co») United States US 2017001571 7A1 c2) Patent Application Publication (o) Pub. No.: US 2017/0015717 Al MOE et al. (43) Pub, Dat Jan. 19, 2017 (64) METHODS AND CompostT10N: COMPRISING TAU OLIGOMERS (71) Applicant: OLIGOMERIX, INC., New York, NY ws) (72) laventors: JAMES G. MOE, Stamioad, CT (US); Eliot J. Davidowsitz, West Hempstead, NY (Us) (21) Appl. Now 18/285,181 - ol Oct. 4, 2016 Related Application Data (68) Continuation of application No. 19/060,143, filed on Sep. 9, 2011, now Pat. No. 9,864,122, filed us appli- cation No, PCT/USO9108776 on Avg. 21, 2009, (60) Provisional application No, 61/189,679, filed on Aug, 20, 2008 Publication Classification (51) Inch COT 1447 AIK 38/17 (200601) (200501) CO7K 14/47 (2013.01); ABIK 38/1709 Q01301) 7 ABSTRACT Tau protein has a causative role in Alzheimer’s disease and ruiple other nenrodegenerative disorders exhibiting tau histopathology collectively eed tavopathies. The primary Function of tau protein is Wo facie assembly and min tenance of microtubules in neuronal axons. In the disease process tau protein becomes modifiod, loses its afinity to ricrotubules and accumulates in the cell body where it forms aggregates. The lage neurofibrillary tangles formed from tau protein assembled into filaments were thought to be the pathological structure of tau, However, more recent work indicates that smaller, soluble oligomeric forms of tau are best associated with neuron loss and memory impairment, Here, novel compositions of tau oligomers and. novel mechanisms for au oligomer nvcleation, extension and termination are taught, Methods for producing and purifying these structures forthe development of small molecule and mmunotherapetes as well as antibodies for biomarkers of renrodegenerative diseases are taught.US 2017/0015717 Al Jan. 19, 2017 Sheet 1 of 25 Patent Application Publication snujue}-9 Nore isnujue}-N Nie 1 e3] Lge Nzrie pager Oly ay Nanay eu by Lpb “L aunbig east |US 2017/0015717 AL Jan. 19,2017 Sheet 2 of 25 Patent Application Publication zzesho 94Hd Lezsko x94Hd . uolBey YO-SUl|Old Lov La pugengzuyru| zd | td urewop Ajquiessy ulewop uoKoelo1dg (NZray) bpp nel ‘Z asnBlyPatent Application Publication Jan. 19, 2017 Sheet 3 of 25 US 2017/0015717 Al Figure 3, HIPPOCAMPUS SECTION Lo SUBICULUM : | ENTORHINAL CORTEXPatent Application Publication Jan. 19, 2017 Sheet 4 of 25 US 2017/0015717 Al Figure 4. tau oligomers Pathological )US 2017/0015717 AL Jan. 19,2017 Sheet 5 of 25 Patent Application Publication yjyeaq uosneN Ro ° saawobijo ne} aeinyjeoesyxy uoniqiyut onedeseyjounwuy “g ounblg uojnen AuyeeH { siOyIEWOIG siewobijo ne} aejn}jaoe4}U]Patent Application Publication Jan. 19, 2017 Sheet 6 of 25 US 2017/0015717 Al Figure 6. Tau Oligomer (RFU/mi CSF) 09! Severe AD AD non-ADPatent Application Publication Jan. 19, 2017 Sheet 7 of 25 US 2017/0015717 Al Figure 7. Non-Reducing Reducing Conditions = Conditions 3 ae aa F423 45 Disulfide- Linked tau oligomers Monomer J} Tau sctrms Tau degradation productsPatent Application Publication Jan. 19, 2017 Sheet 8 of 25. US 2017/0015717 Al Figure 8. Heat and Reductant 0) 4) a F123 45 5 Trimer — Dimer — Monomer —Patent Application Publication Jan. 19, 2017 Sheet 9 of 25 US 2017/0015717 Al | C-terminal tau,,, oligomers C-terminal tau er = Figure 9. I a g 3 & E £ zPatent Application Publication Jan. 19, 2017 Sheet 10 of 25 US 2017/0015717 Al Figure 10. Hexamer 6° Pentamer (5°) Tetramer (4°) Trimer (3°) Dimer (2°) Monomer (1°)Patent Application Publication Jan. 19, 2017 Sheet 11 of 25. US 2017/0015717 Al Figure 11. 