HK1137808B - Use of copertin or neurophysin ii in the manufacture of a kit for risk stratification for acute coronary syndrome - Google Patents

Description

Application of copeptin or posterior lobe hormone carrier protein II in preparation of acute coronary syndrome risk grading kit

Description of the invention

The invention relates to a method for risk stratification of Acute Coronary Syndromes (ACS), in particular Acute Myocardial Infarction (AMI) and Angina Pectoris (AP), wherein the prodiuretic prohormone (provasopressin, proAVP) or fragments and partial peptides thereof, in particular copeptin or neurophysin II, are determined by means of in vitro diagnosis. The invention further relates to suitable combinations of biomarkers for in vitro diagnosis.

Risk stratification (Risk stratification) is becoming increasingly important in symptomatic as well as asymptomatic cardiologies. In particular, for acute coronary syndromes, there is a great need for an appropriate risk stratification.

Risk stratification is used to identify patients with poor prognosis, for intensive care diagnosis and treatment/management, with the aim of being able to achieve optimal clinical outcome. Thus, the goal of appropriate risk stratification is to utilize percutaneous coronary intervention and current agents for effective treatment of acute coronary syndromes.

Appropriate treatment requires early diagnosis and identification of acute coronary syndromes, even after an emergency room admission (emergency room administration), along with clinical decisions. Due to non-specific symptoms in acute coronary syndromes, such as chest pain, differentiation and description from other diseases and identification of acute coronary syndromes are essential.

In the prior art, biochemical markers, in particular classical markers such as cardiac troponin, myoglobin and CK-MB mass, have been used in attempts to predict myocardial infarction (Katus, H.A.; Remppis, A.; Scheffold, T.; Diederich, K.W., and Kuebler, W. (1991): Intracellular compatibility of cardiac peptide T and its release kinetics in tissues with treated and nonperfused myocardial infringement, Am J Cardiol 67 (16): 13601367). B-type natriuretic peptide (BNP) together with pro-BNP (NT-proBNP) (EP 1363128B1, EP 1666881A2) has proven to be another effective biochemical marker in myocardial diagnostics.

And peptoids (also known as C-terminal proAVP) have been described in WO 2006/018315(BRAHMS AG) as biomarkers for the in vitro diagnosis of AMI. Corresponding copeptin analyses are disclosed in Morgethaler et al (Nils G. Morgethaler, Joachim Structure, Christine Alonso, and Andrea Bergmann, Assay for the measurements of copeptin, a stable peptide derivative from the predictor of vasopressin, Clinical Chemistry 52: 112-.

Posterior leaf hormone transporters have been described as nicotine uptake (Robinson A.G., Isolation, assay, and section of induced human neuronopathy, J Clin Invest 1975; 55: 360367), malignant and non-malignant related symptoms of ADH secretion dysfunction Syndrome (SIADH), and markers of renal diabetes insipidus (Pullan P.T., Clappison B.H., Johnson C.I., plasmid vasopression and human neuronopathy in physiological and pathological conditions with change in expression, J Clin Endocanol methyl 1979; 49: 580587; North W.G, Roller F.T., J.J., Melton J.M.M., Mill R.C.A.C.1981; analysis J.894: the family of microorganisms J.1984).

However, the known diagnostic methods using markers known to date have the disadvantage that an early and complete identification of the at-risk patient (at-risk patient) is not achieved, and therefore the risk stratification is not sufficient. It is therefore an object of the present invention to develop a method for risk stratification of acute coronary syndromes which enables an improved identification of at-risk patients.

A further disadvantage is that in the prior art, sufficient sensitivity and/or specificity of the marker is not usually achieved.

It is therefore a further object to provide a method for risk stratification of acute coronary syndromes, wherein at least one marker or a combination of markers has sufficient sensitivity and specificity in an in vitro diagnosis.

