CI] 7S7551
`
`PATENT AP?LICATION SERIAL NO:
`
`Y;i#lffillXldti'Htl:3
`iuf *pco*P@
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`090 l{$ 12/05/9r 0??97851
`
`PTO-1556
`$/87)
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`101 t'175'00 cK
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`Page 1 of 249
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`ILMN EXHIBIT 1022
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`

`
`1ilililililil||ilililillil
`
`ililflil]tililtililtl
`tililtill
`
`U.S. PATENT APPLICATION
`
`sERIAL NUilEEK
`
`07 /797 ,551
`
`TILINI'
`
`l,AI G
`
`11/ t9/91
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`CLASS
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`530
`
`UIlUUT AKI
`
`IJNII
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`l8rr
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`RICHARD A. H0UGHTEN, SOLANA BEACH, CA; JULI0 H. CUERVO, LA JOLLA' CA;
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`vERTFIED THrS APPLN lS A CIP 0F 07/701,658 05/15/91
`AND A CrP 0F 07/617,023 1t/21/9O
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`Theenclosedutllitypatenl_applicali_o1 lgglqtedlo !r.s..gerigl-No. 071701,658, ifiled r,lay 16,. leel
`u.s. Serial. No. 021617,023 filed Novenber 2I, 1990--_-,
`rovernber 19, 1991
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`Fce
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`OR
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`OR
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`For:
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`Page 3 of 249
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`/lfnn -/// fl.
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`Express MaiI Ling Label *o. n"rnJr/rrrw
`
`-1-
`
`,-, /
`29:
`qlgruFFrs er _Egumolr4 ltuLIT pLJl. g!:ggIP3
`ttIXTUREg, E8PECIAL,III Of OLIGTOPEPTIDE I,IIXTURES
`
`07 737551
`
`Description
`Cross-Reference to Related ApBlication
`plicptio
`This is a continuation-in-p3*-2)f
`Serial No. 07/7OL,658 fiLed May 16, 1991r\tha
`aEa
`continuation-in-part of application Serial No.
`-
`o7/6L7,o23, flledNovernber r,
`ffiffiI"
`t\
`are incorporated by reference.
`
`Technical Fie1d
`The present invention relates to the organic
`synthesis of oligoneric sequences of compounds. More
`particularly it relates to stepwise synthesis of
`nultiple independent sequences, especially oligoneric
`peptide chains.
`
`Background and Related Art
`Over the last several years, developments in
`peptide synthesis technology have resulted in autornated
`synthesis of peptides acconplished through the use cf
`soli.d phase synthesis methods. The solid phase
`synthesis chenis'r-ry that made this technoLogy possible
`was first described in Merrifield et al. J. Aner. Chem.
`$9g-, 95r2L49-2154 (1963). The rrMerrifield methodrr has
`for the rnost part renained unchanged and is used in
`nearJ.y all automated peptide synthesizets available
`today.
`
`In brief, the Merrifield rnethod Lnvolves
`synthesis of a peptide chain on solid support resin
`particles. These particles typically consist of
`polystyrene cross-llnked wlth dlvinyl benzene to form
`porous beads which are insoluble in both water and
`various organic solvents used in the synthesis p:otocol.
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`The resin particleE contain a fixed amount of amino- oE
`hydroxylnethyL aronatic rnoiety which aerveE as the
`linkage polnt f,or the firEt amino acld in the peptide.
`Attachnent of the first anino acid entails
`chernically reacting its carboxyl-ternrinal (C-terminal)
`end with derivatized resin to for:m the carboxyl-terminal
`end of the oligopeptide. The alpha-anino end of the
`anino acid is typically blocked wlth a !-butoxy-carbonyl
`group (t-Boc) or with a 9-fluorenylnethyloxycarbonyl
`(F-Moc) group to prevent the amino group which could
`othenrise react from partlcipating in the coupling
`reaction. The side chain groups of the anLno acids, if
`reagtive, are also blocked (or protected) by various
`benzyl-derived protecting groups in the form of ethers,
`thioethers, eEters, and carbanates.