2US 2017/0015717 AL Jan. 19, 2017 Sheet 12 of 25 Patent Application Publication ~— D006 — mo EEA: Oeli0z'c0V seul, peyuind | Se se me 06 {oz 108 208 0 109 fos Rear Jew paylind “@L eunbi4Patent Application Publication Jan. 19, 2017 Sheet 13 of 25 US 2017/0015717 Al Trimer Figure 13. Dimer min. 60-min.. 15.min. 60 min.Patent Application Publication Jan. 19, 2017 Sheet 14 of 25. US 2017/0015717 Al D ne ee ee Figure 14. | i | M suawobijo ney JappeyUS 2017/0015717 AL Jan. 19, 2017 Sheet 15 of 25 Patent Application Publication (yp) sauoUOW | pyne| —-+| *YEWOBIO + (ZOZH) UOHEPIXO, BN ON + + uoeqnoul “gy ounbiUS 2017/0015717 Al Jan. 19, 2017 Sheet 16 of 25 Patent Application Publication duuuddduuduuddddududddududddduddduuvuuddudduduudddudddududdddudu SAAGSMAA 1S YOHGLNWAWNG UAT TAHLEI MAND DOG AULINGTSSINSOAUCNEOTHESHARA eR te + =F teh ee (s+ Trezeao ‘ee peBxeyo eT) Mf (T+ TTezeac ‘ee peSzeyo TT) pe duuuuddduduuduudddudduduudduududuuuuuddudddduuddduddududdduududuuwudddudddduudddudduduuuuuudd ODOHANHH INDTSHON SLANS TAAAMAALOASDOOAAHM I NOASDONSOANS TATHANT LOANS GOWN INGLSOINSMANNTAANSASVIO +t + + = + tonto ~ +t + to th EH (vt Trezea0 ‘ee pebzeyo 1) EY (b+ TTeeAo ‘ee peBxeyo g) ZY (+ TTeZeAo ‘ee poBzeyo a) TE suyeuog Surpute eTAgn;oxoTH AEE Ue “QL eunBi4Patent Application Publication Jan. 19, 2017 Sheet 17 of 25 US 2017/0015717 Al ao Qo ao ¥ O-— ow is a 7 + 2 O—o ® i § o-oo [4 ¥ o-o 3RPatent Application Pul blication Jan. 19, 2017 Sheet 18 of 25 US 2017/0015717 Al Figure 18. BS Oe R3-3R R3 Besheet regions =)US 2017/0015717 Al Jan. 19, 2017 Sheet 19 of 25 Patent Application Publication 2v 8s [ge ME] 20°97 e6'sv UP! es69 [6L were levee [eres _UPraE| lsros ‘sete Ubrab} StL eS6hh MEMbYE ee Zeb 9g'8z) UPA SE ZS SEL ieZ Zeb Mbrebreb LzosL eysoL _|Seubruvee| \es OLE 6S rdt PSP PISE ez'eeh zzeeh Ubrebbryy MN mw quereddy| _ payeinsjeo) JeWOUoW Jewig sown we/prprds ——___ JOWeIOL { Uprivriyras — > ‘UbRsbhbite ————— “6b ounBi4Patent Application Publication Jan. 19, 2017 Sheet 20 of 25 US 2017/0015717 Al Figure 20. ARRS ARR? \ ih & \ oe! & RR RRS Se R2-3R RB B-shest regionsPatent Application Publication Jan. 19, 2017 Sheet 21 of 25 US 2017/0015717 Al Figure 21, wn Det be ARR3 | => cy 4R RO-3R R3 B-sheet regions AR R3-3R R3 B.sheet regionsPatent Application Publication Jan. 19, 2017 Sheet 22 of 25 US 2017/0015717 Al Figure 22, R R3-3R R3 f-sheet regionsPatent Application Publication Jan. 19, 2017 Sheet 23 of 25 US 2017/0015717 Al Figure 23. 4R R2-3R R3 Bssheet regions AR R3-3R Ro B-sheet regionsUS 2017/0015717 Al Jan. 19, 2017 Sheet 24 of 25 Patent Application Publication spToe OUTWe 62 HOOD-HHINOTISOONSLANSTOAGMAATOASDOO-NZH z epradea spToe outue 0€ HOOD-AHNINGNSDONSOANS TOTMANIIOAMSOD-NZH T eptadea ‘ve einBi4US 2017/0015717 Al Jan. 19, 2017 Sheet 25 of 25 Patent Application Publication puog epuinsiq uojBeu eoys-9 Ex YE-TY UP SS ooE San SPSS GAANXAIONS95 AHMINGMS SOANSTOTINNITOAMSSS TLée “Gz eunBi4US 2017/0015717 Al METHODS AND COMPOSITIONS COMPRISING TAU OLIGOMERS 0001] This application claims the benefit ofthe fing date ‘of US, Provisional Application No, 61/189,679, filed Aug. 20, 2008, entitled “Methods And Composition Comprising ‘Tau Oligomers.” The entie disclosure is hereby incorpo- rated by reference into the present disclosure BACKGROUND [0002] Alzheimer’s disease (AD) is the most common ans of dementia inthe elderly ht ffs an estimated 15 milion people worlwide nd 40% ofthe population above 85 yeas. Te cists is harcterizd by progressive loss of msm, psec end movement with total inepactaton of the pict and eventually death AD tas a tel all on thowe wth the disco well thr fils, Sends and ceaegiver. [0003] ‘The symptoms of AD manifest slowly andthe fest Symptom may only be mild fngtnes, In this stage, individuals may forget recent events, activities, the names of familar people or things and may not beable slve simple math problems. As the disease progresses into moderate Stages of AD, symptoms ae mor tsi noice an become Serious enovgh 10 case people with AD or their family members sek metal help. Moderate-stage