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WO 2014/025685 A2 13 February 2014 (13.02.2014) P O P C T

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WO 2014/025685 A2 13 February 2014 (13.02.2014) P O P C T (12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property Organization International Bureau (10) International Publication Number (43) International Publication Date WO 2014/025685 A2 13 February 2014 (13.02.2014) P O P C T (51) International Patent Classification: (81) Designated States (unless otherwise indicated, for every C12Q 1/56 (2006.01) kind of national protection available): AE, AG, AL, AM, AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, (21) International Application Number: BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DK, DM, PCT/US2013/053612 DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, (22) International Filing Date: HN, HR, HU, ID, IL, IN, IS, JP, KE, KG, KN, KP, KR, 5 August 2013 (05.08.2013) KZ, LA, LC, LK, LR, LS, LT, LU, LY, MA, MD, ME, MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, (25) Filing Language: English OM, PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, (26) Publication Language: English SC, SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, (30) Priority Data: zw. 61/680,1 11 6 August 2012 (06.08.2012) US 61/681,485 9 August 2012 (09.08.2012) US (84) Designated States (unless otherwise indicated, for every PCT/S2200 1/0/0494 5 July 2013 (05.07.2013) US kind of regional protection available): ARIPO (BW, GH, GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, SZ, TZ, (71) Applicant: JNC CORPORATION [JP/JP]; 2-1, Otemachi UG, ZM, ZW), Eurasian (AM, AZ, BY, KG, KZ, RU, TJ, 2-Chome, Chiyoda-ku, Chiyoda-ku, Tokyo, 100-8105 (JP). TM), European (AL, AT, BE, BG, CH, CY, CZ, DE, DK, ΓΓ (72) Inventor; and EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, , LT, LU, LV, MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, SM, (71) Applicant : TROLIO, William M. [US/US]; 155 Gibral tar Road, Horsham, PA 19044-2353 (US). TR), OAPI (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, GW, KM, ML, MR, NE, SN, TD, TG). (74) Agents: LAVENUE, Teresa et al; Hogan Lovells US Published: LLC, 555 Thirteenth Street, NW, Washington, DC 20004 (US). — without international search report and to be republished upon receipt of that report (Rule 48.2(g)) (54) Title: DUAL ANTI-PLATELET MEDICATION/ASPIRIN RESPONSE AND REACTIVITY TEST USING SYNTHETIC COLLAGEN 1 0 —» —» ft 4 ¾— r - - < — — → —m- 0 0 » o FIG.3 (57) Abstract: The present invention provides tests that measures functional platelet aggregation such as by using Light Transmis - sion Aggregometry Assays (LTAAs) or flow cytometry, using synthetic, self-assembling human type I collagen, methods of predict - ing and measuring an individual's platelet anti-platelet medication sensitivity and residual platelet activity status when the individual is on a dual anti-platelet therapy of aspirin and anti-platelet medication and kits useful in the assays and methods. DUAL ANTI-PLATELET MEDICATION/ASPIRIN RESPONSE AND REACTIVITY TEST USING SYNTHETIC COLLAGEN This application claims priority to U.S. Provisional Application 61/680,1 1 , filed on August 6, 2012, and U.S. Provisional Applications 61/681,485 filed on August 9, 2012, which are incorporated herein in its entirety. This application also claims priority to PCT application, PCT/US 13/4941 8, filed on July 5, 2013, which is incorporated herein in its entirety. BACKGROUND OF THE INVENTION The conventional, primary need for an effective assessment of platelet response and reactivity is in the field of cardiology. The public health incidence and burden of heart attack, stroke and related cardiovascular and thrombotic diseases are well known. The medical community has long recommended the use of aspirin i primary care to reduce cardiovascular, stroke and certain other risks. The use of Aspirin in other area such as DVT prophylaxis, oncology, orthopedics and prevention is driving a renewed interest in and use of Aspirin alone or in combination with other drugs. Additionally compliance testing and personalized medicine initiatives increase the unmet medical need for a test which can provide Aspirin presence and response, presence and efficacy of a second antiplatelet drug and the residual reactivity of a patient's platelet reactivity. Aspirin (salicylate based compounds) ingestion or exposure inhibits the COX 1 pathway and modifies COX 2 enzymatic processes, which then precludes all subsequent events necessary for platelet aggregation. Since the ingestion of or exposure to aspirin can inhibit platelet aggregation, it has been given as a therapy to prevent undesired platelet aggregation, which can be a source of many heart attacks, strokes and other thrombotic events. Despite the benefits of aspirin therapy in many individuals, aspirin therapy is not effective enough in some individuals as it does not cause the desired inhibition of platelet aggregation or its effect is shorter than the dosing interval (some patients may only get 6 to 12 hours of protection rather than 24 hours resulting in an above baseline risk for the patient in the time