`The next step and subsequent repetitive cycles
`involve deblocking the amino-terninal (N-terminal)
`resin-bound anino acid (or terrnlnal residue of the
`peptide chain) to renove the alpha-anino blocking group,
`followed by chenical additlon (coupling) of the next
`blocked amino acid. This process is repeated for however
`many cycles are necessary to synthesize the entire
`peptide chain of interest. After each of the coupling
`and deblocking steps, the resin-bound peptide is
`thoroughly washed to renove any residual reactants
`before proceeding to the next. The solid support
`particles facilitate renoval of reagents at any given
`step as the resin and resin-bound peptide can be readily
`filtered and washed while belng held in a colunn or
`device with porous openings.
`Synthesized peptldes are released from the
`resin by acid catalysis (typlcalJ.y with hydrofluoric
`acid or trifluoroacetic acid), whlch cleaves the peptide
`fron the resin leaving an anide or carboxyl. group on its
`C-terurinal ami.no acid. Acidolytic cLeavage also serves
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`to remove the protecting groupE from the side chains of
`the anino acids in the synthesized peptide. Finished
`peptides can then be purified by any one of a variety of
`chromatography nethods.
`Though most, peptides are syntheEized with the
`above described procedure uEing autourated instruments, a
`recent advance in the solid phase nethod by R.A.
`Houghten allows for synthesiE of nultiple independent
`peptides sinultaneously through uranually perfotmed
`neans. The ttSimultaneous Multiple Peptide Synthesisrl
`(nS!!PStt) process is described in U.S. Patent No.
`4,63L,2LL (1985); Houghten, @L,
`' 92;5131-5135 (1985); Houghten et aI., Int. J. Peptide
`Protein Res. , 4-167.3-678 (1986); Houghten et aI.,
`Biotechnicrues, {, 6, 522-528 (1986), and Houghten' U.S.
`Patent No. 41531 r2LL, whose disclosures are incorporated
`by reference.
`Illustratlvely,
`the SMPS process enploys
`porouE containers such as plastic bags to hold the solid
`support synthesis resin. A Merrifield-type solid-phase
`procedure is carried out with the resin-containing bags
`grouped together approprlately aL'any given step for
`addition of the same, desired amLno acid residue. The
`bags are then washed, separated and regrouped for
`addition of subsequent sane or different amino acid
`residues until peptides of the intended Length and
`sequence have been synthesized on the separate resins
`within each respective bag.
`That nrethod allows nuJ.tiple, but separate,
`peptides to be synthesized at one tine, since the
`peptide-linked resins are maintained in their separate
`bags throughout the process. The SMPS nethod has been
`used to synthesize as many as 200 separate peptides by a
`single technician in as little asi two weeks, a qate
`
`
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`-4-
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`vastly exceeding the output of nost autonated pept,lde
`synthesizers.
`A robotic device for autonated nultiple
`peptide synthesls has been recently commercialized. The
`device performs the sequentlal steps of nultiple,
`separate solld phase peptlde synthesis through iterative
`mechanical-intensive neans. This instrunent can
`synthesize up to 96 separate peptides at one tine, but
`is linited at present by the quantity of its peptide
`yield.
`
`Several research groups have reported the
`synthesis of synthetic conbinatorial libraries of
`peptides. Those reports are discussed below.
`Of interest is work by Geysen et aI., which
`deals with nethods for syntheslzing peptides with
`spebific sequences of amino aclds and then using those
`pept,ides to identify reactionE with various receptors.
`Geysen et aI. rs lrork presupposes that one has a prior
`knowledge of the general nature of the sequences
`required for the particular receptors, so that the
`appropriate group of peptides can be synthesized. See
`U.S. Patents Nos. 4,70.8r'i,t71 and 4r833,O92i P.C.T.
