Treatment of patients infected with coronavirus with proadrenomedullin or its fragments, and conjugates for adrenomedullin.

Abstract

The subject of the present invention is a method for diagnosing or predicting the risk of a life-threatening deterioration or adverse event, or for prognosing the severity, or for predicting or monitoring the success of a therapeutic or interventional treatment in a patient infected with coronavirus, the method comprising: measuring the level of proadrenomedullin (SEQ ID NO: 31) or a fragment thereof in a body fluid sample from said patient; comparing the level of proadrenomedullin or a fragment thereof with a predetermined threshold or a previously determined level of proadrenomedullin or a fragment thereof, and correlating the level of said proadrenomedullin or fragment thereof with the risk of a life-threatening deterioration or adverse event; or correlating the level of said proadrenomedullin or fragment thereof with severity, or correlating the level of said proadrenomedullin or fragment thereof with the success of the treatment or intervention; or The proadrenomedullin or a fragment thereof is selected from the group consisting of PAMP (SEQ ID NO: 32), MR-proADM (SEQ ID NO: 33), ADM-NH2 (SEQ ID NO: 20), ADM-Gly (SEQ ID NO: 21), and CT-proADM (SEQ ID NO: 34). The subject of the present invention is an anti-adrenomedullin (ADM) antibody or an anti-ADM antibody fragment or an anti-ADM non-Ig scaffold for use in the therapeutic or interventional treatment of patients infected with coronavirus.

Description

[Technical Field]

[0001] The subject matter of the present invention is a method for (a) diagnosing or predicting the risk of life-threatening exacerbation or adverse events in patients infected with coronavirus, or (b) diagnosing or determining the severity or prognosis, or (c) predicting or monitoring the success of treatment or intervention, or (d) providing guidance on treatment or stratifying treatment, or (e) managing patients, and these methods are - A step of measuring the level of proadrenomedullin (SEQ ID NO: 31) or its fragments in a body fluid sample from the patient. - A step of comparing the level of proadrenomedullin or its fragment with a predetermined threshold or a previously measured level of proadrenomedullin or its fragment, and - A process of correlating the level of proadrenomedullin or its fragments with the risk of life-threatening exacerbation or adverse events, or - A step of correlating the level of proadrenomedullin or its fragments with the severity, or The process of correlating the level of proadrenomedullin or its fragments with the success of a therapeutic or interventional treatment, The process of correlating the level of proadrenomedullin or its fragments with a specific treatment or intervention, or The process includes correlating the level of proadrenomedullin or its fragments with the management of the patient, The proadrenomedullin or its fragment is selected from the group consisting of PAMP (SEQ ID NO: 32), MR-proADM (SEQ ID NO: 33), ADM-NH2 (SEQ ID NO: 20), ADM-Gly (SEQ ID NO: 21), and CT-proADM (SEQ ID NO: 34).

[0002] The subject of this invention is an anti-adrenomedullin (ADM) antibody, an anti-ADM antibody fragment, or an anti-ADM non-Ig scaffold used for the treatment or interventional therapy of patients infected with coronavirus. [Background Technology]

[0003] The peptide adrenomedullin (ADM) was first described in 1993 as a novel antihypertensive peptide containing 52 amino acids (Kitamura et al., 1993. Biochem Biophys Res Comm 192 (2): 553-560), and was isolated from a human pheochromocytoma cell line (SEQ ID NO: 20). In the same year, the cDNA encoding a precursor peptide containing 185 amino acids, and the entire amino acid sequence of this precursor peptide, were also described. In particular, the precursor peptide containing a 21-amino acid signal sequence at the N-terminus is called "pre-pro-adrenomedullin" (pre-proADM). In this specification, all identified amino acid positions are usually related to pre-proADM containing 185 amino acids. Subsequently, pre-proADM is converted to pro-ADM (SEQ ID NO: 31) containing 164 amino acids by cleavage of the N-terminal signal peptide. pro-ADM is further processed to obtain the 20 peptides at the N-terminus of pro-ADM (PAMP; SEQ ID NO: 32), the central region pro-ADM (MR-proADM; SEQ ID NO: 33), adrenotensin pro-ADM 153-185 (CT-pro ADM; SEQ ID NO: 34), and immature ADM, as well as C-terminal glycine-extended ADM (ADM-Gly; SEQ ID NO: 21). This is then converted to the mature, bioactive form of ADM (bio-ADM; ADM-NH2; SEQ ID NO: 20) by enzymatic amidation of its C-terminus. More than half of known neuropeptides and endocrine peptides require the formation of a C-terminal α-amide group to achieve full bioactivity (Guembe et al. 1999. J Histochem Cytochem 47(5): 623-36; Vishwanatha et al. 2013. Handbook of Biologically Active Peptides Peptidylglycine Amidating Monoxygenase (PAM). Second Edi. Elsevier Inc.). This final step in peptide hormone biosynthesis involves the action of peptidylglycine α-amide-forming monooxygenase (PAM), a bifunctional enzyme that specifically recognizes the C-terminal glycine (CT-Gly) residue in its substrate.PAM cleaves glyoxylic acid from the peptide CT-Gly residue in a two-step enzymatic reaction, leading to the production of an α-amidated peptide hormone at the C-terminus, with the resulting α-amide group derived from the cleaved CT-Gly (Prigge et al. 2000. Cellular and Molecular Life Sciences 57(8): 1236-59). This amidation reaction occurs in the lumen of the secretory granules prior to the exocytosis of the amidation product (Martinez et al. 1996. Am J Pathol 149(2): 707-16).

[0004] The discovery and characterization of ADM in 1993 spurred intensive research, the results of which are summarized in various review articles cited within the context of this specification, with particular reference to the "peptide" literature specifically on ADM (Takahashi 2001. Peptides 22: 1691; Eto 2001. Peptides 22: 1693-1711). Further reviews include those by Hinson et al. in 2000 (Hinson et al. 2000. Endocrine Reviews 21(2): 138-167). Scientific studies to date have shown that ADM can be considered, in particular, a multifunctional regulatory peptide. As mentioned above, it is released into the circulating system in an inactive form amplified by glycine (Kitamura et al. 1998. Biochem Biophys Res Commun 244(2): 551-555). There are also binding proteins that are specific to ADM and are thought to similarly modulate the effects of ADM (Pio et al. 2001. The Journal of Biological Chemistry 276(15): 12292-12300). To date, the most important physiological effect of ADM and PAMP has been their effect on blood pressure.

[0005] Therefore, ADM is an effective vasodilator, and its antihypertensive effect can be attributed to a specific peptide compartment in the C-terminal region of ADM. Furthermore, the aforementioned bioactive peptide PAMP, generated from pre-pro-ADM, appears to have a different mechanism of action than ADM, but has been found to exhibit a similar antihypertensive effect (see also Eto et al. 2001 and Hinson et al. 2000, Kuwasaki et al. 1997. FEBS Lett 414(1): 105-110; Kuwasaki et al. 1999. Ann. Clin. Biochem. 36: 622-628; Tsuruda et al. 2001 Life Sci. 69(2): 239-245 and EP-A2 0 622 458 in addition to the review above). Furthermore, it has been found that ADM concentrations measurable in circulating fluids and other bodily fluids are significantly higher in some pathological conditions than in healthy control subjects. Therefore, ADM levels are significantly elevated, to varying degrees, in patients with congestive heart failure, myocardial infarction, kidney disease, hypertensive diseases, diabetes, the acute phase of shock, and sepsis and septic shock. PAMP concentrations are also elevated in some of the aforementioned pathological conditions, although plasma levels are lower compared to ADM (Eto 2001. Peptides 22: 1693-1711). High concentrations of ADM have been observed in sepsis, and the highest concentrations have been reported in septic shock (Eto 2001. Peptides 22: 1693-1711; Hirata et al. Journal of Clinical Endocrinology and Metabolism 81(4): 1449-1453; Ehlenz et al. 1997. Exp Clin Endocrinol Diabetes 105: 156-162; Tomoda et al. 2001. Peptides 22: 1783-1794; Ueda et al. 1999. Am. J. Respir. Crit. Care Med. 160: 132-136; Wang et al. 2001. Peptides 22: 1835-1840).Furthermore, plasma concentrations of ADM are elevated in heart failure patients and correlate with disease severity (Hirayama et al. 1999. J Endocrinol 160: 297-303; Yu et al. 2001. Heart 86: 155-160). High plasma ADM is an independent negative prognostic indicator in these patients (Poyner et al. 2002. Pharmacol Rev 54: 233-246).

[0006] Kitamura et al. reported that the concentrations of mature ADM and ADM-Gly were significantly elevated in the plasma of hypertensive patients compared to healthy volunteers (Kitamura et al. 1998. Biochem Biophys Res Comm 244(2): 551-5). In both groups, mature ADM was much lower than ADM-Gly. However, there was no significant difference in the ratio of mature ADM to ADM-Gly between hypertensive and non-hypertensive patients.

[0007] In the early stages of sepsis, ADM has been reported to improve cardiac function as well as blood supply to the liver, spleen, kidneys, and small intestine. Anti-ADM neutralizing antibodies neutralize these effects during the early stages of sepsis (Wang et al. 2001. Peptides 22: 1835-1840). In other diseases, inhibition of ADM may be beneficial to some extent. However, complete neutralization of ADM may be harmful because some physiological functions may require a certain amount of ADM. Many reports emphasize that ADM administration may be beneficial for certain diseases. In contrast, other reports state that ADM is life-threatening when administered under certain conditions.

[0008] WO 2013 / 072510 discloses a non-neutralizing N-terminal anti-ADM antibody for use in the treatment of severe chronic or acute illnesses or acute symptoms in patients to reduce the patient's risk of death.

[0009] WO 2013 / 072511 discloses a non-neutralizing N-terminal anti-ADM antibody for use in the treatment of chronic or acute diseases or acute symptoms in patients for the prevention or reduction of organ dysfunction or organ failure.

[0010] WO 2013 / 072513 discloses an N-terminal anti-ADM antibody for use in the treatment of acute illnesses or symptoms in patients to stabilize the circulatory system.

[0011] WO 2013 / 072514 discloses an N-terminal anti-ADM antibody for regulating fluid balance in patients with chronic or acute diseases or acute symptoms.

[0012] WO 2019 / 154900 discloses a non-neutralizing N-terminal anti-ADM antibody for use in the treatment and prevention of dementia. Furthermore, WO 2019 / 154900 discloses a method for diagnosing dementia and monitoring its (preventive) treatment by measuring the ratio of the level of mature ADM to the level of proadrenomedullin or its fragments.

[0013] WO 2013 / 072512 is a report on the half-life (t) of adrenomedullin in serum, blood, and plasma. 1 / 2 This document discloses a non-neutralizing N-terminal anti-ADM antibody, which is an ADM-stabilizing antibody that enhances the half-retention time.

[0014] The efficacy of non-neutralizing antibodies targeting the N-terminus of ADM was investigated in survival studies of CLP-induced sepsis in mice. Preliminary treatment with non-neutralizing antibodies reduced catecholamine infusion, improved renal dysfunction, and improved overall survival (Struck et al. 2013. Intensive Care Med Exp 1(1):22; Wagner et al. 2013. Intensive Care Med Exp 1(1):21). Furthermore, antibodies against the central region of ADM (MR-ADM antibodies) also significantly improved survival in mice with CLP-induced sepsis, although to a lesser extent than N-terminal anti-ADM antibodies (Struck et al. 2013. Intensive Care Med Exp 1(1):22).

[0015] These positive results led to the development of a humanized N-terminal anti-ADM antibody, named adrecizumab, for further clinical application. The beneficial effects of adrecizumab on vascular barrier function and survival have recently been demonstrated in preclinical models of systemic inflammation and sepsis (Geven et al. 2018. Shock 50(6): 648-654). In this study, pretreatment with adrecizumab reduced renal vascular leakage in rats with endotoxemia and mice with CLP-induced sepsis, consistent with increased renal expression of the protective peptide Ang-1 and decreased expression of the harmful peptide vascular endothelial growth factor. Furthermore, pretreatment with adrecizumab improved the 7-day survival rate in mice with CLP-induced sepsis by 10–50% with a single dose and 0–40% with repeated doses. Furthermore, Phase I studies demonstrated excellent safety and tolerability as follows: no serious adverse events were observed, no signals of adverse events more frequently occurring in subjects treated with adrecizumab were detected, and no changes related to other safety parameters were found (Geven et al. 2017. Intensive Care Med Exp 5 (Suppl 2): ​​0427). Of particular interest is the disclosed mechanism of action of adrecizumab. Data from both animals and humans reveal a potent dose-dependent increase in circulating ADM after administration of this antibody. Based on pharmacokinetic data and the lack of increase in MR-proADM (an inactive peptide fragment derived from the same prohormone as ADM), the higher circulating ADM levels cannot be explained by increased production.

[0016] Although ADM is small enough to cross the endothelial barrier, its antibodies are not, so the mechanistic explanation for this increase may be that excess antibodies in the circulatory system may excrete ADM from the interstitium into the circulatory system (Geven et al. 2018. Shock. 50(2): 132-140). Furthermore, antibody binding to ADM leads to an extension of ADM's half-life. NT-ADM antibodies partially inhibit ADM-mediated signaling, and when circulatory ADM increases significantly, they increase ADM activity in blood compartments "net" overall, where they exert beneficial effects on EC (primarily barrier stabilization), while reducing the harmful effects of ADM on interstitial VSMC (vasodilation).

[0017] In other words, when functional plasma ADM levels are elevated, NT-ADM antibodies are hypothesized to target vascular and capillary leakage based on sepsis and inflammation. The latter can lead to exacerbation from severe COVID-19 to septic shock and ARDS (Veerdonk et al. 2020. Preprints, 2020040023 (doi: 10.20944 / preprints202004.0023.v1)). More recently, vascular endothelial stabilization has been clearly identified as a therapeutic target for COVID-19 (Varga et al. 2020.395(10234):1417-1418).

[0018] An N-terminal ADM antibody named adrecizumab (HAM8101) was administered to eight critically ill COVID-19 patients with acute respiratory spurt syndrome (ARDS) (Karakas et al. 2020. Biomolecules 10: 1171). Patients received a single dose of adrecizumab, administered 1–3 days after the initiation of mechanical ventilation. Median SOFA (12.5) and SAPS-II (39) scores clearly indicated that this group was at the highest risk. Follow-up periods ranged from 13 to 27 days. Following adrecizumab administration, one patient in the low-dose group died on day 4 from fulminant pulmonary embolism, but four remained stable and three were discharged from the intensive care unit (ICU). Within 12 days, SOFA scores and disease severity scores (ranging from 0 to 16, reflecting critical resources in the ICU, with higher scores indicating greater severity) decreased in 5 out of 7 surviving patients (among all high-dose patients). PaO2 / FiO2 increased within 12 days, but inflammatory parameters such as C-reactive protein, procalcitonin, and interleukin-6 decreased. Importantly, mortality was lower than predicted and calculated by the SOFA score. In conclusion, in this preliminary, uncontrolled series of 8 shock patients with life-threatening COVID-19 and ARDS, adrecizumab administration demonstrated favorable outcomes.

[0019] Coronaviruses are widespread in humans and several other vertebrates, causing respiratory, intestinal, hepatic, and neurological illnesses. In particular, the severe acute respiratory syndrome coronavirus (SARS-CoV) of 2003 and the Middle East respiratory syndrome coronavirus (MERS-CoV) of 2012 caused outbreaks in humans. Comparisons with SARS-CoV reveal several notable differences and similarities. Both MERS-CoV and SARS-CoV have significantly higher mortality rates (40% and 10%, respectively) (de Wit et al. 2016. SARS and MERS: recent insights into emerging coronaviruses. Nat Rev Microbiol 14(8):523-34; Zhou et al. 2020. A pneumonia outbreak associated with a new coronavirus of probable bat origin. Nature 579(7798):270-273). The current SARS-CoV-2 shares 79% of its genome with SARS-CoV, but is thought to be far more transmissible. Both SARS-CoV strains enter cells via the angiotensin-converting enzyme 2 (ACE2) receptor (Wan et al. 2020. Receptor recognition by novel coronavirus from Wuhan: An analysis based on decade-long structural studies of SARS. Nature 579(7798):270-2730). The disease caused by SARS-CoV-2 is called coronavirus disease 2019 (COVID-19).

[0020] SARS-CoV-2 first mainly infects the lower respiratory tract and binds to ACE2 on the surface of alveolar epithelial cells. Both viruses are potent inducers of inflammatory cytokines. "Cytokine storm" or "cytokine cascade" is a putative mechanism of organ damage. This virus activates immune cells and induces the secretion of inflammatory cytokines and chemokines into the pulmonary vascular endothelium.

[0021] The clinical spectrum of SARS-CoV-2 infection appears to be broad, ranging from asymptomatic infection, mild upper respiratory tract disease, severe viral pneumonia with respiratory failure, and even death, with many patients being hospitalized due to pneumonia (Huang et al. 2020. Clinical features of patients infected with 2019 novel coronavirus in Wuhan, China. "Clinical features of patients infected with 2019 novel coronavirus in Wuhan, China." Lancet 395: 497-506; Wang et al. 2020. Clinical characteristics of 138 hospitalized patients with 2019 novel coronavirus-infected pneumonia in Wuhan, China. "Clinical characteristics of 138 hospitalized patients with 2�19 novel coronavirus-infected pneumonia in Wuhan, China." JAMA 323(11):1061-1069; Chen et al. 2020. Epidemiological and clinical characteristics of 99 cases of 2019 novel coronavirus pneumonia in Wuhan, China: a descriptive study. "Epidemiological and clinical characteristics of 99 cases of 2019 novel coronavirus pneumonia in Wuhan, China: a descriptive study." Lancet 395: 507-1)

[0022] Very recently, clinicians have proposed advanced age, elevated D-dimer levels, and high SOFA scores to identify early patients with COVID-19 with poor prognosis (Zhou et al. 2020. Clinical course and risk factors for mortality of adult inpatients with COVID-19 in Wuhan, China: a retrospective cohort study. "Clinical course and risk factors for mortality of adult inpatients with COVID-19 in Wuhan, China: a retrospective cohort study" The Lancet, 395(10229):1054-1062).

[0023] The subject matter of the present invention is a method for diagnosing or predicting (a) the risk of life-threatening deterioration or adverse events, or (b) diagnosing or prognosticating severity, or (c) predicting or monitoring the success of treatment or interventional treatment, or (d) guiding treatment or stratifying treatment, or (e) managing patients in a patient infected with a coronavirus, the method comprising · measuring the level of proadrenomedullin (SEQ ID NO: 31) or a fragment thereof in a body fluid sample of the patient, · comparing the level of proadrenomedullin or a fragment thereof with a predetermined threshold value of proadrenomedullin or a fragment thereof or a previously measured level, and · correlating the level of proadrenomedullin or a fragment thereof with the risk of life-threatening deterioration or adverse events, or · correlating the level of proadrenomedullin or a fragment thereof with severity, or · correlating the level of proadrenomedullin or a fragment thereof with the success of treatment or interventional treatment, or · correlating the level of proadrenomedullin or a fragment thereof with a specific treatment or interventional treatment, or The process includes correlating the levels of proadrenomedullin or its fragments with the management of the patient, The proadrenomedullin or its fragment is selected from the group consisting of PAMP (SEQ ID NO: 32), MR-proADM (SEQ ID NO: 33), ADM-NH2 (SEQ ID NO: 20), ADM-Gly (SEQ ID NO: 21), and CT-proADM (SEQ ID NO: 34).

[0024] The subject of the present invention is a method for (a) diagnosing or predicting the risk of life-threatening exacerbation or adverse events in patients infected with coronavirus, or (b) diagnosing or determining the severity or prognosis, or (c) predicting or monitoring the success of treatment or interventional therapy, or (d) providing guidance or stratifying treatment, or (e) managing patients, wherein the coronavirus is selected from the group including SARS-CoV-1, SARS-CoV-2, and MERS-CoV, and is particularly SARS-CoV-2.

