, tissue, isolated cell; Or the immortal eukaryotic cell lines of or born of the same parents outer pathogenic infection interior with born of the same parents; Or derive from the patient's of pathogenic infection cell separation α (2) macroglobulin-antigenic molecule complexes.

When using above-mentioned separation and purification process, those skilled in the art can know use too harsh reagent and condition may make complex dissociation.

In another embodiment, before being used the patient, it handles complex, so that α 2M polypeptide and antigenic peptides covalency are compound from serum isolating α 2M polypeptide and antigen molecule.This paper describes the method for the α 2M-antigenic molecule complexes that forms this covalency in detail.

Usually, when α 2M mixes with protease, the fracture in " bait " zone of α 2M takes place, protease is by thioesters " seizure " and conformation change takes place, and makes α 2M complex can be incorporated into CD91.In the Proteolytic enzyme activation process of α 2M, not proteoclastic part can be covalently bonded in activatory thioesters.Not proteoclastic part also can be in using the activatory Umklapp process of heating nucleophilic be attached to (Gron and Pizzo, 1998, Biochemistry, 37 in the activatory α 2M molecule by ammonia or methylamine; 6009-6014).Can be used to prepare α 2M-antigenic compound of the present invention by this condition of the irregular seizure peptide of α 2M.This covalently bound method before describe to some extent (people such as Osada, 1987, the same; People such as Osada, 1988, the same; Chu and Pizzo, 1993, the same; People such as Chu, 1994, the same; People such as Mitsuda, 1993, the same).

For example, in a specific embodiment, in the presence of protease, ammonia or other little amine nucleophile such as methylamine and ethamine, the serum fraction of enriching of alpha 2M polypeptide and antigen molecule (but its covalent bond or not covalent bond) are mixed.The non-limitative example of spendable protease comprises that trypsin, porcine pancreatic elastase (PEP), people's neutrophilia elastoser, cathepsin G, S.aureus V-8 proteases trypsin enzyme, a-chymase, V8 protease, papain and E.C. 3.4.21.64 are (referring to people such as Ausubel, (editor), in " Current Protocolsin Molecular Biology ", Greene Publishing Associates andWiley Interscience, New York, 17.4.6-17.4.8).This paper provides α 2M polypeptide and the compound preferred illustrative scheme of antigen molecule covalency that is present in the serum that make.In a specific embodiment, following scheme can be used for increasing the quantity of α 2M-antigen molecule covalent complex.With methylamine or protease such as the 0.92mg trypsin of trypsin in about 500ml PBS (phosphate-buffered saline)) handle the serum fraction (100 μ l-5ml) comprise α 2M antigenic molecule complexes (1 μ g-20mg).Then at 37 ℃ of culture mix 5-15 minutes.If the use trypsin, then to solution add the 4mg/ml of 500ml right-amidino groups phenylmethane sulfuryl fluoride (p-APMSF) to be to suppress tryptic activity and to cultivate 2 hours at 25 ℃.Randomly, can permeate post by gelling and remove free antigen molecule.

After compound, can use mixed lymphocytes target cell for example as described below to analyze (MLTC) randomly analyzed in vitro immunogenicity α 2M-antigenic molecule complexes.In case separating immune originality complex, can before they are used, use preferred dosage regimen discussed below and excipient randomly in animal model further to its sign.

In a preferred embodiment, α (2) macroglobulin-antigenic molecule complexes separates from mice serum.By using 0.04M Tris pH 7.6,0.15M NaCl, then mixture is applied to Sephacryl S300R (Sigma) post with same buffer balance and eluting with 1: 1 dilute serum.The 65ml post is used for about 10ml serum.Detect the positive fraction of α (2) macroglobulin and use the PD-10 post buffer to be become the 0.01M sodium phosphate buffer of pH7.5 by Dot blot.In method optionally, the 0.01M sodium phosphate buffer that can in the 65ml post, use pH7.5 as buffer to save the step of exchange buffering liquid.The fraction that will contain complex is applied to concanavalin A agarose gel post.Complex with 0.2M methyl mannose pyranoside or 5% methyl mannose pyranoside elution of bound, and to be applied to the PD-10 post be the 0.05M sodium acetate buffer of pH6.0 to change buffer, and be applied to the equilibrated DEAE post of 0.01M sodium acetate buffer with pH6.0.Make α (2) macroglobulin purification with 0.13M sodium acetate buffer eluting, and by SDS-PAGE and immunoblotting analysis.Also can use other method (people such as Dubin, 1984, Biochem.International, 8 (4): 589-596 of isolated complex as known in the art; People such as Okubo, 1981, Biochem.et Biophys.Acta, 688:257-267; People such as Nieuwenhuizen, 1979, Biochem.et Biophysica.Acta, 580:129-139).

5.3. treatment is used

5.3.1. the method for treatment and prophylaxis of cancer

According to the present invention, the preferred treatment and the method for prophylaxis of cancer comprise that the individuality for the treatment of from needs separates α 2M complex and this complex is used the patient with the self-treating form.According to route of administration, use (as treatment primary carcinoma or its metastatic carcinoma) with α 2M complex separation and purification and to individuality from body, or other individualities of the cancer of needs treatments allied organization type are used, or the individuality that has the cancer risk of increase owing to family history or environmental hazard factor is used.The cancer of describing in 5.4 parts can be by pharmaceutical composition of the present invention and method treatment or prevention.

For example, can any time after the operation bring into use comprise from patient's serum separate and as above prepare the present composition of α 2M complex treat.In other embodiments, treatment can or begin in the process before operation.Yet, if the patient accepts chemotherapy, use α 2M complex behind the interval around at least usually, make immune system to recover.Therapeutic scheme can comprise that injection weekly comprises the compositions of the present invention of the α 2M complex that is dissolved in saline or other physiological compatibile solution.Each injecting pathway and position of changing, for example, it is subcutaneous to be injected at left arm for the first time, is injected at right arm for the second time, and for the third time at left abdominal part, the 4th time at right abdominal part, and the 5th time on a left side strand, and the 6th time at right strand, or the like.Identical injection site is repetition after one and a plurality of injection interval.In addition, injection is divided into two, on the same day in half of different position application dosages.Generally speaking, preceding four to six times injection is to carry out at interval with the week.Two to 50 injections subsequently serve as to carry out at interval with two weeks, serve as injection at interval with every month subsequently.

Perhaps, can use from the vaccine of the isolating α 2M of patients serum complex as prophylaxis of cancer.This vaccine can be used for by be expelled in patient's body with promote immunne response to antitumor cell, have the cell of tumor antigen.Preferably from body α 2M polypeptide-antigenic molecule complexes.Vaccine of the present invention can be before chemotherapy, in the process or use afterwards.

In a specific embodiment, cancer is a metastatic carcinoma.In another specific embodiment, cancer is a tumor.

Can monitor the immunization therapy effect of α 2M polypeptide-antigenic molecule complexes to the development of tumor disease by any method known to those skilled in the art, it includes but not limited to: a) measure delayed hypersensitivity, as the evaluation of cellular immunization; B) the lymphocytic activity of in-vitro measurements cytolysis type T-; C) level of measurement tumour specific antigen such as carcinoembryonic antigen (CEA); D) change that tomography (CT) scanning technique measures tumor form and density that uses a computer; E) use CT scan to measure the change of diameter of tumor; F) change of inferring the biomarker level of measurement particular cancers danger in the high-risk individuality; G) at the percentage ratio for the treatment of fore-and-aft survey neoplasm necrosis tissue; H) use sonogram to measure the change of tumor form; I) use nuclear magnetic resonance (MRI) to measure the change of tumor form; J) use positron emission fault imaging (PET) to measure the change of tumor form; And k) change of use ultrasonic measurement tumor form.Spendable other technology comprises scintigraphy and endoscopy.

The also preventive effect that can use α 2M polypeptide-antigenic molecule complexes carry out immunization therapy by the level evaluation of inferring biomarker that detects particular cancers danger.For example, to suffering from the dangerous individuality that increases of carcinoma of prostate, by people such as Brawer, 1992, J.Urol., people such as 147:841-845 and Catalona, 1993, JAMA, the method described in the 270:948-958 is measured serum prostate specific antigen (PSA); Or to there being the individuality of suffering from the danger of colorectum cancer to measure CEA by method as known in the art; With to the dangerous individuality that increases that suffers from breast cancer by people such as Schneider, 1982, Proc.Natl.Acad.Sci.USA, the method described in the 79:3047-3051 is measured the 16-hydroxylating of estradiol.Above-mentioned list of references is all incorporated into this paper as a reference in full.

In a specific embodiment, prevention of the present invention and treatment use be intended to increase before the operation, cancer patient's immunocompetence during operation or after the operation, with the tumour-specific immunity of inducing to cancerous cell, purpose is to suppress cancer, and final clinical purpose is the comprehensive degeneration and the elimination of cancer.

According to the present invention, compositions of the present invention comprises the complex of antigenic peptides, and described antigenic peptides derives from the cytoplasmic protein matter of cell antigen digestion and/or derives from the protein of film.α (2) macroglobulin is used the patient who suffers from cancer.In one embodiment, " treatment " be meant cancer or its at least one can discern the improvement of symptom.In another embodiment, " treatment " but be meant and the improvement of at least one measure physical parameters of related to cancer that not necessarily the patient is discernible.In another embodiment, " treatment " is meant the progress that suppresses cancer, or for example physically stablizes discernible symptom or for example stablize physical parameter to the physiology or the two exists simultaneously.

In certain embodiments, with compositions of the present invention as the antagonism this cancer preventive means to administered.As used in this article, " prevention " be meant and reduce the danger suffer from the indication cancer.In a pattern of embodiment, with compositions of the present invention as preventive means to having the administered of cancer genetic predisposition.In the another kind of pattern of embodiment, compositions of the present invention is included but not limited to the administered of chemicals and/or radiating carcinogen to contact as preventive means.

For example, in certain embodiments, with compare without the present composition, the present composition suppresses growth of tumour cell or reduces at least 99%, at least 95%, at least 90%, at least 85%, at least 80%, at least 75%, at least 70%, at least 60%, at least 50%, at least 45%, at least 40%, at least 45%, at least 35%, at least 30%, at least 25%, at least 20% or at least 10%.

Compositions by the inventive method preparation comprises the compound complex that obtains of α-2-macroglobulin and antigenic peptides colony.Compositions table reveals reducing of induced tumor size, and can finally cause by the degeneration of treatment cancer patient tumor load.Can increase the immunocompetence and the specific immunity that causes pre-neoplastic cell and tumor cell of main body by the compositions of inventive method preparation.These compositionss have the ability of prophylaxis of tumours cell advolution.

In some embodiment of the inventive method, α (2) macroglobulin-antigenic molecule complexes is used the patient with familial cancer medical history, or to being in the administered of suffering under the cancer risk owing to environmental factors.In other embodiments, use method of the present invention as preventive means treatment treatment suffer from the patient of precancerous lesion or polyp, based on the discovery of applicant to the serum isolated complex of the cancer that might form from effective antagonism.In other embodiments, use method treatment patient of the present invention, based on the discovery of the appearance that the early-stage cancer that can not survey is caused α (2) macroglobulin-antigenic molecule complexes that can be used for the prophylaxis of cancer development.In other embodiments, use method of the present invention to treat the patient who does not wherein detect cancer as yet as preventive means.

5.3.2. treatment and the method that keeps off infection

In order to treat and to keep off infection, separate α 2M-antigenic molecule complexes and be used as anti-infection vaccine from the mammiferous body fluid of suffering from infectious disease.As understood by a person skilled in the art, described herein scheme can be used for separating α 2M polypeptide-antigenic molecule complexes from any body fluid that has α (2) macroglobulin-antigenic molecule complexes.For example, cell can infected thing autoinfection.In one embodiment, can separate α 2M-antigenic molecule complexes from the antigenic serum that comprises the thing infection cell that comes from infection.In another embodiment, can from comprise derive from the born of the same parents or outside the born of the same parents the antigenic serum of pathogen separate α 2M-antigenic molecule complexes of the present invention.The invention is not restricted to treat or prevent by in the born of the same parents or the infectious disease that causes of the outer pathogen of born of the same parents.The relevant microorganism of many medical science has been described in the literature widely, for example referring to G.L.Mandell, J.E.Bennet, and R.Dolin, Mandell, Douglas, with Bennetfs Principles and Practice of Infectious Diseases, Churchill Livingstone, Philladelphia, Pennsylvania 2000, and it is incorporated into this paper as a reference in full.

Treatment or the method for optimizing that keeps off infection comprise from patient's serum separation α (2) macroglobulin complex and when needs of patients is treated isolating complex are used the patient.From the serum purification covalently or non-covalently in conjunction with the complex of the α 2M polypeptide of the antigen molecule of infective agent.

In aspect the present invention is preferred, the α 2M-antigenic molecule complexes vaccine of purification can have special purposes in treating by born of the same parents or among the human disease that pathogen causes outside the born of the same parents.Yet, should be appreciated that use the vaccine of principle described herein exploitation to can be used for treating similarly by in the born of the same parents or other mammiferous disease of causing of the outer pathogen of born of the same parents, described mammal is farming animals for example, comprising: cattle, horse, sheep, goat and pig; And house pet, comprising: cat and Canis familiaris L..

Vaccine can be prepared as the immunne response of promotion to any infectious disease pathogens described in 5.5 parts.Can monitor the immunization therapy effect of the α 2M polypeptide-antigenic molecule complexes of modification by any method known to those skilled in the art to the infectious disease development.

The complex that comprises α (2) macroglobulin and antigenic peptides colony by the compositions of inventive method preparation.Can increase the immunocompetence of main body and cause specific immunity by the compositions of the inventive method preparation infective agent.These compositionss have the ability of morbidity of keeping off infection and development.

In one embodiment, " treatment " be meant infectious disease or its at least one can discern the improvement of symptom.In another embodiment, " treatment " but be meant the improvement of at least one measure physical parameters relevant with infectious disease, but not necessarily main body is discernible.In another embodiment, " treatment " is meant the development that suppresses infectious disease, or for example physically stablely can discern symptom or for example stablize physical parameter to the physiology or both exist simultaneously.

The complex that comprises α-2-macroglobulin and antigenic peptides colony by the compositions of the inventive method preparation.Can increase the immunocompetence of main body and cause specific immunity by the compositions of the inventive method preparation infective agent.These compositionss have the ability of morbidity of keeping off infection and development.

In certain embodiments, with compositions of the present invention as the antagonism this infectious disease preventive means to administered.As used in this article, " prevention " be meant the danger that reduce to obtain the indication infectious disease.In the another kind of pattern of embodiment, compositions of the present invention is as the administered of preventive means to the infective agent of Contagious disease.

For example, in certain embodiments, with compare compositions of the present invention without the present composition, make the infective agent growth inhibited or reduce at least 99%, at least 95%, at least 90%, at least 85%, at least 80%, at least 75%, at least 70%, at least 60%, at least 50%, at least 45%, at least 40%, at least 45%, at least 35%, at least 30%, at least 25%, at least 20% or at least 10%.

5.3.3. the therapeutic alliance of treatment and prophylaxis of cancer and infectious disease

The compositions and the method for use α (2) macroglobulin complex of the present invention can be used as combined treatment with other therapeutic agent.Other therapeutic agent includes but not limited to heat shock protein-peptide complexes, antineoplastic agent, antibody, cytokine, antiviral agents, antifungal agent, antibiotic and auxiliary agent.

Therapeutic alliance is meant uses α 2M complex of the present invention with the another kind of pattern of prevention or treatment cancer and/or infectious disease.When treatment and prophylaxis of cancer, therapeutic alliance is meant uses α 2M complex of the present invention with the another kind of pattern of prevention or treatment cancer.Complex of the present invention use the effect that can strengthen cancer therapy drug, vice versa.In treatment with when keeping off infection, complex of the present invention use the effect that can strengthen anti-infectives, vice versa.Preferably, the pattern of this additional form is that non-α 2M is a pattern, and promptly this pattern does not comprise that α 2M is as component.This method is commonly referred to therapeutic alliance, auxiliary treatment or combined treatment (above-mentioned term uses in this article interchangeably).For therapeutic alliance, can be observed addition effectiveness or addition therapeutical effect.Also can expect to treat the synergistic results that effectiveness is renderd a service greater than addition.The application of therapeutic alliance also can provide than independent administering therapeutic pattern or use α 2M complex separately and better treat feature.Summation action or synergism can be turned down the dosage and/or the dosed administration frequency of any one or two kinds of patterns or avoid undesirable or disadvantageous effect.

In a plurality of specific embodiments, therapeutic alliance comprises α 2M-antigenic molecule complexes using the administered of treatment mode treatment, the treatment pattern of wherein using separately is not enough to treat main body clinically, make main body need extra effective treatment, for example, when not using α 2M complex to the unresponsive main body of treatment pattern.The method that comprises in this embodiment comprises that aligning the wherein said main body of acceptance has the administered α 2M complex of replying but experiencing the treatment pattern of side effect, recurrence, formation drug resistance etc. to treatment.This main body can or have intractablely to independent treatment pattern no response, and promptly the cancerous cell of some key component is not killed or is not suppressed their cell division or the pathogen or the infective agent of some key component are not killed at least at least.When expecting as the inventive method when using, also can improve the curative effect of treatment pattern to the method for the present invention of using the independent refractory administered α 2M of treatment pattern complex.The curative effect of treatment pattern detects can be used in the method body as known in the art or analyzed in vitro.

The intractable implication that this area is accepted is known in cancer and the infectious disease scope.In one embodiment, it is refractory or unresponsive that cancer is respectively, and wherein the number of cancerous cell does not reduce significantly, or increase to some extent.In another embodiment, it is refractory or unresponsive that infectious disease is respectively, and wherein the number of pathogen does not reduce significantly, or increase to some extent.

According to the present invention, complex of the present invention can be used for the treatment mode combinations with number of different types.Some patterns in this pattern are used in particular for the cancer or the infectious disease of particular type, and they are discussed in 5.4 and 5.5 parts respectively.Many other patterns are influential and be applicable to tumor and infectious disease usually to function of immune system.

In one embodiment, complex of the present invention is used for and one or more biological response modifier combinations, with treatment cancer or infectious disease.One class biological response modifier is a cytokine.In a this embodiment, to accepting the administered cytokine of α 2M complex.In a plurality of embodiments, can use one or more cytokines, it is selected from IL-1 α, IL-1 β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12, IFN α, IFN β, IFN γ, TNF α, TNF β, G-CSF, GM-CSF, TGF β, IL-15, IL-18, GM-CSF, INF-γ, INF-α, SLC, endothelial mononuclear cell activator protein-2 (EMAP2), MIP-3 α, MIP-3 β or mhc gene such as HLA-B7.In addition, other exemplary cytokine comprises other member of TNF family, include but not limited to TNF-α apoptosis induction ligand related (TRAIL), the relevant activation of the TNF-α inducing cell factor (TRANCE), the weak inducer (TWEAK) of TNF-α related apoptosis, CD40 part (CD40L), lymphocytotoxin α (LT-α), lymphocytotoxin β (LT-β), OX40 part (OX40L), Fas part (FasL), CD27 part (CD27L), CD30 part (CD30L), 41BB part (41BBL), APRIL, LIGHT, TL1, TNFSF16, TNFSF17 and AITR-L, or its functional part.Total summary of TNF family is referring to people such as for example Kwon, 1999, Curr.Opin.Immunol., 11:340-345.Preferably, before the treatment pattern, use α 2M complex.In a specific embodiment, complex of the present invention is applied to the patient who just accepts with cyclophosphamide and IL-12 treatment of cancer with combinations.