symptoms of AD incude forgetting how to do simple tsks such as {smoming nd problons develop with speaking. undrstand- ing, reading, oF writing. Severe sage AD patents may hocome anxious or aggressive, may wander aay’ fo home and ultimately need ttl cae [0004] No core is curently aaiable for AD. Tey: inedcaion therapy focuses on controling the symptoms of AD and is varius stages, Fr example, mild vo moderate AD can involve tetent with cholinesterse inhibitors such as Cognex® (tacrine), Aricept® (donepezil), Exelon® (vastiamine), or Razadyae (galantamine). Wheres mod rato severe AD canbe treated with Namen (memn- tng). These medications may help delay or peveat AD Symptoms fom becoming worse fora limited period of tine So eal AD treatments warned. However thee i no cea evidece that these medications have any eat on the underying progression ofthe disease 10005] ‘There i a large and rapidly growing unmet need for disease modifying drugs for Alzheimer’s disease (AD). The classical hallmarks of AD are intrneuroal plaques consisting of povpttes or aggregnes of alo beta protein (Ap), and intra-neuronal neurofibrillary tangles (NET) of ta protin. Tau protein prometes microtubule assembly and sail and serie fr the faneton of jxons,whsres the nomual funtion of AB is not filly understood. The amyloid cascade hypothesis has been ‘viel accepted a the pathological pathway of AD. Teholds that the generation of AB and accumulation of AB aggregates inthe brain iit th disease process. I is supped by senetic evidence that mutations leading increased acu mulation of AB aggregates leads to familial AD, However, ther ars umber of wenknescs inthe A cascode hypoth cis in that it doesnot adress the importance of ter pathways that can cause neurodegenerstion (Seabrook el 2007). The acumlation and distbaton of NFTs in the brsins of AD patients is highly corelated with disease progression and can be tsa fo age AD by postmortem Bruin hstopabology, whereas tha is poor conelation Jan. 19, 2017 between AD and the accumulation of neue plagues com- posed of beta amyloid. This has been used to challenge the ‘anyloid hypothesis Josephs et a, 2008), Lockluster results for AB directed therapeutics in lat stage clinical trials has increased interest in exploring allemative targets for drug iscovery such as tau (Iabal etal, 2008), [0006] While extensive research in the past decade has ‘denied possible biomarkers foe AD, there i stil an urgent reed for composition and methods that are specifically usefl in diagnosing, stratifying, oF monitoring the progres- sion oF regression of AD. Ness compositions and methods fare also needed that serve as dag targets for the identifica- tion of new medication therapies to teat AD and to monitor illereat medications therapeutic ellect when used to teat AD, as well as compositions that are usefil as immuno- therapeutic agents SUMMARY [0007] The present application provides methods and compositions comprising tau oligomer of a fragment oF peptide derivative thereof. These tau oligomers, fragments ‘or peptide derivatives thereo can be used in diagnostic and prognostic assays, allowing AD to be diagnosed earlier (bile the patient is alive) and more accurately than was previously possible. ‘These tau oligomers, fragments or peptide derivatives thereof can better help the clinician stratify, or monitor the progression or regression of AD, than currently available assays. In addition, tau oligomers, frag- ments oF peptide derivatives thereof identified acconding to the composition and methods disclosed ean serve as dru targets forthe identification of new therapeutic agents forthe treatment of AD or other tauopathies and monitor diferent medication therapies benefit when used to teat AD or other tauopathies. In some embodiments, these tau oligomers, fragments