SE9400450A0

SE9400450A0 - Preparation and application of artificial anti-idiotypic antibodies obtained by molecular imprinting

Info

Publication number: SE9400450A0
Application number: SE9400450A
Authority: SE
Inventors: Klaus Mosbach
Original assignee: Klaus Mosbach
Priority date: 1994-02-10
Filing date: 1994-02-10
Publication date: 1995-08-11
Also published as: DE69514785T2; ATE189130T1; SE9400450D0; DK0743870T3; EP0982591A3; EP0743870B1; EP0982591A2; AU1828095A; JPH09510699A; US6489418B1; DE69514785D1; WO1995021673A1; EP0743870A1

Description

to FEB. *'94 1s=52 mm- Bloc tunn +4s-4ß~1n ïffflft *ïil-.fïšß- in obtnittetl tnolecnle, R' which could be called "ﬁllt;r", shnuld resemble or be complemcntary to the original itnprint :nolecnle in shape and functionality. but is. dependßnt on the 'ﬂlling ntaterinl' applied. dlffcmnt in contpøsitioxt titan R.

As an extension of the technique one can envlsage. what could lx called surface-imprintittg, the following: (figure 3). A molecular intprint is made against a cotnponnd. ag. compountl X cartwing positive chnrgea. A fnnctionalizetl mntxix (M) caxrying poiymerinaltle groups together with complementary monomers. ng. negatively nhnfged monument. are utllized in the polymetizntion step A. Extraction of the print ninlecnle (fi) leaves the polymer with recognition sites onmplernentary in shape and functionality to componnrl X. Using these sites as ntoltls ut cnsts in a kind nf anti-idiotype polymerization utllizliig complementary monomers, e.g. positively charged monorners, (C) and subsoquettt æmoval of the first polymer (D) mndcrs nn attti-idiotype imprhit polymet' (Y) mirnickittg the original cotnpound X. Thns, the surface-itnprint of the original molecnle (X), is preparat! which could be employetl to create the sttnctnrally related molecule (Y). 'fhls technique should be especially useful for larger molecules.

One xeqnirement for the envisaged ímprint is that both the oxiginnl mattix as well as the subsequetntly obtained matrix are capable of allowing imprittts to bn malin. Another mqnircment is that the original, Le.. the. first itnprixtt ntaterial. can be :anime-A without intcrfezizig with the memorlzirtg cnpability of tha second itnptirit. A number of possible materials and approaches am given in the examples. 'finess include the use of :evefsibly crosslinkirlg monomers such as the inse of ctossIinknrs containing Schlffs base-linkages [6].

The letter polymers m: easily dlssolvetl.

(Ifossllnkexs containing šhiffs bawlinkages IE] FEB. '94 15:53 HPPL BIÜC LUND +46~46°-1B4611 PÛÜE ...an LW .IF l .- 3 Q Another possibility is the use of disulﬁde containing analoga of bls-eorylamide. eg. bis- acrylylcystamine, which can be dissolved with Z-mercaptoetltttnol (71.

H N \,,/\ O QïlšA/KS/w* Bis-acqflylcystarnlno Other crosslinkers that can be cleaved are NN-diallyltartardiantide [8] or N,N'-(l.2- dihydroxyethyleneﬂalsacrylamide [9].