In another embodiment, complex of the present invention is used for and one or more biological response modifier combinations, and described biological response modifier is the agonist or the antagonist of multiple part, receptor and signal transducers.For example, biological response modifier includes but not limited to toll sample (Toll-like) receptor (TLR-2, TLR-7, TLR-8 and TLR-9); LPS; The agonist of 41BB, OX40, ICOS and CD40; The antagonist of Fas part, PD1 and CTLA-4.These agonist and antagonist can be antibody, antibody fragment, peptide, plan peptide compounds and polysaccharide.

In another embodiment, complex of the present invention is used for one or more biological response modifier combinations as immunostimulatory nucleic acid.Many in these nucleic acid are to comprise the not oligonucleotide of methylated CpG motif, promote vertebrates lymphocyte mitosis, and known enhance immunity are replied.Referring to people such as Woolridge, 1997, Blood, 89:2994-2998.This oligonucleotide is described in international monopoly open WO 01/22972, WO 01/51083, WO98/40100 and WO 99/61056 to some extent, and described open all full text is incorporated this paper into as a reference, and in U.S. Patent Publication 6,207,646,6,194,388,6,218,371,6,239,116,6,429,199 and 6, describe to some extent in 406,705, described open all full text is incorporated this paper into as a reference.The immunostimulating oligonucleotide of other kind as the sulfo-phosphide oligodeoxyribonucleotide that comprises YpG-motif and CpR-motif by people such as Kandimalla at " Effect of Chemical Modifications of cytosine and Guanine ina CpG-Motif of Oligonucleotides:Structure-ImmunostimulatoryActivity Relationships. ", Bioorganic﹠amp; Describe among the Medicinal Chemistry, 9:807-813 (2001), it incorporates this paper into as a reference in full.Also comprise the immunostimulating oligonucleotide that does not contain the CpG dinucleotide, it strengthens antibody response when using (comprise low dose use) by mucosal route or using by the parenteral route high dose, often the same with the effect of CpG nucleic acid, yet, this reply into Th2-tendency partially (IgG1＞＞IgG2a).Referring to U.S. Patent Publication 20010044416 A1, it incorporates this paper into as a reference in full.Can detect the active method of immunostimulating oligonucleotide with above-mentioned patent with openly.In addition, the immunostimulating oligonucleotide can be connected at phosphate backbone, sugar, nucleotide base and nucleotide closes interior modification, to regulate activity.This be modified to well known by persons skilled in the art.

In another embodiment, complex of the present invention is used for and one or more auxiliary agent combinations.Auxiliary agent can be used separately or mix with complex of the present invention and be present in the compositions.The auxiliary agent of general auxiliary agent for can non-intestinal using.The general auxiliary agent comprises producing stores auxiliary agent, the auxiliary agent of stimulating immune system and the auxiliary agent useful to the both that (depot) acts on.As used herein, thus the auxiliary agent that produces the storage effect is antigen slowly to be discharged in vivo prolong the auxiliary agent of immunocyte to the antigen contact.This analog assistant includes but not limited to Alumen (as aluminium hydroxide, aluminum phosphate); Or emulsion is preparation, comprise mineral oil, non-mineral oil, Water-In-Oil or oil-water-fat liquor, oil in water emulsion such as Montanide auxiliary agent Seppic ISA series (as Montanide ISA720, AirLiquide, Paris, France); MF-59 (with sorbester p37 and the stable water cornerite zamene emulsion of Tween 80, Chiron Corporation, Emeryville, Calif.); And PROVAX (comprise the O/w emulsion of stablizing detergent and micelle plasticizer, IDEC, Pharmaceuticals Corporation, San Diego, Calif.).

Other auxiliary agent stimulating immune system for example makes immunocyte produce justacrine cytokine or IgG.This analog assistant includes but not limited to immunostimulatory nucleic acid, as the CpG oligonucleotide; From the Saponin of the bark purification of Q. Saponaria officinalis tree, as QS21; Poly-[two (carboxylation phenoxy group) phosphonitrile] (PCPP polymer; Virus Research Institute, USA); The derivant of lipopolysaccharide (LPS) such as monophosphoryl lipid A (MPL, Ribi ImmunoChem Research, Inc., Hamilton, Mont.), muramyldipeptide (MDP, Ribi) and threonyl-muramyldipeptide (t-MDP, Ribi); OM-174 (the glycosamine disaccharide relevant with lipid A, OM Pharma SA, Meyrin, Switzerland); With the leishmania elongation factor (the leishmania protein of purification, Corixa Corporation, Seattle, Wash.).In a preferred embodiment, complex of the present invention is used for making up such as but not limited to the CG oligonucleotide with QS21 and another kind of immunostimulatory nucleic acid.

Other general auxiliary agent is for producing the auxiliary agent of storage effect and stimulating immune system.These chemical compounds are the chemical compound that has both the recognition function of above-mentioned general auxiliary agent.This analog assistant include but not limited to ISCOM (the immunostimulating complex, it comprises and mixes Saponin, lipid and form the granule with the virus size that can hold antigenic hole, CSL, Melbourne, Australia); SB-AS2 (SmithKline Beecham adjuvant systems #2, it is the O/w emulsion that comprises MPL and QS21, SmithKline Beecham Biologicals[SBB], Rixensart, Belgium); SB-AS4 (SmithKline Beecham adjuvant systems #4, it comprises Alumen and MPL, SBB, Belgium); The nonionic block copolymer of formation micelle such as CRL 1005 (these comprise the straight chain hydrophobicity polypropylene oxide by polyethylene glycol oxide chain formation side chain, Vaxcel, and Inc., Norcross, Ga.); With Syntex Adjuvant Formulation (SAF comprises the O/w emulsion of Tween 80 and nonionic block copolymer, Syntex Chemicals, Inc., Boulder, Colo.).

Spendable mucosa auxiliary agent is for inducing the auxiliary agent of the mucosal immune response of main body when being applied to mucomembranous surface with complex of the present invention according to the present invention.The mucosa auxiliary agent includes but not limited to CpG nucleic acid (as the disclosed patent application WO 99/61056 of PCT), bacteriotoxin: include but not limited to CTB subunit (CTB) (people such as Wu, 1998 as cholera toxin (CT), CT derivant; People such as Tochikubo, 1998); CTD53 (Val is to Asp) (people such as Fontana, 1995); CTK97 (Val is to Lys) (people such as Fontana, 1995); CTK104 (Tyr is to Lys) (people such as Fontana, 1995); CTD 53/K63 (Val is to Asp, and Ser is to Lys) (people such as Fontana, 1995); CTH54 (Arg is to His) (people such as Fontana, 1995); CTN107 (His is to Asn) (people such as Fontana, 1995); CTE114 (Ser is to Glu) (people such as Fontana, 1995); CTE112K (Glu is to Lys) (people such as Yamamoto, 1997a); CTS61F (Ser is to Phe) (people such as Yamamoto, 1997a, 1997b); CTS106 (Pro is to Lys) (people such as Douce, 1997, people such as Fontana, 1995); And CTK63 (Ser is to Lys) (people such as Douce, 1997, people such as Fontana, 1995), little band links toxin (Zonula occludentstoxin), zot, heat-labile enterotoxin of E, coli, labile toxin (LT), LT derivant and includes but not limited to LTB subunit (LTB) people such as (, 1998) Verweij; LT7K (Arg is to Lys) (people such as Komase, 1998, people such as Douce, 1995); LT61F (Ser is to Phe) (people such as Komase, 1998); LT112K (Glu is to Lys) (people such as Komase, 1998); LT118E (Gly is to Glu) (people such as Komase, 1998); LT146E (Arg is to Glu) (people such as Komase, 1998); LT192G (Arg is to Gly) (people such as Komase, 1998); LTK63 (Ser is to Lys) (people such as Marchetti, 1998, people such as Douce, 1997,1998, people such as DiTommaso, 1996); And LTR72 (Ala is to Arg) (people such as Giuliani, 1998), pertussis toxin, PT, PT. (people such as Lycke, 1992, Spangler BD, 1992, Freytag and Clemments, 1999, people such as Roberts, 1995, people such as Wilson, 1995) comprise PT-9K/129G (people such as Roberts, 1995, people such as Cropley, 1995); Toxin derivant (seeing following) (people such as Holmgren, 1993, people such as Verweij, 1998, people such as Rappuoli, 1995, Freytag and Clements, 1999); The lipid A derivant (as single phosphono lipid A, MPL) (people such as Sasaki, 1998, people such as Vancott, 1998; Muramyldipeptide (MDP) derivant (people such as Fukushima, 1996, people such as Ogawa, 1989, people such as Michalek, 1983, people such as Morisaki, 1983); Bacterial outer membrane protein (as outer surface A albumen (OspA) lipoprotein of Borrelia burgdoyferi bacterium (Borrelia Burgdorferi), the outer membrane protein of Neisseria meningitidis (Neisseria meningitidis)) (people such as Marinaro, 1999, people such as Van de Verg, 1996); Oil in water emulsion (as MF59) (people such as Barchfield, 1999, people such as Verschoor, 1999, O ' Hagan, 1998); Aluminum salt (people such as Isaka, 1998,1999); And Saponin (as QS21) Aquila Biopharmaceuticals, Inc., Worster, Me.) (people such as Sasaki, 1998, people such as MacNeal, 1998), and ISCOMs, MF-59 is (with sorbester p37 and the stable water cornerite zamene emulsion of Tween 80; ChironCorporation, Emeryville, Calif.); The Seppic ISA series of Montanide auxiliary agent is (as Montanide ISA 720; AirLiquide, Paris, France); PROVAX (comprises the O/w emulsion that stable detergent and micelle are composed plasticizer; IDECPharmaceuticals Corporation, San Diego, Calif.); SyntextAdjuvant Formulation (SAF; Syntex Chemicals, Inc., Boulder, Colo.); Poly-[two (carboxylated phenoxy group) phosphonitrile] (PCPP polymer; Virus ResearchInstitute, USA) and the leishmania elongation factor (Corixa Corporation, Seattle, Wash.).

In another embodiment, complex of the present invention and one or more immunotherapy medicaments such as antibody and vaccine combine and use.In a preferred embodiment, antibody has the interior therapeutic and/or the preventive effect of antagonism cancer and/or infectious disease.The example of treatment and prevention antibody include but not limited to MDX-010 (Medarex, NJ), its for the humanization that is used for the clinical treatment carcinoma of prostate at present anti--CTLA-4 antibody; SYNAGIS  (Medlmmune, MD), it is humanization anti respiratory syncytial virus (RSV) monoclonal antibody that is used for the treatment of the rsv infection patient; (Genentech, CA), it is the Humanized anti-HER 2 monoclonal antibody that is used for the treatment of the metastatic breast cancer patient to HERCEPTIN  (Trastuzumab).Other example is that humanization resists-CD18F (ab ') ₂(Genentech); CDP860, it is the anti-CD18F of humanization (ab ') ₂(Celltech, UK); PR0542, it is the anti-HIV gp120 antibody (Progenics/Genzyme Transgenics) that merges with CD4; Ostavir, its behaviour anti-hepatitis B virus antibody (Protein Design Lab/Novartis); PROTOVIR ^TM, it is the anti-CMV IgG1 of humanization antibody (Protein Design Lab/Novartis); MAK-195 (SEGARD), it is mouse-anti TNF-α F (ab ') ₂(Knoll Pharma/BASF); IC14, it is anti-CD 14 antibody (ICOS Pharm); The anti-VEGF IgG1 of humanization antibody (Genentech); OVAREX ^TM, it is a Mus anti-CA 125 antibody (Altarex); PANOREX ^TM, it is a mouse-anti 17-IA cell surface antigen IgG2a antibody (GlaxoWellcome/Centocor); BEC2, it is mouse-anti idiotype (GD3 epi-position) IgG antibody (ImClone System); IMC-C225, it is chimeric anti-EGFR IgG antibody (ImCloneSystem); VITAXIN ^TM, it resists-α V β 3 alpha 2 integrin antibodies (AppliedMolecular Evolution/MedImmune) for humanization; Campath 1H/LDP-03, it is a humanized anti-CD 52 IgG1 antibody (Leukosite); Smart M195, it is humanization anti-CD 33 IgG antibody (Protein Design Lab/Kanebo); RITUXAN ^TM, its be chimeric anti-CD20 IgG1 antibody (IDEC Pharm/Genentech, Roche/Zettyaku); LYMPHOCIDE ^TM, it is the anti-CD22 IgG of a humanization antibody (Immunomedics); Smart ID10, it is the anti-hla antibody of humanization (Protein Design Lab); ONCOLYM ^TM(Lym-1), be radiolabeled mouse-anti HLA DIAGNOSTIC REAGENT antibody (Techniclone); ABX-IL8 is the anti-IL8 antibody of people (Abgenix); Anti-CD11a is humanization IgG1 antibody (Genentech/Xoma); ICM3 is the anti-ICAM3 antibody of humanization (ICOS Pharm); IDEC-114 is the long anti-CD80 antibody of sourceization (IDECPharm/Mitsubishi) of spirit; ZEVALIN ^TM, be radiolabeled mouse-anti CD20 antibody (IDEC/Schering AG); IDEC-131 is humanization anti-CD40L antibodies (IDEC/Eisai); IDEC-151 is the long source anti-CD 4 antibodies (IDEC) of spirit; IDEC-152 is the long anti-CD23 antibody of sourceization (IDEC/Seikagaku) of spirit; The anti-CD3 of SMART is the anti-CD3IgG of humanization (Protein Design Lab); 5G1.1 be the humanization anticomplementary factor 5 (C5) antibody (Alexion Pharm); D2E7 is the anti-TNF-Alpha antibodies of humanization (CAT/BASF); CDP870 is humanization anti-TNF-α Fab fragment (Celltech); IDEC-151 is the long anti-CD4IgG1 antibody of sourceization (IDEC Pharm/SmithKline Beecham) of spirit; MDX-CD4 is the anti-CD4 IgG of a people antibody (Medarex/Eisai/Genmab); CDP571 is the anti-TNF-α of a humanization IgG4 antibody (Celltech); LDP-02 is humanized anti-alpha 4 β, 7 antibody (LeukoSite/Genentech); OrthoClone OKT4A is the anti-CD4 IgG of a humanization antibody (Ortho Biotech); ANTOVA ^TM, be humanization anti-CD 40 L IgG antibody (Biogen); ANTEGREN ^TMBe the anti-VLA-4 IgG of humanization antibody (Elan); MDX-33 is people's anti-CD 64 (Fc γ R) antibody (Medarex/Centeon); SCH55700 is the anti-IL-5 IgG4 of a humanization antibody (Celltech/Schering); SB-240563 and SB-240683 are respectively anti-IL-5 of humanization and IL-4 antibody (SmithKline Beecham); RhuMab-E25 is the anti-IgE IgG1 of a humanization antibody (Genentech/Norvartis/Tanox Biosystems); ABX-CBL is a mouse-anti CD-147 IgM antibody (Abgenix); BTI-322 is a rat anti CD2 IgG antibody (Medimmune/Bio Transplant); Orthoclone/OKT3 is a mouse-anti CD3 IgG2a antibody (ortho Biotech); SIMULECT ^TMBe chimeric anti-CD25 IgG1 antibody (Novartis Pharm); LDP-01 is the anti-β of humanization ₂-integrin IgG antibody (LeukoSite); Anti-LFA-1 is mouse-anti CD18F (ab ') ₂(Pasteur-Merieux/Immunotech); CAT-152 is people's anti-TGF-beta ₂Antibody (Cambridge AbTech); With Corsevin M be chimeric anti-factor VII antibody (Centocor).In a preferred embodiment, complex of the present invention and anti-CTLA 4 antibody combined administration.In a further preferred embodiment, complex of the present invention and anti-41BB antibody combined administration.The above-mentioned immunocompetence drug of enumerating and any other immunocompetence drug can comprise that the scheme of immunocompetence drug supplier recommendation is used according to any scheme well known by persons skilled in the art.

In treatment or when keeping off infection, with α (2) macroglobulin-antigenic molecule complexes to because environment or familial factor are in the patient who suffers under the infectious disease danger uses.In other embodiments, based on using the method for the present invention patient that treatment has the infectious disease early symptom as preventive means from effective idea of resisting the serum isolated complex of possibility development infectious diseases.In other embodiments, may cause the conviction of appearance of α (2) macroglobulin-antigenic molecule complexes of the development that can be used for keeping off infection based on undetectable pathogen early infection, use method treatment of the present invention not have the patient of infectious disease symptom.In other embodiments, may suffer from the idea of nd infectious disease still, use method of the present invention as the healthy patient of preventive means treatment based on the patient.

For determining the effectiveness of the compositions and methods of the invention, the various standard analytical processes that those skilled in the art can use detection to infect include but not limited to the antigen titration analysis, use round pcr to cultivate as detecting viral specific nucleic acids measurement viral load, blood cell analysis, plaque detection, phage analysis or bacteria samples.This method that is used to detect and measure infection level is well known to a person skilled in the art.

In another embodiment, in the situation of treatment for cancer and prevention method, each said method comprises the administered α 2M complex of the combination that aligns the treatment for cancer pattern of receiving treatment, and preferably uses the α 2M complex of purification.Preferably, α 2M complex is incorporated into and shows the cancer types that aligns treatment antigenic antigen molecule is arranged.

5.4. target cancer

In one embodiment, method of the present invention and therapeutic alliance thereof comprise uses compositions of the present invention, compositions includes one or more patterns that help prevent or treat the assistance application of cancer, and pattern includes but not limited to chemotherapeutics, immunotherapeutic agent, anti-angiogenic agent, cytokine, hormone, antibody, polynucleotide, radiation and photodynamic therapy agent.In specific embodiment, relate to the therapeutic alliance of using pharmaceutical composition of the present invention and can be used for the prophylaxis of cancer recurrence, suppress cancer metastasis or suppress cancer or metastatic cancer growth and/or diffusion.