between doses). In these individuals the residual platelet reactivity is high and the patient's risk is not mitigated. In other individuals, aspirin therapy can be harmful as it creates an increased risk of unwanted bleeding complications because the aspirin seems to block platelet activity altogether so that the blood does not clot when physiologically necessary. Recently, two elements of Aspirin's pharmacodynamic behavior have entered the clinical mindset: Aspirin must be taken at the same time every day to maintain its antiplatelet effectiveness; and the thrombotic risk from failure to keep this schedule is greater than the patient's baseline risk. Thus, physicians often prescribe a course of treatment involving both low dose aspirin and an anti-platelet medication to inhibit unwanted platelet aggregation. This is often called a "dual therapy." Not all patients will respond to the dual therapy or the individual anti-platelet medication in the same way. Selecting and monitoring Aspirin in dual treatment protocols is also necessary because certain antiplatelet drugs like ticagrelor lose their effectiveness if the Aspirin dose is greater than 100 g. There currently is not an effective way to measure a patient's response to dual anti-platelet medication therapy or to determine the patient's residual platelet reactivity. Thus, there is an unmet need for a reliable tool to assess and manage the anti¬ platelet medication response and reactivity on platelet aggregation when a patient is on a dual anti-platelet medication therapy, as well as checking patient compliance with the treatment regimen. Traditionally, a patient's response to aspirin and other anti-platelet medication therapy is assessed by testing platelet activity using a series of platelet aggregation test. The "gold standard" of platelet aggregation tests, light transmission aggregometry (LTA), utilizes collagen from biological sources as the agonist to bring about platelet aggregation, as a measure of the degree or extent of platelet response or inhibition to aggregation. However, there are multiple issues as well as the risk of infectious disease transmission when using biological material. Biologically derived products, whether 'natural,' processed, manufactured by fermentation, cell culture or similar processes, or recombinant, all share the following drawbacks: carry a risk of infectious disease transmission; have lot to lot variability (regarding the ratio of active materials, performance, chemical characteristics, solubility, stability, moisture content, and process contaminants); differing bio-profiles depending upon the location the product was made; differences caused by processing; and environment, geographic and dietary differences affecting the source animal or culture. Biological collagens cannot be diluted to enhance sensitivity or to generate dilution profiles, both of which easily performed with synthetic collagen. It is widely believed that anti-platelet therapy contributes to the reduction of major atherothrombotic complications in cardiovascular, neurovascular and other diseases. In the treatment of percutaneous coronary intervention and acute coronary syndromes, dual anti-platelet therapy when performed at optimal dosing and timing has significantly lowered the risk of thrombotic complications and contributed to positive outcomes. However, an important clinical problem of increased incidence of major adverse clinical events ("MACE") and confounding differences in patient outcomes relates to the variability in patient response to anti-platelet treatments, especially in a dual antiplatelet therapy (a combination of Aspirin and a second agent such as clopidogrel or ticagrelor). Understanding the mechanisms underlying this phenomenon is important to individualizing and improving patient care, long term (maintenance -sometimes referred to as chronic therapy in the literature) therapy and consistent (positive) outcomes. However, a clear and reliable predictive model for responsiveness to anti-platelet therapy is currently not available. Attempts have been made to characterize the efficacy of anti-platelet therapy using platelet function testing but based on current information, and dependence on biological agonists, its routine use is not recommended particularly as costs, complexities and cost effectiveness have not been established, and lack of correlation, standardization and agreement between laboratory methods, and their respective results, is well documented in the literature. Tobias Geislera et al., Circulation 2010; 122:1049-1052. In addition, the inhibitory effects of aspirin on platelets decreases over time in patients on long term or chronic therapy. Violi, F. et al., J Am Cardio, Vol. 43, No. 6, 2004. Further, as evidenced in AstraZeneca's complete prescribing information on ticagrelor (trade name Brilinta® in the US, Brilique® and Possia® in Europe), Aspirin doses greater than 100 mg reduce the ability of ticagrelor to inhibit platelet aggregation. This conflicts with current clinical care guidelines in which increasing doses of Aspirin to Overcome' Aspirin resistance is recommended.
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