CN1269013A

CN1269013A - Moleculars withch give out multiple active parts

Info

Publication number: CN1269013A
Application number: CN98804085A
Authority: CN
Inventors: 乔治·怀特塞兹; 詹姆斯·B·塔兰勃姆; 约翰·格里芬; 马塞·马蒙
Original assignee: Dean And Colleagues Of University Of Kazakhstan; Advanced Medicine Inc
Current assignee: Dean And Colleagues Of University Of Kazakhstan; Innoviva Inc
Priority date: 1997-04-11
Filing date: 1998-04-09
Publication date: 2000-10-04
Also published as: EP0973551A2; KR20010006280A; AU7106998A; JP2002503223A; AU743028B2; BR9808521A; WO1998046270A2; WO1998046270A3; CA2286692A1

Abstract

Pharmaceutical compositions for polyvalently presenting an agent for therapy are described. The pharmaceutical compositions contain a polyvalent presenter. In one embodiment, the polyvalent presenter has a formula as follows: (Y)-(X-A)n, wherein Y is a framework, X is a direct bond or a linker, A is a presented functional group, and n is greater than ten and is an integer selected such that the presented groups can interact with a plurality of target binding sites. The composition also can include a pharmaceutically acceptable carrier. Alternatively, the presenter itself can serve as its own pharmaceutically acceptable carrier. Methods for treating diseases or conditions also are described. The methods involve administering to a subject a plurality of groups A such that the treatment occurs. The treatment occurs by the interaction of a polyvalent presenter with a plurality of target binding sites B. Other aspects of the invention include polyvalent presenters packaged with instructions for use in the aforementioned methods and methods for designing polyvalent presenters which are useful within the methods of this invention. The polyvalent presenters disclosed herein provide for specificity in binding, which has a number of advantages. Furthermore, the polyvalent presenters permit positive and negative interactions.

Description

Present the molecule governmental fund of a plurality of active parts

This working portion is subsidized by NIH Grants GM30367 and GM39589. Subsidized by NIH Grant 1-S10-RR04870-01 and NSF Grant CHE88-14019 at the NMR of Harvard facility. Therefore, government enjoys certain right to the present invention. Related application

The application require priority be U.S. Provisional Application number 60/043,781 (applying date: on April 11st, 1997) and 60/043,826 (applying date: on April 14th, 1997), this two incorporated by reference in the lump at this. The application also with the application that is entitled as " multivalence is presented thing associating storehouse and application thereof ", U.S. Provisional Application number 60/043,288 (the applyings date: on April 11st, 1997), with the application that is entitled as " multivalence is presented thing associating storehouse and application thereof ", the U.S. fixes tentatively application number 60/043, the 918 (applying date: on April 15th, 1997) relevant.

Background of invention

Multivalence acts on ubiquity in the biology, and it is very important to the function of biosystem. The multivalence effect is by many binding events to each ligand-receptor (for example) simultaneous interactions (seeing Fig. 1) between the species of two separation. The interaction of part and acceptor runs through whole biology. Part is included in the molecule of presenting information in the biosystem or being subjected to the protein effect. The kind of part has, for example, and medicine, hormone, signaling molecule, cell surface marker, toxin, substrate, biological regulator, neurotransmitter and lymphokine. Acceptor comprises with the part effect and receives the molecule of information from part. Most acceptors are protein and comprise protein receptor, antibody and enzyme. Some acceptors are nucleic acid and comprise DNA and the regulatory region of RNA.

Many medicines are and the interactional part of single receptor (some medicines are the acceptors with single ligand interaction). In biology many important events from a plurality of parts and a plurality of acceptor in the effect. The multivalence effect spreads all over biology. The multivalence effect generally is to occur in cell surface and/or is many relating in groups or the interaction of cluster acceptor. It also is important that multivalence acts on the macromolecule interaction aspect that relates to simultaneous multiple spot adhesion.

People explore the concept of multivalence effect. In the research on mechanism article of attempting to provide the adhesion of influenza virus and cell surface, effect is inquired into to multivalence. (people such as Whitesides, " medical chemistry magazine " (J.Med.Chem.), 1995,38,4179-4190; The people such as Sigal, " U.S. chemical institute magazine " (J.Am.Chem Soc.), 1996,118,16,3789-3800). The people such as Kiessling (" chemistry and biology " (Chemistry and Biology), 1996, Vol.3, No.2,71-77) have created and have produced the structure model that the multivalence carbohydrate shows to study and adjust the bio-identification event. The people such as Kiessling think that these models can be used as " be beneficial to and explore the interactional chemical tools of multivalence albumen-glucosides ". Be entitled as " where we go thus? " a joint of list of references in the people such as Kiessling illustrate " except can also be used as the probe of biological function to exploring the useful multivalence carbohydrate of biometric discrimination method ". (FEBS LETTERS 1989,252,1,1-4) the multivalence inhibitor that suppresses the microorganism adhesion is used in suggestion to Matrosovich. Matrosovich illustrates " some principles in can utilizing in order to make this multivalence structure people ' medicine be delivery system ' planning known; for example, the unit price in many duplicate is suppressed bioactive molecule and the biocompatible polymer of solubility or particulate carrier coupling ". Matrosovich also illustrates " correctness of this practical application near antimicrobial design and explore and can estimate in the future ... ".

Summary of the invention

Today, " high-affinity selectivity binding events has been arranged the most of thought about receptor-ligand binding " (see the above-mentioned article of Kiessling the 71st page). Usually, the attempt of optimization receptor-ligand binding has concentrated on specifically part and specifically binding ability or the selectivity aspect of receptor binding site in the independent binding events. For example, part is that choosing is based upon the interaction on its known good combination capability foundation discussion, perhaps avoid using the part that is considered to weak combination, perhaps before choosing is used, weak binding partner is carried out chemical modification to improve its binding ability. Needn't avoid weak binding partner because it is the part that multivalence of the present invention is presented thing.

The present invention is based on, the observation of at least part of multivalence effect that occurs to receptor-ligand binding with unconventional comprehensive method based on us, this mode based on the multicomponent multivalence is presented thing how with the interactional understanding of the set of target binding site. Our research is different from than the usual manner of observing this interaction (as the receptor-ligand binding of separating) on the isolated foundation. The unconventional comprehensive method of receptor-ligand binding and multivalence effect makes us recognize that the multivalence effect can be used as the basis of appropriate design medicine in this observation biosystem, for example, initial basis, even be widely used aspect the many various disease for the treatment of or the indication. The unconventional universal way of this observation receptor-ligand binding even let us are recognized based on choosing of the particular ligand of independent binding ability and are needed not to be most important parameter in the design multivalence medicine.

It is to make up and arrange many identical or different group A (such as part) by the framework (such as polymer frame) of presenting thing in the multivalence that consists for the treatment of disease or indication to form that multivalence of the present invention is presented thing. The many types of binding site of curative effect and single ligand binding wherein, and cover with the many types of binding site of many types of ligand binding and obtain be by in patient body, present with the multivalence of adhesion that thing is gathered target binding site B or target binding site B array (for example, the receptor binding site of bunchiness on cell surface), group A makes up and arranges and can carry out according to result for the treatment of at framework. It is that multivalence is presented thing and is integrated into the result who has an effect on the interface that is consistent with target binding site B that target binding site B set is covered. It is the result that multivalence is presented thing and many set of target binding site B occurred conformation interfacial interaction that target binding site B array is covered. Cover being not to be limited to continuous obstacle, we blanket a question discussing in detail in below " detailed Description Of The Invention ".

In order to describe, we will discuss in detail and cover effect, present thing because it relates to the multivalence with polymer frame. As shown in Figure 2, the multivalence with polymer frame 1 is presented thing P and can be supposed a configuration that is consistent with interface 3, and for example, the surface comprises the set in a target set site 5 owing to its interaction. The multivalence that conforms to of one or more adhesions is presented thing and has basically been covered a target set site B array (being comprised of an above target binding site B set), and this array improves or provides multivalence to present the result for the treatment of of thing. Covering can be that physics is covered, and for example, presents thing physics with multivalence and covers target binding site B, and/or be that solid is covered, and for example, a plurality of group A that presented thing by multivalence cover a plurality of target binding site B. Cover needing not to be obstacle continuous or that closely meet, but can be the discontinuous or loose obstacle that meets, as shown in Figure 2, and cover also and needn't cover the target binding site fully. In some instances, covering layer will be the glue-line that is consistent with the surface of containing target binding site B array.

The present invention even relate to the method for the treatment of disease or indication. The method comprises and uses some group A to the patient so that because target binding site B set or array is covered and disease or indication are produced therapeutic action in the patient body in an instantiation. Covering of target binding site B set in the patient body presented thing by single multivalence be integrated into generation interaction realization on the interface that is consistent with target binding site B. Covering of target binding site B array in the patient body presented thing by many multivalence be integrated into generation interaction realization on the interface that is consistent with target binding site B. The method also relates to the part that the multivalence that meets some standard is presented thing or special selection is arranged in other instantiations, such as group A, framework or connector, the multivalence use of presenting thing.

The invention still further relates to the pharmaceutical composition that is delivery of therapeutic agents by the multivalence effect. This pharmaceutical composition contains multivalence presents thing. This multivalence is presented thing and be can be represented by the formula:

(Y)-(X-A) _n

Wherein, Y is framework, and X is straight key or connector, the functional group that A is presented, and also each A can be identical or different, and n is the integer greater than 10. Select such to present thing so that the group A that is presented can interact with target binding site B set. Said composition also can contain pharmaceutically suitable carrier, and perhaps, this presents thing itself as pharmaceutically suitable carrier. To present thing be to form like this to multivalence in an instantiation, namely select n and allow-(X-A) part adheres on the Y, meet with the interface phase that contains target binding site B set so that multivalence is presented thing, and cover these target binding sites B set by being used for the patient. X is connector in another instantiation, and it is independently part rather than the part of Y or A. N is greater than 10 and be integer. Select such present thing so that when being used for the patient multivalence present thing and gather with target binding site B and be consistent. Y is aggregation framework in another instantiation, and the functional group that A is presented, X are linking groups, and n is greater than 10 and be integer. Select such structure so that when being used for the patient multivalence present thing and gather with target binding site B and be consistent.

The present invention relates to the method for adjusting adhesion effect between a plurality of surface-binding biomolecules and a plurality of the second molecule on the other hand. The method comprises that making a plurality of surface-binding biomolecules and multivalence present thing is combined. This multivalence is presented the framework that thing contains a plurality of the third molecules of adhering. The third molecule is combined with the part of a plurality of surface-binding biomolecules by the third molecule of while entropy ground raising and will be adjusted adhesion effect between surface-binding biomolecules and the second molecule less than the three-dimensional blocking-up of a part of surface of being combined with the third molecule-binding biomolecules by framework.

The present invention is said method on the other hand, and wherein to present thing be non-glucide to multivalence and have an appointment greater than 10 enhancement factorβ.

The present invention is that following multivalence is presented thing pAA (Gal-β) on the other hand, pAA (Gal-α), pBMA (Gal-β) or pBMA (Gal-α).

The present invention is the method that prevents ricin and red blood cell adhesion on the other hand. The method comprise with ricin and effective dose be selected from pAA (Gal-β), pAA (Gal-α), the multivalence of pBMA (Gal-β) or pBMA (Gal-α) is presented the thing contact.

The present invention is that preparation is selected from the method that the multivalence of pAA (Gal-β) and pBMA (Gal-β) is presented thing on the other hand. The method comprises the O-L with Gal-β₁NH ₂With poly-(N-acryloxy succinimide) or poly-(butadiene-common maleic anhydride) reaction, then cancellation reaction.

The present invention is that preparation is selected from the method that the multivalence of pAA (Gal-α) and pBMA (Gal-α) is presented thing on the other hand. The method comprises the C-L with Gal-α₂NH ₂With poly-(N-acryloxy succinimide) or poly-(butadiene-common maleic anhydride) reaction, then cancellation reaction.

The present invention is that pAA (GlcNAc-β) multivalence is presented thing on the other hand.

The present invention is the method that suppresses patient's internal fertilization on the other hand, comprises that using effective dose pAA (GlcNAc-β) multivalence to the patient presents thing.

The present invention prepares pAA (GlcNAc-β) multivalence to present the method for thing. The method comprises β-L with GlcNAc-₁NH ₂With poly-(N-acryloxy succinimide) reaction, then cancellation reaction.

The diagram summary

Fig. 1 is the flow chart of describing unit price and two kinds of reactions of multivalence.

Fig. 2 is that the multivalence of describing polymerization is presented the diagram that thing " is covered " binding site set and target binding site B array.

Fig. 3 is the diagram of describing ricin-cell interaction.

What Fig. 4 described is the general plan of inducing sperm acrosome reaction.

Fig. 5 describes multivalence to present the synthetic flow chart of thing galactoside.

That Fig. 6 a describes is the multivalence galactoside cohesion inhibition activity of polymerization and the anti-RCA of Gal of polymer₁₂₀The relation curve of molar fraction.

That Fig. 6 b describes is the multivalence galactoside cohesion inhibition activity of polymerization and the anti-RCA of Gal of polymer₆₀The relation curve of molar fraction.

Fig. 7 describes the N-acetyl glucosamine of multivalence polymerization and the diagram of galactoside structure.

Fig. 8 is the diagram of describing by unit price and multivalence GlcNAc inducing mouse sperm acrosome reaction.

Fig. 9 a is the diagram of describing by the multivalence GlcNAc inducing mouse sperm acrosome reaction of polymerization.

Fig. 9 b is the figure that described with the carrying out of pAA (GlcNAc) gained the sperm (%) of acrosome reaction.

Figure 10 is the curve map of describing multivalence GlcNAc Inhibit sperm by polymerization-ovum combination.

Figure 11 a describes the schematic diagram that pMVMA (NeuAc) generates.

Figure 11 b describes pMVMA (NeuAc; R) schematic diagram that generates.

Figure 11 c describes the schematic diagram that pAA (Gal) generates.

Figure 11 d describes the schematic diagram that pBMA (Gal) generates.

Figure 12 a describes pAa (SLe^X) schematic diagram that generates.

Figure 12 b describes pAA (bacitracin; R) schematic diagram that generates.

The detailed Description Of The Invention drug design

The present invention relates to design the method that multivalence is presented thing, the wherein said thing of presenting is for giving patient treatment disease or indication. The method comprises that making up and arrange 10 above group A forms the multivalence for the treatment of disease or indication and present thing on framework. This structure and arrangement are based on the designed thing of presenting and provide the ability of result for the treatment of owing to covering target binding site B set. Target set site B array in a preferred embodiment in the patient body is presented thing by one or more multivalence and is covered. Multivalence is presented thing and target binding site B and is integrated into interaction on the interface that is consistent or some multivalence and presents thing and cause this phenomenon of covering with the interaction that target binding site B is integrated on the interface that is consistent.

Term used herein " multivalence is presented thing " will be described in detail hereinafter. Multivalence is presented thing and is comprised having 11 or the ten more than one multicomponent molecules of presenting group A (having in conjunction with 11 or 11 above target binding site B abilities) that are connected on the framework. In brief, multivalence is presented thing and be can be represented by the formula:

Y-(A) _nWherein, " Y " represents framework; The functional group that " A " representative is presented; And " n " representative is greater than 10 integer, it so that the group of being presented can interact with many binding site B.

Term " disease or indication " comprises with multivalence of the present invention presents thing, by the multivalence effect, such as group A and the interaction that contains the multivalence acceptor of target binding site B, the disease for the treatment of or indication. Become the disease of a part of the present invention or indication generally can binding site B and group C between interaction be feature. B described here and C are multivalence, and wherein, " B " represents a plurality of binding sites, and " C " representative in essence relevant with disease or indication and also with the interactional part of B or group. Group C can be identical with group A in some instances, and it can be presented thing by multivalence of the present invention and present, thereby cause a plurality of (C) and a plurality of (B) competition when interacting with binding site B. Group C can be different from group A in some other example. When binding site B was natural generation, group C both can be that what to synthesize also can be natural. For example, C can be natural sugar, peptide, protein (for example, the part of cell surface receptor such as viral antigen), or synthetic (group that for example, can present by silicon implant or prosthese). Group C can be from the teeth outwards or in solution, but must be that multivalence is presented. Term " disease or indication " also will comprise and be related to by identification or be subjected to natural multivalent molecule, for example, suppresses the α of influenza virus and target cell interaction₂Huge glomus cell molecule, the symptom of impact or disease. If bioprocess is very complicated, then those of ordinary skills can present the thing effect by the test multivalence specific site (being that purpose is to disturb multivalence to present the concrete event of thing) whether with a plurality of (C) and a plurality of (B) between interaction relevantly be familiar with disease or whether indication comprises the multivalence interaction. Described in detail in disease and symptom will be below " method for the treatment of disease or the indication " chapter.

Term " patient " comprises suspects to suffer from or to have suffered from the disease that will treat or treat or the mammal of indication. Therefore, it is human that the present invention is used to treatment, domestic animal, and domestic animal, zoo animal etc., for example, people, ox, cat, dog, sheep and mouse. The present invention is used to design the medicine for the treatment of human patients in preferred embodiment.

Term " treatment disease or indication " comprises that presenting thing with multivalence of the present invention treats or prevent these diseases or indication to the disease suffering from needs and treat or the patient of indication. Term " treatment " comprises the patient is produced beneficial effect, for example can significantly alleviate at least symptom or the performance of disease or indication. Therefore, the arbitrary measure " treatment " refers to from least a symptom of improving disease or indication to the continuous process of cure diseases or symptom. Treatment comprises that also using alone multivalence to present thing or present thing with other multivalence is combined with, even is combined with other therapeutic agent that is not multivalence is presented thing.

Term " makes up and arranges some group A at framework " and comprises that operating multivalence presents the various components of thing to generate the thing of presenting that can realize treating disease or indication function. This operation can realize with the basis of the viewpoint of comprehensive method interaction at group A and target binding site B set. This operation comprises the location, and large minispread and selection multivalence are presented each composition of thing, for example, selects multivalence to present the framework of thing, attachment and group A. For example, this term is will comprise group A and the framework location of position relative to each other, perhaps with respect to being used for that group A is connected to the optional connection molecule on the framework or being used for the location of the key attachment of monomer on the connecting frame. This term also will comprise group A, the selection of the particular type of framework and/or attachment.

When framework is that the multivalence ability that its selection will form according to it particular type of " covering " target binding site B set when presenting thing a part of decides. For example, some frameworks can form the colloid substance reason barrier that covers binding site B set. Framework can also be presented in the thing the required different qualities that has in the multivalence that will design according to it and select. For example, can according to " flexibility " of framework and/or, even after connecting group A, framework is given the ability that multivalence presents thing with flexibility and is come Selection Framework.

Can also give the ability that multivalence presents thing with needed feature according to group A and select group A. The binding ability of group A is the factor in its selection course, and the present invention thinks that partly weak conjugated group A can be used in its multivalence forming process, emphasizes that this point is very important at least.

The location of group A on framework also is that " make up and arrange " multivalence is presented the part of thing. The location can for example, utilize multivalence environment molecular model known or prediction, realizes on the steric basis of target site B set. The location can be carried out along several disalignments with respect to framework. For example, target site B can arrange with the distance of Average distance 10 dusts with adjacent target binding site B, and therefore, group A can be along framework in the horizontal direction arrangement or the location that are suitable near adjacent target binding site B. Should be appreciated that group A needn't locate for the distance of catering between the adjacent target binding site, and must be to be suitable for being close to the principle location. When at framework location group A, except considering distance factor, also to consider other factors, such as flexibility and the profile that comprises the interface of target binding site of attachment. For example, pliable and tough attachment can play regulating action in vivo, like this, even the distance between the group A is not in full conformity with the distance between the adjacent target binding site B, perhaps when the profile curves at interface and when unsmooth, also can be near the target binding site. The degree of depth of binding pocket (nest), for example, the 2-20 dust also can be known or allow group A for example, can to position to predict with the concrete length of attachment along the axle that is basically perpendicular to framework.

" make up and arrange " and also will comprise type and the length of selecting group A is connected to the attachment on the framework. Can select according to the ability that group A is presented to the target binding site B in the binding pocket length of attachment, wherein, the degree of depth of binding pocket and/or diameter are known or predictable. The chemical property of attachment, for example, hydrophobicity or hydrophily also can be according to selecting about the knowledge of target binding site surrounding environment, for example, this attachment may through known be hydrophobic or hydrophilic passage or environment arrival destination. Can also be according to attachment with required character, such as flexibility, give the ability that multivalence presents thing and select.

Term " result for the treatment of " is to comprise that multivalence presents the ability that thing is realized its predictive role such as treatment or prevent disease or indication. Multivalence is presented the result for the treatment of of thing can be with measuring with the art-recognized determination method that designed multivalence be presented thing treatment disease specific or indication, the determination method that perhaps will carry out with art-recognized technology and/or described herein but do not belong to above-mentioned all kinds of determination method and measure. Present thing and meet and impel its ability of being combined with target binding site B set multivalence if effect comes from multivalence, then " owing to covering effect " will consider result for the treatment of.

Term " covers " that the physics that comprises target binding site B is covered or physics covers and solid is covered, and for example, the three-dimensional inaccessible and/or raising part A loci B of target site B occupies. The example that physics is covered comprises in the binding site B set formation of gluey layer or barrier, can prevent that like this group except multivalence is presented group on the thing is near this site. Described physical barriers can be pasta strip barrier shown in Figure 2. In both cases, the formation of gluey layer or barrier or pasta strip barrier all be by multivalence present in the group on the thing at least some with corresponding target site on the binding partners specific binding and the biologic specificity real estate dried rhizome of rehmannia.

Target binding site B is referred to that by " solid is covered " the group A that binding site B is connected on the framework surrounds in velcro shape mode. A group A is incorporated into binding site, and other group A can three-dimensionally block identical binding site B or adjacent target binding site.

Term " target binding site B set " comprises with single multivalence and presents the binding site B that the thing molecule is consistent. For example, if a multivalence is presented thing 10 group A that are connected on the framework are arranged, then the set of these binding sites will be the span of the binding site B that meets with this molecule, for example, in fact 10-20 binding site although may only have two binding sites to be occupied. Should be noted that not every binding site B has an effect with group A, so some site may not be occupied.

Term " target binding site B array " comprises the set of an above target binding site B on the interface. Binding site B in the array needn't have equal space or position, but can put at random on all directions along the axle that with respect to the interface is different directions, for example, and according to factors such as the profile at interface and binding site colony configurations. For example, the profile at interface can be level and smooth or crooked, also can be mobile.

Term " interfacial interaction is consistent " comprises that multivalence presents thing and be consistent at the interface with the configuration that is consistent, for example, can be near the molecular domains of group A, the interaction that occurs with target binding site B set. This interface may be, but not limited to,, the surface, but to comprise interactional border. It is to comprise this situation that term " is consistent ", namely with regard to the surface in whole or in part with regard to, it is closely or firmly that multivalence is presented interface between thing and the surface. Under latter event multivalence present thing can with surface some site close proximity and be connected not tight with the other parts on surface. Term " interaction " is to comprise that the positivity and the negativity that describe in detail below interact. Dependence is enough to the flexibility of adjustment interface profile, and multivalence of the present invention is presented thing and some interfaces, and for example, cell surface is consistent. This being consistent can be meeting along the span of the target binding site B of some. This span preferably is at least about 50, is at least about 100, is at least about 1000, upward to being at least about 10⁶Individual target binding site B. The intermediate range of above-mentioned value for example, is at least about 50-1000, or is at least about 1000-10⁶, also be a part of the present invention. For example, also comprise stride values scope as the combination of any above-mentioned value of the upper limit and/or lower limit. Above-mentioned scope also will comprise site and the interior site of target binding site B array in the target binding site B set.

Term " interaction " is to comprise negative interaction, such as inhibition, and for example influenza, and positive interaction, as make the interaction of signal conduction, or take two surfaces to together interaction. For example, multivalence present thing can with, for example, lip-deep target binding site B set combination, and signal or inducing cell consumed energy in the transfer cell. The example of this repercussion effect comprises apoptosis (programmed cell death), and the clone expands, for example, and B cell or T cellular replication, cell migration, such as the movement of cell to some direction, the release of shla molecule, for example, hormone, such as insulin, cell factor such as I12; Prostaglandin, encytosis or pinocytosis or exocytosis, the active transport of some materials (inwardly or outwards), electroactive inducing action, the perhaps differentiation of special cells or dedifferente effect. Present thing with multivalence the adjusting of all these effects or processing are all belonged to the scope of the invention. Interaction is not anti-stick company in some instantiations.

Term " steric stabilization effect " refers to that such mechanism, multivalence presents thing and can three-dimensionally suppress by being combined in specific manner with one of them surface biological approaching of two surfaces. It is different from the static stabilization of realizing in the colloidal mixture by adding water-swelling polymer, adds polymer and is beneficial to keeps colloidal particle in solution. The non-stable surface of wish that is incorporated into of molecule in this case single-mindedly. For example, referring to " the electronic three-dimensional stabilization of polymeric colloid (the Electrosteric stabilization of polymer colloids) " of A.Sung and I.Piirma, Langmuir, 1994,10,1393-8; U.Genz, B. D ' Aguanno, " structure of steric stabilization colloid (the Structure of sterically stabilized colloids) " of J.Mewis and R.Klein, Langmuir, 1994,10,2206-12. This concept also is different from transplants polymer to colloid or surface of liposome. The steric stabilization effect is to adhere with surperficial covalency that will be stable by molecule to realize under latter event. Polyethylene glycol is the prototype that exemplifies. For example, referring to K.Zhulina, " transplanting polymer to the theory (Theory of steric stabilization of colloid dispersions by grafted polymers) of colloidal dispersion steric stabilization effect " of O.Borisov and V. Priamitsyn, " colloid and interface are learned " (Colloid and Interfacial Science), 137:495-511,1990.

Term " multivalence is presented thing " includes the multicomponent molecule of being presented group A more than 11 or 11, and multivalence is presented thing and can be combined with corresponding target binding site B, and group A adheres on the framework. Multivalence of the present invention is presented thing can use following representation:

(Y)-A _n

Multivalence of the present invention is presented thing and can be used following representation in some instantiation:

(Y)-(X-A) _n

" Y " represents framework in above-mentioned expression formula; The functional group that " A " representative is presented; The optional linking group of " X " representative, this group is used for group A is adhered to framework Y; And n representative is greater than 10 integer, the group and a plurality of binding site B interaction selecting like this n to make to be presented.

The selection of Integer n is presented fully many group A, and then reaches the purpose for the treatment of disease or indication. Select Integer n that a plurality of group A are presented by multivalence. Integer n is preferably greater than 10 to about 10 greater than 10⁶, more preferably 50 to about 10⁶, about 100 to about 10⁶, or about 1000 to about 10⁶ N between institute's train value also belongs to the scope of the invention, and is for example about 100 to 1000 greater than 10 to about 1000 greater than 10 to about 100, and 1000 to 100,000. For example, comprise that also the combination as any above-mentioned value of the upper limit and/or lower limit all belongs to the scope of n.

Term " multivalence is presented thing " or " multivalence mode " they are art-recognized and comprise A, the multivalence demonstration of B or C, like this, and a plurality of A, B or C functional group are different from their unit price equivalent. For example, many A can show positive cooperativity or negative cooperativity or the non-synergitic biological effect of single A. In addition, multivalence is presented thing and can be acted synergistically independently, and this effect will be illustrated by the β factor of explaining more in detail hereinafter. Multivalence is presented thing or multivalence mode and is comprised that multivalence presents the purposes of thing, and it is different from slowly-releasing compound well known in the prior art or medicine is presented system. Multivalence is presented thing and is designed to the multivalence functionalization and does not discharge the material of monoradical in order to the pharmacological activity of regulating drug. In addition, owing to discharge and the diffusion monoradical, the compound of slowly-releasing generally all acts on the site that is different from the administration site, presents thing for multivalence, can do like this. Framework

Term " framework " (Y) comprises supporting construction or the skeleton of being made above about 10,000 material by molecular weight, also can be presented by multivalence above a plurality of group A can adhere to. Adhesion can be adopted and allow the group mode that multivalence shows on framework realize. Group A has just been adhered on the framework before giving patient's administration in some instances. Group A is used to the patient in other example, then is pooled in vivo on the framework, for example, forms multivalence of the present invention by compiling of self and presents thing. Frame part must have enough mean hydrodynamic radius striding across the distance between the adjacent acceptor, and 100 dusts will be arranged or more than 100 dusts. Such appearance and size could allow a plurality of functional group A adhere on the framework to be attached to simultaneously the target acceptor, for example, and cell surface receptor. Those of ordinary skill will recognize that all the mean hydrodynamic radius of polymer can obtain with the rough calculation of canonical statistics mechanics method.

" skeleton " at some instantiation middle frames can also comprise attachment (claiming later on the skeleton attachment), and they link together the monomeric unit of framework. Concrete skeleton attachment will discuss in detail in the chapters and sections about polymer frame below. The skeleton attachment is dissociable in some instances, and for example, the skeleton attachment is that facile hydrolysis changes.

Being used in multivalence, to present framework in the thing be can be with group A adhesion thereon and can multivalence present the sort of framework of group A. The framework that is used for purpose in the body both had been applicable to the vivo medicine-feeding mode, also was applicable to binding mode in the body. The example of several types framework includes, but not limited to polymer, liposome, protomere, colloid, dendrimer and biologic grain. These types will simply be discussed below, then further discuss in detail in the chapters and sections about covalency and non-covalent framework. Only just having in the beginning of covalency and non-covalent framework discussion about the detailed description of each in these frameworks, so only is the restriction that does not conveniently consist of framework in order to discuss. Should be appreciated that the present invention will comprise all types of frameworks of can multivalence presenting above-mentioned group A.

Term " polymer " " or " polymerization " be art-recognized, and comprise the structural framing that contains the repeated monomer unit. If the framework of opinion qualification polymerization also must multivalence be presented " A " group, could treat disease or indication like this. This term also comprises copolymer and homopolymer, for example, and synthetic or natural copolymer and homopolymer. Straight chain polymer is in branch polymer and cross-linked polymer are also included within. Polymer can not be glucan in preferred embodiment.

Term " liposome ", " protomere " and " colloid " is art-recognized. These terms also comprise its derivative, such as liposome derivative, crosslinked liposome etc.

Term " dendrimer " is art-recognized. Dendrimer includes the specific subclass of the branch polymer in many generations. Every generation has all been created a plurality of branch points in dendrimer. Can not be dendrimer at the preferred embodiment middle frame.

Framework of the present invention can contain " biologic grain " in some instances. Term " biologic grain " comprises covalent molecule, for example, and sugar, protein, lipid, little molecule, protein aggregation body and nucleic acid, and non-covalent particle, for example, modified cell (as being derivatized, chemical modification or infected by exogenous nucleic acid) or modified virus, for example, virion. " biologic grain " that uses as framework is different from the particle of native state, presents functional group A because described framework is modified multivalence. The covalency framework

Monomeric unit at an instantiation middle frame can be by covalently bound. Representational covalency framework comprises crosslinked liposome, and biologic grain (such as sugar, protein, peptide, lipid, or little molecule), and polymer (for example, referring to Siraganian, the people such as R.P., " immunochemistry " (Immunochem.), 1975,12,149-155; Wofsy, the people such as C., " IMMUNOLOGY KEY WORDS INDEX (J.Immunol.), 1978,121,593-601; Barlocco, the people such as D., " (pharmacy " (Farmaco.), 1993,48,387-96; Castagnino, the people such as H.E., " Japanese heart journal " (Jpn.Heart J.), 1990,31,845-55; Costa, the people such as T., " biochemistry pharmacy " (Biochem.Pharmacol.), 1985,34,25-30; Dembo, the people such as M., " IMMUNOLOGY KEY WORDS INDEX (J.Immunol.), 1979,122,518-28; Hlliger, the people such as P., " Proc. Natl. Acad. Sci.USA (Proc.Natl.Acad.Sci. U.S.A.) ", 1993,90,6444-8; Piergentili, the people such as A., " pharmacy " (Farmaco.), 1994,49,83-7; Portoghese, the people such as P.S., " medical chemistry magazine " (J.Med. Chem.), 1991,34,1292-6; Kizuka, H. and Hanson, R.N., " U.S. chemical institute magazine " (J.Am.Chem.Soc.), 1987,30,722-6).

Protein such as albumin can be used as presenting system (Roy, the R. of a large amount of groups in some instances; Lafemere, C.A., " Canadian Journal of Chemistry " (Can.J.Chem.), 1990,68,2045-2054); This is a kind of imitated natural sugar albumen.

Polymer is used as the framework that multivalence is presented thing in other example. Polymer is general frame system (people such as Spaltenstein, " american chemical magazine (J.AM.Chem.) ", 1991,113:686; The people such as Mammen, " medical chemistry magazine (J.Med.Chem.) ", 1995,38:4179). Group A of the present invention is adhered on the framework in preferred embodiment, and this framework comprises by the polymeric skeleton that connects genomic constitution. Reactive polymer can be used in the present invention's " framework " who describes in detail below in some instances.

Polymer can prepare with means known in the art (Sandler, S.R.; Karo, W., " polymer synthesizes (Polymer Syntheses) "; Harcourt Brace:Boston, 1994; Shalaby, W.; Ikada, Y.; Langer, R.; Williams, J., " biology and biomedical significant polymer (Polymers of Biological and Biomedical Significance (ACS Symposium Series 540) ", American Chemical Society:Washington, DC, 1994). Polymer design must be able to be had good pliability; The length of the attachment between the distance of biologically active side chain and polymer backbone and the group can be designed and control.

Polymer frame has many advantages. They can be designed to minimum inappropriate chain a plurality of group A are attached on a plurality of group B binding sites simultaneously. The multivalence of polymerization is presented thing can be easy to, be synthesized with assembling rapidly and (people such as Spaltenstein, " U.S. chemical institute magazine (J.AM.Chem.Soc.) ", 1991,113:686; The people such as Mammen, " medical chemistry magazine (J.Med.Chem.) ", 1995,38:4179). Polymer also allows presenting the various physical propertys of thing, for example, and the flexibility that conforms to; Dissolubility; Hydrophily is adjusted, and regulates uniformity and flexibility in the solution of different temperatures and ionic strength.

The high molecular polymer chemistry is very flourishing subject, and organic polymer is presented for multivalence a very important compounds is provided. These compounds have very high molecular weight and can present very a large amount of groups that copies; They can present an above group simultaneously; Their the biomembranous transmission of passing generally is confined, and therefore, their life-spans in special compartment can be controlled in vivo. Aggregation framework provides the various large molecules that are easy to synthesize, and near the biologically active scope.

In preferred embodiment, be used for modified polymeric material of the present invention and have low antigenicity and hypotoxicity. The optional next and water compatible of aggregation framework in preferred embodiment, aggregation framework can have various molecular weight and have the various different groups that are adhered to polymer backbone. Polymer backbone of the present invention obtains at an easy rate by synthetic.

The inherent biocompatible polymer that contains the functional group that is suitable for adding side chain is preferred (Shalaby, W.; Ikada, Y.; Langer, R.; Williams, J., " biology and biomedical significant polymer (Polymers of Biological and Biomedical Significance (ACS Symposium Series 540) ", American Chemical Society:Washington, DC, 1994). The example of polymer comprises PEO or polyethylene glycol (Harris, J.M., " PEG chemistry: biotechnology and biomedical applications (Poly (ethylene glycol) Chemistry:Biotechnical and Biomedical Applications) ", Plenum:New York, 1992; Horton, D., " carbohydrate chemistry and biochemical development (Advances in Carbohydrate Chemistry and Biochemistry) ", Academic Press:San Diego, 1995), and, for example, the derivative of acrylamide and NVP, continuous oligomeric glycol oligomer, continuous glucan oligomer etc.

Other is used for preferred framework of the present invention and has certified purposes, for example, as plasma expander, drug excipient or adhesive, food additives, or as the inertia or the erodable material that are used in the body. For example, can use PEG, poly-(lactic acid), poly-(glycolic acid) and PVP.

Present the synthetic preferred polymers of thing for multivalence and contain reactive group such as the carboxylic acid of activation. A large amount of synthetic polymer with natural contain carboxylic acid function group or can be enough carboxyl acid modified, and they are used in the body. Such polymer can form with the group A that quilt is presented covalently bound, for example, and amido link. The carboxylic acid function group of containing inner cyclisation is desirable especially such as the polymer of acid anhydrides or succinimide group. Other preferred polymer comprises the inferior unit of deriving from maleic anhydride or malic acid. The example of copolymer comprises phenylethylene-maleic anhydride and alpha-olefin-maleic acid (such as divinyl ether-maleic acid). In other example, can use sodium carboxymethylcellulose, chondroitin sulfate and poly-(methacrylate/acrylate) material. In other examples, there is not activating carboxy acid's polymer can use, for example dextran sulfate yet.

Other aggregation framework that is worth exemplifying comprises: poly-(ester), and poly-(acid anhydrides), poly-(carbohydrate), how pure, poly-(acrylate), poly-(methacrylate), poly-(ether) and poly-(amino acid).

Other aggregation framework that are worth exemplifying comprise: poly-(glutamic acid), poly-(aspartic acid), glucan, dextran sulfate, poly-(maleic anhydride-altogether-vinyl ethers), poly-(succinimide), poly-(acrylic anhydride), PEG, poly-(lactic acid), poly-(glycolic acid), PVP, poly-(phenylethylene-maleic anhydride), α-maleic acid, hyaluronic acid, sodium carboxymethylcellulose, chondroitin sulfate, poly-(acrylate), poly-(acrylamide), poly-(glycerine) and starch.

Those of ordinary skill should be appreciated that any polymeric material that can present a plurality of group A all may be applicable to the present invention. Can carry out modification to polymer, for example, as mentioned above, or by derivatization, as use difunctional cross-linking reagent, to obtain being suitable for adhering and to present the functional group (will describe in detail hereinafter) of group A. Copolymer may be preferred in some cases.

Table 1 polymer for example

PEG poly-(acid amides) poly-(peptide), poly-(amino acid) poly-(aspartic acid), poly-(glutamic acid) poly-(lysine), other oroteins (gelatin) poly-(ester) poly-(lactic acid), poly-(tartrate) glycan fiber element of polyactide poly-(glycolide) poly-(caprolactone); Alginates; Starch, glucan derivative gathers (NVP)	Poly-(vinyl acetate-vinyl esters) poly-(acrylamide) poly-(urethane) poly-(methacrylate) poly-(acrylate) poly-(maleic acid) poly-(acid anhydrides) poly-(ortho esters)
		Copolymer and degradable attachment encapsulate in order to protect in order to reach target with the group that will adhere

The skeleton attachment

Should comprise the coupling part in some instances, namely between each monomeric unit of skeleton " attachment ". The example of " skeleton attachment " can comprise carbohydrate, carbamate, and acid amides, ether, thioesters, thioether, carbonic ester is connected with ester. The skeleton part can connect with the attachment of cleavable in some instances. The multivalence material life-span in vivo may depend in part on its molecular dimension. By between middle-sized oligomer, placing linking group and by these groups of control stability in vivo, can controlling the polymer life-span in vivo. Therefore, the purposes of degradable attachment in polymeric skeleton of non-functionalization can be with helping remove polymer. Generally, the attachment of cleavable is different from the attachment of connecting framework and group A. This cleavable connects the segment that will cause the polymerizable functional that formation is less, and this segment will be so small as to the degree that is enough to clean kidney. The degradable attachment can comprise that for example, very easily the attachment of hydrolysis comprises ester, carbonic ester or oxalates group.

The framework that the multivalence of studying is in some instances presented thing can contain dendrimer. Dendrimer is this area approval, and it comprises the high branched polymer of a series of low-molecular-weights, for example, usually is monodispersed. The advantage of dendrimer is that it has very clear and definite molecular structure, and their solution phase viscosity for their molecular weight is quite little. With respect to straight chain polymer, they can present group in the mode of compact package, can be conducive to like this arrive at the acceptor that clusters round on cell surface. The quantity of group A can be controlled among the dendrimer exactly, and group interior distance, uniformity and the rigidity that can regulate the dendrimer molecule. The glycan class may just belong to a kind of dendrimer (people such as Sabesan, " U.S. chemical institute magazine ", 1992,14:8636). The begin chain group of dendrimer can be used as framework (Roy in an instantiation, the people such as R, " based on the synthetic and antigenic property (Synthesis and Antigenic Properties of Sialic Acid-Based Dendrimers) of sialic dendrimer ", Acs Symp.Ser 1994). Non-covalent framework

A plurality of group A can also be connected on the non-covalent framework. The example of non-covalent framework comprises liposome, protomere, colloid, protein aggregation body, modified cell and modified virus particle. For example, group A can be strapped in the head group of liposome molecule, film or surface upper (Kingery-Wood, the people such as J.E., " U.S. chemical institute magazine ", 1992,114,7303-5; Spevak, the people such as W., " U.S. chemical institute magazine ", 1993,115,1146-7; Spevak, the people such as W., " U.S. chemical institute magazine ", 1996,39,1018-20).