Can include but not limited to people's sarcoma and cancer by the cancer types of method of the present invention and medicine composite for curing or prevention, as fibrosarcoma; Myxosarcoma; Liposarcoma; Chondrosarcoma; Osteogenic sarcoma; Chordoma; Angiosarcoma; Endotheliosarcoma; Lymphangiosarcoma; Lymphangioendothelial sarcoma (lymphangioendothelio sarcoma); Synovioma; Mesothelioma; Ewing's sarcoma; Leiomyosarcoma; Rhabdomyosarcoma; Colon cancer; Cancer of pancreas; Breast carcinoma; Ovarian cancer; Carcinoma of prostate; Squamous cell carcinoma; Basal cell carcinoma; Adenocarcinoma; Syringocarcinoma; Sebaceous gland carcinoma; Papillary carcinoma; Papillary adenocarcinoma; Cystadenocarcinoma; Medullary carcinoma; Bronchogenic carcinoma; Renal cell carcinoma; Hepatoma; Cancer of biliary duct; Choriocarcinoma; Spermocytoma; Embryonal carcinoma; Wilms' tumor (wilms ' tumor); Cervical cancer; Carcinoma of testis; Pulmonary carcinoma; Small cell lung cancer; Bladder cancer; Epithelial cancer; Glioma; Astrocytoma; Medulloblastoma; Craniopharyngioma; Ependymoma; Pinealoma; Hemangioblastoma; Acoustic neuroma; Oligodendroglioma; Meningioma; Melanoma; Neuroblastoma; Retinoblastoma; Leukemia such as acute lymphoblastic leukemia and acute myeloid leukemia (myeloblastosis, promyelocytic leukemia, bone marrow mononuclear cell leukemia, monocytic leukemia and erythroleukemia); Chronic leukemia (chronic myelocytic (granulocytic) leukemia and chronic lymphocytic leukemia); With polycythemia vera, lymphoma (Hokdkin disease and Fei Hejiejinshi disease), multiple myeloma, Waldenstr  m ' s macroglobulinemia and heavy chain disease.

Immunization therapy provided by the invention is that the required multiple reason of cancer patient has, the first, if the cancer patient is subjected to immunosuppressant, then anesthesia surgery and chemotherapy subsequently may make immunosuppressant worsen.Can prevent or reverse this immunosuppressant in the suitable immunization therapy of surgical operation between early stage.This may make and infect complication still less and accelerating wound healing.The second, operation back gross tumor volume is minimum, and therefore immunization therapy is effective probably in this case.The 3rd, tumor cell may be discharged in the circulation when operation, at this moment uses effective immunization therapy and can remove these cells.

Prevention of the present invention and Therapeutic Method are at the immunocompetence that increases the cancer patient before operation, after when operation or the operation, and induce tumour-specific immunity at cancerous cell, its objective is the inhibition cancer, its final clinical purpose is the comprehensive degeneration and the elimination of cancer.Method of the present invention and pharmaceutical composition for example also can be used for being in the dangerous individuality that increases of specific types of cancer owing to family history or environmental hazard factor.

In a plurality of embodiments, except complex of the present invention, use one or more cancer therapy drugs and treat the cancer patient.Cancer therapy drug is meant any molecule or the chemical compound that helps tumor or treatment of cancer.The example that is used for the cancer therapy drug of method of the present invention includes but not limited to: acivicin; Aklavine; The hydrochloric acid acodazole; Acronine; Adozelesin; IL-2; The first pyrimidine; Ambomycin; Acetic acid A Mei anthracene; Aminoglutethimide; Amsacrine; Anastrozole; Antramycin; Asparaginase; Asperlin; Azacitidine; Azatepa; Azotomycin; Batimastat; Benzodepa; Bicalutamide; Bisantrene hydrochloride; Bisnafide two methanesulfonates; Bizelesin; Bleomycin Sulphate; Brequinar sodium; Bropirimine; Busulfan; Actinomycin C; Calusterone; Caracemide; Carbetimer; Carboplatin; Carmustine; Carubicin hydrochloride; Carzelesin; Cedefingol; Chlorambucil; Cirolemycin; Cisplatin; Cladribine; The methanesulfonic acid crisnatol; Cyclophosphamide; Cytosine arabinoside; Dacarbazine; Dactinomycin; Daunorubicin hydrochloride; Decitabine; Dexormaplatin; Dezaguanine; The methanesulfonic acid Dezaguanine; Diaziquone; Docetaxel; Amycin; Doxorubicin hydrochloride; Droloxifene; The citric acid droloxifene; Dromostanolone propionate; Diazomycin; Edatrexate; Eflornithine hydrochloride; Elsamitrucin; Enloplatin; Enpromate; Epipropidine; Epirubicin hydrochloride; Erbulozole; Esorubicin hydrochloride; Estramustine; Estramustine phosphate sodium; Etanidazole; Etoposide; The phosphoric acid etoposide; Etoprine; CGS-16949A; Fazarabine; Fenretinide; The 5-fluorouracil deoxynucleoside; Fludarabine phosphate; Fluorouracil; Flurocitabine; Fosquidone; Fostriecin sodium; Gemcitabine; Gemcitabine hydrochloride; Hydroxyurea; Idarubicin hydrochloride; Ifosfamide; Ilmofosine; Interleukin I I (comprising recombinant interleukin II or rIL2); Intederon Alpha-2a; Interferon Alpha-2b; Interferon alfa-n1; Alferon N; Interferon beta-Ia; Interferon gamma-Ib; Iproplatin; Irinotecan hydrochloride; Somatuline Acetate; Letrozole; Leuprorelin acetate; Liarozole hydrochloride; Lometrexol sodium; Luo Mositing; Losoxantrone hydrochloride; Masoprocol; Maytansine; Mustine hydrochlcride; Megestrol acetate; Melengestrol acetate; Melphalan; Menogaril; Mercaptopurine; Methotrexate; Methotrexate sodium; Metoprine; Meturedepa; Mitindomide; Mitocarcin; Mitochromine mitocromine B-35251; Mitogillin; Mitomalcin; Mitomycin; Mitosper; Mitotane; Mitoxantrone hydrochloride; Mycophenolic acid; Nocodazole; Nogalamycin; Ormaplatin; Oxisuran; Paclitaxel; Pegaspargase; Peliomycin; Pentamustine; Peplomycin sulfate; Perfosfamide; Pipobroman; Piposulfan; The hydrochloric acid piroxantrone; Plicamycin; Plomestane; Porfimer sodium; Porfiromycin; Prednimustine; Procarbazine hydrochloride; Puromycin; Puromycin hydrochloride; Pyrazofurin; Riboprine; Rogletimide; Safingol; The hydrochloric acid Safingol; Semustine; Simtrazene; Sparfosate Sodium; Sparsomycin; Spirogermanium hydrochloride; Spiromustine; Spiroplatin; Rufocromomycin; Streptozotocin; Sulofenur; Talisomycin; Tecogalan sodium; Ftorafur; Teloxandrone hydrochloride; Temoporfin; Teniposide; Teroxirone; Testolactone; ITG; 2-amino-Ismipur; Plug is for group; Thiazole furan quinoline; Tirapazamine; Toremifene Citrate; Trestolone acetate; The phosphoric acid triciribine; Trimetrexate; The glucuronic acid trimetrexate; Triptorelin; Tubulozole hydrochloride; Uracil mustard; Uredepa; Vapreotide; Verteporfin; Vinblastine sulfate; Vincristine sulfate; Vindesine; Vindesine sulfate; The sulphuric acid vinepidine; The sulphuric acid vinglycinate; The sulphuric acid vinleurosine; Vinorelbine tartrate; The sulphuric acid vinrosidine; The sulphuric acid vinzolidine; R 83842; Zeniplatin; Neocarzinostain NCS; Zorubicin hydrochloride.

Spendable other cancer therapy drug includes but not limited to: 20-table-1,25-dihydroxy vitamin d3; 5-ethinyluracil; Abiraterone; Aklavine; Acylfulvene; Adecypenol; Adozelesin; Interleukin-2; The ALL-TK antagonist; Hex A; Ambamustine; Amidox; Amifostine; Aminolevulinic acid; Amrubicin; Amsacrine; Anagrelide; Anastrozole; Andrographolide; Angiogenesis inhibitor; Antagonist D; Antagonist G; Antarelix; Anti-dorsalization BMPs-1; Antiandrogen carcinoma of prostate (prostatic carcinoma); Estrogen antagonist; Antitumor forms; Antisense oligonucleotide; Glycine Ai Fei ground can be peaceful; The apoptogene regulator; The apoptosis controlling agent; Apurinic acid; Ara-CDP-DL-PTBA; The arginine deaminase; Asulacrine; Atamestane; Atrimustine; A Xinasiting (axinastatin) 1; A Xinasiting 2; A Xinasiting 3; Azasetron; Azatoxin; Azatyrosine; Baccatin III derivative; Balanol; Batimastat; The BCR/ABL antagonist; Benzochlorins; Benzoylstaurosporine; The beta-lactam derivant; β-alethine; β clarithromycin B; Belulinic acid Betulinic acid; The bFGF inhibitor; Bicalutamide; Bisantrene; Bisaziridinylspermine; Bisnafide; Bistratene A; Bizelesin; Breflate; Bropirimine; Budotitane; Butyl thionine imines; Calcipotriol; Calphotin (calphostin C); Camptothecin derivative; Canary pox IL-2; Capecitabine; Methanamide-amino-triazole; The carboxyamino imidazoles; CaRest M3; CARN 700; The deutero-inhibitor of cartilage; Carzelesin; Casein kinase 2 enzyme inhibitor (ICOS); Castanospermine; Cecropin B; Cetrorelix; Chlorlns; Chloro-quinoxaline; Sulphanilamide; Cicaprost; Along porphyrin; Cladribine; The clomifene analog; Clotrimazole; Courlene mycin A; Courlene mycin B; Combretastatin A4; The combretastatin analog; Conagenin; Mediterranean sponge 816; Crisnatol; Latent algin (cryptophycin) 8; Latent algin A derivant; Coumarin A; Encircle penta anthraquinone; Cycloplatam; Cypemycin; Cytosine arabinoside ocfosfate; Cytolytic factor; Hexestryl diphosphate; Dazopride; Decitabine; The dehydrogenation didemnun B; Deslorelin; Dexamethasone; Right ifosfamide; Dexrazoxane; Dexverapamil; Diaziquone; Didemnin B; Didox; Diethyl norspermine; Dihydro-5-azepine cytosine; 9-dihydro paclitaxel; Dioxamycin; The diphenyl spiromustine; Docetaxel; Tadenan; Dolasetron (dolasetron); Doxifluridine; Droloxifene; Dronabinol; Duocarmycin SA; Ebselen; Ecomustine; Edelfosine; Edrecolomab; Eflornithine; Elemene; Emitefur; Epirubicin; Epristeride; The estramustine analog; Estrogen agonist; Estrogen antagonist; Etanidazole; The phosphoric acid etoposide; Exemestane; Fadrozole; Fazarabine; Fenretinide; Filgrastim; Fei Nasi carries (finasteride); Flavopiridol; Flezelastine; Fluasterone; Fludarabine; Hydrochloric acid fluorodaunorunicin; Forfenimex; Formestane; Fostriecin; Fotemustine; The texaphyrin gadolinium; Ganite (Fujisawa).; Galocitabine; Ganirelix; The gelatinase inhibitor; Gemcitabine; The glutathion inhibitor; Hepsulfam; Heregulin; Hexamethylene bisacetamide; Hypericin; Ibandronic acid; Idarubicin; Idoxifene; Idramantone; Ilmofosine; Ilomastat; The imidazoles acridone; Imiquimod; The immunostimulating peptide; The type-1 insulin like growth factor acceptor inhibitor; The interferon agonist; Interferon; Interleukin; Iobenguane; Iododoxorubicin; The 4-ipomeanol; Iroplact; Irsogladine; The Yi Benjia azoles; Isohomohalicondrin B; Itasetron; Jasplakinolide; Kahalalide F; Sheet spiral shell element-N triacetate; The slow release lanreotide; Leinamycin; Lenograstim; The sulphuric acid lentinan; Leptolstatin; Letrozole; The hypercytosis inhibitive factor; The leukocyte interferon-alpha; Leuprorelin+estrogen+Progesterone; Leuprorelin; Levamisole; Liarozole; Linear polyamine analog; Lipotropy two glycopeptides; The lipotropy platinum compounds; Lissoclinamide 7; Lobaplatin; Lombricine (lombricine); Lometrexol; Lonidamine; Losoxantrone; Lovastatin; The Luo Suoli shore; Lurtotecan; Lutetium texaphyrin; Lysofylline; Polypeptide dissociates; Maitansine; Mannostatin A; Marimastat; Masoprocol; Maspin; Substrate lysin inhibitor; Matrix metallo-proteinase inhibitor; Menogaril; Mei Balong; U.S. Tequ woods (meterelin); Methioninase; Metoclopramide; The MIF inhibitor; Mifepristone; Mi Tefuxin; Mirimostim; Mismatching double stranded; Mitoguazone; Mitolactol; Mitomycin analogs; Mitonafide; Mitotoxin fibroblast growth factor saporin (saporin); Mitoxantrone; Mofarotene; Molgramostim; Monoclonal antibody, human chorionic gonadotropin; Monophosphoryl lipid matter A+myobacterium cell wall sk; Moperone; Many drug resistant genes inhibitor; Treatment based on many tumor inhibitors 1-; The nitrogen mustards anticarcinogen; Indian Ocean sponge B; The mycobacteria cell wall extracts; Myriaporone; The N-Tacedinaline; N-substituted benzene formyl amine; The naphthalene Rayleigh; Nagrestipen (nagrestip); Allylnoroxymorphone+pentazocine; Napavin; Naphterpin; Nalbuphine; Naphthalene reaches platinum; Nemorubicin; Neridronic acid; Neutral endopeptidase; Nilutamide; Nisamycin; The nitric oxide regulator; The nitrous oxide antioxidant; Nitrullyn; 06-benzyl guanidine; Octreotide; Okicenone; Oligonucleotide; Onapristone; Ondansetron; Ondansetron oracin; Oral cytokine induction agent; Ormaplatin; Osaterone; Oxaliplatin; Oxaunomycin; Paclitaxel; Paclitaxel analogs; Paclitaxel derivant; Palauamine; Palmitoylrhizoxin; Pamidronic acid; The panaxatriol; Panomifene; Secondary bacterium ferrum element; Pazelliptine; Pegaspargase; Peldesine; The many sodium sulfate of pentosan; Pentostatin; Pentrozole; Perflubron; Perfosfamide; Perillyl alcohol (perillyl alcohol); Phenazinomycin; Phenylacetate; Inhibitors of phosphatases; Streptococcus hemolyticus; Pilocarpine hydrochloride; Pirarubicin; Piritrexim; Placetin A; Placetin B; The plasminogen activator inhibitor; Platinum complexes; Platinum compounds; Platinum-three amine compound; Porfimer sodium; Porfiromycin; Prednisone; Propyl group two acridones; Prostaglandin J2; Proteasome inhibitor; Immunomodulator based on protein A; Inhibitors of protein kinase C; Inhibitors of protein kinase C, microalgae (microalgal); Protein tyrosine phosphatase inhibitor; Purine nucleosides phosphine enzyme inhibitor; Alizarinopurpurin; The pyrazolo acridine; Many oxyalkylenes of myocorilization (pyridoxylated) hemoglobin conjugate; The raf antagonist; Raltitrexed; Ramosetron; Ras farnesyl-protein transhipment enzyme inhibitor; The ras inhibitor; The ras-GAP inhibitor; The demethyl retelliptine; Rhodium Re 186 etidronate; Rhizomycin; Ribozyme; The RII thunder is for amide; Rogletimide; Rohitukine; The Luo Namo peptide; Roquinimex; Rubiginone B1; Ruboxyl; Safingol; Saintopin; SarCNU; Sarcophytol A; Sargramostim; Sdi 1 analogies; Semustine; Old and feeble deutero-inhibitor 1; Positive MODN; Signal transduction inhibitor; Signal transduction modulators; Single chain antigen binding protein; Sizofiran; Sobuzoxane; Boron [10B] card sodium; Phenyl acetic acid is received; Solverol; SM-binding protein; Sonermin; Sparfosic acid; Spicamycin D; Spiromustine; Splenopentin; Sponge element 1; Zamene amine; Stem cell inhibitors; The stem cell division inhibitor; Stipiamide; The substrate degradation enzyme inhibitor; Sulfinosine; Superactivity vasoactive peptide antagonists; Suradista; Suramin; Sphaerophysine; Synthetic glycosaminoglycan; Tallimustine; The tamoxifen methiodide; Tauromustine; Tazarotene; Tecogalan sodium; Ftorafur; Tellurapyrylium; Telomerase inhibitor; Temoporfin; The temozolomide; Teniposide; Tetrachlorodecaoxide; Tetrazolium amine; Thaliblastine; Thiocoraline (thiocoraline); Thrombopoietin; Thrombopoietin mimetics; Thymalfasin; The thymopoietin receptor stimulating agent; Thymotrinan; Thyrotropin; Etioporphyrin (ETIO) ethyl stannum; Tirapazamine; Dichloro titanium alkene; Topsentin; Toremifene; The myeloid-lymphoid stem cell factor; Translational inhibitor; Retinoic acid; Triacetyl uridine; Triciribine; Trimetrexate; Triptorelin; Tropisetron; Turosteride; Tyrosine kinase inhibitor; Tyrphostin (tyrphostins); The UBC inhibitor; Ubenimex; The deutero-growth inhibiting factor of genitourinary system hole; The urokinase receptor antagonist; Vapreotide; Variolin B; Carrier system, the erythrocyte gene therapy; Velaresol; Vermine; Verdins; Verteporfin; Vinorelbine; Vinxaltine; Vitaxin; R 83842; Zanoterone; Zeniplatin; The benzal dimension; With the neocarzinostain NCS ester.

Cancer therapy drug can be chemotherapeutics, and it includes but not limited to following chemical compound: cytotoxic antibiotics, antimetabolite, antimitotic drug, alkylating agent, platinum compounds, arsenic compound, DNA topoisomerase enzyme inhibitor, taxone (taxanes), nucleoside analog, plant alkaloid and toxin; And synthesis of derivatives.Table 1 has been enumerated exemplary chemical compound:

Table 1

Alkylating agent

Nitrogen mustards: cyclophosphamide

Ifosfamide

Trofosfamide

Chlorambucil

Nitroso ureas: carmustine (BCNU)

Luo Mositing (CCNU)

Alkylsulfonate: busulfan

Treosulfan

Triazenes: dacarbazine

Contain platinum compounds: cisplatin

Carboplatin

Ah network's platinum (Aroplatin)

Oxaliplatin

Plant alkaloid

Vinca alkaloids: vincristine

Vinblastine

Vindesine

Vinorelbine

Taxanes: paclitaxel

Docetaxel

The DNA topoisomerase enzyme inhibitor

Epipodophyllins: etoposide

Teniposide

Topotecan

9-aminocamptothecin

Camptothecine

Crisnatol

Mitomycin: ametycin

Anti-folic acid class:

DHFR inhibitor: methotrexate

Trimetrexate

IMP dehydrogenase inhibitor: mycophenolic acid

Thiazole furan quinoline

Ribavirin

EICAR

Ribonucleotide reductase inhibitor: hydroxyurea

Deferoxamine

Pyrimidine analogue

Uracil analogues: 5-fluorouracil

Fluorine urine deoxynucleoside

Doxifluridine

Raltitrexed

Cytosine analog: cytosine arabinoside (ara C)

The cytosine cytosine arabinoside

Fludarabine

Purine analogue: mercaptopurine

2-amino-Ismipur

DNA antimetabolite: 3-HP

2 '-deoxidation-5-fluorouracil

5-HP

α-TGDR

The glycine aphidicolin

ara-C

5-azepine-2 '-the deoxidation cytosine

β-TGDR

Ancitabine

Guanazole

Hypoxanthine sugar dialdehyde

macebecin?II

The pyrazolo imidazoles

Antimitotic agent: allocolchicine

Halichondrins (Halichondrin B)

Colchicine

Colchicine derivative

dolstatin?10

Maytansine

Rhizomycin

Muscoril

The trityl cysteine

Other:

The isoprenylation inhibitor:

Dopaminergic neurotoxin: 1-methyl-4-phenylpyridinium  ion

Cell cycle inhibitor: staurosporine (Staurosporine)

D actinomycin D: actinomycin D

Dactinomycin

Bleomycin: bleomycin A2

Bleomycin B2

Peplomycin

Anthracycline antibiotics: daunorubicin

Doxorubicin (amycin)

Idarubicin

Epirubicin

Pirarubicin

Zorubicin

Mitoxantrone

MDR inhibitor: verapamil

Ca ²⁺Atpase inhibitor: thapsigargin

The present invention has also considered to comprise the compositions of one or more chemotherapeutics (as FLAG, CHOP).FLAG comprises fludarabine, cytosine arabinoside (Ara-C) and G-CSF.CHOP comprises cyclophosphamide, vincristine, doxorubicin and prednisone.Above-mentioned what enumerate all is illustrative, rather than restrictive.