It is therefore an object of the present invention to provide a method for risk stratification of acute coronary syndromes. This object is achieved by using a method for risk stratification of acute coronary syndromes, wherein proantidiuretic hormone (proAVP) or fragments and partial peptides, in particular copeptin or metaproten II, are determined by means of in vitro diagnostics (hereinafter referred to as "method according to the invention").

Surprisingly, proantidiuretic hormone (proAVP) or fragments and partial peptides thereof, in particular copeptin or neurophysin II, have a high sensitivity and specificity for diagnosing acute coronary syndromes (cf. examples and figures).

The term "acute coronary syndrome" includes coronary heart diseases in different stages, which are directly life-threatening. This is particularly relevant for emergency medical care, in particular acute myocardial infarction and/or angina pectoris, and sudden cardiac death. In addition to Acute myocardial infarction, which according to the WHO standard (WHO (1979) is Nomencuture and criticism for diagnosis of abnormal coronary disease, Report of the Joint International Society and Federation of diagnosis/World Health Organization for diagnosis 59 (3): 607609), which is defined as an Acute chest pain event lasting more than 20 minutes together with ST elevation and/or myocardial enzyme increase, the term "unstable angina" (AP) has been established which according to the invention is interpreted as "Acute coronary syndrome" (Hamm C. W.: leitline: 2004: Akutes syndrome (ACS) -Tell 1: ACS-patent syndrome [ ST: 2004: heart disease ] recovery 1: tissue syndrome (ST: delivery: 7293; yield: heart disease: 76).

According to the present invention, the term "risk stratification" comprises the identification of patients with poor prognosis, in particular emergency care and at risk patients, for the in-depth diagnosis and treatment/management of acute coronary syndromes, in particular myocardial infarction and angina pectoris, with the aim of being able to achieve an optimal clinical outcome. The risk stratification according to the invention thus allows an effective treatment of acute coronary syndromes with percutaneous coronary intervention and with the latest agents.

Thus, the present invention further relates to the identification of patients with an increased risk and/or poor prognosis of acute coronary syndromes, in particular myocardial infarction angina; and symptomatic and/or asymptomatic patients, particularly emergency care patients.

The method according to the invention can be used in a particularly advantageous manner in emergency and/or intensive care to provide reliable stratification. Thus, the method according to the invention allows clinical decisions, which lead to a rapid treatment success and avoid necessaries. Such clinical decisions also include further treatment with drugs for the treatment or therapy of acute coronary syndromes, particularly Acute Myocardial Infarction (AMI) and Angina Pectoris (AP).

The invention therefore further relates to a method for risk stratification in a patient with acute coronary syndrome for making clinical decisions, e.g. for further treatment and therapy with drugs, preferably in time-critical intensive or emergency medical care.

In a further preferred embodiment, the method according to the invention thus relates to the therapeutic management of acute coronary syndromes, in particular Acute Myocardial Infarction (AMI) and Angina Pectoris (AP).

In a further preferred embodiment of the method according to the invention, a risk stratification is carried out for prognosis, differential diagnostic early detection and identification, severity assessment, and progress assessment of concomitant therapy.

In a further preferred embodiment, the invention relates to an in vitro diagnostic method for the early diagnosis or differential diagnosis or prognosis of myocardial infarction or angina pectoris, in which the marker proantidiuretic hormone (proAVP) or fragments and partial peptides, in particular copeptin or metaproten II, are determined in the patient to be investigated. However, copeptin or fragments or partial sequences thereof, or posterior lobe hormone transporter II or fragments or partial sequences thereof, are particularly preferred.

The invention further relates to a method for risk stratification of acute coronary syndromes, or an in vitro diagnostic method, for the early diagnosis or differential diagnosis or prognosis of myocardial infarction or angina pectoris, according to one of the above embodiments, wherein a cut-off value (threshold) in the range of 6-20pmol/L of the marker prodiuretic hormone (proAVP) or fragments and partial peptides, in particular copeptin or posterior leaflet hormone transporter II, is important (specific) for the diagnosis and/or risk stratification after the onset of symptoms. Also preferred is a cut-off value (threshold) range of 6-20pmol/L, especially 7.5pmol/L, preferably up to 2 hours after symptom onset.