H \/\,,/W\Ä\(§\/\ H OH Û NN-díallyltartardlaxnide N.N'-(1,2-Dihyd:oxy)-bisacrylanxide Another possible way is to use completely different matrices as furst or second imprints such as agaxose or silica, the latver preparcd by polymexization of sllanes. As an alternative co the removal of the first imprint by subsequent dissolution of the matrix. the second imprint may be loosened up using such reversibly dissolving monomers whereby care is to be taken to protect the recognizing properties of the second imprint matrix. Other approaches facllitating such dissociation include the use of magnetic beads canyíng the lmprint molecules. Such matzices can. posterior to polymerization, be scparatcd by application of a magnetic ﬁeld forcing the matrices apart. Another altemative approach lnvolves the polymerlzation at intorfaces. to ren. '94 15:53 nPPL aioc LUND Ms-as-itmett .i PfifiE i '_ '“ iíïåfï: IÜ An additional altemative way to obtain lmprints would be to directly make itnptints of biomolecules by ﬁlling the active or binding sites of the latter with monomets and subsequently using the fornied matrix as a mold or east in a second imprinting step. This would lead to the formation of anti-idiotype impnnts of the first preformed itnprints as depicted in figure 4. The particular binding species is itnmobllined on a degadable matrix (1). eg. agarose beads, and a polytnerization mixture containing functicnally complementary monomers is added in step A. Following polymetization the original matrix. is dissolved in step B making the newly formed polymerlc anti-idiotypic binding sites (3) accessible. Such preparations could then be used e. g. as artiñcial inhibitors or receptors either while arranged as a film or thin membrane or in smaller units. Further. as indicated in figure 4, one can go from 3 over step C employing the imprint of 3 to obtain via step D a plastie imprint 4 similar to 1. Further, as an altemative to the above direct imprlnting, one can envisage the lining up of monomers or other molecules along a surface or active site of egblomolecules as outlined in figure 5. The former are allowed to intet-act with functional groups of 'the rnoleeule, l, in step A followed by their condensation (B). Removal of 1 in step C leads to the formation of a thin-layer imprint of l. Alternatively, the functional groups of the biomolecule are first detivatized followed their conclensatlon.

Potential use of such lmprints (e. filler molecule) (l) They could lead to :new enzyme inhibitors, new drugs. (2) 'lltcy could replace pcptides. nucleoticles, carbohydrates or other biological material with other material such as organic polymets. (3) They could lead to new compounds with identical or similar function as the imprint species but being more stable and cheaper to prepare. (4) In an extension: in cavities or on surfaces obtained by imprinting new enzyme- like catalysts may be obtained. (5) Such cavities if they ate chiral can be used ss scaffold to create new chiral molecules. u: Fas. '94 15: 54 QPPL Bloc LUND +4e-4s~w4s11 A hm X ä 12:* ”u” (6) The cavities obtained from the first imprim can be used m man new molecules uülizing cømbínatoﬁal libraries of vañous organic or inorganic mølccules. (7) Analogous to anxi-idiotypc amibodics.

References 1. N. K. Jema. Ann. Immunoh 125, 373-378 (1974). 2. L. Izadyar, A. Friboulet. M. H. Remy, A. Roseto, D. Thomas, Prac. Narl. Acad.

Scí. USA 90, 8876-8880 (1993). 3. K. Mosbach, U.S. Patent 5.110.833 . (1992). 4. B. Ekberg, K. Mosbach, Trends in Biotechnology 7, 92-96 (1989). 5. G. Vlatakis, L. I. Andersson, R. Müller, K. Mosbach, in Nature. 1993), 645- 647. 6. H. Ringsdorf, G. Greber, Makramol. Chem. 2-5, 237-239 (1958). 7. J. N. Hansen, Anal. Btochem. 76, 37-44 (1976). 8. P. I. Spacth, H. Koblet, Anal. Bíochem. 93, 275-285 (1979). 9.

\'*^ P. B. H. O'Conne1l. C. J. Brady. Anal. Blochem. . 63-73 (1976).

PRGE

Claims

n cøno lß FEB '94 15:31 10 15 20 25 30 35 QNRPHTENT 5.2 CLÅIMS

1. A method ot producing a mimio of en artišioiol or nnuvn ann-uy, o n n r n o 1: e r i n o d in nolymeriinn in a first step mononsra and orosslinkars in the pznsnnoø of the ontity, tharaby fozming a matrix with an imprint of the antity, aøparntinç une untity :rom cha matrix, poly- marining in n aooond stop monomars and croillinknrß. whidh could be tha same nu or dišfnrant from the monomors and oroeniinnern in the first step, in the inprint in the matrix, theraby creating a mimiu of the antity having thi name ﬂunotionnlity and/or nhnpo nn the antity. And thlﬂ aaparating the mimio from the imprint in the matrix.