In one embodiment, breast carcinoma can be used the medicine composite for curing of the combination that comprises complex of the present invention and 5-fluorouracil, cisplatin, docetaxel, doxorubicin, Herceptin , gemcitabine, IL-2, paclitaxel and/or VP-16 (etoposide).

In another embodiment, carcinoma of prostate can be used the medicine composite for curing of the combination that comprises complex of the present invention and paclitaxel, docetaxel, mitoxantrone and/or androgen receptor antagonists (as flutamide).

In another embodiment, leukemia can with comprise complex of the present invention and fludarabine, cytosine arabinoside, gemtuzumab (gemtuzumab) (MYLOTARG), the medicine composite for curing of the combination of daunorubicin, methotrexate, vincristine, Ismipur, idarubicin, mitoxantrone, etoposide, asparaginase, prednisone and/or cyclophosphamide.As another example, myeloma can be used the medicine composite for curing of the combination that comprises complex of the present invention and dexamethasone.Preferably, leukemia is chronic myeloid leukemia (CML), and the HSP complex comprises the hsp70-peptide complexes, and the treatment pattern is that methanesulfonic acid is according to imatinib or Gleevec ^TM

In another embodiment, melanoma can be used the medicine composite for curing of the combination that comprises complex of the present invention and dacarbazine.

In another embodiment, colorectal carcinoma can use the medicine composite for curing of the combination that comprises complex of the present invention and irinotecan.

In another embodiment, pulmonary carcinoma can be used the medicine composite for curing of the combination that comprises complex of the present invention and paclitaxel, docetaxel, etoposide and/or cisplatin.

In another embodiment, non-Hodgkin lymphoma can use the medicine composite for curing of the combination that comprises complex of the present invention and cyclophosphamide, CHOP, etoposide, bleomycin, mitoxantrone and/or cisplatin.

In another embodiment, gastric cancer can be used the medicine composite for curing of the combination that comprises complex of the present invention and cisplatin.

In another embodiment, cancer of pancreas can use the medicine composite for curing of the combination that comprises complex of the present invention and gemcitabine.

According to the present invention, can be before cancer therapy drug, use complex of the present invention afterwards or simultaneously, be used for prevention or treatment cancer.According to the type of cancer, medical history and the situation and the selected cancer therapy drug of main body, the use of complex of the present invention and the dosage and the time of chemotherapy are coordinated mutually.

Use complex of the present invention to append in the chemotherapy regimen.In one embodiment, chemotherapeutics be dosage from 100 to 1000mg/m ²The gemcitabine in/cycle.In one embodiment, chemotherapeutics be dosage from 200 to 4000mg/m ²The dacarbazine in/cycle.In preferred embodiments, the dosage of dacarbazine is 700 to 1000mg/m ²/ the cycle.In another embodiment, chemotherapeutics be dosage from 25 to 50mg/m ²The fludarabine in/cycle.In another embodiment, chemotherapeutics be dosage from 200 to 2000mg/m ²The cytosine arabinoside in/cycle (Ara-C).In another embodiment, chemotherapeutics is the docetaxel of dosage from 1.5 to 7.5mg/kg/ cycles.In another embodiment, chemotherapeutics is the paclitaxel of dosage from 5 to 15mg/kg/ cycles.In another embodiment, chemotherapeutics is the cisplatin of dosage from 5 to 20mg/kg/ cycles.In another embodiment, chemotherapeutics is the 5-fluorouracil of dosage from 5 to 20mg/kg/ cycles.In another embodiment, chemotherapeutics is the doxorubicin of dosage from 2 to 8mg/kg/ cycles.In another embodiment, chemotherapeutics is the etoposide of dosage from 40 to 160mg/kg/ cycles.In another embodiment, chemotherapeutics is the cyclophosphamide of dosage from 50 to 200mg/kg/ cycles.In another embodiment, chemotherapeutics is the irinotecan in dosage from 50 to 75,75 to 100,100 to 125 or 125 to 150mg/kg/ cycles.In another embodiment, chemotherapeutics is the vinblastine in dosage from 3.7 to 5.4,5.5 to 7.4,7.5 to 11 or 11 to 18.5mg/kg/ cycles.In another embodiment, chemotherapeutics is that dosage from 0.7 to 1.4 or 1.5 is to 2mg/m ²The vincristine in/cycle.In another embodiment, chemotherapeutics is that dosage from 3.3 to 5,5 to 10,10 to 100 or 100 is to 1000mg/m ²The methotrexate in/cycle.

In a preferred embodiment, the present invention also comprises the chemotherapeutics that the uses low dosage part as combined treatment.For example, using complex of the present invention to carry out initial treatment has increased tumor to the sensitivity of the attack of the chemotherapeutics of doses subsequently, and this dosage is near or below the lower limit of the chemotherapeutics dosage of using when not having complex of the present invention.

In one embodiment, complex of the present invention and low dosage (are arrived 60mg/m as 6 ²/ sky or lower) docetaxel is used the cancer patient.In another embodiment, complex of the present invention and low dosage (are arrived 135mg/m as 10 ²/ sky or lower) paclitaxel is used the cancer patient.In another embodiment, complex of the present invention and low dosage (are arrived 25mg/m as 2.5 ²/ sky or lower) fludarabine is used the cancer patient.In another embodiment, complex of the present invention and low dosage (are arrived 1.5g/m as 0.5 ²/ sky or lower) cytosine arabinoside (Ara-C) is used the cancer patient.In another embodiment, chemotherapeutics is a dosage range 10 to 100mg/m ²The gemcitabine in/cycle.In another embodiment, chemotherapeutics is that dosage from 5 to 10,10 to 20,20 to 40 or 40 is to 75mg/m ²The cisplatin in/cycle is as PLATINOL or PLATINOL-AQ (Bristol Myers).In another embodiment, with 7.5 to 75mg/m ²The cisplatin of the dosage in/cycle is used bladder cancer patients.In another embodiment, with 5 to 50mg/m ²The cisplatin of the dosage in/cycle is used bladder cancer patients.In another embodiment, chemotherapeutics is 2 to 4,4 to 8,8 to 16,16 to 35 or 35 to 75mg/m ²The carboplatin in/cycle is as PARAPLATIN (Bristol Myers).In another embodiment, ovarian cancer patients is used 7.5 to 75mg/m ²The carboplatin in/cycle.In another embodiment, bladder cancer patients is used 5 to 50mg/m ²The carboplatin of the dosage in/cycle.In another embodiment, the carcinoma of testis patient is used 2 to 20mg/m ²The carboplatin in/cycle.In another embodiment, chemotherapeutics is that dosage from 6 to 10,10 to 30 or 30 is to 60mg/m ²The Docetaxel in/cycle is as TAXOTERE (Rhone Poulenc Rorer).In another embodiment, chemotherapeutics is that dosage from 10 to 20,20 to 40,40 to 70 or 70 is to 135mg/m ²The paclitaxel in/cycle is as TAXOL (Bristol Myers Squibb).In another embodiment, chemotherapeutics be dosage from 0.5 to 5mg/m ²The 5-fluorouracil in/cycle.In another embodiment, chemotherapeutics is that dosage from 2 to 4,4 to 8,8 to 15,15 to 30 or 30 is to 60mg/m ²The doxorubicin in/cycle is as ADRIAMYCIN (Pharmacia﹠amp; Upjohn), DOXIL (Alza), RUBEX (Bristol Myers Squibb).

In another embodiment, complex of the present invention and one or more angiogenesis inhibitor medicine administering drug combinations, it includes but not limited to, angiotensin, thalidomide, kringle 5, endostatin, Serpin (serpin) anticoagulin, the 29kDa amino terminal of fibronectin and the proteolytic fragments of 40kDa carboxyl terminal, the 16kDa proteolytic fragments of prolactin antagonist, the 7.8kDa proteolytic fragments of PF4, segmental 13 amino acid whose peptides (people such as Maione corresponding to PF4,1990, Cancer Res., 51:2077-2083), segmental 14 amino acid whose peptides (people such as Tolma corresponding to type i collagen, 1993, J.Cell Biol., 122:497-511), segmental 19 amino acid whose peptides (people such as Tolsma corresponding to thrombosthenin I, 1993, J.Cell Biol., 122:497-511), corresponding to SPARC segmental 20 amino acid whose peptides (people such as Sage, 1995, J.Cell.Biochem., 57:1329-1334), or any fragment, the family member, or its variant, comprise the acceptable salt of its pharmacy.

Also described suppress angiogenesis and corresponding to segmental other peptide of laminin, fibronectin, precollagen and EGF (referring to for example Cao, 1998, Prog Mol SubcellBiol., 20:161-176).Proved that blocking-up has anti-angiogenic activity (people such as Brooks, 1994, Science, 264:569-571 in conjunction with the monoclonal antibody and the ring-type pentapeptide (promptly having peptide motif Arg-Gly-Asp) of proteic some integrin of RGD; People such as Hammes, 1996, Nature Medicine, 2:529-533).In addition, the receptor antagonist body suppresses upar and has suppressed angiogenesis, tumor growth and transfer (people such as Min, 1996, Cancer Res., 56:2428-33; People such as Crowley, 1993, Proc Natl AcadSci., 90:5021-25).The present invention has also considered the applied in any combination of this anti-angiogenic medicaments and complex.

In another embodiment, complex of the present invention and hormone therapy combination is used.Hormone therapy comprises the hormone agonist, hormone antagonist is (as flutamide, bicalutamide, tamoxifen, Lei Luoxifen, leuprorelin acetate (LUPRON), the LH-RH antagonist), the inhibitor of hormone biosynthesis and processing and steroidal are (as dexamethasone, retinoid, deltoids, betamethasone, hydrocortisone, cortisone, prednisone, the 2-boldenone, glucocorticoid, mineralocorticoid, estrogen, Testosterone, progesterone), vitamin A derivative (as all-trans vitamin A acid (ATRA)); Vitamin D 3 analogs; Anti-lutein (as Mifepristone, onapristone) and androgen antagonist (as cyproterone acetate).

In another embodiment, complex of the present invention is used for making up with the gene therapy procedure of treatment of cancer.In one embodiment, will secrete the gene therapy and the complex of the present invention combination of the reconstitution cell of interleukin-2, be used for prevention or treatment cancer, particularly breast carcinoma (referring to people such as for example Deshmukh, 2001, J.Neurosurg., 94; 287-92).In other embodiments, use the polynucleotide chemical compound to carry out gene therapy, the polynucleotide chemical compound is such as but not limited to antisense polynucleotides, ribozyme, rnai molecule, triple helical polynucleotide or the like, and wherein the nucleotide sequence of the DNA of the nucleotide sequence of this chemical compound and generation, development and/or the pathology related gene of tumor or cancer and/or RNA is relevant.For example, they are many to be oncogene, growth factor gene, growth factor receptor gene, cell cycle gene, DNA-repair gene, and is as known in the art.

In another embodiment, complex of the present invention is used with the radiotherapy scheme.For radiotherapy, lonizing radiation can be gamma-rays or X-ray.This method comprises the treatment of cancer that relates to radiotherapy, as the radiotherapy of the treatment of the tissue space implantation of outer beam radiation treatments, radiosiotope (I-125, palladium, iridium), radiosiotope such as strontium-89, chest radiotherapy, intraperitoneal phosphorus-32 radiotherapy and/or full abdominal part and pelvis.For the comprehensive review of radiotherapy, referring to Hellman, the 16th chapter: Principlesof Cancer Management:Radiation Therapy, sixth version, 2001, people such as DeVita write, J.B.Lippencott Company, Philadelphia.In preferred embodiments, radiotherapy is carried out as external beam radiation or teletherapy, and wherein radiation is directly from remote radioactive source.In a plurality of preferred embodiments, radiotherapy is carried out with internal therapentics or brachytherapy, and wherein radioactive source is positioned at body interior, close cancerous cell or tumor thing.The applied in any combination that also comprises complex of the present invention and photodynamic therapy, described photodynamic therapy comprise photosensitizer administration such as hemoporphyrin and derivant thereof, Vertoporfin (BPD-MA), phthalocyanine, photosensitizer Pc4, de-methoxy hypocrellin A; And 2BA-2-DMHA.

In a plurality of embodiments, the combination of complex of the present invention and at least a chemotherapeutics is used short treatment cycle to the cancer patient of cancer to be treated.Use the persistent period of chemotherapeutics treatment can be according to the difference of used certain cancer treatment agent and difference.The present invention also considered to be interrupted use or with every day dosage be divided into some aliquots and use.Suitable treatment time for the certain cancer treatment agent can be grasped by those skilled in the art, and the present invention has considered the lasting evaluation to the optimized treatment plan of every kind of cancer therapeutic agent.The present invention has considered wherein to use at least one cycle of single treatment or continuous treatment, preferably more than one-period.Can grasp the suitable length of a treatment cycle by those skilled in the art, and total number of cycles, the interval during week.

In another embodiment, complex of the present invention is used for the chemical compound combination with the side effect (such as but not limited to influenza-like symptom, fever etc.) that improves cancer symptoms (such as but not limited to pain) and produced by complex of the present invention.Therefore, the chemical compound lot of known minimizing pain, influenza-like symptom and fever can be used for making up with complex of the present invention or being used for mixing with complex of the present invention.This chemical compound comprises analgesic (as acetaminophen), decongestant (as pseudoephedrine), antihistaminic (as chlorphenamine maleate) and antitussive (as dextromethorphan).

5.5. target infectious disease

Infectious disease that can be by method of the present invention treatment or prevention is by including but not limited to that virus, antibacterial, fungus, protozoacide, anthelmintic and parasitic infective agent cause.The invention is not restricted to treat or prevent by in the born of the same parents or the infectious disease that causes of the outer pathogen of born of the same parents.Therapeutic alliance comprises also uses one or more patterns that help to prevent or treat infectious disease except using pharmaceutical composition of the present invention, this pattern includes but not limited to antibiotic, antiviral agents, protozoacide chemical compound, antifungal compound and anthelmintic.Other treatment pattern that can be used for treating or keep off infection comprises aforesaid immunotherapeutic agent, polynucleotide, antibody, cytokine and hormone.

The infective virus of people and non-human vertebrate comprises retrovirus, ribonucleic acid virus and deoxyribonucleic acid virus(DNA virus).The example of the virus of having found in the people includes but not limited to: Retroviridae (as the human immunodeficiency virus, (is also referred to as HTLV-III, LAV or HTLV-III/LAV or HIV-III as HIV-1; With other separator such as HIV-LP; Picornaviridae is (as poliovirus, hepatitis A virus; Enterovirus, human coxsackievirus, rhinovirus, ECHO virus); The Caliciviridae bacterial strain of gastroenteritis (as cause); Togaviridae (as equine encephalitis virus, rubella virus); Swine fever virus belongs to (as dengue virus, encephalitis, yellow fever virus); Coronaviridae (as coronavirus); Rhabdoviridae (as vesicular stomatitis virus, rabies virus); Line Viraceae (as Ebola virus); Paramyxoviridae (as parainfluenza virus, mumps virus, Measles virus, respiratory syncytial virus); Orthomyxovirus section (as influenza virus); Bunyaviridae (as the smooth sick element of the Chinese, parasitics Herba aeginetiae indicae virus, sand fly virus (phleboviruses) and Nairo virus); Husky Viraceae (Arena viridae) (hemorrhagic fever virus); Reoviridae (as reovirus, Orbivirus (orbiviurses) and rotavirus); Double-core ribonucleic acid virus section; Hepatovirus section (hepatitis B virus); Parvoviridae (parvovirus); Papovaviridae (papillomavirus, polyoma virus); Adenoviridae (most of adenovirus); Herpetoviridae (herpes simplex virus (HSV) 1 and 2, varicella zoster virus, cytomegalovirus (CMV), herpesvirus; Poxviridae (smallpox virus, vaccinia virus, poxvirus); And Iridoviridae (as African swine fever virus); ((vehicle of thinking hepatitis B virus defective appurtenance, non-A non-B hepatitis (is propagated in 1 class=body as the pathogen of spongiform encephalopathy, the pathogen of hepatitis D with non-classified virus; 2 classes=non-intestinal transmitted (being hepatitis C); Norwalk and correlated virus and Astrovirus).

Above-mentioned retrovirus comprises simple retrovirus and complicated retrovirus.Simple retrovirus comprises Type B retrovirus, C type retrovirus and D type retrovirus subgroup.The example of Type B retrovirus is mouse mammary adenoma virus (MMTV).C type retrovirus comprises C type A group subgroup (comprising rous sarcoma virus (RSV), avian leukosis virus (ALV) and avian meloblastosis virus (AMV)) and C type B group subgroup (comprising Muridae leukemia virus (MLV), cat leukemia virus (FeLV), murine sarcoma virus (MSV), Gibbon leukemia virus (GALV), SNV (SNV), avian reticuloendotheliosis virus (RV) and ape sarcoma virus (SSV)).D type retrovirus comprises Mei-Pa monkey disease poison (MPMV) and 1 type ape retrovirus (SRV-1).Complicated retrovirus comprises slow virus, T chronic myeloid leukemia virus and foamy virus subgroup.Slow virus comprises HIV-1, and comprises HIV-2, SIV, visna virus, cat family immunodeficiency virus (FIV) and equine infectious anemia virus (EIAV).T chronic myeloid leukemia virus comprises HTLV-1, HTLV-II, ape T chronic myeloid leukemia poison (STLV) and bovine leukemia virus (BLV).Foamy virus comprises Human foamy spumavirus (HFV), ape foamy virus (SFV) and bovine foamy virus (BFV).