Liposome and protomere are art-recognized, and comprise the macroscopic particle that is formed by the lipid aggregation, for example, and surfactant. Multivalence is presented thing and can be utilized liposome or protomere to present group (Spevak, the people such as W., " U.S. chemical institute magazine ", 1993,115,1146-7 in an instantiation; Charych, the people such as D., " chemistry and biology (Chem.﹠ Biol.) ", 1996,3,113-120). This system is the shape of imitating target cell tightly, and can be designed to present the surface tightly meeting target cell aspect types of radicals and the density. For example, contain the lipid molecule of group A, for example, can again be built into liposome as the neuraminic acid (NeuAc) of polar head group. Liposome has gratifying biocompatibility, and is synthesized quite easily. In addition, liposome can be designed to have the sensor effect. The liposome of polymerization can be used to feel the metamorphosis of liposome, shows this variation by the absorption that shows the inner chromophore of UV/ (ratio) film (for example, crosslinked poly-diacetylene). For example, virus can detect by gamut (Lan Zhihong) with the combination of liposome, and this point has been art-recognized.

In other example, comprise that the biologic grain of modified cell for example or modified virus can be used as the framework that group A multivalence is presented. Therefore, protein, peptide, the glycan class, the intact cell (such as red blood cell) of cell membrane segment or modification, the bacterial cell of modification or the virus of modification can be used as framework in some instances. The frame part of activation

Term used herein " framework of activation " refers to the said frame part, comprises covalency and non-covalent frame part, its contain can by " activated group " activation functional group, then with at least one functional group, auxiliary group and/or interval radical reaction. Suitable functional group comprises, for example, and carboxyl (form of acid or salt), hydroxyl, sulfydryl, acid amides, carbamate, amino, ketone, aldehyde, alkene, fragrance (hydrocarbon) etc. Can polymer with before functional group contacts with polymer activation (pre-activation), or in the presence of functional group activated polymer (activating then and there).

Activation step can comprise with can experience and the group of nucleophile or electrophilic substance reaction polymer is carried out derivatization. In addition, activated polymer makes them can lean on cation, anion or gene triggering mechanism participate in the bipolarity addition reaction (such as 1,3-and 1, the addition reaction of 4-bipolarity), ring adds reaction (such as the reaction of Diels-Alder type) and polymerization physical reaction, and these all belong to the scope of the invention.

By using, for example, annular or straight chain acid anhydrides, the ester of activation is (such as N-hydroxy-succinamide, nitrophenol, 4-hydroxyl-3-nitrobenzene-sulfonic acid etc.), acyl chlorides, imidazolidine (as, derive from carbonyl dimidazoles), carboxylic acid and ester can with activated carboxylic, be used for the reaction with nucleophile. Also can pass through at carboxyl and reagent, such as dicyclohexyl carbodiimide, 1-(3-dimethylamino-propyl)-3-ethyl carbodiimides, alkyl chloroformate, chlorosilane, pyridinium salt and Bu₃N etc., between form adduct and activate the polymer that contains carboxylic acid.

The choosing of group that is suitable for the activation of carboxyl function group is apparent for those of ordinary skills. Reaction system ought to or require activation then and there or pre-activation, and this point also should be apparent for those of ordinary skills.

Can use with, for example, halo alkyl formate or acyl ester (such as isobutyl chlorocarbonate, p-nitrophenyl chloro-formate etc.), the carbonate activation hydroxyl that cyanogen bromide or phosgene reaction form. Containing the oxidation reaction of using the periodate compound to carry out aspect the polymer (such as glucan and other glycan class) of vicinal diols group can be used to obtain reactive carbonyl moiety at polymeric skeleton in utilization. Activation is apparent with the addition method of the polymer of hydroxyl for those of ordinary skills.

Can be with two thiobis pyridinyl compounds with the polymer of sulfydryl, such as 2,2 '-two thiobis (5-nitropyridine), 2,2 '-two thiobis (pyridine) etc., activation. Being used for activation is apparent with the addition method of the polymer of sulfydryl for those of ordinary skills.

It should be appreciated by those skilled in the art that above-mentioned priming reaction only is used for illustrating, also have many versions that are suitable for existing procedure.

In case aggregation framework is activated, it just can with at least one functional group, auxiliary group or interval group, or their mixture reaction. Perhaps, the polymer of activation can with functional group, the reaction of the combining form of auxiliary group and/or interval group. The functional group part

The functional group that multivalence prepared according to the methods of the invention is presented thing partly comprises can adhere to frame part, on auxiliary group and/or the interval group, and can to present the mode of function, such as in order to treat disease or indication, carry out the group that multivalence is presented. The functional group part can be identical or different. Term " a plurality of functional group part " or " a plurality of radicals R^3”Refer to surpass an above functional group part, each functional group in these groups is identical or different independently of one another. These functional groups can be identical or different in the classification of type of functional group, such as, a functional group can be carbohydrate, and another functional group can be antibiotic. These functional groups can be different functional groups in similar functional group also, and for example, functional group is two kinds of different carbohydrate. The functional group of being presented in " the homopolymer type is presented thing " situation can be identical, and the functional group of being presented in " the heteropolymer type is presented thing " situation just can be different, for example, and from variety classes or with the different groups in the kind. In order to describe conveniently hereinafter with in claims name R³ ₁To R³ _nRepresent numerous R³In different members.

It is as herein described that to present the functional group that thing presents by multivalence of the present invention be functionalization. When functional group is adhered to multivalence and is presented on the frame part of thing functional group just with they function contribution out. As mentioned above and reaffirm that here this point is different from art-recognized medicine and presents system, for example, polymeric drug is presented system, and this system discharges therapeutic agent, and wherein therapeutic agent provides its function with the form that discharges. For example, functional group for example, directly provides therapeutic efficiency by be used for providing its function with (that is, a plurality of) binding site B more than 11 or 11 is biological single-mindedly in preferred embodiment of the present invention. Presenting thing when multivalence in other example is that for example, when the heteropolymer type was presented thing, some functional group was (such as R³ _n) just may be not and site B effect, provide other function and replace. For example, functional group may affect R³ ₁With the interaction of binding site B, therefore, what it rose is the function of reinforcing agent. Functional group is (such as R in other example³ _n) can be complementary functional group (such as auxiliary group), it can affect the physical characteristic that multivalence is presented thing, as passes through dissolubility or the ability of cell membrane. In other example, can utilize functional group to follow the trail of the present invention and present thing, for example, as the detectable marker of presenting thing (such as fluorescence or radioactive labels).

Functional group of the present invention part both can be that what to synthesize also can be natural. In addition, functional group part can be classified according to physical characteristic such as molecular size, namely functional group can have low, in or HMW. The example of natural function group's composition includes, but not limited to natural carbohydrate, protein (such as IgE or erythropoietin(EPO)), peptide and other known drug. The example of synthetic functional group composition includes, but not limited to the Counterfeit Item of peptide, with combinatorial chemistry technique or the synthetic functional group of rational drug design technology. The particular type of the functional group part that can use according to the inventive method will be entitled as below in the chapter of " type of functional group " and discuss in detail. Group A

The group A that multivalence is presented thing comprises can be to present the mode of function, for example, in order to treat disease or indication, carries out that multivalence is presented and at the group that for example adheres on the framework on the framework. Group A can be identical, also can be different. Term " a plurality of group A " refers to more than one group A, and each the group A in these " a plurality of " is independent of each other identical or different. These functional groups can be identical or different in the classification of type of functional group, such as, a functional group can be carbohydrate, and another functional group can be antibiotics. These functional groups can be different functional groups in similar functional group also, and for example, functional group is two kinds of different carbohydrate. The group A that is presented in " the homopolymer type is presented thing " situation can be identical, and the group A that is presented in " the heteropolymer type is presented thing " situation just can be different, for example, and from variety classes or with the different groups in the kind. In order to describe conveniently hereinafter with in claims name A₁To A_nRepresent the different members among the group A.

It is as herein described that to present " the group A " that thing presents by the present invention be functionalization. Group A also provides their function when group A is adhered to multivalence and present on the frame part of thing. As mentioned above and reaffirm that here this point is different from art-recognized medicine and presents system, for example, polymeric drug is presented system, and this system discharges therapeutic agent, and wherein therapeutic agent provides its function with the form that discharges. For example, group A for example, directly provides therapeutic efficiency by be used for providing its function with (that is, a plurality of) binding site B more than 11 or 11 is biological single-mindedly in preferred embodiment. In some instances, for example, presenting thing when multivalence is heteropolymer type when presenting thing, and some group is (such as A_n) just may be not and site B effect, provide other function and replace. For example, A_nMay affect A₁With the interaction of binding site B, therefore, what it rose is the function of reinforcing agent. A in other example_nCan be the auxiliary group of functionalization, it can affect the physical characteristic that multivalence is presented thing, as passes through dissolubility or the ability of cell membrane. In other example, can utilize group A to follow the trail of the present invention and present thing, for example, as the detectable marker of presenting thing (such as fluorescence or radioactive labels).

Group A of the present invention both can be that what to synthesize also can be natural. In addition, group A can classify according to physical characteristic such as molecular size, namely group A can have low, in or HMW. The example of natural group A includes, but not limited to natural carbohydrate, protein (such as IgE or erythropoietin(EPO)), peptide and other known drug. Synthetic group A includes, but not limited to the Counterfeit Item of peptide, with combinatorial chemistry technique or the synthetic group of rational drug design technology. Group A does not choose from following groups in preferred embodiment: carbohydrate, polymyxins such as PB, the compound that beta-lactam, furans are derived, the compound that pyrans is derived and antibody are selected separately or are united use with any other member in these groups no matter is. Group A is not derivative or the segment of above-mentioned group A in other examples. The particular type of group A will be below " type of group A " one the joint in discuss in detail. Each presents the quantity of group A in the thing

The various physical propertys of framework can affect multivalence and present the ability that thing is adjusted group A and binding site B interaction. Can change the usefulness that multivalence is presented thing by the quantity that changes the group equivalent, namely increase or reduce usefulness. For example, a large amount of sticky points can make the skeleton of presenting thing be retracted on the surface, binding site B is displayed on this surface, and three-dimensional stability usefulness reduces, but aspect the competitiveness blocking-up of target site and/or the excitability of biological response/Antagonism aspect usefulness increase to some extent.

System research with substituting group effect on the polyacrylamide polymers parent of 0.2 equivalent sialic acid (SA) is shown, the substituent net charge of skeleton, size and hydrophobicity all can affect by presenting thing the success (people such as Mammen of biological function, " medical chemistry magazine ", 1995,38:4179). In a word, change the substituent person's character of skeleton and will affect compatibility and steric stabilization effect. And the usefulness of polymerization inhibitor can reduce along with the increase of substituting group electric charge and size. Coulomb and cubic phase mutual effect are more stretched chain, and the usefulness of steric stabilization effect aspect is reduced. Present a plurality of not isoplastic heteropolymer types and present thing

The heteropolymer type is presented and can be used to provide than larger intensity and the selectivity of equivalent unit price interaction. For example, the group by presenting another kind of type is (such as A₂) interactional sum is increased, then interactional overall strength also can increase. By regulating respectively the Gene A that will present₁And A₂Quantity can increase interactional selectivity. For example, with group A₁And A₂The thing of presenting can be than only with group A₁Or A₂Present thing more closely with (site) B₁And B₂Effect.

Except using different group A (they give unique pharmaceutical properties to the described thing of presenting), the use of a plurality of different group A can be protected the infringement that is not subjected to various different pathogens.

For example, multivalence is presented thing and can be presented the group A with receptor acting on the pathogen surface in an instantiation₁With with the group A of the second pathogen effect₂ At another middle B that gives an actual example₁And B₂On identical pathogen. The type of group A

Group A of the present invention comprise when presenting in the multivalence mode, can be used for treating disease or indication group. These groups can be known group or medicines, or studied the multivalence that relates to disease specific or indication interact after selected new group or medicine. The present invention also provides purposes (Kiessling, the L.L. of above-mentioned known group A; Pohl, N.1., " chemistry and biology ", 1996,3,71-7) and the integrated approach of the new group of exploitation, and the usefulness of presenting to improve the known monomers group by the multivalence form. Should be realized that and since easier present some application of thing near these multivalence of action site may be more catchy than alternate manner. Should be appreciated that unapproachable situation can change by more directly giving the action site administration.

Notified in some instances and relate to cell-pathogen and interact, cell-cell interaction, pathogen-extracellular matrix interacts and the interactional group of pathogen-pathogen can the multivalence mode be presented thing in the present invention and presented.

For example, described in appended embodiment and appendix, a group that exemplifies is N-acetylneuraminic acid (a kind of sialic acid), and it is the natural binding site of influenza agglutinin. Interaction between this sugar and its lecithin is that influenza virus adheres to the first basic step on its target cell. Overwhelming majority multivalence sugar can become group A, comprise Neu5Ac (2,6) galactolipin, Neu5Ac (2,6) lactose, NeuAc (a2,3) Gal (b1,3) GalNAc, heparin sulfate and cerebroside, they are adhered at different virions and have shown important effect aspect the host cell.

The known group that can nurse one's health other disease that relates to the multivalence effect or indication can be used to the present invention and present thing in other example. For example, can select can the adjustment for the treatment of thrombosis time the interactional group A of multivalence between the blood platelet.

Unit price form administration with them when the present invention presents thing and can be included in treatment disease or indication in preferred embodiment has shown known drug or the compound that does not have positive effect. In these examples, when carrying out multivalence according to the present invention when presenting, these medicines have become significant effective again. Therefore, the invention provides some existing medicines and non-pharmaceutical group, they can be used as group A and are impregnated in the present invention and present thing. It will be understood by those skilled in the art that group A can have more than one biological effects, and/or can be used for the treatment of more than one diseases or indication. Be also to be understood that some group A will be used to treatment and relate to the interactional disease of multivalence or indication and/or some group A and will be used to treatment and relate to the interactional disease of non-above-mentioned multivalence or indication. Below used term " medicine " refer to the possible group A that is easy to discuss.

For example, the invention provides the medicine powerful to central nervous system. 9-tetrahydroaminoacrine or donepezil can be treated Alzheimer disease as group A. Available abstinyl is done group A treatment alcohol addiction disease in another example. Treat acute and/or chronic ache and/or inflammation can be used anodyne (for example, acetaminophen, aspirin, brufen, naproxol, pentazocine, indocin or diflunisal) and arcotic (such as Ropivacaine or Remifentanil) make group A. Treatment pain and/or anesthetic dependence syndrome can be used, for example, dihydrocodeinone, the fragrant meperidine of the third oxygen, Dilauid, morphine, amidone or dihydrohydroxycodeinone are made group A. The present invention also provides narcotic purposes, and these anesthetic comprise, for example, and adrenaline, Kerocaine, Carbocainum, methohexital, Bupivacaine and procaine. Multivalence of the present invention is presented thing can also make group A with anticholinesterase, for example, and pyridostigmine or neostigmine. Also hypnotic can be mixed the present invention and present thing, for example, available fluazepam, amobarbital, triazolam, temazepam or quinalbarbitone. Group A also can be antitussive, for example, and pseudoephedrine or codeine. The present invention also makes group A with antimigraine, and for example, ergot derivative (such as ergotamine or desernil) or sometheptene, or serotonin (5-HT) antagonist are such as sumitriptan. Group A can also be the therapeutic agent of movement disorder, such as Bonamine or hyoscine.

Group A can be muscle relaxant also in other is used, such as pyridostigmine, and neostigmine, Scoline, Mivacron, rocky ammonium, Rocuronium, Vecuronium, dantrolene, prohe tatriene, baclofen, Chlorzoxazone, methocarbamol or papaverine. It is available to feel sick, for example, and prochlorperazine, chlorpromazine, trimethobenzamide, perphenazine, hydroxyzine or Ondansetron treatment. Parasympatholytic also can be used as group A, such as akineton, and phenobarbital, ergotamine, bentyl, hyoscyamine, glycopyrrolate. In addition, parasympathomimetics, such as Romotal, pilocarpinum, urecholine is risen the happiness dragon, and yogimbine also can use. Group A can also comprise the Antiparkinsonian medicine, example hydrochloric acid benzhexol, bentropine, kemadrin, levodopa, bromocriptine, carbidopa, amantadine. The present invention also provides psychotropic drugs to present purposes in the thing in the present invention. In addition, anxiolytic, such as L0, buspirone, librium, peacefulness, chlordiazepoxide, stable and ALprazolanic also can be mixed the present invention and present in the thing. Antidepressants, such as nardil, parnitene, parozetine, fluoxetine, Sertraline, amitriptyline, nortriptyline, imipramine or protriptyline also can use. Antipsychotic drug, such as Clozapine, prochlorperazine, haloperole, loxapine, thioridazine, fluphenazinum, profit is sent ketone, mesoridazine, triperazine, Olanzapine, or chlorpromazine also can be used as group A. Can also use psychoanaleptic, such as azoxodone or ritalin. Sedative, such as mebaral, quinalbarbitone or temazepam also can be used as group A. Can use with, for example, non-ammonia ester, Gabapentin, phenytoinum naticum, 3-mesatoina, ethotoin, Lamotrigine, the first amber, the benzene amber, the second amber, cabamozepine, the multivalence of phenacemide or cabamazepine is presented thing treatment epileptic attack. Can also use antisympathetic, such as phentolamine. Group A can also comprise, for example, anticonvulsive drug (such as Fosphenytoin), antidepressants (such as Mirtazapine), can be used to treat the group of multiple sclerosis (such as glatiramer) or epilepsy (such as Topiramate), or can be used to suppress the group of Angiogenesis (such as the vasoinhibitor of generally acknowledging).

The present invention also provides the purposes to the powerful group A of cardiovascular system. For example, adrenergic is disliked piperazine, Terazosin, prazosin, methyldopa ethylester, clonidine and labetalol such as quinoline. Hypertensin conversion enzyme inhibitor in another example, such as captopril, lisinopril, tradolapril or enalapril also can use. Multivalence is presented the thing of presenting that thing can also be made into to present angiotensin ii receptor antagonist such as Losartan or Valsartan. The present invention also provides antiarrhymic, such as norpace (disopyramide), and procainamide, quinindium, propafenone, flecainide, Tocainide, PR, sotolol, amiodarone or digoxin. β-blocking agent in other example, such as Timolol, metoprolol and atenolol also can be used as group A. Calcium channel blocker also can be as the group of being presented in another example. The example comprises nifedipine, nicardipine, CRD401, Felodipine and verapamil.

Diuretics, such as acetazolamide, ethacrynic acid, furosemide, antisterone, amiloride, chlorothiazide and hydrochlorothiazide also can be used as group A. Vasodilator, such as papaverine, hydralazine and amrinone also can be used for the present invention and present thing. The present invention also provides the purposes of vasopressor, such as aramine, and phenylephrine or isoprel. Group A can comprise in another example, for example, and mecamylamine. Group A can be hypolipidemic, such as clofibrate, and gemfibrozil, the parent cuts down his spit of fland, Lovastatin, atorvastatin or nicotinic acid. Can use, for example, danaparoid makes group A and treats the formation of deep layer vein blood vessel. Available antihypertensive in another example is such as gutron.

A comprises for treatment cancer group, for example, antiandrogen (such as Leuprorelin or flutamide), cytocide is (such as adriamycin, adriamycin (doxorubicin), taxol, endoxan, busulfan, neoplatin or α 2-interferon), antiestrogen (such as TAM) and antimetabolite (such as fluorouracil, methotrexate (MTX), mercaptopurine or thioguanine). Group A also comprises hormone, such as 6 MAPs, and estradiol, Leuprorelin, first ground hydroxyprogesterone, Sandostatin LAR Depot or growth hormone-release inhibiting factor. Group A comprises in another example, for example, and Irinotecan, gemcitabine, toptecan, nilandrone or docetaxel.

The present invention presents thing and can be used for gastrointestinal applications in other example. For example, anti-spasmodics or anticholinergic drug, such as bentyl, hyoscyamine, glycopyrrolate, benzhexol hydrochloride or atropine also can be used as group A. Anorectic also can be used as group A, such as dextro-amphetamine, and Benzphetamine, Phentermine or ormazindole. Group A can comprise antidiarrheal agent in other example, such as loperamide, and Diphenoxylate or Sandostatin LAR Depot. The present invention presents thing and can also comprise, for example, and proton pump inhibitor (such as Takepron or Ao Mola azoles) or vasodilator (such as opioid).

The group A that regulates internal system also can multivalence present. For example, contraceptive, such as ethinodiol, ethinyloestradiol, norethindrone, norquen mestranol, Desogestrel or 6 α-first-17-OH progesterone. The group A of conditioning diabetes also can use, for example, and glibenclamide or chlorpropamide. The anabolism medicine such as Testolactone or stanozolol, also can be mixed the present invention and be presented in the thing. Androgen, such as methyltestosterone, testosterone or FL also can be used as group A. Antidiuretic such as minirin, also can be used as group A. The present invention presents thing and also can the multivalence mode show, for example, and the calcitonin effect. Can also use estrogen, such as diethylstilbestrol. Group A also can be glucocorticoid, such as triamcinolone or betamethasone. The present invention also provides and uses progestogens, for example, norethindrone, Etynodiol, left-handed 18-methylnorethindron and ethinyloestradiol are as group A. Also use in another example thyroid drug (such as triiodothyronine or levothyrocine) or antithyroid drug (such as methimazole). Available in other example, for example, Cabergoline is too much sick as group A treatment blood prolactin. Available in another example, for example, miglitol or insulin lispro treatment diabetes. The present invention also provides the hormone depressant, such as the purposes of danazol or Coserelin. Group A comprises oxytocic in other example, such as methergine or oxytocins. Prostaglandin also can be used as group A, such as mioprostol, and PGE₁Or PGE₂。

Multivalence of the present invention is presented thing can also be used for dermatologic application. There is the example of the group A of dermatology effect to comprise, for example, anti-acne drug (such as 2-13-Cis-retinoic Acid, Adapalene or retin-A). Other can be used for dermatological applications, for example, the treatment itch group A comprise, for example, other beclomethasone, benzocainum, hydroxyzine, Fluticasone, Mo Mitasong, fluocinolone acetonide, Clobetasol or desoximetasone.

Group A can also act on the clot that condenses. For example, poly-(A) can contain anti-coagulants or heparin. Group A can comprise in other example, and for example, Antithrombin III or platelet suppressant drug are such as Abciximab and/or ticlopidine.

In other example, can select group A to the influence power of immunoregulation effect according to group A. For example, histamine interacts from the multivalence that the release of mammary glandular cell and basocyte relates between cell surface anaphylactogen and the IgE acceptor. Group A can comprise in this case, for example, antihistamine, such as diphenhydramine, Loratadine, brompheniramine, anarexol, fenazil, RMI 9918, fexofenadine, tall and erect benzyl phthalazines and/or meclastine. Group A comprises that mammary glandular cell stablizes medicine in another example, such as Lodoxamide and/or Cromoglycic acid.

Other is regulated immune group A and comprises, for example, and steroids (such as triamcinolone, Beclomethasone, cortisone, dexamethasone, prednisolone, methylprednisolone, Beclomethasone, or Clobetasol), histamine H₂Antagonist (such as famotidine, cimetidine or ranitidine), immunosuppressive drug (such as imuran or cyclosporin). The group that anti-inflammatory activity is arranged, such as Su Lingda, etodolac, ketoprofen or ketorolac also can be used as group A. Also available antihistamine is such as fexofenadine or tall and erect benzyl phthalazines.

The group A that regulates respiratory system also can use. For example, mucolytic/expectorant (such as guaiacol glycerol ether), anti-inflammatory agent (such as Cromoglycic acid, 9-removes the fluorine fluocinolone acetonide, beclomethasone or budesonide) also can be presented thing for the present invention. The present invention also provides the purposes of bronchodilators as group A, and it comprises, for example, and Ipratropium Bromured, alotec, terbutaline, neoisuprel, albuterol or theophylline. The medicament for the treatment of asthma also can use, such as zafirlukast or Zileuton.

Group A also comprises antibiotic, such as quinolone (such as Ciprofloxacin, Norfloxacin, Ofloxacin, Yi Nuosha or Lomefloxacin), sulfonamide (such as salicylazosulfapyridine or sulfamethoxazole) or, for example, PB, bacitracin, neomycin, terramycin, TOB, sulfacetamide, phosphonomycin and penicillin, antiviral agent (such as trifluorothymidine, Zidovudine (AZT), zalcitabine (ddV), didanosine (ddl) or stavudine (d4T), or other RTI, to sell such as Merck, protease inhibitors (is exerted Buddhist such as the thiophene quinoline, indinavir or ritonavir), acyclovir, famciclovir, ICN-1229, NVP, cidofovir or penciclovir), and antiparasitic (such as thiabendazole, chloroquine, fanasil, pyrimethamine, Mefloquine, flagyl, albendazole or ivermectin), or antifungal (such as butenafine, Butoconazole, terconazole, Tioconazole, clotrimazole or MCZ). Infection such as ophthalmia with special antibiotic therapy special site is preferred under certain conditions, and this point is obvious to those skilled in the art.

Penicillin assistant agent such as benemid also can be used as group A in other example.

The urethra cartridge bag that can be used as group A is drawn together, for example, and ethacrynic acid medicine such as sulfinpyrazone. Antibiotic, such as carindacillin, furantoin, acidum nalidixicum, neomycin, bacitracin or PB also can be used as group A. The present invention presents thing also can present anti-spasmodics (example hydrochloric acid oxybutynin or flavoxate). The calcinm oxalate calculus preventive medicine also can be used as group A, for example, and allopurinol. Also can use the hypertrophy of the prostate change factor (such as Terazosin or Finasteride). The present invention presents thing can also be used for the treatment of cystitis, for example, makes group A with pentosan.

The present invention presents thing and also is used for treatment for ocular inflammation. For example, β-blocking agent (such as brominide or betoxolol), anti-inflammatory agent (such as clopatadine), or be used for the treatment of glaucomatous medicine (as drawing smooth prostate) and all can be used as group A. The present invention also provides the purposes of carbonic anhydrase inhibitor, for example, uses daranide, and methazolamide or Dorzolamide are made group A.

For dental applications, for example, canker sore, available, for example, Amlexanox is made group A.

Group A can be independently selected from erythropoietin in other example, fujimycin 506, HGH is organized the blood plasma enzyme activation factor, platelet activating factor, the interleukin stimulating factor, granulocyte stimulating factor, macrophage stimulation factor, target are the little molecule sercretogogue of protein acceptor, such as the growth stimulation sercretogogue, EPO sercretogogue and insulin secretagogue.

Some instantiations (only being illustrative rather than definitive thereof) that multivalence is presented thing comprise the multivalence sialic acid of HA on the resisiting influenza virus, Sialyl Le^XWith two derivatives that the blood platelet that contains CD62P forms, the anti-multivalence Sialyl Le that contains the endothelium of E and P selection albumen^X, in conjunction with the GABA that contains two different acceptor sites_BThe barbiturate of coreceptor (Benzodiazepine) is in conjunction with leukocytic RGD and Sialyl Le with integrin and selection albumen^X, reach the C in conjunction with tumour (it combines gal-x-gal) and complement cascade protein_5AAnd Gal-X-Gal. Auxiliary group

Term used herein " auxiliary group " refers to change multivalence and presents the group part that thing and/or multivalence are presented thing part (for example, frame part, functional group part, interval group etc.) characteristic. The character that can change comprises that for example, dissolubility is (at water, fat, lipid, in the biological fluid etc.), hydrophobicity, hydrophily, the flexibility of framework, antigenicity, molecular size, molecular wt, distribute biocompatibility in the Half-life in vivo, body, immunogenicity, stability is attached to intensity on the multivalence target etc.

Those skilled in the art understand, although be not that all it is overlapping each other basically that these characteristics have many, and auxiliary group will affect the change of these characteristics. For example, people are desirably in multivalence and present on the framework of thing and to introduce one or more PEGs (PEG) group and will improve hydrophily and water-soluble, increase molecular weight and molecular size, and, depend on the not person's character of (unPEGylated) framework of PEGization, can increase retention time in the body. In addition, people expect that PEG can reduce antigenicity, and improve the whole rigidity that thing is presented in polymerization by the hydrogen that is incorporated on the solvent molecule (such as water). Overlapping region similar between the characteristic of framework and the auxiliary group that can affect these characteristics is apparent to those skilled in the art.

Can improve multivalence presents the water-soluble and/or hydrophilic auxiliary group of thing and is actually used in the present invention. Therefore, improve multivalence with auxiliary group and present the water-soluble and/or hydrophily of thing and belong to the scope of the invention, these auxiliary groups comprise, for example, PEG, alcohol, polyalcohol (such as glycerine, propoxyl group glycerine, carbohydrate, comprise monose, compound sugar and polysaccharide, etc.), carboxylate, poly-carboxylate (such as polyglutamic acid, polyacrylic acid etc.), amine, polyamine (such as polyglycine, poly-(aziridine) etc.). Being used for improving water-soluble/hydrophilic auxiliary group in preferred embodiment is polyalcohol. Auxiliary group is PEG in particularly preferred example.

Present in multivalence and to mix lipophilicity and/or the hydrophobicity that the lipophilicity auxiliary group is presented thing with raising in the thing structure, this point belongs to the scope of the invention. The lipophilic group that is used for the present invention's practice includes, but not limited to aromatic group and many ring aromatic groups. Term used herein " aromatic group " refers to aromatic hydrocarbon and heteroaromatic hydrocarbon. Aromatic group can be not by or replaced by other group, replace but allowed their covalency to adhere to the group that multivalence presents on the thing by one at least. Other group that is used for the present invention's practice does not comprise the derivative of fatty acid that does not form bilayer at aqueous medium.

The lipophilicity auxiliary group will be annular group in preferred embodiment, such as cyclic hydrocarbon radical or heterocyclic radical. Annular group will be 6 yuan of rings in other preferred embodiment, or two or more 6 yuan of rings that condense. Auxiliary group will be phenyl or naphthyl in particularly preferred example.

What belong to equally the scope of the invention is the purposes of auxiliary group, and these auxiliary groups are so that multivalence is presented thing can be impregnated in the capsule, such as liposome or micelle. Term " lipid " refers to that all can form double-deck derivative of fatty acid, it so that the hydrophilic segment of lipid material in water hydrophobic part towards bilayer. Hydrophily comes from phosphate radical, carboxylate radical, sulfate radical, amino, sulfydryl, the existence of the groups such as nitro. Comprised, the thing of presenting such as, but not limited to, long-chain after the saturated or unsaturated aliphatic hydrocarbyl moiety group has just obtained hydrophobicity, and these groups can be by one or more fragrance, and ring grease or heterocyclic group replace. Preferred lipid is phosphoglyceride and (nerve) sphingolipid, representative example comprises phosphatid ylcholine, phosphatidyl-ethanolamine, phosphatidyl silk ammonia, the acid phosphatidylinositols, phosphatidic acid, POPC, lysophosphatidyl choline, hemolytic phosphatidyl-monoethanolamine, DPPC, DOPC, distearyl-phosphatid ylcholine or dilinoleoylphosphatidylcholine. Other also belongs to the lipid group without phosphorus compound such as sphingolipid and glycosphingolipids compound. In addition, above-mentioned both sexes lipid also can with other lipid, comprise triglycerides and sterols, use after mixing.

Multivalence is presented the flexibility of thing and can be controlled by introducing huge and/or firm auxiliary group. Existence huge or firm group can stop rotating freely of key in the framework or the key between framework and the auxiliary group or the key between framework and the functional group. Firm group can comprise, for example, and the group of the ring that the mutability of its conformation is existed and/or the restriction of multikey. Other can bring the group of rigidity to comprise polymer-based group, such as oligomeric-or polyproline chain.

Rigidity also can obtain by static. Therefore, no matter auxiliary group is negative electrical charge or positive charge, and the auxiliary group of identical charges can force to be presented the thing framework and form and to make the each other maximum configuration of distance of identical charges. Make with the group of identical charges each other more approaching energy consumption framework will be remained on keep the configuration that separates with between the electric charge auxiliary group. In addition, will attracted on the group with its oppositely charged with the auxiliary group of opposite charges, and will enter intermolecular and molecule in ionic bond. This non-covalent bonding mechanism is put up holding frame the conformation that combines for the group that allows the tool opposite charges. After being added to auxiliary group on the framework, by deprotection reaction well known by persons skilled in the art, change pH; oxidation reaction, reduction reaction or other mechanism make auxiliary group with electric charge; perhaps, the process with unshielded potential electric charge belongs to the scope of the invention.

Huge group comprises, for example, thick atom or ion (such as iodine, sulphur, metal ion etc.) contain the group of thick atom, and many cyclic groups comprise aromatic group, non-aromatic group and contain the structure (being alkene and alkynes) of one or more carbon-to-carbon multikeys. Huge group also comprises side chain or straight chain type oligomer and polymer. The structural rigidity that the every increase unit formula of expectation branched chain type amount increases is more than straight chain type.

Rigidity is because the existence of annular group (such as cyclic hydrocarbon group, heterocyclic group etc.) produces in preferred embodiment. Ring is 6 yuan of rings in other preferred embodiment. Ring is aromatic group in preferred embodiment further, such as phenyl or naphthyl.

Present the antigenicity of thing and also belong to the scope of the invention by selecting carefully auxiliary group to change multivalence. May need in some applications to reduce the antigenicity that multivalence is presented thing. As mentioned above, sheltering group, such as PEG, is known in the artly can reduce unit price and the antigenic group of multivalent molecule. (people) may need to improve the antigenicity that multivalence is presented thing in other is used, and therefore, have just excited immune response. Auxiliary group can comprise and known in the artly can improve the immunogenic group of haptens in these are used. Being suitable for improving multivalence presents the immunogenic group of thing and includes, but not limited to key hole chirp keyhole limpet hemocyanin (KLH) and the such protein of bovine serum albumin(BSA) (BSA). Other can improve multivalence, and to present the antigenic group of thing will be well known by persons skilled in the art.

Distributing in Half-life in vivo and the body is many character of molecule, comprises molecular size, molecular weight, electric charge, hydrophobicity/hydrophily, antigenicity, biological degradability and multivalence are presented the existence of target group on the thing or are not existed, function. Here used term " target group " refers to cell receptor is had the group of affinity. Thereby these character make the half-life of administered compound in the body or the method can obtain desired change that distributes is that materia medica and medical chemistry field are known by changing. Identify the method for new group A

The present invention also provides synthetic method (the Horton. D. as the oligosaccharides group of group A, " carbohydrate chemistry and biochemical development (Advances in Carbohydrate Chemistry and Biochemistry) ", Academic Press:San Diego, 1995), can synthetic protein analog (Maassen, J.A.; Terhorst, C., " european journal of biological chemistry " (Eur.J.Biochem.), 1981,115,153-8; Wang, the people such as K.S., " Journal of Virology " (J.Virol.), 1992,66,4992-5001; Mastromarino, the people such as P., " hereditary Journal of Virology " (J.Gen.Virol.), 1987,68,2359-69). The computer based design tool also can be used to synthetic group A (Jorgensen, W.L., " chemical field: organic chemistry " (Chemtracts:Org.Chem.), 1995,8,374-6).

The present invention also provides the method at the new group A of the conceptive discovery of multivalence effect. For example, single bacteriophage in the phage display library, single pearl string in single safety pin (pins) in the space addressing combination array (spatially addressed combinatorial array) or the combinatorial libraries that produced by " mix and division " method, each is only with a kind of group, and this group is presented in the multivalence mode. Therefore, these storehouses are directly screened in the combining form of the B that can present multivalence, and combining form has, and such as simple adhesion, or cause the adhesion of special result, as infecting cell death, hyperplasia, the generation that form changes or easily detects acceptor such as green fluorescent protein. A typical resin bead can be born about 100pmol or the individual copy of about 60,000,000 (trillion). Be not all these copies all on the surface of all solids carrier, but carrier can be chosen as, and is for example, porous or surface-functionalized. Comparatively speaking, bacteriophage can show 3-1000 part peptide group copy from the teeth outwards, and this depends on the coat protein that the storehouse condenses. Being decided to be is worth any group A that further analyzes next will carry out dissolubility, the test of polymerized form aspect.

As an example, concrete adhesion has bacterium, the pearl string of fungi or neutrocyte to carry the group that can present as multivalence of the present invention the thing group in the storehouse. There is approximately the storehouse of 1,000,000 different peptides can adopt the division synthetic method to make, and can obtains the panoramic storehouse of other type compound. The structure of selected group will be determined by building-up process, perhaps available ordering, and mass spectrum is separated curling (deconvolution) or coding method and is obtained.

For example, (for example can use the peptide storehouse of demonstration bacteriophage, referring to Doyle, M.V. wait the people, " utilize blood platelet and whole cell to select peptide part (The Utilization of Platelets and Whole Cells for the Selection of peptide Ligands from Phage Display Libraries) from phage display library ", " combinatorial libraries: synthetic, screening and application potential " (Combinatorial Libraries:Synthesis, Screening, and Application Potential), Cortese, R., Ed.; Walter de Gruyter:Berlin; 1996; Pp.159-174; The people such as Fong, " drug development " (Drug Dev.Res.), 1994,33,64-70; The people such as Goodson, " Proc. Natl. Acad. Sci.USA " (Proc.Natl.Acad. Sci.), 1994,91,7129-33). The bacteriophage (every 3-5 copy) that can select in an example (" elutriation ") PIII modification interacts with closing section's Activated platelet, thereby identifies 5 levels and else combine hematoblastic peptide group. Test with regard to the interactional inhibition problem of bacteriophage-blood platelet from the synthetic peptide that the bacteriophage sequence is derived, in order that prevent platelet aggregation, although the IC that observes₅₀Be worth very low.

Had document discussed with efficient glycosyl donor carry out the combination of polysaccharide synthetic (people such as Liang, " science " (Science), 1996,274:1520). This technology depends on different sulfoxide as glycosyl donor, and it has very high reactivity, and it is synthetic to allow to carry out polysaccharide with very high productive rate, and in a word, this is a fundamental characteristics of combinatorial libraries. Can the pearl string be that the basis creates (combination) storehouse, and in order in solution, to interact and will to screen the storehouse with the multivalence agglutinin. Adhesion determination method or agglutination assay can be used for this system.

Rational drug design technology, comprise computer based design tool (Jorgensen, W.L., " estimate the new method (A new method for predicting binding qffinity in computer-aided drug design) of binding affinity in the Computer-Aided Drug Design ", " chemical field: organic chemistry " (Chemtracts:Org.Chem.), 1995,8,374-6) can be used for differentiating the group A of multivalence in presenting. The adhesion of group A

Group A can be adhered on the framework with the mode that permission group multivalence is presented or form the part of framework. Adhesion can be directly, for example, and not by attachment, or indirectly, namely by attachment adhesion group A. Group A can adhered on the monomeric unit before or after the polymerization. The present invention is unqualified to the order of adhesion step, as long as the formed thing of presenting shows group A in the multivalence mode. For example, group A can at first adhere with attachment, and then, attachment-group A part is adhered with framework again, and vice versa. Perhaps, attachment is adhered with framework first, and then, group A partly adheres with framework-attachment again. Also can group A be mixed multivalence with the difunctionalization monomer with group A and present thing, then with group A polymerization. Perhaps, can prepare and have reactive polymer, namely prepare one and be activated to carry out the polymer of coupling reaction, then make group A and this polymer reaction (" pre-priming reaction " technology).

Preferred example is that attachment is adhered on the group A. As mentioned above, attachment comprises the group part that group A can be located or is adhered on the framework. Attachment is the part that is independent of group A and framework in preferred example. The example of preferred attachment type is primary amine can be stayed attachment not connect terminal attachment. Then, the group of modification can be adhered on the activating carboxy acid of polymer, forms acid amides and connects. Amido link is relatively stable, extensively occurs at nature, and can make very clearly with the method for having developed in a large number. In addition, can obtain with special attachment extra advantage.

Cationic polymer has more antigenicity than anionic polymer. Cation-modified part can use to guarantee to consume all modified groups with limited quantity, only stays excessive hydroxy-acid group at polymer, and like this, polymer has a positive effect to water-soluble. As mentioned above, attachment also makes certain space interval between group A and the framework. Attachment is returned multivalence and is presented thing and give flexibility in some instances, for example, and the flexible movement of group. The typical attachment of presenting thing for the present invention comprises poly-(glycine), alkylamine such as ethamine, hydrocarbon part, thioether, poly-(ethyleneimine) and poly-(aspartic acid), and polyethers such as PEG chain.