2. A method of producing a mimio of surface bound molooulas, o h a r a o t e r i s o 4 in poiymeriøing in a titan atep monomazø and oronslinkazø undar the formation of e film having inprinta ot the surface bound moloouløø, soyarating the imprintod film, polymnriling in a second stop monomnrø and orosnlinkazs, which could bn tha namn an oz different írom tha monomars and øroøølinkaro in the first step, on the imprintnd film, tharoby oronting a mimio of tha surface bound molooulon with tha lama tuna- eionality and/or shape as the surface bound molaoulec, and than removing the imprinted film ﬁzom tha mimio.

3. A method of producing a mimio of surfnon bound molnouloa, o h a r a o t 0 r i 0 n d in polymnrisinq monomarn and oronelinkora unna: the formation ot n tilm having imprintn of the surznon bound moloouioo, and than senare-sing :nu inprincad film from the nurﬁnca bound moiooulnn.

4. , 4. Unn of the technique of molooulnr imprinting for tha pzaparßtiün of &nt1~idiotypu lika imprints of mcities oomprinina nztitioinl or natural bindina sites. such nu moleoulnr imprinta, raoeptora, antibodiaa, ennymoo and othara, which anti-idiotypo lika imprinta having the same funccionaiity and/or snaps en tha moitioa. a IDO v v u O O O OO! o 0000 o 15 FEB '94 3! 10 15 20 25 c h a r a c t e r 1 s a d 15131 QNHPQTENT Ü* 7 P'“^“'”;É 5.3

5. Use of molooulor impranting for tha prepßrotion of ant;-idiotypo like molooulon by the polymorioatton Of added monomore and orosnlinkazs within oavitxoa mode by moleoulor tmprincing of moàtåon oomprzøing artitioial or natural binding sites, wheroby the added polymeriaoblo monomora and orosslinkoza orient thomlolvoo within the sites prior to polymorisation.

6. Usa according to oloim B, whoroin tha polymarin- able monomarø era aorylio monomora, nilanas, or other intazoonneating molooules, nuoh aa azidos.

7. Uno of moloculor imprinting for tha proparation of ant1~åd10typø lika impzintß for proparing now moleouløﬂ by their direct formation in or around biomolooulao or other moloculeo utilining preforantiolly polymeriaoblo monomoxs and oronslinkars.

8. Uno of such anti-idzotypa lika imprints oo pzopor- od according to any one of clalml 4-7 96 nßw Lnhibitorl. now drugs. new affinxty ligandn, new anti-atfinity muta- rial including call-atfinity material, anti-antibodioø and now cotolynto.

9. Uno of molooula: impránting for tha praparation of anti-idiotypo imprintm to oraota ohzrality within ohiral oavitieo from non-ohiral baoia alomanto or using ohirol monomera.

10. Uno of a mimio produced oooord¿ng olatm 3 ao o new inhibitor, now drug, now añfinity ligand, new anti- -offinity material including call-otfinity material, anti- -antiboaiaa and now ootalvots.

11. Method of producing an entity compïementary to ß m018CU1@» in adding to the molecuïe. C0mPY1Sfﬂ9 active of binding sites. sma11 cross11nkab1e moities hav1n9 l°t1Ve groups compïementary to the sites on the moïecule. whereby the Smﬂïï moqtqes a$;°c1ate with the above sites, thereafter cross11nk1ng the sma11 moities associated with these sites under the format1on of än entﬂty hqv1nq act1ve groups compïamentary to the moïecuïe. and t en separating the antity from the mo1ecu1e. 10 FEB.'94 15:54 QPPL BIÜC LUND +46-46-104611 PQÛE Summary This patent application describes the use of molecular imptinting as a means for preparation of anti-idiotypic imprint matrices. With this technique, imprints of artificial or natural moiacules including their recognition sites. uch as of molecularly imprinted polymens or biological receptors, antibodies or enzymes, can be prepared. In the former case. using the original irnprints as came orttiol ds in a subsequent polymerization step, utilizing preferentially f unctionaily complementary monomers, the formation of impiint materials containing recognition sites complementary in shape and functionality with the original imprint can be obtained but being of different composition. Altemativcly. imprints or images of the artificial or biological species can be obtained directly. The ao formed preparations can be used in a vant variety of applications, e. g. as new diugs, ttiliibitors or new affinity materials.