Example as the antigenic ribonucleic acid virus of vertebrates includes but not limited to following: the Reoviridae member comprises positive reovirus genus (many serotypes of the retrovirus of mammal and birds), Orbivirus ((blue tongue virus, Eugenangee virus, Kemerovo virus, african horse sickness virus and colorado tick fever virus), rotavirus (Human reoviruslike agent, nebraska calf diarrhea virus, Muridae rotavirus, ape rotavirus, cattle or sheep rotavirus, birds rotavirus); The Picornaviridae member comprises that (poliovirus, CA and B, intestinal cell pathological changes orphan (ECHO) virus, hepatitis A virus, ape enterovirus, Muridae encephalomyelitis (ME) virus, poliovirus muris, bovine enteroviruses, pig enterovirus, cardiovirus (encephalomyocarditis virus (EMC), encephalomyocardis virus), (ERC group virus comprises at least 113 hypotypes to Rhinovirus to enterovirus genus; Other rhinovirus), Hostis (foot and mouth disease (FMDV); The Caliciviridae member comprises pig blister sex vesicle exanthema virus, San Miguel sea lion virus, cat family picorna virus and Norwalk virus; The Togaviridae member comprises alphavirus (eastern equine encephalitis virus, Semliki Forest virus, sindbis alphavirus, the chikungunya fever virus, Ao Niwengniweng virus, ross river virus, Venezuelan equine encephalitis virus, western equine encephalitis virus), mosquito-borne Tobamovirus (mosquito-borne yellow fever virus, dengue virus, Japanese encephalitis virus, Saint Louis' encephalitis virus, Austrian X-disease virus, west Nile virus, KUN, Central European tick-borne virus, the tick-borne virus in the Far East, Kyasanur forest virus, Louping III virus, Bo Wasang virus, msk haemorrhagia fever virus), rubella virus genus (rubella virus), pestivirus (bovine diarrhoea virus, swine fever virus, border disease virus); The Bunyaviridae member comprises that Bunyvirus belongs to (Bunyan Wella and correlated virus, galifornia encephalitis papova), Phlebovirus belongs to (sandfly fever Sicily virus, Rift valley fever virus), Nairovirus (crimean-Congo hemorrhagic fever virus, nairobi sheep disease virus) and Uukuvirus and belongs to (Uukuniemi and correlated virus); The member of orthomyxoviridae family comprises Influenza Virus (influenza virus A type, various human hypotype); Swine influenza virus and birds and equine influenza virus; Influenza Type B (various human hypotype) and influenza C type (may be independent genus); The member of Paramyxoviridae section comprises that paramyxovirus genus (1 type parainfluenza virus, Sendai virus, hemadsorption virus, 2 to 5 type parainfluenza viruses, new city eqpidemic disease virus, mumps virus), Morbillivirus (Measles virus, SSPE cirus, distemper virus, rinderpest virus), pneumonitis virus belong to (respiratory syncytial virus (RSV), bovine respiratory syncytial virus and mouse pneumonia virus); Forest virus, sindbis alphavirus, Chikungunya virus, Ao Niwengniweng virus, ross river virus, Venezuelan equine encephalitis virus, western equine encephalitis virus), mosquito-borne Tobamovirus (mosquito-borne yellow fever virus, dengue virus, Japanese encephalitis virus, Saint Louis' encephalitis virus, Austrian X-disease virus, west Nile virus, KUN, Central European tick-borne virus, the tick-borne virus in the Far East, Kyasanur forest virus, Louping III virus, Bo Wasang virus, msk haemorrhagia fever virus), rubella virus genus (rubella virus), pestivirus (bovine diarrhoea virus, swine fever virus, border disease virus); The Bunyaviridae member comprises that Bunyvirus belongs to (Bunyan Wella and correlated virus, galifornia encephalitis papova), Phlebovirus belongs to (sandfly fever Sicily virus, Li Fute valley fever virus), Nairovirus (crimean-Congo hemorrhagic fever virus, nairobi sheep disease virus) and Wu hole (Uukuvirus) genus (Uukuniemi and correlated virus); The member of orthomyxoviridae family comprises Influenza Virus (influenza virus A type, various human hypotype); Swine influenza virus and birds and equine influenza virus; Influenza Type B (various human hypotype) and influenza C type (may be independent genus); The member of Paramyxoviridae section, comprise paramyxovirus genus (1 type parainfluenza virus, Sendai virus, hemadsorption virus, 2 to 5 type parainfluenza viruses, Avian pneumo-encephalitis virus, mumps virus), Morbillivirus (Measles virus, SSPE cirus, distemper virus, rinderpest virus), pneumonitis virus belongs to (respiratory syncytial virus (RSV), bovine respiratory syncytial virus and mouse pneumonia virus), the Rhabdoviridae member comprises that Vesiculovirus belongs to (VSV), chandipura virus, Flanders-Hart Park virus), Lyssavirus (rabies virus), fish rhabdovirus and two kinds of possible rhabdoviruses (Marburg virus and Ebola virus); The Arenaviridae member comprises the sick virus of lymphocytic choriomeningitis virus (LCM), tacaribe virus complex and lassa fever; The member of Coronoaviridae section comprises infectious bronchitis virus (IBV), Mouse hepatitis virus, people's enteric coronavirus virus and feline infectious peritonitis (cat family coronavirus).

For the example of the antigenic illustrative ribonucleic acid virus of vertebrates includes but not limited to following: Poxviridae comprises orthopoxvirus (variola major, variola minor, monkeypox, cowpox, the Babalus bubalis L. pox, rabbit variola, ectromely), rabbitpox virus belongs to (myxoma, fibroma), Avipoxvirus (fowl pox, other birds poxvirus), Capripoxvirus (sheep pox, goatpox), Suipoxvirus (swine pox), parapoxvirus belongs to (infectiousness papilla dermatitis virus, pseudocowpox, ulcerative stomatitis of cattle virus), Iridoviridae section (African swine fever virus, frog virus 2 and 3, the lymphocystis disease virus of fish); Herpetoviridae, comprise α herpesvirus (herpes simplex 1 and 2 types, varicella-zoster, equine abortion virus, equid herpesvirus 2 and 3, pseudorabies virus, infectious bovine keratoconjunctivitis virus, infectious bovine rhinotrachetis virus, feline rhinotracheitis virus, infectious laryngotracheitis virus) β-herpesvirus (cytomegalovirus of human cytomegalic inclusion disease virus and pig, monkey and Rodents); Gamma herpes viruses (Epstein-Barr virus (EBV), marek's disease virus, Squirrel monkey virus, herpesvirus ateles, herpesvirus Sylvilagus, GPHV, frog herpesvirus); Adenoviridae comprises mastadenovirus (people's A, B, C, D, E and ungrouped subgroup; The adenovirus of simian adenovirus (at least 23 kinds of serotype), infectious canine hepatitis and cattle, pig, sheep, the frog and many other species, Aviadenovirus (aviadenovirus); With the adenovirus that can not raise and train; Papoviridae section, comprise Papillomavirus (the multiple morbific human papillomavirus of human papillomavirus, bovine papilloma virus, Xiao Pu rabbit papilloma virus and other species), Polyomavirus (polyoma virus, simian virus 40 (SV-40), vacuolating virus of rabbit (RKV), K virus, BK virus, JC virus and other primate polyoma virus such as lymphotropic (lymph) human papillomavirus); Parvoviridae comprises that adeno-associated virus (AAV) belongs to, parvovirus belongs to (the full leukopenia syndrome virus of cat, bovine parvovirus, Canine Parvovirus, Aleutian mink disease virus etc.).At last, DNA viruses can comprise virus such as kuru virus and creutzfeldt-Jacob disease virus and the chronic infection neuropathy vehicle that does not meet above-mentioned section.

The many examples that can be used for the antiviral compound of complex of the present invention combination are that this area kind is known, and it includes but not limited to: rifampicin, nucleoside reverse transcriptase inhibitor (as AZT, ddI, ddC, 3TC, d4T), non-nucleoside reverse transcriptase inhibitor (as efavirenz, nevirapine), protease inhibitor (as aprenavir, indinavir, ritonavir and Saquinavir), idoxuridine, cidofovir, acyclovir, ganciclovir, zanamivir, amantadine and palivizumab (Palivizumab).Other example of antiviral agents includes but not limited to acemannan (Acemannan); Acyclovir; Acycloguanosine sodium; Adefovirdipivoxil; Alovudine; Alvircept sudotox; Amantadine hydrochloride; Aranotin; Arildone; The methanesulfonic acid atevirdine; Avridine; Former times many good fortune Wei; Cipamfylline; Cytarabine hydrochloride; Methanesulfonic acid De Laweiding; Desciclovir; Didanosine; Two  are husky sharp; Edoxudine; Enviradene; Enviroxime; Famciclovir; The hydrochloric acid Famotidine; Fiacitabine; Fialuridine; Fosarilate; Foscarnet sodium; Fosfonet Sodium; Ganciclovir; Ganciclovir sodium; Idoxuridine; Deoxidation butanone aldehyde; Lamivudine; Lobucavir; Memotine hydrochloride; Methisazone (Methisazone); Nevirapine; Penciclovir; Pirodavir; Not adept Wei Lin; Rimantadine hydrochloride; Saquinavir mesilate; Somantadine hydrochloride; Sorivudine; Statolon (statolon); Take charge of his furan pyridine; Tilorone hydrochloride; Trifluorothymidine; Valaciclovir hydrochlordide; Vidarabine; Vidarabine phosphate; The vidarabine disodium hydrogen phosphate; Viroxime; Zalcitabine; Zidovudine; Zinviroxime.

Can cause that it includes but not limited to have the antibacterial in born of the same parents' endoparasitism stage such as mycobacteria (as pulmonary tuberculosis mycobacteria (Mycobacteria tuberculosis), cow mycobacteria (M.Bovis), bird mycobacterium (M.Avium), leprosy by the bacterial infection or the disease of method of the present invention treatment or prevention in its life cycle by antibacterial

Mycobacteria (M.Leprae) or African tulase (M.Africanum)), rickettsia, mycoplasma, chlamydia and legionella.Other example of the bacterial infection of considering includes but not limited to by Gram-positive bacillus (as Listeria (Listeria, Bacillus) as anthrax bacillus (Bacillus anthracis), erysipelothrix (Erysipelothrixspecies)), gram negative bacilli is (as Bartonella bacillus (Bartonella), brucella (Brucella), Campylobacter (Campylobacter), enterobacteria (Enterobacter), escherich's bacillus (Escherichia), Francisella (Francisella), haemophilus (Hemophilus), Cray Bai Shi bacillus (Klebsiella), morgan's bacillus (Morganella), Bacillus proteus (Proteus), Providence (Providencia), pseudomonas (Pseudomonas), Salmonella (Salmonella), Serratieae (Serratia), Shiga bacillus (Shigella), vibrio (Vibrio) and yersinia (Yersinia)), spirillum antibacterial (dredging spirillum (Borreliaburgdorferi)) as dredging spirillum (Borrelia), comprise the Bo Shi that causes Lyme disease, anaerobe (as actinomycetes (Actinomyces) and clostridium (Clostridium)), Gram-positive and negative cocci, enterococcus (Enterococcus) class, streptococcus (Streptococcus) class, streptococcus pneumoniae (Pneumococcus) class, staphylococcus (Staphylococcus) class, eisseria (Neisseria gonorrhoeae) class.The example of infectious bacteria includes but not limited to helicobacter pylori (Helicobacterpyloris), Bo Shi dredges spirillum (Borelia burgdorferi), have a liking for lung legionella (Legionella pneumophilia), mycobacterium tuberculosis (Mycobacteriatuberculosis), bird mycobacterium (M.Avium), mycobacterium (M.Intracellulare) in the born of the same parents, bear Sa mycobacteria (M.kansaii), the Ge Dengshi mycobacteria, staphylococcus aureus (Staphylococcus aureus), Diplococcus gonorrhoeae (Neisseriagonorrhoeae), Neisseria meningitidis (Neisseria meningitidis), monocytosis Listera (Listeria monocytogenes), streptococcus pyogenes (Streptococcus pyogenes) (A group B streptococcus), streptococcus agalactiae (Streptococcus agalactiae) (B group B streptococcus), Streptococcus viridans (Streptococcus viridans), streptococcus faecalis (Streptococcus faecalis), bargen's streptococcus (Streptococcus bovis), streptococcus pneumoniae (Streptococcuspneumoniae), hemophilus influenza (Haemophilus influenzae), Bacillusantracis, diphtheria corynebacterium (corynebacterium diphtheriae), erysipelothrix ruhsiopathiae (Erysipelothrix rhusiopathiae), clostridium (Clostridium perfringers), clostridium tetani (Clostridium tetani), clostridium perfringen (Enterobacter aerogenes), Cray Bai Shi pulmonitis strain (Klebsiella pneunaomae), pasteurellosis bacillus (Pasturella multocida), Fusobacterium nucleatum (Fusobacterium nucleatum), Streptobacillus moniliformis (Streptobacillus moniliformis), Treponoma palladium (Treponemapallidium), treponenma pertenue (Treponema pertenue), leptospira (Leptospira), rickettsia (Rickettisa) and split the infection that actinomycetes (Actinomyces israelli) cause with color.

Can be used for including but not limited to the antibacterial or the antibiotic of complex combination of the present invention: aminoglycoside antibiotics is (as apramycin; arbekacin; bambermycin; butirosin; dibekacin; neomycin; neomycin; undecylenate; netilmicin; paromomycin; ribostamycin; sisomicin and spectinomycin); the amphenicol antibiotic is (as azidamfenicol; chloromycetin; florfenicol and thiamphenicol); ansamycin antibiotic (as rifamide and rifampicin); carbacephems (as Loracarbef); carbapenems (as biapenem and imipenum); cephalosporin is (as cefaclor; cefadroxil; cefadole; BL-S 640; cefazedone; cefozopran; cefpimizole; cefpiramide and cefpirome); cephamycin is (as cefbuperazone; cefmetazole (cefmetazole) and cefminox); monobactams is (as aztreonam; carumonam and tigemonam); the oxygen cephalo is (as flomoxef; latamoxef); penicillin is (as Amdinocillin; amdinocillin pivoxil; the amoxicillin; bacampicillin; the benzyl penicillanic acid; sodium benzylpenicillin; epicillin; fenbenicillin; flucloxacillin; penamecillin; penethamate hydriodide; penicillin o-benethamine; penicillin 0; penicillin V; penicillin V benzathine benzylpenicillin; breathe out amine penicillin V; penimepicycline and phencihicillin potassium); Lincoln's amine (as clindamycin and cillimycin); macrolide is (as azithromycin; Deltamycin A4; clarithromycin; Roxithromycin; erythromycin and erythromycin acistrate); amfomycin; bacitracin; capreomycin; colistin; enduracidin; tuberactin; tetracycline is (as apicycline; duomycin; clomocyline and demeclocycline); 2,4-di-amino-pyrimidine (as brodimoprim); nitrofuran (as altabactina and furazolium chloride); quinolinones and analog thereof are (as cinoxacin; ciprofloxacin; clinafloxacin; flumequine and grepagloxacin); sulfanilamide is (as the acetyl group sulfalene; benzylsulfamide; noprylsulfamide; phthalylsulfacetamide; sulfachrysoidine and renoquid); sulfone is (as diathymosulfone; glucosulfone sodium; and solapsone); cycloserine; mupirocin and tuberin.

The other example of antibacterial includes but not limited to acedapsone; Vinegar amine acetosulfone sodium; Alamecin; Alexidine; Amdinocillin; Amdinocillin pivoxil; Amicycline; Amifloxacin; Amifloxacin mesilate; Amikacin; Amikacin sulfate; Aminosallcylic acid; Paramisan sodium; The amoxicillin; Amfomycin; The ampicillin; Ampicillin; Apalcillin sodium; Apramycin; Aspartocin; Astromicin sulfate; Avilamycin; Avoparcin; Azithromycin; The azlocillin; Azlocillin sodium; Bacampicillin hydrochloride; Bacitracin; Withered bacitracin methylene disalicylate; Bacitracin zinc; Bambermycin; Calcium benzamidosalicylate; Berythromycin; Betamicin sulfate; Biapenem; Biniramycin; Biphenamine hydrochloride; Bispyrithione magsulfex; Butikacin; The sulphuric acid butirosin; Capreomycin sulfate Capastat sulfate; Carbadox; Carbapen; Carindacillin sodium; Carfecillin sodium; Carbenicillin potassium; Carumonam sodium; Cefaclor; Cefadroxil; Mandokef; Cefamandole nafate; Cefamandole nafate; Cefaparole; Cefatrizine; Cefazaflur sodium; Cefazolin sodium; Cefazolin sodium; Cefbuperazone; Cefdinir; Cefepime; Cefepime hydrochloride; Cefetecol; Cefixime; CefmnenoximeHydrochloride; Cefmetazole; Cefmetazon (Sankyo); Cefonicid list sodium; Cefonicid sodium; Cefoperazone sodium; Ceforanide; Cefotaxime sodium; Cefotetan; Cefotetan Disodium; Cefotiam hydrochloride; Cefoxitin; Cefoxitin sodium; Cefpimizole; Cefpimizole sodium; Cefpiramide; Cefpiramide sodium; Cefpirome Sulfate; Cefpodoxime Proxetil; Cefprozil; Cefroxadine; Cefsulodine sodium; Ceftazidime; Ceftibuten; Ceftizoxime sodium; Ceftriaxone sodium; Cefuroxime; Cefuroxime oxime ester; Cefuroxime pivoxetil; Cefuroxime Sodium; Cephacetrile sodium; Cefalexin; Cefalexin hydrochloride; Cephaloglycin; Cefaloridine; Cephalothin sodium; Cefapirin sodium; Cefradine; Cetotetrine hydrochloride; Cetophenicol; Chloromycetin; Chloramphenicol palmitate; Chloromycetin pantothenic acid complex; Protophenicol (Proto).; Chlorhexidine phosphanilate; Chloroxylenol; Two auric sulfate mycins; Chlortetracycline hydrochloride; Cinoxacin; Ciprofloxacin; Ciprofloxacin; Cirolemycin; Clarithromycin; AM-1091; Clindamycin; Clindamycin Hydrochloride; Clindamycin palmitate hydrochloride; Clindamycin phosphate; Clofazimine; Dry-Clox (Fort Dodge); Cloxacillin sodium; Cloxiquine (Cloxyquin); Colistimethate sodium; Colistin sulfate; Coumamycin; Coumamycin sodium; Cyclacillin; 4-amino-3-isoxazolidone; Dalfopristin (Dalfopristin); Dapsone; Daptomycin; Demecycline; The hydrochloric acid demecycline; Demecycline; Denofungin (Denofungin); Diaveridine; Dicloxacillin; Dicloxacillin sodium; Dihydrostreptomycin sulfate; Dipyrithione; Dirithromycin; Doxycycline; Doxycycline calcium; Doxycycline Fosfatex; Doxycycline hydrochloride; Droxacin sodium; Enoxacin; Epicillin; The hydrochloric acid epitetracycline.; Erythromycin; Erythromycin acistrate; Erythromycin estolate; Erythromycin ethylsuccinate; Erythromycin gluceptate; Erythromycin Lactobionate; Erythromycin propionate lauryl sulfate; Erythromycin octadecanoate; Ebutol; Ethionamide; Fleroxacin; The flucloxacillin; Fludalanine; Flumequine; Fosfomycin; Fosfomycin trometamol; Fumoxicillin; Furazolium chloride; Furazolium tartrate; Sodium fusidate; Ground, husband west; Gentamycin sulfate; Gloximonam; Gramicidin (Gramicidin); Haloprogin; The hetacillin; Hetacin-K (Fort Dodge); Hexedine; Ibafloxacin; Imipenum (Imipenem); Isoconazole; Isepamicin; Isoniazid; Josamycin; Kanamycin sulfate; Kitasamycin; Levofuraltadone; Levopropylcillin potassium; Lexithromycin; Lincomycin; Lincomycin hydrochloride; Lomefloxacin; Lomefloxacin hydrochloride; Lomefloxacin mesilate; Loracarbef; Mafenide; Meclocycline; Traumatociclina; Megalomicin potassium phosphate; Mequidox; Meropenem; Methacycline; Methacycline hydrochloride; Hexamethylenamine; Methenamine hippu; Meng Deli acid hexamethylenamine; Methicillin sodium; Metioprim; The hydrochloric acid metronidazole; The phosphoric acid metronidazole; The mezlocillin; Mezlocillin sodium; U.S. promise tetracycline; The U.S. promise tetracycline of hydrochloric acid; Mirincamycin hydrochloride; Monensin; Rumensin; Sodium nafcillin; Nalidixate sodium; Nalidixan; Natamycin; Nebramycin; Neomycin palmitate; Polygynax; The 9-undecylenic acid neomycin; Netilmicin sulfate; Neutramycin; Nifuradene; Nifuraldezone; Nifuratel; Nifuratrone; Nifurdazil; Nifurimide; Nifurpirinol; Nifurquinazol; Nifurthiazole; Nitrocycline; Nitrofurantoin; Nitromide; Norfloxacin; Novobiocin monosodium; Ofloxacin; Ormetoprim; Oxacillin sodium; Oximonam; Oximonam sodium; Oxolinic acid; Oxytetracycline; Calcium Oxytetracycline.; Tetramycin hydrochloride; Paldimycin; Parachlorophenol; Paulomycin; Pefloxacin; Pefloxacin mesilate; Penamecillin; Penicillin G benzathine benzylpenicillin; Penicillin G potassium; Penicillin G procaine; Penicillin G sodium; Penicillin V; Penicillin V benzathine benzylpenicillin; Breathe out amine penicillin V; Penicillin V potassium; Pentizidone sodium; Phenyl aminosalicylate; Avocin; Pirbenicillin sodium; Piridicillin sodium; Pirlimycin hydrochloride; Pivampicillin hydrochloride; Pivampicillin pamoate; The acid of pivampicillin propyl benzene; Aerosporin; Porfiromycin; Propikacin; Pyrazinamide; Pyrithione zinc; The acetic acid quindecamine; The general fourth of quinoline slave; Racefenicol (Racephenicol); Ramoplanin; Ranimycin; Relomycin; Repromicin; Rifabutin; Rifametane; Rifamexil; Rifamide; Rifampicin; Rifapentine; Rifaximin; Rolitetracycline; Rolitetracycline nitrate; Rosamicin; The butanoic acid rosamicin; The propanoic acid rosamicin; Rosamicin phosphate ester sodium; The stearic acid rosamicin; Rosoxacin; Roxarsone; Roxithromycin; Sancycline; Sanfetrinem sodium; Sarmoxicillin; Sarpicillin; Scopafungin (Scopafingin); Sisomicin; Sisomicin sulfate; Sparfloxacin; The hydrochloric acid grand sight is mould; Spiramycin; Stallimycin Hydrochloride; Steffimycin; Streptomycin sulfate; Streptoniazide (Streptonicozid); Sulfabenz; Sulfabenzamide; Sulfacetamide; Sulphacetamide; Sulfacitine; Sulfadiazine; Sulfadiazine sodium; Sulfadoxine; Sulfalene; Sulfamethyldiazine; Sulfameter; Sulfadimidine; Sulfamethizole; The sulfamethoxazolum azoles; Sulfamonomethoxine; Sulfanilamide  azoles; Sulfanilate zinc; Sulfanitran; Sulfasalazine; Sulfasomizole; Sulfathiazole; Sulfapyrazole; The different  azoles of sulfanilamide; The different  azoles of sulfacetamide; The different  azoles of sulfanilamide diethanolamine salt (Diolamine); Sulfomyxin; Sulopenem; Sultamicillin; Suncillin sodium; Talampicillin hydrochloride; Teicoplanin; The hydrochloric acid temafloxacin; Temocillin; Tetracycline; Quadracycline; Tetracycline phosphate ester complex; Tetroxoprim; Thiamphenicol; Thiphencillin potassium; Ticarcillin cresyl sodium; Ticarcillin disodium; Ticarcillin sodium; Ticlatone; Tiodonium chloride (TiodoniumChloride); Tobramycin; Tobramycin sulfate; Tosufloxacin; Trimethoprim; Trimethoprim sulfate; Neotrizine (Trisulfapyrimidines); Triacetyloleandomycin; Trospectomycin sulfate; Tyrothricin; Vancomycin; Lyphocin (Fujisawa); Virginiamycin; Zorbamycin.

