Abstract
The present invention is directed to methods for determining active DPP3 in a bodily fluid sample, an assay or kit for determining active DPP3 in a bodily fluid sample, a method for diagnosing a disease or condition in a subject accompanied by or related to necrotic processes and methods of treating or preventing said disease.
Claims
1-41 (canceled)
42. A method for treating a disease or condition in a subject accompanied by or related to necrotic processes, wherein said disease is selected from the group consisting of heart failure, chronic heart failure, acute heart failure (AHF), myocardial infarction (MI), stroke, liver failure, burn injuries, traumatic injuries, severe infection, severe infection that is microbial, severe infection that is viral, AIDS, severe infection that is parasitic, malaria, SIRS, sepsis, cancer, acute kidney injury (AKI), CNS disorders, seizures, neurodegenerative diseases, autoimmune diseases, vascular diseases, Kawasaki syndrome or hypotension, comprising administering to a subject in need thereof an effective amount of an inhibitor of the activity of DPP3.
43. The method according to claim 42, wherein said inhibitor is an anti-DPP3 antibody or anti-DPP3 antibody fragment or anti-DPP3 non-Ig scaffold.
44. The method according to claim 42, wherein said inhibitor is an antibody that is mono-binding or at least two-binding.
45. The method according to claim 42, wherein said inhibitor is an anti-DPP3 antibody or anti-DPP3 antibody fragment or anti-DPP3 non-Ig scaffold that binds to SEQ ID No. 1.
46. The method according to claim 42, wherein said inhibitor is an anti-DPP3 antibody or anti-DPP3 antibody fragment or anti-DPP3 non-Ig scaffold that binds to SEQ ID No. 2.
47. The method according to claim 42, wherein said inhibitor is an antibody or fragment or scaffold that exhibits a minimum binding affinity to DPP3 of equal or less than 10.sup.7M.
48. The method according to claim 42, wherein said inhibitor is an antibody or fragment or scaffold that is monospecific.
49. The method according to claim 42, wherein said inhibitor is an antibody or fragment or scaffold binds to full-length DPP3 and inhibits activity of DPP3 of at least 10%, or at least 50%, more preferred at least 60%, even more preferred more than 70%, even more preferred more than 80%, even more preferred more than 90%, even more preferred more than 95%.
50. The method according to claim 42, wherein said inhibitor is selective and/or specific to DPP3 and not cross cell membranes and/or the blood brain barrier.
51. The method according to claim 42, wherein said subject has an amount of total DPP3 and/or an amount of active DPP3 in a sample of bodily fluid of said subject that is above a predetermined threshold.
52. The method according to claim 42, wherein the inhibitor of the activity of DPP3 is in a pharmaceutical composition.
53. The method according to claim 42, wherein a method is involved of extracorporeal removal of DPP3 from plasma comprising apheresis and affinity chromatography.
54. The method according to claim 42, wherein the disease is selected from the group consisting of acute heart failure (AHF), myocardial infarction (MI), liver failure, burn injuries, severe infection, severe infection that is microbial, severe infection that is viral, AIDS, severe infection that is parasitic, malaria, SIRS, sepsis, cancer, and acute kidney injury (AKI).
55. The method according to claim 42, wherein the disease is hypotension.
Description
BRIEF DESCRIPTION OF THE DRAWINGS
[0415] FIG. 1A: Illustrates the Inhibition of DPP3 activity: The activity of recombinant GST-hDPP3 was measured in the presence of several different antiDPP3 antibodies. DPP3 binding antibodies, that were produced against peptides and/or full-length (FL) native DPP3, show a strong inhibitory effect of up to 70%.
[0416] FIG. 1B: Illustrates the Inhibition of DPP3 activity: Inhibition curve of recombinant GST-hDPP3 with inhibitory mAbDPP3. Inhibition of DPP3 by a specific antibody is concentration dependent, with an IC.sub.50 at 0.2 g/ml.
[0417] FIG. 2A: Illustrates the DPP3 concentration as diagnostic marker: DPP3 concentration in EDTA plasma of healthy controls and patients with various diseases (AHFacute heart failure, MImyocardial infarct, sepsis, cancer, AKIacute kidney injury, LRTIlower respirational tract infection). Medians of patient groups differ significantly from healthy controls (Mann-Whitney test p<0.005).
[0418] FIG. 2B: Illustrates the DPP3 concentration as diagnostic marker: Comparison of plasma DPP3 concentrations of patients that died shortly within admission to the emergency department and surviving patients. Surviving patients show significantly lower DPP3 levels (Mann-Whitney test p<0.05).
[0419] FIG. 3: Illustrates the DPP3 activity as diagnostic marker: DPP3 activity in EDTA plasma of healthy controls and patients with various diseases (AHFacute heart failure, sepsis, AMacute kidney injury, LRTIlower respirational tract infection). Medians of patient groups differ significantly from healthy controls (Mann-Whitney test p<0.0001).
[0420] FIG. 4A: Illustrates the ROC plot analysis of the DPP3 activity and concentration assay: ROC analysis of healthy controls and patients suffering from AHF.