[0025] The subject matter of the present invention is a method for (a) diagnosing or predicting the risk of life-threatening exacerbation or adverse events in patients infected with coronavirus, or (b) diagnosing or predicting the severity or prognosis, or (c) predicting or monitoring the success of treatment or interventional therapy, or (d) providing guidance on treatment or stratifying treatment, or (e) managing patients, wherein the adverse events are selected from a group including death, organ failure, shock, ARDS, and ALI (acute lung injury).

[0026] The subject matter of the present invention is a method for (a) diagnosing or predicting the risk of life-threatening exacerbation or adverse events in patients infected with coronavirus, or (b) diagnosing or predicting the severity or prognosis, or (c) predicting or monitoring the success of treatment or interventional therapy, or (d) providing guidance or stratifying treatment, or (e) managing patients, wherein the level of proadrenomedullin or a fragment thereof is above a predetermined threshold.

[0027] In a particular embodiment of the present invention, the level of proadrenomedullin or its fragments is measured at least twice.

[0028] In another specific embodiment of the present invention, the at least second measurement of the level of proadrenomedullin or its fragments is taken within 2 hours, preferably within 4 hours, more preferably within 6 hours, even more preferably within 12 hours, even more preferably within 24 hours, and most preferably within 48 hours.

[0029] This means that, as is obvious throughout the subject matter of the invention, according to the term “previously measured level of proadrenomedullin or its fragment,” this previously measured level is the level measured within 2 hours, preferably within 4 hours, more preferably within 6 hours, even more preferably within 12 hours, even more preferably within 24 hours, and most preferably within 48 hours. The difference between the measured value and the previous measured value is the relative difference in the said level of proadrenomedullin or its fragment in different samples taken from the patient at different time points.

[0030] An increase of 70 pg / mL or more, or a bio-ADM increase of 25% or more (a minimum increase of 50 pg / mL), will persist until the end of the following day.

[0031] In another specific embodiment of the present invention, the levels of proadrenomedullin or its fragments are measured in different samples taken from the patient at different time points in time.

[0032] In another specific embodiment of the present invention, the difference between the levels of proadrenomedullin or its fragments in different samples taken from the patient at different time points is measured. The difference may be measured as an absolute difference or a relative difference.

[0033] In another specific embodiment of the present invention, treatment is initiated when the relative difference between the levels of proadrenomedullin or its fragments in different samples taken from the patient at different time points is 100% or more, more preferably 75% or more, even more preferably 50% or more, and most preferably 25% or more.

[0034] In another specific embodiment of the present invention, treatment is initiated when, in the second, i.e., additional measurement, the relative level of proadrenomedullin or its fragment is at least 25%, and the absolute level of proadrenomedullin or its fragment is at least 50 pg / mL, wherein the proadrenomedullin fragment is mature ADM (ADM-NH2).

[0035] The subject of the present invention is a method for (a) diagnosing or predicting the risk of life-threatening exacerbation or adverse events in a patient infected with coronavirus according to the present invention, or (b) diagnosing or predicting the severity or prognosis, or (c) predicting or monitoring the success of treatment or interventional treatment, or (d) guiding or stratifying treatment, or (e) managing a patient, wherein the fragment is MR-proADM (SEQ ID NO: 33), and a predetermined threshold for MR-proADM in the subject body fluid sample is 0.5 to 2 nmol / L, preferably 0.7 to 1.5 nmol / L, preferably 0.8 to 1.2 nmol / L, and most preferably a threshold of 1 nmol / L.

[0036] The subject of the present invention is a method for (a) diagnosing or predicting the risk of life-threatening exacerbation or adverse events in a patient infected with coronavirus according to the present invention, or (b) diagnosing or predicting the severity or prognosis, or (c) predicting or monitoring the success of treatment or interventional treatment, or (d) providing guidance or stratifying treatment, or (e) managing a patient, wherein the fragment is ADM-NH2 (SEQ ID NO: 20), and the predetermined threshold for ADM-NH2 (SEQ ID NO: 20) in the subject body fluid sample is 40 to 100 pg / mL, more preferably 50 to 90 pg / mL, even more preferably 60 to 80 pg / mL, and most preferably the threshold is 70 pg / mL.

[0037] The subject matter of the present invention is a method for (a) diagnosing or predicting the risk of life-threatening exacerbation or adverse events in a patient infected with coronavirus according to the present invention, or (b) diagnosing or predicting the severity or prognosis, or (c) predicting or monitoring the success of treatment or interventional treatment, or (d) providing guidance or stratifying treatment, or (e) managing a patient, wherein the patient's SOFA score is 3 or higher, preferably 7 or higher, or the quick SOFA score is 1 or higher, preferably 2 or higher.

[0038] The subject of the present invention is a method for (a) diagnosing or predicting the risk of life-threatening exacerbation or adverse events in a patient infected with coronavirus according to the present invention, or (b) diagnosing or predicting the severity or prognosis, or (c) predicting or monitoring the success of treatment or interventional therapy, or (d) providing guidance or stratifying treatment, or (e) managing a patient, wherein the patient has a D-dimer level of 0.5 μg / mL or higher, preferably 1 μg / mL or higher.

[0039] The subject matter of the present invention is a method for (a) diagnosing or predicting the risk of life-threatening exacerbation or adverse events in patients infected with coronavirus according to the present invention, or (b) diagnosing or predicting the severity or prognosis, or (c) predicting or monitoring the success of treatment or interventional therapy, or (d) directing or stratifying treatment, or (e) managing patients, wherein the level of proadrenomedullin or its fragments is measured by contacting the body fluid sample with a capture agent that specifically binds to proadrenomedullin or its fragments.

[0040] The subject matter of the present invention is a method for (a) diagnosing or predicting the risk of life-threatening exacerbation or adverse events in patients infected with coronavirus according to the present invention, or (b) diagnosing or determining the severity or prognosis, or (c) predicting or monitoring the success of treatment or interventional therapy, or (d) guiding or stratifying treatment, or (e) managing patients, wherein the measurement comprises the use of a capture agent that specifically binds to proadrenomedullin or a fragment thereof, and the capture agent may be selected from the group consisting of antibodies, antibody fragments, or non-IgG scaffolds.

[0041] The subject matter of the present invention is a method for (a) diagnosing or predicting the risk of life-threatening exacerbation or adverse events in a patient infected with coronavirus according to the present invention, or (b) diagnosing or predicting the severity or prognosis, or (c) predicting or monitoring the success of treatment or interventional therapy, or (d) directing or stratifying treatment, or (e) managing a patient, wherein the level of proadrenomedullin or a fragment thereof is measured in a body fluid sample of the subject, the measurement comprising the use of a capture binder that specifically binds to proadrenomedullin or a fragment thereof, the capture binder being an antibody.

[0042] The subject matter of the present invention is a method for (a) diagnosing or predicting the risk of life-threatening exacerbation or adverse events in a patient infected with coronavirus according to the present invention, or (b) diagnosing or predicting the severity or prognosis, or (c) predicting or monitoring the success of treatment or interventional treatment, or (d) directing or stratifying treatment, or (e) managing a patient, wherein the level of proadrenomedullin or a fragment thereof is measured in a body fluid sample of the subject, the measurement comprising the use of a scavenging agent that specifically binds to the level of proadrenomedullin or a fragment thereof, the scavenging agent being immobilized on the surface.

[0043] The subject of the present invention is a method for (a) diagnosing or predicting the risk of life-threatening exacerbation or adverse events in a patient infected with coronavirus according to the present invention, or (b) diagnosing or predicting the severity or prognosis, or (c) predicting or monitoring the success of treatment or interventional treatment, or (d) directing or stratifying treatment, or (e) managing a patient, wherein the patient is treated with an anti-adrenomedullin (ADM) antibody, an anti-ADM antibody fragment, or an anti-ADM non-Ig scaffold, the anti-ADM antibody, anti-ADM fragment, or anti-ADM non-Ig scaffold conjugated to the N-terminal and / or central region (amino acids 1-42) of ADM-Gly and / or ADM-NH2: YRQSMNNFQGLRSFGCRFGTCTVQKLAHQIYQFTDKDKDNVA (SEQ ID NO: 23).

[0044] The subject of this invention is an anti-adrenomedullin (ADM) antibody, an anti-ADM antibody fragment, or an anti-ADM non-Ig scaffold used for the treatment or interventional therapy of patients infected with coronavirus.

[0045] The subject of the present invention is an anti-adrenomedullin (ADM) antibody, an anti-ADM antibody fragment, or an anti-ADM non-Ig scaffold for use in the treatment or interventional therapy of a patient infected with coronavirus according to the present invention, wherein the coronavirus is selected from the group including Sars-CoV-1, Sars-CoV-2, and MERS-CoV, and is particularly Sars-CoV-2.

[0046] The subject of the present invention is an anti-adrenomedullin (ADM) antibody, an anti-ADM antibody fragment, or an anti-ADM non-Ig scaffold for use in the treatment or intervention therapy of a patient infected with coronavirus according to the present invention, wherein the patient has a level of proadrenomedullin or its fragment in a body fluid sample of the subject that, when measured by a method according to the above-described technique, exceeds a predetermined threshold or is higher than a previously measured level of proadrenomedullin or its fragment.

[0047] The subject of the present invention is an anti-adrenomedullin (ADM) antibody, an anti-ADM antibody fragment, or an anti-ADM non-Ig scaffold for use in the treatment or interventional therapy of a patient infected with coronavirus according to the present invention, wherein the patient's SOFA score is 3 or higher, preferably 7 or higher, or the rapid SOFA score is 1 or higher, preferably 2 or higher.

[0048] The subject of the present invention is an anti-adrenomedullin (ADM) antibody, an anti-ADM antibody fragment, or an anti-ADM non-Ig scaffold for use in the treatment or intervention therapy of a patient infected with coronavirus according to the present invention, the patient having a D-dimer level of 0.5 μg / mL or higher, preferably 1 μg / mL or higher.

[0049] The subject of the present invention is an anti-adrenomedullin (ADM) antibody, an anti-ADM antibody fragment, or an anti-ADM non-Ig scaffold for use in the treatment or intervention therapy of patients infected with coronavirus according to the present invention, wherein the anti-ADM antibody, anti-ADM fragment, or anti-ADM non-Ig scaffold is bound to the N-terminal portion (amino acids 1-21) of ADM-Gly and / or ADM-NH2: YRQSMNNFQGLRSFGCRFGTC (SEQ ID NO: 14).

[0050] The subject of the present invention is an anti-adrenomedullin (ADM) antibody, an anti-ADM antibody fragment, or an anti-ADM non-Ig scaffold for use in the treatment or intervention therapy of patients infected with coronavirus according to the present invention, wherein the anti-adrenomedullin (ADM) antibody, an anti-ADM antibody fragment, or an anti-ADM non-Ig scaffold is 10 times the value of pro-adrenomedullin or its fragment. -7 It exhibits a minimum binding affinity of M or less.

[0051] The subject of the present invention is an anti-adrenomedullin (ADM) antibody, an anti-ADM antibody fragment, or an anti-ADM non-Ig scaffold used for the treatment or intervention therapy of patients infected with coronavirus according to the present invention, wherein the anti-adrenomedullin (ADM) antibody, anti-ADM antibody fragment, or anti-ADM non-Ig scaffold inhibits the bioactivity of ADM by 80% or less, preferably 50% or less.

[0052] The subject of the present invention is an anti-adrenomedullin (ADM) antibody, an anti-ADM antibody fragment, or an anti-ADM non-Ig scaffold for use in the treatment or intervention therapy of patients infected with coronavirus according to the present invention, wherein the antibody is a monoclonal antibody or a monoclonal antibody fragment.

[0053] The subject of this invention is an anti-adrenomedullin (ADM) antibody, an anti-ADM antibody fragment, or an anti-ADM non-Ig scaffold for use in the treatment or intervention therapy of patients infected with coronavirus according to this invention, wherein the complementarity-determining region (CDR) within the heavy chain comprises the following sequence: CDR1: Sequence ID 1 GYTFSRYW CDR2: Sequence ID 2 ILPGSGST CDR3: Sequence ID 3 TEGYEYDGFDY Furthermore, the complementarity-determining region (CDR) within the light chain contains the following sequence: CDR1: Sequence ID 4 QSIVYSNGNTY CDR2: RVS CDR3: Sequence ID 5 FQGSHIPYT.

[0054] The subject of the present invention is an anti-adrenomedullin (ADM) antibody, an anti-ADM antibody fragment, or an anti-ADM non-Ig scaffold for use in the treatment or intervention therapy of patients infected with coronavirus according to the present invention, wherein the antibody or fragment is a sequence selected from the group comprising the following sequences as the VH region: Sequence number 6 (AM-VH-C): [ka] Sequence ID 7 (AM-VH1): [ka] Sequence ID 8 (AM-VH2-E40): [ka] Sequence ID 9 (AM-VH3-T26-E55): [ka] Sequence ID 10 (AM-VH4-T26-E40-E55): [ka] Or it includes sequences that are more than 80% identical to each of the above sequences, and Sequences selected from the group containing the following sequences as the VL region: Sequence ID 11(AM-VL-C): [ka] Sequence ID 12 (AM-VL1): [ka] Sequence ID 13 (AM-VL2-E40): [ka] Or include sequences that are more than 80% identical to each of the sequences mentioned above.

[0055] The subject of the present invention is an adrenomedullin (ADM) antibody, an anti-ADM antibody fragment, or an anti-ADM non-Ig scaffold for use in the treatment or intervention therapy of patients infected with coronavirus according to the present invention, wherein the antibody or fragment has the following sequence as its heavy chain: Sequence ID 35: [ka] or containing sequences that are more than 95% identical to it, Furthermore, the light chain has the following sequence: Sequence ID 36: [ka] Or it includes sequences that are more than 95% identical to it.

[0056] The subject of the present invention is an anti-adrenomedullin (ADM) antibody, an anti-ADM antibody fragment, or an anti-ADM non-Ig scaffold for use in the treatment or intervention therapy of a patient infected with coronavirus according to the present invention, wherein the monoclonal antibody or antibody fragment is a humanized monoclonal antibody or a humanized monoclonal antibody fragment.

[0057] The subject of the present invention is an anti-adrenomedullin (ADM) antibody, an anti-ADM antibody fragment, or an anti-ADM non-Ig scaffold for use in the treatment or intervention therapy of patients infected with coronavirus according to the present invention, wherein the anti-adrenomedullin (ADM) antibody, anti-ADM antibody fragment, or anti-ADM non-Ig scaffold is a monoclonal antibody and is adrecizumab, and has the following sequence as its heavy chain: Sequence ID 35: [ka] And the following sequence as the light chain: Sequence ID 36: [ka] It contains or is a biosimilar thereof.

[0058] In accordance with the present invention, the body fluid is a blood sample in a particular embodiment. The blood sample may be selected from the group including whole blood, serum, and plasma. In a particular embodiment of the diagnostic method, the sample is selected from the group including human citrate plasma, heparin plasma, and EDTA plasma.

[0059] The concentration of a biomarker such as a D-dimer, such as proadrenomedullin or a fragment thereof, may be measured by an immunoassay, which may be a sandwich immunoassay, preferably a fully automated assay.

[0060] In one embodiment, the test sensitivity of the assay method for ADM-Gly is such that it can quantify ADM-Gly in healthy subjects, and its sensitivity is 20 pg / mL, preferably 15 pg / mL, and more preferably 10 pg / mL.

[0061] In one embodiment, the test sensitivity of the above-mentioned assay method for PAMP is such that it can quantify PAMP in healthy subjects, and its sensitivity is less than 0.5 pmol / L, preferably less than 0.25 pmol / L, and more preferably less than 0.1 pmol / L.

[0062] In one embodiment, the test sensitivity of the assay method for detecting CT-proADM is such that it can quantify CT-proADM in healthy subjects, and its sensitivity is less than 100 pmol / L, preferably less than 75 pmol / L, and more preferably less than 50 pmol / L.

[0063] In one embodiment, the test sensitivity of the assay method for detecting ADM-NH2 is such that it can quantify ADM-NH2 in healthy subjects, and its sensitivity is less than 70 pg / mL, preferably less than 40 pg / mL, and more preferably less than 10 pg / mL.

[0064] In one embodiment, the test sensitivity of the above-mentioned assay method is capable of quantifying MR-proADM in healthy subjects, and its sensitivity is less than 0.5 nmol / L, preferably less than 0.4 nmol / L, and more preferably less than 0.2 nmol / L.

[0065] In addition to proadrenomedullin and / or its fragments, further biomarkers may be measured. These further biomarkers may be selected from the group including D-dimer, procalcitonin (PCT), C-reactive protein (CRP), lactate, DPP3, penKid, NT-proBNP, white blood cell count, lymphocyte count, neutrophil count, hemoglobin, platelet count, albumin, alanine aminotransferase, creatinine, serum urea, lactate dehydrogenase, creatinine kinase, cardiac troponin I, prothrombin time, serum ferritin, interleukin-6 (IL-6), IL-10, IL-2, IL-7, tumor necrosis factor-α (TNF-α), granulocyte colony-stimulating factor (GCSF), IP-10, MCP-1, and MIP-1α.

[0066] Another embodiment of this application relates to an anti-ADM antibody or anti-ADM antibody fragment or anti-ADM non-Ig scaffold for use in the treatment of a patient, wherein the anti-ADM antibody or anti-ADM fragment or anti-ADM non-Ig scaffold is bound to the N-terminal and / or central region (amino acids 1-42) of ADM-Gly and / or ADM-NH2: YRQSMNNFQGLRSFGCRFGTCTVQKLAHQIYQFTDKDKDNVA (SEQ ID NO: 23).

[0067] One embodiment of this application relates to an anti-ADM antibody, an anti-ADM antibody fragment, or an anti-ADM non-Ig scaffold for use in the treatment of patients infected with coronavirus, wherein the anti-ADM antibody, anti-ADM antibody fragment, or anti-ADM non-Ig protein scaffold is intended for the following purposes: a. To be used in the treatment of the patient in order to stabilize the patient's systemic circulation. Here, the patient needs to have his / her systemic circulation stabilized and has a heart rate greater than 100 beats / min and / or a mean arterial pressure less than 65 mmHg, and stabilizing the systemic circulation means raising the mean arterial pressure to greater than 65 mmHg; or b. To be used to prevent an increase in heart rate to over 100 beats / min and / or a decrease in mean arterial pressure to less than 65 mmHg in patients infected with coronavirus.

[0068] Another embodiment of the present application relates to an anti-ADM antibody or anti-ADM antibody fragment or anti-ADM non-Ig scaffold for use in the treatment of a patient infected with coronavirus, wherein the anti-ADM antibody or anti-ADM antibody fragment or anti-ADM non-Ig protein scaffold is intended for use in the treatment of the patient to prevent or mitigate organ failure or to prevent organ failure, the organ being selected from the group including the heart, kidney, liver, lung, pancreas, small intestine, and spleen.

[0069] In a particular embodiment of the present invention, the patient has been diagnosed with or is suspected of having a coronavirus infection.

[0070] The term "coronavirus infection" is defined as an infection caused by coronaviruses (coronavirus family), a family of single-stranded RNA viruses with a positive envelope. The viral genome is 26–32 kilobases long. The particles are typically decorated with large (about 20 nm) club-shaped or petal-shaped surface protrusions ("peplomers" or "spikes"), and electron micrographs of spherical particles form images reminiscent of the sun's corona. Coronaviruses cause disease in mammals and birds. In humans, the virus causes respiratory infections such as the common cold, which are typically mild, but rare forms such as SARS, MERS, and COVID-19 can be fatal. SARS-CoV-2 has been added as the most recent form.

[0071] In certain embodiments, the coronavirus infection is selected from a group that includes infections by SARS-CoV-1, SARS-CoV-2, and MERS-CoV, and is particularly limited to infections by SARS-CoV-2.

[0072] According to the WHO, a severe acute respiratory infection (SARI) suspected to be caused by SARS-CoV-2 infection is currently defined as an acute respiratory infection (ARI) that has a history of fever (i.e., a measured body temperature of 38°C or higher) and cough, has developed within the last 10 days, and requires hospitalization. However, the absence of fever does not rule out viral infection.