Can include but not limited to aspergillosis, cryptococcosis, sporotrichosis, coccidioidomycosis, Brazilian blastomycosis, histoplasmosis, blastomycosis, zygomycosis and candidiasis by the mycosis of method treatment of the present invention or prevention.

Can be used for dividing antifungal compound to include but not limited to complex combination of the present invention: polyenoid is (as amphotericin B, candicidin, mepartricin, natamycin and nystatin), allylamine (as Boot naphthalene sweet smell and naftifine), imidazoles is (as bifonazole, butoconazole, clodantoin, the fluorine triaconazole, isoconazole, ketoconazole and Lan Nuokang azoles), thiocarbamate is (as tolciclate, tolindate and tolnaftate), triazole is (as fluconazol, itraconazole, Saperconazole and terconazole (triaconazole)), bromosalicylchloranilide, the buclosamide calcium propionate, siccolam, ring pyrrole department, azaserine, griseofulvin, oligomycin, the neomycin undecylenate, pyrrolnitrin, siccayne, tubercidin and viridin.

The other example of antifungal compound includes but not limited to acrisorcin; Ambruticin; Amphotericin B; Azaconazole; Azaserine; Basifungin; Bifonazole; Biphenamine hydrochloride; Bispyrithione magsulfex (Bispyrithione Magsulfex); Nitric acid butoconazole; Calcium undecylenate; Candicidin; Carbol fuchsin; Clodantoin; Ring pyrrole department; Ciclopirox olamine; Cilofungin; Cisconazole; Clotrimazole; Cuprimyxin; Denofungin (Denofungin); Two pyrrole sulfur ; Doconazole; Econazole; Econazole nitrate; Enilconazole; Ethonam nitrate; Fenticonazole nitrate; Filipin (Filipin); Fluconazol; Flucytosine; Fungimycin; Griseofulvin; Hamycin; Isoconazole; Itraconazole; Kalafungin; Ketoconazole; Lomofingin; .alpha.-Dehydrobiotin; Mepartricin; Miconazole; Miconazole nitrate; Monensin; Rumensin; Naftifine hydrochloride; The neomycin undecylenate; Nifuratel; Nifurmerone; The hydrochloric acid nitralamine; Nystatin; Sad; Orconazole nitrate; Oxiconazole Nitrate; Oxifungin hydrochloride; Parconazole hydrochloride; Partricin; Potassium iodide; Proclonol; Pyrithione zinc; Pyrrolnitrin; Rutamycin; Sanguinarium Chloride; Saperconazole; Scopafungin; Selenium sulfide; Sinefungin; Sulconazole nitrate; Terbinafine; Terconazole (triaconazole); Thiram; Ticlatone; Tioconazole; Tolciclate; Tolindate; Tolnaftate; Glyceryl triacetate; Triafuigin; 9-undecylenic acid; Viridoflilvin; Zinc Undecenoate; And zinoconazole hydrochloride.

Can include but not limited to amoebiasis, malaria, leishmaniasis, coccidiosis, giardiasis, latent sorosphere parasitosis, toxoplasmosis and african trypanosomiasis by the parasitic disease of separating method treatment of the present invention or prevention.Also comprise the infection of multiple anthelmintic, such as but not limited to ascariasis, ancylostomiasis, trichuriasis, strongyloidiasis, toxoccariasis, trichonematosis, onchocerciasis, filaricide and dirofilariasis.Also comprise the infection of multiple trematodiasis, such as but not limited to schistosomicide, paragonimiasis westermani and clonorchiasis.Whether the parasite that causes these diseases can be born of the same parents' endoparasitism or the ectoparasite classification of born of the same parents according to them.As used in this article, " born of the same parents entozoa " is that its whole life is intracellular parasite.The entozoal example of plancenta hominis comprises leishmania (Leishnzania spp.), plasmodium (Plasmodium spp.), schizotrypanum cruzi, Mus toxoplasma, Babesia spp. and trichinella." born of the same parents epizoa " is that its whole life is extracellular parasite as used in this article.Born of the same parents epizoa that can infected person comprises entamoeba historlytica, giardia intestinalis, Bi Shi intestinal born of the same parents worms (Enterocytozoonbieneusi), Nai Geli protozoon (Naegleria) and Acanthamoeba (Acantlaamoeba) and most of anthelmintic.Another kind of parasite is defined as and is mainly outside the born of the same parents, but exists in cell in pressure at the critical stage of its life cycle.This parasite is referred to herein as " compulsory born of the same parents entozoa ".These parasites can be spent its most of life or fraction life in born of the same parents' external environment, but have at least one section to force the phase in the born of the same parents in its life cycle.This class parasite of the latter comprises trypanosoma rhodesiense and castellanella gambiense (Trypanosoma gambiense), isospora (Isospora spp.), latent spore coccidiosis (Cryptosporidium spp.), the coccidiosis of liking to be beautiful (Eimeria spp.), new spore worm (Neospora spp.), sarcocystis (Sarcocystis spp.) and schistosomicide (Schistosoma spp.).

The many examples that can be used for the protozoacide chemical compound of complex combined therapy parasitic disease of the present invention are as known in the art, and it includes but not limited to: quinine, chloroquine, mefloquine, proguanil, pyrimethamine, metronidazole, diloxanide, tinidazole, amphotericin, sodium stibogluconate, bactrim and pentamidine.The many examples that can be used for the parasiticide medicine of complex combined therapy parasitic disease of the present invention are as known in the art, and it includes but not limited to: mebendazole, levamisole, niclosamide, praziquantel, albendazole, ivermectin, diethylcarbamazine and thiabendazole.Other example of parasiticide chemical compound includes but not limited to acedapsone; The hydrochloric acid amodiaquine; Amquinate; Arteflene; Chloroquine; Chloroquine hydrochloride; Arechin (Polfa); Camolar; Enpiroline phosphate; The non-alkane of hydrochloric acid chlorine fluorine; Hydroxychloroquine sulfate; Mefloquine Hydrochloride; Menoctone; Mirincamycin hydrochloride; Primaquine phosphate; Pyrimethamine; Quinine sulfate; And tebuquine.

In some embodiments, can use Antybody therapy and/or keep off infection.At one not too in the preferred embodiment, complex of the present invention can be used for non-α 2M be the vaccine combination combination.The example of this vaccine for man is at The Jordan Report 2000, and AcceleratedDevelopment of Vaccines describes among the National Institute of Health to some extent, and it incorporates this paper into as a reference in full.The many vaccines that are used for the treatment of non-human vertebrate are at Bennett, K., and Compendium of Veterinary Products, the third edition, North American Compendiums, Inc. has in 1995 disclosedly, and it incorporates this paper into as a reference in full.

5.6. therapeutic scheme

Be used for the treatment of or any therapeutic alliance of prophylaxis of cancer and infectious disease for above-mentioned, pharmaceutical composition of the present invention can be before united mode be used, use simultaneously or afterwards.Institute's united mode can be above-mentioned be used for the treatment of or the pattern of prophylaxis of cancer or infectious disease in each.

In one embodiment, with pharmaceutical composition of the present invention in reasonably time and another kind of pattern simultaneously to administered.This method regulation is applied in to be less than one minute to about five minutes each other or to be to the maximum in about 60 minutes time range for two kinds to be used, for example, and in once making a house call together.

In another embodiment, use pharmaceutical composition of the present invention and united mode just simultaneously.In another embodiment, pharmaceutical composition of the present invention is used successively with united mode and interval that interests when providing than their independent using increase is used in that pharmaceutical composition of the present invention and pattern can be worked simultaneously.In another embodiment, using of pharmaceutical composition of the present invention and united mode is fully approaching in time, so that required treatment or prevention result to be provided.Can use at the same time or separately with any suitable form with by any suitable approach separately.In one embodiment, pharmaceutical composition of the present invention is used by different route of administration with pattern.In embodiment optionally, the two is used by identical route of administration separately.Pharmaceutical composition of the present invention can be used at identical or different position, as arm and lower limb.When using simultaneously, pharmaceutical composition of the present invention can be with mixture with united mode or is not used with the form of mixture or use at identical position by identical route of administration.

In a plurality of embodiments, complex of the present invention and united mode are used to be less than 1 hour interval, about 1 hour interval, 1 hour to 2 hours interval, 2 hours to 3 hours interval, 3 hours to 4 hours interval, 4 hours to 5 hours interval, 5 hours to 6 hours interval, 6 hours to 7 hours interval, 7 hours to 8 hours interval, 8 hours to 9 hours interval, 9 hours to 10 hours interval, 10 hours to 11 hours interval, 11 hours to 12 hours interval, the interval that is no more than 24 hours interval or is no more than 48 hours.In other embodiments, pharmaceutical composition of the present invention and vaccine combination are used with 2 to 4 days interval, 4 to 6 days interval, the interval in 1 week, the interval in 1 to 2 week, the interval in 2 to 4 weeks, one month interval, 1 to 2 months interval or the interval of period of greater than two months.In preferred embodiments, complex of the present invention and united mode therein the both still have in the active time range and use.Those skilled in the art can determine this time range by detecting each half-life of using component.

In one embodiment, pharmaceutical composition of the present invention and united mode are used in making a house call with once seeking medical advice.In a concrete preferred embodiment, pharmaceutical composition of the present invention was used before described pattern.In specific embodiments optionally, pharmaceutical composition of the present invention is used after described pattern.

In certain embodiments, pharmaceutical composition of the present invention and pattern cycle ground is to administered.Cycle therapy comprises to be used pharmaceutical composition a period of time of the present invention, mode of administration a period of time then, and repeat this sequential and use.Cycle therapy can reduce one or more treatments are formed Drug resistance, avoid or reduce the side effect of wherein a kind of treatment and/or improve therapeutic effect.In this embodiment, the present invention has considered pharmaceutical composition of the present invention after 4 to 6 days, after preferred 2 to 4 days, more preferably alternately use described pattern after 1 to 2 day, and wherein this circulation can repeat arbitrarily repeatedly according to needs.In certain embodiments, pharmaceutical composition of the present invention and pattern be less than 3 weeks, per two weeks once, once or once in a week alternately used in per 10 days.In certain embodiments, the patient used compositions of the present invention in per approximately 1,2,3,4,5,6,7,8,9 or 10 day.

In a specific embodiment, one hour arrives twenty four hours to administered pharmaceutical composition of the present invention after mode of administration.If used the pattern drug-supplying system of slow release or sustained releasing type, then time range can prolong several days at least again.In certain embodiments, as mentioned above, in the whole process in patient's remaining years regularly to administered pharmaceutical composition of the present invention.

5.7.CD91 activity analysis

After its purification, can further characterize α 2M-antigenic molecule complexes, to measure its influence active to CD91 and the CD91 signal transduction pathway.For example, can be by the effect sign α 2M-antigenic molecule complexes of test α 2M-antigenic molecule complexes to the CD91 cytoactive.This analysis comprises antigen-specific, activated analysis of downstream signal transduction analysis, antigen presentation analysis, cytotoxic T cell or the like.This analysis can be used for test and/or measures immunne response.

In a plurality of embodiments, can test the test of α 2M complex to the active effect of congenital CD91 signal transduction.For example, the activatory downstream signal transduction of the CD91 that can analyze includes but not limited to: the moving and chemotaxis (people such as Forrester of the increase of macrophage, 1983, Immunology, 50:251-259), the rising of the synthetic negative adjusting of protease and intracellular Ca2+, inositol monophosphate and cyclic adenosine monophosphate (people such as Misra, 1993, Biochem.J., 290:885-891).Other innate immune responses that can test is the release of cytokine (being IL-12, IL1 β, GMCSF and TNF α).

For example, in one embodiment, can use the chemotaxis test further to characterize from the isolating α 2M of mammalian blood serum complex material standed for.But knownly interact the α 2M inducing cell of modification to its ligand directed moves by protease.Having many technology to can be used for testing external chemotaxis moves (referring to people such as for example Leonard, 1995, " Measurement α and β Chemokines " is in Current Protocols inImmunology, 6.12.1-6.12.28Coligan compile John Wiley﹠amp Deng the people; Sons, Inc., 1995).For example, in one embodiment, the ability of the ability that the cell that can use chemotactic factor gradient test candidate compound to regulate abduction delivering α 2MR receptor in porous Boyden chemotactic chamber moves.In the object lesson of this method, a series of dilutions of the α 2M complex of determining that will be in Preliminary screening place the bottom hole of Boyden chemotaxis case.Also in dilution series, add the constant part.In contrast, at least one waits separatory only to comprise part (as α 2M).By more only comprise part (as α 2M) wait separatory move on the membrane filter lower surface cell quantity with comprise α 2M polypeptide and part (as α 2M) wait cell quantity measurement α 2M complex in the separatory to the contribution of the chemotactic activity of α 2MR.Cause that tested cell number causes that with respect to the solution that use only comprises part (as α 2M) tested cell number reduces, and has then determined the inductive antagonist of part (as α 2M) of the chemotactic activity of express alpha 2M cell if in part (as α 2M) solution, add α 2M complex.

Intracellular ionized calcium ([Ca ²⁺] _i) concentration rises also is the activatory sign of α 2MR (people such as Misra, 1993, the same).Therefore, in another embodiment, the calcium current analysis can be used as the postsearch screening of further sign α 2M complex effect.In the presence of part, be with or without α 2M complex in the presence of measure to express the intracellular calcium concentration of the cell of CD91.For example, can detect and measure calcium current by the fluorescent dye of catching in the flow cytometry labeled cell moves.Fluorescent dye as Indo-1, shows emission spectra and changes in conjunction with calcium the time, the fluorescence that is produced by the calcium combination dye can be used for estimating intracellular calcium concentration to the ratio of the fluorescence that combination dye not produces.In specific embodiment, in comprising the medium of Indo-1 under 37 ℃ in cuvette cultured cell and it is excited, (Photon TechnologyCorporation International) measures fluorescence to use exometer.Be with or without α 2M complex in the presence of add part at particular point in time, in cuvette, add EGTA to discharge and calcium that chelating is whole and measure and reply.The combination of part causes in the cell of express alpha 2MR Ca in the cell ²⁺Concentration increases.Agonist causes Ca in the cell ²⁺Concentration increases relatively, and antagonist causes Ca in the cell ²⁺Concentration reduces relatively.

In other embodiments, in order to detect or measure the activity of α 2M-antigenic molecule complexes, can carry out the antigen presentation test and how have an effect in vivo with prediction α 2M-antigenic molecule complexes.