The invention further relates to a method for risk stratification of acute coronary syndromes, or an in vitro diagnostic method, for the early diagnosis or differential diagnosis or prognosis of myocardial infarction or angina pectoris, according to one of the above embodiments, wherein a cut-off value (threshold) in the range of 10-30pmol/L of the marker prodiuretic hormone (proAVP) or fragments and partial peptides, in particular copeptin or posterior leaflet hormone transporter II, is important (specific) for the prediction and/or risk stratification after the onset of symptoms. Also preferred is a cut-off value (threshold) of 10-20 pmol/L. On this basis, the methods according to the invention advantageously have sensitivity.

In one embodiment of the method according to the invention, a body fluid, in particular blood, optionally whole blood or serum or collectable plasma, is drawn from the patient to be investigated and the diagnosis is carried out ex vivo, that is to say outside the human or animal body. As a result of the determination of the marker prodiuretic hormone (proAVP) or fragments and partial peptides thereof, in particular copeptin or metaproterin II, a high sensitivity and specificity for acute coronary syndrome, myocardial infarction and angina pectoris is achieved and a diagnosis or risk stratification can be carried out on the basis of the amounts present in at least one patient sample. However, markers and peptoids (proAVP or stable fragments of pro-antidiuretic prohormone) or fragments or partial sequences thereof are very particularly preferred. Also particularly preferred is the marker posterior leaflet hormone transporter (proAVP or stable fragment of pro-antidiuretic hormone) or a fragment or partial sequence thereof.

Within the scope of the present invention, "proantidiuretic hormone" is understood as meaning human proteins or polypeptides which are obtainable from proantidiuretic hormone and, for proantidiuretic hormone, comprise amino acids 29 to 164 (see also WO 2006/018315 and FIG. 3) and fragments or partial peptides obtainable therefrom, in particular copeptin (fragment: amino acids 126-164(39 amino acids: SEQ: ASDRSNATQL DGPAGALLLR LVQLAGAPEP FEPAQPDAY) or metafolin transporter II (fragment of proantidiuretic hormone: amino acids 32-124(93 amino acids: SEQ: AMSDLELRQC LPCGPGGKGR CFGPSICCAD ELGCFVGTAE ALRCQEENYLPSPCQSGQKA CGSGGRCAAF GVCCNDESCV TEPECREGFH RRA). The polypeptides according to the invention may also have posttranslational modifications, such as, for example, glycation, lipidation or derivatization.

In a further preferred embodiment, the invention relates to the diagnosis and/or risk stratification and/or early diagnosis or differential diagnosis and/or prognosis of acute coronary syndromes, in particular Acute Myocardial Infarction (AMI) and Angina Pectoris (AP), wherein the metaproterin is determined in the patient to be investigated.

In a further embodiment, proantidiuretic hormone (proAVP) or fragments and partial peptides thereof, in particular copeptin or metaproten II, can also be determined using further markers, preferably those which have been indicated for acute coronary syndromes, in particular myocardial infarction or angina pectoris.

The present invention therefore relates to such an embodiment of the method according to the invention, wherein the determination is also carried out in the patient to be investigated using at least one further marker selected from an inflammation marker, a cardiovascular marker, a neurohormonal marker, or an ischemia marker.

According to the invention, the inflammatory marker may be selected from the group comprising: c-reactive protein (CRP), cytokines such as TNF-alpha, interleukins such as IL-6, procalcitonin (1-116, 3-116), and adhesion molecules such as VCAM or ICAM; and cardiovascular markers, in particular markers indicative of myocardial tissue necrosis and markers affecting blood pressure, may be selected from the group comprising: creatine kinase, myoglobin, myeloperoxidase, natriuretic proteins, in particular ANP (or ANF), proANP, NT-proANP, BNP, proBNP, NT-proBNP or in each case part sequences thereof, cardiac troponin or CRP. Also included are circulation-regulating (pro) hormones, in particular progastrin-releasing peptide (proGRP), pro-endothelin-1 (proendothien-1), pro-leptin (prolein), pro-neuropeptide-Y (proneuropeptide-Y), pro-somatostatin (proaminostatin), pro-neuropeptide-YY, pro-melanocortin, pro-adrenomedullin (proADM), or in all cases partial sequences thereof. The ischemic marker may be selected from at least one marker of the group comprising troponin I and T and CK-MB. Furthermore, the neurohormonal marker can be at least one natriuretic protein, in particular ANP (or ANF), proANP, NT-proANP, BNP, proBNP, NT-proBNP or in each case a partial sequence thereof.