" pre-priming reaction " method allows with the not isoplastic monomeric unit tectonic framework of presenting that limits the mole component, such as polymer. For example, the method can be used the Acibenzolar of poly-(acrylic acid), such as poly-(N-acryloyl group succinimide), the i.e. pNAS that is polymerized by N-acryloyl group succinimide. Next, the DMF (solution) of the NHS ester of pNAS can from different primary amine (such as R₁NH ₂And R₂NH ₂) reaction, form amido link. By using excessive NH₃Or OH^-Processing can change into respectively acid amides or carboxylic acid with being left unreacted ester group. It is believed that adopting these groups after this method will be at random with respect to the distribution of polymer backbone. And owing to detected the group of ipsilateral not to the impact of usefulness, extent of polymerization (sum of monomeric unit in polymer) can keep constant. With¹H-NMR spectrometer and combustion analysis can be estimated the efficient that forms the reaction of acid amides between amine and the pNAS. The completion rate of this reaction is decidedly superior to 90%.

Directly another example of " copolyreaction " mixture of can be used on the acrylamide monomer derivative among the backflow THF is explained (people such as Spaltenstein, above-mentioned quoted passage). Copolyreaction can be introduced two main unrestricted variables in polymerisation: (i) between the acrylamide that replaces of different N the unknown difference of the proportionality constant of copolyreaction may to cause group (A) be not unique distribution with respect to the skeleton of polymer; (ii) polydispersity of polymer, succession and length may change because of the change of side chain. Owing to relate to polymerization process (breeding; Termination procedure) ratio is variant, therefore above-mentioned phenomenon can occur. If the pA (R) of unique architectural feature need to be arranged, then " pre-priming reaction " method may be preferred.

The method activation that polymer is had high yield and is easy to during the course remove accessory substance in preferred embodiment. What other need to be considered comprises accessory substance and the reactivity of the group of adhering, and causes the trend (as with gathering (amino acid)) of racemization.

For example, there are a series of chemical methodes to activate the carboxylic acid that reacts with amine. The relative ability of activating carboxy acid's reaction successively decreases with following order; RCOHal＞(RCO)₂O+RCON ₃＞ RCO ₂R＞RCONH ₂＞RCOR, and increase along with the nucleophilie nucleus ability of the amine that reacts. What use in preferred embodiment is inner-acid anhydride.

Since on polymer backbone with acid anhydrides or succinimide, so when reacting with amine, just be easy to form amido link. Intramolecular acid anhydride in the peptide can use alpha-amido-N-carboxylic acid anhydrides or thiocarboxylic acid acid anhydride to form.

Also can use mixed acid anhydride in some instances, for example, carbodiimides etc. The method that is generally used for the formation mixed acid anhydride of chemistry of peptides comprises formation carbodiimides (for example, with dicyclohexyl carbodiimide (DCC) or 1-ethyl-3-(3-dimethylamino) propyl carbodiimide diimine (EDC)). EDC can be used for water-soluble polymer. Other acid anhydrides that exemplifies can be made by reagent such as chloro-formate or quinoline (EEDQ, IIDQ).

As mentioned above, the carboxylic acid azide also can use in other example. Need to before reaction, make azide in the situation of using the carboxylic acid azide. In other example, can use imidazoles, such as carbonyl-imidazoles.

In the other example, can use and resemble the compound that p-nitrophenol or N-hydroxy-succinamide can form Acibenzolar like this.

Used polymer contains hydroxyl in the other example, therefore can use the method for derived hydroxy groups. This method is well known by persons skilled in the art.

Perhaps, make multivalence with combined method and present thing. In brief, the solid-state combined method of available standard generates with group A₁To A_nThe skeleton storehouse. Combinatorial libraries is presented the thing array combination by synthetic multivalence and is formed, and wherein the multivalence in the array is presented the composition of thing, and structure, character, function etc. are all different each other. When the manufacturing multivalence is presented the thing array, people especially can change the chemical constitution of frame part except other content, the chemical constitution of auxiliary group, the chemical constitution of interval group, the chemical property of frame part, the chemical property of functional group part, the chemical property of auxiliary group, the chemical property of interval group, the amount of the frame part of presenting, the amount of the functional group part of presenting, the amount of the auxiliary group part of presenting, the amount of the interval group of presenting, number and/or the amount of the different frames part of presenting, number and/or amount that the difference in functionality regiment headquarters of presenting divides, the number of the different auxiliary groups of presenting and/or amount, number and/or the amount of the different interval group that present, the character of connecting key and quantity are (for example between the various compositions, the character of the connecting key of interval group), response parameter (such as reaction dissolvent, reaction temperature, reaction time, reaction initiator, catalysts reacts residing gaseous environment, reaction pressure, the speed of cancellation reaction etc.), the stoichiometry of various compositions; Present the order that heterogeneity adds, etc.

Therefore, the invention provides in an instantiation and prepare the method that multivalence is presented the thing array, the method comprises: (a) the first multivalence is presented the frame part of first activation of thing and the frame part that the second multivalence is presented first activation of thing and be added in the first and second reaction vessels; And (b) the first multivalence is presented first functional group part of thing and first functional group that the second multivalence is presented thing partly is added in the first and second reaction vessels, form at least two kinds of different multivalence and present thing. Allow to want preactivated reactant to react. This process optional with other composition (such as frame part, functional group part, auxiliary group, interval group etc.) and/or different response parameters (such as different reaction temperatures, catalysts, reaction dissolvent etc.) repeat, present the thing array to form huge multivalence.

Multivalence is presented the thing array in case form, and just can present the due useful quality of thing according to multivalence and screen and/or measure. The character that can screen includes, but are not limited to: biologically active, in conjunction with affinity, biological property, pharmacological properties, oral administration biaavailability, circulating half-life, agonist activity, antagonistic activity, dissolubility etc. The useful quality of next or simultaneously multivalence being presented the thing array is screened. In addition, can screen array then and there, perhaps, need not work as field type and multivalence is presented thing screen, for example, from substrate, take out and screen again after multivalence is presented thing. In case differentiate and know that just can prepare on a large scale the multivalence with useful quality has presented thing. The interval group

In another instantiation of the present invention, the interval group, or rephrase the statement, the attachment group is between framework and functional group and/or framework and auxiliary group.

The construction of this instantiation of the present invention has following general formula:

R ¹{-R ²(-R ³) _n} _m (II)

R ¹{-R ²(-R ³) _n} _m{-R ⁴(-R ⁵) _s} _t (III)

Above symbol R in two structural formulas¹Being represented as the interval group provides the multifunctional frame in a plurality of adhesions site. Comprise dendrimer, polypeptide, polysaccharide etc. generally are used as this framework at interior polymer. Because the existence in a plurality of adhesions site, skeleton plays amplification for functional group and/or auxiliary group.

Symbol R in two general formulas in the above²And R³Represent respectively interval group and functional group. Symbol m representative is bonded at the quantity of the functional group on each interval group. Symbol n represents interval group and the quantity that adheres to functional group framework on relevant with them. As mentioned above, usually to present 10 times of sum of functional group on the thing and/or auxiliary group than multivalence also many for the quantity of the quantity of the quantity of functional group and auxiliary group and interval group.

Symbol R in general formula III⁴And R⁵Represent respectively interval group and auxiliary group. Symbol s representative is bonded at the quantity of the auxiliary group on each interval group. Symbol t represents interval group and the quantity that adheres to auxiliary group framework on relevant with them. It is also many that the quantity of the quantity of functional group and interval group is presented 10 times of sum of functional group on the thing than multivalence usually. For example, " m " can be 6 and n can be " 3 ", illustrates that always having 18 functional groups exists.

In the example of general formula I I representative, only have interval group/functional group construction to adhere on the framework. Interval group/functional group and two kinds of constructions of interval group/auxiliary group all adhere on the framework in the example of structural formula II I representative. Interval radicals R in general formula III²And R⁴Both can be the same or different, and existed with 1: 1 mol ratio or different mol ratios approximately. For the purpose of brief for the sake of simplicity, only mention symbol R in the following discussion², still, should be appreciated that R²And R⁴All comprise under discussion. Correspondingly, relate to R³The discussion of (functional group) also comprises R⁵(auxiliary group).

The interval radicals R²Can be any in a variety of molecular structures, and be bifunctional at least, to allow to connect simultaneously R¹And R³, optional process linking group connects. The interval group be stable or can be in biotic environment cracking in the body. (be R in end position³End) there are a plurality of interval groups in conjunction with functional group can hold a plurality of functional groups and/or auxiliary group. This interval group plays a part amplification in the mode of similar framework, although just on less degree. Other is used for interval of the present invention group can only at one end have single functional group, and n will be 1 in this case, and m will be greater than 10. The cleavable, especially attractive character is hydrophily in above-mentioned body. Other useful character is the ability that reduces antigenicity and increase molecular weight. Interval group with other character also can use to strengthen its advantage, because this all is apparent for those skilled in the art.

R ³The functional group of representative is above-mentioned group A. The interval radicals R²It can be the straight or branched structure. Preferred R²Group is the group that straight chain is arranged in the structure, and this linear chain structure both can be whole interval group, also can be the skeleton of branched group. Straight chain can be the chain that carbon atom consists of, and also can be that the centre is inserted with one or more hetero atoms, such as oxygen atom, and the carbochain of sulphur atom or nitrogen-atoms. This chain also can be replaced by aromatic group. The key that consists of chain can be singly-bound, two keys or triple bond, but preferred singly-bound. The length of chain is unrestricted, can have a variety ofly, and this depends on needed correlation between the quantity of functional group on the molecular weight of construction and the construction and/or auxiliary group. Optimum is the chain acquisition of 4-1000 atom by length usually, and preferred chain length is 6-100 atom, and most preferred is 10-50 atom. Said chain is the skeleton of interval group itself, and does not comprise atom, group or the side chain that is connected on the series winding atom that forms skeleton. But described chain is included in the end of the chain chain is connected to R¹And R³If on linking group---have linking group.

The interval group will have water-wet behavior in some example of the present invention, and gives construction this characteristic. Therefore, the interval group can be hydrophilic radical well known by persons skilled in the art. The example of hydrophilic radical has polyglycols, and optional quilt can increase or not increase hydrophilic group and replace. Preferred polyethylene glycol in polyglycols. Optional substituting group comprises, for example, and alkyl, alkoxyl and hydroxyl. Unsubstituted polyethylene glycol is particularly preferred. The length of the optional polyethylene glycol that replaces is unrestricted, can be a variety of. The selection of length mainly is based on these considerations of hydrophily that will make construction reach required molecular weight and will give required degree to construction. Adopt molecular weight to be about 100-20 in great majority are used, 000 daltonianly polyolefin-basedly will obtain optimum efficiency, 200-1 more preferably from about, and 000 is daltonian polyolefin-based.

The present invention by the interval group to construction contribution body in the example of cleavable the interval group can contain any group that can be used as the part of its chain, this chain can be in biological fluid with the faster speed cracking of structure of more this than not having (interval) group. The speed of accelerating cracking can improve from multivalence presents the functional group of thing and/or the speed of auxiliary group split-frame. Such separation can or improve the functional group activity for reducing toxicity. Although the present invention is the degree that improves of rate of cleavage without limits, can be in biological fluid in administration 24 hours at least the interval group of the about 10% cleavable group of cracking be preferably, the interval group of cracking about 35% at least most preferably. Preferred cleavable group is that the ester connection is connected with disulphide.

The interval group was both given hydrophily to construction in another example of the present invention, comprised again above-mentioned cleavable group.

The structural formula of interval group has a variety of. Below discussion only for the example of a kind of interval group of being provided for the present invention practice, rather than to limit kind for the interval group of the present invention's practice. Structural formula of giving hydrophilic interval group to construction is that m is 1 structural formula among above-mentioned general formula I I and the III, the R among general formula I I and the III²By following structural IV, V or VI represent:

X-R ⁶-Y-R ⁷-Z (IV)

X-R ⁷-Y-R ⁶-Z (V)

X-R ⁶-Z (VI)

In the above in each structural formula hydrophilic segment by R⁶Representative, it is that molecular weight is about 100-20,000 dalton, 200-1 more preferably from about, 000 daltonian polyethylene group.

Radicals R in structural formula IV and V⁷Expression can increase the cleavable group of construction rate of cleavage in blood. This group is disulfide group S-S, or the ester group that is positioned in all directions, i.e. C (O)-O or O-C (O). In case R wherein²The construction cracking that is represented by structural formula IV, then the polyethylene glycol gene will with the framework R of polymerization¹Remain together. On the contrary, in case R wherein²By the construction cracking that structural formula V represents, then polyethylene group will remain with functional group and/or auxiliary group.

Symbol X, the dying links group that Y and Z representative link together the R group. The character of these linking groups is unrestricted, and if their selection adopt the construction synthetic method to determine it will will be very easily. Term 'inertia' used herein refers to basically that in clinical or diagnostic procedure using this construction is nontoxic basically in the required general time period, without immunogenic and be stable with respect to cracking or division. The example that is used for the dying links group of this purpose has alkyl amino or aminoalkyl, such as (CH₂) _q-NH and NH-(CH₂) _q, carbamoyl, such as NH-C (O)-O and O-C (O)-NH, and alkyl-carbamoyl or carbonylic alkyl, such as (CH₂) _q-NH-C (O)-O and O-C (O)-NH-(CH₂) _q Symbol q in these groups can change, but in most cases is generally 1-10, preferred 2-4, particularly preferably 2. These groups can so define in specification of the present invention, and namely the end position atom of X or Z can be respectively R¹Or R³Intrinsic. For example, the end position NH group in X or the Z definition can be by R¹Or R³On amino functional group or other band N group that can react the NH that forms linking group consist of. Equally, the end position is the O atom.

The structure of structural formula IV definition can exemplify as follows:

The structure of structural formula V definition can exemplify as follows:

Symbol " PEG " refers to therefore, vinyl is positioned at two ends with the polyethylene glycol segment of separative end position hydroxyl in above-listed 6 structural formulas. The interval group that does not contain the similar common primitive formation of PEG also is used to the present invention's practice.

As the distortion of structural formula IV, support the structural formula of the interval group of 11 or 11 above functional groups to be represented by structural formula VII.

X-R ⁶-Y’(-R ⁷-Z) _r(VII) R in this structural formula⁶，R ⁷With X as defined above, Z refers to (CH₂) _q-NH. Symbol Y ' represents the following formula group,

O-C(O)-NH-(CH ₂) _q-NH ₃(VIII) wherein, q is 1-3, is connected in (the CH of structural formula₂) _q-NH ₃Two or more H atoms on the C atom in the part are substituted basic NH-(CH₂) _s-NH substitutes, and wherein s is 2-4, and substituent quantity is determined with above-mentioned method about m and n among structural formula II and the III. As a result, the interval group is the branched structure that contains the reactive NH group of two or more be used for adhering functional group and/or auxiliary groups.

Symbol r among the structural formula VII is 0 or equals n. When r is 0, do not have the cleavable group in the group of interval, and when r was not 0, each the functional group NH on Z attachment (functional group or auxiliary group adhere on the Z attachment) included the cleavable group.

Q is 2-6 in the preferred embodiment of structural formula VIII, and most preferred q is 2 or 3.

As further variation, the end position CH of structural formula VIII₃Can be replaced by OH or SH. The result causes being conducive to reactive OH or the SH group of the adhesion of functional group and/or auxiliary group.

R among structural formula II and the III²Other structural formula defined by structural formula IX:

X’-R ⁸-Z‘ (IX)

R in structural formula IX⁸The following formula group,

(CH ₂) _o-R ⁹-(CH ₂) _p(X) wherein, R⁹The R with said structure formula IV and V⁷The cleavable group of identical definition, i.e. disulfide group S-S, or the ester group that is positioned in all directions, i.e. C (O)-O or O-C (O). Subscript o and p are identical or different, equal 0 or positive integer, but o and p sum are at least 2.

Symbol X ' and Z ' among the structural formula IX are identical or different, are the inertia groups of the dying links group in the similar said structure formula. Preferred X ' and Z ' are NH-C (O), C (O)-NH, NH-C (S) and C (S)-NH.

Functional group in these structural formulas on the single construction of structural formula II or III and/or the quantity of auxiliary group and arrangement can alter a great deal. The quantity of functional group equals the product of m and n. The quantity of auxiliary group equals the product of s and t. In a word, all constructions are that above-mentioned one or two product is all greater than 10 construction.

Construction of the present invention can be synthetic according to conventional coupled reaction well known to those skilled in the art. In example, wherein by the framework R of polyamines radical functino¹Be provided as follows. Skeleton refers to (AMP) (NH in this example₂) _x, " AMP " refers to allow the amplification effect of a plurality of intervals group and functional group and/or auxiliary group adhesion, and the x representative is corresponding to the numeral of n among structural formula II and the III.

The adhesion of polyethylene glycol (PEG) interval group and amplifier (amplifier) can be used the Acibenzolar of PEG, such as the α of PEG, and ω-biconjugate nitro-phenoxy ester realization,Wherein, " PEG " is that polyethylene glycol deducts end position hydroxyl, with top definition. Excessive PEG ester derivant X and (AMP) (NH₂) _xReaction generates following intermediate:

Then, this intermediate and excessive N₂H-(part) reaction, this part refers to contain after functional group is derived the part of reactive amino. Product has following moulding:

In obtaining other flow process of similar product, by with diamines such as NH₂-(CH ₂) ₂-NH ₂Reaction, above-mentioned intermediate X I is converted to the second intermediate with end position amido. The second intermediate has the following formula structure:

Then, the part of intermediate X IV and activated carboxylic, such as the acid anhydrides of part, reaction generates the group that the following formula structure is arranged,

Cleavable group such as disulfide group can be by intermediate X and the inner diamines that contains disulfide group, such as cystamine disulphide NH₂-(CH ₂) ₂-S-S-(CH ₂) ₂-NH ₂, reaction generates another intermediate and introduces.Then can with the ligand reaction of carboxylic acid activation, generate

Also have many other methods. Contain the construction that does not have the cleavable of PEG ester in the group of interval in order to make, for example, the amplification polymer that can derive and contain amine or hydroxyl is made hydroxy-acid group as functional group. By polymer and maleic anhydride, succinyl oxide or glutamic acid anhydride reactant just can be realized above-mentioned preparation at an easy rate with the reaction process of having set up. To can pass through with the part of deriving of the combination of polymers of deriving part and amino alcohol HO (CH with the isosulfocyanate radical group₂) _nNH ₂Reaction, the end position hydroxyl on the displaced ligands forms. Then by conventional method reagent, such as dicyclohexyl carbodiimide or carbonyl dimidazoles, can activate the carboxylic acid on the polymer of deriving, and be connected to obtain ester with the ligand reaction of deriving. Have polymer and the structure between the part of amplification partly to play interval group effect, the length of this interval group is by being used for CH in the amino alcohol of derived ligand₂The quantity of group determines.

In contrary other method of ester of preparation, part amino carboxylic acid HO₂C(CH ₂) ₂NH ₂Derive, rather than derive with amino alcohol. Then with the part of dicyclohexyl carbodiimide or carbonyl dimidazoles activation gained carboxylic acid derivatives, then part directly is coupled to hydroxyl and amplifies polymer.

In above-mentioned two kinds of methods, if necessary, can originally belong to the impact that the amido that amplifies polymer or hydroxylic moiety protect to avoid coupling reaction with selected. By conventional method, carry out famous acetylization reaction with acetic anhydride, just can realize at an easy rate protection.

Being useful on the part that adheres to the functional group on the group of interval can prepare with conventional method. The well-known part that exemplifies can be made at an easy rate with method known to those skilled in the art. Also keep its part whole or most of binding affinity after preferentially being chosen in derivatization reaction. Response variable

As mentioned above, prepare multivalence of the present invention and present the formation that a kind of method of thing relates to combinatorial libraries, this storehouse is presented the thing array by synthetic multivalence and is formed, wherein, the multivalence in the array is presented thing and formed, and structure, character, the aspects such as function are different from other array. When the manufacturing multivalence was presented the thing array, people especially can change the chemical constitution of frame part, the chemical constitution of auxiliary group, the chemical constitution of interval group except other content; The chemical property of frame part, the chemical property of functional group part, the chemical property of auxiliary group, the chemical property of interval group; The amount of the frame part of presenting, the amount of the functional group part of presenting, the amount of the auxiliary group part of presenting, the amount of the interval group of presenting, number and/or amount that the difference in functionality regiment headquarters of presenting divides, the number of the different auxiliary groups that present and/or amount, number and/or the amount of the different interval group that present, the character of connecting key and quantity between the various compositions (for example, the character of the connecting key of interval group); Response parameter (such as reaction dissolvent, reaction temperature, the reaction time, reaction initiator, catalysts reacts residing atmospheric pressure, reaction pressure, the speed of cancellation reaction etc.); The stoichiometry of various compositions; Present the order of heterogeneity, etc. Each reacted constituent (for example, frame part, functional group part, auxiliary group, interval group etc.) and used the whole bag of tricks will be discussed below in more detail.

The polymer that activates in a method and functional group react basically to consume on the aggregation framework mode of all activated groups. The stoichiometric relationship of functional group and activated group is at least 1: 1 in this instantiation. If 1: 1 ratio is not enough to basically consume whole activated groups, it is excessive then functional group can be added to.

The addition of functional group also can be not enough to consume whole activated groups in another method. After reacting with the first functional group, add the second functional group or auxiliary group, and then continue reaction. Be included in also having within this example: adding more than one functional group, both can be mixed form, also can sequentially add, and then add auxiliary group, perhaps, adds more than one auxiliary group, to mix or sequential system.

In another method, basically consume activated group by the functional group that adds capacity. After the first functional group is adhered on the framework, with the frame part reactivation, add the second functional group or auxiliary group. Each step functional group of reaction process can be a kind of functional group in this example, the mixture of difference in functionality group or the mixture of functional group and auxiliary group, and wherein, auxiliary group also can be single auxiliary group or the mixture of auxiliary group.

Activated group consumes by the mixture of two kinds of functional groups of interpolation or the mixture of at least a functional group and at least a auxiliary group basically at least in another approach.

Activated group and at least a functional group of the overwhelming majority (than basically all lacking) in another method, the mixture reaction of at least a auxiliary group or at least a functional group and at least a auxiliary group. Activated group remaining after first step reaction is basically by adding another kind of functional group, and the mixture of auxiliary group or functional group and auxiliary group consumes.

Functional group in some instances, the mixture of auxiliary group and/or functional group and auxiliary group is added in the framework solution of activation with dry powder or neat liquid form. Functional group in other example, the solution of the mixture of auxiliary group and/or functional group and auxiliary group is added in the polymer solution of activation. Functional group in other examples, the solution of the mixture of auxiliary group and/or functional group and auxiliary group is added in the polymer, and wherein polymer exists with neat liquid or dry powder form.

Be used for solvent of the present invention comprise with the activated group that participates in reaction (that is, basically not and the framework of activation with very large quantitative response) and the polymer of activation and functional group or auxiliary group between compatible all solvents of the character of reacting. These solvents comprise, for example, and water, methyl-sulfoxide (DMSO), dimethyl formamide (DMF), alcohol, ether, ketone, hydrocarbon, aromatic hydrocarbon, and the mixture of any ratio of these solvents.

Control (reaction) temperature can reach the purpose of reaction speed between control activation framework and functional group and/or the auxiliary group in other examples. Other response parameter in other example (such as reaction dissolvent, reaction temperature, the reaction time, reaction initiator, catalysts reacts residing atmospheric pressure, reaction pressure, the speed of cancellation reaction etc.) can change and optimization.

" handle (handle) " in the other example on functional group or the auxiliary group activated by above-mentioned activated group, then with framework on activated group or framework on not activated partial reaction. Here used term " handle " refers to reactive group, such as carboxyl (acid or salt), hydroxyl, sulfydryl, acid amides, carbaminate, amine, ketone, aldehyde, alkene, alkadienes, aromatic rings etc. The appropriate combination of the handle of framework connected reactive moieties and functional group or auxiliary group it will be apparent to those skilled in the art that.

Those skilled in the art should understand that, although top discussion is relevant with the use of activated polymer and functional group, but all above-mentioned examples all are applicable to relate to and use monomer (such as the monomer of not deriving, the monomer of deriving with the interval group, monomer of deriving by the interval group directly or indirectly with auxiliary group etc.) and the functionalization monomer (as using functional group, i.e. R³, the monomer of deriving by the interval group directly or indirectly) method. Substrate and be delivery system

Here used term " substrate " refers to have the material of rigidity or semi-rigid surface, perhaps has dimple, hole, nest, the material of irrigation canals and ditches etc. Although need in some instances to use with, for example, dimple, hole, convex surface, the different materials of etched irrigation canals and ditches etc. consist of physically separated synthetic zone, at least one surface is level and smooth basically on the substrate in many examples. Substrate itself just has hole in some instances, convex surface, and etched irrigation canals and ditches etc. consist of all or part of of synthetic zone.

More specifically, substrate can be organic matter, inorganic matter, and biology, abiotic or their any combination, with particle, lines, sediment, gelatin, coverlet, pipe, ball, container, capillary, pad, sheet, film, plate, the shapes such as sheet glass occur. Substrate is preferably level and smooth, but various different surfaces configurations also can be arranged. For example, substrate can have convex surface or concave surface, and multivalence of all kinds is presented thing and can be synthesized on male and fomale(M﹠F). Substrate and surface thereof preferably can form rigid carrier, can carry out above-mentioned reaction in the above. Substrate can be material miscellaneous, for example, and polymer, plastics, Bohemian glass (pyrex), quartz, resin, silicon, silica or based on the material of silica, carbon, metal, unorganic glass, mineral crystal, film etc. Also be obvious for a person skilled in the art based on other substrate of viewpoint of the present disclosure. The surface of solid substrate can be made of the material identical with substrate, perhaps, is made of different materials, and namely substrate is by different material dressings. And, on substrate surface, can also coat adsorbent (such as cellulose), the part of being concerned about is just presented in the above. Only substrate and substrate surface material determine that by the grade of the material that will synthesize under any circumstance its selection all is apparent for a person skilled in the art. Substrate suitable in preferred embodiment comprises, for example, titer plate (such as 96 hole titer plates) or rack for test tube can be placed on the rack for test tube and hold multivalence and present that each presents the test tube of the sufficient amount of thing in the thing array.

Reactant composition at each conversion zone must often want it to prevent from moving on to adjacent conversion zone. The simplest method is by stay enough intervals between the conversion zone on the substrate so that each composition can not spread mutually between conversion zone ensures, suitable obstacle perhaps is set between the conversion zone on the substrate ensures. In a method, plant equipment or physical arrangement have been determined each conversion zone on the substrate. For example, wall or other physical obstacle can be used to prevent that the reactant composition from moving on to adjacent conversion zone from a conversion zone. This wall or physical obstacle can be removed after synthetic having carried out. It should be appreciated by those skilled in the art that and before the screening material array, remove wall or physical obstacle is favourable sometimes.

In other method, for example, hydrophobic material can be used to wrap up each conversion zone zone on every side. These materials can prevent that sealing (and other polar substances) flow of solution is to the substrate in the adjacent conversion zone. Certainly, if use non-aqueous or non-polar solven, the surface coatings that needs are different. In addition, by selecting suitable material (such as substrate material, hydrophobic coatings, reaction dissolvent etc.) people can control the contact angle of drop and substrate surface. Owing to solution makes reaction zone zone on every side wet in conversion zone, so need large contact angle.

Each reacted constituent that is a small amount of accurate measurement in the delivery system in the present invention is presented in each reaction zone. These can be finished with the various technology of presenting. For example, each reacted constituent can be presented to interested reaction zone with drop or powder type by distributor. For example, can adopt trace commonly used to inhale moving device and distribute various droplet sizes from capillary. Distributor also can be a kind of in the used type on the ink-jet printer commonly used. This ink-jet allocating system comprises, for example, and pulse die mould allocating system, bubbling ejection-type allocating system and slot injection type allocating system. These ink-jet allocating systems can be presented various droplet sizes. In addition, these allocating systems can be manual, also can be automatic or automanual, for example, adopt telecontrol engineering.

In other example, can reaction solution be delivered on the substrate from storage tank with the electrophoresis pump. Tiny capillary links to each other the storage tank of reactant with the nozzle of distributor in this instrument. Provide potential difference in two end electrodes capillaceous. Just as is known to the person skilled in the art, the transmission speed of all kinds of chemical substances such as charge density, is transmitted the size and dimension of material by various physical propertys in the electrophoretic medium of electric potential gradient is arranged, and the physics and chemistry character of transmission medium itself determines. Size capillaceous under suitable electric potential gradient condition, the rheology and the hydrodynamics that reach transmission medium flow and will set up in capillary. Therefore, containing to some extent, the large fluid streams of the reactant of care is pumped into substrate from storage tank. By regulating the relative position of substrate and electrophoresis pump nozzle, reaction solution can accurately be presented to predetermined reaction zone on the substrate.

The relative position in available various conventional system calibration distributor and appropriate reaction district. This type systematic is widely used in microelectronics instrument manufacturing and test field, and they can be sent to each reaction zone with the speed up to 5,000 of per seconds with the drop of reacted constituent. The translation of this type systematic (X-Y) precision is better than in the 1 μ m. Available the whole bag of tricks known in the art is calibrated the distributor of this type systematic and the relative position of substrate. For example, only utilize only one or two reference point of substrate surface, with " flight path supposition method " each reaction zone on the substrate is located. Reference identification in this type systematic can be used electric capacity, and resistance or optical pickocff are accurately determined. Perhaps, use " video " system that camera is housed.

Available for the similar system calibration distributor in magnetic and optical storage media field and the relative position between the interested reaction zone in another example. For example, reacted constituent is identified the reaction zone of deposition by its track and sector on dish-shaped substrate. Then, when dish-shaped substrate rotation, distributor is moved on on the suitable track. Locate the drop of afterreaction solution below distributor just can emit when suitable reaction zone.

Reaction zone can further be determined by the sand holes of substrate surface in some instances. When probe other sensor must contact or when slipping over substrate surface this point seem advantageous particularly. Sand holes can also make distributor point to interested reaction zone as identification marking.

Skilled person in the art will appreciate that multivalence presents the structure of thing, namely composition can be determined by synthetic process, perhaps by ordering, and mass spectrum, deconvolution, the methods such as coding obtain. This estimating techniques exist, for example, the people such as Thomson, " chemical abstracts (Chem. Rev.) ", 1996,96,555-600, in describe to some extent, this article is incorporated by reference in the lump at this. Pharmaceutical composition

The present invention relates to be for multivalence the pharmaceutical composition of delivery of therapeutic agents. This pharmaceutical composition contains above-mentioned multivalence presents thing and pharmaceutically suitable carrier. This multivalence is presented thing in instantiation of the present invention following structural formula:

(Y)-(X-A) _n

Wherein, Y is framework, and X is straight key or attachment, and A is functional group, and n is integer greater than 10, and the selection of this integer can make the group presented and the target binding site B of set have an effect. This is presented thing itself and can be used as the pharmaceutically suitable carrier of oneself. Multivalence is presented thing and can so be prepared in an example, for example, suitably select n, and make-(X-A) part is adhered to Y, and conform to the interface of containing target binding site B set by individual administration being made multivalence present thing, and cover target binding site B and gather. X connects gene in another example, and it is part independently, does not belong to the part of Y or A, and n is the integer greater than 10, and its selection makes presents thing to the multivalence after the individual administration and can conform to target binding site B set. Y is aggregation framework in another example, and the functional group that A is presented, X are linking groups, and n is integer greater than 10, and the selection of this integer is so that present thing to the multivalence after the individual administration and can be consistent with target binding site B set.

Term " pharmaceutically suitable carrier " is will comprise presenting the material that the function that it will be realized was used and can be realized to thing jointly with multivalence. The example of this class carrier comprises solution, solvent, decentralized medium, sustained release agent, emulsion etc. These purposes for the medium of pharmaceutically active substance service are well known to those skilled in the art. Any multivalence that is applicable to is presented other common carrier of thing and is all belonged to the scope of the invention. Should be appreciated that it is water miscible that many multivalence of the present invention are presented thing. Therefore, multivalence of the present invention is presented thing be mixed with pharmaceutical composition than easier with the material of other poorly water-soluble, problem still less.

The multivalence of " treatment effective dose " present thing refer to treatment can realize among the patient function that it will be realized must or sufficient amount. The treatment effective dose can change because of various factors, for example, and the target position point of general, the type of used composition (for example, be framework, or attachment or group A), patient's body weight, or the order of severity of symptom. Those skilled in the art can study above-mentioned factor, and the effective dose that multivalence is presented thing is made the decision that does not have unreasonable experience. " effective dose " that external or in vivoassay method also can favourable definite multivalence be presented thing. Those of ordinary skill all can be chosen an amount of multivalence and present thing for the said determination method.

The data of cultivating determination method and animal experiment research acquisition by cell can be used for determining the appropriate dose scope that is used for the treatment of the patient. The dosage of these medicaments preferably having within minimum toxicity or avirulent circulation or the tissue concentration scope, comprises ED₅₀ This dosage can change according to used formulation and method of administration within the specific limits. For all medicaments that are used for the inventive method, dosage can be cultivated the measurement result preresearch estimates according to cell, to obtain to comprise IC₅₀(reaching the concentration that suppresses the required test medicine of half maximum efficiency of symptom) is in interior concentration range. These data can be used to help to be identified for more accurately the dosage of human body. Available, for example, with high-efficient liquid phase color spectrometry blood concentration.

Dosage regimen also can affect the key element that consists of effective dose. Multivalence is presented alone administration of thing, also can with other agent administering drug combinations of living. In addition, dosage can be divided into several parts and interim dosage, every day or continue and take, or whole dosage continuous drip. And the dosage that multivalence is presented thing can increase or reduce in proportion according to the urgent degree for the treatment of situation.

The all right conventional method of pharmaceutical composition used according to the invention, formulated with one or more physiologically acceptable carrier or excipient as pharmaceutically suitable carrier. Therefore, multivalence is presented the medicament that thing and physiologically acceptable salt and solvate thereof can be mixed with in the following manner administration, for example, the local use, injection sucks or is blown into (by mouth or nose), or oral, cheek, stomach and intestine are outer or rectally.

Therefore, medicament of the present invention can be made and be suitable for selected concrete administering mode, such as whole body and topical, form. Preparation technique and prescription generally can be at Remmington ' s Pharmaceutical Sciences, Meade Publishing Co., and Easton finds among the PA. For the whole body administration, preferred injection system comprises intramuscular, intravenous, the interior and hypodermic injection of peritonaeum. For injection system, multivalence medicament of the present invention can be mixed with liquid solution, preferably prepares in the buffer solution of physical compatibility, such as Hank solution or Ringer solution. In addition, medicament can be mixed with solid form, again dissolves before the use again or is suspended in diluent, makes parenteral solution. Certainly also comprise lyophilized form.

For oral administration, multivalence is presented the pharmaceutical composition of thing and can be made with conventional method and pharmaceutically acceptable excipient, for example, and tablet or wafer form. Described excipient comprises adhesive (such as the cornstarch of in advance gel, PVP or hydroxypropyl methylcellulose); Filler (such as lactose, microcrystalline cellulose or calcium monohydrogen phosphate); Lubricant (such as dolomol, talcum powder or silica); Disintegrant (such as farina or sodium starch glycollate); Or wetting agent (such as NaLS). Tablet can be used the means known in the art dressing. The fluid composition of oral administration can be made, for example, solution, syrup or form of suspension also can be made first dry form, are mixed and made into liquid form with water or other suitable carrier again before the use. This fluid composition can be made with conventional method and pharmaceutically acceptable additive. Described additive comprises suspending agent (such as the D-sorbite syrup, the edible fat of cellulose derivative or hydrogenation); Emulsifying agent (such as lecithin or Acacia); Non-aqueous carrier (such as apricot kernel oil, grease, the vegetable oil of ethanol or fractionation); And anticorrisive agent (such as methyl p-hydroxybenzoate or propyl ester, sorbic acid). If necessary, said composition can also contain buffer salt, flavor enhancement, toner and sweetener. The composition of oral administration can be made and to control the appropriate dosage forms that active material discharges.

For inhalation, but composition used according to the invention usually can the administration from push-through packs or sprayer of aerosol spray mode, wherein can add suitable propellant, for example, dicholorodifluoromethane, Arcton 11, dichlorotetra-fluoroethane, carbon dioxide or other suitable gas. Dosage unit can measure by add a valve at transmission line with pressurized aerosol the time. As inhalator or insufflator, for example, gelatine capsule or soft cylinder can be by multivalence active material and the suitable support powders of packing into, such as lactose or starch, mixture make.

Medicament can be made the form of carrying out parenteral by injection, such as (in batches) injection or continuous drip. Injection can be made unit dosage form, for example, in ampoule or multi-dose container, wherein adds anticorrisive agent. Said composition can be formed in the suspension in oiliness or the aqueous carrier, solution or emulsion, and contain some preparatons, and such as suspending agent, stabilizing agent and/or dispersant. Perhaps, active component is made powdery, with suitable carrier, such as aseptic apyrogeneity matter water, be mixed with together parenteral solution before using.

Medicament also can be made the rectum composition, such as suppository or delay enema, for example, contains suppository base commonly used, such as suppository or the delay enema of coconut oil or other type glycerine.

Medicament can also be made the warehousing composition except above-mentioned preparation. This durative action preparation can be by implanting (such as subcutaneous or intramuscular) or the administration of intramuscular injection mode. Therefore, active material can with suitable polymerization or hydrophobic material (such as the emulsion that in usable oils, forms) or ion exchange resin, preparation or so that the derivative of a little solubility to be arranged is if any the form preparation of the salt of a little solubility.

The whole body administration also can be implemented through mucous membrane or skin. For through mucous membrane or through percutaneous drug delivery, to mix the penetrating agent that is suitable for penetrating the obstacle that will be permeated in the preparation. These penetrating agents generally are known in the art, for example, comprise bile salt and fusidic acid derivatives for mucosal. In addition, detergent can be used to strengthen penetration capacity. Mucosal can be implemented by nose internal spraying or suppository mode. For topical, available oligomer of the present invention is made ointment, ointment, and gelatin or emulsifiable paste, these forms are normally known in the art.

It is large and can not be passive and promptly by in the example of hydrophobic membrane to present thing a multivalence, inject or suction than alimentary canal or more suitable through percutaneous drug delivery. These tissues that are drug delivery are relevant with the medicine based on protein. Present thing in multivalence and in the example in GI road, can use the suitable method of presenting known in the art.

If necessary, composition can be wired up, or be contained in the distributor, or make the medicine box that illustrates with administration. Composition can contain one or more UDs that contain active component. Packing can be that for example, metal or plastic foil are such as blister pack. Available packing or distributor with medication instruction for example, are used for packing or the distributor with medication instruction of the inventive method. Multivalence is presented the thing set

The invention still further relates to multivalence and present the thing set. These set are used to method of the present invention. Multivalence is presented the thing set and is contained the multivalence of combining more than and present thing. This set can have the multiple different multivalence that will realize the function that they will be realized to present thing.

The invention still further relates to preparation, for example contain the therapeutic composition that above-mentioned multivalence is presented thing set and pharmaceutically suitable carrier. This multivalence that contains is presented the preparation of thing set and can be designed to selected different multivalence and present the preparation that thing can be given their characteristic whole preparation.

It can be the polydispersion form that multivalence is presented the thing set. To use term " polydispersion body " in the following discussion and represent set, and still, should be appreciated that, it relates to other form that multivalence of the present invention is presented the thing set equally.

Multivalence is presented thing polydispersion body and is contained more than one different multivalence and present thing. The polydispersion body that different multivalence are presented thing can be the set that the different size multivalence is presented thing, the different and degree that is replaced by group A of lengths of frame different, and/or these factors of difference of group A/ attachment combination have caused size different. The exact position of group A adhesion also can be different. Each member brings different character for whole preparation in the set, and this is very favorable when treatment disease or indication. For example, the low-molecular-weight member in the set can be than HMW member more promptly near their effect target. The HMW member is owing to have long residence time and/or the long life-span is arranged in relevant portion at action site, so it has longer action time than low-molecular-weight member but meanwhile. These differences are conducive to treat disease or indication, because presenting thing, multivalence can have the multiple character that the multivalence of the same race that comprehensively surpasses respective numbers is presented the character of thing (for example, (or different) with prolonging action time of arriving at rapidly as the site being combined). The method for the treatment of disease or indication

The invention still further relates to the method for the treatment of disease or indication. The method comprises to the treatment patient uses disease or the indication that some group A occur with treatment. Presenting the conformal interfacial interaction of thing and target binding site B set or some multivalence by multivalence in patient body in an example presents conformal interfacial interaction that thing and target binding site B gather and realizes treatment to disease or indication. Conformal interfacial interaction causes treating the interior binding site B set of patient body or target binding site B array is covered. Promote in another example the multivalence for the treatment of to present thing and meet a cover standard. This standard is as follows:

-group A is functional group and plays drug effect, and it is used alone or with framework, and for example, framework is not have alone enough affinity but the compound that can amplify by result of the present invention;

-the presenting of group A that adhere on the framework provides the interactional attendant advantages of presenting to a plurality of binding site B with respect to separate base A; And

-this attendant advantages is collaborative advantage, and this advantage is larger than the attendant advantages that the set that is dispersed in the identical group A member in the homogeneous solution may provide.