Thisly present test again for as known in the art, its formerly described (Suto and Srivastava, 1995, Science, 269:1585-1588).For example, in one embodiment, in medium, antigen-presenting cell such as macrophage system (as RAW264.7) are mixed with T cells with antigenic specificity, use about 10,000 cells of each type of about 1: 1 ratio.Add α 2M and the antigenic complex of peptide and cultivated about 20 hours to cell.In one embodiment, IFN-γ release test can be used for measuring or detecting t cell response.After the washing with cell fixation, saturatingization and and can with the antibody of the dye marker of people IFN-gamma reaction (PE-anti--IFN-γ) reaction.Use standard technique by the flow cytometry analytic sample.Perhaps, can use filtration immune detection, ELISA (enzyme linked immunological absorption detects) or enzyme linked immunological trace to detect the specific cell factor of (ELISPOT) detection by the activated T cells generation.In one embodiment, for example, with the cytokine specificity one-level antibody of purification, promptly anti--IFN-γ wraps by the microtitration plate of nitrocellulose backing, and with the background of plate sealing to avoid producing owing to other proteinic non-specific binding.The dilution APC cell sample of antigenic stimulus on the hole of microtitration plate.The adding labelling as biotin labeled secondary anti-cytokine antibodies.Thereby can detect by methods such as for example conjugated streptavidins of enzyme, the cytokine secretion cell can show as " speckle " and detect antibody cytokine complex under vision, microscope or detection of electrons method.In another embodiment, can use " tetramer dyeing " test (people such as Altman, 1996, Science, 274:94-96) identification antigen specific T-cell.For example, will comprise the antigenic MHC molecule of specific peptide poly, and make the soluble peptide tetramer and carry out labelling by for example being compound in streptavidin.Then MHC-peptide antigenic compound is mixed with post-stimulatory T cell colony.Use biotin dyeing T-cell then, the T-cell recognition also is incorporated into the MHC-antigenic compound.

5.8. inducing tumor cell necrosis

In certain embodiments of the invention, before separating α (2) macroglobulin-antigenic molecule complexes from animal mammal the induced tumor necrosis may be favourable.The applicant is verified can to increase the effectiveness of complex in treatment and prophylaxis of cancer in induced tumor necrosis before the isolated complex.Referring to the 6th part embodiment result.The applicant believes, itself is not limited to any theory, downright bad tumor cell release tumor specific antigen.The antigen molecule that discharges enters body fluid such as blood then and forms complex with α (2) macroglobulin.

Many methods can be used for the induced tumor necrosis.The medicine of using the inducing tumor cell necrosis is common in the treatment of cancer.The example of neoplasm necrosis medicine includes but not limited to bleach, cisplatin/epinephrine injectable gelling (people such as Vogl, 2002, British Journal of Cancer, 86 (4): 524-529), ONO-4007, lipid A synthetic analogues (people such as Satoh, 2002, Cancer Immunol.Immunother, 50 (12): 653-662), tumor necrosis factor (TNF), solubility ethanol (people such as Burgener, 1987, Invest Radiol., 22 (6): 472-478), anti-epidermal growth factor receptor (α EGFr) antibody (people such as Wersall, 1997, Cancer Immunol.Immunother., 44 (3): 157-164), OK-432, the lyophilizing streptococcus of handling with penicillin (people such as Ishiko, 1997, Int.J.Immunopharmacol., 19 (7): 405-412), adenovirus (ONYX-015) (people such as Ganly, 2000, Clin.Cancer Res., 6 (3): 798-806), IL4-PE chimeric protein (people such as Rand, 2000, Clin.Cancer Res., 6 (6): 2157-2165), nimustine (ACNU) (people such as Wakabayashi, 2001,18 (1): 23-28).

Also can use other method to realize neoplasm necrosis, such as but not limited to, tumor photosensitization (the people such as Blank who comprises the photodynamic therapy that uses hypercin, 2001, Oncol.Res., 12 (9-10409-418)), United States Patent (USP) 6,131, tissue selectivity microwave irradiation described in 577, United States Patent (USP) 5,928, described in 159 tumor tissues is applied heating with necrosis induced, United States Patent (USP) 5,897, the LASER HEATING of the lower face human body described in 549, United States Patent (USP) 5,895,356 and 5,843, the per urethra focus supersonic therapy of describing in 144 that causes the tumor of prostate necrosis, United States Patent (USP) 5, silver nitrate and dextran are stuck with paste described in 891457 is used for uterus carcinoma to realize the necrosis of body cavity nexine, United States Patent (USP) 5,776, the pulsed magnetic width of cloth described in 175 is penetrated, United States Patent (USP) 5,527, the Non-ionizing radiation described in 352, United States Patent (USP) 5,492, conduit described in 529 with firing equipment, United States Patent (USP) 5,487, endoscope's guiding of the laser energy described in 740, United States Patent (USP) 5,318,564,4,186,729 and 4,237, the device of the usefulness heated by electrodes selectivity tissue described in 898, United States Patent (USP) 5,186,181,4,154,246,4,119,102,3,991,770 and 4,230, the radiofrequency theromtherapy described in 129, United States Patent (USP) 5,159, the heatable inflatable device that is used for the treatment of inner tissue's nexine described in 925, United States Patent (USP) 5,149, the immunopotentiating composition described in 527, United States Patent (USP) 4,983,159 and 4,392,040 and 4,545, near tumor tissues, inject ferromagnetic particle described in 368, United States Patent (USP) 4,763, tissue is applied with conduit described in 671 heated and radiation, with United States Patent (USP) 3, use liquefied gas cooling medium treated tissue described in 674,031.

In certain embodiments, carried out between by two months in necrosis induced can be before separating α (2) macroglobulin complex one hour.In specific embodiment, induce 1 hour, 12 hours, 1 day, 3 days of can be before separating α (2) macroglobulin complex, 1 week, 3 weeks, carry out 5 weeks, 1 month or 2 months.Understandable as skilled doctor, as required, can the superinduce necrosis.

In certain embodiments, before separating α (2) macroglobulin-antigenic molecule complexes, the patient is used chemotherapeutics.In this embodiment, can use those chemotherapeutics described in for example above 5.4 parts.

The list of references of foregoing description induced tumor necrosis is incorporated this paper into as a reference in full.

5.9. dosage regimen and preparation

The covalently or non-covalently complex of α 2M polypeptide of the present invention and antigen molecule can be disposed for the mammal administration, be used for pharmaceutical preparation with the treatment of immunization therapy effective dose or prophylaxis of cancer or infectious disease.Drug dissolution and absorption site factor for when selecting the route of administration of therapeutic agent, considering.

Can in cell culture or laboratory animal, for example detect LD by standard pharmaceutical procedures ₅₀(median lethal dose(LD 50) of colony) and ED ₅₀(the half treatment effective dose of colony) detects this toxicity and the treatment effectiveness that comprises the compositions of α 2M complex.Dosage between toxicity and treatment are renderd a service is than being therapeutic index, and it can be expressed as LD ₅₀/ ED ₅₀Preferably show the alpha2 Macroglobulin complex of big therapeutic index.Though also can use the chemical compound that shows toxic side effects, should note designing drug-supplying system, make this compositions that comprises α 2M complex be oriented to the treated tissue position, thereby the potential damage of non-treatment cell is minimized, reduce side effect thus.

Main body of preferably receiving treatment or patient are mammal, and it includes but not limited to domestic animal, as cat and Canis familiaris L.; Wild beast comprises fox and racoon; Livestock and poultry comprises horse, cattle, sheep, turkey and chicken, and any rodent.Most preferably, main body is behaved.

The data that derive from cell culture test and zooscopy can be used for calculating the multiple dosage of human.The dosage that comprises this compositions of α 2M complex preferably is in and comprises seldom or do not have a toxic ED ₅₀The circulation composition scope in.According to used dosage form and route of administration, dosage can change in this scope.For any compositions of using in the inventive method that comprises α 2M complex, can be at first from cell culture test evaluation treatment effective dose.Can in animal model, calculate a dosage to realize comprising the IC that determines as in the cell culture ₅₀The circulating plasma concentration range of (promptly realizing the concentration of the maximum compositions that comprises α 2M complex that suppresses of half of symptom).This information can be used for determining more accurately human dosage.Blood plasma level can be measured by for example high performance liquid chromatography (HPLC).

The selection of preferred effective dose is determined according to the consideration of the multiple factor known to those skilled in the art by those skilled in the art.This factor comprises the concrete form of the compositions that contains α 2M complex and pharmacokinetic parameter thereof such as bioavailability, metabolism, half-life etc., and it is determined in the used conventional development approach process of the authorities' approval that typically is used for obtaining pharmaceutical compound.The interests that be included in other factors during dosage the is considered situation that will treat in the normal individual and disease maybe will realize; Patient's body weight; Route of administration; Use short-term or secular; Concomitant drugs; With the other factors of known influence institute to medicine effectiveness.Therefore, accurate dose should determine according to doctor's judgement and every patient's situation, as according to the situation of individual patients and immune state, according to the clinical technology of standard.

α of the present invention (2) macroglobulin complex can randomly be used with one or more auxiliary agents, replys with enhance immunity.For example, according to the host type difference, spendable auxiliary agent includes but not limited to: inorganic salt or mineral coagulant such as aluminium hydroxide, aluminum phosphate and calcium phosphate; Surfactant such as LYSOLECITHIN SUNLECITHIN A, polyether polyol (pluronic polyol), polyanion, peptide, oil emulsion, key hole keyhole limpet hemocyanin and dinitrophenol; Molecules of immunization stimulus is as cytokine, Saponin (as QS-21), muramyldipeptide and tripeptide derivative, CpG dinucleotide, CpG oligonucleotide, single phosphatidyl lipid A and polyphosphazene; Granule and micropartical auxiliary agent are as emulsion, liposome, virion, volute casing (cochleates); Or the compound mucosa auxiliary agent of immunostimulation, Freund adjuvant (fully with people's used additives such as BCG (bacillus calmette-guerin vaccine) and coryne bacterium parvum (corynebacterium parvum) incomplete and that may use).In addition, the document of following patent and publication discloses the immunostimulating oligonucleotide that comprises the CpG oligonucleotide that can be used in the present composition: United States Patent (USP) 6,207,646; 6,339,068; 6,239 116; 6,429,199; With the PCT patent disclosure, people's such as Krieg WO 0122972, WO 00/06588; The WO 01/12804 of Agrawal; WO 01/83503, WO 01/55370; People's such as Fearon WO 02/052002; People's such as Tuck WO 01/35991; The WO01/12223 of Van Nest; The WO 99/62923 of Schwartz; WO 98/55495; People's such as Davis United States Patent (USP) 6,406,705; With people's such as Kandimalla PCT patent disclosure WO02/26757, above-mentionedly incorporate this paper into as a reference in full all.Other auxiliary agent that is fit to that can be used among the present invention can be at A Compendium of Vaccine Adjuvants andExcipients (second edition), Vogel, and F., Powell, M. and Alving are among the C.; At Vaccine Design-The Subunit and Adjuvant Approach, Powell, M., Newman, M., Burdman, J., Editors, Plenum Press, New York, 1995, the 141-227 pages or leaves and 2nd Meeting on Novel Adjuvants CurrentlyIn/Close to human Clinical Testing, World HealthOrganization-Organization Mondiale de 1a Sante FoundationMerieux, Annecy, France, 5-7 June 2000, Kenney, R., Rabinovich, N.R., Pichyangkul, S., Price, V., and Engers, H., Vaccine, 20 (2002) 2155-63.It incorporates this paper into as a reference all.

Can use any required route of administration to use α of the present invention (2) macroglobulin complex, it includes but not limited to for example subcutaneous injection, intravenous injection or intramuscular injection, though preferred Intradermal or mucosal administration.The advantage of Intradermal or mucosal administration comprises that respectively the dosage of use is low and absorbs rapidly.That the mucosal approach includes but not limited to is oral, rectum and nasal administration.The preparation of mucosal administration is fit in several formulations as described below.Can in therapeutic process, change route of administration.

Above-mentioned dosage is preferably weekly, uses about 4-6 week, and preferred dosage regimen and position are with use at every turn and change.In a preferred embodiment, adopt subcutaneous administration, the position of at every turn using sequentially changes.Therefore, can be such as but not limited to injection for the first time at the left arm subcutaneous administration, for the second time at right arm, for the third time left abdominal part, the 4th time at right abdominal part, the 5th time on a left side strand, the 6th time at right strand, or the like.Can after the interval of one or many injection, repeat identical position.Equally, can separate drug administration by injection.Therefore, for example, a half-value dose on the same day can be used at a position, and second half is used at other position.

Perhaps, administering mode sequentially changes, as injecting weekly with successive subcutaneous, intramuscular, intravenous or intraperitoneal mode.

At 4-6 after week, in the time period of one month and a plurality of months, serve as to use at interval preferably with two weeks.Injection subsequently can be used in every month.Injection interval subsequently can be depending on patient's clinical progress and immunization therapy is replied and changed.

Can use one or more physiology's acceptable carriers or excipient to prepare the pharmaceutical composition that uses among the present invention in the mode of routine.

Can prepare, pack and indicate to be formulated in the compositions that contain the covalently or non-covalently compositions of α 2M complex comprising in the compatible carrier of pharmacy, indicate the cancer or the infectious disease that are used for the treatment of.In aspect preferred, the amount of the compositions that comprises α 2M complex that the people is used be about 1 μ g to 5mg, preferred 10 to 200 μ g, preferred 10,20,25,50,100 or 200 μ g.In preferred embodiments, comprise that the compositions of the present invention of α 2M complex is used weekly once, continue about 4-6 week, intradermal administration, site of administration sequentially changes.In preferred embodiments, the compositions of the present invention that comprises α 2M complex is used or is treated in vivo illing tissue as preliminary treatment and used after a week with 12 hours after the induced tissue necrosis.

If complex is water miscible, then it can be formulated in suitable buffer such as phosphate buffered saline (PBS) or other physiological compatibile solution.Perhaps, if the complex that obtains dissolution in aqueous solvent is poor, then it can be prepared with nonionic surfactant such as tween or Polyethylene Glycol.Thus, can with covalently or non-covalently and/or α 2M complex and physiology's acceptable solvent thing thereof be formulated as and be used for using by sucking or being blown into (through port or nose), or oral administration, through cheek, parenteral, rectal administration, or in the situation of tumor, be injected directly in the solid tumor.

In preferred embodiments, the compositions of the present invention that comprises α (2) macroglobulin comprises 9% sucrose, 5-10mM potassium phosphate in addition.In relevant preferred embodiment, the pH of compositions of the present invention is 7.

For Orally administered, pharmaceutical preparation can be liquid state, for example solution, syrup or suspension, or can be used as before using drug products with water or other suitable vehicle reconstruct.Can prepare this liquid preparation by conventional method and pharmacy acceptable additive, additive such as suspending agent (as sorbitol syrups, cellulose derivative or hydrogenation edible fat); Emulsifying agent (as lecithin or arabic gum); Non-aqueous media (as almond oil, oily ester or rectification vegetable oil); And antiseptic (as the methyl ester or the propyl ester of P-hydroxybenzoic acid or sorbic acid).Pharmaceutical composition can be tablet or the Capsule form that for example prepares by conventional method and the acceptable excipient of pharmacy, excipient such as binding agent (as pregelatinized corn starch, polyvinylpyrrolidone or hydroxypropyl emthylcellulose); Filler (as lactose, microcrystalline Cellulose or calcium hydrogen phosphate); Lubricant (as magnesium stearate, Talcum or silicon dioxide); Disintegrating agent (as potato starch or sodium starch glycolate); Or wetting agent (sodium lauryl sulphate).Can be by method as known in the art to tablet coating.

Can suitably prepare oral formulations comprises α 2M complex with sustained release compositions.This compositions can be the tablet or the lozenge form of preparation in a usual manner.

Use for suction, comprise that the compositions of α 2M complex can use suitable propellant such as dichlorodifluoromethane, Arcton 11, dichlorotetra-fluoroethane, carbon dioxide or other suitable gas form with aerosol spray from pressurized package or aerosol apparatus to use easily.In the situation of pressurised aerosol, can be by the valve decision dosage unit of the amount that discharges metering be provided.Capsule that uses in inhaler or the insufflator and medicine box such as gel can be formulated as and comprise the compositions that contains α 2M complex and the suitable powder base such as the mixture of powders of lactose or starch.

The compositions that comprises α 2M complex can be formulated as and be used for by injection as the parenteral by bolus injection or continuous infusion.Ejection preparation can exist by unit dosage form, as in ampoule or in the multi-dose container, and is added with antiseptic.Compositions can be for example form of the suspension in oiliness or aqueous medium, solution or emulsion, and can comprise batching as suspending agent, stabilizing agent and/or dispersant.Perhaps, active component can be powder type, prepares as the apirogen water through sterilization with suitable medium before using.

The compositions that comprises α 2M complex also can be formulated as rectum compositions such as suppository or delay enema, as comprises conventional suppository bases such as cocoa butter or other glyceride.

Except previous formulations, comprise that the compositions of α 2M complex also can be formulated as durative action preparation.This durative action preparation can be by implanting (for example subcutaneous or intramuscular) or using by intramuscular injection.Therefore, the compositions that for example comprises α 2M complex can for example be prepared as sl. sol. salt with suitable polymer or lyophobic dust (for example as the emulsion in acceptable oil) or ion exchange resin or as sl. sol. derivant.Liposome and emulsion are the delivery media of hydrophilic medicament or the known example of carrier.

If desired, the compositions that comprises α 2M complex can be used as can comprise one or more comprise covalently or non-covalently α 2M and the packing or the distributor of the unit dosage form of the complex of antigen molecule exist.Packing can comprise for example metal or plastic sheeting, for example blister package.Packing or the distributor instruction of can enclosing.

5.10. measurement vaccine potency

Can use the immunne response of present composition immunity experimental animal afterwards or use any immunity test as known in the art to detect the immune efficacy of the present composition by monitoring.Body fluid (antibody) is replied and/or the generation of cell mediated immunity can be used as the index of immunne response.Test animal can comprise mice, hamster, Canis familiaris L., cat, monkey, rabbit, chimpanzee etc., finally is the human agent.That vaccinated method can comprise is oral, in the brain, Intradermal, percutaneous, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal or any other immune standard way.Can be tried the immunne response of main body by the several different methods analysis; for example: by known technology such as immunosorbent adsorption test (ELISA), immunoblotting, radioimmunoprecipitation etc., or the host by the protection immunity to the anticancer or sick analysis gained of infection immune serum to interested antigenic reactivity.

As a suitable zooperal example of vaccine protective disease, can in rabbit, test the ability of vaccine-induced antibody response to antigen molecule of the present invention.Can use the adult New Zealand white rabbit of youth of male no specificity cause of disease (SPF).Every of test group is accepted the vaccine of fixed concentration.Matched group acceptance does not conform to the injection of the 1mM Tris-HCl pH 9.0 of antigen molecule.But each week or two weeks from rabbit take a blood sample and serum analysis to the antibody of antigen molecule.Can use for example existence of ELISA analysis of experiments antigen-specific antibodies.

5.11. medicine box

The present invention also is provided for implementing the medicine box of therapeutic scheme of the present invention.This medicine box is included in treatment in one or more containers or the pharmacy of prevention effective dose can be accepted the α 2M complex covalently or non-covalently of form.α 2M complex in the bottle of medicine box of the present invention can be the acceptable solution form of pharmacy, for example with the combination of Sterile Saline, glucose solution or buffer solution or the acceptable sterile liquid of other pharmacy.Perhaps, complex can be lyophilized form or dry powder form; In this case, medicine box randomly comprises the acceptable solution of pharmacy (as saline, glucose solution etc.) in addition in container, be preferably sterile solution, is used to make complex to reconstitute injection solution.