or peptide derivatives thereof can be used a Jimmunotherapeutic agents to stimulate the immune system, {0008} In some embodiments, there isa novel model for ‘au oligomerization that provides novel compositions of tau oligomers that help teach how the relative overexpression of 4 tau can facilitate tan aggregation and neurodegenerative disease and how these strictures may be used for discovery of small molecule drugs, immunotherapedtics and biomark- es for neurodezeneratve diseases. {0009} In one embodiment, composition comprising Stabilized ta oligomer or fragment or peptide derivative thereof, [0010] In another embodiment, a composition is provided, ‘wherein (the ta oligomer isin atleast one conformation comprising tau dimer, ta trimer, tau tetamer, tau pentamer, ‘au hexamer, tau septamer, tau octamer, tau nonamer, tat decamer, tau uadecamer, tau dodecamer,3R tau, 4R tau, oF mixtures of 3R tau and 4R tau of a fragment or peptide derivative thereof or a combination thereo! or (i) the tau ‘monomeric unit comprises one of SEQ ID NO. 1-6: o¢ (ii) the ta oligomers stable for at east 2 months; or Gv) the tau oligomer is substantially purified and/or isolate. [0011] In some embodiments, a method is provided for screening an agent for modulation or disruption of purified ‘au dimer, tu time, ta tetramer, tau pentamer, au hex- amer of a combination thereof, the method comprising: a) contacting a sample containing tau dime, tau trimer, tau tetramer, tau pentamer, tay hexamer of a combination ‘hereof with an agent suspected of being capable of modu Iating tu oligomer formation or disrupting tau oligomers;US 2017/0015717 Al ‘and b) detecting the amount of tau dimer, tw trimer, tau tetramer, tau pentamer, tat hexamer or a combination thereof, wherein deerease in tau dimer, tu trimer, tau tetramer, tau pentamer, ti hexamer or a combination thereof indicates that the agent modulates tau oligomer formation or disrupts tau oligomer [0012] In some embodiments, a composition is provided ‘comprising a tau dimer, which comprises tau 4R-tau 4R. dimer, au 3R-4R tau dimer, or tau 3R-tau 3R dimer. [0013] In some embodiments, a compound is provided that prevents tau-tau binding resulting from hydrogen band- ing, van der Waals interaction such that disulfide bond formation is inhibited or prevented, [0014] In some embodiment, 9 method is provided for feveening an agent for modulation or disruption of tau ‘oligomer, wherein the agent comprises curcumin, demethoxycurcumin, —bisdemethoxycureumin, andlor morin [0015] In some embodiment, «method is provided for Sereening an agent for modulation or disruption of tau ‘oligomer, wherein the ageat disrupts tau-tau oligomeriza- tions by inhibiting tau-t binding, [0016] In some embodiments, atau oligomer composition is provided comprising tau monomer units bound to each ‘other by one or more disulfide bonds [0017] In some embodiments, a tan oligomer composition is provided, wherein he ta oligomer is tu dimer bound to ‘each other by intermolecular disulfide bonds. [0018] In some embodiments, atau oligomer composition is provided, wherein the tau oligomer is tau dimer bound to ‘each other by intermolecular disulfide bonds and the tau dimer has 3 or 4 microtubule binding repeat regions. [0019] In some embodiments, atau oligomer composition is provided comprising one or more fragments of au com- prising R2 R3-R3, oF R3-R3, or R2RI-R2R3 compleres held together by one of more intermolecular disulfide bonds [0020] In some embodiments, a reagent composition is provided comprising a predetermined ratio of tau 3R to tau 4R used 0 generate tau oligomers of from about two to about twelve tau monomer unis. [0021] In some embodiments, a weagent composition is provided used to identify a pathological binding partner associated with AD or