The example of attendant advantages comprises the biologicak efficiency that selects low concentration group A that abundance is provided, and improves selectivity and raising biotic potential for target and non-target site. Multivalence is presented thing at least two attendant advantages is provided in preferred embodiment, and multivalence is presented thing at least three attendant advantages are provided in preferred example. These advantages can be selected from above-listed advantage, discussing and those advantages described in multivalence is presented the relevant pharmacokinetics of thing function and/or mechanism one joint below also can being selected from, for example, positive cooperativity, the entropy that improves combination with three-dimensional inhibition is provided.

For the method for the treatment of disease or indication, binding site B is so positioned so that the multivalence interaction can occur. For example, multivalence is presented the group A of thing and the interaction of binding site B can be intracellular, extracellular (as, on the cell surface or on extracellular matrix) or be positioned at cell membrane. Multivalence of the present invention is presented thing can the several functions that promotes treatment, they can be used to (for example adjust, raise or downward modulation) state or the situation relevant with polyvalency, for example, undesirable state (such as fertilization) or morbid state (as infecting). The present invention presents thing and also is used to the special biological event (such as the prevention of cell-pathogen adhesion) for disease or indication and is used for treating special disease state (pass the patient with control or reverse the speed that infects as infecting to be with pathogen). In addition, the present invention presents thing these has aspect ill multiple external (exvivo) to use in conditioning. Nurse one's health special biological event

Multivalence of the present invention is presented thing and be can be used in some instances, for example, and conditioning cell-cell interaction. Cell-the cell interaction of large number of biological process need both can promote also can suppress this interaction and the present invention presents thing. A special example relates to the combination of cell and other cell, for example, and neutrophil leucocyte and arterial endothelial cell. The neutrophil leucocyte that is suspended at first in the blood that flows rapidly by the multivalence interaction is adhered on the endothelial cell also near inflammation site (A.Varki, " Journal of Clinical Investigation (J.Clin. Invest.) ", 1997,99,158-162; J.B.Lowe, P.A.Ward, " Journal of Clinical Investigation ", 1997,99,822-6; M.A.Gimbrone, T.Nagel, J.N.Topper, " Journal of Clinical Investigation ", 1997,99,2062-70). Near inflammation shows that the neutrophil leucocyte of initial fast transport has adhered to the surface of endothelial cell, and (10-20 μ m/min) then slowly rolls; Endothelial cell is the cell of aligning in the blood vessel. (these select abnormal protein ground to be present in the surface of these cells by many E-and CD62P on the endothelial cell surface in this rolling; They are being introduced by cell factor during the inflammation) show that with neutrophils surface the interaction between the GL-PP (a kind of tetrose) of the sour Lewis X of saliva mediates (M.A.Gimbrone, T.Nagel, J.N.Topper, " Journal of Clinical Investigation ", 1997,99,2062-70). In addition, L-selects albumen (appearing at neutrophils surface) and saliva acid Lewis X (appearing on the endothelial cell) to interact. The interactional chemical valence of this cover changes the dynamics of meeting appreciable impact neutrophil leucocyte rehabilitation, and mechanics and single-minded character (L.L.Kiessling, N.L.Pohl, " chemistry and biology " (Chem.﹠ Biol.), 1996,3,71-77). Table 2 provides other example of cell-cell interaction.

The example of table 2 multivalence cell-cell interaction

Cell

1	Molecule on the cell 1	Cell 2	Molecule on the cell 2
1	Molecule on the cell 1	Cell 2	Molecule on the cell 2	Neutrophil leucocyte	L-selects albumen, CD62P	Endothelial cell	The saliva acid Le of sulfuration^X
Neutrophil leucocyte	The saliva acid Le of sulfuration^X	Endothelial cell	CD62L	Neutrophil leucocyte	L-selects albumen, CD62P	Endothelial cell	The saliva acid Le of sulfuration^X
Neutrophil leucocyte	The saliva acid Le of sulfuration^X	Endothelial cell	CD62L		Neutrophil adherence molecule (NCAM), such as L1, NCAM-H, CD24 and α 6-integrin
Neutrophil leucocyte	CD62L, cadherin	Neutrophil leucocyte	The saliva acid Le of sulfuration^X
Neutrophil leucocyte	CD62L, cadherin	Neutrophil leucocyte	The saliva acid Le of sulfuration^X	The T cell	φt cell receptor	Antigen presenting cell	Ajor histocompatibility compound (MHC)
	CD3		MHC	The T cell	φt cell receptor	Antigen presenting cell	Ajor histocompatibility compound (MHC)
	CD3		MHC		CD28
Blood platelet	Ia/Iia	Endothelial cell	Collagen		CD28
Blood platelet	Ia/Iia	Endothelial cell	Collagen	Blood platelet	IIb/IIIa	Blood platelet	IIb/IIIa
Tumour cell	GalTase	Endothelial cell	GlcNAc	Blood platelet	IIb/IIIa	Blood platelet	IIb/IIIa
Tumour cell	GalTase	Endothelial cell	GlcNAc	Sperm	GalTase	Ovum	GlcNAc
Old and feeble red blood cell	Take off sialoglycoprotein	Liver cell	C type agglutinin	Sperm	GalTase	Ovum	GlcNAc

Multivalence of the present invention is presented cell-cell interaction that thing can be used to suppress another kind of type in another example, i.e. adhesion. For example, the adhesion of leucocyte and endothelial cell is to be mediated by known adhesion molecule family as selecting albumen. Selecting albumen is transmembrane glycoprotein, and expressing at blood platelet be CD62P, and expressing at leucocyte (comprising neutrophil leucocyte and other lymphocyte) is L-selection albumen, and expression is E-and CD62P at endothelial cell. Select the rolling on the protein mediated interior cutaneous vessel wall to interact, a kind of cause at last the interaction that leucocyte exosmoses (people such as Kretzschmar, " tetrahedron " (Tetrahedron), 1995,51:13015). The present invention presents thing will be used to the adhesion event and the subsequence spare that suppress initial, for example, and leucocyte migration. Multivalence of the present invention is presented thing and be can be used to disturb neutrophil leucocyte-endothelial cell to interact, therefore, the state that capable of regulating is relevant with neutrophil adherence, such as inflammation, adult's respiratory distress syndrome (ivrds), rheumatic arthritis, septic shock and reperfusion injury.

Multivalence of the present invention is presented thing and be can be used to suppress blood platelet-blood platelet interaction in another example. Platelet aggregation plays a part outstanding in thrombosis or thromboembolic events. ADP, fibrin ferment, adrenaline or collagen excitant are exposed to the fibrinogen binding site on flat board (plate) the Protein G PIIb-IIIa. Little peptide such as RGD or HHLGGAKQAGDV (H12), contains fibrinogen identification primitive, and can barrier fibers proteinogen and these sites bonding. Be attached to the hidden epi-position of inducing peptide on the GPIIb-IIIa express at the GPIIb-IIIa compound (people such as Gawaz, " artery sclerosis thrombus Vascular Biology " (Arterioscler Thromb Vasc.Biol.), 1996,16:621). Then, discharge the granule of platelet; One of main glycoprotein particle is thrombospondin, and its performance is mediation platelet aggregation (Gawaz, above-mentioned quoted passage). Select group A to suppress to cause all interactions of platelet aggregation. In preferred embodiment, the interaction of GPIIb-IIIa and fibrinogen identification primitive is adjusted. People wish to lure into treatment patient hypocoagulability in some instances, and people wish to lure into patient's hypercoagulability in other example.

Multivalence of the present invention is presented the ability that thing can be used to reduce cancer metastasis in another example. Transfer comprises cancer cell from initial site isolation out, then invades contiguous tissue, and under second site arrangement. Multivalence of the present invention is presented thing and can be worked at this three phases. Cancer cell can be by being invaded lectin-mediated (Matrosovich.989.FEB 252:1,2:1-4 organ or tumour cell to the tropism of each organ; The people such as Beuth, " clinical trial of transfer " (Clin.Exp.Metastasis), 1988,6:115-20). For example, beta lactose base bundle be described to potential metastases inhibitor (people such as Dean, " and carbohydrate compound research (Carbohydrate Research), 1993,245:175). Therefore, mix with conditioning agents as these identification events of group A and can be made into multivalence and present thing.

The present invention presents thing and can be used to adjust infection in another example. The interactional initial step of most cells-pathogen will relate to adhesion. This point is also set up all viruses. An adhesion that example is influenza virus and bronchus endothelial cell of virus and cell surface adhesion. Adhere to the surface of bronchus endothelial cell as the first step influenza virus that infects. Adhesion is that (HA tightly wraps in the agglutinin of virus surface, about 2-4 part/100nm by hemagglutinin²Or the about 600-1200 of each virion) (SA, the end position on many glycoprotein is sugared, and is the surface that is arranged in target cell densely, about 50-200 part/100nm with sialic acid for a plurality of tripolymers²) a plurality of parts between interaction cause. Table 3 provides other example (wherein, P=protein of cell-Virus Interaction; S=sugar (glycoprotein or glycolipid)). There is ever-increasing virus to be found to utilize more than one interaction mode and target cell adhesion.

The example of part and acceptor on table 3 virus and the host cell surface

Dna virus	Disease	Molecule on the virus	Molecule on the host
Dna virus	Disease	Molecule on the virus	Molecule on the host	Papovaviridae Polyomavirus polyomavirus	Contain the DNA tumour virus	VP1	S:NeuAc (α 2,3) Gal (β 1,3) GalNAc S: sialyloligosaccharide
Adenoviridae Mastadenovirus human body adenovirus	Acute respiratory disease, pneumonia, gastroenteritis (also being animal vector useful in the gene therapy)	The residue 1-141 P of P:E3gp (fiber desmin): the RGD base of penton base albumen (being different from fibrin)	P:I level MHC P: penton base and β₂Integrin interacts	Papovaviridae Polyomavirus polyomavirus	Contain the DNA tumour virus	VP1	S:NeuAc (α 2,3) Gal (β 1,3) GalNAc S: sialyloligosaccharide
Adenoviridae Mastadenovirus human body adenovirus			P:I level MHC P: penton base and β₂Integrin interacts	Herpesviral-α section Simplexvirus 1 type	Herpes simplex	P: glycoprotein E, D	P:gD, gH (the 2 steps adhesion of suggestion)
Varicella virus belongs to	Varicella zoster	Glycoprotein I (gE) (the gE dimer is similar to the Fc acceptor)		Herpesviral-α section Simplexvirus 1 type	Herpes simplex	P: glycoprotein E, D	P:gD, gH (the 2 steps adhesion of suggestion)
Varicella virus belongs to	Varicella zoster			Herpesviral-β section cytomegalovirus belongs to the human body cytomegalovirus	Blind and the slow main inducing of baby; Relevant with cancer (Kaposis sarcoma) with AIDS	P:gC-11,86kDa gp residue is similar to the 204-297 in H301 α-domain gene of I level MHC α 3	S: Heparan sulfate P:I level HLAMHC to β₂-microglobulin
Lymphocyte virus belongs to Epstein-Barr virus	Monocytosis,mononucleosis	P：gp350 P：gp3	The lymphocytic C3d acceptor of P:B type DR2 (CD21) is similar to C3 complement segment C3d				S: Heparan sulfate P:I level HLAMHC to β₂-microglobulin
Lymphocyte virus belongs to Epstein-Barr virus	Monocytosis,mononucleosis	P：gp350 P：gp3		Poxviridae orthopoxvirus vaccinia virus	Be used for little orthopoxvirus vaccine inoculation; Comparing with variola virus is very similar antigen partner	The residue 71-80 of P:VGF albumen is similar to EGF α	P: EGF-R ELISA. But some evidence is just in time opposite.
The Parvoviridae parvovirus belongs to the canine tooth parvovirus	This virus is the model for the people, and it causes erythema infectious disease (" the 5th disease "); If infected at period of gestation, then people's form can cause hydrops fetalis or cause death	P:GP1-anchor formula albumen	P:3201 T cell P: the surface of red blood cell precursor in the marrow (can cause anaemia after the infection)	Poxviridae orthopoxvirus vaccinia virus		The residue 71-80 of P:VGF albumen is similar to EGF α	P: EGF-R ELISA. But some evidence is just in time opposite.
		P:GP1-anchor formula albumen		The Hepadnaviridae viral hepatitis type b belongs to human hepatitis B virus	Serum hepatitis, chronic infection will cause patient's hepatonecrosis of 1-2%	The residue 21-47 P of P:Env albumen PreS part P:Env albumen PreS part: the PreS1 sequence of large S albumen	The P:IL-6 acceptor, Pre-S1 is in conjunction with albumen P: for the sero-abluminous liver cell acceptor P of polymerization: for the liver cell acceptor P of polymerization IgA: sialoglycoprotein P: the mediation sticking action runs through solubility IL6, obtains directly to enter non-hepatocellular ability P: the sialoglycoprotein acceptor
The fowl hepatitis B virus	The serum hepatitis of birds		P: glycoprotein 180

Table 3 (continued)

RNA virus
RNA virus				Picornaviridae enterovirus genus poliovirus	Polio	The residue 95-105 of P:VP1 capsid protein	P: the member of the super family of immunoglobulin (Ig) is not above-mentioned CD44
The Rhinovirus human rhinovirus	Common cold	P: line up the VP1 in a valley and the residue of VP3 master's capsid protein at virus surface	P：ICAM-1	Picornaviridae enterovirus genus poliovirus	Polio	The residue 95-105 of P:VP1 capsid protein
The Rhinovirus human rhinovirus	Common cold		P：ICAM-1	Cardiovirus encephalomyocarditis virus (Mongolia's heat)	Fever, rare causing death; Pathogenic former (for example, caused fatal myocarditis show effect with it pig) of many animals	The residue of P:VP1 and VP3 master's capsid protein	P: sialoglycoprotein P:VCAM-1
Aphtha (Aphthovirus) Tobamovirus foot and mouth disease virus	The highly infective disease of a kind of ox, sheep, pig and goat (the 10-70 % death rate); The people is caused death, but can be easy to be infected (causing high fever) by animal	The residue 145-147 (RGD) of P:NS28 glycoprotein P:VP1 albumen and residue 133-158 and the C petiolarea of 203-213 P:VP1 albumen	P: integrin	Cardiovirus encephalomyocarditis virus (Mongolia's heat)		The residue of P:VP1 and VP3 master's capsid protein	P: sialoglycoprotein P:VCAM-1
			P: integrin	Exhale intestines (arc) Viraceae rotavirus human rotavirus	Whole world human infant is suffered from a topmost inducement of gastroenteritis and diarrhoea, and 500,000,000 cases are arranged every year, and wherein 500 die ten thousand deaths and die	P: the C end parts of hemagglutinin	1.P:K562 erythroleukemia cell? 2.P: Beta-3 adrenergic receptor? 3.P: the sialoglycoprotein 4.S on the small intestine in the fine hair of cell: sialic acid
Togaviridae alphavirus Semliki forest virus	Fever, encephalitis	P: nucleocapsid	P:I level HLA and H-2MHC molecule	Exhale intestines (arc) Viraceae rotavirus human rotavirus		P: the C end parts of hemagglutinin
Togaviridae alphavirus Semliki forest virus	Fever, encephalitis	P: nucleocapsid	P:I level HLA and H-2MHC molecule	Lactic dehydrogenase-lifting virus	Fever, encephalitis	P: envelope glycoprotein (VP-3P)	P: the II 1a level MHC of macrophage
Sindbis virus	Relevant with Western equine encephalitis virus, the death rate is about 5%	P：E1-E2	P: high-affinity laminin	Lactic dehydrogenase-lifting virus	Fever, encephalitis	P: envelope glycoprotein (VP-3P)	P: the II 1a level MHC of macrophage
Sindbis virus		P：E1-E2	P: high-affinity laminin	Rubella (Rubivirus) Tobamovirus rubella virus	Rubella (German measles)		S: undetermined glycolipid^[57，58]
Orthomyxoviridae family Influenza Virus influenza virus	The main inducing of various forms of human respiratory system's diseases (comprising fatal pneumonia)	P: hemagglutinin (HN or H albumen)	P:HN albumen, liver cell S: (α 2 for sialyloligosaccharide influenza A S:NeuAc, 6) (β 1 for Gal, 4) (α 2 for GlcNAc S:NeuAc, 3) (β 1 for Gal, 3) GalNAc S:NeuAc (α 2,3) Gal (β Isosorbide-5-Nitrae) GlcNAc influenza C S:9-O-AcNeuAc	Rubella (Rubivirus) Tobamovirus rubella virus	Rubella (German measles)		S: undetermined glycolipid^[57，58]

Table 3 (continued)

Paramyxoviridae paramyxovirus genus sendai virus	The breathing syncytial viruses of upper (descending) respiratory tract infection of the only children of causing of the second as a kind of viral paramyxovirus	P: hemagglutinin	S: sialoglycoprotein S:NeuAc (α 2,3) Gal (β 1,3) GalNAc S:NeuAc (α 2,8) NeuAc (α 2,3) Gal (β 1,3) GalNAc S: gangliosides GDla
Paramyxoviridae paramyxovirus genus sendai virus		P: hemagglutinin		NDV	The infection of the upper respiratory tract	P: hemagglutinin-neuraminidase	S: sialoglycoprotein S:NeuAc (α 2,3) Gal (β 1,3) GalNAc
The Morbillivirus measles virus	Measles	P: measles virus hemagglutinin	P：CD46	NDV	The infection of the upper respiratory tract	P: hemagglutinin-neuraminidase	S: sialoglycoprotein S:NeuAc (α 2,3) Gal (β 1,3) GalNAc
The Morbillivirus measles virus	Measles	P: measles virus hemagglutinin	P：CD46	Pneumonitis virus belongs to Respiratory Syncytial Virus(RSV)	The inducement of the infection of the upper respiratory tract (bronchitis and pneumonia) may play effect, the main inducing of tympanitis aspect SIDS	P: fusion; Nucleocapsid
HCoV-OC43	The common cold pharyngitis		S：Neu5，9Ac ₂			P: fusion; Nucleocapsid
HCoV-OC43	The common cold pharyngitis		S：Neu5，9Ac ₂	The Rhabdoviridae Vesiculovirus belongs to vesicular stomatitis virus		P: glycoprotein	S: phosphatidylserine S: phosphatidylinositols S:GM3 gangliosides
The lyssavirus hydrophobin	Rabies	The residue 151-238 of P:G albumen is similar to curaremimetic neurotoxin	P:BHK-21 cell P: acetylcholinergic receptor (the residue 173-204 of the inferior unit of α) S: saliva acid gangliosides			P: glycoprotein
The lyssavirus hydrophobin	Rabies			The human C type Papovaviridae human T-leukemia virus (HTLV-1) of Retroviridae	Cancer---leukaemia	P: the residue 246-253 of envelope glycoprotein is similar to extracellular part and the IL2 of HLA-B MHC	P:I level HLA MHC molecule P: Interleukin 2 Receptor
Radiation leukemia virus	Cancer---leukaemia		P:T cell receptor: L3T4 molecular complex		Cancer---leukaemia		P:I level HLA MHC molecule P: Interleukin 2 Receptor
Radiation leukemia virus	Cancer---leukaemia		P:T cell receptor: L3T4 molecular complex	Amphotropic retrovirus	Cancer---leukaemia		P: the CD34 on the marrow progenitor cell
Avian type C oncovirus group avian leukosis virus	Cancer		P: LDL receptor	Amphotropic retrovirus	Cancer---leukaemia		P: the CD34 on the marrow progenitor cell
Avian type C oncovirus group avian leukosis virus	Cancer		P: LDL receptor	The mammalian type C oncovirus leukemia virus (Moloney) of being born in the year of mouse	Cancer---leukaemia	P: coating gp71	P: lymphocytic cell surface IgM P: 622 amino acid hydrophobins of unknown function
Parent's preferendum mouse retrovirus	Cancer	P: envelope glycoprotein	P: basic amino acid transport protein		Cancer---leukaemia	P: coating gp71
Parent's preferendum mouse retrovirus	Cancer	P: envelope glycoprotein	P: basic amino acid transport protein	Non-close preferendum murine leukemia virus	Leukaemia	P:gp70 (surface)	MHC

Table 3 (continued)

HIV-1	AIDS	P:gp120 is similar to Ig heavy chain district P:gp120, is similar to neural interleukin P:gp120, is similar to vasoactive intestines peptide	The CD4 P of P:T cell: with the CD4 molecule P of II level HLA-DRMHC interaction of molecules: cerebroside (or on the human colon epithelium HT29 cell closely-related molecule)
HIV-1	AIDS			HIV-2	AIDS		P：CD4
SIV	Be similar to the immunologic deficiency syndrome of monkey AIDS	P：SIV mac239	P：CD4	HIV-2	AIDS		P：CD4
SIV		P：SIV mac239	P：CD4	Africa swine disease poison		P：p12，p72

It is the first step of virus infections that virus is adhered on its host, and comprise on the virus surface a plurality of molecules simultaneously with its host cell surface on effect (the M. Tardieu of a plurality of molecules, R.L.Epstein, H.L.Weiner, " cytology Current Literature " (Int.Rev. Cytol.), 1982,80,27-61). When the adhesion process suppressed by the corresponding free monomer of this molecule of high concentration or by the monoclonal antibody of anti-this molecule, the molecule that is included in the mediation adhesion of virus and cell surface the most directly was identified. When part be sugar or the little minute period of the day from 11 p.m. to 1 a.m, this technology usually is used. Relate to the less direct characteristic of molecule of adhesion from the correlation that can not successfully be attached to the sudden change (both can be artificial, also can be natural) on the target cell; That is, if viral adhesion ability has been eliminated in the sudden change of Special Proteins, will relate to direct or indirect adhesion. Second technology usually is used for protein acceptor or part. Although there is the ever-increasing lip-deep molecule of quantity to be included in virus-cell recognition, usually be delphic. Because virion adheres to many materials, so it usually is very difficult wanting to tell abiotic combination from the biospecificity land nonspecificly. Some virions can enter host cell (cell uses same mechanism to remove the little molecule of internalization) by non-single-minded transhipment. Some albumen can be identified more than one part of cell surface on the virus surface. At last, some virion can contact with the albumen of free disperse in solution, next takes turns to the molecule of identifying cell (the mediation adhesion) surface. The molecule of almost all kinds on the virus bonding cell surface: carbohydrate (A.Varki, " glycobiology " (Glycobiol.), 1993,3,97-130) (for example, polyoma (H.Fried, L.D. Cahan, J.C.Paulson, " virology " (Virol.), 1981,109,188-192) and orthomyxovirus (J.C.Paulson, J.E.Sadler, R.L.Hill, " journal of biological chemistry " (J.Biol. Chem.), 1979,254,2120-24) identification sialyloligosaccharide); Phosphatidyl fat (for example, vesicular stomatitis virus (VSV) identification phosphatidylserine and phosphatidylinositols (P. Mastromarino, C.Conti, P.Goldoni, B.Hauttecoeur, N.Orsi, " general virology magazine " (J.Gen.Virol.), 1987,68,2359-69)); And albumen (for example, HIV identification CD4 (A.G.Dalgleish, P.C.L.Beverley, P.R.Clapham, D.H. Crawford, M.F.Greaves, R.A.Weiss, " nature (Nature) ", 1984,312,763-7), the human rhinovirus identifies adhesion molecule 1 (ICAM-1) (J.M.Greve, G. Davis in the cell, A.M.Meyer, C.P.Forte, S.C.Yost, C.W.Marlor, M.E.Kamarck, A.McClelland, " cell " (Cell), 1989,56,839-8470)).

The virus of having been studied extensively and profoundly in the multivalence part is adhered to and the inhibition system is orthomyxovirus A (X-31) (genetic engineering of influenza virus) (the W. J.Lees that is attached to the red blood cell surface, A.Spaltenstein, W.J.E.Kingery, G.M.Whitesides, " (pharmaceutical chemistry magazine " (J.Med.Chem.) 1994,37,3419-3433; M.Mammen, G. Dahmann, G, M.Whitesides, " pharmaceutical chemistry magazine " (J.Med.Chem.) 1995,38,4179-4190). Thisly adhere to that multi interaction produces by between viral HA and the cell SA time. Affinity between influenza virus and red blood cell is not also carried out Accurate Measurement so far.

Virus-the cell interaction of another broad research is the interaction between the albumen gp120 on the human immunodeficiency virus (HIV) and the 60K gC D4 on the T-cell surface. The large concentration soluble CD4 CD4 of ectochemistry cracking (from) suppresses to adhere to. HIV is can affect the nerves glioma and human rhabdomyosarcoma cells of the example that can be used for changing the virus of the molecule on the cell surface: HIV, all lacks CD4 on it. Dissolubility CD4 does not block this adhering to and these cell types infection (P.R. Clapham, J.N.Weber, D.Whitby subsequently, K.McIntosh, A.G.Dalgleish, P.J. Maddon, K.C.Deen, R.W.Sweet, R.A.Weiss, " nature " (Nature), 1989,305,60-62).

Have many about mediating the example of virus and Cell binding. An example relates to hepatitis B virus, and in some cases, it is in conjunction with the seralbumin of assembling; These are assembled conversely in conjunction with albuminous acceptor on the surface of hepatocytes (P.Pontisso, M.A.Petit, M.J. Bankowski, M.E.Peeples, " Journal of Virology " (J.Virol.) 1989,63,1981-1988). The example of second abundant research starts from the divalence of antiviral antibody and virus surface and adheres to (example comprises Dengue virus, West Nile virus, and HIV) (R.M.H. V.Van, G.Hardie, " immunochemistry " (immunochem.) 1976,13,503-507). Interact at the lip-deep Fc-acceptor of target cell and many Fc tail multivalence. The mechanism that macrophage in the identification of this mechanism and exotic disease substance uses is identical: what is interesting is that the former affects infection, the latter affects removing.

Bacterial pathogens can be divided into two classes: bacterial pathogens in the cell (enter cell and at those of that breeding) and extracellular bacterial pathogens (be grown in the cell, still do not enter those of cell interior). The mechanism that infects by bacterial pathogens in the cell is similar to viral mechanism: infect (table 4-5) by causing adhering to of pathogen and host cell. Extracellular bacterium often moves and is gathered in particular organization. Bacterial concentration in the particular organization (tissue tropism) can produce by the specific interaction between the special composition of the extracellular matrix of the molecule on the cell surface in the molecule on the bacterium surface and the preferred tissue or this tissue. Many specific interaction between cell and host cell or the extracellular matrix are in the past by summary (I.Ofek, N.Sharon, " the up-to-date topic of microbial immunology " (Curr. Topics Microbiol.Immunol.) 1990,151,90-113; Ofek, N.Sharon, " infection immunity " (Infect.Immun 1988,56,539-547). For virus, interactional specificity is defined by best inhibitor molecules or the CA of bonding usually. Be planting on agglutinin and the host cell or usually use this mensuration (table 4) during the interaction of the sugar on the extracellular matrix on the bacterium surface when interacting. Now, the interaction of bacterium seems than relating to not only as part but also as the interaction more obvious (table 5) of the virus of the albumen of acceptor. Interaction between these protein-proteins more is difficult to research, does not both know mainly due to " monomer ", can not obtain for research again, when perhaps taking out from film, loses its " shape ". As a result, so far, bacterium studies the interaction that relates to agglutinin-sugar in great detail with the interactional great majority of virus, rather than the interaction of protein-protein.

Enterobacteria is the bacterium that colonizes in the mammal intestines and stomach. In most of the cases, enterobacteria is expressed 1 type pili, and it is in conjunction with the mannose on the surface epithelial cell in the gutstring (for example, Escherichia coli, bacillus canalis capsulatus, and salmonella). This interaction can be measured by the guinea pig erythrocyte agglutination (with the surface of the dense parcel of mannose residue). Fully the example of research is the research to the urine pathogenic escherichia coli.

Some other intestines Escherichia coli contains the pili hemagglutinin in its surface, and this surface specific contains sialic glycoprotein in conjunction with various on the endothelial cell surface. A sialic acid identification example of fully studying occurs in (M. Lindahl on the Escherichia coli that contain K99 pili agglutinin, R.Brossmer, T.Wadstrom, " sugar in conjunction with magazine " (Glycoconji.J.), 1987,4,51-58), its identification specifically is positioned at the N-glycosyl neuraminic acid on NeuAC (α 2,3) Gal (β 1)-ceramide. The intestines bacterial strain of comma bacillus produces various hemagglutinin, and (R.A. Finkelstein, L.F.Hanne, " infection immunity " (Infect.Immun.) 1982,36,1199-1208). These hemagglutinin are the most general in conjunction with the fucose derivative on the inner skin surface.

Host organisms often starts the namely relative nonspecific defense of anti-these multivalence pathogen of multivalence. An example is the abundantest glycoprotein-Uromucoid during the people urinates. This glycoprotein contains the oligomerization mannose unit that the variable N-of quantity connects, and in conjunction with many bacterium (F.DellOlio, F.J.J.de Kanter, D.H.van den Eijnden, F. Serafini-Cessi, " carbohydrate compound research " (Carbohyd, Res.1988,178,327-332).

Most bacteriums infect also and to cause (2:1-4 for Matrosovich.1989, FEBS 252:1) by relating to the adhesion step that the carbohydrate that exists on bacterial adherence and the host cell determines bunch. An example of bacterial adherence cell surface relates to Escherichia coli and urethra endothelial cell. The urine pathogen bacterial strain of bacteria Escherichia coli interacts directly and indirect surface (accompanying drawing 2) in conjunction with endothelial cell in urethra and the bladder with multivalence. It is preferred to have identified that several bacterioproteins give this tissue. Two examples are P-pili (containing G albumen) and I type pili (containing the FimH adhesin), and they all are positioned at the Escherichia coli surface. We use the as an example explanation in this system of P-pili. At first, these urine pathogen bacteriums can be positioned on its P-pili silk end the agglutinoid Protein G with effectively and specifically adhesion have urethra, multiple the copying of the Gal of the glycolipid on the endothelial cell surface (β Isosorbide-5-Nitrae) Gal (pK antigen) part in the kidney particularly. The second, the multiple multivalence that copies of the lip-deep F-albumen of Escherichia coli is in conjunction with fibronectin, a kind of dissolved sugar albumen. Fibronectin conversely multivalence in conjunction with endothelial cell surface. Bacillus coli multiple is collected in these tissues of urethra, therein the breeding and may cause disease (particularly pyelonephritis). Usually, bacterium is both directly in conjunction with cell surface, also in conjunction with the molecule in the extracellular matrix of preferred tissue. Bacterium both in conjunction with sugar also in conjunction with albumen.

Table 4

Example in conjunction with the bacterium of the sugar on the host cell surface (S) and albumen (P)

Bacteria name	Disease	Molecule on the bacterium	Molecule on the host
Bacteria name	Disease	Molecule on the bacterium	Molecule on the host	Actinomyces naeslundii 12104 and actinomyces viscosus LY7	Chronic mouth disease		S: contain the glucose phosphatide (GSLs) of GalNAc-β, for example GalNAc (β 1,3) Gal (α) O-ethyl
Candida albicans	Mouthful and the intact animal fauna part of (female) genitals passage, cause the disease (the prolonged stay catheter is eliminated the faunistic antimicrobial of normal bacteria for diabetes for example, immune deficiency) of Secondary cases procatarxis symptom	P:37-kDa laminin receptor P:58-kDa is in conjunction with the Mannoproteins P:C3d acceptor P of fibrinogen: ubiquitin	S: the glucoside that contains fucose at inner skin surface	Actinomyces naeslundii 12104 and actinomyces viscosus LY7	Chronic mouth disease
Candida albicans			S: the glucoside that contains fucose at inner skin surface	The sand holes Chlamydia	Can cause the chronic keratoconjunctivitis sexually transmitted disease respiratory tract infection of scar and blind	S: the polysaccharide that contains 7-9 mannose residue	P: mannose-binding protein
Intestines are assembled Escherichia coli (EAggEC)	Food poisoning	P: hemagglutinin	S: sialic acid	The sand holes Chlamydia		S: the polysaccharide that contains 7-9 mannose residue	P: mannose-binding protein
Intestines are assembled Escherichia coli (EAggEC)	Food poisoning	P: hemagglutinin	S: sialic acid	Urine pathogen Escherichia coli	Urinary tract infection	P:PapG adhesin P:FimH adhesin	S: the Gal on the glycolipid (α Isosorbide-5-Nitrae) Gal P?: hematuria platelet lysin Ia and Ib

Table 4 (continued)

Form the Escherichia coli of S-pili	The meningitis cerebrovascular complication		S:NeuGc (α 2,3) Gal and NeuAc (α 2,8) NeuAc NeuGc (α 2,3) Gal and NeuAc (α 2,8) NeuAc (being found in the endothelial cell of cerebral microvascular)
Form the Escherichia coli of S-pili	The meningitis cerebrovascular complication			Helicobacter pylori	Gastroduodenal ulcer, cancer of the stomach	P: hemagglutinin P: layer adhesion is in conjunction with albumen	S:Lewis (b) blood is antigen S:Neu5Ac (α 2,3) Gal (SA relies on bacterial strain)
Mycoplasma pneumoniae	Pneumonia	P: 40 on the hair tail and 90kDa albumen	S: long-chain saliva polysaccharide	Helicobacter pylori	Gastroduodenal ulcer, cancer of the stomach
Mycoplasma pneumoniae	Pneumonia	P: 40 on the hair tail and 90kDa albumen	S: long-chain saliva polysaccharide	Mycoplasma bovis	Pneumonia, genitals and urinary tract infection	P: the albumen of 40 and 90kDa on the hair tail	S: sialic acid residues and possibility sulfatide group
Neisseria meningitidis	Meningitis, acute adrenal insufficiency	P: hair (being that unusually hair contains two lactose bases-2,4-diacetylamino-2,4,6-three deoxyhexamethyloses)	P: CD66 P on endothelial cell and the neutrocyte: at the glycoprotein G of the lip-deep RSV of target cell (coinfection element)	Mycoplasma bovis	Pneumonia, genitals and urinary tract infection	P: the albumen of 40 and 90kDa on the hair tail	S: sialic acid residues and possibility sulfatide group
Neisseria meningitidis	Meningitis, acute adrenal insufficiency			Porphyromonas gingivalis	Oulitis		S：D-GalNAc ^[116]
Pseudomonas aeruginosa	The bacterium colony zone lacks natural defence (conduit), and causes disease in the immune deficiency host	P: hair adhesin	S: take off sialic acid-GM1 and the GalNAc (β Isosorbide-5-Nitrae) that takes off sialic acid-GM2 or GM1	Porphyromonas gingivalis	Oulitis		S：D-GalNAc ^[116]
Pseudomonas aeruginosa		P: hair adhesin		Rhizobiun lupini		The P:L-FBP	The S:L-fucose
Staphylococcus saprophyticus	Urinary tract infection	P: surperficial agglutinin	S: blood is A (end GalNAc)	Rhizobiun lupini		The P:L-FBP	The S:L-fucose
Staphylococcus saprophyticus	Urinary tract infection	P: surperficial agglutinin	S: blood is A (end GalNAc)	Streptococcus suis	Meningitis		S: be present in neural amino α more than 2 → 3-Nac-lactosaminoglycan of the Gal that Pl and Pk blood are antigen (α Isosorbide-5-Nitrae) Gal S:N-acetyl group; NeuNAc (α 2,3) Gal (β Isosorbide-5-Nitrae) Glc NAc
M+ group A streptococcus	Nasopharyngitis; The release of erythrogen toxin can cause when scarlet fever is worked as the secondary infection influenza virus and may occur, and finally cause fatal pneumonia in these situations		P:C3, mainly C3b and iC3b on PMN	Streptococcus suis	Meningitis
M+ group A streptococcus			P:C3, mainly C3b and iC3b on PMN	Streptococcus sanguis sobrinus streptococcus (oral cavity)	Normal flora part at the upper respiratory tract: defective may cause disease		S: the sialic acid of SGP
Yersinia enterocolitica	Severely subnormal pain and diarrhoea; May cause fatal sepsis in some situation		S: the Gal in the intestines mucoprotein, GalNAc, Lac	Streptococcus sanguis sobrinus streptococcus (oral cavity)			S: the sialic acid of SGP

Table 5.1 is had a liking for the various local Fusobacterium necrioohorus large intestine fusobacterium varium large intestine difficulties of yogurt bacillus in conjunction with the avette bacteroid large intestine of the bacterium example bacteria name target tissue bacteroides fragilis large intestine bacteroides vulgatus large intestine bacteroides distasonis large intestine bacteroides thetaiotaomicron large intestine yeast milk acidfast bacilli large intestine of glycolipid lactosyl ceramides porcine derivative and is debated clostridium large intestine clostridium botulinum large intestine propionibacterium granulosum skin, large intestine propionibacterium acnes skin is Roy Deng Shi Propionibacterium dairy produce actinomyces viscosus mouth actinomyces naeslundii mouth shigella dysenteriae large intestine shigella flexneri large intestine bacillus ceylonensis A large intestine salmonella typhimurium large intestine Escherichia coli intestines comma bacillus small intestine not, large intestine campylobacter jejuni intestinal flu haemophilus respiratory tract artificial tuberculosis yersinia genus intestinal plague Yersinia ruckeri intestines Diplococcus gonorrhoeae genital tract pseudomonas aeruginosa respiratory tract

Table 5.2 is at the bacterium example of extracellular matrix in conjunction with sugar (S) and albumen (P)

Bacteria name	Disease	Acceptor on the bacterium (part)	Part on the host matrix (acceptor)
Bacteria name	Disease	Acceptor on the bacterium (part)	Part on the host matrix (acceptor)	The conidium aspergillus fumigatus	Various local fungals infect		P: the D district of fibrin original molecule
Borrelia burgdorferi	Lyme disease		P: beta 2 integrin alpha IIb β 3 (glycoprotein IIB-IIa) (RGD)	The conidium aspergillus fumigatus	Various local fungals infect		P: the D district of fibrin original molecule
Borrelia burgdorferi	Lyme disease			The pertussis bordetella	Pertussis (pertussis)	P:Pertactin and silk agglutinin (FHA); Cell binding sequence (RGD)
Candida albicans	Mouthful and the intact animal fauna part of (female) genitals passage, cause the disease (the prolonged stay catheter is eliminated the faunistic antimicrobial of normal bacteria for diabetes for example, immune deficiency) of Secondary cases procatarxis symptom	P:37-kDa laminin receptor P:58-kDa is in conjunction with the Mannoproteins P:C3d acceptor P of fibrinogen: ubiquitin	S: the streptococcic cell wall polysaccharides S of gordonii during coinfection: contain β-1,2 and α-1,2 and connect	The pertussis bordetella	Pertussis (pertussis)
Candida albicans				Chlamydia psittaci	Can cause the chronic keratoconjunctivitis sexually transmitted disease respiratory tract infection of scar and blind	P: heparin-binding protein	Heparan Sulfates GAG and minimum chain length ten carbohydrates
Enterococcus faecalis	Diarrhoea		S: galactolipin, fucose, and mannosamine, but be not mannose	Chlamydia psittaci		P: heparin-binding protein
Enterococcus faecalis	Diarrhoea			Escherichia coli and S pili	Alimentary infection	P:S-pilin SfaA	S: suppress S by NeuAc (α 2,3) lactose: the glycolipid that contains terminal Gal (3SO4) β-1 residue
Haemophilus influenzae	The infection of the upper respiratory tract, often Secondary cases influenza virus	High-molecular-weight protein (HMW-1 and HMW-2)		Escherichia coli and S pili	Alimentary infection	P:S-pilin SfaA
Haemophilus influenzae		High-molecular-weight protein (HMW-1 and HMW-2)		Leishmania donovani	Protozoan parasite causes kala-azar (visceral leishmaniasis)	P: heparin-binding protein	S: heparin
Johne's bacillus	It or not main human pathogen		P: fibronectin	Leishmania donovani		P: heparin-binding protein	S: heparin
Johne's bacillus	It or not main human pathogen		P: fibronectin	Cow mycobacteria	Chronic granuloma infects, particularly in the lung	P: albumen 85B and p55 albumen	P: the collagen land of fibronectin, the glycoprotein that exists in the blood plasma

Table 5.2 (continued)

Bird extracellular mycobacterium	Common pathogens in AIDS patient		Laminin, collagen I, and fibronectin
Bird extracellular mycobacterium	Common pathogens in AIDS patient		Laminin, collagen I, and fibronectin	The fertile Salmonella of middle Prey	Periodontal infection	P: lactoferrin binding protein (newborn iron suppresses combination)	P: fibronectin, Collagen type I and IV type and laminin
Salmonella enteritidis	Severe diarrhea (common food from chicken and egg is poisoned)		Fibronectin	The fertile Salmonella of middle Prey	Periodontal infection		P: fibronectin, Collagen type I and IV type and laminin
Salmonella enteritidis			Fibronectin	Staphylococcus aureus	Be responsible for the main human pathogen of many infection. May cause meningitis, endocarditis and osteomyelitis	Three of P:38-residue D primitive are repeated P: fibronectin binding protein (galel A and ZZ-FnBP B) P:60kDa albumen P: collagen land (CBD) P of collagen adhesin: contain two or three collagen receptors that copy that 187 amino acid repeat primitive	P: terminal 29 kDa fragment (Fn29K) P of the N-of fibronectin: fibronectin P: bone sialoprotein (BSP) little (～80 kDa) integrin combination contains the bone matrix glycoprotein P of RGD: chondrigen P: collagen P: the N-end region of fibronectin (Heparin-binding district)
	Osteomyelitis and infectional arthritis	P: collagen adhesin P: fibrin original receptor (AF) fibronectin-in conjunction with albumen (FnBP) P: agnoprotein (60kDa)	Collagen (high degree of specificity and affinity) P: fibrinogen P: laminin (pepsin derive (P1) fragment) P: vitronectin	Staphylococcus aureus
	Osteomyelitis and infectional arthritis			Streptococcus saprophyticus	Urinary tract infection		S: the adhesion that is partly suppressed by mannose
Streptococcus	Many intestines and stomach and respiratory tract infection		Collagen type I	Streptococcus saprophyticus	Urinary tract infection		S: the adhesion that is partly suppressed by mannose
Streptococcus	Many intestines and stomach and respiratory tract infection		Collagen type I	A family streptococcus	Infectious endocarditis, glomerulonephritis, and rheumatic fever	P: lipoteichoicacid (LTA) and M albumen (suppressing combination by LTA) P: glyceraldehyde-3-phosphate dehydrogenase (GAPDH) P: albumen F P:PAM (relevant M albumen) P:M3 albumen	P: fibronectin P: plasminogen and fibrinolysin P: how in conjunction with the blood plasma fibrinogen, albumin and fibronectin

Table 5.2 (continued)

Ephritis (+) and ephritis (-) and A family streptococcus	Glomerulonephritis	P: ephritis plasmin binding protein (NPBP)	P: PLI's (by EACA blocking-up combination)
Ephritis (+) and ephritis (-) and A family streptococcus	Glomerulonephritis	P: ephritis plasmin binding protein (NPBP)	P: PLI's (by EACA blocking-up combination)	Bargen's streptococcus	Urinary tract infection, endocarditis	The lipoteichoicacid derivative
Weak streptococcus	Non-human pathogen	P: surface protein (about 200kDa)	The extracellular matrix of emiocytosis (ECM)	Bargen's streptococcus	Urinary tract infection, endocarditis	The lipoteichoicacid derivative
Weak streptococcus	Non-human pathogen	P: surface protein (about 200kDa)	The extracellular matrix of emiocytosis (ECM)	Streptococcus dysgalactiae	Pneumonia		Laminin, collagen I, II and IV type, fibronectin, and vitronectin
Streptococcus pyogenes	Be responsible for the main pathogens of many systematicness and local infection, and relevant with rear streptococcus amynologic disease	P: albumen F (the X-ray structure is known) P: streptococcus fibronectin binding protein (Sfb albumen, 37-amino acid sequence) P:9kDa grape aminoglycan-in conjunction with albumen (GAG-BP) P:M albumen	P: fibronectin P: human heart muscle basal layer	Streptococcus dysgalactiae	Pneumonia
Streptococcus pyogenes			P: fibronectin P: human heart muscle basal layer	Atypia Wei Yong Shi coccus	Mouthfeel is dyed		S: with some population streptococcus copolymerization collection (lactose can suppress and lactose-can not suppress)
Yersinia enterocolitica	Unusual pain and diarrhoea	P: memebrane protein YadA	P: cartilage derive human cell fibronectin and human plasma fibronectin	Atypia Wei Yong Shi coccus	Mouthfeel is dyed
Yersinia enterocolitica	Unusual pain and diarrhoea	P: memebrane protein YadA		Artificial tuberculosis yersinia genus	Unusual pain and diarrhoea	P: adhesin YadA	P: beta 1 integrin

Another example relates to the combination of cell and multivalent molecule, such as bacterium, and antibody and macrophage. All types of antibody-albumen form one of immune important family-have acceptor site how of equal value: two (IgD, IgE, IgG, IgA), four (IgA), six (IgA) or ten (IgM). Multivalence is in conjunction with the structure-antigen of these antibody recognition or other is on bacterium, virus, parasite, medicine, the part that exists on " non-from body " cell, or comprise that other structure of non-covalent complex does not exist-the ubiquitous feature of seemingly immunity identification usually in blood circulation. These interaction antigens suppress important course of infection (for example adventive body adhering to target cell) and promote to remove (filtering the removing foreign particles by macrophage and the degraded of immune composition or by kidney). Here multivalence effect refers to that to the high-affinity on the surface with repetition epi-position or adhesion, wherein epi-position is defined as the surface characteristics of nearly all invasion pathogen.