In a particularly preferred embodiment, the invention relates to a particularly advantageous combination of a biomarker and proantidiuretic hormone (proAVP) or fragments or partial peptides thereof, in particular with peptin or the posterior lobe hormone transporter II, with natriuretic protein ANP (or ANF), proANP, NT-proANP, BNP, proBNP, NT-proBNP or in each case partial sequences thereof.

The invention therefore relates to a method for the in vitro diagnosis of acute coronary syndromes, myocardial infarctions or angina pectoris, in which the marker prodiuretic (proAVP) or fragments or partial peptides thereof, in particular copeptin or metaphylogenin II, is determined in combination with a natriuretic protein, in particular ANP (or ANF), proANP, NT-proANP, BNP, proBNP, or NT-proBNP, or in each case a partial sequence thereof, in a patient to be investigated. Again, particular preference is given to the posterior leaflet hormone transporter II, and the combination of peptin with BNP, proBNP, NT-proBNP, in particular with peptin and proBNP.

It is particularly advantageous that the mentioned biomarker combinations have a synergistic effect, which leads to an improved specificity and sensitivity for the diagnosis (see examples).

In a further embodiment of the invention, for in vitro diagnostics the method according to the invention may be performed by means of parallel or simultaneous determination of the markers (e.g. using a multi-titer plate comprising 96 or more wells, wherein the determination is performed for at least one patient sample.

In addition, the method according to the invention and the assays therefore can be carried out in an automatic analyzer-based diagnostic device, in particular using the Kryptor system (r: (r))http://www.kryptor.net/)。

In further embodiments, the methods according to the invention and assays therefor may be carried out as single or multi-parameter assays using a rapid assay (e.g., lateral flow assay). A particularly preferred embodiment comprises a device that is self-detecting or adapted for emergency care diagnostics.

The invention further relates to the use of proantidiuretic hormone (proAVP) or fragments or partial peptides thereof, in particular copeptin or metaproten II, for risk stratification for acute coronary syndromes, myocardial infarction or angina pectoris and/or for the in vitro diagnosis of the early or differential diagnosis or prognosis of myocardial infarction or angina pectoris.

In a particular embodiment, the invention relates to the use of proantidiuretic hormone (proAVP) or fragments or partial peptides thereof, in particular with peptin or vasopressin II, in combination with a natriuretic protein, in particular ANP (or ANF), proANP, NT-proANP, BNP, proBNP, NT-proBNP or in each case a partial sequence thereof, for the diagnosis and/or risk stratification of acute coronary syndromes, myocardial infarctions or angina pectoris.

It is a further object to provide a corresponding diagnostic device for performing the method according to the invention.

Within the scope of the present invention, such diagnostic devices, in particular arrays or assays (e.g. immunoassays, ELISA, etc.), are understood in a broad sense as devices for performing the method according to the invention.

The invention further relates to a kit for risk stratification of acute coronary syndromes, myocardial infarction and/or angina pectoris, comprising detection reagents for the determination of prodiuretic prohormone (proAVP) or fragments or partial peptides thereof, in particular copeptin or metaproterin II, and optionally the additional markers mentioned above. For example, such detection reagents include antibodies.