tauopatis. [0022] In some embodiments, a reagent composition is provided used to perform high resolution NMR, X-ray ‘cystallography, generating antibodies t specific oligomer sizes, or for immunotherapy. [0023] Additional features and advantages of various ‘embodiments will he set firth in part inthe desription that, follows, and in part will be apparent fom the description, or may be leamed by practice of various embodiments. The objectives and other advantages of various embodiments will be weaized and attained by means ofthe elements and ‘combinations particularly pointed out in the description and ‘appended claims BRIEP DESCRIPTION OF THE DRAWINGS 0024] In part other aspects, Features, benefits and advan- tages ofthe embodiments wll be apparent with regard to the following description, appended claims and accompanying drawings where: 10025] FIG. 1 illustrates the six tau protein isoforms that result from alternate splicing wit the N-terminal inser in the acidic domain (E2 and 3), the repeat domains of the Jan. 19, 2017 microtubule binding region (R1,R2, R3 and Ré) and the tau pseudo repeat (R) indicated. Note tht there are three tau protein isoforms with three repeats (1382, tu381 and tau410 only contain repeats RI, R3 and RA) and three tau protein isoforms with four repeats (@u383, taudl2 and tau441) inthe microtubule binding domain (RI, R2, R3 and Ray, {0026] FIG. 2iustratestau44 structural features inchd- ing the projetion damain (N-terminal) the proline rich region, and assembly domain (C-teminal), Also indicated is the betasheet forming regions (PHE6* and PHEF6) and the two oysteines (Cys291 in R2 and Cys82 in R3).AILAR tau protein joforms contin four repeat regions as shown (RI, 2, RB and 4), whereas 3 tan protein isoforms only contain three repeat regions (R1, R3 and Ré). (0027) FIG. 3 illustrates the contiguous spread of tau pathology is a hallmark of AD. As AD progresses, tay pathology engulfs the hippocampal structure in highly Selective and ondely fashion [0028] FIG, 4 illustrates the proposed mechanism of tau oligomer-madiated disease propagation. An upstream event such as oxidation, physical damage, chemical damage, infection andlor effects mediated by upstream A indvced pathology (A.); leads to accumulation of tau off of micro- tubules and intracelilar ta oligomer formation (a dimes, tau trimer, et.) (B,}, cell death results in release of tau oligomers extocllulary (C.); whereby extracel tau oligomers bind to healthy neurons (D.); leading to the uptake of tam oligomers into healthy cells, and the subsequent promotion of te formation of additional ta oligomers and a diseased cell phenotype (E,); The rate of propagation is directly proportional tothe level of extracellular tau oi- omer (F) [0029] FIG. $ illustrates the therapenic intervention points for halting or aresting disease progression based on small molecule drugs for intracellular and extracellular tu oligomers (mest disease process within & neuron and anest spread of tan pathology to neighboring healthy cells), and {or active or passive immunotherapy targeting extrceliular ‘av oligomers (arest spread of tau pathology’ to neighboring healthy cells). The extracellular tau oligomers are useful biomarkers for disease progression. [0030] FIG. 6 ilustates that ta-oligomer levels in CSF scoumulate progressively during disease as determined by ‘an oligomer conformation-specific ELISA. using capture antibody against tan c-ermimus. The subset of tau oligomers containing tau with an intat eemninus was studied using capture antibody mAb 46, spevific for the 37 carboxyl temninal amino acids of tau, t0 improve the resolution of ‘oa-AD, moderate AD and severe AD samples in combina tion with detection antibody pAb ATL (0031) FIG. 7 ithstrates that dsalfide-tinkes tan oligom- ers are