`Publications Nos. WO 84103506 and WO 84/03564, Geysen et
`dl. ' Proc. Natl.. Acad. Sci. U.S.A., $!:3e98-4oo2 (1984);
`Geysen et al., Proc. Natl. Acad. Sci. U.S.A. , 82.2L78-L82
`(1985) i celzsen et a1., in Synthetic Peptides as
`Antigens, 130-149 (1985) i Geysen et aI., J. Inmunol.
`Meth., LO2z259-274 (1987); and Schoofs et al.,
`J. Inmunol., !!-9,:611-616 (1988) .
`In published PCT application PcTlAU8s/ooL65
`(WO 86/00991), Geysen describes a method for detetmining
`so-called rrmimotopesrr. A minotope is def ined as a
`catamer (a pollnrer of pieclsely defined sequence formed
`by the condensation of a precise number of snall
`moLecul.es), which in at least one of its conforr,i.tiotr"
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`has a surface region with the equivalent molecule
`topology to the epitope of which it iE a ninic. An
`epitope is defined as the surface of an antigenic
`molecule which is delineated by the area of interaction
`with an antibody nolecule.
`The ninotopes are synthesized on a series of
`solid pollmer (e.9. polyethylene with a coating of
`grafted acrylic acid) rods having a diaureter of about 4
`nn and a length of about 50 nm. A spacer fomed by
`reaction of the e-amino group of l-Boc-J.ysine nethyl
`ester and then t-Boc-alanine was added to the resj.ns,
`followed by removal of the !-Boc group to provide an
`anino group to be used to begin the syntheses.
`A urixture of blocked amino acids containing
`different anounts of each of the blocked twenty anino
`acids to be used was dissolved in dinethyl forurarnide and
`then coupled to the rods. That first coupling was
`repeated three tinres using conventional solid phase
`synthesis techniques. twenty anino acid residues hrere
`individually next added so that twenty s-mer sequences
`vrere prepared, each having a single, known anino acid
`residue at the anino-terrrinus and a mixture of arnino
`acid residues at each'of the four other positions of the
`chain. Each of those twenty rod-linked peptides was
`then individually reacted with each of the twenty aurino
`acid residues to for:n 400 (2O x 2Ol 6-mer peptides
`having the two anino-terninal positions defined and the
`four reuraining positions as mixtures. Ttro more
`positions of nixtures of amino acids were then added,
`and the terminal anine acetylated to fom N-acetyl
`8-r€rs Linked to the rods those first two arnino acid
`positions were undefined (nixtures), followed by two
`aefined positions, followed by four undefined positions'
`(nixtures), followed by the spacer and then the
`supporting rods.
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`The 400 rod-linked N-acetyl 8-rner peptide
`nixture preparations were then screened in an ELISA
`assay using a nonocLonal antibody to a desired antigenic
`protein. The 8-ners having the best binding to the
`antibody were identifled. Tlso sets of further 8-mers
`that contained the identified best-binding 2-mer
`sequences within thoEe 8-ners were prepared.
`A first set contained nixed anino acids at the
`three C-terninal positions, followed toward the
`N-termJ.nus, by a position containing each of the twenty
`anino aclds rnade by twenty separate coupLingsr.the
`identified 2-ner sequences, two further nixtures at the
`, next two positions, and an N-terninal acetyl group. The
`second group contained rnixed anino acids at the four
`C-terminal positions, the identified 2-mer seqluences, a
`position made by separate couplings of each of the
`twenty anino acids, nixed amino acids as the tetminal
`residues and an N-terminal acetyl group.
`Each of those rod-Ilnked N-acetyl 8-mers was
`again screened in an ELISA with the nonoclonal antibody.
`The best binding sequences for each group were
`ident:',fied, and thus 4-mer, best-binding sequences were
`idenLified.
`The above process of separately adding each of
`the anino acids on either side of identified best-
`binding sequences $ras repeated until an optinuur binding
`sequence was identified.