In a particular embodiment, the invention relates to a kit for the diagnosis and/or risk stratification of acute coronary syndromes, myocardial infarctions and/or angina pectoris, comprising detection reagents for determining the presence of prodiuretic (proAVP) or fragments or partial peptides thereof, in particular copeptin or metaproteoligenin II, in combination with a natriuretic protein, in particular ANP (or ANF), proANP, NT-proANP, BNP, proBNP, NT-proBNP or in each case partial sequences thereof, and optionally further markers as described above. For example, such detection reagents include antibodies.

The following examples and figures are used to explain the invention in more detail, but without limiting the invention to the examples and figures.

Examples and figures: patients entering hospital emergency care with primary symptoms of chest pain have taken blood samples during the initial exam.

131 patients with Myocardial Infarction (MI) (mean age: 64.0 years) were observed for the appearance of acute MI symptoms over a total period of 180 days:

blood samples were taken 3 times within the first 6 hours after the onset of symptoms (chest pain) and hospital admission, each 0-2 hours, 24 hours and 46 hours after the onset of MI symptoms.

In all cases, the copeptin was assayed and NT-proBNP.

After admission, within the first 4 days (days 2-5) after the therapeutic intervention (standard, see Braunwald et al, 2002), in each case one blood sample and subsequent copeptin assay was performed.

After discharge (number of hospitalization days: 7.2 days), the patients were observed during a period of 180 days. The development of severe cardiac insufficiency (cardiac insufficiency) and/or the death of the patient were determined as events. Of the patients observed, 115 had no event (o.E.) and 16 had event (+ E.). The median age of the groups was 63.8 and 64.5 years.

To evaluate the diagnostic effect of biomarkers for early diagnosis of MI and for risk stratification, blood sampling results from a single patient were compared with results from 200 individuals (median age: 65.2 years) who showed no symptoms of MI (control). The results are shown in table 1 and fig. 1 and 2.

Table 1 shows the sensitivity of MI patients (value > cut-off (threshold)) at a given period after the appearance of symptoms, compared to the corresponding specificity of the control (value < cut-off (threshold)).

Within the initial observation period (0-2 hours), copeptin increased greatly compared to the control. For example, only 4.5% of all controls showed copeptin values (specificity 95.5%) greater than 7.5pmol/L (example cut-off (threshold)), while 78.3% of patients without subsequent events, and even 87.5% of patients with events showed copeptin concentrations greater than 7.5pmol/L (sensitivity). This leads to a surprising increase in sensitivity compared to the markers used so far in routine diagnostics (troponin, myoglobin, CK-MB), which then allows these biomarkers to be used for early and differential diagnosis of MI, as well as for risk stratification of acute coronary syndromes (troponin; CK-MB generally occurs only after a 6 hour post-event, whereas myoglobin occurs earlier and has a lower sensitivity, but then decreases after about 2 hours; i.e.a diagnostic window of uncertainty results). Advantageously, the increase in copeptin, i.e., the time of blood draw, is secondary to early diagnosis (e.g., 78.3/80.0/73% sensitivity at 7.5pmol/L cut-off at 0-2/2-4/4-6 hours, respectively). In combination with proBNP or NT-proBNP

Surprisingly, it was shown that the combination of copeptin with proBNP (NT-proBNP) leads to a significant increase in the sensitivity of the early diagnosis of MI (see Table 1). Although proBNP (NT-proBNP) is less effective than copeptin in the assessment of > 7.5pmol/L and/or proBNP (NT-proBNP) > 200pmol/mL, for example at a specificity of approximately 95% comparable to the respective individual marker, it increases the sensitivity by 3-6% at all times in the non-event group and by 6-9.5% in the event group compared to copeptin as individual marker. Table 1:

02h o.E.

02h +E.

24h o.E.

24h +E.

46h o.E.