Mannose receptor (Fc acceptor) effect on the mannose residue on the antibody afterbody (Fc part) and macrophage (removing the important white blood corpuscle type of infected particle) surface. Single Fc part is seemingly too weak so that do not bring out the response of macrophage with the interaction of its acceptor; Free antibodies in the ie in solution (not compound) is activated macrophage neither, does not also activate the monospecific antibody in conjunction with the exotic disease substance fragment of degraded. Yet, in conjunction with the multispecific antibody on infected particle surface and the polyceptor forceful action on the Macrophage Surface, and obtain three layers of stable structure by the interactional interface of multivalence.

In addition, not only the multivalence in this system be provided in the identification of macrophage bacterium stability and specific may, and also strictly depend on thereafter the interaction of multivalence by the effect of macrophage. Multi interaction between macrophage and the coated antibody bacterium causes the interconnection on macrophage receptor surface, and the internal signal in the initiation macrophage causes its degraded with picked-up (engulfing) bacterium.

Other example on multivalent molecule attached cell surface provides in table 6 and 7.

The bacteriotoxic acceptor of table 6

Bacterium	Toxin	Part
Bacterium	Toxin	Part	Comma bacillus	Cholera toxin	(β 1 for S:GMl:Gal, 3) (β 1 for GalNAc, 4) ((α 2 for NeuAc, 3)) (β 1 for Gal, 4) Glc (β) ceramide S: (α 2 for different part NeuAc, 3) (β 1 for Gal, 3) ((α 2 for NeuAc for GalNAc (β), 3)) (β 1 for Gal, 4) (β 1 for Glc (β) ceramide Gal (β) GalNAc, 4) ((α 2 for NeuAc, 8)) (α 2 for NeuAc, 3) (β 1 for Gal, 4) (β 1 for Glc (β) ceramide GalNAc (β Isosorbide-5-Nitrae) Gal (β 1,3) GalNAc, 4) (α 2 for NeuAc, 3) (β 1 for Gal (β Isosorbide-5-Nitrae) Glc (β) ceramide Fuc (α 2,3) Gal, 3) (β 1 for GalNAc, 4) (β 1 for NeuAc (α 2,3) Gal (β Isosorbide-5-Nitrae) Glc (β) ceramide Gal, 3) (β 1 for GalNAc, 4) (R-NeuAc (α 2,3) Gal (β Isosorbide-5-Nitrae) Glc (β) ceramide R=fluorescein CH, rhodamine B, or DNP
Escherichia coli	Heat-labile toxin	S：GM1	Comma bacillus	Cholera toxin

Table 6 (continued)

Clostridium tetanus	Tetanus toxin	(β 1 for S:Gal, 3) (β 1 for GalNAc, 4) (β 1 for (NeuAc (α 2,8)) NeuAc (α 2,3) Gal, 4) Glc (β) ceramide S: (α 2 for different part NeuAc, 3) ((α 2 for NeuAc for Gal (β 1,3) GalNAc (β Isosorbide-5-Nitrae), 8)) (α 2 for NeuAc, 3) (α 2 for Gal (β Isosorbide-5-Nitrae) Glc (β) ceramide NeuAc (α 2,8) NeuAc, 3) (β 1 for Gal, 3) (α 2 for NeuAc for GalNAc (β Isosorbide-5-Nitrae) (NeuAc (α 2,8)), 3) Gal (β Isosorbide-5-Nitrae) Glc (β) ceramide
Clostridium tetanus	Tetanus toxin		Clostridium botulinum	Botulinal toxin A and E	S:NeuAc (α 2,8) NeuAc (α 2,3) Gal (β 1,3) GalNAc (β Isosorbide-5-Nitrae) (NeuAc (α 2,8)) NeuAc (α 2,3) Gal (β Isosorbide-5-Nitrae) Glc (β) ceramide
Clostridium botulinum	Botulinum toxin B, C, and F	S:NeuAc (α 2,3) Gal (β 1,3) GalNAc (β Isosorbide-5-Nitrae) (NeuAc (α 2,8)) NeuAc (α 2,3) Gal (β Isosorbide-5-Nitrae) Glc (β) ceramide	Clostridium botulinum	Botulinal toxin A and E
Clostridium botulinum	Botulinum toxin B, C, and F		Clostridium botulinum	Botulinum toxin B	S:Glc (β) ceramide
Bacillus aerogenes capsulatus	The δ toxin	S:GalNAc (β Isosorbide-5-Nitrae) NeuAc (α 2,3) Gal (β Isosorbide-5-Nitrae) Glc (β) ceramide	Clostridium botulinum	Botulinum toxin B	S:Glc (β) ceramide
Bacillus aerogenes capsulatus	The δ toxin		Difficulty is distinguished clostridium	Toxin A shiga-like toxin (SLT)-I and SLT-II/Iic	(α 1 for S:Gal, 3) (β 1 for Gal, 4) (β 1 for GalNAc, 3) Glc (β) ceramide S:Gal (α Isosorbide-5-Nitrae) Glc (β) (Pl disaccharides), (α 1 for Gal, 4) (β 1 for Gal, 4) GalNAc (β) (Pl trisaccharide), Gal (β Isosorbide-5-Nitrae) Glc (β) (Pk trisaccharide)
Shigella dysenteriae or Escherichia coli	Shiga toxin Vero toxin	S:Gal (α Isosorbide-5-Nitrae) Gal (β) ceramide S:Gal (α Isosorbide-5-Nitrae) Gal (β Isosorbide-5-Nitrae) Glc (β) ceramide	Difficulty is distinguished clostridium	Toxin A shiga-like toxin (SLT)-I and SLT-II/Iic
Shigella dysenteriae or Escherichia coli	Shiga toxin Vero toxin		The pertussis bordetella	Pertussis toxin	S：NeuAc(α2，6)Gal
Shigella dysenteriae	Dysentery toxin	S：GlcNAc(β1)	The pertussis bordetella	Pertussis toxin	S：NeuAc(α2，6)Gal

The crosslinked example as single conduction mechanism of table 7 surface receptor

Process	Receptor-ligand
Process	Receptor-ligand	Mast cell loses grain (allergy)	Allergen widowization IgE (P, N: N) on the mast cell surface
Cell Differentiation and growth (Developmental Biology: differentiation, migration, programmed cell death)	Fibroblast growth factor (FGF) dimer FGF acceptor. In forming the temporary transient FGF of multivalence and other HBGF, form the heparin effect. (P, N: N) EGF (EGF) dimer EGF acceptor. (NP) cell factor stem cell factor (SCF) the dimer Kit acceptor on stem cell surface. (P, 1: 2) TGF (TGF) dimer TGF acceptor. (NP) platelet derived growth factor (PSGF) dimer pdgf receptor. (P, 1: 2) single human growth hormone (HGH) (hGH) dimer hGH acceptor. (P, 1: 2) HGF (HGF) dimer c-Met acceptor. Heparin can provide the temporary transient HGF of multivalence (referring to top FGF). (P, N: N) (P, N: N) VEGF (VEGF) is in conjunction with vegf receptor Tit-1 for the EPO dimer acceptor on the red blood cell precursor. (NP) IL6 six aggressiveness IL6 acceptors. (P) the dimeric acceptor of IL4. (P, 1: 2) human macrophage clone-stimulating factor (M-CSF) dimer acceptor. (P, 1: 2) lactation hormone can dimerization lactogen acceptor (PRLR). (P, 1: 2) two unit price granulocytes-clone's stimulating factor (G-CSF) stimulates the dimerization of two G-CSF acceptors. (NP) haptens and multivalence X half can antigen receptor (the free multivalence modification that the X haptens decompresses. Multivalence X haptens does not have effect). (P, N: N)	Mast cell loses grain (allergy)
		Parasympathetic nerve response (from nervous system)	By the assorted dimerization (NP of the acetylcholinergic receptor of unknown mechanisms?)
Acrosome reaction ((biology of reproduction) in embryonated egg interacts	GlcNAc (P, N: N) on the GalTase-ovum surface on the seminal fluid	Parasympathetic nerve response (from nervous system)
		Peptide independent cloning expansion (immunology) interaction T-cell clone expansion of B-cell	Antibody-glycocalyx on the B-cell surface is at microorganism surface (P, N: N) its acceptor of alpha-interferon (IFN-α) dimer (P, N: N) t lymphocyte receptor CD28 and CTLA-4 in conjunction with and carry out oligomerization by molecule CD80 (B7-7) and the CD86 (B7-2) of stimulation on antigen presenting cell altogether
Chemotaxis of Bacteria	Aspartate dimerization bacterial receptor Tar. (NP)

P, 1: 2=two receptor surfaces of divalence part dimerization of mixing; P, N: N surface receptor of N=N-valency ligand receptor oligomerization; But the P=multivalence, but has unknown ligand-receptor stoichiometry; NP=can non--multivalence mechanism.

Multivalence is also important in preventing unwanted interaction, namely with the effect of multivalence inhibitor. Promote required interaction in that polyvalent antigen is powerful, it is at some unwanted biological interaction of prevention, and particularly self multivalence effect (multivalence is used for the antagonism multivalence here) also is a kind of useful strategy. For example, the liquid coatings in most mammal lungs contains several mucoproteins (albumen is presented polysaccharide and stopped) in sialic acid SA. These mucoproteins, particularly? the 2-macroglobulin can be in conjunction with influenza and other SA in conjunction with virus, and therefore suppressing it adheres to target cell.

In the selection of the group A that multivalence is presented, the group A of preferred use and the effect of pathogen particle in interacting with the host. In an example, the present invention presents thing and is used to block cell-bacterial interactions. In another example, the present invention presents thing and is used to block cell-fungi interaction. In another example, the present invention presents the interaction that thing is used to block cell-virus. Also in example, the present invention presents thing and is used to block cell-parasitic interaction. For example, the entamoeba historlytica sporadin is in conjunction with host's galactolipin (Gal) and GalNAc (GalNac) residue. (people such as Adler, 1995, " journal of biological chemistry " be 270:5164 (J.Biol.Chem.)). In some instances, for example, gene therapy is used, and promotes cell-pathogen, and for example the interaction of cell-virus may need.

In another example, can prepare multivalence of the present invention and present thing, it is regulated pathogen-extracellular matrix and interacts. For example, preventing that the interactional multivalence of bacterium or fungi and extracellular matrix from presenting thing can be suppressed. In another example, the present invention presents the interaction that thing is used to regulate cell-extracellular matrix.

In another example, can the interactional multivalence of structural accommodation pathogen-pathogen present thing. This thing of presenting will be used for the treatment of owing to use external device, the infection symptoms that causes such as prosthese and conduit, or the infection symptoms in the rupture process of external biological film.

In another example, can regulate the interaction of cell-toxin. For example, from the shiga toxin of shigella dysenteriae I type in conjunction with cell glycoprotein or have the glycolipid of galactobiose (Gal α 1-4Gal β) determinant. Can prepare multivalence of the present invention and present thing to prevent to block the interactional group A of any cell-toxin. In another example, can prepare and hemorrhagin toxin, the multivalence that the toxin that is for example produced by Croatalus viridis viridis reacts is presented thing.

In another example, the present invention presents thing and can be used to regulate the cell effect relevant with multivalence. For example, can be by in the interaction of the cell-cell that usually causes cytokine secretion, substituting the cell factor that the stimulator Cell regulate produces by cell. For example, be presented at L-on the tumour cell select protein ligands to stimulate unicellular and macrophage to produce TNF. Advised saliva Lewis x (sLe^X) play the HEV part that L-selects albumen, but native ligand may more complicated (Putz and Mannel, 1996, Sand.J. " immunology " is 44:556-564 (Immunol.)). Simulation is presented thing by the multivalence with group A of the tumour cell family of monocyte identification can be used to simulate this reaction. Equally, block the presenting thing and can be used for suppressing this reaction with group A of this reaction. In another example, wherein multivalence is presented the generation that other symptom relevant with multivalence that thing can be used comprises the T cell cytokine, mast cell and/or basocyte threshing, lymphocytic selection, and programmed cell death (Setedtsov and Seledtsova, 1995 of T or B cell, Biomed﹠Pharmacother, 1996,50:170), all these symptoms are all relevant with multivalence. For example with the treatment of multivalence relevant disease or indication

Except the target particular organisms process relevant with disease or indication, the professional and technical personnel can also design the multivalence of those processes of target specified disease or indication or proof patient existence and present thing. The present invention will comprise all groups that multivalence exists or the purposes of presenting the medicine that thing partly provides as multivalence of the present invention. Referring to, for example " Harrison internal drug principle " (Harrison ' s Principles of Internal Medicine), the 13 edition, Eds, the people such as T. R.Harrison, McGraw-Hill N.Y., NY; Goodman and Gilman ' s " treatment pharmacological basis " (Pharmacological Basis of Therapeutics), the 9th edition, Eds. Joel GG Hardman, Alfred Gilman, Lee L.Limbrid, New York.McGraw Hill 1995; With the Physician Desk Reference the 50th edition, 1997. Oradell New Jersey, Medical Economics Co. All the elements of these books are all incorporated by reference here. The instruction of diseases related or symptom and relevant possible the medicine for example content of group A are incorporated by reference more especially at this. For example, can design multivalence and present thing and regulate and to catch, comprise, interact as suppressing the host parasite, increase immunity or vaccination strategies, reduce pyemia or septic shock. More specifically, the present invention presents thing and is used in particular for treating the infection of the upper respiratory tract; Infectious endocarditis; Interior abdominal infection or abscess; Acute infectious diarrhea disease and food posioning; Sexually transmitted disease; The pelvis inflammation; Urinary tract infection and pyelonephritis; Infectional arthritis; Osteomyelitis and prosthetic joint infect; Skin, muscle and soft tissue infection; Injectable drug uses the infection of device, from bite, the infection of scratch, burn and environmental organism body; Dye with hospital source sexuality. Therefore, the present invention presents thing and will be used for the treatment of infection (for example, the pneumococcal infection that is caused by gram-positive bacteria, staphy lococcus infection, streptococcal infection, corynebacteria infects, Listera spp infects, lockjaw, clostridium botulinum infects, and anthrax), symptom (for example meningococcal infection that is caused by gramnegative bacterium, gonococcal infection, Moraxella and Kingella belong to infection, and hemophilus infects, Legionnella infects, pertussis, enterobacteria infects, and Pseudomonas infects, salmonella, Shigella, campylobacter infects, cholera and other vibriosis, undulant fever, yatobyo, Yersinia infects, carrion's disease and groin granuloma. The present invention presents thing also will be used for the treatment of nocardiasis, lumpy jawl clams, and the mixing anaerobic bacterium infects, and mycobacterium and helicobacter pylori belong to infection (for example, pulmonary tuberculosis, leprosy and the infection of greedy mycobacterium). Multivalence is presented thing can also be used to treat conveyor screw disease (for example, syphilis, treponematosis, leptospirosis, relapsing fever, bad nurse borreliosis (lyme borreliosis)). Equally, present thing with multivalence of the present invention and will be conducive to treat Dermacentroxenus, Mycoplasma and infect according to Ureaplasma.

It also is favourable presenting thing treatment virus infections with multivalence of the present invention. For example, also can treat dna virus (for example, herpes simplex, varicella zoster, Epstein-Barr virus, cytomegalovirus infection, poxvirus infection, parvovirus, and HPV) and RNA virus (for example, retrovirus, influenza virus, enterogastritis, enterovirus and reovirus, measles, rubella, mumps, rabies, rhabdovirus, with the agent of class marburg virus (marbury-like agent), arbovirus infection, and arenavirus infection).

In other example, the present invention presents thing can be used to treat fungal infection, for example, histoplasmosis, coccidioidomycosis and paracoccidioidomycosis, blastomycosis, cryptococcosis, candidiasis, aspergillosis, mucormycosis, and other. In other example, multivalence is presented thing can be used to treat protozoal infections (for example, amoeba parasitosis, malaria and babesiasis, leishmaniasis, trypanosomiasis, toxoplasma disease, Pneumocystis carinii, Giardiasis, Cryptosporidiosis, and trichomoniasis). In other example, the present invention presents thing can be used to treat helminth infection (for example, trichinosis is organized nematode, gutstring worm, filariasis, Loa loa, onchocercosis, drachunculiasis, snail fever and other fluke disease, or taeniasis). In other example, the present invention presents thing can be used to treat vermin (ecotparasite) infection.

In another other example, the present invention presents thing also can be used to regulate immune response, namely by adjusted and this replying of lower adjusting. Therefore, multivalence of the present invention is presented thing and will be used for the treatment of in the immunologic deficiency disease (no matter which kind of potential cause) and treatment autoimmunity disease and thus immune-mediated damage. In some instances, the present invention present thing will be for suppressing graft-rejection.

In other example, multivalence of the present invention is presented thing and also is used for the treatment of blood coagulation disorders and thrombosis and is used for anticoagulation, molten fibre and Antiplatelet therapy.

In other example, the present invention presents thing and also can be used for treating neoplasia. In the preferred embodiment, the present invention presents the transfer that thing is used for suppressing primary tumor.

Certain plants derive and the biological agent of the toxin of bacterium in initial process relate to them and adhere to the multivalence of cell surface, be i.e. interaction and combination (Lord, J.M. with a plurality of cell ligand time the by a plurality of toxoreceptors; Roberts, L.M.; Robertus, J.D. FASEB 1994,8,201-208; Montecucco, C.; Papini, E.; Schiavo, G. Experientia 1996,52,1026-10321 Kuziemko, G.M.; Stroh, M.; Stevens, R.C. " biochemistry " (Biochem.) 1996,35,6375-6384; Richardson, J.M.; Evans, P.D.; Homan, S.W.; Donohue-Rolfe, A.Nature Struct.Biol. 1997,4,190-193). In one approach, multivalence polymerization galactoside can be used to block be commonly referred to ricin from the cytotoxicity agglutinin of common castor bean and the adhesion of mammalian cell surface. These polymer are by stoping the interaction between the galactoside residue on ricin and the cell surface to be worked. The quantity of the receptor-ligand binding that relates in the ricin cytoadherence is few.

Ricin, common castor bean agglutinin (RCA₆₀，RCA ₁₂₀) belong to a class cytotoxicity agglutinin, be called ribosomes-inactivating protein (RIPs). A kind of ricin hypotype, RC₆₀(A ₁B ₁Covalent complex; MW～65kDa) formed by two subunits; A-chain--glycosides enzyme, the specific purine bases of its hydrolytic rupture rRNA; Agglutinin (Kuziemko, G.M. with B-chain-sugared combination; Stroh, M.; Stevens, R. C.Biochem.1996,35,6375-6384) also referring to Baenziger, J.U.; Feite, D. " journal of biological chemistry " (J, Biol.Chem.) 1979,254,9795-9799; Houston, L. L.; Dooley, T.P. " journal of biological chemistry " (J.Biol.Chem.) 1982,257, and 4147-4151; Rivera-Sagredo, A.; Solis, D.; Diaz-Maurino, T.; Jimenez-Barbero, J.; Martin-Lomas, M.Eur.J.Biochem.1991,197,217-228. The second ricin hypotype, RCA₁₂₀(2xA’ ₁B’ ₁ MW～130kDa) is aspect 26S Proteasome Structure and Function many and RCA₆₀Closely related; It is non-covalent in conjunction with A ' by two₁B’ ₁The unit forms (Endo, Y.; Mitsui, K.; Motiguki, M.; Tsurgi, K. " journal of biological chemistry " (J.Biol. Chem.) 1987,262,5908-5912). The inactivation of A-chain catalysis rRNA is the main cause of ricin toxicity. Yet, do not have the cell internalization effect of toxin that this effect can not occur; This internalization effect occurs through acceptor (B-chain)-mediation encytosis.

The powerful biosynthesis that suppresses the albumen in the mammalian cell of ricin. The molecular action of ricin needs the cell internalization effect, and it is from the adhesion of ricin and cell surface; Be combined in the time of a plurality of galactosides (Gal) group of the mostly individual lactoside-recognition sites of this strong toxin-cell adhesion by ricin and cell surface glycoprotein and glycolipid and occur.

There is a plurality of β on the cell surface-D-galactolipin in Gal-binding site (each B chain about 3 sites) the multivalence selective binding of ricin by the B-chain, and ((β-GalNAc)-terminal oligosaccharides is attached to cell surface (Roberts for β-Gal) and β-N-ACETYL-D-GALACTOSAMINE, L.M; Lamb, F.I.; Pappin, D.J.C.; Lord, J.M. " journal of biological chemistry " (J.Biol.Chem.) 1985,260, and 15682-15685) (quantity of the Gal binding site of each B-subunit is always disputable. RCA is pointed out in many researchs₆₀And RCA₁₂₀Have respectively～3 (＞2) and～6 (＞4) Gal-site).¹²⁵The dissociation constant Kd that the binding proof of the ricin of I-mark adheres to HRBC's ricin (with the B-chain that separates) is～0.1-0.01 μ M. When studying by various technology (equilibrium dialysis, fluorescence and NMR), the simple unit price analog of β-D-galactoside is to RCA₆₀And RCA₁₂₀Have quite low affinity (Kd～10 with the B-chain that separates³-10 ²μ M). As a result, can produce that multivalence of the present invention is presented thing so that ricin quite closely is incorporated into cell surface.

Ricin aspect structure and the mode of action many with bacteriotoxin (anthrax; Cholera, shigella dysenteriae; The Vero cytotoxin) and the cytotoxicity agglutinin (ribosome inactivating protein of other plant derivation; Abrin) closely related. In the exploitation of cancer therapy drug, these toxin are used as at present the cell toxicant composition of immunotoxin and study, and have been divided into the menace material in the biological competition.

Therefore, in other example, the present invention presents thing and is used for preventing ricin and erythrocytic adhesion. For this reason, multivalence is presented thing and can be used for blocking-up based on the ricin toxicity of the polyvalency of blocking-up ricin-cytoadherence. In this method, with blocking the first step that it enters cell with the multivalence polymerization inhibitor of a plurality of identical Ligand Competition that exists on the cell surface. Here discussed and be intended to suppress ricin (RCA₆₀And RCA₁₂₀) with the synthetic and biologicall test of the polymerization multivalent molecule of chicken red blood adhesion. This inhibitor is the derivative of polyacrylic acid and butadiene maleic acid; They exist a plurality of identical galactosides that contain side chain as amide group: pAA (Gal) and pBMA (Gal). One of inhibitor, pAA (Gal-β_O-L1; χ Gal=0.4) shown inhibition RCA₁₂₀Activity: K_i ^HAI=0.14 μ M (take each galactoside as basic calculation); The activity of this polymerization multivalence galactoside is about 1500 times of unit price Gal-β-OMe, is unit price Gal-β O-L₁NH ₂270 times. Polymerization multivalence inhibitor is being blocked by RCA₁₂₀Compare RCA in the hemoagglutination that (having about 6 Gal acceptor sites) induces₆₀(having about 3 Gal acceptor sites) has significant activity. In hemagglutination was used, the molecular basis of polymerization multivalence galactoside increased activity interacted owing to the Gal acceptor site of ricin and the multivalence of the part of the Gal on the polymer (i.e. a plurality of (sites), simultaneously). We can suppress ricin-cytoadherence by verified synthetic glycopolymers. Therefore, these polymer are the new synthetic antitoxins of a class. Although these polymer can by stop Gal part on ricin Gal-binding site and the cell surface in conjunction with the effect of simulating t antibody, they also may introduce new Inhibitory Mechanism: the occupying of the Gal-binding site that entropy has raise; Make the Gal binding site steric stabilization that does not occupy by surrounding polymer. These polymer are to RCA₁₂₀Comparison RCA₆₀Having more activated research explanation multivalence inhibitor exists the acceptor of less part-binding site to have more activity to the acceptor with a greater number part-binding site than those.

Summarize with in the polymerization polyad and the strategy of ricin and the flow process of ricin-cell interaction be shown in accompanying drawing 3, it relates to following content: (a) receptor-ligand mediates ricin to the absorption of mammalian cell: the acceptor site of ricin (B-chain) is combined with a plurality of, specific cell ligand (beta galactose glycosides) multivalence and causes toxin-cell to be combined closely; The adhesion of ricin and cell is formed in the initial step in its toxic action. (b) ricin toxicity mechanism: after toxin enters cell interior (through endocytosis), reduced the cracking of A-chain from the B-chain, monomer A-chain is the N-glucosides of the specific purine bases of catalyzing hydrolysis rRNA effectively; This destruction has disturbed protein biology synthetic. (c) enter cell by polymerization multivalence galactoside blocking-up ricin.

Therefore, the Gal-that synthesizes of the present invention presents the adhesion that polymer can prevent as inhibitor ricin and mammalian cell. The compound that these are common, or the derivative of these compounds has as therapeutic agent and the prospect that contact of protective agent antagonism with ricin.

It also is important that the multivalence of toxoreceptor and specific cells-surface ligand is combined in other situation. For example, abrin contains a plurality of acceptor sites in each lps molecule B-chain; This toxin is optionally in conjunction with galactoside residue (Lord, J.M.; Roberts, L.M.; Robertus, J.D.FASEB 1994,8,201-208; Sandvig, K.; Olsnes, S.; Pihl, A. " journal of biological chemistry " (J.Biol.Chem.) 1976,251,3977-3984). Cholera toxin utilizes pentamer B (sugar-combination) subunit, wherein ((β 1 for Gal for the GM1 pentasaccharides on each identification cell surface, 3) (β 1 for GalNAc, 4) ((α 2 for NeuAc, 3)) (β 1 for Gal, 4) Glc (β 1,1) ceramide) residue (Kuziemko, G.M.; Stroh, M.; Strevens, R.C. " biochemistry " (Biochem.) 1996,35 6375-6384), shiga toxin and verotoxin utilize five B-subunits in conjunction with the glycosyl sphingolipid (globoside that contains Gal simultaneously, (β 1 for Gal (α Isosorbide-5-Nitrae) Gal (β Isosorbide-5-Nitrae) Glc, 1) ceramide) (Richardson, J.M.; Evans, P. D.; Homans, S.W.; Donohue-Rolfe, A.Nature Struct.Biol.1997,4,190-193).

Therefore, the present invention find to be used for design pathogen (cell)-Cell binding (be characterised in that high price: about 100-1000 site be potential can be used for interactional) with the inhibitor of toxin-cell interaction (being characterised in that low price: namely relate to≤the 10Gal acceptor site).

Multivalence of the present invention is presented thing and can be used to suppress ovum-essence interaction so that suppress fertilization, for example, presents thing with multivalence and can disturb the acrosome reaction. Therefore, the present invention presents thing and also is used to contraception, for example, interact by suppressing essence-ovum, for example, before sperm and ovum interaction, by causing acrosome reaction, for example precocious activating reaction, Inhibit sperm and ovum combination.

Fertilization is initial by the coated kind-specific adhesion in the extracellular of sperm and ovum. Sperm-ovum interacts and causes that sperm carries out a series of acrosomes reactions (acrosome process): the exocytosis of the acrosome capsule that sperm and ovum be coated with appears in early reaction. This acrosome reaction finally causes the fusion of sperm and ovum serous coat, so that sperm nucleus enters the cytoplasm of ovum.

In the mouse sperm, the surface protein β-Isosorbide-5-Nitrae on the sperm serous coat-galactosyltransferase (Gal-transferase) is the GlcNAc residue (accompanying drawing 4) of the coated oolemma (ZP) of specific identification ovum. Referring to accompanying drawing 4, flow process has been described and has been used the polymerization polyad, and pAA (GlcNAc) brings out the acrosome reaction of sperm and the overview of Inhibit sperm-ovum combination. (a; b) the mouse sperm is mediated by receptor-ligand binding with the adhesion of ovum: terminal N-acetyl group gucosamine (GlaNAc) residue of the upper ZP3 glycoprotein of oolemma (ZP) that the Gal-transferase on the sperm head surface and ovum are coated: (a) Gal-transferase acceptor be combined the aggegation that causes the Gal-transferase with the multivalence of a plurality of GlcNAc parts of ZP3, and (b) then the process Cellular Signaling Transduction Mediated bring out the acrosome exocytosis of sperm. (c, d) proposed mechanism is for bringing out acrosome reaction and Inhibit sperm-ovum adhesion by the GlcNAc that exists a plurality of identical poly-(acrylic acid) to present as amide side chains or pAA (GlcNAc). The description of accompanying drawing 4 is not measured.

It is believed that in various abnormal shapes (isoforms) the ZP glycoprotein of oolemma that ZP3 is the glycoprotein ligand of the Gal-transferase of mediation essence-ovum adhesion. ZP3 contains many identical O-connection N-acetyl group gucosamine (GlcNAc) part, the serine of its covalently bound glycoprotein and threonine residues. From a plurality of surface Ga l-transferase acceptors of residual base cluster energy while of the poly-GlcNAc of a plurality of ZP3 in conjunction with the sperm head.

The acrosome reaction of sperm occurs by Cellular Signaling Transduction Mediated mechanism, by the activation triggers of the protein-coupled acceptor of some G-. The activation of G-protein receptor needs the agglomeration in space (accompanying drawing 4) of Gal-transferase. In the process of normal essence-feritilization of ovum, ZP3 Glycoprotein binding and aggegation Gal-transferase among the ZP that ovum is coated with: this aggegation is the interactional result of multivalence of a plurality of GlcNAc parts and the sperm surface Gal-transferase of ZP3.

The analogies of the multivalence GlcNAc of a GlcNAc--ZP3 of valency form of the present invention have caused the acrosome reaction (accompanying drawing 4) of sperm. The acrosome exocytosis of polymerization multivalence GlcNAc mouse sperm, so the external combination of sperm and ovum.

In a preferred embodiment, multivalence of the present invention is presented thing and can be combined with other contraceptive device and maybe can be used to prevent disease, is used to simultaneously contraception. For example, group A is used for the treatment of bleb, HIV, chlamydia, HPV, the part of gonorrhoea and syphilis.

In further example, the present invention presents thing and also is used for the treatment of or regulates heart and suffer a shock, angina pectoris, thrombosis in the circulatory system, heart failure, artery sclerosis, hypertension, apoplexy, ISR, the kidney failure of secretion erythropoietin(EPO), secretion property disease, allergic reaction, psoriasis, reaping hook cell anemia, mematopoetic system Secondary cases treatment (chemotherapy, drug side-effect) secondary disease or Other diseases (shifting autoimmune disease), kidney, prostate cancer, the cancer of the brain, lung cancer, thyroid cancer, colon cancer, cancer of the stomach, liver cancer, cancer of pancreas is to the tumour resistance of chemotherapy, the adult onset diabetes, growth defect, CNS damage, neurodegenerative disease, schizophrenia, antimigraine, anxiety, depressed, compulsive disorder, (but its disease that causes is unknown has the teiology that can accept the multivalence medicine, comprises the heating of unknown cause spontaneous disease, and MOF), disease of eye (glaucoma, myopia, retinal detachment, long sight), menopause, disease of pregnancy (hypertension, toxaemia, the material that diabetes are pregnant with helping foot, help the material of giving a birth, assist the medicament of presenting), anesthesia, fertility, wound healing, incontinence, skin disease (acne, dandruff, uticaria), Drug side reaction, and the reverse of drug toxicity. Further example comprises indication or the disease that relates to adjusting, described adjusting is for example to promote cell death, regulate, for example promote, suppress or increase metabolic pathway (for example passing through enzyme) or target special receptor type, seven yuan of transfer membranes of bunch shape (transmembrance) acceptor for example, or enzyme is (for example in the cell, the extracellular, or membrane-bound). The method that Analysis and Identification and test multivalence are presented thing

The present invention also provides the method that multivalence is presented thing of analyzing, and the method can be used for identifying the effectiveness that thing is presented in required useful group A or test. These class methods can be external or body in. In addition, analyzed in vitro method test multivalence be can design and thing adjusting A and the interactional ability of B presented, or A and B (for example interact the biological response that causes, cytoadherence is measured, CA, and platelet aggregation is measured, ELISA measures, and muscular contraction force mensuration, infectious mensuration, or lymph spread effect mensuration etc.). In-vitro screening

In some instances, the effectiveness that can use external test test multivalence to present thing is namely measured the interaction that multivalence is presented thing and target binding site B, or the interaction of inhibitor group C and binding site B. For example, present in the thing in screening and can use this mensuration, interaction (for example elutriation the test) (Charych of the lip-deep binding site B of group A multivalence of thing is presented in i.e. test, D. wait the people, " chemistry and biology " (Chem ﹠ Biol.) 1996,3,113-120).

In another exemplary embodiment, can use Capillary Electrophoresis (CE). CE only needs the conventional high analytical technology that splits of femto mole material. CE can drag according to the electric charge of molecule mixture (ion, little molecule, polymer, albumen, micelle) and hydrodynamics and be isolated. Be added in the buffer solution by the group A with various concentration, and by the flowability affects of binding site B of this concentration of monitoring to injection, can the accurate quantification group and the binding constant of binding site. This technology is called affinity Capillary Electrophoresis (ACE). For example, can measure the present invention with ACE and present thing to the affinity of the whole virus of expression binding site B. Show that also ACE forms the basis of various effective laboratory researches. When presenting the thing analysis, when particularly it was electrically charged, CE and GPC and light scattering all were useful.

Can also use surface plasma resonance spectrum (referring to, for example, Mrksich, M.; Deng the people, Langmair 1995,4383; Mrksich, the people such as M., " JACS " (J.Am. Chem.Soc.) 1995,117:12009; Sigal, the people such as G.B., " analytical chemistry " (Anal. Chem.) 1996,68:490). Used surface plasma resonance (SRR) research surface conjunction process.

Can also use model surface based on the self assembly individual layer (SAM ' s) to measure the present invention and present the thing molecule. The alkanethiol compound of SAM ' s on gold and silver is that another research absorbs, or occurs in the model system of other molecular process between the surface. (referring to, Mriksich for example, M.; Whiotesides, G.M., Ann.Rev.Biomol.Struct.1996,25:55; Whitesides, G.M. and Gorman, C.B. self assembly individual layer: the model that is used for organic surface chemistry; CRP Press:Boca Raton, 1995; Mrksich, the people such as M., " JACS " (J.Am.Chem.Soc.) 1995,117:12009; Sigal, G.B waits the people, " analytical chemistry " (Anal.Chem.) 1996,68:490; Lopez waits the people, " JACS " (J.Am.Chem.Soc.) 1993,115:5877).

In another example, can test aggegation, for example, use with the synthetic globule of pathogen or mixing with cells (being equivalent to aggegation suppresses to measure) and come the quantitative analysis multivalent molecule. Aggegation is that method and the 96 hole titer plate of the most convenient of detection cell-cell and pathogen-cells contacting are the instruments that is specially adapted to this mensuration. Automatically it also is possible carrying out this mensuration. For example, can make up the interactional especially globule of existence and pathogen or cell surface. The structure of this globule can three-dimensional enter the group with respect to background. In addition, with the suitable tie point of group, be considered to based on available crystal structure. In addition, it is also conceivable that by using and hang down the significance that non-specific interaction was eliminated or weakened to the polyethylene glycol background, and the significance of surface group density. Perhaps, target cell (or alternative target cell) can be used for some example. For example, can use influenza virus in conjunction with red blood cell, but not globule.

In addition, can measure the relative and absolute concentration on the globule that has group A and the surface that has binding site B, the two causes that mixture forms " glue " (being aggegation). Can use painted globule with visible easily. Can be structured in the derivative that contains different mol ratio group, group on the globule, or the multivalence material of complete different group (in the situation, inhibition can be depended on the stereoscopic stable effect in the back). Usually, the effectiveness of multivalence material will depend on the mol ratio of the polymer monomer unit that connects active group in the test. The mol ratio of expection maximum effectiveness is system-dependent.