[0421] FIG. 4B: Illustrates the ROC plot analysis of the DPP3 activity and concentration assay: ROC analysis of healthy controls and patients suffering from sepsis.
[0422] FIG. 5: Illustrates the Safety of mAbDPP3 treatment (blood pressure): Healthy rats treated with PBS or mAbDPP3 (5.75 mg/kg). Blood pressure (BP) was measured and recorded via a catheter inserted into the Arteria carotis communis dextra. The administration and sampling catheter were inserted into the Vena jugularis sinistra. Treatment of slightly increases relative blood pressure compared to PBS treated rats (n=3 per group).
[0423] FIG. 6: Illustrates the Influence of mAbDPP3 on mortality of septic mice: Septic mice (CLP model) were treated with PBS or mAbDPP3 (1,9 mg/kg) 5 minutes before and 2 h after CLP. Mortality was monitored over 7 days. The Kaplan-Meyer plot shows increased survival of septic mice after mAbDPP3 treatment.
[0424] FIG. 7A: Illustrates the Influence of mAbDPP3 on heart failure of septic rats: Experimental design of heart failure study of rats in septic shock.
[0425] FIG. 7B: Illustrates the Influence of mAbDPP3 on heart failure of septic rats: CLP induces heart failure in rats, as indicated by a decreased shortening fraction compared to sham animals. This shortening fraction is significantly increased by mAbDPP3 treatment (2 mg/kg; n7 per group; Mann-Whitney test p<0.0001).
[0426] FIG. 7C: Illustrates the Influence of mAbDPP3 on heart failure of septic rats: Mean blood pressure of vehicle treated septic rats decreases with time whereas mAbDPP3 treatment leads to a significant increase in mBP (2 mg/kg; n7 per group; Mann-Whitney test p<0.005).
[0427] FIG. 8: Illustrates the Influence of mAbDPP3 on tumor growth in vitro: Soft-Agar Assay with tumor cell lines (lung, colon and breast cancer). Addition of antiDPP3 antibody reduces tumor cell growth.
[0428] FIG. 9A: Illustrates the Influence of mAbDPP3 on tumor growth in vivo: Mice with xenograft of breast tumor cells (n=10 per group) were treated with PBS or mAbDPP3. Growth curve of relative tumor volume over 24 days shows a decreased tumor growth in mAbDPP3 treated mice.
[0429] FIG. 9B: Illustrates the Influence of mAbDPP3 on tumor growth in vivo: Comparison of time the breast cell tumor needs to 20-fold increase its volume with and without mAbDPP3 treatment. Growth takes significantly longer with mAbDPP3 treatment (Mann-Whitney test, p<0.05).
[0430] FIG. 9C: Illustrates the Influence of mAbDPP3 on tumor growth in vivo: Mice with xenograft of colon tumor cells (n=10 per group) were treated with PBS or mAbDPP3. Growth curve of relative tumor volume over 30 days shows a decreased tumor growth in mAbDPP3 treated mice.
[0431] FIG. 9D: Illustrates the Influence of mAbDPP3 on tumor growth in vivo: Comparison of time the colon cell tumor needs to 10-fold increase its volume with and without mAbDPP3 treatment. Growth takes longer with mAbDPP3 treatment.
[0432] FIG. 10: Illustrates the DPP3 activity as diagnostic marker (II): DPP3 activity in EDTA plasma of healthy controls and patients with various diseases (acute myocardial infarction (AMI), cardiogenic shock, septic shock and liver failure). Medians of patient groups differ significantly from healthy controls (Mann-Whitney test p<0.05).
[0433] FIG. 11: Illustrates the Effect of DPP3 on blood pressure in healthy rats: Healthy male Wistar rats were injected with 0.2 mg/kg recombinant GST-hDPP3. Blood pressure (BP) was measured and recorded via a catheter inserted into the Arteria carotis communis dextra. DPP3 was injected i.v. via the tail vain. DPP3 injection leads to decreased BP.
[0434] FIG. 12A: Illustrates Peptide and small molecule inhibitors of DPP3.
[0435] FIG. 12B: Illustrates Further Peptide and small molecule inhibitors of DPP3.
[0436] FIG. 12 C: Illustrates Even Further Peptide and small molecule inhibitors of DPP3.
[0437] FIG. 13: Illustrates Immunogen sequence, designation and characteristics of produced anti-DPP3 antibodies.
[0438] FIG. 14: Illustrates Comparison of DPP3 values of diseased patients and healthy controls in a sandwich type immune assay.
[0439] FIG. 15: Illustrates Comparison of DPP3 values of diseased patients and healthy controls in a sandwich type immune assay and in an enzyme capture activity assay.
[0440] FIG. 16: Illustrates Data of ROC analysis (AUCarea under the curve; CIconfidence interval).
[0441] FIG. 17: Illustrates Experimental groups.
[0442] FIG. 18: Illustrates Overview of treatment strategy.
[0443] FIG. 19: Illustrates Concentrations of native or recombinant DPP3 in samples before and after affinity chromatography with depicted anti-DPP3 antibodies.