46h +E.

control (specificity)

Heptamin (pmol/L)

Sensitivity/specificity to cut-off value 6

88.7％

93.8％

89.6％

93.8％

80.0％

87.5％

93.0％

Sensitivity/specificity to cut-off 7.5

78.3％

87.5％

80.0％

87.5％

73.0％

75.0％

95.5％

Sensitivity/specificity to cut-off value 10

67.0％

68.8％

68.7％

68.8％

56.5％

68.8％

97.5％

Sensitivity/specificity for a cut-off of 20 NT-ProBNP (pmol/mL), cut-off 200 combined: copeptin > 7.5pmol/L and/or NT-proBNP > 200pmol/mL

44.3％ 58.5％ 81.2％

68.8％ 62.0％ 93.5％

43.5％ 61.5％ 83.5％

68.8％ 65.0％ 96.0％

40.0％ 66.5％ 79.5％

62.5％ 70.0％ 84.5％

100.0％ 94.5％ 93.8％

Sample recovery, biomarker analysis: blood was collected using standard serum monovites. After 20-40min clotting time, centrifugation was performed at 2000g for 15 min, followed by serum separation by decantation. Serum samples were stored at-20 ℃ for further use.

NT-proBNP was determined using fluorescence immunoassay according to Omland et al (Omland T., Persson A., Ng L., et al., N-terminal pro-B-type natural peptide and long-terminal in acid coronary syndromes, Circulation, 2002; 106: 29132918).

Copeptin was determined using BRAHMS AG's copeptin fluorescence immunoassay. Copeptin assay methods are described in detail in morganthaler n.g. et al, clin.chem.2006 Jan, 52(1), 112119. In summary: 50-microliter of the sample was aspirated into the test tube coated with copeptin antibody (AB1) and mixed with 200 microliter of acridinium ester-labeled anti-copeptin antibody (AB2) solution and incubated at room temperature for 2 hours. After removing unbound (free) labeled antibody by washing 4 times with a washing solution (Lumitest washing solution, Brahms AG), bound acridinium ester labeled antibody is determined in a luminometer of Berthold.

Prognosis (predicted event occurrence): surprisingly, MI patients with events were shown to exhibit even higher copeptin concentrations within 180 days after admission than patients without events. Within 6 hours after the onset of MI symptoms, copeptin showed a relative risk of approximately 1: 2 at an exemplary cut-off (threshold) of 30pmol/L (copeptin > 30 patients had twice the risk of subsequent events compared to < 30pmol/L patients).

As expected, the concentration dropped after the intervention (day 2ff., table 2 b). In this case, it was surprisingly shown that patients with subsequent events showed much less reduction in biomarker. This resulted in a dramatic increase in risk, up to a value of 1: 7.14 for patients with copeptin values above 30 pmol/L. TABLE 2a

02h

02h

24h

24h

46h

46h

+E.

o.E.

+E.

o.E.

+E.

o.E.

Hepeptin

Patients > 30pmol/L are at relative risk of cardiac insufficiency or death up to 180 days after discharge

62.5％ 1.7

36.5％

62.5％ 1.75

35.7％

62.5％ 2.25

27.8％

TABLE 2b

Day 2

Day 2

Day 3

Day 3

Day 4

Day 4

Day 5

Day 5

+E.

o.E.

+E.

o.E.

+E.

o.E.

+E.

o.E.

Hepeptin

Patients > 30pmol/L are at relative risk of cardiac insufficiency or death up to 180 days after discharge

50.0％ 7.14

7.0％

43.8％ 6.26

7.0％

39.2％ 6.42

6.1％

31.3％ 5.13

6.1％

Description in tables and drawings: surv. -survival, e. -event: death, the formation of severe cardiac insufficiency, cutoff in pmol/L, Sens sensitivity, Spef specificity, o.