[0073] SARS-CoV infection can present with mild, moderate, or severe illness, including severe pneumonia, ARDS, sepsis, and septic shock. Early identification of patients with severe symptoms (see Table 1) allows for optimized, immediate adjuvant therapy and safe, rapid admission (or referral) to the intensive care unit, in accordance with facility or national procedures. In patients with mild symptoms, hospitalization may not be necessary unless there is a concern of rapid deterioration. All patients discharged home should be instructed to return to the hospital if any worsening of their illness occurs. [Table 1-1] [Table 1-2]

[0074] Septic shock is a potentially fatal condition that occurs when sepsis, which is organ failure or damage in response to infection, leads to dangerously low blood pressure and cellular metabolic abnormalities. The Third International Consensus Definitions for Sepsis (Sepsis-3) defines septic shock as a subtype of sepsis in which particularly severe circulatory, cellular, and metabolic abnormalities are associated with a higher mortality risk than sepsis alone. Patients with septic shock can be clinically identified by the need for vasopressors to maintain mean arterial pressure above 65 mmHg and serum lactate levels above 2 mmol / L (greater than 18 mg / dL) without hypovolemia. This combination is associated with a hospital mortality rate of over 40% (Singer et al. 2016. JAMA. 315 (8): 801-10). Primary infections are most commonly caused by bacteria, but can also be caused by fungi, viruses, or parasites. The infection can be located in any part of the body, but most commonly in the lungs, brain, urinary tract, skin, or abdominal organs. This infection can lead to multiple organ failure syndrome (formerly known as multiple organ failure) and can be fatal. In many cases, patients with septic shock are treated in an intensive care unit. Because their immune systems are not as efficient at dealing with infections as healthy adults, children, immunocompromised individuals, and the elderly most commonly suffer from this infection. The mortality rate from septic shock is approximately 25–50%.

[0075] The severity of a disease is defined as the degree of organ dysfunction or physiological decompensation in the patient. Severity may be classified into various stages using a scoring system or the like.

[0076] As used herein, organ dysfunction means a medical condition or health state in which an organ fails to perform its expected function. “Organ dysfunction” means organ dysfunction to the extent that normal homeostasis cannot be maintained without external clinical intervention. Such organ dysfunction may relate to organs selected from the group including the kidneys, liver, heart, lungs, and nervous system. In contrast, organ function refers to the expected function of each organ within its physiological range. Those skilled in the art will know the function of each organ in a health examination.

[0077] Organ dysfunction may be defined by a set of organ dysfunction assessment scores (SOFA scores) or their components. The SOFA score, formerly known as the sepsis-associated organ dysfunction assessment score (Singer et al. 2016. JAMA 315(8): 801-10), is used to track the condition of subjects staying in the intensive care unit (ICU) and measure the degree or rate of organ dysfunction in the subject. The score is based on six different scores, increasing from 0 to 4 for each of the respiratory, cardiovascular, hepatic, coagulation, renal, and nervous systems, reflecting the worsening of organ dysfunction. The evaluation criteria for the SOFA score are described, for example, by Lambden et al. (see Lambden et al. 2019. Critical Care 23: 374 for a review). The SOFA score may traditionally be calculated upon admission to the ICU and every 24 hours thereafter. In particular, the organ dysfunction is selected from a group that includes renal impairment, cardiac dysfunction, hepatic dysfunction, or airway dysfunction.

[0078] The rapid SOFA score (quickSOFA or qSOFA) was introduced in February 2016 by the Sepsis-3 group as a simplified version of the SOFA score, and the first method to identify patients at high risk of poor outcomes due to infection (Angus et al. 2016. Critical Care Medicine. 44 (3): e113-e121). qSOFA dramatically simplifies the SOFA score by including only three clinical criteria and including "any change in mental state" instead of requiring a GCS score of less than 15. qSOFA can be given to patients concisely, quickly, and repeatedly. Scores range from 0 to 3 points. 1 point is awarded for hypotension (SBP ≤ 100 mmHg), high respiratory rate (≥ 22 breaths / min), and change in mental state (GCS ≤ 15). The presence of qSOFA scores of 2 or higher close to the onset of infection was associated with a higher risk of death or prolonged stay in the intensive care unit. These outcomes are more common in infected patients who may have sepsis than in infected patients without complications. Based on these findings, the third internationally agreed definition of sepsis recommends qSOFA as a concise input request to identify infected patients likely to be sepsis outside the ICU (Seymour et al. 2016. JAMA 315(8): 762-774).

[0079] Life-threatening deterioration is defined as a patient condition at high risk of death that involves failure of vital organ systems, including central nervous system disorders, renal failure, hepatic failure, metabolic failure, or respiratory failure.

[0080] Adverse events are defined as death, organ failure or shock, ARDS, and ALI (acute lung injury).

[0081] In this invention, the terms "prognosis" or "making a prognosis" mean predicting how the condition of a subject (e.g., a patient) will progress. This may include the possibility of recovery, the possibility of adverse events, or an estimate of the subject's outcome.

[0082] The prognosis assessment for adverse events, including death, may be carried out over a defined period, for example, up to one year, preferably up to six months, more preferably up to three months, more preferably up to ninety days, more preferably up to sixty days, more preferably up to 28 days, more preferably up to fourteen days, more preferably up to seven days, and more preferably up to three days.

[0083] In certain embodiments, the prognosis for adverse events, including death, is determined over a period of up to 28 days.

[0084] In the context of this invention, the term "treatment monitoring" refers, for example, to monitoring and / or adjusting the treatment of the patient by obtaining feedback regarding the efficacy of the treatment.

[0085] As used herein, the term “treatment guidance” refers to the administration of a specific treatment or medical intervention based on the numerical values ​​of one or more biomarkers and / or clinical parameters and / or clinical scores.

[0086] The aforementioned clinical parameters or clinical scores are selected from a group that includes a history of hypotension, need for vasopressors, intubation, mechanical ventilation, Horowitz index, SOFA score, and rapid SOFA score.

[0087] The term "treatment stratification" refers to grouping or classifying patients into different groups, particularly those receiving or not receiving treatment methods according to their classification.

[0088] The aforementioned treatments or interventions may be selected from a group that includes drug therapy, non-invasive ventilation, mechanical ventilation, extracorporeal membrane oxygenation (ECMO), dialysis, or renal replacement therapy.

[0089] Non-invasive mechanical ventilation is the use of respiratory support delivered through a face mask, nasal mask, or helmet. Typically, oxygenated air is delivered through the mask under positive pressure.

[0090] Mechanical ventilation, or ventilator-assisted ventilation, is a medical term for artificial respiration in which mechanical means are used to assist or replace spontaneous breathing. This may require a machine called a ventilator, or it may involve a qualified professional, such as an anesthesiologist, respiratory therapist (RT), registered nurse, or paramedic, manually assisting breathing by compressing a bag-valve-mask device. Mechanical ventilation is called "invasive" when any device is taken into the trachea through the mouth, such as an endotracheal tube, or through the skin, such as a tracheostomy tube. Face masks or nasal masks are used for non-invasive ventilation in appropriately selected conscious patients.

[0091] Extracorporeal membrane oxygenation (ECMO), also known as extracorporeal life support (ECLS), is an extracorporeal technology that provides long-term cardiac and respiratory support to patients whose hearts and lungs cannot provide enough gas exchange or perfusion to sustain life. ECMO technology is primarily derived from cardiopulmonary bypass and provides short-term support by suspending the original circulatory system. ECMO works by drawing blood from the patient's body, artificially removing carbon dioxide, and adding oxygen to the patient's red blood cells. Generally, it is used either after cardiopulmonary bypass or in the later stages of treatment for patients with severe heart and / or pulmonary failure, but is now also used in certain institutions as a treatment for cardiac arrest, allowing treatment of the underlying cause while circulatory and oxygenation are supported. ECMO is also used to support patients with acute viral pneumonia associated with COVID-19 when ventilators are insufficient to maintain blood oxygenation levels.

[0092] The aforementioned drug therapies may be selected from the group comprising anti-ADM antibodies, anti-ADM antibody fragments, anti-ADM non-Ig scaffolds, antiviral drugs, immunoglobulins from recovered COVID-19 pneumonia patients, neutralizing monoclonal antibodies targeting coronavirus, immunostimulants, camostat mesylate, coronavirus protease inhibitors (such as chymotrypsin-like inhibitors and papain-like protease inhibitors), spike (S) protein angiotensin-converting enzyme 2 (ACE2) blockers (such as chloroquine, hydroxychloroquine, emodin, and promazine), angiotensin receptor agonists, and / or their precursors.

[0093] The neutralizing monoclonal antibodies targeting SARS-CoV and MERS-CoV may be selected from the group summarized by Shanmugaraj et al. (Shanmugaraj et al. 2020. Asian Pac J. allergy Immunol 38: 10-18).

[0094] The aforementioned antiviral drug may be selected from the group including lopinavir, ritonavir, remdesivir, nafamostat, ribavirin, oseltamivir, penciclovir, acyclovir, ganciclovir, favipiravir, nitazoxanide, nelfinavir, and arbidol.

[0095] The immunostimulant may be selected from the group including interferon, intravenous gamma globulin, thymosin α-1, levamisole, and non-immunosuppressive derivatives of cyclosporine-A.

[0096] In one embodiment, the angiotensin receptor agonist and / or its precursor is selected from the group including angiotensin I, angiotensin II, angiotensin III, and angiotensin IV.

[0097] The Horowitz index (synonyms: post-Horowitz oxygenation, Horowitz quotient, P / F ratio) is a ratio used to assess lung function in patients, especially those on mechanical ventilation. It is useful for assessing the degree of lung damage. The Horowitz index is defined as the ratio of the partial pressure of oxygen in the blood (PaO2), expressed in millimeters of mercury, to the fraction of oxygen in the inhaled air (FiO2), i.e., the PaO2 / FiO2 ratio. In healthy lungs, the Horowitz index is age-dependent and is usually between 350 and 450. A value below 300 is the threshold for mild lung damage, and 200 indicates moderate to severe lung damage. A value below 100 is the criterion for severe damage. The Horowitz index plays an important role in the diagnosis of acute respiratory distress syndrome (ARDS). The three severity levels of ARDS are classified according to Berlin's definition (Matthay et al. 2012. J Clin Invest. 122(8): 2731-2740), using the Horowitz index based on the degree of hypoxemia.

[0098] Acute respiratory spurt syndrome (ARDS) is a type of respiratory failure characterized by the rapid onset of widespread inflammation in the lungs. Symptoms include shortness of breath, rapid breathing, and a bluish skin color. A reduced quality of life is common among surviving patients. Causes may include sepsis, pancreatitis, trauma, pneumonia, and aspiration. Underlying mechanisms include diffuse damage to the cells that form the barrier of the lung's micro-air sacs, surfactant dysfunction, activation of the immune system, and dysfunction of the body's blood clotting regulation. In essence, ARDS impairs the lungs' ability to exchange oxygen and carbon dioxide. Diagnosis is based on a PaO2 / FiO2 ratio (ratio of arterial oxygen partial pressure to inspired oxygen fraction) of less than 300 mmHg despite positive end-expiratory pressure (PEEP) above 5 cmH2O. Primary treatment includes mechanical ventilation along with treatment directed at the underlying cause. Methods of ventilation include the use of low-dose and low-pressure ventilators. If oxygenation remains insufficient, lung replacement therapy and neuromuscular blocking agents may be used. If these are insufficient, extracorporeal membrane oxygenation (ECMO) may be an option. This syndrome is associated with a 35-50% mortality rate.

[0099] As used herein, the term “patient” refers to a living human or non-human organism that is receiving or should receive medical treatment for an illness. This also includes humans who are not clearly ill but are being investigated for signs of illness. Accordingly, the methods and measurements described herein are applicable to both human and animal diseases. In the context of this invention, the term "patient management" refers to the following: • Decision to admit to a hospital or intensive care unit, • Decision to transfer the patient to a specialized hospital or specialized hospital department. • Evaluation of early discharge from the intensive care unit or hospital. • Allocation of resources (e.g., physicians and / or nursing staff, diagnostic methods, treatments), and • Decisions regarding treatment.

[0100] Threshold levels can be obtained, for example, from Kaplan-Meier analysis, which correlates disease incidence with the quartile of a biomarker in a population. According to this analysis, subjects with biomarker levels above the 75th percentile have a significantly higher risk of developing the disease according to the present invention. This result is further supported by Cox regression analysis fully adjusted for classical risk factors. The highest quartile for all other subjects is very significantly associated with an increased risk of developing the disease according to the present invention.

[0101] Other preferred cutoff values ​​include, for example, the 90th, 95th, or 99th percentile of the normal population. Using a percentile higher than the 75th percentile can reduce the number of false positives identified, but increases the risk of missing moderately risky cases. Therefore, the cutoff value may be chosen depending on whether it is considered more appropriate to identify the majority of risky cases at the expense of identifying "false positives," or whether it is considered more appropriate to identify primarily high-risk cases at the expense of missing some moderately risky cases.

[0102] The threshold values ​​described above may differ for other measurement methods if those methods are calibrated in a different way than the measurement system used in this invention. Therefore, the aforementioned threshold values ​​should be adjusted accordingly for measurement methods that have been calibrated differently, taking into account the differences in calibration. One possibility for quantifying the differences in calibration is to perform a comparative analysis (correlation) of the test in question (e.g., the bio-ADM test) using both methods for each biomarker (e.g., bio-ADM) in the sample. Another possibility is to measure the test in question by comparing the median biomarker levels of a representative normal population with the median biomarker levels described in the literature (e.g., Weber et al. 2017. JALM 2(2): 222-233), assuming that the test has sufficient analytical sensitivity, and then recalculating the calibration based on the difference obtained from this comparison. Using the calibration employed in this invention, samples from normal (healthy) subjects were measured, and the median plasma bio-ADM (mature ADM-NH2) was 13.7 pg / mL (interquartile range [IQR]: 9.6–18.7 pg / mL) (Weber et al. 2017. JALM 2(2): 222-233).

[0103] Throughout this specification, the “antibody,” “antibody fragment,” or “non-Ig scaffold” according to the present invention can bind to ADM and thus be directed toward ADM, and thus can be referred to as “anti-ADM antibody,” “anti-ADM antibody fragment,” or “anti-ADM non-Ig scaffold.”

[0104] Mature ADM, bio-ADM, and ADM-NH2 are used synonymously throughout this application and are molecules according to Sequence ID No. 20.

[0105] In a particular embodiment of the diagnostic method, the binder is provided with at least 10 proadrenomedullin or its fragments (not ADM-NH2 according to SEQ ID NO: 20) and ADM-NH2. 7 M -1 Preferably 10 8 M -1 Preferably 109 M -1 super, most preferably 10 10 M -1 exhibits super binding affinity. Those skilled in the art will recognize that higher doses of the compound can be used to compensate for lower affinity, and it is understood that this means does not fall outside the scope of the claims of the present invention.

[0106] To measure the affinity of the antibody for adrenomedullin, the kinetics of the binding of adrenomedullin to the immobilized antibody were measured by label-free surface plasmon resonance using a Biacore 2000 system (GE Healthcare Europe GmbH, Freiburg, Germany). Reversible immobilization of the antibody was performed using an anti-mouse Fc antibody covalently bound at high density to the CM5 sensor surface according to the manufacturer's instructions (mouse antibody capture kit; GE Healthcare) (Lorenz et al. 2011. Antimicrob Agents Chemother. 55 (1): 165 - 173).

[0107] In certain embodiments of the diagnostic method, one assay is used to measure the levels of proadrenomedullin or its fragments and ADM - NH2, and the levels of proadrenomedullin or its fragments are selected from the group consisting of PAMP (SEQ ID NO: 32), MR - proADM (SEQ ID NO: 33), ADM - Gly (SEQ ID NO: 21), and CT - proADM (SEQ ID NO: 34), and such an assay is a sandwich assay, preferably a fully automated assay.

[0108] In one embodiment of the present invention, the assay may be a so - called POC assay (point - of - care test), which is an inspection technique that can perform the test within 1 hour near the patient without requiring a fully automated measurement system. An example of this technique is immunochromatographic test technology.

[0109] In one embodiment of the diagnostic method, the assay is a sandwich immunoassay using any type of detection technique, including but not limited to enzyme labeling, chemiluminescence labeling, and electrochemiluminescence labeling, preferably a fully automated assay. In one embodiment of the diagnostic method, the assay is an enzyme-labeled sandwich assay. Examples of automated or fully automated assays include assays that may use one of the systems of Roche Elecsys, Abbott Architect, Siemens Centauer, Brahms Kryptor, BiomerieuxVidas, and Alere Triage (all registered trademarks).

[0110] Various immunoassay methods are known and may be used in the assay methods and procedures of the present invention, including radioimmunoassay ("RIA"), homogeneous enzyme amplification immunoassay ("EMIT"), enzyme-linked immunosorbent assay ("ELISA"), apoenzyme reactivation immunoassay ("ARIS"), dipstick immunoassay, and immunochromatography.

[0111] In a particular embodiment of the diagnostic method, at least one of the two binders is labeled for detection.

[0112] Monospecificity means that the antibody, antibody fragment, or non-Ig scaffold binds to one specific region within the target ADM that comprises at least four amino acids. The monospecific antibody, fragment, or non-Ig scaffold according to the present invention is an antibody, fragment, or non-Ig scaffold that all have affinity for the same antigen. While monoclonal antibodies are monospecific, monospecific antibodies may also be produced by methods other than generating them from common germ cells.

[0113] The anti-ADM antibody or antibody fragment that binds to ADM, or the non-Ig scaffold that binds to ADM, may be a non-neutralizing anti-ADM antibody or antibody fragment that binds to ADM, or a non-neutralizing non-Ig scaffold that binds to ADM.

[0114] The antibody or fragment according to the present invention is a protein comprising one or more polypeptides substantially encoded by an immunoglobulin gene that specifically binds to an antigen. Recognized immunoglobulin genes include the constant region genes of κ, λ, α (IgA), γ (IgG1, IgG2, IgG3, IgG4), δ (IgD), ε (IgE), and μ (IgM), as well as numerous immunoglobulin variable region genes. Full-length immunoglobulin light chains are generally about 25 kd or 214 amino acids in length.

[0115] Full-length immunoglobulin heavy chains are generally about 50 kd or 446 amino acids long. The light chain is encoded by a variable region gene at the NH2 terminus (about 110 amino acids long) and a constant region gene at the COOH terminus (κ or λ). The heavy chain is similarly encoded by a variable region gene (about 116 amino acids long) and one of the other constant region genes.

[0116] The basic structural unit of an antibody is generally a tetramer consisting of two identical pairs of immunoglobulin chains, each pair having one light chain and one heavy chain. In each pair, the variable regions of the light and heavy chains bind to the antigen, while the constant region mediates effector function. Immunoglobulins also exist in various other forms, such as Fv, Fab, and (Fab')2, as well as in bifunctional hybrid antibodies and single chains (e.g., Lanzavecchia et al. 1987. Eur. J. Immunol. 17:105; Huston et al. 1988. Proc. Natl. Acad. Sci. USA, 85: 5879-5883; Bird et al. 1988. Science 242: 423-426; Hood et al. 1984, Immunology, Benjamin, NY, 2nd ed.; Hunkapiller and Hood 1986. Nature 323: 15-16). The variable regions of the light or heavy chains of immunoglobulins include a framework region interrupted by three hypervariable regions, also known as complementarity measurement regions (CDRs) (see Sequences of Proteins of Immunological Interest, E. Kabat et al. 1983, US Department of Health and Human Services). As mentioned above, CDRs are primarily responsible for binding to the antigen's epitope. Immune complexes are antibodies such as monoclonal antibodies, chimeric antibodies, humanized antibodies or human antibodies, or functional antibody fragments that are specifically bound to the antigen.