present in the cerebrospinal fuid (CSF) of AD patients illustrating the biological relevance of extracelilar Aisulide inked tan oligomers {0032] FIG. 8 illustrates that tau in CSP is “reactive” in that it contains reactive thiols and forms higher onler auaregates if accelerated by temperature elevation as shown in the le panel. The pane en the right shows the higher onder agasepates that are stable to relucing conltions, SDS, and heat (0033) FG, 9 ilstrtes 4R tau oligomer form driven by the microtubule binding domain inthe C-termi: tus, A tu fragment (18.5 KD) that encompasses the C-te-US 2017/0015717 Al minus is able to oigomerize (charge approximately +13.2) whereas a tan fragment (21.5 KD) that encompasses the Netermius does not form any significant higher order structures (charge approximately -72). Shown in lane | is the MW ladder, lane 2 tau4dl monomer, lane 3 Tans] oligomer mixture, lane 4, the N-terminal portion of tau protein (amino acids 1-184) does not aggregate, and lane S ‘he C-terminal fragment aminoacids 183-481) that contains the full 4R microtubule binding domain, [0034] FIG. 10 illustrates that polymerization of tau oli- omers occurs by addition of monomeric tau to a growing chain resulting in progressively longer tau oligomers as shown for tau]. Evidence for oligomer building via ada- tion of monomer tau based on accumulation of lower order structures followed by higher order strectures as shown, Furthermore, purified monomer is able to form dimers, ‘whereas purified dimer and trimer do not show any higher ‘onder oligomers alier 2 months at 4°. [0035] FIG. 11 illustrates that tu oligomers can be puri- fied in stable form as shown for a mixture of tand12 ‘oligomers that was purified. To prepare specifi tv oligom- ‘ers at high purification Tau? oligomer enziched prepara: tions (lane 2) were size-fractionated to isolate monomer (lane 3), dimer (lane 4), timer (lane S) and tetzamer (Lane 6) ‘and stabilized in butler. [0036] FIG. 12 illustrates that disulfide mediated tau of omers have high thermal stability up t0 90” C. [0037] FIG. 13 illustrates that tau oligomers are highly Stabile under high salt conditions at 37° C. Tavd4] where [L—low salt; M~medinm salt; and H—high salt, [0038] FIG. 14 illustates the temperature stability of ‘auddl oligomers and purified dimer and trimer with and without reducing agent (DTT) Both the dimer and trimer show good stability at 37° C. (incubated at 15, 30 and 60 rinotes) Dimer formation dependent on the formation of ‘one of more disulfide bonds. Trimet formation —demon- strates at least one thiol group is available in the dimer M_purified monomer D is purified dimer: D® is purified «dimer plas reducing agent; T—purifed timer; and T*—pu- ried trimer plus reducing agent. [0039] FIG. 15 ilustrates intrinsic differences in tau iso- form oligomerization that provides the basis forthe disulfide mediated tau oligomer model, 3R tau can only form dimers ‘and has a single eysteine (Cys322); whereas 4R tau forms nl oligomers without temnination andl has two cysteines (Cys291 and Cys322). Oxidative conditions caused rapid ‘oligomer formation due to oxidation of free thiols in mano- erie ta. [0040] FIG. 16 illustrates microuibule binding domain structural features through the repeat and pscudo repeat regions, The amino aid charge i indicated shove the amino acid abbreviation. The beta-sheet forming regions and eys- ‘eines ate highlighted in bold, Potential proline or glycine mediated bending regions are underlined. The microtubule binding regions are defined as follows: RI=Q244 to K274; R2=V375 to S305; R3=V306 to Q336, R4=V337 to N368, ‘and R'=K369 to $400. In 3R tau isoforms, R2 is absent [0041] FIG. 17 illustrates the model for formation of disulfide mediated tau oligomers. For 4R tau protein is0- forms, there