Posterior leaflet hormone transporter:

according to the earlier description (pullan (supra)): a radioimmunoassay of the posterior lobe hormone transporter was prepared by isolating and quantifying the neurophysin wa isolated and quantified. Rabbits were immunized in this manner, resulting in recovery of high titer anti-metaphylogen transporter antisera. For the immunoassay, the highest titer antisera was used at a concentration of 1: 100,000. Purified posterior leaflet hormone transporter proteins were radioiodinated using the chloramine-T method and used as tracers in assays. Dilutions of purified metafolacin transporter in standard horse serum were used as standards. The analysis was performed as follows: 50- μ L of the sample or standard was mixed with 100 μ L of tracer (12,000dpm, each assay) and 100 μ L of diluted anti-metaplalin antiserum and incubated at 4 ℃ for 24 hours. 100mM sodium phosphate, pH 7.5 and 0.1% BSA were used as buffer. The antibody-bound tracer was separated from free tracer by addition of 60% ethanol followed by centrifugation at 4 ℃ and 5,000g for 15 minutes. The supernatant was discarded and the remaining radioactivity in the pellet was determined. The analysis was performed using Multicalc software. The assay has an assay detection limit of 22 pg/mL. The assay has a measurement range of up to 400 pg/mL. Plasma samples were measured for each patient using this assay, as described below. Samples with measurements > 400pg/mL were measured at appropriate dilutions to give measurements within the measurement range.

Myocardial infarction/diagnosis in 66 patients with myocardial infarction, no more than 6 hours after the onset of myocardial infarction, samples were taken and the posterior folate transporter was measured. For comparison, the posterior folate transporter was determined in200 healthy controls. The receiver performing the characteristic analysis for myocardial infarction diagnosis yielded an AUC of 0.95. For a cutoff of 213pg/mL, a sensitivity of 62.6% was obtained with a specificity of 98%. For a cutoff of 136.1pg/mL, a sensitivity of 84% was obtained at 95% specificity.

Myocardial infarction/prediction (prognosis) in 66 patients with acute myocardial infarction, no more than 6 hours after the onset of myocardial infarction or the second day after infarction, samples were taken and the posterior folate transporter was measured. The patient was observed for a period of 360 days. During this period, 58 patients had no adverse events and 8 patients died or were readmitted due to cardiac insufficiency.

Day one prognosis (< 6 hours after infarct formation occurred): using the receiver for the characteristic analysis, the optimal cut-off value (defined as yielding the maximum sensitivity and specificity) was determined for the prognosis of mortality or readmission due to cardiac insufficiency: 777 pg/mL. At this cut-off, the prognostic sensitivity was 62.5% and the specificity was 73%. At a cutoff value of 777pg/mL, the likelihood ratio of an adverse event was 2.3.

Day 2 prognosis: using the receiver for the characteristic analysis, the optimal cut-off value (defined as yielding the maximum sensitivity and specificity) was determined for the prognosis of mortality or readmission due to cardiac insufficiency: 261 pg/mL. At this cut-off, the prognostic sensitivity was 68.8% and the specificity was 73%. At a cutoff value of 261pg/mL, the likelihood ratio of an adverse event is 2.6.

The attached drawings are as follows: FIG. 1: patient copeptin values after myocardial infarction. Plasma samples were collected from 131 patients at various times post-myocardial infarction, as described. The copeptin values of healthy subjects (control; 200 values) were also plotted. Values for copeptin for each group are plotted as box-and-whisker plots.

FIG. 2: patient copeptin values after myocardial infarction. Plasma samples were collected at various times post myocardial infarction, as described. The patients were then grouped, which indicated the subsequent occurrence of death or readmission due to cardiac insufficiency ("event"; 16 patients) or not ("surv"; 115 patients). Also illustrated are the copeptin values for healthy subjects (control; 200 values). Values for copeptin for each group are plotted as box-and-whisker plots.

FIG. 3: the amino acid sequence of preproproantidiuretic hormone (164 amino acids) is elucidated, as well as partial peptides or fragments of proAVP (amino acids: 29-164), the posterior folate transporter II (amino acids: 32-124) and copeptin (amino acids: 126-164).

Claims (28)

1. Use of a detection reagent for the detection of copeptin or neurophysin II for the preparation of a kit or a diagnostic device for risk stratification of acute coronary syndromes.

2. Use according to claim 1, the kit or diagnostic device being for identifying patients with a high risk and/or poor prognosis of acute coronary syndromes.