[0117] Chimeric antibodies are antibodies in which the light and heavy chain genes are typically constructed by genetic engineering from the variable and constant regions of immunoglobulins belonging to various species. For example, the variable compartment of a gene derived from a mouse monoclonal antibody can bind to human constant compartments such as κ and γ1 or γ3. Thus, in one example, a therapeutic chimeric antibody is a variable or antigen-binding domain derived from a mouse antibody and a constant or effector domain derived from a human antibody, but other mammalian species may be used, or the variable region may be generated by molecular technology. Methods for generating chimeric antibodies are well known in the art; see, for example, US Patent No. 5,807,715. "Humanized" immunoglobulins are immunoglobulins containing a human framework region and one or more CDRs derived from non-human immunoglobulins (such as mouse, rat, or synthetic). The non-human immunoglobulin providing the CDRs is called the "donor," and the human immunoglobulin providing the framework is called the "receptor." In one embodiment, all CDRs are derived from donor immunoglobulins in the humanized immunoglobulin. The presence of a constant region is not essential, but if present, it must be substantially identical to the constant region of human immunoglobulin, i.e., at least about 85-90%, e.g., about 95% or more identical. Therefore, perhaps all parts of humanized immunoglobulin, except for the CDR, are substantially identical to the corresponding parts of the native human immunoglobulin sequence. A "humanized antibody" is an antibody containing humanized light chain and humanized heavy chain immunoglobulins. A humanized antibody binds to the same antigen as the donor antibody that provides the CDR. The receptor framework of a humanized immunoglobulin or antibody may have a limited number of substitutions by amino acids taken from the donor framework. A humanized antibody or other monoclonal antibody may have additional conserved amino acid substitutions that do not substantially affect antigen binding or other immunoglobulin functions. Exemplary conserved substitutions include those of gly, ala, val, ile, leu; asp, glu; asn, gln; ser, thr; lys, arg; and phe, tyr. Humanized immunoglobulins can be constructed by genetic engineering (see, for example, US5,585,089).Human antibodies are antibodies whose light and heavy chain genes are derived from humans. Human antibodies may be produced using methods known in the art. Human antibodies may also be produced by immortalizing human B cells that secrete the antibody of interest. Immortalization can be achieved, for example, by EBV infection, or by fusing human B cells with myeloma cells or fusion cell tumor cells to produce trioma cells. Human antibodies may also be produced by phage display (see, e.g., WO91 / 17271; WO92 / 001047; WO92 / 20791) or selected from a human combined monoclonal antibody library (see Morphosys website). Human antibodies may also be produced using genetically modified animals possessing human immunoglobulin genes (see, e.g., WO93 / 12227; WO91 / 10741).

[0118] Therefore, anti-ADM antibodies may have forms known in the art. Examples include human antibodies, monoclonal antibodies, humanized antibodies, chimeric antibodies, and CDR-transplanted antibodies. In preferred embodiments, the antibodies according to the present invention are antibody fragments comprising at least the F variable domain of a heavy chain and / or light chain, such as recombinantly produced antibodies like IgG, which is a typical full-length immunoglobulin, or chemically bound antibodies (fragment antigen binding), and these antibodies include, but are not limited to, Fab minibodies, single-chain Fab antibodies, monovalent Fab antibodies with epitope tags such as Fab-V5Sx2; bivalent Fab (mini-antibodies) dimerized with a CH3 domain; bivalent or polyvalent Fab formed via multimerization with the help of heterologous domains, such as dimerization of a dHLX domain such as Fab-dHLX-FSx2; F(ab')2 fragments, scFv fragments, multimerized polyvalent and / or polyspecific scFv fragments, bivalent and / or bispecific antibodies, BITE® (bispecific T cell engager), trifunctional antibodies, polyvalent antibodies; and single-domain antibodies, such as immunoglobulins from camelid or fish and antibodies from numerous other species.

[0119] In addition to anti-ADM antibodies, other biopolymer scaffolds that form complexes with target molecules are well known in the art and have been used to produce highly target-specific biopolymers. Examples include aptamers, spiegelmers, antikalins, and conotoxins. See Figures 1a, 1b, and 1c for descriptions of antibody forms.

[0120] In a preferred embodiment, the anti-ADM antibody form is selected from the group comprising Fv fragment, scFv fragment, Fab fragment, scFab fragment, F(ab)2 fragment, and scFv-Fc fusion protein. In another preferred embodiment, the antibody form is selected from the group comprising bioavailability-optimized complexes such as scFab fragment, Fab fragment, scFv fragment, and PEGylated fragment. One of the most preferred forms is the scFab form.

[0121] Non-Ig scaffolds may also be protein scaffolds, and since these scaffolds can bind to ligands or antigens, they may be used as antibody mimetic materials. Non-Ig scaffolds include tetranectin-based non-Ig scaffolds (e.g., disclosed in US2010 / 0028995), fibronectin scaffolds (e.g., disclosed in EP1 266 025); lipocalin-based scaffolds (e.g., disclosed in WO2011 / 154420); ubiquitin scaffolds (e.g., described in WO2011 / 073214); transferrin scaffolds (e.g., disclosed in US2004 / 0023334); protein A scaffolds (e.g., disclosed in EP2 231 860); ankyrin repeat-based scaffolds (e.g., disclosed in WO2010 / 060748); microprotein scaffolds, preferably microprotein scaffolds that form cysteine ​​knots (e.g., disclosed in EP2314308); Fyn The group may be selected from the SH3 domain-based scaffolding (e.g., disclosed in WO2011 / 023685), the EGFR-A domain-based scaffolding (e.g., disclosed in WO2005 / 040229), and the Knitz domain-based scaffolding (e.g., disclosed in EP1 941 867).

[0122] In one embodiment of the present invention, the anti-ADM antibody according to the present invention may be produced by synthesizing a fragment of ADM as an antigen, as outlined in Example 1. Subsequently, a binder to the fragment is identified using the method described below or other methods known in the art.

[0123] Humanization of mouse antibodies may be carried out according to the following procedure: For humanization of mouse-derived antibodies, the antibody sequence is analyzed for structural interactions between the complementarity region (CDR) and the antigen framework region (FR). Based on structural modeling, an appropriate human-derived FR is selected, and the mouse CDR sequence is transplanted into the human FR. Amino acid sequence mutations of the CDR or FR may be introduced to restore the structural interactions abolished by seed switching of the FR sequence. This restoration of structural interactions may be achieved by a random method using a phage display library or by a direct method induced by molecular modeling (Almagro and Fransson 2008. Front Biosci. 13: 1619-33).

[0124] In another embodiment, the anti-ADM antibody, anti-ADM antibody fragment, or anti-ADM non-Ig scaffold is a full-length antibody, antibody fragment, or non-Ig scaffold.

[0125] In one embodiment, an anti-ADM antibody or anti-ADM antibody fragment or anti-ADM non-Ig scaffold is directed to and can bind to an epitope of at least four or at least five amino acids in the N-terminal and / or central region (amino acids 1-42) of ADM-Gly and / or ADM-NH2 (SEQ ID NO: 23).

[0126] Epitopes, also known as antigenic groups, are parts of antigens (such as peptides and proteins) that are recognized by the immune system, particularly by antibodies. For example, an epitope is a specific part of an antigen to which an antibody binds. Some antibodies that bind to an epitope are called paratopes. Protein antigen epitopes are classified into two types based on their structure and interaction with paratopes: structural epitopes and linear epitopes.

[0127] Chain-like, or continuous, epitopes are epitopes recognized by antibodies due to their chain-like sequence of amino acids, i.e., their primary structure, and are formed by a three-dimensional structure adopted through the interaction of continuous amino acid residues. Structural epitopes and chain-like epitopes interact with paratopes based on the three-dimensional structure adopted by the epitope, and this structure is measured by the surface features of the epitope residues involved and the shape or tertiary structure of other compartments of the antigen. Structural epitopes are formed by a three-dimensional structure adopted through the interaction of discontinuous amino acid residues.

[0128] In a particular embodiment of the present invention, an anti-ADM antibody or anti-ADM antibody fragment or anti-ADM non-Ig scaffold can be directed to and bound to at least four or at least five amino acids in the N-terminal portion (amino acids 1-21) of ADM-Gly and / or ADM-NH2:YRQSMNNFQGLRSFGCRFGTC (SEQ ID NO: 14).

[0129] In another preferred embodiment, the anti-ADM antibody or anti-ADM antibody fragment or anti-ADM non-Ig scaffold is directed to and can bind to at least four or at least five amino acids in the N-terminal region (amino acids 1-14) of ADM-Gly and / or ADM-NH2 (SEQ ID NO: 25).

[0130] In another embodiment, the anti-ADM antibody or anti-ADM antibody fragment or anti-ADM non-Ig scaffold can be directed to and bound to at least four or at least five amino acids in the N-terminal region (amino acids 1-10):YRQSMNNFQG (SEQ ID NO: 26) of ADM-Gly and / or ADM-NH2.

[0131] In a very specific embodiment, the anti-ADM antibody or anti-ADM antibody fragment or anti-ADM non-Ig scaffold is directed to and can bind to at least four or at least five amino acids in the N-terminus (amino acids 1-6):YRQSMN (SEQ ID NO: 27) of ADM-Gly and / or ADM-NH2, and binding requires the free N-terminus (amino acid 1) of ADM and / or ADM-Gly.

[0132] In another, very specific embodiment of the present invention, an anti-ADM antibody or anti-adrenomedullin antibody fragment or anti-ADM non-Ig scaffold recognizes and binds to the N-terminus (amino acid 1) of ADM-Gly and / or ADM-NH2. The N-terminus means that amino acid 1, i.e., "Y" in SEQ ID NOs. 14, 20, 22, 23, 25, 26, and 27, is essential for antibody binding. This antibody or fragment or scaffold does not bind to N-terminally extended ADM, N-terminally modified ADM, or N-terminally degraded ADM-Gly and / or ADM-NH2. This means that when the N-terminus of ADM is free, the anti-ADM antibody or anti-ADM antibody fragment or non-Ig scaffold binds only to regions within the sequence of ADM-Gly and / or ADM-NH2. Anti-ADM antibodies, anti-ADM antibody fragments, or anti-ADM non-Ig scaffolds do not appear to bind to regions within the ADM-Gly and / or ADM-NH2 sequences if the sequence is included, for example, within pro-ADM.

[0133] For clarity, the numbers in parentheses in a specific region of ADM, such as "N-terminus (amino acids 1-21)," indicate that a person skilled in the art will understand that the N-terminus of ADM consists of amino acids 1-21 of the ADM-Gly and / or ADM-NH2 sequence.

[0134] In another specific embodiment of the present invention, the anti-ADM antibody or anti-ADM antibody fragment or anti-ADM non-Ig scaffold presented herein does not bind to the C-terminal portion of ADM, i.e., amino acids 43-52 (SEQ ID NO: 24) of ADM.

[0135] In a particular embodiment, it is preferable to use an anti-ADM antibody or anti-adrenomedullin antibody fragment or anti-ADM non-Ig scaffold according to the present invention, wherein the anti-adrenomedullin antibody or anti-adrenomedullin antibody fragment or non-Ig scaffold increases the ADM-NH2 level or ADM-NH2 immunoreactivity in serum, blood, or plasma by at least 10%, preferably at least 50%, more preferably more than 50%, and most preferably more than 100%.

[0136] A measurement method that can be used to measure the half-life (half-retention time) of adremedullin in serum, blood, and plasma is described in Example 3.

[0137] In certain embodiments of the present invention, the antibody is a monoclonal antibody or a fragment thereof. In one embodiment of the present invention, the anti-ADM antibody or anti-ADM antibody fragment is a human antibody or a humanized antibody, or derived therefrom. In one particular embodiment, one or more (mouse) CDRs are transplanted onto a human antibody or antibody fragment ("humanized").

[0138] In one aspect, the subject of the present invention is a humanized CDR implantation antibody or an antibody fragment thereof, the antibody recognizing or binding to the N-terminal portion of ADM-Gly and / or ADM-NH2 for treatment or interventional therapy in patients infected with coronavirus, and the humanized CDR implantation antibody or antibody fragment comprises an antibody heavy chain (H chain) comprising: Sequence ID 1: GYTFSRYW Sequence ID 2: ILPGSGST and / or Sequence ID 3: TEGYEYDGFDY and / or further comprising an antibody light chain (L chain) containing the following: Sequence ID 4: QSIVYSNGNTY Sequence "RVS" (not part of the sequence listing): RVS and / or Sequence ID 5: FQGSHIPYT.

[0139] A particular embodiment of the present invention is a humanized and / or human monoclonal antibody or antibody fragment thereof, wherein the antibody recognizes or binds to the N-terminal portion (amino acids 1-21) of ADM-Gly and / or ADM-NH2 (SEQ ID NO: 14) for the treatment or interventional therapy of patients infected with coronavirus, and the heavy chain comprises at least one CDR selected from the group including the following sequences: Sequence ID 1: GYTFSRYW Sequence ID 2: ILPGSGST and / or Sequence ID 3: TEGYEYDGFDY, Furthermore, the light chain contains at least one CDR selected from the group including the following sequences: Sequence ID 4: QSIVYSNGNTY Array "RVS" (not part of the array list): RVS Sequence ID 5: FQGSHIPYT.

[0140] In more specific embodiments of the present invention, the subject of the present invention is a humanized and / or human monoclonal antibody or an antibody fragment thereof, wherein the antibody recognizes or binds to the N-terminal portion (amino acids 1-21) of ADM-Gly and / or ADM-NH2 (SEQ ID NO: 14) for the treatment or interventional therapy of patients infected with coronavirus, and the heavy chain comprises the following sequence: Sequence ID 1: GYTFSRYW Sequence ID 2: ILPGSGST Sequence ID 3: TEGYEYDGFDY, Furthermore, the light chain contains the following sequence: Sequence ID 4: QSIVYSNGNTY Array "RVS" (not part of the array list): RVS Sequence ID 5 FQGSHIPYT.

[0141] In a very specific embodiment, the anti-ADM antibody has a sequence selected from the group including SEQ ID NOs: 6, 7, 8, 9, 10, 11, 12, 13, 35, and 36.

[0142] The anti-ADM antibody, anti-ADM antibody fragment, or anti-ADM non-Ig scaffold according to the present invention has an affinity constant of 10 -7 M or more, preferably 10 -8 M or more, more 10 -9 M or more, most preferably 10 -10 It exhibits affinity for human ADM-Gly and / or ADM-NH2 such that the affinity constant is greater than M. Those skilled in the art will know that administering higher doses of the compound may compensate for lower affinity, and that this measure is not outside the scope of the claims of the present invention. This affinity constant can be measured according to the method described in Example 1.

[0143] The subject of the present invention is a human or humanized monoclonal antibody or fragment that binds to ADM-Gly and / or ADM-NH2, wherein the antibody or fragment binds to the N-terminal portion (amino acids 1-21):YRQSMNNFQGLRSFGCRFGTC (SEQ ID NO: 14) of ADM-Gly and / or ADM-NH2 for the treatment or intervention of patients infected with coronavirus, and the antibody or fragment comprises a sequence selected from the group comprising the following sequences: Sequence number 6 (AM-VH-C): [ka] Sequence ID 7 (AM-VH1): [ka] Sequence ID 8 (AM-VH2-E40): [ka] Sequence ID 9 (AM-VH3-T26-E55): [ka] Sequence ID 10 (AM-VH4-T26-E40-E55): [ka] Sequence ID 11(AM-VL-C): [ka] Sequence ID 12 (AM-VL1): [ka] Sequence ID 13 (AM-VL2-E40): [ka] Sequence ID 35 (heavy chain HAM8101): [ka] Sequence ID 36 (Light Chain HAM 8101): [ka]

[0144] The subject of the present invention is further a human and / or humanized monoclonal antibody or fragment that binds to ADM-Gly and / or ADM-NH2, wherein the antibody or fragment binds to the N-terminal portion (amino acids 1-21):YRQSMNNFQGLRSFGCRFGTC (SEQ ID NO: 14) of ADM-Gly and / or ADM-NH2 for the treatment or intervention of patients infected with coronavirus, and the antibody or fragment comprises the following sequence as a heavy chain: Sequence ID 35: [ka] Furthermore, the light chain includes the following sequence: Sequence ID 36: [ka]

[0145] In a particular embodiment of the present invention, the antibody comprises the following sequence as a heavy chain: Sequence ID 35: [ka]

[0146] Alternatively, a sequence that is identical to that sequence by more than 95%, preferably more than 98%, preferably more than 99%, and includes the following sequence as a light chain: Sequence ID: 36 [ka] Alternatively, it is identical to that sequence by more than 95%, preferably more than 98%, and preferably more than 99%.

[0147] To assess the identity between two amino acid sequences, a pairwise sorting test is performed. Identity is defined as the percentage of amino acids that directly match within the sorting sequence.

[0148] The term "pharmaceutical preparation" means a pharmaceutical component combined with at least one pharmaceutically acceptable excipient, in a form that enables the bioactivity of the pharmaceutical component contained therein, and that does not contain additional components that are unacceptably toxic to the subject to which the preparation is administered. The term "pharmaceutical component" means a therapeutic composition that can be optionally combined with pharmaceutically acceptable excipients to provide a pharmaceutical preparation or dosage form.

[0149] The subject of the present invention is a pharmaceutical formulation comprising an antibody, fragment, or scaffold according to the present invention, for use in the treatment or interventional therapy of a patient infected with coronavirus.

[0150] The subject of the present invention is a pharmaceutical formulation used for the treatment or intervention therapy of patients infected with coronavirus according to the present invention, the pharmaceutical formulation being a solution, preferably a ready-to-use solution.

[0151] The subject of the present invention is a pharmaceutical formulation used for the treatment or intervention therapy of patients infected with coronavirus according to the present invention, the pharmaceutical formulation being in a lyophilized state.

[0152] The subject of the present invention is a pharmaceutical formulation used for the treatment or interventional therapy of a patient infected with the coronavirus according to the present invention, the pharmaceutical formulation being administered intramuscularly.

[0153] The subject of the present invention is a pharmaceutical formulation used for the treatment or interventional therapy of patients infected with coronavirus according to the present invention, the pharmaceutical formulation being administered intravascularly.

[0154] The subject of the present invention is a pharmaceutical formulation used for the treatment or intervention therapy of patients infected with coronavirus according to the present invention, the pharmaceutical formulation being administered by infusion.

[0155] The subject of the present invention is a pharmaceutical formulation for use in the treatment or intervention therapy of patients infected with coronavirus according to the present invention, said pharmaceutical formulation to be administered systemically. [Modes for carrying out the invention]

[0156] Embodiment 1. In patients infected with coronavirus, (a) methods for diagnosing or predicting the risk of life-threatening exacerbation or adverse events, (b) methods for diagnosing or determining the severity of the condition or prognosis, (c) methods for predicting or monitoring the success of treatment or interventional therapy, (d) methods for guiding or stratifying treatment, or (e) methods for managing patients, these methods are: - A step of measuring the level of proadrenomedullin (SEQ ID NO: 31) or its fragments in a body fluid sample from the patient. - A step of comparing the level of proadrenomedullin or its fragment with a predetermined threshold or a previously measured level of proadrenomedullin or its fragment, and - A process of correlating the level of proadrenomedullin or its fragments with the risk of life-threatening exacerbation or adverse events, or - A step of correlating the level of proadrenomedullin or its fragments with the severity, or The process of correlating the level of proadrenomedullin or its fragments with the success of a therapeutic or interventional treatment, The process of correlating the level of proadrenomedullin or its fragments with a specific treatment or intervention, or The process includes correlating the level of proadrenomedullin or its fragments with the management of the patient, The proadrenomedullin or its fragment is selected from the group consisting of PAMP (SEQ ID NO: 32), MR-proADM (SEQ ID NO: 33), ADM-NH2 (SEQ ID NO: 20), ADM-Gly (SEQ ID NO: 21), and CT-proADM (SEQ ID NO: 34).

[0157] 2. A method for (a) diagnosing or predicting the risk of life-threatening exacerbation or adverse events in a patient infected with the coronavirus described in Embodiment 1, or (b) determining the prognosis of the severity, or (c) predicting or monitoring the success of a treatment or intervention, wherein the coronavirus is selected from the group including Sars-CoV-1, Sars-CoV-2, and MERS-CoV, and is particularly Sars-CoV-2.