`The above nethod, while elegant, suffers fron
`severaL disadvantages. First, owing to the srnalL size
`of each rod used, relatively snalL arnounts of each
`peptide is produced. Second, each assay is carried out
`usilg the rod*linked peptides, rather than the free
`peptides in solution. Third, even though specific
`anountE of each blocked anino acid are used to prepare
`the 4ixed anino acid residues at the desired positions,
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`-7 -
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`there is no way of aEcertaining that an equimolar amount
`of each residue is truly present at those positions.
`fn addition, Furka et al., (1988, 14th
`fnternational Congress of Biochemistry, Volune 5,
`Abstract FRs013) described the synthesis of nine
`tetrapeptides each of which contained a single residue
`at each of the amino- and carboxy-termini and mixtures
`of three residues at each position therebetween. The
`abstract futher asserts that those authorsr experinents
`indicated that a nixture containing up to 180
`pentapeptides could be easily synthesized in a single
`run. No biological assays were reported.
`Recent reports (Devlin et a1., &,ie!ge,
`24924O4-4OS [1990] and Scott et aI., S@,,
`249t386-39O [1990]) have described the use of
`recornbinant DNA and bacterial expression to create
`highly complex nixtures of peptides. For example, a
`4S-nucleotide base pair stretch of DNA was syntheEized
`in which the individual nucleotide bases were varied to
`contain all four possible nucleotide bases (guanine,
`adenine, cytosine and .thlmidine) at every position in
`the synthesized DNA chain, except at each third position
`(3, 6, g, etc.) which contained only guanine and
`cytosine. The onission of adenine and thlnnidine at
`every third position in the synthesized DNA removed the
`possibility of chain terminator triplet codons ending in
`A or T, such as TAA or TGA.
`Tbe resulting DNA sequence would then code for
`a mixture of 15-ner peptides with all conbinations of
`the 20 naturally occurring anino acids at each position.
`Those investigators fused the 45 synthetic
`nucleotide sequence to a gene coding for the coat
`protein of a simp}e bacterlophage and created a Jarge
`Ilbrary of these bacteriophages. Each menber of the
`Library contained a different 45-ner DNA fusion sequence
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`-8-
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`and therefore each menber of the library resulted in a
`different 15-ner peptide fused to the outer coat protein
`of its correspondlng futly assernbLed bacteriophage
`particle. Screening of the recombinant bacteriophage
`particles in a biochenical asEay allowed the
`investigators to find individual peptide-coat protein
`fusionE (bacteriophages) that nere active in that assay
`by enrichnent, seLection and clonal isolation of the
`enrictred bacteriophages that contained active peptide
`fusions. By determining the DNA sequence of the cloned
`bacteriophages, the investlgators could deduce which
`peptide sequences were active in their asEay.
`That rnethod yielded severaL peptlde sequences
`from a nixture of 107 or nore reconbinant
`bacteriophages. Each of the 15-ner peptides found
`contained the same four-anino-acld sequence somewhere
`within its overall sequence, thereby allegedly
`validating the assay accuracy and methodological
`approach.
`
`The reconbinant DNA nethod is extrenely
`screening large nurnbers of peptides.
`powerful for
`is Liurited in that the peptides must be
`However, it
`fused to a larger protein aE a result of and integral to
`the design of the nethod. The peptide-protein fusions
`(and corresponding bacteriophage particles) are likely
`to be unreactive in nany biochenical, biological and in
`vivo assays where the peptides nust be present in
`solution wlthout steric hindrance or confor"national
`distortion. In addition, the nethod results in an over-
`representation of some sequences of peptides due to the
`inherent redundancy of the genetic code which has
`sevbral codons per amino acid in some cases and only ond
`codon per anino acid in others.
`Still
`further, neither group reported *data as
`being definitive for the deter:nination of optional
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`peptide ligands for strepavidin (Devlin et a1.) ' or for
`the two nonoclonal antibodies raised against
`uryohemorythinin (Snith et a1.). Neither group provided
`a single specific answer conparabl.e to the expected
`sequence.