In another embodiment, medicine box of the present invention comprises pin or syringe in addition, is preferably sterile form packing, is used for the alcohol pad (packagedalcohol pad) of injection complex and/or encapsulation.Randomly comprise description, be used for clinician or patient and carry out using of α 2M complex.

6. embodiment: the inhibition of tumor and minimizing

Following result shows, can be from the serum of tumor-bearing animal purification α (2) macroglobulin complex and be used for anti-curing cancers intactly.Use the inbreeding mice as model, the immunization of the tumor cell of living being attacked subsequently with the complex of checking α (2) macroglobulin that is combined with each other with antigen molecule from tumor.Following result has supported the present invention effective especially in self-treating, comprises using from suffering from the isolating α of mammal (2) the macroglobulin complex of tumor.The immune system of the inbreeding mice of using is identical or near identical, therefore, this result can be generalized to the self-treating of treatment of cancer.

Material and method

For purification α (2) macroglobulin-antigenic molecule complexes, will dilute with 1: 1 with 0.04M Tris pH 7.6,0.15M NaCl from the serum of mice.Then mixture is applied to 65ml Sephacryl S300R (Sigma) post with same buffer balance and eluting.The 65ml post is used for about 10ml serum.Detect the positive fraction of α (2) macroglobulin and use the PD-10 post buffer to be become the 0.01M sodium phosphate buffer of pH7.5 by Dot blot.Perhaps, the 0.01M sodium phosphate buffer that can in the 65ml post, use pH7.5 as buffer to save the step of exchange buffering liquid.The fraction that will contain complex is applied to concanavalin A agarose gel post.Complex with 0.2M methyl mannose pyranoside or 5% methyl mannose pyranoside elution of bound, and to be applied to the PD-10 post be the 0.05M sodium acetate buffer of pH 6.0 to change buffer, and be applied to the equilibrated DEAE post of 0.01M sodium acetate buffer with pH 6.0.Become pure form with 0.13M sodium acetate buffer eluting α (2) macroglobulin, and by SDS-PAGE and immunoblotting analysis.

In some experiments, α 2M is available from SIGMA.BALB/c mouse derives from Jackson Labs, and (BarHarbor ME), and uses in age in 6-8 week.

In order to test the preventive means whether immunogenicity α 2M complex can form as antitumor, unless otherwise mentioned, with the isolating α 2M of 7 μ g complex to every group comprise 5 mices be used to the mouse immune of testing first.All immunity are carried out with 100 μ l PBS Intradermal.

The source that is used for the α 2M antigenic molecule complexes of immunity comprises: the serum that 1) has the mice of Intradermal 2cmMethA tumor; 2) at the serum that 50% bleach is injected directly into the mice (15 mices) after 24 hours in the 2cm tumor; 3) by the live serum of mice (15 mices) of tumor cell induction ascites of intraperitoneal injection; 4) as the mice serum that does not have tumor of negative control; 5) with the serum of the α 2M mice immunized that is compound in Meth A10 (derive from MethA tumor lysate＜peptide of 10kDa), because this complex showed the protective effect (Binder that prevents that the MethA tumor cell from attacking in the past, 2002, CancerImmunity, the same); 6) be compound in the serum of the gp96 mice immunized of Meth A10 (being called gp96-MethA10) as the use of positive control, gp96-MethA10 also showed the protective effect (Binder that prevents that the MethA tumor cell from attacking in the past, 2002, CancerImmunity, the same); 7) serum of use PBS mice immunized as negative control, is called PBS1; 8) serum of use PBS mice immunized as negative control, is called PBS2; 9) use serum from the gp96 mice immunized of liver as negative control; 10) use serum from the gp96 mice immunized of MethA as positive control because known gp96 and antigen molecule are compound and immune stimulatory (people such as Srivastava, 1986, ditto); With 11) use all through the serum of the MethA of irradiation tumor cell mice immunized, be used to detect this cell and whether can be used as positive control protective effect is provided.

Attack animal with 100,000 tumor cell Intradermal alive after one week of immunity the last time then.Measure the area of tumor.Use half radius calculation gross tumor volume (mm of meansigma methods as tumor ³).The 7th, 9.12, observed and write down observed value in 15 and 18 days.

The result

Immune result is as shown in Fig. 1 and table 2.The mice PBS 1 of negative control group and PBS 2 show that tumor development does not stop or reducing, and the decline that has shown tumor development with the animal of α (2) the macroglobulin complex immunity that derives from tumor-bearing animal at the 12nd day.At the 15th day, matched group and use were apparent from the significant difference of gross tumor volume between the animal of α (2) the macroglobulin complex immunity of tumor-bearing animal.Serum or the isolating complex of serum all effectively prophylaxis of tumours growths mice under fire from tumor-bearing animal from suffering from the ascites mice.The isolating complex of trouble mice with tumor with the bleach of induced tumor necrosis from have the tumor of being expelled to shows especially effectively protection, and observing does not have gross tumor volume to surpass 280mm ³Also find to come from α (2) the macroglobulin complex of suffering from Intradermal mice with tumor serum and more effectively suppress tumor growth than coming from the complex of suffering from the ascites mice serum.

α (2) macroglobulin that comes from normal mouse does not act on or the protective effect minimum.

Table 2

PBS

0 7 9 12 15 18

1 0 120.25 117.62 314.63 687.10 715.73

2 0 43.23 44.43 185.34 263.95 881.90

3 0 64.83 181.43 167.47 372.05 696.56

4 0 204.84 81.27 733.28 764.17 927.17

5 0 48.01 46.74 230 820.0 41136.55

Full cell

0 7 9 12 15 18

1 0 0 0 0 0 0

2 0 0 0 0 0 0

3 0 0 0 0 0 0

4 0 0 0 0 0 0

5 0 0 0 0 0 0

A2M ascites 1

0 7 9 12 15 18

1 0 249.31 328.82 0 0 0

2 0 32.251 327.10 349.99 369.56 641.11

3 0 194.52 303.58 0 0 0

4 0 201.92 183.38 199.84 353.61 492.56

5 0 140.10 156.70 0 0 0

A2M ascites 2

0 7 9 12 15 18

1 0 130.55 232.92 209.93 243.60 387.90

2 0 331.13 237.53 0 0 0

3 0 95.65 102.90 464.46 950.29 1217.38

4 0 108.85 83.56 0 0 0

5 0 91.17 128.38 132.42 234.29 350.59

A2M ascites 3

0 7 9 12 15 18

1 0 19.76 231.09 160.95 293.33 407.51

2 0 98.46 130.55 180.27 560.63 143.72

3 0 139.78 115.32 0 0 0

4 0 139.13 253.64 0 0 0

A2M bleach 1

0 7 9 12 15 18

1 0 106.66 209.07 56.80 65.02 118.79

2 0 104.77 67.60 0 0 0

3 0 64.44 87.78 25.89 0 0

4 0 217.27 211.64 0 0 0

5 0 134.00 216.39 0 0 0

A2M bleach 2

0 7 9 12 15 18

1 0 172.29 149.43 0 0 0

2 0 192.90 185.34 0 0 0

3 0 157.75 280.19 0 0 0

4 0 177.21 270.97 0 0 0

5 0 86.12 301.95 0 0 0

A2M bleach 3

0 7 9 12 15 18

1 0 24.72 82.86 180.27 0 0

2 0 6.97 0 0 0 0

3 0 60.26 42.05 0 0 0

4 0 4.38 7.70 0 0 0

A2M suffers from tumor 1

0 7 9 12 15 18

1 0 132.42 136.23 0 0 0

2 0 144.72 184.94 0 0 0

3 0 243.60 285.39 0 0 0

4 0 3.052 93.89 0 0 0

5 0 117.33 128.69 0 0 0

A2M does not suffer from tumor

0 7 9 12 15 18

1 0 149.09 255.58 331.71 507.79 605.82

2 0 103.17 73.37 65.42 0 0

3 0 123.22 203.17 346.40 527.27 605.82

4 0 117.62 297.08 119.96 158.11 220.78

5 0 101.84 129.31 249.78 382.78 448.69

From the isolating Gp96 of Meth A tumor cell

0 7 9 12 15 18

1 0 179.50 140.43 0 0 0

2 0 128.69 256.56 0 0 0

3 0 190.09 125.94 0 0 0

4 0 0127.46 0 0 0 0

5 0 148.08 91.66 85.65 350.59 523.33

With the compound 10kDa peptide of A2M

0 7 9 12 15 18

1 0 79.91 98.20 29.06 0 0

2 0 26.40 117.04 0 0 0

3 0 170.43 276.07 0 0 0

4 0 174.17 266.447 0 0 0

5 0 97.43 220.79 0 0 0

From the isolating gp96 complex of normal liver

0 7 9 12 15 18

1 0 96.41 180.27 399.59 528.85 795.92

2 0 137.19 274.02 318.56 650.14 795.92

3 0 150.80 235.68 327.67 503.19 696.56

4 0 142.40 124.12 305.77 650.14 1022.14

5 0 140.76 348.79 651.04 893.06 1149.76

With the compound 10kDa peptide of gp96

0 7 9 12 15 18

1 0 150.45 168.21 0 0 0

2 0 223.44 312.96 0 0 0

3 0 95.14 173.42 0 0 0

4 0 212.07 138.16 0 0 0

5 0 145.39 84.72 0 0 0

PBS?2

0 7 9 12 15 18

1 0 213.36 227.47 175.31 305.22 605.82

2 0 157.05 44.43 362.75 550.48 696.56

3 0 19.15 181.43 317.44 881.90 1022.14

4 0 146.39 81.27 379.61 428.46 605.82

5 0 73.16 46.74 229.73 806.35 1149.76

Conclusion

Experimental result shows that α (2) macroglobulin complex helps to treat effectively or prophylaxis of cancer.Gross tumor volume reduces to have proved that Therapeutic Method is effective.The preventive effect of before attacking with tumor cell animal being used complex is tangible.It is unexpected especially and promising that animal is used the observed significant degree of complex that comes from the animal of at first using the bleach processing that causes the tumor cell necrosis.The necrosis of tumor cell has promoted the release of tumor specific antigen molecule, its then with body fluid in α (2) macroglobulin compound.

The invention is not restricted to the scope of described specific embodiments, just as indivedual illustrations of the indivedual aspects of the present invention, the method for its function equivalent and component are within the scope of the invention for it.Except represent herein and describe, to those skilled in the art, book and accompanying drawing according to the above description, various changes in fact of the present invention are conspicuous.This change is also in additional claim scope.

All lists of references of quoting herein, patent and other open all full text are incorporated this paper into as a reference.

Claims (48)

1. treat or prevent the method for patient's cancer or infectious disease, described method comprises that the patient to described treatment of needs or prevention uses α (2) macroglobulin of amount of effective treatment or prevention patient's cancer or infectious disease and the complex of antigen molecule, and wherein said complex separates from the mammiferous body fluid of suffering from cancer or infectious disease.

2. treat or prevent the method for patient's cancer or infectious disease, it comprises step:

A) isolate the complex of α (2) macroglobulin and antigen molecule from the mammiferous body fluid of suffering from described cancer or infectious disease; With

B) use effective treatment or prevent described patient's described cancer or the described isolated complex of the amount of infectious disease.

3. claim 1 or 2 method, wherein complex is the colony with the complex of the bonded α of different antigen molecules (2) macroglobulin, wherein different antigen molecules comprise a kind of described cancer or infectious disease specific antigen being had antigenic antigen molecule.

4. claim 1 or 2 method, its be used for the treatment of cancer and wherein the patient suffer from cancer, wherein antigen molecule comes from tumor.

5. claim 1 or 2 method, its be used for the treatment of cancer and wherein the patient suffer from cancer, the effectiveness of wherein said treatment of cancer reduces by the patient tumors size or the tumor decreased number is estimated.

6. claim 1 or 2 method, it is used for prophylaxis of cancer and wherein this prevention of needs of patients, and wherein said antigen molecule shows as has antigenic antigen to described cancer specific antigen.

7. claim 1 or 2 method, it is used to keep off infection and this prevention of needs of patients, and wherein said antigen molecule shows as has antigenic antigen to described infectious disease related diseases substance specific antigen.

8. the method for claim 2, its be used for the treatment of cancer and wherein the patient suffer from cancer, and this method is included in step (b) before or simultaneously described patient is used the step of chemotherapeutics in addition.

9. the method for claim 2, its be used for the treatment of cancer and wherein the patient suffer from cancer, and this method is included in the step that step (a) is induced described mammal tumor necrosis before or simultaneously in addition.

10. the method for claim 9, wherein the step of induced tumor necrosis comprises described administration neoplasm necrosis medicine.

11. stimulate the patient to resist the method for the immunne response of cancer or infectious disease, it comprises:

A) from mammiferous body fluid isolate α (2) macroglobulin complex and

B) patient is used isolated α (2) thus macroglobulin complex immune stimulatory is replied.

12. claim 1,2 or 11 method, wherein said patient is a human patients.

13. the method for claim 11, wherein α (2) macroglobulin complex is the colony with the complex of the bonded α of different antigen molecules (2) macroglobulin, and wherein different antigen molecules comprise a kind of described cancer or infectious disease specific antigen being had antigenic antigen molecule.

14. the method for claim 11, its be used to stimulate antitumor immune response and wherein the patient suffer from cancer, this method is included in the step that step (a) is induced described mammal tumor necrosis before or simultaneously in addition.

15. the method for claim 14, wherein the step of induced tumor necrosis comprises described administration neoplasm necrosis medicine.

16. the method for prevention mammalian cancer, described method comprises α (2) macroglobulin of using effective dose and the complex of antigen molecule, and described complex separates from described mammiferous body fluid, and described mammal suffers from precancerous lesion or polyp.

17. claim 1,2,11 or 16 method, wherein mammal is behaved.

18. prevent the method for first mammalian cancer, described method comprises α (2) macroglobulin of using effective dose and the complex of antigen molecule, described complex separates from the second mammiferous body fluid, and described second mammal suffers from precancerous lesion or polyp.

19. the method for claim 16 or 18, wherein complex comprises the multiple complex of the bonded α of different antigen molecules with one or more (2) macroglobulin, and wherein different antigen molecules comprise at least a described cancer or infectious disease specific antigen being had antigenic antigen molecule.

20. claim 1,2,11,16 or 18 method, wherein said complex are described patient from body.

21. claim 1,2,11,16 or 18 method, wherein said complex before using through purification.

22. claim 1,2,11,16 or 18 method, wherein said body fluid is the vascular fluid.

23. the method for claim 22, wherein the vascular fluid is the serum that comes autoblood.

24. claim 1,2,11,16 or 18 method, wherein body fluid is outer ascites or cerebrospinal fluid of vascular.

25. pharmaceutical composition, it comprises (a) from the isolated multiple α of the mammiferous body fluid of suffering from cancer or infectious disease (2) macroglobulin-antigenic molecule complexes, and described multiple complex comprises at least a α of comprising (2) macroglobulin and shows as the complex that tumor or infective agent specific antigen is had antigenic antigen molecule; (b) pharmaceutical carrier of effective dose.

26. the pharmaceutical composition of claim 25, wherein said body fluid are the vascular fluid.

27. the pharmaceutical composition of claim 26, wherein said vascular fluid is the serum that comes autoblood.

28. the pharmaceutical composition of claim 25, wherein said body fluid are outer ascites or cerebrospinal fluid of vascular.

29. vaccine, it comprises: (a) from the isolated multiple α of the mammiferous body fluid of suffering from precancerous lesion or polyp (2) macroglobulin-antigenic molecule complexes, described multiple complex comprises at least a α of comprising (2) macroglobulin and shows as the complex that tumor specific antigen is had antigenic antigen molecule; (b) pharmaceutical carrier of effective dose.

30. the vaccine of claim 29, wherein multiple α (2) macroglobulin-antigenic molecule complexes is through purification.

31. pharmaceutical composition, it comprises: (a) comprise α (2) macroglobulin and the complex with tumor or the antigenic antigen molecule of infectious disease; (b) be selected from following medicine: HSP-peptide complexes, antineoplastic agent, antibody, cytokine, antiviral agents, antifungal agent, antibiotic and chemotherapeutics; (c) pharmaceutical carrier of effective dose.

32. the pharmaceutical composition of claim 31, its Chinese medicine are chemotherapeutics.

33. the pharmaceutical composition of claim 31, wherein α (2) macroglobulin-antigenic molecule complexes is through purification.

34. comprise the method for the complex amount of α (2) macroglobulin and antigen molecule in the increase mammalian body fluid, described method comprises induces described mammiferous neoplasm necrosis.

35. the method for claim 34, wherein the step of induced tumor necrosis comprises administration neoplasm necrosis medicine.

36. medicine box, it comprises α covalently or non-covalently (2) macroglobulin-antigenic molecule complexes that pharmacy can be accepted the treatment of form or prevent effective dose in one or more containers.

37. comprise the method for the complex amount of α (2) macroglobulin and antigen molecule in the increase mammalian body fluid, described method comprises induces described mammiferous neoplasm necrosis.

38. preparation comprises α (2) macroglobulin and has the method for the complex of precancerous lesion, tumor or the antigenic antigen molecule of infectious disease that it comprises:

A) extract serum from the patient who suffers from precancerous lesion, tumor or infectious disease; With

B) reclaim α (2) macroglobulin-antigenic molecule complexes from described serum.

39. the method for claim 38, it is used for the treatment of or prophylaxis of cancer, and described method comprises the step of recovery complex of described patient being used the amount of effective treatment or prophylaxis of cancer in addition.

40. the method for claim 38, wherein the step from described serum recovery α (2) macroglobulin-antigenic molecule complexes comprises:

A) solid phase that comprises α (2) macroglobulin binding molecule being contacted with serum enough makes α (2) macroglobulin-antigenic molecule complexes be incorporated into the time of solid phase;

B) remove the material that is not incorporated into solid phase; With

C) from α (2) macroglobulin-antigenic molecule complexes of solid phase elution of bound.

41. the method for claim 39, wherein α (2) macroglobulin binding molecule is α (a 2) macroglobulin specific antibody.

42. the method for claim 39, wherein α (2) macroglobulin binding molecule is the part binding fragment of CD91.

43. preparation comprises α (2) macroglobulin and has the method for the complex of precancerous lesion, tumor or the antigenic antigen molecule of infectious disease that it comprises:

A) extract serum from the patient who suffers from precancerous lesion, tumor or infectious disease; With

B) fractionated serum is with enriching of alpha (2) macroglobulin-antigenic molecule complexes.

44. the method for claim 38 or 43, it comprises the step that serum is contacted with the medicine that promotes formation covalent bond between α (2) macroglobulin and the antigen molecule in addition.

45. the method for claim 44, its Chinese medicine are protease, ammonia, methylamine or ethamine.

46. the method for claim 38 or 43, wherein said body fluid are the vascular fluid.

47. the method for claim 46, wherein the vascular fluid is the serum that comes autoblood.

48. the method for claim 38 or 43, wherein body fluid is outer ascites or cerebrospinal fluid of vascular.

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