[0158] 3. A method for (a) diagnosing or predicting the risk of life-threatening exacerbation or adverse events in a patient infected with the coronavirus described in Embodiment 1 or 2, or (b) determining the prognosis of severity, or (c) predicting or monitoring the success of a treatment or intervention, wherein the adverse events are selected from a group including death, organ failure, and shock.

[0159] 4. In a patient infected with the coronavirus described in Embodiments 1 to 3, a method for (a) diagnosing or predicting the risk of life-threatening exacerbation or adverse events, or (b) determining the prognosis of severity, or (c) predicting or monitoring the success of a treatment or intervention, wherein the level of proadrenomedullin or a fragment thereof is above a predetermined threshold.

[0160] 5. A method for (a) diagnosing or predicting the risk of life-threatening exacerbation or adverse events in a patient infected with the coronavirus described in Embodiments 1 to 4, or (b) determining the prognosis of the severity, or (c) predicting or monitoring the success of a treatment or intervention, wherein the fragment is MR-proADM (SEQ ID NO: 33), and a predetermined threshold for MR-proADM in the subject's body fluid sample is 0.5 to 2 nmol / L, preferably 0.7 to 1.5 nmol / L, preferably 0.8 to 1.2 nmol / L, and most preferably a threshold of 1 nmol / L.

[0161] 6. A method for (a) diagnosing or predicting the risk of life-threatening exacerbation or adverse events in a patient infected with the coronavirus described in Embodiments 1 to 4, or (b) determining the prognosis of the severity, or (c) predicting or monitoring the success of a treatment or intervention, wherein the fragment is ADM-NH2 (SEQ ID NO: 20), and a predetermined threshold for ADM-NH2 (SEQ ID NO: 20) in the subject body fluid sample is 40 to 100 pg / mL, more preferably 50 to 90 pg / mL, even more preferably 60 to 80 pg / mL, and most preferably the threshold is 70 pg / mL.

[0162] 7. A method for (a) diagnosing or predicting the risk of life-threatening exacerbation or adverse events in a patient infected with the coronavirus described in Embodiments 1 to 6, or (b) determining the prognosis of the severity, or (c) predicting or monitoring the success of a treatment or intervention, wherein the patient's SOFA score is 3 or higher, preferably 7 or higher, or the patient's rapid SOFA score is 1 or higher, preferably 2 or higher.

[0163] 8. A method for (a) diagnosing or predicting the risk of life-threatening exacerbation or adverse events in a patient infected with the coronavirus described in Embodiments 1 to 7, or (b) determining the prognosis of the severity, or (c) predicting or monitoring the success of a treatment or intervention, wherein the patient has a D-dimer level of 0.5 μg / mL or higher, preferably 1.0 μg / mL or higher.

[0164] 9. A method for (a) diagnosing or predicting the risk of life-threatening exacerbation or adverse events in a patient infected with the coronavirus described in Embodiments 1 to 8, or (b) determining the prognosis of the severity, or (c) predicting or monitoring the success of a treatment or intervention, wherein the level of proadrenomedullin or its fragments is measured by contacting the body fluid sample with a capture agent that specifically binds to proadrenomedullin or its fragments.

[0165] 10. A method for (a) diagnosing or predicting the risk of life-threatening exacerbation or adverse events in a patient infected with the coronavirus described in Embodiments 1 to 9, or (b) determining the prognosis of severity, or (c) predicting or monitoring the success of a treatment or intervention, wherein the measurement comprises the use of a capture agent that specifically binds to proadrenomedullin or a fragment thereof, the capture agent being selected from the group consisting of antibodies, antibody fragments, or non-IgG scaffolds.

[0166] 11. A method for (a) diagnosing or predicting the risk of life-threatening exacerbation or adverse events in a patient infected with the coronavirus described in Embodiments 1 to 10, or (b) determining the prognosis of severity, or (c) predicting or monitoring the success of a treatment or intervention, comprising measuring the level of proadrenomedullin or a fragment thereof in a body fluid sample of the subject, wherein the measurement comprises the use of a capture binder that specifically binds to proadrenomedullin or a fragment thereof, the capture binder being an antibody.

[0167] 12. A method for (a) diagnosing or predicting the risk of life-threatening exacerbation or adverse events in a patient infected with the coronavirus described in Embodiments 1 to 11, or (b) determining the prognosis of severity, or (c) predicting or monitoring the success of a treatment or intervention, comprising measuring the level of proadrenomedullin or a fragment thereof in a body fluid sample of the subject, wherein the measurement includes the use of a scavenging agent that specifically binds to the level of proadrenomedullin or a fragment thereof, and the scavenging agent is immobilized on the surface.

[0168] 13. A method for (a) diagnosing or predicting the risk of life-threatening exacerbation or adverse events in a patient infected with the coronavirus described in Embodiments 1 to 12, or (b) determining the prognosis of the severity, or (c) predicting or monitoring the success of a treatment or intervention, wherein the patient is treated with an anti-adrenomedullin (ADM) antibody, an anti-ADM antibody fragment, or an anti-ADM non-Ig scaffold, wherein the anti-ADM antibody, anti-ADM fragment, or anti-ADM non-Ig scaffold is bound to the N-terminal and / or central region (amino acids 1 to 42) of ADM-Gly and / or ADM-NH2: YRQSMNNFQGLRSFGCRFGTCTVQKLAHQIYQFTDKDKDNVA (SEQ ID NO: 23).

[0169] 14. Anti-adrenomedullin (ADM) antibodies, anti-ADM antibody fragments, or anti-ADM non-Ig scaffolds for use in the treatment or intervention of patients infected with coronavirus.

[0170] 15. An anti-adrenomedullin (ADM) antibody or anti-ADM antibody fragment or anti-ADM non-Ig scaffold for use in the treatment or interventional therapy of a patient infected with the coronavirus described in Embodiment 14, wherein the coronavirus is selected from the group including Sars-CoV-1, Sars-CoV-2, and MERS-CoV, and is particularly Sars-CoV-2.

[0171] 16. An anti-adrenomedullin (ADM) antibody or anti-ADM antibody fragment or anti-ADM non-Ig scaffold for use in the treatment or interventional therapy of a patient infected with the coronavirus described in Embodiment 14 or 15, wherein the patient has a level of proadrenomedullin or its fragment in a body fluid sample of the subject that, when measured by the method of any one of claims 1 to 12, is above a predetermined threshold for proadrenomedullin or higher than a previously measured level.

[0172] 17. An anti-adrenomedullin (ADM) antibody, an anti-ADM antibody fragment, or an anti-ADM non-Ig scaffold for use in the treatment or interventional therapy of a patient infected with the coronavirus described in Embodiments 14 to 16, wherein the patient's SOFA score is 3 or higher, preferably 7 or higher, or the patient's rapid SOFA score is 1 or higher, preferably 2 or higher.

[0173] 18. An anti-adrenomedullin (ADM) antibody, an anti-ADM antibody fragment, or an anti-ADM non-Ig scaffold for use in the treatment or intervention therapy of a patient infected with the coronavirus described in Embodiments 14 to 17, wherein the patient has a D-dimer level of 0.5 μg / mL or higher, preferably 1.0 μg / mL or higher.

[0174] 19. An anti-adrenomedullin (ADM) antibody, an anti-ADM antibody fragment, or an anti-ADM non-Ig scaffold for use in the treatment or intervention therapy of a patient infected with the coronavirus described in Embodiments 14-18, wherein the anti-ADM antibody, anti-ADM fragment, or anti-ADM non-Ig scaffold is bound to the N-terminal portion (amino acids 1-21) of ADM-Gly and / or ADM-NH2: YRQSMNNFQGLRSFGCRFGTC (SEQ ID NO: 14).

[0175] 20. An anti-adrenomedullin (ADM) antibody, anti-ADM antibody fragment, or anti-ADM non-Ig scaffold for use in the treatment or intervention therapy of a patient infected with coronavirus as described in Embodiments 14-19, wherein the anti-adrenomedullin (ADM) antibody, anti-ADM antibody fragment, or anti-ADM non-Ig scaffold is 10% of the proadrenomedullin or its fragment. -7 It exhibits a minimum binding affinity of M or less.

[0176] 21. An anti-adrenomedullin (ADM) antibody, an anti-ADM antibody fragment, or an anti-ADM non-Ig scaffold used for the treatment or intervention therapy of a patient infected with the coronavirus described in Embodiments 14 to 20, wherein the anti-adrenomedullin (ADM) antibody, anti-ADM antibody fragment, or anti-ADM non-Ig scaffold inhibits the bioactivity of ADM by 80% or less, preferably 50% or less.

[0177] 22. An anti-adrenomedullin (ADM) antibody, an anti-ADM antibody fragment, or an anti-ADM non-Ig scaffold for use in the treatment or intervention therapy of a patient infected with the coronavirus described in Embodiments 14 to 21, wherein the antibody is a monoclonal antibody or a monoclonal antibody fragment.

[0178] 23. An anti-adrenomedullin (ADM) antibody or anti-ADM antibody fragment or anti-ADM non-Ig scaffold for use in the treatment or intervention therapy of a patient infected with coronavirus as described in Embodiment 22, wherein the complementarity-determining region (CDR) within the heavy chain comprises the following sequence: CDR1: Sequence ID 1 GYTFSRYW CDR2: Sequence ID 2 ILPGSGST CDR3: Sequence ID 3 TEGYEYDGFDY Furthermore, the complementarity-determining region (CDR) within the light chain contains the following sequence: CDR1: Sequence ID 4 QSIVYSGNTY CDR2: RVS CDR3: Sequence ID 5 FQGSHIPYT.

[0179] 24. An anti-adrenomedullin (ADM) antibody, an anti-ADM antibody fragment, or an anti-ADM non-Ig scaffold for use in the treatment or intervention therapy of a patient infected with the coronavirus described in Embodiment 23, wherein the antibody or fragment is a sequence selected from the group comprising the following sequences as the VH region: Sequence number 6 (AM-VH-C): [ka] Sequence ID 7 (AM-VH1): [ka] Sequence ID 8 (AM-VH2-E40): [ka] Sequence ID 9 (AM-VH3-T26-E55): [ka] Sequence ID 10 (AM-VH4-T26-E40-E55): [ka] Or it includes sequences that are more than 80% identical to each of the above sequences, and Sequences selected from the group containing the following sequences as the VL region: Sequence ID 11(AM-VL-C): [ka] Sequence ID 12 (AM-VL1): [ka] Sequence ID 13 (AM-VL2-E40): [Chemical formula] or includes a sequence that is more than 80% identical to each of the above sequences.

[0180] 25. An anti - adrenomedullin (ADM) antibody or anti - ADM antibody fragment or anti - ADM non - Ig scaffold for use in the treatment or interventional treatment of a patient infected with the coronavirus according to any one of embodiments 23 to 24, wherein the antibody or fragment has the following sequence as the heavy chain: SEQ ID NO: 35: [Chemical formula] or includes a sequence that is more than 95% identical thereto, and has the following sequence as the light chain: SEQ ID NO: 36: [Chemical formula] or includes a sequence that is more than 95% identical thereto.

[0181] 26. An anti - adrenomedullin (ADM) antibody or anti - ADM antibody fragment or anti - ADM non - Ig scaffold for use in the treatment or interventional treatment of a patient infected with the coronavirus according to any one of embodiments 23 to 25, wherein the monoclonal antibody or antibody fragment is a humanized monoclonal antibody or humanized monoclonal antibody fragment.

[0182] 27. An anti - adrenomedullin (ADM) antibody or anti - ADM antibody fragment or anti - ADM non - Ig scaffold for use in the treatment or interventional treatment of a patient infected with the coronavirus according to any one of embodiments 14 to 26, wherein the anti - adrenomedullin (ADM) antibody or anti - ADM antibody fragment or anti - ADM non - Ig scaffold is a monoclonal antibody and is adrecizumab, and has the following sequence as the heavy chain: SEQ ID NO: 35: <00​​ and as the light chain the following sequence: SEQ ID NO: 36:

Chemical Formula

[0183] 28. An anti - adrenomedullin (ADM) antibody or an anti - ADM antibody fragment or an anti - ADM non - Ig scaffold for use in the treatment or interventional treatment of a patient suffering from pulmonary dysfunction and / or acute respiratory distress syndrome (ARDS).

[0184] 29. An anti - adrenomedullin (ADM) antibody or an anti - ADM antibody fragment or an anti - ADM non - Ig scaffold for use in the treatment or interventional treatment of a patient suffering from pulmonary dysfunction and / or acute respiratory distress syndrome (ARDS) according to embodiment 28, wherein the patient has a Holowitz index of less than 300, particularly less than 200, particularly less than 100, and / or the patient requires a ventilator.

[0185] 30. An anti - adrenomedullin (ADM) antibody or an anti - ADM antibody fragment or an anti - ADM non - Ig scaffold for use in the treatment or interventional treatment of a patient suffering from pulmonary dysfunction and / or acute respiratory distress syndrome (ARDS) according to embodiment 28 or 29, wherein the patient has a level of pro - adrenomedullin or a fragment thereof in the body fluid sample of the subject that exceeds a predetermined threshold of pro - adrenomedullin or is higher than a previously measured level when measured by the method according to any one of claims 1 - 12.

[0186] 31. An anti - adrenomedullin (ADM) antibody or an anti - ADM antibody fragment or an anti - ADM non - Ig scaffold for use in the treatment or interventional treatment of a patient suffering from pulmonary dysfunction and / or acute respiratory distress syndrome (ARDS) according to embodiments 28 - 30, wherein the SOFA score of the patient is 3 or more, preferably 7 or more, or the quick SOFA score of the patient is 1 or more, preferably 2 or more.

[0187] 32. An anti-adrenomedullin (ADM) antibody, an anti-ADM antibody fragment, or an anti-ADM non-Ig scaffold for use in the treatment or intervention of a patient suffering from pulmonary dysfunction and / or acute respiratory spurt syndrome (ARDS) as described in Embodiments 28 to 31, wherein the patient has a D-dimer level of 0.5 μg / mL or higher, preferably 1.0 μg / mL or higher.

[0188] 33. An anti-adrenomedullin (ADM) antibody, an anti-ADM antibody fragment, or an anti-ADM non-Ig scaffold for use in the treatment or intervention of patients suffering from pulmonary dysfunction and / or acute respiratory spurt syndrome (ARDS) as described in Embodiments 28-32, wherein the anti-ADM antibody, anti-ADM fragment, or anti-ADM non-Ig scaffold is bound to the N-terminal portion (amino acids 1-21):YRQSMNNFQGLRSFGCRFGTC (SEQ ID NO: 14) of ADM-Gly and / or ADM-NH2.

[0189] 34. An anti-adrenomedullin (ADM) antibody, anti-ADM antibody fragment, or anti-ADM non-Ig scaffold for use in the treatment or intervention of a patient suffering from pulmonary dysfunction and / or acute respiratory urgency syndrome (ARDS) as described in Embodiments 28-33, wherein the anti-adrenomedullin (ADM) antibody, anti-ADM antibody fragment, or anti-ADM non-Ig scaffold is 10% of the proadrenomedullin or its fragment. -7 It exhibits a minimum binding affinity of M or less.

[0190] 35. An anti-adrenomedullin (ADM) antibody, an anti-ADM antibody fragment, or an anti-ADM non-Ig scaffold used for the treatment or intervention of patients suffering from pulmonary dysfunction and / or acute respiratory spurt syndrome (ARDS) as described in Embodiments 28 to 34, wherein the anti-adrenomedullin (ADM) antibody, anti-ADM antibody fragment, or anti-ADM non-Ig scaffold inhibits the bioactivity of ADM by 80% or less, preferably 50% or less.

[0191] 36. An anti - adrenomedullin (ADM) antibody or anti - ADM antibody fragment or anti - ADM non - Ig scaffold for use in the treatment or interventional treatment of a patient suffering from the pulmonary function disorder and / or acute respiratory distress syndrome (ARDS) according to Embodiments 28 to 35, wherein the antibody is a monoclonal antibody or monoclonal antibody fragment.

[0192] 37. An anti - adrenomedullin (ADM) antibody or anti - ADM antibody fragment or anti - ADM non - Ig scaffold for use in the treatment or interventional treatment of a patient suffering from the pulmonary function disorder and / or acute respiratory distress syndrome (ARDS) according to Embodiment 36, wherein the complementarity - determining region (CDR) in the heavy chain comprises the following sequences: CDR1: SEQ ID NO: 1 GYTFSRYW CDR2: SEQ ID NO: 2 ILPGSGST CDR3: SEQ ID NO: 3 TEGYEYDGFDY And the complementarity - determining region (CDR) in the light chain comprises the following sequences: CDR1: SEQ ID NO: 4 QSIVYSNGNTY CDR2: RVS CDR3: SEQ ID NO: 5 FQGSHIPYT.

[0193] 38. An anti - adrenomedullin (ADM) antibody or anti - ADM antibody fragment or anti - ADM non - Ig scaffold for use in the treatment or interventional treatment of a patient suffering from the pulmonary function disorder and / or acute respiratory distress syndrome (ARDS) according to Embodiment 37, wherein the antibody or fragment comprises a sequence selected from the group consisting of the following sequences as the VH region: SEQ ID NO: 6 (AM - VH - C):

Chemical formula

Chemical formula

[0194] 39. An anti-adrenomedullin (ADM) antibody or anti-ADM antibody fragment or anti-ADM non-Ig scaffold for use in the treatment or intervention of a patient suffering from pulmonary dysfunction and / or acute respiratory urgency syndrome (ARDS) as described in any of embodiments 37 to 38, wherein the antibody or fragment has the following sequence as its heavy chain: Sequence ID 35: [ka] or containing sequences that are more than 95% identical to it, Furthermore, the light chain has the following sequence: Sequence ID 36: [ka] Or it includes sequences that are more than 95% identical to it.

[0195] 40. An anti-adrenomedullin (ADM) antibody or anti-ADM antibody fragment or anti-ADM non-Ig scaffold for use in the treatment or intervention of a patient suffering from pulmonary dysfunction and / or acute respiratory urgency syndrome (ARDS) as described in any of embodiments 37 to 39, wherein the monoclonal antibody or antibody fragment is a humanized monoclonal antibody or humanized monoclonal antibody fragment.

[0196] 41. An anti-adrenomedullin (ADM) antibody, anti-ADM antibody fragment, or anti-ADM non-Ig scaffold for use in the treatment or intervention of patients suffering from pulmonary dysfunction and / or acute respiratory urgency syndrome (ARDS) as described in Embodiments 28-40, wherein the anti-adrenomedullin (ADM) antibody, anti-ADM antibody fragment, or anti-ADM non-Ig scaffold is a monoclonal antibody and is adrecizumab, and has the following sequence as its heavy chain: Sequence ID 35: [ka] And the following sequence as the light chain: Sequence ID 36: [ka] It contains or is a biosimilar thereof. [Examples]

[0197] Examples It is important to emphasize that, in accordance with the present invention, some of the antibodies, antibody fragments, and non-Ig scaffolds in the examples bind to ADM, and therefore should be considered as anti-ADM antibodies / antibody fragments / non-Ig scaffolds.

[0198] Example 1 - Antibody Production and Measurement of its Affinity Constant Several anti-human and anti-mouse ADM antibodies were generated, and their affinity constants were measured (see Tables 2 and 3).

[0199] Immunotherapy peptides / complexes: As shown in Table 2, (if cysteine ​​was not present in the selected ADM sequence) an immunization peptide (JPT Technologies, Berlin, Germany) was synthesized using an additional N-terminal cysteine ​​residue to bind the peptide to bovine serum albumin (BSA). The peptide was covalently bound to BSA using a Sulfolink-binding gel (Perbio-science, Bonn, Germany). The binding procedure was carried out according to Perbio's instructions.