`
`More recently, Fodor et aI., Sgielgg, 251.767-
`773 (1991), descrl.bed the sotid phase synthesis of
`mixtures of peptides or nucleotideE on glass microscope
`slides treated with aninopropyltriethoxysilane to
`provide anine functional groups. Predetennined amino
`' acids were then coupled to predefined areas of the
`slides by the use of photomasks. The photolabile
`protecting group I'IVOC (nitroveratryIoxycarbonyJ.) was
`used as the anino-terminal protecting group.
`By using irradiation, a photolabile protecting
`group and masking, an array of Lo24 different peptides
`coupled to the sllde was prepared in ten steps"
`Irnmunoreaction with a fluorescent-l-abeled monoclonal
`antibody $ras assayed with eplfluorescence nicroscopy.
`This elegant rnethod is also linited by the
`small amount of peptide or oligonucleotide produced, by
`use of the synthesized peptide or nucleotide affixed to
`the slide, and also by the resolution of the photomasks.
`This rnethod is also leEs useful where the epitope bound
`by the antlbody is unknown because aLl of the possible
`sequences are not prepared.
`The prinary linitation of the above new
`approaches for the circunvention of individual screening
`of nillions of individual peptides by the use of a
`cornbinatoriaL library is the inability of the peptides
`generated in those systens to interact in a trnormaltr
`manner with acceptor sLtes, analogous to natura\
`interaction processeE (i.e., in solution at a
`concentration relevant to the receptors, antibody
`binding sites, enzlme bindlng pockets, or the like being
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`studied without the exclusion of a large percentage of
`the possible conbinatorial library). Secondarily, the
`expression vector systens do not readily permit the
`incorporation of the D-forns of the natural anino acids
`or the wide variety of unnatural arnine acids which would
`be of interest in the study or development of such
`interactions.
`The interest in obtainlng biological-ly active
`peptides for pharuraceutical, diagnostic and other uses
`would make desirable a procedure designed to find a
`nixture of peptides or a single peptide within a mixture
`with optirnal actlvity for a target application.
`Screening nrixtures of peptides enables the researcher to
`greatly sinrplify the search for useful therapeutic or
`diagnostic peptide compounds. MixtureE containing
`hundreds of thousands or more peptides should be readily
`screened since many biochenical,, biological and sma}I
`aninal assays are sensitive enough to detect activity of
`conpounds that have been diluted down to the nanogram or
`ranqte, the concentration
`even picogran per milliliter
`range at which naturally occurring biological signals
`such as peptides and proteins operate.
`A1most aLl of the broad diversity of
`biologically relevant ligand-receptor (or affector-
`acceptor) interactions occur in the presence of a
`complex milieu of other substances (i.e., proteins make
`up approxinately 5-1o percent of plasma, e.g. aLbunin
`l-3 percent, antibodies 2-5 percent-salts' Iipids/f'ats,
`etc.1 . This is true for virtually all biologicall.y
`active compounds slnce nost are conmonly present, and
`active, at nanomolar and lower concentrations. These
`conipounds are also, in most instances, produced distant"
`fron their affection sites. That a small peptide (or
`other molecuLe) can readily xfindtr an acceptor qystem,
`bind to it, and affect a necessary biological function
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`prior to being cleared from the circulation or degraded
`suggested that a single specific peptide sequence can be
`present in a very wide diversity, and concentration, of
`other individual peptides and stiil. be recognized by its
`particular acceptor system (antibody, cellular receptor'
`If one could devlse a means to prepare and
`etc.).
`screen a synthetic conbinatorial library of peptldes,
`then the notmal exquisite selectivity of biological
`aftectot/acceptor systems could be used to screen
`through vast nurnbers of synthetic oligopeptide3.
`The availability of a wide variety of clearly
`identified peptides ln relatively linited rnixtures would
`greatly facilitate the search for the optirnun peptide
`for any particular therapeutic end use appllcation. At
`the present tine, researchers are hanpered by the
`inability to rapidly create, identify and screen large
`numbers of peptides with specific receptors. Work such
`as reported by Geysen has been valuable where the
`general nature of the required amino acid residue
`sequence couLd be previously determined, so that the
`specific peptldes of interest could be individually
`for:mulated. However, such techniques cannot insure that
`the optinun peptides,are identified for testing.