Can change these identical multivalence materials so that various other group is attached on the available carrier site. Specificity and effectiveness that the structure of various " assisting " (for example A2-An) group and mol ratio can increase molecule. Therefore, a kind of of this raising may mechanism be by the lip-deep hydrophobic pocket of random position pathogen (or cell) with the short hydrophobic side chain that hangs down mol ratio. The critical nature that expection control polymer is renderd a service comprises: the mol ratio of active group, electric charge, hydrophobicity, persistence length, randomness, physical size, and the quantity of bound water molecule.

Can use and suppress to measure, as, measure molecule and prevent biological surface, for example, virus, in conjunction with another biological surface, for example test of the degree of cell. Can carry out like this: by the competitive binding of molecule and acceptor, and prevent that surface bound groups is combined with the identical combination site. An exemplary mensuration is that hemagglutination suppresses (HAI) mensuration, and it will be described in detail in additional embodiment and appendix. HAI measures in the solution of red blood cells molecules in inhibiting virus aggegation (colloid formations) as basic. Can record easily the effective lower bound of HAI is～1nM.

Another exemplary mensuration is that Optical Collision (OPTOCOL) is measured, and it is that adhesion after the coated microballoon collision of red blood cell of inhibition under the existence of inhibitor and virus is arranged is the basis. This is determined at the counter-rotating microscopically and carries out a pair of two kinds of collision things of each tweezers clamping with the collimating optics tweezers. OPTOCOL measure quantitatively＜＜effectiveness of activated inhibitor still during 1nM.

HAI and OPTOCOL measure the concentration that all can record the part establishment, i.e. group A and in conjunction with the concentration of the suppressed about half of the interaction between the B. In HAI measured, suppressing constant was KHA and KOPTOCOL. When measuring the inhibition constant, the numerical value that records with every kind of mensuration all is reliably, KHA～=KOPTOCOL. These two kinds are suppressed constant and all are called KINH.

OPTOCOL is based on parallel double optical tweezers operation biomone people such as (, 1996 " biochemistries " are 3:757 (Chem.Biol.)) Mammen. This technology can be studied between, single red blood cell and have interaction between the single microballoon of influenza virus for example. This technology is used in particular for unusual tightly coupled system and also can be used for the mechanism that the research multivalence suppresses.

Mostly preferred assay method is weighed the feature that is used for obtaining the multivalence group that required target plays an important role. For example, the effective multivalent molecule for aggegation multivalence binding site purpose may form by the long group that flexibly connects the thing connection by " n " (greater than 10) are individual. Therefore, this mensuration is necessarily reacted the final purpose of polyad.

There are many detections, evaluation and the quantitative interactional method of multivalence. Some assay methods may be the direct mensuration of affinity; Can extract interactional free energy from these affinity. Other method can be measured the feature of compound aggegation, and only one of them is interactional free energy. These further features can comprise degree of hydration, the ability on stereoscopic stable molecule or surface, and/or the ability of crosslinked multivalence acceptor.

For quantitative binding constant (namely obtaining binding constant) from thermodynamics, must measure the relative scale of (directly or indirectly) not compound and compound group. According to the stability (with respect to its life-span) of compound, can use different technology.

Can use agglutination test to measure the ability of multivalence group aggegation multivalence binding site (precipitation, gel-forming, aggegation). For example, in immune precipitation determination, multivalence present thing can precipitation surface on multivalence binding site B. Although presenting the compatibility of polyad in the thing precipitation ability in the mensuration multivalence is important, further feature also is important. For example, when low concentration, presenting thing may be not and a plurality of B (polyB) combination; In some optium concentration district, precipitate; And when high concentration, each binding site B is combined with a group A, no longer precipitates. In this example, its similar antibody precipitation reaction only depends on compatibility can not determine type of precipitation. In vivoassay

Can carry out in vivoassay, for example, measure multivalence and present thing to the result for the treatment of of Animal diseases or indication, for example, can measure anti-infective. But this mensuration is not limited to, and measures to prevent the interactional inhibitor of multivalence. For example, this molecule not only by blocking-up adhesion host receptor, reduces and infects speed; And can remove mechanism by blocking-up, reduce clearance rate. One is measured example is the phage-infect of bacterium. Chu, Y.-H waits the people, Acc. Chem.Res.1995,28,461-468.

Table 8 explanation can be used for the various analysis that screening multivalence of the present invention is presented the thing useful quality.

Table 8 is the interactional various technology of multivalence quantitatively

Can measure
Can measure						Technology	The system that this technology has successfully been used	Affinity	Suppress surface-surface interaction (affinity-plus three-dimensional stability)	Dynamics	Comment
The blood clotting ion suppresses to measure	1. the interactional inhibitor 2. of influenza-red blood cell interacts with the Ab of many bacterium surfaces	Nothing	Have	Nothing	Be widely used; Carry out easily; Be generally limited to suppress constant greater than nM	Technology	The system that this technology has successfully been used	Affinity		Dynamics	Comment

Table 8 (continued)

ELISA	The interactional inhibitor of influenza-red blood cell	Have	Nothing	Nothing	The multivalence material that needs complex sign; Be generally limited to dissociation constant greater than 10-50nM
ELISA	The interactional inhibitor of influenza-red blood cell	Have	Nothing	Nothing		Fluorescent activation cell Corter	Interact with the Ab of cell surface	Have	Have in principle	Nothing	The covalency variant that needs Ab; Depend on combination and the follow-up Quantitative Separation of combining form not
OPTCOL (with the Optical Collision of two optical tweezers)	The interactional inhibitor of influenza-red blood cell	Nothing	Have	Nothing	Can measure related symptoms on the physiology (controlling cell impact velocity, relative orientation and other factors by the user); Mensuration relates to individual cells and with the coated single microballoon of virion, measures the lower bound of inhibition constant less than 10^-4M	Fluorescent activation cell Corter	Interact with the Ab of cell surface	Have	Have in principle	Nothing
OPTCOL (with the Optical Collision of two optical tweezers)	The interactional inhibitor of influenza-red blood cell	Nothing	Have	Nothing		The compatibility Capillary Electrophoresis	Vancomycin dimer and dimer D-Ala-D-Ala interact	Have	Nothing	Nothing	This technology has prospect very much, and may expand and be implemented into other system
Surface plasma resonance	Ab combination with the synthetic surface that has different densities antigen (DNP, anti--the DNP system)	Have	Nothing	Have	(other technology in this table generally need to be less than 10 to need mg multivalence material²); Other correlation technique comprises acoustics dish mould and surface acoustic wave, and all are according to the little charge detection in the electric medium constant of near interface	The compatibility Capillary Electrophoresis	Vancomycin dimer and dimer D-Ala-D-Ala interact	Have	Nothing	Nothing
Surface plasma resonance		Have	Nothing	Have		Pipette, extract (Evan Evans)	Cell-cell	Have in principle	Have in principle	Nothing	Quantitatively additional cell surface between adhesion and separation period according to the energy that needs distortion; Implementing may difficulty
Shear flow (T Springer and other)	Neutrocyte and interaction from the non-covalent surface of deriving of different selection albumen	Have	Have in principle	Nothing	According under light microscope, calculating a large amount of processes (adhesion, non--adhesion); Measure the possibility (with the functional form of solution by the flow velocity on surface) of adhesion	Pipette, extract (Evan Evans)	Cell-cell	Have in principle	Have in principle	Nothing
Shear flow (T Springer and other)		Have	Have in principle	Nothing		Dissociating under gravity effect		Have	Nothing	Nothing	Measure to reduce the time on surface, and with adhesion intensity correlation
Light microscope counting agglutination (Barry Shur)	The interaction of sperm-ovum, these interactional inhibition.	Have in principle	Have	Nothing	According to a large amount of processes of counting after the vibration (combination, not combination); With the possibility of inhibitor concentration as the function mensuration adhesion; Be difficult to carry out	Dissociating under gravity effect		Have	Nothing	Nothing
Light microscope counting agglutination (Barry Shur)		Have in principle	Have	Nothing		AFM	Contain the surface of streptavidin and contain the biotin surface interaction	Have	Have in principle	Nothing	Highly sensitive (can measure the interaction of individual molecule-molecule); Instrument is expensive
Light scattering	Nothing	Have	Have in principle	Nothing	Need; Need the covalency of wisp to derive	AFM		Have	Have in principle	Nothing

External application

The external application that the present invention also provides multivalence to present thing, for example, material and the system of the passive protection that biology is threatened are used for component and new diagnosis and the recognition system of the system of depolluting. As mentioned above, in this application, design multivalence of the present invention and present infection or the poisoning that the final blocking-up of thing is applied object. In some instances, the present invention presents thing and can also comprise one or more antimetabolic products. For example, contain multivalence by use and present the protectiveness veil of thing passive protection is provided, this multivalence present thing can be effectively to infectious agents or poisonous gas and as protectiveness overcoat, nightwear, hospital with dressing and washing agent. In addition, multivalence of the present invention can be presented thing and be added to solution and method for the system of depolluting.

In the external application of another kind, multivalence of the present invention is presented thing and can be used for, for example, and in the elutriation mensuration or optical detecting based on Surface absorption. For this external application, many aspects will be suitable for using, and not need to consider the needs of body internal contact compatibility. The mechanism of action

Following mechanism is used for purpose being discussed and never in any form conduct restriction explanation of the present invention. And, will also be understood that the factor such as " the β factor " objective standard rather than the mechanisms of action of relating to more. In a preferred embodiment, multivalence of the present invention is presented thing the binding site B ultrasonic is crossed affinity to unit price A (monoA). In other preferred embodiment, multivalence of the present invention is presented thing and is presented A than unit price and have larger specificity. The minimizing that " specificity " refers to polyA and the non-specific interaction of binding site nonB is lacked than being combined with monoA of observing. In other preferred embodiment, multivalence of the present invention is presented thing and is just had biological effect than when the concentration that A was observed that exists with the unit price form is lower. Multivalence of the present invention is presented thing and can be undertaken by one or more mechanism of following description.

For interacting as a class multivalence, there is not acceptable nomenclature. Because there are many different modes that binding site B (for example n receptor site) can be interacted with group A (for example N part), because interactionally depend on its details in conjunction with the free energy brute force, may there be the nomenclature of simple general-purpose. Here usedly be distributed in two N part and interactions between the n receptor on the entity and will be called Nth-order multivalence interact (illustrating that the 3rd-order multivalence interacts). It produces in conjunction with free energy Δ G^poly _N。

Show single ligand moiety and single acceptor interactional average free energy partly in the multivalence interaction, Δ G^poly _avgEqual Δ G^poly _N/ N (formula 3). Free energy unit price acceptor-ligand interaction Δ G that changes^mono The independent part free energy of N unit price (independent acceptor and N unit price interact) changes N Δ G^mono The most useful part that relatively relates to equal number and acceptor in conjunction with free energy, and if the part and the acceptor (with the not commensurate of the equilibrium constant) that relate to varying number must take into full account. Formula 3-5 provides corresponding binding constant value, K.

{ΔG}_{avg}^{poly} = {ΔG}_{N}^{poly} / N

(formula 3) Δ G=RTln (K) (formula 4)

K_{N}^{poly} = {(K_{avg}^{poly})}^{N}

(formula 5) concertedness: measure α

Ligand moiety and interactional average free energy (the Δ G of acceptor part in multivalence interacts^poly _avg) can greater than, equal, or less than free energy (the Δ G in similar unit price interacts^mono) (formula 6-8). The below is the nomenclature of accepting in the biochemistry, and the multivalence of these types interacts will be called positive coorperativity (work in coordination with), non--collaborative (add and) or negatively work in coordination with (interference). General Definition α is collaborative degree following (Connors, K.A. Binging Constants:The measurement of Molecular Complex Stability; John Wiley ﹠ Sons:New York, 1987). The interactional order of multivalence is depended in α unit.

{ΔG}_{avg}^{poly} = α {ΔG}^{mono}

(formula 6)

{NΔG}_{avg}^{poly} = {ΔG}_{N}^{poly} = αNΔ G^{mono}

(formula 7)

K_{N}^{poly} = {(K_{avg}^{poly})}^{N} = {(K^{mono})}^{αN}

(formula 8a)

α = \log [\frac{\log (K_{N}^{poly})}{\log {(K^{mono})}^{N}}]

(formula 8b)

Biological concertedness is discussed in the prior art. (Kosshland, D.E.; Neet, K. " biochemistry year summary " (Annu.Rev.Biochem) 1968,37,359; Perlmutter-Hayman, B.Acc.Chem.Res.1986,19,90-96). The biological positive cooperativity system of the best of research does not relate to polyvalency. For example, four oxygen molecule (O₂) have concertedness with the combination of tetramer hemoglobin; That is: the free energy when second oxygen is in conjunction with hemoglobin is more favourable than the combination of first oxygen molecule ,-Δ G^poly _avg＞-ΔG ^mono _first This non--the multivalence system in collaborative degree α greater than 1 or less than 1 unit (unitless).

As a class, multivalence interact always can not be often fully or modestly quantitatively (interaction of positive coorperativity multivalence), even in a system clear and definite inference. The positive cooperativity of multivalence system is a part of the present invention.

An example of positive coorperativity multivalence interaction (α＞1) can be pentamer cholera toxin and GM1, the oligosaccharides part combination of gm1 gangliosidosis. Cholera toxin is by five subunit (AB₅) form. The combination of toxin and monomer GM1 (the GM1 of the ceramide part cracking from the cell surface) provides by purely improve the good case study of the positive coorperativity of combination from enthalpy.

The entropy of monomer GM1 derivative and pentamer cholera toxin the first cohesive process equals the afterwards entropy of each cohesive process. Therefore, result from the different of enthalpy in that all of the free energy of monomer GM1 and toxin combination are different. The people such as Schoen have carried out pentamer B5 and five independently calorimetric research (Schoen, the A. of the combination in solution of GM1 oligosaccharides unit; Freie, E. " biochemistry " (Biochemisty) 1989,28,5019-24). First part binding constant is than second binding constant little 4 times (by a factor of 4). Calculate from pure statistics, the first part binding constant expection than large 5/2 times of second binding constant (interact for Nth order multivalence, Ki, i binding constant equals (N+l+i)/i. So N=5, K₂/K ₁=(5/1) (4/2)=5/2). Therefore be the combination of rising enthalpy.

Reported that pentavalent B5 be combined in essence irreversible generation and tightr than the combination of pentamer cholera toxin and 5 monomer GM1 unit with the cell surface of dense covering GM1. Because interactional enthalpy is approximately identical. For monomer GM1 be fixed on lip-deep GM1, this difference that may return the entropy that is fixed in these two types of combinations in conjunction with the difference of Δ G: the affinity of multivalence acceptor can be improved greatly by the multiple spot adhesion. Because pentamer not by quantitatively, even if its concertedness degree has, also is unknown to the multivalence surface affinity.

The below is possible be interactional two examples of negative collaborative (disturbing α＜1); Namely do not resemble the sort of situation of positive coorperativity (hemoglobin and O₂), second part occurs be combined with second acceptor to be combined with first acceptor than first part and have disadvantageous free energy. First example is bivalent antibody (Ab) and find to be wrapped in biological surface (such as mammalian cell, virus, the solid carrier between the ELISA test period) or to be fixed on part combination on the polymeric matrices dense. (Tanaka T.; Suzuno, R.; Nakamura, K.; Kuwahara, A.; Takeo, K.Electrophoresis 1986,7,204-209). Usually, significantly different for the unit price binding constant of the antibody of less organic part, but 10⁵-10 ⁸M ^-1In the scope. For non-, or the positive coorperativity system, we expect K^bi ₂＜(K ^mono ₂)～(10 ⁵-10 ⁸) ²M ^-1 The people such as Karush find the affinity high 30 times (having separated chemically the wherein univalent antibody of two or more binding sites of the existence in native protein with zymetology) than corresponding unit price Ab for the divalence Ab of the surface antigen in Bacillus sp combination; Be K^bi ₂＝30K ^mono＜(K ^mono) ²M ^-1(Karulin, A.Y.; Dzantiev, B., B. " molecular immunology " (Molecular Immunology) 1990,27,965-971). Therefore, the combination here is negative collaborative.

The concertedness that defines in the traditional biological chemical sense for the multivalence system both not with neither narrative parameter, because it is for the unit price system. Therefore, need a kind of multivalence system of describing in conjunction with the experience mensuration that improves. By the expansion of concertedness parameter alpha, we are called newly and measure (metric)) β. Therefore, although multivalence interacts and greatly to be better than any one monomer on qualitative and to interact, this monomer interact still phase mutual interference (Lees, W.J.; Spaltenstein, A.; Kingery, W.J.E.; Whitesides, G.M. " pharmaceutical chemistry magazine " (J.Med.Chem.) 1994,37,3419-3433) or differ from one another (multivalent ligand during the affinity chromatography " adhesion " compatibility gel may represent this situation). Only have by Quantitative Comparison multivalence and unit price interaction and could establish synergitic character. During interacting, multivalence improves affinity: numerical value β

In many multivalence system, the quantity of many ligand-receptor interactions, N is unknown. For example, shown by the erythrocytic multivalence inhibitor of influenza virus aggegation (the side chain fragment that is ended at sialic acid (SA) by polyacrylamide skeleton and a certain proportion of terminal forms) and the interaction of the lip-deep a plurality of hemagglutinin of influenza virus (HA) acceptor site specificity. These molecules prevent influenza virus and neuron target cell interaction. With the polymer (streptavidin that in the afterwards step of measuring, is used for adhesion enzyme-conjugation as part) of the side chain that has SA group and a small amount of biotin group, can measure the combination of this polymerization multivalence inhibitor and virus surface with ELISA class determination method. The virus of surface conjunction is cultivated the amount of the polymer of the strepto-biotin protein by enzyme-connection-biotin interaction indirect determination combination with the polymer that contains SA of various concentration. Measurable amount is the concentration of SA in constructing in conjunction with thermoisopleth, contains the amount of SA on the polymer in conjunction with virus surface. All can not know exactly in conjunction with the SA gene number (N) that is incorporated into the HA acceptor site on viral amount of polymer and the polymer. The quantity (as the parameter of [SA]) of the polymer by measuring combination can analyzing polymers and the interaction of virus. Half-maximum in conjunction with the time, 1/K^ELISA=[SA], wherein K^ELISAEqual binding constant (formula 9). Because the unknown of the N value in multivalence interacts here, report that can not relevant concertedness (α). K in this example^monoValue is 5 * 10²M ^-1 For us, the K of best best inhibitor^ELISAValue is 10⁸M ^-120 That is: in [SA]=2 * 10^-3During M, monomer SA half a maximum combination, and in [SA]=10^-8During M, contain polymer half maximum of SA in conjunction with virus surface. Therefore, its α value no matter, the multivalence inhibitor may be useful.

Therefore define the ratio of two binding constants for improving factor-beta

K ^ELISA＝βK ^mono(formula 9)

Be conducive to the in question concertedness (α) that multivalence system affinity improves: the molecule with high β value is useful, and whether the interaction of no matter producing between them works in coordination with. N is unknown in any system, but the total amount of the multivalent molecule of combination is known, and β will be useful parameter. Only have known N value, just can know to calculate known Δ G^poly _N，K ^poly _NAnd the exact relationship between the β (formula 10,11).

{ΔG}_{N}^{poly} = Δ G^{mono} - RT \ln (Nβ)

(formula 10)

β = (K_{N}^{poly} / N) / K^{mono}

(formula 11)

Wherein group A is that the multivalence of the present invention of carbohydrate is presented the β value of thing usually preferably greater than or equal to about 10⁸ Wherein group A right and wrong-carbohydrate, particularly group A are that the multivalence of the present invention of non-native ligand is presented thing and usually had the β value more than or equal to about 10, are preferably greater than about 10², more preferably greater than 10⁴ Although in conjunction with acceptor may have polyvalency, the non-natural part is not multivalence usually. The interactional enthalpy of multivalence

ΔG ^poly _NBy enthalpy (Δ H^poly _N) and entropy (Δ S^poly _N) component composition (formula 12). As first approximation, Δ H^poly _NValue is the summation of the interactional enthalpy of N unit price, N Δ H^poly This value increases by other interaction around avtive spot or reduces.

Δ G_{N}^{poly} = {ΔH}_{N}^{poly} - {TΔS}_{N}^{poly}

The combination that (formula 12) enthalpy raises

In some cases, the combination with a part of certain enthalpy and an acceptor may cause that next part is combined with its acceptor, and this latter's combination obtains larger enthalpy; I.e. Δ H in this case^poly _NValue is than Δ H^monoValue more negative (more favourable). This combination is the rising of enthalpy. The combination that enthalpy drop is low

If a part disturbs next cohesive process in conjunction with its acceptor, then the interactional enthalpy of multivalence will be unfavorable for that N the unit price of equal value of expecting interacts. This combination is the reduction of enthalpy. The combination that enthalpy reduces may occur when a plurality of ligand-receptor interactions formation between two multivalence entities need disadvantageous molecular conformation from energy. Rule of thumb, conformation are got over the multivalence entity of rigidity, more may make part and acceptor Space mismatching thereof, the combination (unless the geometry of part and acceptor coupling is accurate) that this will cause enthalpy to reduce. Interactional entropy

Understand the interactional entropy of multivalence and be necessary to understand the correlation that unit price is combined with multivalence. In the design of multivalence inhibitor, not exclusively understand entropy and caused many synthetic multivalent molecules only more effective at the edge than its unit price homologue.

The interactional total entropy Δ of multivalence S^poly _NShould consider translational entropy (the Δ S in acceptor and the part cohesive process^poly _trans，N), rotation entropy (Δ S^poly _rot，N) and conformational entropy (Δ S^poly _conf，N) variation contribution, and around Entropy Changes (the Δ S of water^poly _H2O，N) contribution; Usually be because the entropy (formula 12) of hydrophobic interaction largely.

{ΔS}_{N}^{poly} = {ΔS}_{trans, N}^{poly} + {ΔS}_{rot, N}^{poly} + Δ S_{conf, N}^{poly} + {ΔS}_{H 2 O, N}^{poly}

(formula 12) translation and rotation entropy

Molecule translational entropy Δ S_transCome from it and pass through the independently freedom of translation of space; Δ S_transBe worth relevant with the logarithm of its quality, M (Δ S_trans∝ 1n (M)), with the opposite M of logarithm (the Δ S of its concentration_trans∝1n[L] ^-1), rotation entropy Δ S_rotCome from particle around the freedom of its all 3 main shafts rotations, and relevant with the logarithm of the generation of its 3 moments of inertia, I χ, I γ and Iz (Δ S_rot∝ ln (IxIyIz)). So Δ S of particle_transValue and Δ S_rotValue only weak (logarithm ground) relies on its quality and diameter. For first approximation, all particle-acceptors, part, the translation of acceptor-part agglutinator and rotation entropy all equate. When two particles in conjunction with the time, 3 kinds of translations and 3 kinds of rotation frees degree have all lost. If (quality of particle is through being everlasting within 10 times to ignore the mass discrepancy of particle, the overwhelming majority is within 100 times), total translation of two of combination particles is approximately equal with the rotation entropy so, and no matter they are unit price or multivalence, and condition is that their concentration is identical. In biology, the concentration of molecule can change in greater than 9 magnitudes (nM-pM). Even translational entropy only depends on concentration in logarithm ground, but this wide region is so that the concentration that people are familiar with interacting particles is necessary to the importance of estimating translational entropy: this value increases along with reducing of concentration. Conformational entropy

Entropy and enthalpy can the interactional affinity of partial-compensation multivalence impact: wherein the conformation flexibility increases the conformation entropy of combination, and identical flexibility increase does not have to occur under the thermodynamics tension force possibility of all ligand-receptor interactions.

Be difficult to quantitatively with this loss in the conformational entropy that the multivalence acceptor is combined at multivalent ligand as everyone knows. The Entropy Changes of the carbon-carbon bond of fixed single, rotation is about 0.5kcal/mol (ref). Scope for other single key is 0.1-1.0kcal/mol. If originally all lost the freedom of reversing of all many complexings in the cooperation process for the key that rotates freely, with the maximum loss of occurred conformation entropy. For this loss Δ S_confThe upper limit estimate to be about 0.5Nkcal/mol, wherein N be two parts or acceptor in conjunction with in the quantity of single key. For long flexible chain, this number may be very large: the triethylene glycol sept, its loss up to 10kcal/mol may be disadvantageous. The solvation entropy

In water system, be the Entropy Changes Δ S of hydrone on every side to the last contribution in conjunction with total entropy_H2O Be the release of the organic water on the surface that exposes of biomolecule to the contributor who interacts in the water (comprise hydrophobic interaction and relate to the interaction of polar group), cause the entropy increase. For the quantitative assay of hydrophobic interaction with predict extensive overview (Blokzijl, W.; Engberts, B.F.N.Angew.Chem.Int.Ed Engl.1993,32,1545-1579). These terms will be similar to each part in unit price and the multivalence system (unless attachment changes conformation or is combined with receptor surface, the result changes the combination of the solvent of itself and complexing) usually.

Permitted eurypalynous multivalence and interact, the feature of its entropy is significantly different. When two materials begin freely to spread (begin two particles 6 direction translations, be reduced to 3 direction translations after the complexing) in solution, may interact, we are categorized as 3-dimension (3D) with this interaction; When one or two material is limited in (being categorized as 2 dimensions, 2D) when spread on a plane; When being limited in a line diffusion, interactional material (is categorized as 1 dimension, 1D). In conjunction with translation and the rotation entropy depend in complexing translation and the rotation free degree loss. This value (Δ S_transValue and Δ S_rotThe value sum) maximum in particle 3D combination, take second place among the 2D, minimum among the 1D. That is: these materials that spread less than independence in conjunction with entropy between N the part (such as two parts that are limited in cell surface) and N the acceptor (such as the transfer membrane albumen on another cell surface) that also freely is diffused among the 2D that freely are diffused among the 2D freely disperse the interactional entropy in the 3D mode. Dynamics and raising affinity

In some instances, multivalence of the present invention is presented thing by the operation of extremely low dissociation rate (off-rates), so that they are the effectively combinations of " forever " with respect to the time range of the bioprocess for the treatment of. This dynamics difference between polyA and monoA, namely low dissociation rate of multivalence form is that the present invention presents another mechanism that thing is different from monoA.

The interactional dynamics research of high-affinity thinks that improving mainly is because the dissociation rate (k of two multivalence entities_off) reduce rather than the increase of association rate. The combination of anti--DNP Ab and DNP-lys is with respect to the surface conjunction of the Φ X174 of identical Ab and DNP-covering, the k of the mating surface of affirmation_onValue only is 2 times poor (k_on(surface)～3.7xM^-1 S ^-1，k _on(DNA-lys)～8×10 ⁷M ^-1s ^-1), but k_offValue differs 10⁴(k _off(surface)～3.3 * 10^-4M ^-1s ^-1，k _off(DNA-lys)～1.0s ^-1). Because these speed qualitative relevant with (usually quantitatively relevant and) its thermodynamics, (Agmon, N.; Levine, R.D.Chem. Phvs.Lett.1977,52,197-2201), and these mensuration are consistent with polyvalency instinctively: the dynamics value between first ligand-receptor interaction of two multivalence entities approximates the interactional thermodynamics value of similar unit price; Therefore, association rate may be similar seemingly rational. Dissociating of the interactional material of multivalence needs the interaction of N part-acceptor of fracture; Therefore, occur in multivalence interacts, interacting than the unit price of needs slower dissociate seemingly rational. The three-dimensional inhibition

In some instances, multivalence of the present invention is presented thing and is worked by " three-dimensional suppress ". The three-dimensional inhibition is new strategy in effective drug design. In the infectious agent situation, for example, can design the multivalence inhibitor that adheres to, it relates to any molecule on the infectious agent surface of combining closely, and the polymer of namely presenting molecule does not need directly to adhere to. For example, can by present with virus surface on the group of combination of neuraminidase (NA) make up the polymer prevent influenza virus and red cell adherence. Choi, S.-K.; Deng the people, " chemistry and biology) " (Chem. ﹠ Biol.) 1996,3,97-104. Those skilled in the art pay close attention to usually as the NA site that does not mediate adhesion, therefore, observe anti-stick connect effect, the possibility of result generation aggregation colloid layer and virus surface adhesion with direct polymer. This " effect " may be " pure " three-dimensional inhibition, and namely occupying of entropy raising do not appear in viral albumen commonly used in adhesion, the avtive spot of hemagglutinin. Therefore, the medicine multivalence is presented and may be changed pharmaceutically-active original mechanism. The example of this medicine is not also known at present.

It is believed that the three-dimensional mechanism that suppresses more is relevant to the collision stabilization, but not acceptor-mediated process, although it depends on the specificity of acceptor-domination, so that the suitable binding site B of polymer target. When bring together two parts or group one or two when coated by the gel layer, its from the entropy unfavorable (the conformation flowability of polymer is reduced because rise near another surface the time) and from the enthalpy also unfavorable (because being unfavorable for osmosis). It is unique when it works by this mechanism that thing is presented in polymerization, and (Kingery-Wood waits the people although there is other relevant mechanism to come across liposome, J.Am.Chem.Soc., 1992,114,7303-7305) and tree (dendrimers) (Roy, R.; Tropper, F.D. " chemistry can will " (J.Chem.Soc.), " chemical communication " (Chem.Commun.) 1988,1058-1060; Roy, the people such as R., " (JACS " (American Chemical Society): Wasshington, D.C., 1994). Absorption and precipitation

In some instances, multivalence of the present invention is presented thing and is worked by the absorption that mediates lip-deep binding site B. By this mechanism, binding site B may effectively remove from solution or remove by the patient, for example a kind of virion. In another example, multivalence present thing by the precipitation or aggegation work. Pharmacodynamics

Structure and the intrinsic property thereof selected will affect the pharmacodynamics that multivalence is presented thing. For example, when design multivalence of the present invention was presented thing, two properties of consideration were dissolubility and size. Dissolubility

In most examples, multivalence of the present invention is presented thing and is had more water-soluble (for example in the mg/ml scope or in the higher scope) than conventional medicine. Dissolubility (and the following stated size) may affect the pharmacodynamics that multivalence is presented thing. The dissolubility of for example, presenting thing may affect following one or more relevant characteristic.

For example, dissolubility can affect the removing feature that multivalence of the present invention is presented thing. Scavenging action may increase suddenly when the dissolubility of molecule increases. Kidney tendency filtered water dissolubility molecule is faster. Equally, the medicine clearance rate directly and administration frequency proportional.

The water-soluble that the present invention presents thing can also affect the action period of presenting thing. In preferred embodiment, the present invention presents the A that thing presents than unit price and has longer action period.

In other example, the dissolubility of polyA may affect therapeutic index. Here used term " therapeutic index " refers to (LD50/ED50), and it can be measured according to methods known in the art. Here used therapeutic index refers to calculate on each group A basis. Therapeutic index and administration frequency are inversely proportional to. Because the present invention presents the low clearance rate of thing, polyA can deliver medicine to the patient with lower frequency than monoA. And, polyA will show that than monoA lower concentration changes in the patient in time, because the clearance rate of these medicines may be very slow, they in addition can remain on the PC level, that is: the multivalence medicine will reduce the minimum and maximum concentration difference in site interested (for example reducing paddy-peak difference). Because multivalent molecule is large, they have significant advantage, and its life-span significantly lengthens than little molecule. Because the present invention presents the lower clearance rate of thing, polyA can be than monoA with the low frequency dosed administration with the patient. This long half-life is because many-sided former thereby become favourable. For example: (i) increase patient's obedience and happiness owing to reducing administration frequency; (ii) patient is comparable more early leaves hospital now; (iii) medicine can be than existing low treatment index administration.

Multivalence of the present invention is presented the thing dissolubility can also affect the chamber that multivalence is presented thing. Except dissolubility, other factors also may affect chamber, and the form of ownership of chamber for example comprises or gets rid of and will belong to a part of the present invention. For example, the breadth coefficient of water and organic solvent (simulation biotic environment) is important. Polymerization multivalence material with HMW generally can not effectively pass through biomembrane. In some instances, this feature is so that preferably directly be it and be handed to interested chamber administration. Perhaps, this character refers to that multivalence presents thing and can be excluded outside undesirable chamber. As an example, intravenous injection intravasation zone; Intrathecal injection enters myelencephalon liquid and central nervous system; The oral intestines and stomach that enter; Eye drip enters the eyes zone, and emulsion and ointment enter epithelium; Conduit enters biliary ductal tree and pancreas and gall-bladder, and bladder urethra system, and vagina fallopian tubal ovary system; Suction enters the bronchus zone.

Can also design such molecule to remain on outside the specific chamber. Wherein the example in this adaptable zone of conception of species comprises obstetrics' (keeping medicine not enter in neonate's circulation), to the virose medicine of kidney (in circulation, avoiding it to be absorbed by kidney by keeping medicine), or avoid presenting thing and enter in the central nervous system.

Multivalence is presented thing and can also be had tendency location and be retained in interested site, and this advantage has reduced system toxicity and maximization local concentration.

In addition, can design such molecule, wish that when certain character it is not limited to certain zone. For example, in case of emergency, can not wish that it is long-acting medicine, but importantly the multivalence medicine can increase effectiveness. In these are used, can be median size with polymer design. Perhaps can comprise dissociable attachment. These molecules will be effectively but enough little so that filter and removing by kidney.

Some examples of specific chamber comprise eye (excitant and antagonist or the antibiotic of shedding tears at during surgery), GI road (for example wriggle excitant and antagonist) (cholinergic excitant) (cathartic) excitant and muscular tone antagonist (anti-blood sugar element) (before two contraction barium researchs). Other example comprises: CNS, Genitourinary (for example kidney, ureter, or bladder, vagina, uterus, fallopian tubal (for example contraceptive)), and biliary ductal tree (treatment of for example antiasthmatic, or gall-bladder fibre modification). Also have the example of other chamber to comprise surface (for example skin of local application and mucous membrane); Duct and middle ear (for example antibiotic, antivirotic), blood (for example intravenous injection, and transdermal and thoroughly muscle be delivery carrier). Other example of chamber comprises in the blood vessel, outside the blood vessel, and celiolymph, bronchial lumen, thoracic cavity, abdominal cavity, eyes zone, regional area, urethra, and genital tract, vagina for example, ureter, and fallopian tubal. Size

The size that multivalence is presented thing can affect action period as dissolubility (as mentioned above), therapeutic index, chamber, some character of removing effect aspect.

Term " size " comprises the molecule according to carbohydrate (being Stokes radius) and molecular weight of albumen (kD) size. The above molecular size of 60kD or present thing greater than the multivalence of the mean hydrodynamic diameter of 50 dusts and probably be easier to by chamber than micromolecular. In specific preferred embodiment, multivalence of the present invention is presented thing greater than 2kD and/or 5nM 50 dust mean hydrodynamic diameters.

According to " symptom " of wish treatment, the size of selecting multivalence to present thing can change. For example, use for parenteral route, low molecular weight compound (less than about 10,000MW) will be eliminated usually sooner. In addition, can use contain the cleavable attachment than large compound, this attachment connects enough little unit, when discharging calmly to be eliminated. Size can not be filtered by kidney effectively greater than the molecule of 60-70kDa albumen, and this is very important when multivalence is presented thing and is used in the blood flow. Oral when being used for, when lung or topical application, this material can not need to be eliminated or vivo degradation.

For zero medicine of removing basically, can not remove by kidney because they are too large, it is not absorbed by liver and removes, and when needs are removed, can bring out by many mechanism. For example, can by in serum with the attachment of remarkable speed hydrolysis in conjunction with low molecular weight fraction (the little molecule that its size is removed by kidney easily). In addition, can be by the natural attachment of material (for example enzyme) hydrolysis in the blood plasma that is present in conjunction with this low molecular weight fraction. In another example, attachment that can be by the medicine cracking is in conjunction with this low molecular weight fraction, and this medicine is removed multivalence and presented thing for (for example the second medicine of multivalence or unit price combination) need to take time. The example of second medicine can be mercaptan or chelating agent, can be disulfide bond and organic metal key to the connection of these medicine sensitivities.

Usually, high molecular weight material can not be by the oral blood that enters. The HMW system also is used for other purposes. In some instances, they can offer patient's (passing nose or lung mucous membrane after as the aerosol administration) by the transfer membrane infiltration. Some multivalence material can be ingested by the cell in the intestines, for example, and by the preparation of digestion process picked-up or with suppository dosing. Large multivalence medicine does not enter system's circulation and can be conducive to limit its side effect owing to it does not pass lung, intestines or respiratory tract. Additional embodiment

In certain embodiments, except with the unit price inhibitor, wish to present thing treatment patient with one or more multivalence. This unit price inhibitor may with or not with binding site B (be same as multivalence present group A is combined on the body) combination. For example, the unit price inhibitor of neuraminidase influenza (NA) (being present in the lip-deep hydrolase of influenza virus) raising polyacrylamide is presented the ability of HA inhibitor in case hemostasis cell agglutination (Choi, S.-K.; Mammen, M.; Whitesides, G.M. " chemistry and biology " (Chem.﹠ Bio.) 1996,3,97-104). NA site on virus surface is as second binding site of SA. Adding NA unit price inhibitor prevents the combination second time of SA, causes the effectiveness of these polymerization inhibitors to increase, and this may be owing to increased three-dimensional stability. In another other example, the present invention presents thing and can be used to be combined with other any methods for the treatment of.

The time that multivalence is presented the living effect of produce also is important. Multivalence hapten-carrier conjugate (it is not a part of the present invention in some instances) needs the long-time useful biological response that causes usually. Multivalence of the present invention is presented thing can be at 1 day, or in preferred 12 hours, in 5 hours, caused useful biological response or result for the treatment of in 1 hour, or need make an immediate response or tell in some cases, for example, need generation effect within second a few minutes to 1 during treatment asthma.

Further specify the present invention by the following example, it is not as restriction explanation of the present invention. All literature contents of in whole this application, quoting as proof, the manuscript that provides (referring to appendix A and B), autre action pendante patent application and publication (comprising " background " part) are clear and definite incorporated by reference at this. The multivalence that embodiment embodiment 1 is conducive to treat influenza is presented the preparation A part of thing-present as the hemagglutinative multivalence inhibitor of influenza-mediation generation and the on the spot evaluation in poly-(acrylic acid) derivative storehouse of saliva glycosides

Developed poly-(acrylic acid) that simple, a miniature preparation and evaluation present the mixture that affects its bioactive side chain (pAA) derivative storehouse. In this way, generation has N-acetylneuraminic acid (NeuAc-L-NH₂) use influenza virus A (X-31) to suppress the ability of the hemagglutination (HAI) of chicken red blood cell as pAA derivative-PAA (NeuAc-L) and the mensuration of side chain; Constant (the K of this inhibition is described^HAI _i) can calculate according to total NeuAc base in the solution. Use NeuAc-L-NH₂Different primary amine RNH with 26₂One of combination, generate and measure various ternarys-polymer pAA (NeuAc-L; R) (χ^NeuAc-L ～0.05；χ ^R～0.06). These polymer pAA (NeuAc-L; R) belong to the new polymerization multivalence saliva glycosides of a class, it is that influenza virus is absorbed into erythrocytic effective inhibitor.

The method with by with various amine RNH in water₂Reaction from poly-(acrylic anhydride) (pAAn) to the basis that is converted into of pAA derivative. Prepare the polymer that these are derived by direct ultrasonic reaction thing in the hole of little titer plate; Then need not further to process in the same plate, to measure.

Multivalence by virus protein hemagglutinin (HA) and the NeuAc part of expressing at the cell surface bunch surface that is absorbed into mammalian cell that interacts begins to infect influenza. Usually, monomer saliva glycosides be weak virus absorb inhibitor (suppressing to measure by hemagglutination) (referring to, Sauter for example, the people such as N.K., " biochemistry " (Biochemistry), 1989,28,8388).