[0200] Mouse monoclonal antibody production: Balb / c mice were immunized with 100 μg of peptide-BSA conjugate (emulsified in 100 μL of complete Freund's retrieval) on days 0 and 14, and with 50 μg of the same conjugate (emulsified in 100 μL of incomplete Freund's retrieval) on days 21 and 28. Three days before performing the fusion test, the animals were administered 50 μg of the conjugate dissolved in 100 μL of saline via a single intraperitoneal injection and a single intravenous injection. Splenocytes and myeloma cell lineage SP2 / 0 cells from the immunized mice were fused with 1 mL of 50% polyethylene glycol at 37°C for 30 seconds. After washing, these cells were seeded in 96-well cell culture plates. Hybrid clones were selected by growing them in HAT medium [RPMI1640 culture medium supplemented with 20% fetal bovine serum and HAT supplement]. After two weeks, the HAT medium was replaced with HT medium and the cells were subcultured three times, after which the cells were returned to normal cell culture medium.

[0201] Cell culture supernatants were primary screened for antigen-specific IgG antibodies three weeks after fusion. Tested positive microcultures were transferred to 24-well plates for growth. After retesting, selected cultures were cloned and re-cloned using limiting dilution to determine isotype (see Lane, RD 1985. J. Immunol. Meth. 81: 223-228; Ziegler et al. 1996. Horm. Metab. Res. 28: 11-15).

[0202] Antibodies were produced using a standard antibody production method (Marx et al, 1997. Monoclonal Antibody Production, ATLA 25, 121) and purified with Protein A. Antibody purity was over 95% based on SDS gel electrophoresis analysis.

[0203] Human antibody production via phage display: The human naive antibody gene library HAL7 / 8 was used to isolate recombinant single-chain F variable domains (scFv) against the adrenomedullin peptide. The antibody gene library was sorted using a panning method that included the use of peptides containing biotin tags linked to the adrenomedullin peptide sequence via two different spacers. Background from nonspecific binders was minimized using a combination of panning cycles with nonspecific binding antigens and streptavidin-binding antigens. Phages eluted from the third panning cycle were used to generate monoclonal scFv expressing the E. coli strain. The supernatant from the cultures of these clones was used directly for antigen ELISA (see Hust et al. 2011. Journal of Biotechnology 152, 159-170; Schutte et al. 2009. PLoS One 4, e6625).

[0204] Positive clones were selected based on positive ELISA signals to the antigen and negative ELISA signals to streptavidin-coated microtiter plates. For further characterization, scFv open reading frames were cloned into the expression plasmid pOPE107 (Hust et al., J. Biotechn. 2011), captured from the culture supernatant by fixed metal ion affinity chromatography, and purified by size exclusion chromatography.

[0205] affinity constant To measure the affinity of antibodies to adrenomedullin, the binding dynamics of adrenomedullin to immobilized antibodies were measured by label-free surface plasmon resonance using a Biacore 2000 system (GE Healthcare Europe GmbH, Freiburg, Germany). Reversible antibody immobilization was performed using anti-mouse Fc antibodies covalently bound to the CM5 sensor surface at high density, according to the manufacturer's instructions (mouse antibody capture kit; GE Healthcare). (Lorenz et al. 2011. Antimicrob Agents Chemother. 55(1): 165-173).

[0206] Monoclonal antibodies were grown against the following ADM regions of human ADM and mouse ADM, respectively. The table below shows the selection of isolated antibodies for further testing. The selection is based on the target region. [Table 2] The following is a list of the monoclonal antibodies that were further collected. [Table 3-1] [Table 3-2]

[0207] Production of antibody fragments by enzymatic digestion: Fab and F(ab)2 fragments were generated by enzymatic digestion of the full-length mouse antibody NT-M. Antibody NT-M was digested using a) a pepsin-based F(ab)2 preparation kit (Pierce 44988) and b) a papain-based Fab preparation kit (Pierce 44985). The fragmentation procedure was carried out according to the instructions provided by the supplier. For F(ab)2 fragmentation, digestion was carried out at 37°C for 8 hours, and for Fab fragmentation, digestion was carried out for 16 hours.

[0208] Procedure for generating and purifying Fab: The fixed papain was washed with 0.5 mL of digestion buffer, and the column was equilibrated by centrifugation at 5000 x g for 1 minute. The buffer was then discarded. A desalted column was prepared by removing the storage solution, washing with digestion buffer, and then centrifugation at 1000 x g for 2 minutes each time. 0.5 mL of the prepared IgG sample was added to a spin column tube containing equilibrated fixed papain. The culture time for the digestion reaction was 16 hours at 37°C on a benchtop oscillating apparatus. This column was centrifuged at 5000 x g for 1 minute to separate the digest from the fixed papain. The resin was then washed with 0.5 mL of PBS and centrifuged at 5000 x g for 1 minute. The washed fraction was added to the digested antibody to make a total sample volume of 1.0 mL. The Nab protein A column was equilibrated at room temperature with PBS and IgG elution buffer. This column was centrifuged for 1 minute to remove the storage solution (containing 0.02% sodium azide), 2 mL of PBS was added to equilibrate, and the column was centrifuged again for 1 minute to discard the pass-through fraction. The sample was applied to the column and resuspended by inversion. The culture was performed at room temperature for 10 minutes by inversion mixing. The column was centrifuged for 1 minute, and the circulating fraction was stored together with the Fab fragment. (References: Coulter and Harris 1983. J. Immunol. Meth. 59, 199-203.;Lindner et al. 2010. Cancer Res. 70, 277-87;Kaufmann et al. 2010. PNAS. 107, 18950-5.;Chen et al. 2010. PNAS. 107, 14727-32; Uysal et al. 2009 J. Exp. Med. 206, 449-62; Thomas et al. 2009. J. Exp. Med. 206, 1913-27; Kong et al. 2009 J. Cell Biol. 185, 1275-840).

[0209] Procedure for generating and purifying the F(ab')2 fragment: The fixed pepsin was washed with 0.5 mL of digestion buffer, and the column was equilibrated by centrifugation at 5000 x g for 1 minute. The buffer was then discarded. Desalting columns were prepared by removing the storage solution, washing with digestion buffer, and then centrifugation at 1000 x g for 2 minutes each time. 0.5 mL of the prepared IgG sample was added to a spin column tube containing equilibrated fixed pepsin. The digestion reaction culture time was 16 hours at 37°C on a benchtop shaking apparatus. This column was centrifuged at 5000 x g for 1 minute to separate the digest from the fixed papain. The resin was then washed with 0.5 mL of PBS and centrifuged at 5000 x g for 1 minute. The washed fraction was added to the digested antibody to make a total sample volume of 1.0 mL. The Nab protein A column was equilibrated at room temperature with PBS and IgG elution buffer. The column was centrifuged for 1 minute to remove the storage solution (containing 0.02% sodium azide), 2 mL of PBS was added to equilibrate, and the column was centrifuged again for 1 minute to discard the passed fraction. The sample was applied to the column and resuspended by inversion. The culture was carried out at room temperature for 10 minutes by inversion mixing. The column was centrifuged for 1 minute, and the passed fraction was stored together with the Fab fragment.(References:Mariani et al. 1991. Mol. Immunol. 28: 69-77;Beale 1987. Exp Comp Immunol 11: 287-96;Ellerson et al. 1972. FEBS Letters 24(3): 318-22;Kerbel and Elliot 1983. Meth Enzymol 93: 113-147;Kulkarni et al. 1985. Cancer Immunol Immunotherapy 19:211-4;Lamoyi 1986. Meth Enzymol 121: 652-663;Parham et al. 1982. J Immunol Meth 53: 133-73;Raychaudhuri et al. 1985. Mol Immunol 22(9): 1009-19;Rousseaux et al. 1980. Mol Immunol 17: 469-82;Rousseaux et al. 1983. J Immunol Meth 64: 141-6;Wilson et al. 1991. J Immunol Meth 138: 111-9).

[0210] Humanization of NT-H antibody fragments: Antibody fragments were humanized by CDR transplantation (Jones et al. 1986. Nature 321, 522-525). To achieve the humanized sequence, the following steps were performed: Total RNA was extracted from NT-H fusion cell tumors using the Qiagen kit. For the first RT-PCR, the QIAGEN® one-step RT-PCR kit (catalog number 210210) was used. RT-PCR was performed using heavy and light chain specific primer sets. For each RNA sample, RT-PCR reactions for 12 heavy chains and 11 light chains were set up using a degenerate forward primer mixture containing variable region leader sequences. Reverse primers were positioned within the constant regions of the heavy and light chains. Restriction sites were not designed within the primers.

[0211] The reaction setup was as follows: 5.0 μL of 5x QIAGEN® one-step RT-PCR buffer, 0.8 μL of dNTP mixture (containing 10 mM of each dNTP), 0.5 μL of primer set, 0.8 μL of QIAGEN® one-step RT-PCR enzyme mixture, 2.0 μL of template RNA, and 20.0 μL of RNA-degrading enzyme-free solution, for a total volume of 20.0 μL. The PCR conditions were: reverse transcription at 50°C for 30 minutes; initial PCR activation at 95°C for 15 minutes; 20 cycles at 94°C for 25 seconds; 54°C for 30 seconds; 72°C for 30 seconds; and final extension at 72°C for 10 minutes. Second semi-nested PCR: The RT-PCR product from the first reaction was further amplified in the second PCR. RT-PCR reactions for 12 heavy chains and 11 light chains were set up using semi-nested primer sets specific to the antibody variable region.

[0212] The reaction setup was as follows: 10 μL of double PCR mixture; 2 μL of primer set; and a total volume of 20 μL including 8 μL of the first PCR product. The PCR conditions for the antibody cloning report of fusion cell tumor were: initial denaturation at 95°C for 5 minutes; 25 cycles at 95°C for 25 seconds; 57°C for 30 seconds; 68°C for 30 seconds; and final extension at 68°C for 10 minutes.

[0213] After PCR was completed, the PCR reaction sample was run on an agarose gel to visualize the amplified DNA fragments. Following sequencing of more than 15 cloned DNA fragments amplified by nested RT-PCR, the heavy and light chains of several mouse antibodies were cloned and accurately displayed. Protein sequence alignment and CDR analysis identified one heavy chain and one light chain. After alignment to a homologous human framework sequence, the resulting humanized sequence of the variable heavy chain is shown in Figure 5. The amino acids at positions 26, 40, and 55 in the variable heavy chain and the amino acid at position 40 in the variable light chain are important for binding properties, and these amino acids may be returned to the original mouse sequence. The resulting candidates are shown below. (Padlan 1991. Mol. Immunol. 28: 489-498; Harris and Bajorath 1995. Protein Sci. 4: 306-310).

[0214] For reference, the antibody fragment sequences (SEQ ID NOs. 7-13, 35, and 36) are shown below, with the bolded and underlined sequences being CDR1, 2, and 3, respectively, arranged over time. Sequence number 6 (AM-VH-C) [ka] Sequence ID 7 (AM-VH1) [ka] Sequence ID 8 (AM-VH2-E40) [ka] Sequence ID 9 (AM-VH3-T26-E55) [ka] Sequence ID 10 (AM-VH4-T26-E40-E55) [ka] Sequence ID 11 (AM-VL-C) [ka] Sequence ID 12 (AM-VL1) [ka] Sequence ID 13 (AM-VL2-E40) [ka] Sequence ID 35 [ka] Sequence ID 36 [ka]

[0215] Example 2 - Effect of selected anti-ADM antibodies on anti-ADM bioactivity The effect of selected ADM antibodies on ADM bioactivity was tested using the human recombinant adrenomedullin receptor cAMP function assay (adrenomedullin bioassay). The following materials were used: cell line CHO-K1, adrenomedullin receptor (CRLR+RAMP3); receptor cell line accession numbers (CRLR:U17473; RAMP3:AJ001016). CHO-K1 cells expressing human recombinant adrenomedullin receptor (FAST-027C), grown in antibiotic-free medium prior to the test, were separated by slow flow with PBS-EDTA (5 mM EDTA), recovered by centrifugation, and resuspended in the measurement buffer (KRH: 5 mM KCl, 1.25 mM MgSO4, 124 mM NaCl, 25 mM HEPES, 13.3 mM glucose, 1.25 mM KH2PO4, 1.45 mM CaCl2, 0.5 g / L BSA). Dose-response curves were measured together with a control agent (hADM or mADM).

[0216] Antagonist test (96 wells): For antagonist testing, 6 μL of control agonist (human (5.63 nM) or mouse (0.67 nM) adrenomedullin) was mixed with 6 μL of test sample or 6 μL of buffer in various antagonist dilutions. After incubation at room temperature for 60 minutes, 12 μL of cells (2,500 cells / well) were added. The plates were incubated at room temperature for 30 minutes. After adding lysis buffer, the DeltaF percentage was estimated using the Cis-Bio International HTRF kit (catalog number: 62AM2 PEB) according to the manufacturer's specifications. hADM22-52 were used as control antagonists. Antibodies used to test the cAMP-HTRF assay:

[0217] Anti-h-ADM antibodies (NT-H, MR-H, CT-H) were tested for antagonist activity in human recombinant adrenomedullin receptor (FAST-027C) cAMP function measurement in the presence of 5.63 nM human ADM1-52 (SEQ ID NO: 20) at final antibody concentrations of 100 μg / mL, 20 μg / mL, 4 μg / mL, 0.8 μg / mL, and 0.16 μg / mL. Anti-m-ADM antibodies (NT-M, MR-M, CT-M) were tested for antagonist activity in human recombinant adrenomedullin receptor (FAST-027C) cAMP function measurement in the presence of 0.67 nM mouse ADM1-50 (SEQ ID NO: 22) at final antibody concentrations of 100 μg / mL, 20 μg / mL, 4 μg / mL, 0.8 μg / mL, and 0.16 μg / mL. The data plots the relative inhibition rate against the antagonist concentration (see Figures 2a-2l). The maximum inhibition rates for each antibody are shown in Table 4. [Table 4]

[0218] Example 3 - Stabilization of hADM with anti-ADM antibody The stabilizing effect of human ADM antibodies on human ADM was tested using hADM immunoassay. The technique used was a sandwich tube luminescence immunoassay based on acridinium ester labeling.

[0219] Labeled compound (tracer): 100 μg (100 μL) of CT-H (1 mg / mL in PBS, pH 7.4, AdrenoMed AG Germany) was mixed with 10 μL of acridinium NHS ester (1 mg / mL in acetonitrile, InVent GmbH, Germany) (EP0353971) and incubated at room temperature for 20 minutes. Labeled CT-H was purified by gel filtration HPLC using Bio-Sil® SEC400-5 (Bio-Rad Laboratories, Inc., USA). The purified CT-H was diluted in (300 mmol / L potassium phosphate, 100 mmol / L NaCl, 10 mmol / L Na-EDTA, 5 g / L bovine serum albumin, pH 7.0). The final concentration was approximately 800,000 relative light units (RLU) per 200 μL of labeled compound (approximately 20 ng of labeled antibody). The chemiluminescence of the acridinium ester was measured using AutoLumat LB 953 (Berthold Technologies GmbH & Co. KG).

[0220] Solid phase: A polystyrene tube (Greiner Bio-One International AG, Austria) was coated with MR-H (AdrenoMed AG, Germany) (1.5 μg of MR-H / 0.3 mL with 100 mmol / L of NaCl, 50 mmol / L of TRIS / HCl, pH 7.8) for 18 hours at room temperature. After shielding with 5% bovine serum albumin, the tube was washed with PBS, pH 7.4, and vacuum-dried.

[0221] Calibration: This measurement method was calibrated using hADM (BACHEM AG, Switzerland) diluted in 250 mmol / L NaCl, 2 g / L Triton X-100, 50 g / L bovine serum albumin, and a 20 tab / L mixture of protein enzyme inhibitors (Roche Diagnostics AG, Switzerland).

[0222] hADM immunoassay: 50 μL of sample (or calibrator) was pipetteed into a coated tube, labeled CT-H (200 μL) was added, and the tube was incubated at 4°C for 4 hours. Unbound tracers were removed by washing five times (1 mL each) with a washing solution (20 mM PBS, pH 7.4, 0.1% Triton X-100). Chemiluminescence bound to the tube was measured using LB 953 (Berthold, Germany). Figure 3 shows a typical hADM dose / signal curve and the hADM dose / signal curve in the presence of 100 μg / mL NT-H antibody. NT-H did not affect the hADM immunoassay described above.

[0223] Stability of human adrenomedullin: Human adrenomedullin (HDM) was diluted in human citrate plasma (final concentration 10 nM) and cultured at 24°C. At predetermined time points, the degradation of hADM was stopped by freezing at -20°C. This culture was performed both with and without NT-H (100 μg / mL). The residual hADM was quantified using the hADM immunoassay method described above. Figure 4 shows the stability of hADM in human plasma (citrate) in the absence and presence of NT-H antibody. The half-life of hADM alone was 7.8 hours, but in the presence of NT-H, the half-life was 18.3 hours (2.3 times higher stability).

[0224] Example 4 - Mortality of sepsis a) Early treatment of sepsis Animal model: Male C57Bl / 6 mice (Charles River Laboratories, Germany) aged 12-15 weeks were used for the study. Peritonitis was surgically induced under mild isoflurane anesthesia. An incision was made in the upper left quadrant of the abdominal cavity (the normal location of the cecum). The cecum was exposed, and the area around the cecum distal to the small intestine insertion site was tightly ligated with sutures. A single hole was made in the cecum with a 24-gauge needle, and a small amount of cecal contents were squeezed out through the hole. The cecum was returned to the abdominal cavity, and the abdominal opening was closed. Finally, the animals were returned to their cages, and food and water were allowed to be freely consumed. 500 μL of physiological saline was administered subcutaneously as replacement fluid.

[0225] Administration and dosage of compounds (NT-M, MR-M, CT-M): Mice were treated immediately after CLP (early treatment). CLP stands for cecal ligation and puncture.

[0226] Test Groups: Three compounds were tested against treatment with the medium and a control compound. Each group consisted of 5 mice, with blood collected 1 day later for BUN (serum blood urea nitrogen test) measurement. Ten additional mice from each group were followed for 4 days. Therapeutic dose (10 μL / g body weight) in each group / Follow-up: 1: 4-day survival rate of NT-M, 0.2 mg / mL 2: 4-day survival rate for MR-M, 0.2 mg / mL 3: 4-day survival rate of CT-M, 0.2 mg / mL 4: Survival rate of nonspecific mouse IgG at 0.2 mg / mL over 4 days 5: Control - Survival rate over 4 days with PBS, 10 μL / g body weight

[0227] Clinical Chemistry: Blood urea nitrogen (BUN) concentrations for renal function were measured as a baseline and on day 1 after CLP. Blood samples were collected from the cavernous sinus using capillaries under mild ether anesthesia. Measurements were performed using an AU 400 Olympus Multianalyser. Mortality and mean BUN concentrations over 4 days are shown in Table 5. [Table 5]

[0228] Table 4 shows that NT-M antibodies significantly reduced mortality. 70% of mice treated with NT-M antibodies survived after 4 days. 30% of mice treated with MR-M antibodies survived, and 10% of mice treated with CT-M antibodies survived after 4 days. In contrast, all mice treated with nonspecific mouse IgG died after 4 days. Similar results were obtained in the control group administered PBS (phosphate-buffered saline). Blood urea nitrogen (BUN) testing was used to assess renal function, aid in the diagnosis of renal disease, and monitor patients with acute or chronic renal impairment or failure. The results of the S-BUN test clearly demonstrated that NT-M antibodies were most effective in protecting the kidneys.

[0229] b) Late-stage treatment of sepsis Animal model: Male C57Bl / 6 mice (Charles River Laboratories, Germany) aged 12-15 weeks were used in this study. Peritonitis was surgically induced under mild isoflurane anesthesia. An incision was made in the upper left quadrant of the abdominal cavity (the normal location of the cecum). The cecum was exposed, and the area around the cecum distal to the small intestine insertion site was tightly ligated with sutures. A single hole was made in the cecum with a 24-gauge needle, and a small amount of cecal contents were squeezed out through this hole. The cecum was returned to the abdominal cavity, and the abdominal incision was closed. Finally, the animals were returned to their cages, and food and water were allowed to be freely consumed. 500 μL of physiological saline was administered subcutaneously as replacement fluid.