`It would therefore be of considerable interest
`to have a nethod for the precise synthesis of nixtures
`of peptides in which lndividual peptide sequences can be
`specificalty defined, such that a comprehensive array of
`peptides is available to researchers for the
`identification of one or more of the optimum peptides
`for reaction with receptors of interest, fron which one
`can derLve optinun therapeutic naterlals for treatnent
`of various organisrn dysfunctions. It would also be of
`value for such a process to have the capability to
`produce eErivalent sequences of other types of \
`oligomeric conpotinds.
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`Brief Surnrnary of the Invention
`In one asPect, the invention herein
`contemplates a process that provides for the synthesis
`of conplex nixtures of step-growth oligomers, especially
`peptides, wherein each position in the oligorneric
`sequence chain contains an equinolar representation of
`reacted bifunctional monomeric repeating unit conpound,
`such as an amino acid residue, added at that step. In
`peptide synthesis, the urethod circunvents the problen of
`unequal reaction yiel,ds during addition of blocked amino
`acids reacted as a rnixture in a coupling step in the
`Merrifield solid phase synthesis procedure. Use of the
`present rnethod also provides a relatively rnuch larger
`amount of coupled oligomer than previously contemplated.
`In its preferred enbodiment, the invention
`conternplates the organic synthesis of conplex equinolar
`nixtures of oligopeptide sequences on a solid support
`rnaterial. The equinolar ollgopeptide sequences consist
`essentiall.y of chains of amino acid residues linked
`end-to-end by peptide bonds wherein the anino acid
`residue incorporated at any one position in the chaLn
`can be varied, such as to contain aLl or a conbination
`of the twenty naturally occurring anino acids and/or
`their derivatives. The invention enables synthesis of
`these peptide mixtures with equal and precise
`representation of any anino acid residues at any
`position in the chain at which a mixture of anino acid
`residues is j.ntended to be represented. The process can
`use any type of peptide addition chenistry and
`protocols, but preferably uses the Merrifield soLid
`phase synthesis procedure in protocols sinrllar to that
`of the Houghten SMPS Process.
`In yet another aspect, the invention comprises
`a rnethod for tire identification of one or more optirnurn
`peptides for reaction with a designated acceptof, such
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`that design of therapeutic naterials for treatnent of
`organism dysfunctions involving such receptor can be
`facilitated.
`In its broadest fotm, a process of this
`inventLon is defined aE a process for the synthesis of a
`complex mixture pool of solid support-coupled mononeric
`repeating unit cornpounds, wherein the nixture pool
`containE a substantially equlurolar representation of the
`reacted nonomeric repeating unit compound, such as amino
`acid residues, coupled at that step. In accordance with
`this method,
`(a) a plurality of Eo1id supports is
`provided, each solid support comprised of a particLe
`linked to reactive functionaL groups. The functional
`groups of the solid support react with a functional
`group of each of the monomeric repeating unit compounds
`to be reacted. In a preferred enbodinent, each of the
`solid supports is within a poroug container, the soLid
`support is of a size that is larger than the pores of
`the container, and both.the container and sol-id support
`are substantially insoluble in a liguid nediun used
`during the stePwise sYnthesis.
`'
`(b) A plurality of liquid media is provided'
`each medium containing a different monomeric repeating
`unit compound from a pJ.urality of rnononeric repeating
`unit compounds frorn which the oligomers are to be
`formed. Each of the nononeric repeating unit compounds
`has a first reactive functional group that reacts with
`the reactive functional group of the solid support and a
`second reactive functiona.l group that is capable of
`reacting during the reaction of the solid support
`functional group and the first reactive functional
`group, but is protected fron so reacting by a
`selectively removable, covalently Linked protecting
`group.
`,.