In the little titer plate of 250 μ L (eq 1 is referring to appendix A) hole, process (sonicating) poly-(acrylic anhydride) (pAAn) suspension (0.12mg/ μ L) and amine RNH by sound wave₂The aqueous solution (0.1M) is synthetic to have a plurality of R groups as the pAA (R) of side chain. With different molar equivalents (mol eq.) NeuAc-L-NH₂Pass through NeuAc-L-NH with pAAn (eq.1)₂(1,2,3 or 4) and poly-(acrylic anhydride) (pAAn) reacts the solution for preparing copolymer pAA (NeuAc-L). The polymer of mol eq.=0 is to process (hydrolysis) separately at PBS buffer solution (137mM NaCl, 2.7mM KCl, 7.7mM Na from sound wave₂HPO ₄， 1.5mM KH ₂PO ₄，0.05％NaN ₃, pH 12) in the homopolymers pAA that obtains of pAAn. The copolymer pAA (NeuAc-L) of mol eq.＞0 is prepared as follows in little titer plate of 96 taper bottom outlets: (i) 6mg pAAn is placed in the hole; (ii) with variable (19-100 μ L) 0.1M NeuAc-L-NH₂PBS buffer solution (pH 12) soak powder; (iii) seal immediately plate and (use parafilm^Pat four sides of plate and cover tightly), then ultrasonic (the ultrasonic wish type of Fisher washer) mixture is 0.5 hour: ultrasonic the temperature of water in the bath (and reactant) is slowly risen to～50 ℃. By add among the 60 μ L 1.0M NaOH and the hole in the copolymer solution (pH～3) that produces to pH～7, before HAI measures, be adjusted to 100 or 200 μ L (cumulative volume) with PBS pH7. Above-mentioned method is extended to preparation ternary polymer pAA (NeuAc-L easily; R), mixture (pAAn (6mg), the 50 μ L 0.1M NeuAc-L-NH of 3 components of sound wave processing here₂(1 or 3) and 30 μ L 0.2M RNH₂)。

By measuring the freeze-drying reactant mixture¹H-NMR(D ₂O) spectrum and gel permeation chromatography characterize gained pAA (R) (Mw=39.5kDa, polydispersity=1.91; The polysaccharide standard). By comparing before ultrasonic and ultrasonic rear and free RNH₂The overall strength of background NMR signal, the RNH of the combination that observation produces₂ By peak (peak shape of polymer attachment is relatively wide) and chemical shift easily from unreacted RNH2's¹R's among the HNMR signal resolution pAA (R)¹The HNMR signal is (near the CH of acid amides group₂Be moved to low with the δ value of CH group). RNH as the combination of acid amides group₂Percentage with RNH₂Be the basis:With 5 kinds of different amine RNH₂(PABA, 6-aminocaprolc acid, the N-methyl hydroxylamine, (L)-arginine and 1 (NeuAc-L1-NH₂)) test the mean value that obtains and be～90 % (± 5). Because acid amides forms and the anhydride group hydrolysis is that competition occurs, the effectiveness of front a kind of method is subjected to various RNH₂Relative reactivity impact, and to RNH₂The pH of the aqueous solution (to fragrance and fatty amine, best pH difference～7 and 12) and RNH₂Than pAAn molar equivalent number (best mol eq.＜0.2) sensitivity.

The copolymerization derivative of pAA (pAA (1)-pAA (4)) is combined to method with semisolid, and the method is introduced NeuAc-L as side chain preparation (1-4 by the NeuAc derivative that use has different linking groups; NeuAc-Ln-NH₂ Referring to accompanying drawing 1 or flow process 1) (for the synthesis of NeuAc-L-

NH

₂1 and 2, referring to Sparks, M.A.; Williams, K.W.; Whitesides, G.M. " pharmaceutical chemistry magazine " (J.Med.Chem.) 1993,36,778; Ogura, H. waits people's " carbohydrate compound research " (Carbohydr.Res.) 1986,158,37; Lees, the people such as W.J. " JACS " (J.Am.Chem.Soc.) 1994,37,3419.NeuAc-L-NH₂(3,4), it is prepared as follows. 3 and 4 are used to be incorporated into the side chain of polymer, because can improve the binding affinity in monomer NeuAc-L and HA site: Watowish, S.J. at the aromatic portion of the centre that connects; Skehel, J.J.; Wiley, D.C.Structure 1994,2, and 719).

NeuAc R=OH; N-acetylneuraminic acid or sialic acid 1 R=(CH₂) ₃S(CH ₂) ₂NH ₂； NeuAc-L ₁-NH ₂ 2 R＝O(CH ₂) ₂O(CH ₂) ₂NH ₂； NeuAc-L ₂-NH ₂

Flow process 1

After sound wave is processed, use assay method according to chicken red blood cell and influenza virus A (X-31) (referring to for example Choi, S-K.; Mammen, M.; Whitesides, G.M. " chemistry is with biological " (Chem.﹠ Biol.) 1996,3,97, Lees, W.J. " pharmaceutical chemistry magazine " (J. Med.Chem.) 1995,37,3429; And being hereby incorporated by reference) hemagglutination of estimating immediately the polymer crude product solution suppresses (HAI) activity.

Following table 9 provides NeuAc-L-NH₂K when various molar equivalent^HAI _iValue (in the solution, preventing the least concentration of the NeuAc-L group of hemagglutinative pAA (NeuAc-L)). NeuAc-L-NH₂Molar equivalent directly with the mol ratio χ that in polymer, contains side chain Neu-Ac^NueAc-LRelevant. Table 9 shows that also the HAI of 3 kinds of other derivatives (pAA (2)-pAA (4)) in (sub) mM scope of pAA is active. Comparatively speaking, the HAI activity of all monomer sialic acids (1-4) is all low, K^HAI _i≥5mM。

Table 9 shows that the hemagglutination of pAA (NeuAc-L) suppresses active and pAA (NeuAc-L; R) storehouse.

The hemagglutination of table 9 pAA (NeuAc-L) suppresses active and pAA (NeuAc-L; R) storehouse

NeuAc-L-NH ₂Polymer RNH₂Molar equivalent molar equivalent K^HAI _i(μM) ² pAA(1) 0 15000 ^b

0.04(1) 27

0.06(1) 13

0.08(1) 3.9

0.10(1) 3.4

0.11(1) 4.4

0.12(1) 1.1

0.14(1) 1.1

0.17(1) 0.50

0.21(1) 0.30 pAA(2) 0.11(2) 0.80 pAA(3) 0.11(3) 0.20 pAA(4) 0.11(4) 3.1 pAA(1；R) 0.12(RNH ₂) 0.10(1)

RNH ₂

3-amino benzoic Acid 1.5

The amino 5-hydroxybenzoic acid 3.1 of 3-

PABA 3.1

4-amino-2-hydroxybenzoic acid 3.1

4-aminobenzenesulfonic acid 1.5

2-amino-nicotinic acid 1.5

N-methyl hydroxylamine 1.5

(D)-2-amino-1,5-anhydroglucitol 2.2

(D)-2-amino-2-deoxymannose 0.055

1-amino-1-cyclopropyl carboxylic acid 1.1

1-amino-1-encircles penta carboxylic acid 0.20

1-amino-1-cyclohexane carboxylic 0.028

Aminocyclohexane 0.0043

(L)-arginine 1.5

(L)-glutamate 2.5

Table 9 (continued)

(L)-histidine 1.5

(D)-4-hydroxy-proline 1.5

(DL)-leucine 0.30

(L)-phenylalanine 0.024

(L)-4 '-nitrobenzene alanine 0.048

(L)-phenyalanine methyl ester 0.024

1-amino-2-diphenylphosphino ethane 0.0021

(L)-3-(2 '-naphthalene) alanine 0.00050

(L)-tryptophan 0.0043

(L)-Gly-(L)-Gly-(L)-Gly 3.1

(L)-Gly-(L)-Phe 1.5 pAA(3；R) 0.13(RNH ₂)0.11(3)

RNH ₂

1-amino-2-diphenylphosphino ethane 0.0015

(L)-3 the multivalence of two types of " A " groups of (2 '-naphthalene) alanine 0.00070 B part-have is presented thing

Process NeuAc-L-NH by sound wave₂，RNH ₂With three kinds of compositions of pAAn A method (formula 1) is partly expanded to preparation ternary-polymer, pAA (NeuAc-L; R) (its multivalence is presented NeuAc-L and another R).

Table 9 (on) summarized from NeuAc-L-NH₂The RNH that (molar equivalent=0.10) is different from 26 kinds₂One of (molar equivalent=0.12) is in conjunction with obtaining pAA (NeuAc-L; R) K^HAI _iValue.

Several pAA (1; R) (and pAA (3; R)) specific activity that shows does not wherein have the parent copolymer pAA (1) of R group to improve about 7000 times of 100-, and (note: HAI measures to be needed to use certain virus, and can not Accurate Measurement K^HAI _iThe efficient of the inhibitor of value＜1nM). Although we are from crude product pAA (NeuAc-L; R) directly measure its activity, but several blank assays confirm that these ternary-polymer (not being pAA (NeuAc-L)) have high activity. General adding hydrophobic or the fragrant amido acid derivative improves activity greatly. Our verified some architectural feature (not only hydrophobicity) is for reducing K^HAI _iBe worth effective especially. We sum up some non--saliva glycosides group, although itself do not show that HAI is active, it improves pAA (NeuAc-L; R) activity is up to about 10⁴Doubly.

Best pAA (NeuAc-L; R) belong to the new hemagglutination inhibitor of a class, it has high activity usually when the middle equimolar ratio of relative NeuAc-L (about 5%) and R (about 6%): the NeuAc-L of per 1% mol ratio or R equal about 6 side chains (every polymer molecule). The importance in conjunction with side chain is emphasized in regulating these multivalence and present the activity of thing in this discovery.

The method of describing in A B part flow process is synthesized in by the hole at little titer plate and is measured the pAA derivative and estimate the biologically active of these polymer. These methods are so that exist many polymer storehouse screenings when controlling mol ratio in conjunction with side chain easy. Because little titer plate is determined at biological and pharmaceutical science in be conventional, the aggegation that the method is generally used for screening and obtain to be subjected to the multivalence inhibitor and affects interacts and guide's material (leads) of other method. This method is synthetic and screens the method for the quick economy of bioactive multivalence inhibitor. Embodiment 2-is conducive to suppress the preparation that the A multivalence of platelet aggregation is presented thing

RGD (L-Arg-L-Gly-L-Asp) is the recognition sequence that fibrinogen is combined with blood platelet. The RGD analog suppresses the interaction between fibrinogen and the platelet membrane glycoprotein iib/iiia compound, therefore as the platelet aggregation inhibitor (referring to Pillips, D.R. and " cell " (Cell), 65:359 (1991)).

Existing RGD to present thing as the multivalence of part is prepared as follows: will be fixed on RGD on the solid carrier of C-end according to synthetic (SPPS) method preparation of known solid-phase peptide, for example on the Wang resin. Be loaded with the RGD of solid and the RGD of the succinylation that the succinyl oxide reaction is fixed, then with 1 of list-BOC-protection, 6-diamino hexane (for example DCC/HOBt) reaction. The RGD that derives by acid treatment cracking from the resin and the accessory substance (concomitant) of removing BOC, then purifying (for example passing through HPLC) obtains the RGD with the coupling part functionalization with terminal amino group. Polysuccinimide (is prepared according to known method: referring to for example United States Patent (USP) 5,484,878 prepare polysuccinimide from lucid asparagus) be suspended in the solvent such as dimethyl formamide (DMF) or dimethylacetylamide, the RGD of bridging agent-functionalization is added in the suspension, stirs simultaneously. After the amino solution of polysuccinimide was finished, with the not amino polysuccinimide of separating of hydrolysis, impurity was removed in dialysis with the aqueous alkali diluted reaction mixture, obtains having poly-(asparatate) main chain that carries RGD bridging agent functionalization. The controls such as the amount of the pH by the conditioned reaction mixture, the RGD-bridging agent that exists in reactant mixture, reaction time have the replacement degree of the main chain of RGD, the size of poly-(asparatate) polymer that produces by the amino solution.

Poly-(asparatate) multivalence of screening gained RGD-is presented thing, and according to means known in the art, for example the platelet aggregation determination method is measured multivalence and presented thing to the impact of fibrinogen polymerization platelet membrane glycoprotein iib/iiia compound. Embodiment 3-prevents the abbreviation of the erythrocytic multivalence polymerization of ricin adhesion galactoside

The PBS-phosphate buffered saline (PBS); The Et-ethyl; MeOH-methyl alcohol; H-hour; The i-Pr-isopropyl; PBMA-butadiene maleic acid copolymer; The RCA-ricinus agglutinin; The hemagglutinin of BHA-bromelain enzymatic lysis; The LDH-lactate dehydrogenase. Conventional method

All general chemistry product that use obtain not through being further purified, unless note is arranged in addition from supplier. Except as otherwise noted, all chemicals are from Aldrich Chemical Co St. Louis, and Missoure buys. 2 the week age chicken red blood cell (RBCs or red blood cell) buy from Spafas Inc.. With red blood cell (suspension (about 5%v/v) form with store buffer liquid provides) phosphate buffered saline (PBS) (PBS:137mM NaCl, 2.68mM KCl, 7.75 mM Na₂HPO ₄，1.47mM KH ₂PO ₄, pH 7.2) washing then be suspended among the PBS (about 5 %v/v). Ricin (the RCA of FITC-mark that comprises fluorescein isothiocyanate (FITC)-mark₁₂₀, the RCA of FITC-mark₆₀) ricin (RCA₁₂₀， RCA ₆₀) buy from Sigma Co..

Gal-βO-L ₁NH ₂With Gal-α C-L₂NH ₂(flow process 1) synthetic: in carrene (180mL) solution that contains β-D-galactolipin pentaacetic acid ester (7.8g, 19.98mmol) and allyl alcohol (5.6ml, 58.82mmol) in the cooling ice bath, drip BF₃·Et ₂O (4.0mL, 32.32mmol). Stir (4h, 0 ℃; Then 30h, about 20 ℃) after, mixture is poured into the saturated NaHCO of cooling₃(200mL). After the vibration, tell organic phase, and use again saturated NaHCO₃(200mL) washing. Use MgSO₄After the drying, vacuum steams dichloromethane solution and produces the light yellow oil (5%MeOH/CH of Rf=0.57₂Cl ₂) crude product is dissolved in the methyl alcohol (100mL), then add the water (50mL) of LiOH (2.88g, 120mmol). Stir (12h, about 20 ℃), by adding 6.0MHCl (about 20mL) neutralization reaction mixture. The aqueous solution of vacuum concentrated mixture obtains the viscous crude residue, and it is used flash column chromatography purifying (silica gel; 5% to 40%MeOH/CH₂Cl ₂). Obtain product Gal-β OCH₂CH＝CH ₂, two step gross production rates are 91% (4.0g), are yellow oil. (Rf=0.80 in 30% MeOH/CH₂Cl ₂)。 ¹H-NMR(300.1MHz，CD ₃OD)：δ(ppm)6.00- 5.89(m，1H)，5.36-5.29(dd，J＝17.2，3.2，1H)，5.18-5.12(dd，J＝12.1， 3.2Hz，1H)，4.40-4.34(dd，J＝13.0，5.2Hz，1H)，4.27-4.25(d，J＝7.3Hz， 1H；H _lax), 4.17-4.10 (dd, J=13.0,6.1Hz, 1H), 3.90-3.83 (m, 1H), 3.80-3.68 (m, 3H), 3.56-3.50 (m, 2H), FAB-MS (glycerine): m/z 221[M+H]⁺ HRMS: calculated value C₉H ₁₇O ₆221.1024 measured value 221.1025.

With Gal-β OCH₂CH＝CH ₂(4.0g，18.17mmol)，HSCH ₂CH ₂NH ₂HCl (6.19g, 54.5mmol) and 4, then the water (50mL) of 4 '-nitrine two (4-cyanopentanoic acid) (0.4g, 1.43mmol)-methyl alcohol (5mL) solution vacuum outgas 10 minutes use N₂Saturated (with N₂Blasted solution 30 minutes). The reaction flask that will contain mixture is placed on (Rayonet ) in the photochemical reactor, then 254nm irradiation 10 hours. Shine mixture by adding 2.0M NaOH (28mL) neutralization, and volatile matter is removed in evaporation immediately. Evaporation obtains light yellow oil, and it is used flash column chromatography purifying (10%MeOH/CH₂Cl ₂To 5% i-PrNH₂/40％MeOH/CH ₂Cl ₂). Obtain oily adduct (Gal-β O-L₁NH ₂)。 (Rf＝0.33 in 5％I-PrNH ₂30％ MeOH/CH ₂Cl ₂.). ¹H-NMR(250.1MHz， CD ₃OD)：δ(ppm)4.22-4.19(d，J＝7.4Hz，1H；H _lax)，4.0-3.95(ddd，J＝9.9， 6.1，6.1Hz，1H)，3.87-3.81(m，1H)，3.78-3.63(m，4H)，3.53-3.43(m，2H)， 2.81-2.76(t，j＝6.5Hz，1H)，2.67-2.59(q，J＝7.1Hz，4H)，1.92-1.82(quin， J＝6.6Hz，2H)； ¹³C-NMR(100.6MHz，CD ₃OD):? (ppm) 76.6,75.8,72.5,70.3,69.2,62.5,41.6,35.6,31.2 29.0; FAB-MS (glycerine): m/z 298 [M+H]⁺ HRMS: calculated value C₁₁H ₂₄NO ₆S 298.1323, measured value 298.1324.

Gal-αC-L ₂NH ₂：Gal-αCCH ₂CH＝CH ₂The α of preparation by beta galactose five nicotinates-C-allylation, then (Giannis, A. are carried out in the hydrolysis of acetic acid esters; Sandhoff, K. " tetrahedron communication " (Tetrahedran Lett.) 1985,26,1479-1482). (¹H- NMR(400.0MHz，CD ₃OD)：δ(ppm)59.1-5.83(m，1H)，5.13-4.99(m，2H)， 4.00-3.96(m，2H)，3.95-3.88(m，1H)，3.76-3.67(m，4H)，2.49-2.34(m， 2H)； ¹³C-NMR(100.6MHz，CD ₃OD): δ (ppm) 136.7,116.9,75.7,74.0,71.9,70.1,70.0,62.0,31.0; FAB-MS (glycerine): m/z 227[M+H]⁺ From Gal-α CCH₂CH＝CH ₂Prepare as mentioned above Gal-α C-L₂NH ₂。 ¹H-NMR(400.0MHz， CD ₃OD)：δ(ppm)3.95-3.63(m，7H)，3.12(t，J＝6.9，2H)，2.81(t，J＝3.4， Hz，2H)，2.66-2.59(m，2H)，1.81-1.62(m，4H)， ¹³C-NMR(100.6MHz， CD ₃OD): δ (ppm) 75.7,74.0,71.9,70.4,70.2,62.5,39.9,32.1,26.8,24.8,20.9; FAB-MS (glycerine): m/z 282[M+H]⁺ HRMS: calculated value C₁₁H ₂₄NO ₅S 282.1375 measured values 282.1374.

PAA (Gal-β), pBMA (Gal-β), and pBMA's (Gal-α) (accompanying drawing 5) is synthetic: pAA (Gal-β): following synthetic pAA (Gal-β; 0.4) method be the conventional method of synthetic pAA (Gal). Poly-to containing (N-propionyloxy succinimide) or pNAS (500mg equals 3mmol NAS) (Mannen, M.; Dahmann, G.; Whitesides, G.M. " (pharmaceutical chemistry magazine " (J.Med.Chem.) 1995.38,4179) DMF (DMF, 8mL) solution in add the Gal-β O-L that is dissolved among the DMF (2mL)₁NH ₂(356mg, 1.2mmol) then adds i-Pr₂NEt (0.2mL, 1.2mmol). After stirring (2h, about 20 ℃), add 1.0M NaOH (3mL) alkalization mixture, then stirred again 2 hours at about 20 ℃. Reaction is transferred to (the about 12-14kDa of MW cutoff in the bag filter with reactant mixture after finishing; Spectrum  derives from Sprcturm Medical Industries, Inc.), and at about 20 ℃ of dialysis 3 hours: H₂O (2 * 4L), 0.05M NaOH (4L), 0.5M NH4Cl (4L), and H₂O (2 * 4L). Material in the freeze-drying bag obtains pAA (Gal-β; 0.4), it is fluffy white solid (499mg).¹H-NMR(500.1MHz，D ₂O)：δ(ppm)4.3(d， J＝7.5Hz，H _lax), 3.89 (br s), 3.85 (s), 3.8-3.5 (m), 3.4 (br m), 3.3 (br s), 2.6 (br s), 2.2-1.9 (br d), 1.7-1.3 (br m); %S: calculated value pAA (Gal-β; 0.4) 6.97, measured value 282.1374. Do you prepare pAA (Gal-according to following identical method?). %S:pAA (Gal-β; 0.2) calculated value 5.00, measured value 5.12; PAA (Gal-β; 0.6) calculated value 8.02, measured value 8.06; PAA (Gal-β; 0.8) calculated value 8.67, measured value 8.58; PAA (Gal-β; 1.0) calculated value 9.11, measured value 9.05.

PBMA (Gal-β), and pBMA's (Gal-α) is synthetic: according to synthetic these polymer of above-mentioned slightly diverse ways, wherein substitute poly-(butadiene-altogether-maleic acid) or pBMAn with pNAS as precursor polymer. PBMAn (solution with acetone provides (Polysciences, the Inc.)) vacuum drying of five equilibrium also is dissolved in DMF before use again. PBMA (Gbl-β; 0.09):¹H-NMR(500.1MHz，D ₂O)：δ(ppm)5.6(br s)， 5.4(br s)，3.9-3.5(br m)，3.4-3.3(br m)，3.0-2.9(br m)，2.7-2.5(br m)， 2.4-2.1(br s)，1.8(br s，3.9-3.5(br m).pBMA(Gbl-α；0.09)： ¹H- NMR(500.1MHz，D ₂O): δ (ppm) 5.6 (br s), (5.5-5.3 br d), (4.3-3.7 br m), (3.6 br m), 3.3 (br s), 2.9 (br s), (2.8-2.4 br s), (2.4-2.0 br s), 1.8-1.4 (br m), 1.0 (br s). %S:pAA (Gal-α; 0.05) calculated value 1.62, measured value 1.55; PAA (Gal-α; 0.09) calculated value 2.62, measured value 2.69; PAA (Gal-α; 0.17) calculated value 4.11, measured value 4.12; PAA (Gal-α; 0.22) calculated value 4.81, measured value 4.85.

The agglutination of the ricin mediation of chicken RBCs, and prevent by pAA (Gal): (i) ricin and chicken RBCs adhesion: with RBCs (0.5%v/v, 0.4mL) PBS, the RCA of pH7.2 and fluorescence ricin FITC-mark in 1mL Eppendorf bottle₁₂₀(40nM) or the RCA of FITC-mark₆₀The PBS solution of (1.4 μ M) fully mixes. At 4 ℃ of incubations after 2 hours, with mixture at 2000rpm centrifugal 2 minutes, wash red blood platelet with 1.0mL PBS, and be suspended in gently again among the PBF (0.2mL). Be placed on the absorption of the RBCs that obtains the RCA absorption on the sheet glass and the light figure of fluorescence by the suspension blood platelet with five equilibrium, with optics and fluorescence microscope (Leica DMRX) test sample.

(ii) cell that makes RBSc avoid ricin with pAA (Gal-β) absorbs: with the RCA of FITC-mark₁₂₀(80nM; 0.2mL) PBS solution and pAA (Gal-β in the Eppendorf bottle; 0.4) (90gmL^-1Or [Gal] equals about 200 μ M) PBS solution mix. Behind the incubation (30 minutes, 4 ℃), ricin-polymeric blends is added to RBSc (0.5%v/v; 0.4ml) PBS suspension in, then stir gently and 4 ℃ of incubations 2 hours. With the mixture of incubation at 2000rpm centrifugal 2 minutes. Wash with 1.0mL PBS removing the red blood platelet that obtains after the supernatant, and be resuspended among the 0.2mL PBS, then detect with light microscope.

Hemoagglutination (ricin-bring out) suppresses (HAI) and measures: with the serial dilution liquid of 2 times of 50 μ L ricin solution (mg mL-1) at 12 holes (little titer plate at the bottom of the taper of 8 * 12-hole circle; ICN Flow) measures the PBS solution (RCA of the ricin of preparation in₁₂₀＝16nM；RCA ₆₀=1.9 μ M) titer. In each hole, add 50 μ LPBS, then add PBS (the 100 μ L) solution of chicken red blood cell. Mixed solution and about 20 ℃ of incubations 1 hour. The terminal point of hemagglutinin (HA) is defined as metapore, and the ricin aggegation red blood cell of q.s is wherein arranged. With polymerization galactoside (1-2mg mL^-1 [Gal] about 2-6mM) or the deposit PBS solution of unit price galactoside (5mM) (50 μ L) decide hole 2-times serial dilution by 12 droplets that contain 50 μ LPBS. Behind the galactoside solution serial dilution of polymerization or monomer, each hole (50 μ L) and 50 μ L RCA₁₂₀(16nM) or RCA₆₀(1.9 μ M) mixes. , after 30 minutes 100 μ L chicken red blood cell suspension (0.5v/v) are added in each hole at about 20 ℃ of incubations, then stir gently and incubation (1 hour, about 20 ℃). The terminal point of HAI is last hole, wherein observes the blood platelet of aggegation. This terminal point (K^HAI _i) be defined as the Cmin of the galactoside in the solution that suppresses the RCA that ricin brings out. According at least 15 independent experiment calculating Ks^HAI _iValue. Results and discussions

The synthetic polymer that has galactoside: synthesize two derivatives (the Gal-β O-L as the D-galactoside of polymerization multivalence D-galactoside monomer precursor₁NH ₂With Gal-α C-L₂NH ₂)：Gal-βO-L ₁NH ₂Containing β between galactoside (Gal) group and amine-terminal connector (flow process 1)-O-connects; Gal-α C-L₂NH ₂Contain α-C-glucoside. Select Gal-α C-L₂NH ₂As the C1-epimer analog of β-O-galactoside, be because it can be easily with highly-solid selectively and on a large scale preparation. Gal-α C-L₂NH ₂The C-glucoside connect the additional advantage that antagonism chemistry and enzymolysis are provided. With two epimers of D-galactoside binding affinity of itself and ricin relatively. Two types polymer-poly-(acrylic acid) (pAA: Mw=～140kDa, Mw/Mn=1.91) (Mammen, M.; Dahmann, G.; Whitesides, G.M. " pharmaceutical chemistry magazine " (J.Med.Chem.) 1995,38,4179-4190) and butadiene maleic acid copolymer (pBMA; Mw=10-15kDa) be used as the polymerization support to present the multiple replicated architecture as the unit price galactoside of side chain. These polymer are called pAA (Gal-β), pAA (Gal-α), pBMA (Gal-β), and pBMA (Gal-α). Select the pAA-based polyalcohol to estimate that it is relatively scalable when high ionic strength at least; The polymer that expectation pBMA-derives is not too variable. Pass through poly-(N-propionyloxy succinimide) (pNAS) (Mammen, M. with above-mentioned method; Dahmann, G.; Whitesides, G.M. " pharmaceutical chemistry magazine " (J. Med.Chem.) 1995,38 is 4179-4190) with Gal-β O-L₁NH ₂Reaction in DMF (about 20 ℃ 2d), and make to react with excessive 1.0M NaOH and stop to obtain pAA (Gal-β). By pNAS and the χ equivalent Gal-β O-L that makes every equivalent active ester groups on the polymer₁NH ₂Reaction (χ=0.2,0.4,0.6,0.8 and 1.0) prepares various pAA ' s (Gal-β; 0.2-1.0). Parameter χ also equals the mol ratio of Gal in the polymer, and is defined as containing the quantity of Gal side chain divided by side chain sum (flow process 1). Identical strategy is used to pBMA (the Gal-β that butadiene maleic anhydride copolymer (pBMAn) produces; 0.05-0.22) and pBMA (Gal-α; 0.05-0.22). (MW blocks～3.5kDa) purifying, and utilization all polymer by dialysis¹H-NMR spectrum and combustion analysis (sulphur) characterize. According to the combustion analysis of polymer, the productive rate that the acid amides of Pnas and pBMAn forms reaction is respectively 〉=95% (for pNAS) and 〉=65%.

The polymer of presenting galactoside suppresses ricin and erythrocytic combination: with 2 age in week chicken red blood cell (RBCs) be used as the model system of mammalian cell. Red blood cell is seedless, and synthetic proteins not. But they provide the good model of ricin targeted cells, and the system of research ricin and cell surface adhesion is provided: there are many sugared conjugate (the about 2-3 of everyone RBC * 10 that contain the beta galactose glycosides in erythrocyte surface⁶The Gal residue) (Sandvig, K.; Olsnes, S.; Phil, " biochemical society's will " (A.J.Biol. Chem.) 1976,251,3977-3984).

Obtaining representative absorption and fluorescent graphic and proof ricin is combined with RBCs and causes its aggegation and dissolving. Confirm that with the ricin of fluorescence isothiocyanate (FITC) mark aggegation and dissolving are because the effect of ricin. Also prove multivalence galactoside pAA (Gal-β; 0.4) stop these effects.

The little chicken red blood cell of (PBS) pH7.2 (0.5% volume ratio) and ricin (RCA with the phosphate buffered saline (PBS) form of suspension₁₂₀～16nM；RCA ₆₀～1.9 μ M), measure the agglutination that the polymerization galactoside suppresses the ricin mediation. Table 10 has been summarized the carbohydrate containing side chain (χ of various mol ratios^Carbohydrate) the hemagglutination of polymer of purifying suppress (HAI) active K^HAI _i(being defined as the least concentration that stops the required inhibitor of hemagglutination).^aK ^HAI _iValue refers to suppress the least concentration of polymer that ricin brings out the carbohydrate containing side chain of chicken RCA. Each value represents the independent mean value of measuring 5 times; The experiment uncertainty of each value is about ± and 50%.^bShown in concentration do not observe inhibition.^cThis value representative can suppress the solution concentration that contains polymer that ricin brings out the carboxylic acid side chain of chicken RCA.

Table 10 multivalence polymerization galactoside brings out the chicken RCA to ricin inhibitory action

Inhibitor	K _i ^HAI(μM) ^a		Inhibitor	K _i ^HAI(μM) ^a
	K _i ^HAI(μM) ^a			K _i ^HAI(μM) ^a		RCA ₁₂₀	RCA ₆₀	RCA ₁₂₀	RCA ₆₀
	Gal-β-Ome Gal-α-Ome Gal-βO-L ₁NH ₂ Gal-αC-L ₂NH ₂ GlcNAc-βO-L ₁NH ₂ PAA(Gal-β；0) PAA(Gal-β；0.2) PAA(Gal-β；0.4) PAA(Gal-β；0.6) PAA(Gal-β；0.8) PAA(Gal-β；1.0)	200 400 37 250 ＞9000 ^b ＞35000 ^b，c 0.16 0.14 0.18 0.27 0.56		42 50 16 39 ＞9000 ^b ＞35000 ^b，c 8.1 17 20 42 180	pBMA(Gal-β；0) pBMA(Gal-β；0.05) pBMA(Gal-β；0.09) pBMA(Gal-β；0.17) pBMA(Gal-β；0.22) pBMA(Gal-α；0.05) pBMA(Gal-α；0.09) pBMA(Gal-α；0.17) pBMA(Gal-α；0.22) pAA(GlcNAc-β；0.2) pAA(NeuAc-α；0.2) pBMA(NeuAc-α；0.2)	RCA ₁₂₀	RCA ₆₀	RCA ₁₂₀	RCA ₆₀	＞30000 ^b，c 2.0 14 5.0 0.73 ＞160 280 61 12 ＞290 ^b ＞170 ^b ＞200 ^b	≥30000 ^b，c 1.0 12 ＞80 ≥94 ≥160 33 17 5.0 ＞290 ^b ＞170 ^b ＞200 ^b

Unit price Gal-β O-L₁NH ₂The anti-RCA that provides₁₂₀And RCA₆₀K^HAI _iValue is low 5 times and 3 times than Gal-β-OMe respectively. Contain the good of beta galactose glycosides corresponding to the specific activity of unit price galactoside of β-O-end group isomery configuration, although difference is little. Observed result shows, the combination of Gal residue and ricin Gal binding site to end group isomery configuration (β 〉=α) or to the atomic nature that adheres to end group isomery carbon atom (sensitiveness of O 〉=C) is not high.

PAA (Gal-β; 0.2-1.0) show anti-RCA₁₂₀The HAI of the aggegation of bringing out in solution with the Gal part that is lower than micro-molar concentration is active (by K^HAI _iValue representation). The anti-RCA of same polymer₆₀The anti-RCA of HAI specific activity₁₂₀Low about 50-300 doubly; Anti-RCA₆₀Active at most than 50 times of monomer Gal derivative. By contrast, pAA (Gal-β; 0.4) anti-RCA₁₂₀Specific activity unit price Gal-β-OMe's is high approximately 1500 times, than Gal-β O-L₁NH ₂High approximately 270 times. PBMA (the Gal-β of (Asia) micro-molar concentration; 0.05-0.22) show anti-RCA₁₂₀And RCA₆₀HAI is active; Anti-RCA₁₂₀Activity be better than anti-RCA₆₀. But, pBMA (Gal-α; 0.05-0.22) anti-RCA₆₀The anti-RCA of HAI specific activity₁₂₀Make an appointment 4 times. In the multivalence galactoside that pBMA makes up, pBMA (Gal-β; 0.22) and pBMA (Gal-α; 0.22) be respectively anti-RCA₁₂₀And RCA₆₀The strongest active inhibitor.

PBMA (Gal) is the polymer of less, and molecular weight is about 10-15kD, and a biocompatible polymer backbone (pBMA) (Conroy, C.W. are arranged; Wynns, G.C.; Maren, T.H., " Bioorganic Chemistry (Bioorg.Chem.) ", 1996,24,262-272).

Thing has also carried out presenting a plurality of identical non-galactoside carbohydrate (pAA (NeuAc-α to other polymer in contrast; 0.2) (Mammen, M.; Dahmann, G.; Whitesides, G.M., " medical chemistry magazine (J.Med.Chem.) ", 1995,38,4179-90), pAA (NeuAc-α; 0.2-1.0), pAA (NeuAc-β; 0.2-1.0)) test, but as a result neither one at table 10 specific concentration (K^HAI _i＞300 μ M) RCA that ricin is caused shows inhibitory action. Therefore reach a conclusion: the polymer that the RCA that ricin causes is presented galactoside suppresses selectively. The usefulness of α and β anomer seemingly equates. This presentation of results, being aggregated the galactoside inhibition mainly is because the single-minded combination of Gal acceptor site of galactoside part and ricin. We think that it is because the multivalence of galactoside part and a plurality of acceptor sites (entropy improves) combination that the multivalence galactoside of polymerization divides with respect to the active higher part of unit price galactoside.

Fig. 6 has gathered the HAI activity of polymer anti-ricin. Fig. 6 a is (about RCA₁₂₀) show the K of polymerization galactoside^HAI _iValue is the Gal (χ of this polymer^Gal) nonlinear function of mol ratio. At RCA₆₀The activity and the χ that suppress polymer in the hemagglutinative function^GalBetween relation as and if described RCA₁₂₀Inhibitory action is closely related. The K of polymer^HAI _i -χ ^GalCorrelation also depends on the type of polymer backbone: the curve smoothing of large and pliable and tough pAA (Gal-β); The curve of the pBMA (Gal) of little and expansion is sharp-pointed and concern for the part pseudo-linear. Present pAA (the Gal-β of low-density Gal side chain (20 Gal are arranged on each polymer chain approximately); 0.01) (degree of polymerization=DP about 2000) active K of showing^HAI _iBe about 2.0 μ M, than unit price Gal-β O-L₁NH ₂High 19 times. We are pAA (Gal-β; 0.01) with respect to unit price Gal-β O-L₁NH ₂The flexibility of its polymer backbone is given the credit in active increase, therefore, gives the credit to the ability that it regulates distance between the galactoside part; This flexible its multivalence with the ricin acceptor site that promotes is combined.

Be used for measuring RCA under the condition of ability that the polymer of presenting Gal suppresses agglutination at us₁₂₀Titre---can aggegation RBCs (200 μ L, 0.25%v/v is suspended in PBS solution) Cmin---be 4nM, RCA₆₀Titre be 480nM. Both difference shows RCA₆₀(B chain; About 3 Gal acceptor sites) cohesiveness compares RCA₁₂₀(two B chains; About 6 Gal acceptor sites) a little less than. Preventing the anti-RCA of unit price galactoside aspect the RCA that ricin is induced₆₀Than anti-RCA₁₂₀More effective. Owing to the inhibition of agglutination has been reflected the competitive binding of the Gal acceptor site of unit price galactoside and ricin, so the Discrepancy Description on this activity is for rare two hypothesis of being attached to of ricin and cell. The first, the anti-RCA of monomer galactoside that observes₆₀With anti-RCA₁₂₀The difference of HAI activity may be since the monomer galactoside to RCA₆₀And RCA₁₂₀The difference of the interior affinity of upper Gal acceptor site. But, with equilibrium dialysis and fluorescent technique several monomer derived things of galactoside and the result of study in conjunction with situation of two ricins are shown, two kinds of ricins have similar affinity (multiple that differs is less than 10) (Baenziger to the unit price galactoside, J.U., Fiete, D., " journal of biological chemistry " (J.Biol.Chem.), 1979,254,9795-9; Houston, L.L., Dooley, T.P., " journal of biological chemistry ", 1982,257,4147-51; Rivera-Sagredo, A., Solis, D., Diaz-Maurino, T., Jimenez-Barbero, J., Martin-Lomas, M., " european journal of biological chemistry " (Eur.J.Biochem.), 1991,197,217-228). The second, the anti-RCA of unit price galactoside₆₀K^HAI _iValue is than anti-RCA₁₂₀Low may be with suppressing aggegation required ricin egg from the number (n) of the Gal acceptor site that is blocked with respect to ricin on the sum of Gal acceptor site, i.e. n/3 (RCA₆₀) or n/6 (RCA₁₂₀) relevant. That is to say, if blocking-up RCA₆₀And RCA₁₂₀The Gal site of upper similar number, then RCA₆₀On the Gal site that is blocked account for larger proportion, therefore eliminating RCA₆₀The agglutinability aspect compare RCA₁₂₀More effective.

The K that the galactoside of polymerization provides^HAI _iValue is RCA₁₂₀Ratio RCA₆₀Low. Unit price is presented relative raising (that is, the K of polymer of the activity of middle galactoside part^HAI _iThe relative value of value and monomer) be RCA₁₂₀Ratio RCA₆₀Height. These presentation of results, multivalent ligand as inhibitor at anti-high chemical valence target (RCA₁₂₀) time than anti-low valent target (RCA₆₀) more effective.

For RCA₆₀, some polymer (pAAGal-β; 0.6-1.0), pBMA (Gal-β; 0.17-0.22), pBMA (Gal-α; 0.05)) aspect the aggegation of blocking-up ricin mediation (on each Gal base) than corresponding multivalence galactoside weak effect. These results support such conclusion, namely for the multivalence inhibitor of some polymerizations, and in conjunction with by BHA (3 acceptor sites), LDH (4 acceptor sites) and RCA₆₀The entropy of a small amount of acceptor site that (about 3 acceptor sites) present improves not obvious, and the part of presenting on the polymer backbone to the affinity of acceptor may be lower than the part that does not tie down polymer (may because the uncertain interaction between polymer backbone and the protein).

Many polymer than more effective this statement of facts of corresponding unit price galactoside, are to be determined K except acceptor site by the many factors partly capturing aspect the aggegation (on each Gal base) of blocking-up ricin mediation^HAI _iValue. Because each polymer is presented a plurality of Gal parts, so the combination of polymer and ricin certainly can be quite tight, even relative with the combination of each single Gal group weak. The situation that the Gal acceptor is occupied on the ricin is not directly measured in the experiment of presenting here, therefore, inhibition to hemagglutinative function both can be because these in conjunction with the combination that the entropy of the acceptor site of Gal improves, also can be because the effect that non-acceptor causes suppresses as leaning on the space.

The baroque glycoprotein that some is natural and oligosaccharides tightly are combined on the B chain of ricin. The glycopeptide of deriving from myosin (class is presented the glycoprotein of multiple kinds of carbohydrate bunch) by the enzyme effect is exactly an example: these compounds are to the ricin dissociation constant K of (with to the B chain)_dBe about 10 to 0.1 μ M (Baenziger, J.U., Fiete, D., " journal of biological chemistry " (J.Biol.Chem.), 1979,254,9795-9). These glycopeptides are multiple-limb oligosaccharides and oligopeptides of presenting a plurality of identical galactosides and GalNAc part. Therefore, according to distributing in the quantity of galactoside in each molecule and the molecule, containing galactoside sugar conjugate is multivalence on the structure, and we think that these structural properties can be explained the reason that higher binding affinity is arranged with respect to the polysaccharide galactoside for these glycopeptides of ricin. The present invention presents the polymer pAA (Gal) of galactoside and pBMA (Gal) has good composite character and hemagglutination suppresses active, and K^HAI _iValue is up to the multivalence material of about 0.1 μ M. These polymer are easy to synthesize, and chemistry and the enzymatic property of comparing these polymer of presenting the C-galactoside with the glycopeptide to the enzyme sensitivity that myosin is derived are metastable.