[0230] Dosage and administration of the compound (NT-M FAB2): NT-M FAB2 was tested against treatment with the medium and control compounds. Treatment was administered 6 hours after CLP (late treatment) after sepsis had fully developed. Each group consisted of 4 mice, which were followed for 4 days. Therapeutic dose (10 μL / g body weight) in each group / Follow-up: 1. 4-day survival rate of NT-M FAB2, 0.2 mg / mL 2. Survival rate over 4 days of reference nonspecific mouse IgG, 0.2 mg / mL 3. Survival rate over 4 days using PBS at 10 μL / g body weight. [Table 6]

[0231] Table 6 shows that the NT-M FAB2 antibody significantly reduced mortality. 75% of mice treated with the NT-M FAB2 antibody survived after 4 days. In contrast, all mice treated with nonspecific mouse IgG died after 4 days. Similar results were obtained in the control group administered PBS (phosphate-buffered saline).

[0232] Example 5 - Administration of NT-H to healthy humans The study was conducted as a randomized, double-blind, placebo-controlled study in healthy men. A single escalating dose of NT-H antibody was administered intravenously (iv) to three consecutive groups of eight healthy men. These groups corresponded to the respective doses given to healthy men (six receiving the active drug and two receiving a placebo) (Group 1: 0.5 mg / kg, Group 2: 2 mg / kg, Group 3: 8 mg / kg). The main selection criteria were informed consent, i.e., age 18-35 years, agreement to use reliable contraception, and a BMI of 18-30 kg / m². 2 The study involved a single intravenous injection of either NT-H antibody (0.5 mg / kg; 2 mg / kg; 8 mg / kg) or a placebo, administered slowly over one hour per study unit. There were no differences in reference ADM levels among the four groups. The median ADM levels were 7.1 pg / mL in the placebo group, 6.8 pg / mL in the first treatment group (0.5 mg / kg), 5.5 pg / mL in the second treatment group (2 mg / kg), and 7.1 pg / mL in the third treatment group (8 mg / mL). The results showed that ADM levels rapidly increased within the first 1.5 hours after administration of NT-H antibody to healthy human subjects, then leveled off and gradually decreased (Figure 6).

[0233] Example 6 - Bio-ADM in patients infected with coronavirus (SARS-CoV-2) Plasma samples from 12 patients diagnosed with coronavirus (SARS-CoV-2) infection were screened for bio-ADM. Bio-ADM levels were measured using an immunoassay method as described by Weber et al. (2017. JALM 2(2): 222-233). DPP3 levels were measured using an immunoassay method (LIA) as described in a recent report (Rehfeld et al. 2019. JALM 3(6): 943-953). The respective bio-ADM and DPP3 concentrations in each sample are summarized in Table 7. [Table 7]

[0234] Bio-ADM concentrations in samples from patients infected with coronavirus (SARS-CoV-2) ranged from 35 to 437 pg / mL, with a median (IQR) of 109 (56-210) pg / mL. The median plasma bio-ADM (mature ADM-NH2) in samples from (healthy) subjects was 24.7 pg / mL, with a minimum value of 11 pg / mL and a 99th percentile of 43 pg / mL (Marino et al. 2014. Critical Care 18:R34). Bio-ADM levels in patients infected with coronavirus (SARS-CoV-2) were significantly elevated compared to healthy controls.

[0235] DPP3 concentrations ranged from 27 to 975 ng / mL, with a median (IQR) of 156.0 (59.5 to 322.3) ng / mL. DPP3 concentrations were significantly elevated compared to healthy controls. Samples were measured from 5,400 normal (healthy) subjects (a single-center prospective population study in Sweden (MPP-RES)), and the median (interquartile range) of plasma DPP3 was 14.5 ng / mL (11.3 ng / mL to 19 ng / mL).

[0236] Example 7 - Changes in lung function in patients with pulmonary dysfunction treated with NT-ADM antibody (AdrenOSS-2) AdrenOSS-2 is a double-blind, placebo-controlled, randomized, multicenter, demonstration and phase II dose-clinical study to investigate the safety, tolerability, and efficacy of an N-terminal ADM antibody named adrecizumab in patients with septic shock and high adrenomedullin levels (Geven et al. BMJ Open 2019;9:e024475). A total of 301 patients with septic shock and bio-ADM concentrations exceeding 70 pg / mL were randomly assigned (2:1:1) to receive a single intravenous infusion lasting approximately one hour with either placebo (152 patients), 2 ng / kg adrecizumab (72 patients), or 4 ng / kg adrecizumab (77 patients). The all-cause mortality rate within 28 (90) days after infusion was 25.8% (34.8%). The mean age was 68.4 years, and 61% were male. Analysis of each methodology showed that 294 patients remained eligible for the trial, and the all-cause mortality rate at day 14 was 18.5%.

[0237] In patients treated with adrecizumab (using a combination of two doses for each group of treatment methods), a tendency toward a lower short-term mortality rate (14 days post-admission) was observed compared to placebo (hazard ratio (HR): 0.701 [0.408~1.21], p=0.200).

[0238] Furthermore, different subgroups were analyzed. The main outcomes were defined as mortality on day 28, changes in the Horowitz index (PaO2 / FiO2) (at 24 / 48 / 72 hours), changes in the SOFA score (at 24 / 48 / 72 hours), or changes in the respiratory SOFA score component (also based on PaO2 / FiO2) (at 24 / 48 / 72 hours). All p-values ​​were two-sided, and a p-value of 0.20 should be considered statistically significant.

[0239] A subgroup (48 patients) of shock patients meeting the criteria of a Horowitz index of less than 170 and standard mechanical ventilation was analyzed. This group mimicked severely ill COVID-19 patients requiring mechanical ventilation in the ICU. Mortality at day 28 tended to be lower in patients treated with adrecizumab compared to placebo (p=0.37) (Figure 7). Changes in the Horowitz index were significantly higher in patients treated with adrecizumab at 48 hours (p=0.09) and 72 hours (p=0.11) (Figures 8B and C), with mean increases of 64.2 and 66.4, respectively, and tended to be higher at 24 hours (0.48) (Figure 8A). The change in SOFA score was significantly lower in patients treated with adrecizumab compared to the placebo group at 24 hours (p=0.032), 48 hours (p=0.012), and 72 hours (p=0.028), respectively (Figures 9A, B, and C).

[0240] A subgroup (80 patients) of shock patients with ALI / ARDS, defined by respiratory diagnosis at admission, was further analyzed. Changes in SOFA scores were significantly lower in patients treated with adrecizumab compared to the placebo group at 24 hours (p=0.005) and 48 hours (p=0.025) (Figure 10A, B), and tended to be lower at 72 hours (p=0.38) (Figure 10C). Furthermore, changes in respiratory SOFA scores in ALI / ARDS patients were significantly lower at 48 hours (p=0.09) and tended to be lower at 24 hours (p=0.26) compared to the placebo group in patients treated with adrecizumab (Figure 11A, B).

[0241] A separate subgroup of shock patients meeting the criteria for standard mechanical ventilation was selected (161 patients). The mortality rate at day 28 was significantly lower in patients treated with adrecizumab compared to the placebo group (p=0.157) (Figure 12). The change in the Horowitz index was significantly higher in patients treated with adrecizumab at 24 hours (p=0.155), 48 hours (p=0.007), and 72 hours (p=0.087), respectively (Figures 13A, B, and C), with a mean increase of 56.2 at 24 hours. Furthermore, the change in SOFA score in patients using standard mechanical ventilation was significantly lower in patients treated with adrecizumab compared to the placebo group at 24 hours (p=0.002) and 48 hours (p=0.109), and tended to be lower at 72 hours (p=0.31) (Figures 14A, B, and C). In particular, the change in respiratory SOFA score in patients using standard ventilators was significantly lower in patients treated with adrecizumab compared to the placebo group at 24 hours (p=0.021), 48 hours (p=0.011), and 72 hours (p=0.066) (Figures 15A, B, and C). These data strongly support the possibility that NT-ADM antibodies can improve endothelial function and vascular integrity in critically ill patients with impaired lung function and suggest their potential applicability to COVID-19 patients.

[0242] Example 8 - Bio-ADM values ​​for prognosis assessment in severely ill COVID-19 patients The purpose of this study was to determine whether bioactive adrenomedullin (bio-ADM) can support risk stratification and clinical management of severely ill COVID-19 patients. 8.1. Study Group and Data Collection

[0243] Following ethical approval (RWTH University Ethics Committee, EK100 / 20), this prospective observational study was conducted at RWTH Aachen University Hospital in Germany from March 13 to April 16, 2020. All patients or their legal representatives provided written consent based on prior notification. All patients who tested positive for SARS-CoV-2 via PCR and were admitted to the ICU were included in this study, except for those under 18 years of age, pregnant patients, and those receiving palliative care. Analysis was performed using real-time reverse transcription PCR (RT-PCR). Patient treatment followed the inventors' standard ICU management, including mechanical ventilation, veno-venous ECMO, and, if necessary, RRT and norepinephrine. Decisions regarding the use of veno-venous ECMO therapy were based on the recently published consensus guidelines for extracorporeal life support systems (ELSO) (Bartlett et al. 2020. ASAIO Journal 66: 472-474). All parameters, including personal statistics, vital signs, laboratory values, blood gas analysis, and organ support, were extracted from the patient data management system (Intellispace Critical Care and Anesthesia (ICCA) system, Philips, Netherlands).

[0244] 8.2. Bio-ADM Measurement Blood samples were collected daily from the day of admission and up to day 7 for analysis of bio-ADM and standard laboratory parameters. Bio-ADM was measured in EDTA plasma using a one-step chemiluminescence sandwich immunoassay (Weber et al. 2017. JALM 2(2): 222-233). Briefly, 100 μL of the sample was cultured at room temperature for 1 hour with agitation in a microtiter plate coated with a monoclonal antibody directed at the central region of bio-ADM, along with 150 μL of detection antibody directed at the C-terminus of bio-ADM. Synthetic human bio-ADM was used as the calibrator. After washing, the chemiluminescence signal was measured in a microtiter plate chemiluminescence reader (Centro LB960, Berthold Technologies, Bad Wildbad, Germany). The detection limit of the assay was 3 pg / mL. In a control group of 200 healthy individuals, the median (99th percentile) bioADM level was 20.7 pg / mL (43 pg / mL) (Marino et al. 2014. Critical Care 18: R34).

[0245] 8.3.Statistical analysis Numerical values ​​were expressed as medians and interquartile ranges (IQR), or as counts and percentages, as needed. Group comparisons of continuous variables were performed using the Kruskal-Wallis test. Categorical data were compared using Pearson's chi-squared test on count data. Biomarker data were logarithmically transformed. Box plots were used to show differences in bioADM within categorical variables. The effect of (logarithmically transformed) bioADM on survival rates in univariate analysis was analyzed using Cox proportional hazards regression. The proportional hazards assumption was tested. Model predictions were evaluated using the chi-squared statistic of the model likelihood ratio. The coincident index (C-index) was obtained as a measure of influence. This is equivalent to the concept of AUC adopted for binary outcomes. Survival curves plotted using the Kaplan-Meier method were used for explanatory purposes. All statistical tests were two-tailed tests, and a two-tailed p-value of 0.05 was considered significant. Statistical analysis was performed using R version 3.4.3 (http: / / www.r-project.org, library rms, Hmisc, ROCR) and Statistical Package for the Social Sciences (SPSS) version 22.0 (SPSS Inc., Chicago, Illinois, USA).

[0246] 8.4.Results This group study successively included 53 COVID-19 patients (40 males [76%], median [IQR]: age 62 years [57–70 years]) after SARS-CoV-2 infection was confirmed and admission to the ICU was required (Table 7). The median length of ICU stay was 16 (7.5–20) days. 32 patients (60%) were discharged from the ICU to a regular ward by day 28, 8 patients (15%) remained in the ICU, and 13 patients (25%) died. Markers of systemic inflammation are shown in Table 7. [Table 8-1] [Table 8-2]

[0247] Variables are shown as median [interquartile range] or numerical value (%). The abbreviations represent the following: ARDS, acute respiratory spurt syndrome; bio-ADM, bioactive adrenomedullin; COPD, chronic obstructive pulmonary disease; CRP, C-reactive protein; ECMO, extracorporeal membrane oxygenation; FiO2, fraction of inspired oxygen; ICU, intensive care unit; IL-6, interleukin-6; pCO2, partial pressure of carbon dioxide; PCT, procalcitonin; PEEP, positive end-expiratory pressure; pO2, partial pressure of oxygen; RRT, renal replacement therapy; spO2, peripheral capillary oxygen saturation; SOFA, sequential organ failure assessment; WBC, white blood cell count.

[0248] A high proportion of the 38 patients (72%) presented with moderate or severe ARDS (25% moderate, 47% severe). Bio-ADM levels increased with the severity of ARDS (p<0.001; bio-ADM levels were 28.3[19.9~28.4], 39.0[29.2~54.5], 48.1[26.9~79.8], and 101.9[67.0~201.1] pg / mL for asymptomatic ARDS, mild ARDS, moderate ARDS, and severe ARDS, respectively) (Figure 16).

[0249] The majority of patients (44) received invasive mechanical ventilation during their ICU stay (Table 7). Bio-ADM levels were significantly elevated in patients on invasive mechanical ventilation compared to patients with spontaneous breathing (68.2 [45.5~106.6] pg / mL vs. 31.8 [18.6~48.4] pg / mL, p=0.006) (Figure 17A). Notably, bio-ADM levels were similarly elevated in patients receiving invasive mechanical ventilation at enrollment (38) compared to patients who required mechanical ventilation during the study (6) (69.8 [44.1~107.3] pg / mL vs. 63.2 [51.0~88.7] pg / mL).

[0250] In patients treated with intravenous ECMO (9 patients), elevated bio-ADM levels were observed compared to patients not receiving ECMO treatment (101.9 [65.0~144.1] pg / mL vs. 53.3 [29.2~91.0] pg / mL, p=0.040) (Figure 17B). In particular, according to the ELSO consensus guidelines, the highest bio-ADM levels were observed in patients who were candidates for ECMO therapy due to the severity of their respiratory failure (7 patients) (Bartlett et al. 2020. ASAIO Journal 66: 472-474), but these patients were not treated with ECMO based on individual patient judgment (262.1 [136.1~274.6] pg / mL, p<0.001) (Figure 17B). Furthermore, bio-ADM levels were significantly correlated with norepinephrine dosage (r=0.47, p<0.001). Patients not receiving norepinephrine had the lowest bio-ADM levels (15 patients, median 37.9 pg / mL), patients receiving low doses of norepinephrine (≤0.1 μg / kg / min, 15 patients) had slightly elevated bio-ADM levels (median 53.8 pg / mL), and patients receiving high doses of norepinephrine (≥0.1 μg / kg / min, 23 patients) had the highest bio-ADM levels (median 105.9 pg / mL, p=0.002).

[0251] Regarding renal function, there was a significant correlation between bio-ADM and serum creatinine (r=0.62, p<0.001). Similarly, patients receiving RRT (27 patients) had significantly higher bio-ADM levels compared to patients not receiving RRT (26 patients) (101.9[67.7~182.9]pg / mL vs. 40.2[27.2~53.5]pg / mL, p<0.001) (Figure 17C).

[0252] Bio-ADM levels were higher in non-survivors (13 people) than in survivors (40 people) (107.6 [51.0~262.1] pg / mL vs. 53.3 [29.2~91.0] pg / mL, p=0.010). In particular, bio-ADM predicted mortality at day 28 (C index 0.72, 95% confidence interval [CI] 0.56~0.87, p<0.001) (Figure 18A).

[0253] The inventors then further clarified the values ​​by continuously measuring bio-ADM and predicted the mortality rate at day 28. Based on previous studies (Mebazaa et al. 2018. Critical Care 22: 354), a bio-ADM cutoff value of 70 pg / mL was used, and patients were grouped accordingly. Patients with bio-ADM levels above 70 pg / mL at enrollment (high) and who maintained that level (high → high) showed the worst outcomes, while patients who improved within 48 hours (high → low) showed favorable outcomes. Similarly, patients showing an increase in bio-ADM within 48 hours (low → high) showed unfavorable results (Figure 18B).

[0254] In conclusion, bio-ADM plasma levels correlate with disease severity, the need for extracorporeal organ support, and outcomes, highlighting the favorable prognosis of bio-ADM in early risk stratification and management of COVID-19 patients. Furthermore, the data clearly highlight the role of endothelial dysfunction in the pathophysiology of COVID-19 and pave the way for future randomized trials prospectively evaluating bio-ADM as a novel objective tool for risk stratification and monitoring of patients with COVID-19. [Brief explanation of the drawing]

[0255] [Figure 1a] Figure 1a is an explanatory diagram of antibody form:Fv isomers and scFv variants. [Figure 1b] Figure 1b is an explanatory diagram of antibody types: heterogeneous fusion antibodies and bifunctional antibodies. [Figure 1c] Figure 1c is an explanatory diagram of antibody types: bivalent antibodies and bispecific antibodies. [Figure 2ab] Figure 2 shows the following: a: Dose-response curve of human ADM. Maximum cAMP stimulation was adjusted to 100% activity. b: Dose / inhibition rate curve of human ADM22-52 (ADM receptor antagonist) in the presence of 5.63 nM hADM. [Figure 2cd]c: Shows the dose / inhibition rate curve for CT-H in the presence of 5.63 nM hADM. d: Shows the dose / inhibition rate curve for MR-H in the presence of 5.63 nM hADM. [Figure 2ef] e: Shows the dose / inhibition rate curve of NT-H in the presence of 5.63 nM hADM. f: Shows the dose-response curve of mouse ADM. Maximum cAMP stimulation was adjusted to 100% activity. [Figure 2gh] g: Dose / inhibition rate curves for human ADM 22-52 (ADM receptor antagonists) in the presence of 0.67 nM mADM. h: Dose / inhibition rate curves for CT-M in the presence of 0.67 nM mADM. [Figure 2ij] i: Shows the dose / inhibition rate curve for MR-M in the presence of 0.67 nM mADM. j: Shows the dose / inhibition rate curve for NT-M in the presence of 0.67 nM mADM. [Figure 2kl] k: Shows the inhibition rate of ADM by F(ab)2NT-M and Fab NT-M. l: Shows the inhibition rate of ADM by F(ab)2NT-M and Fab NT-M. [Figure 3] Figure 3 shows a typical hADM dose / signal curve, as well as the hADM dose / signal curve in the presence of 100 μg / mL NT-H antibody. [Figure 4] Figure 4 shows the stability of hADM in human plasma (citrate) with and without NT-H antibody. [Figure 5] Figure 5 shows the alignment of Fabs having homologous human framework sequences. [Figure 6] Figure 6 shows the ADM concentrations in healthy human subjects up to 60 days after administration of NT-H at various doses. [Figure 7] Figure 7 shows the 28-day mortality rate for patients with a Horowitz index of less than 170 and using a standard ventilator (n=48). [Figure 8ab] Figure 8 shows (A) 24 hours, (B) 48 hours, and , for patients (n=48) with a Horowitz index of less than 170 and using a standard ventilator. [Figure 8c](C) Shows the change in the Horowitz index after 72 hours. [Figure 9ab] Figure 9 shows (A) 24 hours, (B) 48 hours, and , for patients (n=48) with a Horowitz index of less than 170 and using a standard ventilator. [Figure 9c] (C) Shows the change in SOFA score after 72 hours. [Figure 10ab] Figure 10 shows (A) 24 hours, (B) 48 hours, and , ALI / ARDS patients (n=80) [Figure 10c] (C) Shows the change in SOFA score after 72 hours. [Figure 11] Figure 11 shows the changes in respiratory SOFA scores at (A) 24 hours and (B) 48 hours, respectively, in ALI / ARDS patients (n=80). [Figure 12] Figure 12 shows the 28-day mortality rate for patients using standard ventilators (n=161). [Figure 13ab] Figure 13 shows (A) 24 hours, (B) 48 hours, and for patients using a standard ventilator (n=161). [Figure 13c] (C) Shows the change in the Horowitz index after 72 hours. [Figure 14ab] Figure 14 shows (A) 24 hours, (B) 48 hours, and for patients using a standard ventilator (n=161). [Figure 14c] (C) Shows the change in SOFA score after 72 hours. [Figure 15ab] Figure 15 shows (A) 24 hours, (B) 48 hours, and for patients using a standard ventilator (n=161). [Figure 15c] (C) Shows the change in respiratory SOFA score after 72 hours for each individual. [Figure 16] Figure 16 shows a box plot of bio-ADM due to ARDS in 53 COVID-19 patients (p<0.001). The horizontal line in the figure represents 70 pg / mL. [Figure 17a]Figure 17 shows box plots of bio-ADM levels in 53 COVID-19 patients treated with invasive mechanical ventilation (A, p=0.006), ECMO (B, p<0.001), and RRT (C, p<0.001). The horizontal line in the figure represents 70 pg / mL. Patients who meet the criteria for ECMO treatment but do not receive ECMO treatment are referred to as "indicated." [Figure 17bc] Figure 17 shows box plots of bio-ADM levels in 53 COVID-19 patients treated with invasive mechanical ventilation (A, p=0.006), ECMO (B, p<0.001), and RRT (C, p<0.001). The horizontal line in the figure represents 70 pg / mL. Patients who meet the criteria for ECMO treatment but do not receive ECMO treatment are referred to as "indicated." [Figure 18ab] Figure 18 shows Kaplan-Meier plots of 28-day mortality for bio-ADM. (A) Curves are plotted by the quartile of bio-ADM (for continuous bio-ADM, the standardized HR is 3.5 (95% CI 1.6–7.5), the c-index is 0.72 (95% CI 0.56–0.87), and p<0.001). (B) Curves are plotted to show the potential value of continuous bio-ADM measurements by whether the value is greater than or less than 70 pg / mL at enrollment and 48 hours (right side, p=ns). Outcomes are worst for patients with bio-ADM greater than and remaining above 70 pg / mL at enrollment (high → high), but good for patients who improved within 48 hours (high → low). Similarly, patients showing an increase in bio-ADM (low → high) have unfavorable outcomes. Patients for whom bio-ADM data is missing at 48 hours will remain in their initial category.