are two cysteines and reaction with monomer oF ‘oligomer leaves a free thiol available for reaction, For 38 tau protein soforms, there isa single cysteine which terminates 8 growing oligomer chain. In healthy brains, the ratio of Jan. 19, 2017 SRIAR isoforms is approximately 1.0; whereas in AD Brains the ratio of 3RVAR is approximately 02-03, [0042] FIG. 18 illustrates the model of a disulfide medi- ated strecture for a 3R/3R tau dimer showing alignment of the peptides in parallel to foem a beta sheet structure [0043] FG. 19 illustrates the tu oligomers formed using various ratios of 3R to 4R tan protein isoforms during reaction. The aecumulation of higher order aggregates indi- cates thatthe dimer is reactive consisteat with the formation of only a single disulfide in the initial 4R/4R structure Incorporation of 3R tau limits oligomer exteasion, Higher order oligomers formed with increased levels of 4R tau. The table shows the calculated and appareat moleculae weight forthe 3R/3R 4R/AR and 3R and 4R tau oligomers formed [0044] FG. 20 illustrates the model of a disulfide medi- ated strocture for a 4R/4R tau dimer showing alignment of the peptides in parallel to form a beta sheet. The formation cof an intermolecular disulfide Tinkage traps the tau proteins into an aggregation template that contains wo reactive free thiol groups. [0045] FIG. 21 illustrates the model of a disulfide medi- ated structure for a 4R/R/3R tau trimer showing alignment ‘ofthe peptides in parallel to form a beta sheet, The reaction of 3R tau protein to the 4RV4R tam dimer eliminates occurs via additional formation of a disulfide linkage. This results in the case of the 4R/4R/3R tau timer with a single remaining reactive fee thiol group. At this time, there is no ‘way to prediet whether the disulfide is between 3R 3 anki AR R2 or 4R R3. The trimer is shown with the disulfide linkage to 48 RS strictly for illustrative purposes. [0046] £1G. 22 illustrates the model of a disulfide medi- ated structure for a 3R/4R/4R/3R tau tetramer, showing aligumeat of the peptides in parallel w form an additional beta shoot structe. The 3R/4RI4RI3R tau tetramer con- tains no reactive free thiol groups and cannot propogate isulfide mediated tan oligomers [0047] FIG, 28 illustrates stacked tu tetramer structure (QR/4R/4RV3R that is stabilized by the stacking of the beta sheet regions and by three intermolecular disulfide linkages [0048] FIG. 24 ios examples of two tau peptides that encompass the key structural elements for the tau oligomer dimer that will be used to generate tau oligomer fie monoclonal antibodies. The two peptides were designed to contain all important structural elements leading ‘o dimer formation including the beta-sheet forming regions, cysteines for forming intermolecular disulfide bonds, and the amino acids located between the proline cap regions (Pr0270, Pru8OL, Pro832) that disrupt secondary structure tana or provide for a bead in the protein backbone of the pareat molecule. Additional peptides may be designed with Increased or decreased residues on COO— or amine temni- nus (plus or minus ten amino acids), or mutations leading to ‘amino acid substitutions found in tamopathies. The beta- sheet forming regions and eystenes a highlighted in bold [0049] FIG. 25 illustrates a stable taw peptide dimer anti- en that will be generated to isolate tau oligomer specific ‘monoclonal antibodies. Tau oligomer specific monoclonal antibodies will be used for drug discovery and diagnostic purpose, but also have utility as potential passive immu- notherapeutc agents. The peptide fragments will be adjusted and tested using structural information (Le. circular dichro- jsm and fourier transform infrared spectroscopy) to select tau peptide dimers with beta-sheet structure. Hoth R2+R3