In a word, above-mentioned presentation of results, the synthetic polymer of presenting a plurality of identical β as side chain-D-galactolipin simple derivatives has suppressed the adhesion of ricin and chicken red blood cell effectively, and this result conforms to the quantitative measurment result that testing result and the hemagglutination of optics/fluorescence microscope suppress (HAI) determination method. In case with the unit price galactoside---their ricin-cytoadherence effects own weak inhibitor---change the multivalence galactoside of polymerization into, the galactoside residue of polymer combination is to RCA₁₂₀The HAI activity will increase about 10²To 10³Times, but to RCA ₆₀50 times of only increases, take the total concentration of these sugar units in measuring solution as benchmark. The activity of the galactoside of polymerization is subjected to various factors, contains the mol ratio of Gal side chain such as polymer, the C-1 epimerism configuration of galactoside (β 〉=α) and polymer backbone. The multivalence N-acetyl glucosamine of embodiment 4 polymerizations is induced

Mouse perforatorium exocytosis abbreviation: Gal-reverse transcriptase, β-Isosorbide-5-Nitrae-galactosyl reverse transcriptase; ZP, oolemma; GlcNAc, the N-acetyl glucosamine; Gal, galactolipin; PA, poly-(acrylamide); PAA, poly-(acrylic acid); PAA (GlcNAc-β) presents in the GlcNAc that a plurality of identical β as the acid amides side chain-O-connects poly-(acrylic acid); PAA (Gal-β) presents in the Gal that a plurality of identical β as the acid amides side chain-O-connects poly-(acrylic acid). Materials and methods

The multivalence N-acetyl glucosamine (GlcNAc) of polymerization and galactoside (Gal): present a plurality of identical as the GlcNAc of acid amides side chain or poly-(acrylic acid) (pAA (GlcNAc of Gal according to the similar method preparation of the multivalent compounds of above-described embodiment 3 preparation polymerizations; χ) and pAA (Gal; χ), wherein, χ=0.2,0.4,0.6,0.8,1.0). PAA (GlcNAc) refers to respectively present a plurality of identical GlcNAc-β-L as the acid amides side chain with pAA (Gal)₁NH ₂With Gal-β-L₁NH ₂Poly-(acrylic acid).

Mouse sperm acrosome reaction process:

The mouse sperm is adapted to 30min at 35 ℃, then be with or without in the presence of the polymer 37 ℃ of insulations. Each experiment has following control group:

1) only is used for estimating the phosphate buffer (between test period, be generally less than 5%, and will other deduct measuring from all) of " spontaneous " acrosome reaction degree;

2) the used quality of polymer backbone---poly-(acrylamide) or pA---is identical in quality with the GlcNAc's that is used for the required mole of generation pAA (GlcNAc);

3) Zona Pellucida Glycoprotein (ZP), " natural " inducer; And

4) at 0,15min, the several time points of 30min and 60min start the calcium ion carrier A 23187 of reactant mixture;

The sperm of acrosome reaction and ZP and ionophoric product (%) are respectively 27.1 (± 0.10) and 53.7. Poly-(acrylamide) or pA are used as negative control group and test: the concentration of the value representation pA formamide of X-axis. Sample of sperm is dry on sheet glass, to harmless acrosome dyeing, measures the ratio of staining cell and undyed quantity in 200 sperms of each sample. Result and discussion

The acrosome exocytosis that multivalence GlcNAc induces:

Fig. 8 gathered unit price GlcNAc in inducing acrosome reaction and multivalence GlcNAc activity (sperm of acrosome reaction, %). PAA (GlcNAc; 0.2) when being 100 μ M, GlcNAc concentration induces acrosome reaction significantly (9-17%), and unit price GlcNAc or polymer backbone (pA) are induced the quantity not sufficient mean value (＜5%) of acrosome reaction under the same conditions. The present embodiment explanation, sperm acrosome reaction obviously needs the multivalence of the GlcNAc residue of Gal-reverse transcriptase on the ovum dressing to be combined (from the teeth outwards space clustering), and the synthetic polymer of the derivative that can be connected by the O-that presents a plurality of identical simple GlcNAc brings out.

Fig. 9 a has gathered pAA (GlcNAc; χ) (χ=0.2,0.3,0.6 and 1.0) and pAA (Gal; χ) (χ=0.3,0.6 and 1.0) series induce activity aspect the mouse sperm acrosome reaction (sperm of acrosome reaction, %). Polymer pAA (GlcNAc) shows the active ability weak (4-10%) of regulating when 0.1-100 μ M ([GlcNAc]) concentration. They show increased activity (17-27%) more than 100 μ M. In addition, the result shows that (sperm of acrosome reaction %) increases sharply the activity of pAA when GlcNAc concentration is about 100 μ M (GlcNAc). Fig. 9 a also shows poly-(acrylic acid), the i.e. activity of pAA (Gal) of the galactoside of presenting the O-connection. Low (or without) activity may be since the Gal residue of pAA (Gal) to low (or without) affinity combination of Gal reverse transcriptase.

Fig. 9 b is 100 μ M pAA (GlcNAc; χ) (sperm of acrosome reaction is %) with the drawing of the molar fraction (χ) of the GlcNAc of pAA (GlcNAc) for the activity of (χ=0.2,0.3,0.6 and 1.0). This figure is derived from by Fig. 9 a, and, 14% pAA (NeuAc; 1.0) be estimated value. This has utilized 100 μ M ([GlcNAc]) concentration, because it is to make the active threshold concentration that greatly increases of pAA (GlcNAc). Correlation between activity and the χ is nonlinear; (a) activity increases sharply in the time of between χ=0 and χ=0.2; (b) activity reduces gradually after reaching maximum activity (23%).

Figure 10 has gathered the GlcNAc of multivalence polymerization to the inhibition activity of sperm-ovum combination. Measuring concentration is three kinds of polymer (pAA (GlcNAc of 100 μ M ([GlcNAc]); χ=0.3,0.6 and 1.0)) active to the inhibition of external sperm-ovum combination, three kinds of polymer are reduced to respectively about 10,5 and 5 with the sperm count of each ovum combination from 26 (control group and pA). The result shows that the multivalence GlcNAc of polymerization induces the acrosome reaction of mouse sperm, and suppresses sperm-ovum combination. Polymer backbone, pA are used as control group (concentration 100 μ M, [CONH₂]). Conclusion

(1) pAA (GlcNAc) induces the acrosome exocytosis, and unit price GlcNAc can not; Polyvalency is included in the acrosome process.

(2) nonspecific part is the effectiveness that the pAA (Gal) that passs the Gal reverse transcriptase does not induce the acrosome exocytosis.

It is (3) active that (sperm of acrosome reaction, %) molar fraction (χ) with the pAA (GlcNAc) of some concentration GlcNAc has non-linear relation.

(4) pAA (GlcNAc) suppresses sperm-ovum combination; This mechanism may comprise the sperm acrosome reaction that multivalence GlcNAc induces. Embodiment 5 usefulness semi-solid phase reactions prepare pMVMA (NeuAc) and (see Figure 11 a)

Pass through RNH₂(NeuAc-L ₁NH ₂) prepare methyl vinyl ether maleic acid copolymer (NeuAc-L with methyl vinyl ether maleic anhydride copolymer (or pMVMAn) reaction₁) (or pMVMA (NeuAc-L₁)) copolymer solution, or with the RNH of different molar equivalents (mol equiv)₂Acid anhydrides group (mol equiv={RNH with pMVMAn₂Molal quantity/{ molal quantity of the acid anhydrides group of pMVMAn }) and regulate pH value to 12 with amine aqueous solution. The copolymer pMVMA (NeuAc) of mol equiv＞0 is prepared as follows in little titer plate of 96 circular cone bottom outlets is arranged: (i) with 3mg pMVMAn (M_n=67000,80000,311000,485000 or 1130000gmol^-1) put into the hole; (ii) with the 0.1M RNH of various amounts (10-38mL)₂(NeuAc-L ₁NH ₂) PBS buffer solution (pH12) soak powder in the hole; (iii) seal immediately titer plate, then ultrasonic processing mixture 0.5h. Before the inhibition that carries out the chicken erythrocyte agglutination that influenza virus induces is measured, in the hole that has generated various copolymer solutions (about pH3), add 30mL 1.0M NaOH, solution is transferred to neutral to about pH7, and add to 100 or 200mL (cumulative volume) with PBS (pH7.2). Embodiment 6 usefulness semi-solid phase reactions prepare pMVMA (NeuAc; R) (see Figure 11 b)

Prepare ternary polymer pMVMA (NeuAc-L with similar embodiment 5 used methods₁ R). Ultrasonic processing pMVMAn, NeuAc-L₁NH ₂And R₂NH ₂The mixture of (aliphatic amine, aromatic amine, amino acid, amino sugar or peptide) three kinds of compositions. Mol equiv (mol equiv=(NeuAc-L₁NH ₂And R₂NH ₂The acid anhydrides group of)/pMVMAn)＞0 ternary polymer pMVMA (NeuAc; R₂) in little titer plate of 96 circular cone bottom outlets is arranged, be prepared as follows: (i) with 3mg pMVMAn (M_n=67000,80000,311000,485000 or 1130000gmol^-1) put into the hole; (ii) with 10mL 0.1M NeuAc-L₁NH ₂0.1M RNH with various amounts (2-20mL)₂The PBS buffer solution (pH12) of (for example, naphthyl alanine, phenylalanine, cyclo-hexylamine, phenylethylamine, PABA or mannosamine) soaks powder in the hole; (iii) seal immediately titer plate, then ultrasonic processing mixture 0.5h. Before the inhibition that carries out the chicken erythrocyte agglutination that influenza virus induces is measured, in the hole that has generated various copolymer solutions (about pH3), add 30mL 1.0M NaOH, solution is transferred to neutral to about pH7, and add to 100 or 200mL (cumulative volume) with PBS (pH7.2). Embodiment 7 usefulness semi-solid phase reactions prepare pAA (Gal) (seeing Figure 11 c)

Pass through RNH₂(Gal-β-L ₂NH ₂；Gal-α-L ₃NH ₂) and poly-(acrylic anhydride) (pAAn; M_n＝20700gmol ^-1，M _w＝39500gmol ^-1) reaction preparation pAA (Gal) copolymer solution, with the RNH of different molar equivalents (mol equiv)₂Acid anhydrides group (mol equiv={RNH with pAAn₂Molal quantity/{ molal quantity of the acid anhydrides group of pAAn }) and regulate pH value to 12 with amine aqueous solution. The copolymer pAA (Gal) of mol eqmv＞0 is prepared as follows in little titer plate of 96 circular cone bottom outlets is arranged: (i) 6mg pAAn is put into the hole; (ii) with the 0.1M RNH of various amounts (10-100mL)₂(Gal-β-L ₂NH ₂Or Gal-α-L₃NH ₂) PBS buffer solution (pH12) soak powder in the hole; (iii) seal immediately titer plate, then ultrasonic processing mixture 0.5h. Before the inhibition that carries out the chicken erythrocyte agglutination that influenza virus induces is measured, in the hole that has generated various copolymer solutions (about pH3), add 60mL 1.0M NaOH, solution is transferred to neutral to about pH7, and add to 100 or 200mL (cumulative volume) with PBS (pH7.2). Embodiment 8 usefulness semi-solid phase reactions prepare pBMA (Gal) (seeing Fig. 6 d)

Prepare copolymer butadiene maleic acid copolymer (Gal) (or pBMA (Gal)) with similar embodiment 7 used methods. Pass through RNH₂(Gal-β-L ₂NH ₂；Gal-α-L ₃NH ₂) and butadiene maleic anhydride copolymer (pBMAn; M_w＝10000-15000gmol ^-1) reaction preparation pBMA (Gal) copolymer solution, with the RNH of different molar equivalents (mol equiv)₂Acid anhydrides group (mol equiv={RNH with pBMAn₂Molal quantity/{ molal quantity of the acid anhydrides group of pBMAn }) and regulate pH value to 12 with amine aqueous solution. The copolymer pBMA (Gal) of mol equiv＞0 is prepared as follows in little titer plate of 96 circular cone bottom outlets is arranged: (i) 6mg pBMAn is put into the hole; (ii) with the 0.1M RNH of various amounts (10-100mL)₂(Gal-β-L ₂NH ₂Or Gal-α-L₃NH ₂) PBS buffer solution (pH12) soak powder in the hole; (iii) seal immediately titer plate, then ultrasonic processing mixture 0.5h. Before the inhibition that carries out the chicken erythrocyte agglutination that influenza virus induces is measured, in the hole that has generated various copolymer solutions (about pH3), add 60mL 1.0M NaOH, solution is transferred to neutral to about pH7, and add to 100 or 200mL (cumulative volume) with PBS (pH7.2). Embodiment 9: prepare pAA (SL with semi-solid phase reaction^X _e) (referring to accompanying drawing 12a)

Prepare copolymer pAA (SL with similar embodiment 7 used methods^X _e). With RNH2 (SL^X _e -NH ₂) from the pAAn reaction RNH with different molar equivalents (mol equiv)₂Anhydride group (mol equiv={RNH with pAAn₂Molal quantity/{ the anhydride group molal quantity of pAAn }) and to regulate pH with amine aqueous solution be 12 preparation copolymer pAA (SL^X) solution. Copolymer pAA (the SL of mol equiv＞0^X) in little titer plate of 96 conical bottom outlets, be prepared as follows: (i) with 6mg pAAn (M_n＝20700gmol ^-1，M _w＝39500gmol ^-1) be placed in the hole: (10-100mL) 0.1M RNH that (ii) uses variable₂(SL ^X _eRNH ₂) PBS buffer solution (pH 12) soak powder; (iii) seal immediately plate, then ultrasonic processing mixture is 0.5 hour. Each reactant mixture of the copolymer that will prepare in the hole (pH about 3) is neutralized to pH about 7 and is adjusted to 100 or 200mL (cumulative volume) with PBS (pH 7.2) by adding 60mL 1.0M NaOH, then carries out neutrocyte and endothelial cell adhesion inhibition mensuration. Embodiment 10: prepare pAA (bacitracin with semi-solid phase reaction; R) (referring to accompanying drawing 12b)

Prepare ternary-polymer pAA (bacitracin with similar embodiment 6 used methods; R). To comprise pAAn, bacitracin and R₂NH ₂The ultrasonic processing of ternary mixture of (fatty amine, aromatic amine, amino acids, aminosugars, or peptide class). The ternary of the mol equiv of the acid anhydride base of bacitracin and RNH2 and pAAn＞0-polymer pAA (bacitracin; R) in little titer plate of 96 conical bottom outlets, be prepared as follows: (i) with 3mg pAAn (M_n＝20700gmol ^-1，M _w＝39500gmol ^-1) be placed in the hole: (2-20mL) 0.1M RNH that (ii) uses 10mL0.1M bacitracin and variable₂The PBS buffer solution (pH 12) of (example: naphthyl alanine, phenylalanine, cyclohexylamine, phenyl ethylamine, PABA, or mannosamine) soaks powder; (iii) seal immediately plate, then ultrasonic processing mixture is 0.5 hour. Each solution of the ternary-polymer that will prepare in the hole (pH about 3) is neutralized to pH about 7 and is adjusted to 100 or 200mL (cumulative volume) with PBS (pH 7.2) by adding 30mL 1.0M NaOH, then carries out bacterium growth inhibition mensuration. Equivalent

Those skilled in the art will be familiar with, or can only determine many equivalents of instantiation described here and method with normal experiment. These equivalents will be included in the following claim scope.

Claims

1. preparation method that the multivalence that is used for patient treatment disease or indication is presented thing comprises:

(a) making up and arrange a plurality of group A forms on framework in order to treat disease and suitable

Answer the multivalence of disease to present thing, described structure and arrangement are to be based upon because described base

The multivalence of the A of group and target binding site B set interacts and has covered in the patient

Target binding site B gathers and produces on the basis of therapeutic action, wherein said covering

Lid is presented by a plurality of multivalence optionally that thing and target binding site B gather

Conformal interfacial interaction causes;

(b) a plurality of group A make up and arrange at framework, and described method optionally

Also comprise one or more following steps: this structure and arrangement are based upon multivalence and are

Pass thing and form the ability of gel sample barrier around the target binding site B array

On, wherein barrier is to present being total to of thing and target binding site B set by multivalence

The shape interfacial interaction forms;

(c) make up and arrange a plurality of group A at framework, this structure and arrangement are based upon many

Valency is presented steric target known or expection on thing and the patient's inner boundary

On the interactional ability of binding site B set;

(d) based on to the interaction between separate base A and a plurality of target binding site B

Analysis select the weak group A of a kind of binding ability, condition is a plurality of group A

Arrive set in conjunction with threshold, wherein in conjunction with being to present thing by multivalence to be combined the position with target

The conformal interfacial interaction of some B set forms;

(e) make up and arrange a plurality of group A at framework, this structure and arrangement be based upon to

Multivalence is presented thing and is given on the flexible basis, and itself and target binding site B are gathered

Consistent;

(f) select a kind of framework, can give multivalence with required characteristic and present thing,

Wherein said required characteristic is selected from arbitrarily multivalence and presents the flexibility of thing and solvable

The raising of property.

2. the process of claim 1 wherein that a plurality of group A stick on the framework by attachment, described method is optional also to comprise one or more following steps: (a) give the type that ability that multivalence presents thing is selected attachment according to attachment with flexibility; (b) according to attachment group A is offered the hit ability of binding site B set of patient and select the type of attachment; (c) group A is offered the hit factor of ability of binding site B set of patient select length and the type of attachment according to affecting attachment, described factor is selected from arbitrarily hydrophobicity, hydrophily, the diameter of attachment, the hydrophobicity of wherein said attachment or hydrophilic selection are hydrophobicity or the hydrophilies according to patient's vivo environment, condition is can be near target binding site B, and the selection of described attachment diameter is according to the known of patient body internal channel or expection diameter, and condition is can be near target binding site B.

3. method for the treatment of disease or indication, comprise: use a plurality of group A to the patient, choose wantonly and present the thing form with multivalence, present the conformal interfacial interaction realization of binding site B set in thing and the patient body to the treatment of disease or indication by multivalence, conformal interfacial interaction is covered set or the array of the target binding site B in the patient body.

4. many group A, the optional multivalence that adopts is presented the thing form, for the preparation of the purposes in the medicine for the treatment of disease or indication in individuality, wherein be to present the conformal interfacial interaction realization disease of the target binding site B set in thing and the patient body or the treatment of indication by multivalence, conformal interfacial interaction is covered set or the array of the target binding site B in the patient body.

5. many group A, wherein the optional multivalence that adopts is presented the thing form, be used for the treatment of disease or indication, be used for patient's administration, present the conformal interfacial interaction realization disease of the target binding site B set in thing and the patient body or the treatment of indication by multivalence, conformal interfacial interaction is covered set or the array of the target binding site B in the patient body.

6. method for the treatment of disease or indication, comprise: use a plurality of group A to the patient, interact and realize treatment to disease or indication so that present a plurality of binding site B in thing and the patient body by multivalence, described multivalence is presented thing and is contained a plurality of group A that adhere on the framework, and described multivalence is presented thing and met the following requirements: i) group A is functional group and plays drug effect, can unite alone or with framework

Use; Ii) the group A that adheres on the framework will be presented one, preferred 2 or 3 at least

Individual, attendant advantages is for interaction, with respect to separate base A to some

Target binding site B presents; Iii) this attendant advantages is collaborative advantage, and this advantage is than the phase that is dispersed in the homogeneous solution

Want large with the attendant advantages that the set of group A monomer may provide; And wherein said attendant advantages is selected from arbitrarily by low concentration group A sufficient biological effect is provided, and improves the selectivity for target and non-target site, in conjunction with aspect positive cooperativity, in conjunction with the entropy rising and have the preparation of presenting thing of low dissociation yield.

7. many group A are for the preparation of the purposes in the medicine for the treatment of disease or indication, interact and realize treatment to disease or indication so that present a plurality of binding site B in thing and the patient body by multivalence, described multivalence is presented thing and is contained a plurality of group A that adhere on the framework, and described multivalence is presented thing and met the following requirements: i) group A is functional group and plays drug effect, can unite alone or with framework

Individual, attendant advantages is for interaction, with respect to separate base A to a plurality of

Want large with the attendant advantages that the set of group A monomer may provide; And

Wherein said attendant advantages is selected from arbitrarily by low concentration group A sufficient life is provided

The thing effect improves the selectivity for target and non-target site, in conjunction with aspect just association

The same sex, in conjunction with entropy raise and have the preparation of presenting thing of low dissociation yield.

8. a plurality of group A that are used for the treatment of the interior disease of patient body or indication, give by this way patient's administration so that present in thing and the patient body some the binding site B treatment of realizations to disease or indication that interact by multivalence, described multivalence is presented thing and is contained a plurality of group A that adhere on the framework, and described multivalence is presented thing and met the following requirements: i) group A is functional group and plays drug effect, can unite alone or with framework

9. a method for the treatment of disease or indication comprises: use two kinds of dissimilar group (A to the patient at least₁And A₂), present the interaction realization of binding site B set in thing and the patient body to the treatment of disease or indication by multivalence, described multivalence is presented thing and is contained group (A at framework₁And A₂), wherein, group A₁And A₂One of can not be carbohydrate.

10. at least two kinds of dissimilar group (A₁And A₂) for the preparation of the purposes in the medicine for the treatment of disease or indication, wherein said group (A₁And A₂) be used to the patient and then present the interaction realization of binding site B set in thing and the patient body to the treatment of disease or indication by multivalence, described multivalence is presented thing and is contained group (A at framework₁And A₂), wherein, group A₁And A₂One of can not be carbohydrate.

11. at least two kinds of two dissimilar group (A₁And A₂), be used for the treatment of disease or indication, wherein said group is used to the patient and then presents the interaction realization of binding site B set in thing and the patient body to the treatment of disease or indication by multivalence, and described multivalence is presented thing and contained group (A at framework₁And A₂), wherein, group A₁And A₂One of can not be carbohydrate.

12. a method for the treatment of disease or indication comprises: use at least three kinds of dissimilar group (A to the patient₁，A ₂And A₃), present the interaction realization of binding site B set in thing and the patient body to the treatment of disease or indication by multivalence, described multivalence is presented thing and is contained group (A at framework₁，A ₂And A₃)。

13. at least three kinds of dissimilar group (A₁，A ₂And A₃), for the preparation of the purposes in the medicine for the treatment of disease or indication, and then present the interaction realization of binding site B set in thing and the patient body to the treatment of disease or indication by multivalence, described multivalence is presented thing and is contained group (A at framework₁，A ₂And A₃)。

14. at least three kinds of dissimilar group (A₁，A ₂And A₃), being used for the treatment of disease or indication, described group is used to the patient and then presents the interaction realization of binding site B set in thing and the patient body to the treatment of disease or indication by multivalence, and described multivalence is presented thing and is contained group (A at framework₁，A ₂And A₃)。

15. method that promotes to treat disease or indication, comprise: use some group A to the patient, so that present the interaction promotion of binding site B set in thing and the patient body to the treatment of disease or indication by multivalence, described multivalence is presented thing and is contained group A at framework, and wherein, group A is not carbohydrate, the carbohydrate derivative, antibody or its segment, Y is not glucan, and multivalence to present thing be not vaccine; Or group A is not carbohydrate or PB.

16. some group A are for the preparation of the purposes in the medicine for the treatment of disease or indication, so that present the interaction promotion of binding site B set in thing and the patient body to the treatment of disease or indication by multivalence, described multivalence is presented thing and is contained group A at framework, wherein, group A is not carbohydrate, carbohydrate derivative, antibody or its segment, Y is not glucan, and multivalence to present thing be not vaccine; Or group A is not carbohydrate or PB.

17. some group A, be used for the treatment of disease or indication, give treatment patient's administration so that present the interaction promotion of binding site B set in thing and the patient body to the treatment of disease or indication by multivalence, described multivalence is presented thing and is contained group A at framework, and wherein, group A is not carbohydrate, the carbohydrate derivative, antibody or its segment, Y is not glucan, and multivalence to present thing be not vaccine; Or group A is not carbohydrate or PB.

18. according to claim 3, one of 6,9,12 and 15 method, described method comprises following one or more optional variation: (a) use to adhere to as multivalence and present group A on the framework of a part of thing; (b) multivalence is presented thing and is assembled in vivo; (c) a plurality of target binding site B appear on the same interface; (d) a plurality of target binding site B appear on the different interfaces; (e) target binding site B comprises more than one binding sites; (f) present the non-single-minded combination that weakens that thing shows group A and non-B binding site with respect to multivalence for the single-minded combination of binding site B; (g) multivalence of the group A of low concentration is presented thing to be than group A that the unit price of identical molar concentration is presented group A more effective; (h) space suppresses, and optional noncompetitive is used for improving result for the treatment of; (i) the group A that presents of multivalence has the higher affinity of group A of presenting than unit price for target binding site B; (j) multivalence is presented the body gathering that thing mediation target binding site B exists; (k) the multivalence framework of presenting thing contains non-covalent bond, the optional component that is selected from following material that comprises of described framework: liposome, liposome derivative, micelle, colloid, protein aggregate and cell; (l) the multivalence framework of presenting thing contains covalent bond, the optional biological particle that is selected from following material that comprises of described framework: sugar, protein, lipid and little molecular biosciences particle; (m) the multivalence framework of presenting thing contains attachment, and optional acid amides connects thing; (n) framework comprises the chemical part that is selected from following group: hydrolyzable attachment, dendrimer, polymer, choose wantonly and formed by the following member: poly-(ester), poly-(acid anhydrides), carbohydrate, how pure, poly-(acrylate), poly-(methacrylate), poly-(ether) and poly-(amino acid), poly-(glutamic acid), poly-(aspartic acid), glucan, dextran sulfate, poly-(maleic anhydride-altogether-vinyl ethers), poly-(succinimide), poly-(acrylic anhydride), PEG, poly-(lactic acid), poly-(glycolic acid), PVP, poly-(phenylethylene-maleic anhydride), poly-(alpha-olefin-maleic acid), hyaluronic acid, sodium carboxymethylcellulose, chondroitin sulfate, poly-(acrylamide), poly-(glycerine), starch and their derivatives, copolymer and homopolymers, described polymer is optional to link to each other with group A by covalent bond; (o) to present thing be water miscible to described multivalence, chooses wantonly in mg/ml scope or grams per milliliter scope; (p) to present the volume that thing distributes in patient body be predictable to multivalence; (q) multivalence is presented the molecular weight of thing greater than 60kD; (r) the multivalence character of presenting thing is better than the group A that unit price is presented, and described character refers to action time or therapeutic index; (s) multivalence is presented thing has the longer half-life at some special parts than group A that unit price is presented in patient body, as at blood, and central nervous system, lung is in GI road and the kidney; And (t) multivalence present the average hydrodynamics diameter of thing greater than 50 dusts.

19. claim 4, the purposes of one of 7,10,13 and 16 group A, described purposes comprises following one or more optional variation: (a) use to adhere to as multivalence and present group A on the framework of a part of thing; (b) multivalence is presented thing and is assembled in vivo; (c) a plurality of target binding site B appear on the same interface; (d) a plurality of target binding site B appear on the different interfaces; (e) target binding site B comprises more than one binding sites; (f) with respect to for the single-minded combination of binding site B, multivalence is presented the non-single-minded combination that weakens that thing shows group A and non-B binding site; (g) multivalence of the group A of low concentration is presented thing to be than group A that the unit price of identical molar concentration is presented group A more effective; (h) space suppresses, and optional noncompetitive is used for improving result for the treatment of; (i) the group A that presents of multivalence has the higher affinity of group A of presenting than unit price for target binding site B; (j) multivalence is presented the body gathering that thing mediation target binding site B exists; (k) the multivalence framework of presenting thing contains non-covalent bond, the optional component that is selected from following material that comprises of described framework: liposome, liposome derivative, micelle, colloid, protein aggregate and cell; (l) the multivalence framework of presenting thing contains covalent bond, the optional biological particle that is selected from following material that comprises of described framework: sugar, protein, lipid and little molecular biosciences particle; (m) the multivalence framework of presenting thing contains attachment, and optional acid amides connects thing; (n) framework comprises the chemical part that is selected from following group: hydrolyzable attachment, dendrimer, polymer, choose wantonly and formed by the following member: poly-(ester), poly-(acid anhydrides), carbohydrate, how pure, poly-(acrylate), poly-(methacrylate), poly-(ether) and poly-(amino acid), poly-(glutamic acid), poly-(aspartic acid), glucan, dextran sulfate, poly-(maleic anhydride-altogether-vinyl ethers), poly-(succinimide), poly-(acrylic anhydride), PEG, poly-(lactic acid), poly-(glycolic acid), PVP, poly-(phenylethylene-maleic anhydride), poly-(alpha-olefin-maleic acid), hyaluronic acid, sodium carboxymethylcellulose, chondroitin sulfate, poly-(acrylamide), poly-(glycerine), starch and their derivatives, copolymer and homopolymers, described polymer is optional to link to each other with group A by covalent bond; (o) to present thing be water miscible to described multivalence, chooses wantonly in mg/ml scope or grams per milliliter scope; (p) to present the volume that thing distributes in patient body be predictable to multivalence; (q) multivalence is presented the molecular weight of thing greater than 60kD; (r) the multivalence character of presenting thing is better than the group A that unit price is presented, and described character refers to action time or therapeutic index; (s) multivalence is presented thing has the longer half-life at some special parts than group A that unit price is presented in patient body, as at blood, and central nervous system, lung is in GI road and the kidney; And (t) multivalence present the average hydrodynamics diameter of thing greater than 50 dusts.

20. according to claim 5, one of 8,11,14 and 17 group A, described treatment comprises following one or more optional variation: (a) use to adhere to as multivalence and present group A on the framework of a part of thing; (b) multivalence is presented thing and is assembled in vivo; (c) a plurality of target binding site B appear on the same interface; (d) a plurality of target binding site B appear on the different interfaces; (e) target binding site B comprises more than one binding sites; (f) with respect to for the single-minded combination of binding site B, multivalence is presented the non-single-minded combination that weakens that thing shows group A and non-B binding site; (g) multivalence of the group A of low concentration is presented thing to be than group A that the unit price of identical molar concentration is presented group A more effective; (h) space suppresses, and optional noncompetitive is used for improving result for the treatment of; (i) the group A that presents of multivalence has the higher affinity of group A of presenting than unit price for target binding site B; (j) multivalence is presented the body gathering that thing mediation target binding site B exists; (k) the multivalence framework of presenting thing contains non-covalent bond, the optional component that is selected from following material that comprises of described framework: liposome, liposome derivative, micelle, colloid, protein aggregate and cell; (l) the multivalence framework of presenting thing contains covalent bond, the optional biological particle that is selected from following material that comprises of described framework: sugar, protein, lipid and little molecular biosciences particle; (m) the multivalence framework of presenting thing contains attachment, and optional acid amides connects thing; (n) framework comprises the chemical part that is selected from following group: hydrolyzable attachment, dendrimer, polymer, choose wantonly and formed by the following member: poly-(ester), poly-(acid anhydrides), carbohydrate, how pure, poly-(acrylate), poly-(methacrylate), poly-(ether) and poly-(amino acid), poly-(glutamic acid), poly-(aspartic acid), glucan, dextran sulfate, poly-(maleic anhydride-altogether-vinyl ethers), poly-(succinimide), poly-(acrylic anhydride), PEG, poly-(lactic acid), poly-(glycolic acid), PVP, poly-(phenylethylene-maleic anhydride), poly-(alpha-olefin-maleic acid), hyaluronic acid, sodium carboxymethylcellulose, chondroitin sulfate, poly-(acrylamide), poly-(glycerine), starch and their derivatives, copolymer and homopolymers, described polymer is optional to link to each other with group A by covalent bond; (o) to present thing be water miscible to described multivalence, chooses wantonly in mg/ml scope or grams per milliliter scope; (p) to present the volume that thing distributes in patient body be predictable to multivalence; (q) multivalence is presented the molecular weight of thing greater than 60kD; (r) the multivalence character of presenting thing is better than the group A that unit price is presented, and described character refers to action time or therapeutic index; (s) multivalence is presented thing has the longer half-life at some special parts than group A that unit price is presented in patient body, as at blood, and central nervous system, lung is in GI road and the kidney; And (t) multivalence present the average hydrodynamics diameter of thing greater than 50 dusts.

21. according to claim 3,6,9, one of 12 and 15 method, wherein, multivalence is presented thing and is prevented or suppress following interaction: cell-cell interaction, and amphigamy, cell-pathogen interacts, pathogen-extracellular matrix interacts, pathogen-extracellular matrix interacts, and neutrophil leucocyte-endothelial cell interacts, inflammation, metastasis of cancer and blood platelet-blood platelet interacts, wherein said multivalence presents that thing is optional adjusts the biological part that cell migration or blocking-up are selected from integrin and platelet receptor, and described method optional being used for the treatment of is selected from Acute thrombosis, hypocoagulability and hypercoagulable indication.

22. according to claim 4,7,10, the purposes of one of 13 and 16 group A, wherein, multivalence is presented thing and is prevented or suppress following interaction: cell-cell interaction, amphigamy, cell-pathogen interacts, pathogen-extracellular matrix interacts, pathogen-extracellular matrix interacts, and neutrophil leucocyte-endothelial cell interacts, inflammation, metastasis of cancer and blood platelet-blood platelet interacts, wherein said multivalence presents that thing is optional adjusts the biological part that cell migration or blocking-up are selected from integrin and platelet receptor, and described method optional being used for the treatment of is selected from Acute thrombosis, hypocoagulability and hypercoagulable indication.

23. according to claim 5,8,11, one of 14 and 17 group A, wherein, multivalence is presented thing and is prevented or suppress following interaction: cell-cell interaction, and amphigamy, cell-pathogen interacts, pathogen-extracellular matrix interacts, pathogen-extracellular matrix interacts, and neutrophil leucocyte-endothelial cell interacts, inflammation, metastasis of cancer and blood platelet-blood platelet interacts, wherein said multivalence presents that thing is optional adjusts the biological part that cell migration or blocking-up are selected from integrin and platelet receptor, and described method optional being used for the treatment of is selected from Acute thrombosis, hypocoagulability and hypercoagulable indication.

24. method that suppresses gestation, comprise: use a plurality of group A to the patient, the optional GlcNAc that contains of group A wherein, and then present thing and a plurality of target binding site B by multivalence and interact realization to the inhibition of gestation, described multivalence is presented thing and is contained group A at framework.

25. many group A, purposes in the medicine pregnant for the preparation of inhibition, comprise: use a plurality of group A to the patient, wherein group A chooses wantonly and contains GlcNAc, and then present thing and a plurality of target binding site B by multivalence and interact and realize inhibition to gestation, described multivalence is presented thing and is contained group A at framework.

26. be used for to suppress a plurality of group A of gestation, wherein group A is optional contains GlcNAc, after using to the patient, presents thing and a plurality of target binding site B by multivalence and interacts realization to the inhibition of gestation, described multivalence is presented thing and is contained group A at framework.

27. claim 3, one of 6,9,12 and 15 method, wherein, multivalence is presented thing can prevent following biological interaction: cell-pathogen adhesion and pathogen-extracellular matrix are adhered, and wherein pathogen is selected from arbitrarily bacterium, fungi, virus and parasite.

28. claim 4,7,10, the purposes of one of 13 and 16 group A, wherein, multivalence is presented thing can prevent following biological interaction: cell-pathogen adhesion and pathogen-extracellular matrix are adhered, wherein pathogen is selected from arbitrarily bacterium, fungi, virus and parasite.

29. claim 5, one of 8,11,14 and 17 group A, wherein, multivalence is presented thing can prevent following biological interaction: cell-pathogen adhesion and pathogen-extracellular matrix are adhered, and wherein pathogen is selected from arbitrarily bacterium, fungi, virus and parasite.

30. a multivalence is the pharmaceutical composition of delivery of therapeutic agents, and contain the multivalence that can be represented by the formula and present thing,

(Y)-(X-A) _n

Wherein, Y is framework, optionally is aggregation framework, and X is straight key or linking group,

Described linking group is optional to be the part of independent sector rather than Y or A, the functional group that A is presented, and n is the integer greater than 10, and optional also comprise one or more following variations: (a) described integer is selection like this, and (X-A) part adhere on the Y, be consistent with target binding site B set when being used for the treatment of the patient so that multivalence is presented thing, and when being used for the patient, choose wantonly and cover target binding site B array; (b) described integer is selection like this, is consistent with target binding site B set when being used for the treatment of the patient so that multivalence is presented thing; Reaching (c), described composition also contains arbitrarily pharmaceutically suitable carrier.

31. consistent multivalence is presented thing, contains to form multivalence and present a plurality of group A on the framework of thing, it is so consistent that described multivalence is presented thing, so that it has covered the target binding site set in the patient body.

32. a pharmaceutical composition contains at least two kinds of different multivalence and presents thing and pharmaceutically suitable carrier.

33. the method for adhesion effect between the biomolecule of a plurality of bonding surfaces of adjustment and a plurality of the second molecule, comprise: the combining form of presenting thing with the biomolecule of a plurality of bonding surfaces and multivalence provides, described multivalence presents that thing is optional to be non-carbohydrate and raising factor-beta greater than about 100 is arranged, contain a framework, a plurality of the third molecules in adhesion on the framework, wherein said the third molecule is by on the biomolecule that simultaneously described the third molecule is bonded to a part of described bonding surface and carry out the space by the described framework of biomolecule with the described bonding surface of a part of the third molecule that do not bond and block to adjust the biomolecule of described bonding surface and the adhesion effect between described the second molecule, wherein said adhesion effect is optional to be occurred in cell and is selected from bacterium, virus, influenza virus is between the entity of the second cell and multivalent molecule.

34. multivalence is presented thing, wherein is selected from: pAA (Gal-β), pAA (Gal-α), pBMA (Gal-β), pBMA (Gal-α) and pAA (GlcNAc-β).

35. one kind prevents the ricin erythrocytic method of adhering, comprise described ricin and effective dose be selected from pAA (Gal-β), pAA (Gal-α), pBMA (Gal-β) presents thing with the multivalence of pBMA (Gal-α) and contacts.

36. one kind prepares pAA (Gal-β) or pBMA (Gal-β) multivalence is presented the method for thing, comprising: (a) with Gal-β O-L₁NH ₂With poly-(N-acryloyl group succinimide) or poly-(fourth

Diene-altogether-maleic anhydride) reaction, and (b) the described reaction of cancellation.

37. one kind prepares pAA (Gal-α) or pBMA (Gal-α) multivalence is presented the method for thing, comprising: (a) with Gal-α C-L₂NH ₂With poly-(N-acryloyl group succinimide) or poly-

(butadiene-altogether-maleic anhydride) reaction, and (b) the described reaction of cancellation.

38. a method that suppresses individual fertilization comprises that pAA (GlcNAc-β) multivalence of using effective dose to described individuality presents thing.

39.pAA (GlcNAc-β) multivalence is presented thing, the purposes in the composition of being fertilized for the preparation of inhibition.

Present thing 40. be used for to suppress the multivalence of fertilization, described multivalence is presented thing to be pAA (GlcNAc-β) and to use to individuality by suppressing the fertilization effective dose.

41. one kind prepares pAA (GlcNAc-β) multivalence and presents the method for thing, comprising: (a) with GlcNAc-β-L₁NH ₂Anti-with poly-(N-acryloyl group succinimide)

Should, and (b) the described reaction of cancellation.

42. make the method that multivalence is presented the thing array, comprising for one kind: (a) the first multivalence is presented the frame part of first activation of thing and the frame part that the second multivalence is presented first activation of thing and be added in the first and second reaction vessels; (b) the first multivalence being presented first functional group part of thing and first functional group that the second multivalence is presented thing partly is added in described the first and second reaction vessels; (c) allow the composition of activation to react so form the array that at least two kinds of different multivalence are presented thing; The method is chosen wantonly and also comprised one or more the following steps: first interval group of (d) described the first multivalence being presented thing is added in described the first reaction vessel; (e) first interval group of described the second multivalence being presented thing is added in described the second reaction vessel; (f) auxiliary group described the first multivalence being presented thing is added in described the first reaction vessel; (g) auxiliary group described the second multivalence being presented thing is added in described the second reaction vessel; The method is optional also to comprise following one or more variation: (l) repeat above-mentioned step, wherein " n " individual described the first and the second multivalence functional group of presenting thing is added to respectively in described the first and second reaction vessels; (2) repeat above-mentioned step, wherein " n " individual described the first and the second multivalence interval group of presenting thing is added to respectively in described the first and second reaction vessels; (3) repeat above-mentioned step, wherein " n " individual described the first and the second multivalence auxiliary group of presenting thing is added to respectively in described the first and second reaction vessels.

43. useful claim 1, the multivalence of one of 2,36,41 or 42 method preparation presents thing or multivalence is presented the thing array.