[0256] array Sequence ID 1: GYTFSRYW Sequence ID 2: ILPGSGST Sequence ID 3: TEGYEYDGFDY Sequence ID 4: QSIVYSNGNTY Array "RVS" (not part of the array list): RVS Sequence ID 5: FQGSHIPYT Sequence number 6 (AM-VH-C): [ka] Sequence ID 7 (AM-VH1): [ka] Sequence ID 8 (AM-VH2-E40): [ka] Sequence ID 9 (AM-VH3-T26-E55): [ka] Sequence ID 10 (AM-VH4-T26-E40-E55): [ka] Sequence ID 11(AM-VL-C): [ka] Sequence ID 12 (AM-VL1): [ka] Sequence ID 13 (AM-VL2-E40): [ka] Sequence ID 14 (Human ADM 1-21): YRQSMNNFQGLRSFGCRFGTC Sequence ID 15 (Human ADM 21-32): CTVQKLAHQIYQ Sequence ID 16 (Human ADM C-42~52): CAPRSKISPQGY-CONH2 Sequence ID 17 (Mouse ADM 1-19): YRQSMNQGSRSNGCRFGTC Sequence ID 18 (Mouse ADM 19-31): CTFQKLAHQIYQ Sequence ID 19 (Mouse ADM C40~50): CAPRNKISPQGY-CONH2 Sequence ID No. 20 (Mature Human Adrenomedullin (Mature ADM); Amidated ADM; Bio-ADM): Amino acids 1-52 or 95-146 of pro-ADM: [ka] SEQ ID NO: 21 (Adrenomedullin 1-52-Gly (ADM 1-52-Gly): Amino acids 95-147 of prepro-ADM) [ka] Sequence ID 22 (Mouse ADM 1-50): [ka] Sequence ID 23 (Human ADM 1-42): [ka] Sequence ID No. 24 (Amino acids 43-52 of human ADM): PRSKISPQGY-NH2 Sequence ID No. 25 (Amino acids 1-14 of human ADM): YRQSMNNFQGLRSF Sequence ID 26 (Amino acids 1-10 of human ADM): YRQSMNNFQG Sequence ID 27 (Amino acids 1-6 of human ADM): YRQSMN Sequence ID No. 28 (Amino acids 1-32 of human ADM): [ka] Sequence ID 29 (Amino acids 1-40 of mouse ADM): [ka] Sequence ID 30 (Amino acids 1-31 of mouse ADM): [ka] Sequence ID 31 (pro-ADM; 164 amino acids (22-185 of prepro-ADM)): [ka] Sequence ID 32 (Proadrenomedullin N-20 terminal peptide, PAMP: amino acids 22-41 of prepro-ADM): ARLDVASEF RKKWNKWALS R SEQ ID NO: 33 (Central region proadrenomedullin, MR-proADM: amino acids 45-92 of prepro-ADM): [ka] SEQ ID NO: 34 (C-terminal proadrenomedullin, CT-proADM: amino acids 148-185 of prepro-ADM): [ka] Sequence ID 35 (heavy chain, HAM8101): [ka] Sequence ID 36 (light chain, HAM8101): [ka] Sequence ID 37-IGHV1-69*11: [ka] Sequence ID 38-HB3: [ka] Some aspects of the present invention are described below. 1. Anti-adrenomedullin (ADM) antibodies, anti-ADM antibody fragments, or anti-ADM non-Ig scaffolds for use in the treatment or intervention of patients infected with coronavirus. 2. An anti-adrenomedullin (ADM) antibody or anti-ADM antibody fragment or anti-ADM non-Ig scaffold for use in the treatment or intervention of a patient infected with the coronavirus described in item 1, wherein the coronavirus is selected from the group including Sars-CoV-1, Sars-CoV-2, and MERS-CoV, and in particular Sars-CoV-2. 3. Anti-adrenomedullin (ADM) antibody or anti-ADM antibody fragment or anti-ADM non-Ig scaffold for use in the treatment or intervention of a patient infected with the coronavirus described in item 1 or 2, wherein the patient has a level of proadrenomedullin or a fragment thereof in the subject body fluid sample that exceeds a predetermined threshold for proadrenomedullin or is higher than the level of proadrenomedullin measured previously. 4. An anti-adrenomedullin (ADM) antibody, an anti-ADM antibody fragment, or an anti-ADM non-Ig scaffold for use in the treatment or intervention of a patient infected with the coronavirus described in any of items 1 to 3, wherein the patient has a D-dimer level of 0.5 μg / ml or higher, preferably 1.0 μg / ml or higher. 5. An anti-adrenomedullin (ADM) antibody, anti-ADM antibody fragment, or anti-ADM non-Ig scaffold for use in the treatment or intervention of a patient infected with the coronavirus described in any of items 1 to 4, wherein the anti-ADM antibody or anti-ADM fragment or anti-ADM non-Ig scaffold binds to the N-terminal portion (amino acids 1-21) of ADM-Gly and / or ADM-NH2: YRQSMNNFQGLRSFGCRFGTC (SEQ ID NO: 14). 6. An anti-adrenomedullin (ADM) antibody or anti-ADM antibody fragment or anti-ADM non-Ig scaffold for use in the treatment or intervention of a patient infected with any of the coronaviruses described in items 1 to 5, wherein the antibody is a monoclonal antibody or a monoclonal antibody fragment. 7. The heavy chain complementarity-determining region (CDR) is the following sequence: CDR1: Sequence ID 1 GYTFSRYW CDR2: Sequence ID 2 ILPGSGST CDR3: Sequence ID 3 TEGYEYDGFDY Includes, And the complementarity-determining region (CDR) of the light chain is the following sequence: CDR1: Sequence ID 4 QSIVYSNGNTY CDR2: RVS CDR3: Sequence ID 5 FQGSHIPYT including, Anti-adrenomedullin (ADM) antibodies, anti-ADM antibody fragments, or anti-ADM non-Ig scaffolds for use in the treatment or intervention of patients infected with the coronavirus as described in item 6. 8. The antibody or fragment, as a VH region, Sequence number 6 (AM-VH-C) QVQLQQSGAELMKPGASVKISKATGYTFSRYWIEWVKQRPGHGLEWIGEILPGSGSTNYNEKFKGKATITADTSSNTAYMQLSSLTSEDSAVYYCTEGYEYDGFDYWG QGTTLTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPK Sequence ID 7 (AM-VH1) QVQLVQSGAEVKKPGSSVKVSCKASGYTFSRYWISWVRQAPGQGLEWMGRILPGSGSTNYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCTEGYEYDGFDYWG QGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPK Sequence ID 8 (AM-VH2-E40) QVQLVQSGAEVKKPGSSVKVSCKASGYTFSRYWIEWVRQAPGQGLEWMGRILPGSGSTNYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCTEGYEYDGFDYWG QGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPK Sequence ID 9 (AM-VH3-T26-E55) QVQLVQSGAEVKKPGSSVKVSCKATGYTFSRYWISWVRQAPGQGLEWMGEILPGSGSTNYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCTEGYEYDGFDYWG QGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPK Sequence ID 10 (AM-VH4-T26-E40-E55) QVQLVQSGAEVKKPGSSVKVSCKATGYTFSRYWIEWVRQAPGQGLEWMGEILPGSGSTNYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCTEGYEYDGFDYWG QGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPK Alternatively, the sequence may be selected from a group that includes sequences that are >80% identical to the sequences described above. And the following sequence as the VL region: Sequence ID 11 (AM-VL-C) DVLLSQTPLSLPVSLGDQATISCRSSQSIVYSNGNTYLEWYLQKPGQSPKLLIYRVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDLGVYYCFQGSHIPYTFGGGTKL EIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC Sequence ID 12 (AM-VL1) DVVMTQSPLSLPVTLGQPASISCRSSQSIVYSNGNTYLNWFQQRPGQSPRRLIYRVSNRDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFQGSHIPYTFGQGTKL EIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC Sequence ID 13 (AM-VL2-E40) DVVMTQSPLSLPVTLGQPASISCRSSQSIVYSNGNTYLEWFQQRPGQSPRRLIYRVSNRDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFQGSHIPYTFGQGTKL EIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC Alternatively, the sequence may be selected from a group that includes sequences that are >80% identical to the sequences described above. Anti-adrenomedullin (ADM) antibodies, anti-ADM antibody fragments, or anti-ADM non-Ig scaffolds for use in the treatment or intervention of patients infected with the coronavirus as described in item 7. 9. The antibody or fragment, as a heavy chain, Sequence ID 35 QVQLVQSGAEVKKPGSSVKVSCKASGYTFSRYWIEWVRQAPGQGLEWIGEILPGSGSTNYNQKFQGRVTITADTSTSTAYMELSSLRSEDTAVYYCTEGYEYDGFDYWGQGT TVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKT HTCPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKT ISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK or including a sequence that is >95% identical to the said sequence, And the following sequence as the light chain: Sequence ID 36 DVVLTQSPLSLPVTLGQPASISCRSSQSIVYSNGNTYLEWYLQRPGQSPRLLIYRVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFQGSHIPYTFGGGTKL EIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC or including a sequence that is >95% identical to the said sequence, Anti-adrenomedullin (ADM) antibodies or anti-ADM antibody fragments or anti-ADM non-Ig scaffolds for use in the treatment or intervention of patients infected with the coronavirus as described in item 7 or 8. 10. Anti-adrenomedullin (ADM) antibodies or anti-ADM antibody fragments or anti-ADM non-Ig scaffolds for use in the treatment or intervention of patients with impaired pulmonary function and / or acute respiratory distress syndrome (ARDS). 11. Anti-adrenomedullin (ADM) antibody or anti-ADM antibody fragment or anti-ADM non-Ig scaffold for use in the treatment or intervention of patients having impaired pulmonary function and / or acute respiratory distress syndrome (ARDS) as described in item 10, in which the patient has a Horowitz index of 300 or less, particularly 200 or less, particularly 100 or less, and / or the patient requires mechanical ventilation. 12. An anti-adrenomedullin (ADM) antibody or anti-ADM antibody fragment or anti-ADM non-Ig scaffold for use in the treatment or intervention of a patient having impaired pulmonary function and / or acute respiratory distress syndrome (ARDS) as described in item 10 or 11, wherein the patient has a level of proadrenomedullin or a fragment thereof in the subject fluid sample that exceeds a predetermined threshold for proadrenomedullin or is higher than a previously measured level of proadrenomedullin. 13. The heavy chain complementarity-determining region (CDR) is the following sequence: CDR1: Sequence ID 1 GYTFSRYW CDR2: Sequence ID 2 ILPGSGST CDR3: Sequence ID 3 TEGYEYDGFDY Includes, And the complementarity-determining region (CDR) of the light chain is the following sequence CDR1: Sequence ID 4 QSIVYSNGNTY CDR2: RVS CDR3: Sequence ID 5 FQGSHIPYT including, An anti-adrenomedullin (ADM) antibody, anti-ADM antibody fragment, or anti-ADM non-Ig scaffold for use in the treatment or intervention of patients with impaired pulmonary function and / or acute respiratory distress syndrome (ARDS) as described in any of items 10-12. 14. The antibody or fragment, as a VH region, Sequence number 6 (AM-VH-C) QVQLQQSGAELMKPGASVKISKATGYTFSRYWIEWVKQRPGHGLEWIGEILPGSGSTNYNEKFKGKATITADTSSNTAYMQLSSLTSEDSAVYYCTEGYEYDGFDYWG QGTTLTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPK Sequence ID 7 (AM-VH1) QVQLVQSGAEVKKPGSSVKVSCKASGYTFSRYWISWVRQAPGQGLEWMGRILPGSGSTNYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCTEGYEYDGFDYWG QGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPK Sequence ID 8 (AM-VH2-E40) QVQLVQSGAEVKKPGSSVKVSCKASGYTFSRYWIEWVRQAPGQGLEWMGRILPGSGSTNYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCTEGYEYDGFDYWG QGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPK Sequence ID 9 (AM-VH3-T26-E55) QVQLVQSGAEVKKPGSSVKVSCKATGYTFSRYWISWVRQAPGQGLEWMGEILPGSGSTNYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCTEGYEYDGFDYWG QGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPK Sequence ID 10 (AM-VH4-T26-E40-E55) QVQLVQSGAEVKKPGSSVKVSCKATGYTFSRYWIEWVRQAPGQGLEWMGEILPGSGSTNYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCTEGYEYDGFDYWG QGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPK Alternatively, the sequence may be selected from a group that includes sequences that are >80% identical to the sequences described above. And as the VL region, the following sequence Sequence ID 11 (AM-VL-C) DVLLSQTPLSLPVSLGDQATISCRSSQSIVYSNGNTYLEWYLQKPGQSPKLLIYRVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDLGVYYCFQGSHIPYTFGGGTKL EIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC Sequence ID 12 (AM-VL1) DVVMTQSPLSLPVTLGQPASISCRSSQSIVYSNGNTYLNWFQQRPGQSPRRLIYRVSNRDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFQGSHIPYTFGQGTKL EIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC Sequence ID 13 (AM-VL2-E40) DVVMTQSPLSLPVTLGQPASISCRSSQSIVYSNGNTYLEWFQQRPGQSPRRLIYRVSNRDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFQGSHIPYTFGQGTKL EIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC Alternatively, the sequence may be selected from a group that includes sequences that are >80% identical to the sequences described above. Anti-adrenomedullin (ADM) antibodies, anti-ADM antibody fragments, or anti-ADM non-Ig scaffolds for use in the treatment or intervention of patients with impaired pulmonary function and / or acute respiratory distress syndrome (ARDS) as described in item 13. 15. The antibody or fragment has the following sequence as its heavy chain: Sequence ID 35 QVQLVQSGAEVKKPGSSVKVSCKASGYTFSRYWIEWVRQAPGQGLEWIGEILPGSGSTNYNQKFQGRVTITADTSTSTAYMELSSLRSEDTAVYYCTEGYEYDGFDYWGQGT TVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKT HTCPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKT ISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK or including a sequence that is >95% identical to the said sequence, And as a light chain, Sequence ID 36 DVVLTQSPLSLPVTLGQPASISCRSSQSIVYSNGNTYLEWYLQRPGQSPRLLIYRVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFQGSHIPYTFGGGTKL EIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC or including a sequence that is >95% identical to the said sequence, Anti-adrenomedullin (ADM) antibodies, anti-ADM antibody fragments, or anti-ADM non-Ig scaffolds for use in the treatment or intervention of patients with impaired pulmonary function and / or acute respiratory distress syndrome (ARDS) as described in items 13-14.

Claims

1. A composition comprising an anti-adrenomedullin (ADM) antibody or an anti-ADM antibody fragment for use in the treatment of patients with pulmonary dysfunction and / or acute respiratory distress syndrome (ARDS), wherein the anti-ADM antibody or anti-ADM antibody fragment is directed to and bound to the N-terminal portion (amino acids 1-21) of ADM: YRQSMNNNFQGLRSFGCRFGTC (SEQ ID NO: 14).

2. A composition comprising an anti-adrenomedullin (ADM) antibody or an anti-ADM antibody fragment for use in the treatment of a patient having pulmonary dysfunction and / or acute respiratory distress syndrome (ARDS) as described in claim 1, wherein the patient has a Horowitz index of 300 or less and / or the patient requires a ventilator.

3. A composition comprising an anti-adrenomedullin (ADM) antibody or an anti-ADM antibody fragment for use in the treatment of a patient having pulmonary dysfunction and / or acute respiratory distress syndrome (ARDS) as described in claim 2, wherein the patient has a Horowitz index of 200 or less.

4. A composition comprising an anti-adrenomedullin (ADM) antibody or an anti-ADM antibody fragment for use in the treatment of a patient having pulmonary dysfunction and / or acute respiratory distress syndrome (ARDS) as described in claim 3, wherein the patient has a Horowitz index of 100 or less.

5. A composition for use in treating a patient having pulmonary dysfunction and / or acute respiratory distress syndrome (ARDS) according to any one of claims 1 to 4, wherein the patient has a level of proadrenomedullin or a fragment thereof in a patient's body fluid sample that exceeds a predetermined threshold for proadrenomedullin or is higher than the level of proadrenomedullin in a previously measured body fluid sample of the patient, wherein the body fluid sample is blood, serum, or plasma, and the proadrenomedullin fragment is PAMP (SEQ ID NO: 32), MR-proADM (SEQ ID NO: 33), ADM-NH 2 A composition selected from the group consisting of (SEQ ID NO: 20), ADM-Gly (SEQ ID NO: 21), and CT-proADM (SEQ ID NO: 34).

6. The heavy chain complementarity-determining region (CDR) is the following sequence: CDR1: Sequence ID 1 GYTFSRYW CDR2: Sequence ID 2 ILPGSGST CDR3: Sequence ID 3 TEGYEYDGFDY Includes, And the complementarity-determining region (CDR) of the light chain is the following sequence CDR1: Sequence ID 4 QSIVYSNGNTY CDR2: RVS CDR3: Sequence ID 5 FQGSHIPYT including, A composition comprising an anti-adrenomedullin (ADM) antibody or an anti-ADM antibody fragment for use in the treatment of a patient having pulmonary dysfunction and / or acute respiratory distress syndrome (ARDS) as described in any one of claims 1 to 5.

7. The antibody or fragment has the following sequence as its heavy chain: Sequence ID 35 QVQLVQSGAEVKKPGSSVKVSCKASGYTFSRYWIEWVRQAPGQGLEWIGEILPGSGSTNYNQKFQGRVTITADTSTSTAYMELSSLRSEDTAVYYCTEGYEYDGFDYWGQGT TVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKT HTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKT ISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK or including a sequence that is >95% identical to the said sequence, And the following sequence as the light chain: Sequence ID 36 DVVLTQSPLSLPVTLGQPASISCRSSQSIVYSNGNTYLEWYLQRPGQSPRLLIYRVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFQGSHIPYTFGGGTKL EIKRTVAAPSVFIFPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC or including a sequence that is >95% identical to the said sequence, A composition comprising an anti-adrenomedullin (ADM) antibody or an anti-ADM antibody fragment for use in the treatment of a patient having pulmonary dysfunction and / or acute respiratory distress syndrome (ARDS) as described in claim 6.

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