3. The use according to claim 2, the kit or diagnostic device for identifying patients with a high risk and/or poor prognosis of myocardial infarction and angina pectoris.

4. The use according to claim 2, wherein the patient is a symptomatic and/or asymptomatic patient.

5. The use of claim 4, wherein the patient is a critical care patient.

6. The use according to claim 1, the kit or diagnostic device for the therapeutic management of acute coronary syndrome.

7. The use according to claim 6, the acute coronary syndrome being myocardial infarction AMI and angina pectoris AP.

8. Use according to claim 6 or 7, the kit or diagnostic device being used in intensive or emergency medical care.

9. The use according to claim 1, the kit or diagnostic device for making clinical decisions.

10. The use according to claim 9, the kit or diagnostic device for further processing with drugs and clinical decision of treatment.

11. The use according to claim 9 or 10, the kit or diagnostic device being used for clinical decision making in intensive or emergency medical care.

12. Use of a detection marker and a detection reagent for pepstatin or metaproterin II for the preparation of a kit or a diagnostic device for the prognosis of myocardial infarction or angina pectoris.

13. Use of a detection reagent for the detection of markers and pepstatin or metaproten II and natriuretic protein or partial sequences thereof for the preparation of a kit or a diagnostic device for the in vitro diagnosis of acute coronary syndrome, myocardial infarction or angina pectoris.

14. Use according to claim 13, said diuretic protein being ANP or ANF, proANP, NT-proANP, BNP, proBNP, NT-proBNP.

15. Use according to any one of claims 13 to 14, wherein the kit or diagnostic device is for prophylaxis, prognosis, differential diagnostic early detection and identification, severity assessment, and progress assessment of concomitant therapy.

16. Use according to claim 1, 12 or 13, characterized in that the kit or diagnostic device further comprises a detection reagent for at least one additional marker selected from an inflammation marker, a cardiovascular marker, a neurohormonal marker or an ischemia marker.

17. Use according to claim 16, wherein the marker of inflammation is at least one marker selected from the group consisting of: c-reactive protein CRP, cytokines, interleukins, procalcitonin, and adhesion molecules.

18. Use according to claim 17, wherein the marker of inflammation is at least one marker selected from the group consisting of: the cytokines TNF-alpha, interleukin IL-6, procalcitonin 1-116, procalcitonin 3-116 and adhesion molecules VCAM or ICAM.

19. Use according to claim 16, wherein the cardiovascular marker is at least one marker selected from the group consisting of: creatine kinase, myoglobin, myeloperoxidase, natriuretic protein or partial sequences thereof, cardiac troponin, CRP, and circulation-regulating hormones or procirculation-regulating hormones, or in each case partial sequences thereof.

20. Use according to claim 19, characterized in that the cardiovascular marker is at least one marker selected from the group consisting of: ANP or ANF, proANP, NT-proANP, BNP, proBNP, NT-proBNP, the progastrin-releasing peptide proGRP, pro-endothelin-1, pro-leptin, pro-neuropeptide-Y, pro-somatostatin, pro-neuropeptide-YY, pro-melanocortin and pro-adrenomedullin proADM, or in each case a partial sequence thereof.

21. Use according to claim 16, characterized in that the ischemia marker is at least one marker selected from the group comprising troponin I and T, CK-MB.

22. The use according to claim 16, wherein the neurohormonal marker is at least one natriuretic protein, or a partial sequence thereof.

23. Use according to claim 22, wherein said natriuretic protein is ANP or ANF, proANP, NT-proANP, BNP, proBNP, NT-proBNP.

24. Use according to claim 1, 12 or 13, characterized in that the kit or diagnostic device is used for the determination of at least one patient sample.

25. Use according to claim 24, wherein the assay is performed on an automated analyzer.

26. The use of claim 25, wherein the automated analyzer is Kryptor.

27. Use according to claim 24, characterized in that the determination is carried out using an accelerated method.

28. The use of claim 27, wherein the assay is a single-parameter or multi-parameter assay.