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`-14-
`(c) Each of the solid supports is placed in a
`different one of the liErid nedia and the reactive
`functional group of each solid support is therein
`reacted with a first reactive functional group of a
`monomeric repeating unit compound in that respective
`nedium to couple that rnonomeric repeating unit compound
`to the solid support.
`(d) Eash of the reactions iE nraintained for a
`tfune period and under conditions sufficient for all of
`the reactive functional groups of the solid support to
`coupl_e to the monomeric repeating unit compound to form
`a plurality of rnonorneric repeating uni.t-coupled solld
`supports.
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`(e) Each monomeric repeating unit-coupled
`solid support is renoved fron its respective liquid
`medium, and equimolar amounts of each of the monomeric
`repeating unit-coupled solid eupports are adnixed to
`fotm a reaction product pooL, wherein equal weights of
`the formed pool contain the sane nurnber of moLes of each
`20 mononeric repeating unit-coupled solid support.
`The above nixture pool is useful in a stepwise
`synthesis f,or preparing a complex niv:tu.'e of solid
`support-coupled oligoners wherein one or more positions
`of each oligoner of the rnixture contalnE an equinolar
`representation of reacted monomeric repeating unit
`compound coupled at each synthesis step. If desired,
`the pool forned in step (e) can be used for further
`steps (1)-(o) as discussed in regard to synthesi-s of an
`oligopeptide hereinafter. However, in usual practice,
`steps (f)-(k) are utilized, as discussed below.
`(f) The reaction product pool is separated
`into a nurnber of aliquots of egual weight. Each of the
`aliquots is enclosed in another porous containeq, where
`such preferred containers are used.
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`(g) The protecting groups are selectively
`removed fron the second reactive functional groups of
`the pool to forn a reacted solid support pool having
`free reactive functional groups. ThiE step is
`preferably carried out after the pool, is formed into
`aliquots and those aliquots are re-encLosed, but can be
`carried out prLor to forming the aliquots' and re-
`enclosure is not used where porous containers are not
`used.
`
`(h) Each of the aliquots having free reactive
`functional groupE is placed into one of a nunber of
`J.iquid rnedia, each urediurn containing a different
`rnonomeric repeating unit conpound from a plurality of
`monomeric repeating unit compounds from which the
`oligonrers are to be forted to form a reaction mixture,
`whereLn each of the rnononeric repeatlng unit compounds
`has a first reactive functlonal. group that reacts in the
`reaction mixture with the free reactive groups of the
`aliquot and a second reactive functional group that is
`capable of reacting during the reastion of the free
`reactive functional groups of the aLiquot, but is
`protected fron Eo reacting by .i selectively removable,
`covalently linked protecting {ioup.
`(i) Each of the reactions is naintained for a
`time period and under conditions Eufficient for aII of
`the free reactive functional groupE of the aliquots to
`react with and couple to the respective nononeric
`repeating unlt compounds to form a nurnber of solid
`support-coupled repeating unit reaction products.
`(j) Each of the solid support-coupled
`repeating unit reactlon.products for:rred is removed, and'
`equinolar anount,s of each of thoEe reaction products are
`adnrixed to fonn a reaction product pooJ.. Egual yeights
`of the reaction product pool contain the same number of
`noles of each reaction product.
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`(k) Thereafter, steps (f) through (j) are
`serially.repeated zero or more tirnes until a plurality
`of solid support-coupled reaction products havlng the
`desired nunber of monomeric repeating units is
`synthesized.
`The resulting complex nLxture of oligomers
`contains an equinolar nixture of the plurality of
`monomeric repeatlng unit conpounds at every
`predetenrined position in the chain. The eguirnolarity
`is only lirnited by the accuracy in driving the reactions
`to conpletion and weighing errors in separatlng the
`substantially homogeneously urixed resins into equal
`aliquots.
`
`fn a preferred form, the process of-this
`invention is defined as a ppocess for the synthesis of a
`conplex nixture pool of solid support-coupled amino acid
`residues wherein the rnixture contains an equinolar
`representation of the anino acid residues coupled.
`Here, the preferred enbodiment of u