NZ624869B2 - Anti-adrenomedullin (adm) antibody or anti-adm antibody fragment or anti-adm non-ig scaffold for reducing the risk of mortality in a patient having a chronic or acute disease or acute condition - Google Patents

Abstract

Discloses the use of an anti-adrenomedullin (ADM) antibody or an anti-adrenomedullin antibody fragment or an anti-ADM non Ig scaffold binding to adrenomedullins, in the manufacture of a medicament for therapy or prevention of SIRS, meningitis, sepsis or shock in a patient for the reduction of the mortality risk for said patient wherein said antibody fragment or non Ig scaffold binds to the N-terminal part, aa 1-21, of adrenomedullin: YRQSMNNFQGLRSFGCRFGTC (SEQ ID NO: 23). rtality risk for said patient wherein said antibody fragment or non Ig scaffold binds to the N-terminal part, aa 1-21, of adrenomedullin: YRQSMNNFQGLRSFGCRFGTC (SEQ ID NO: 23).

Description

/072929 Anti-Adrenomedullin (ADM) antibody or anti-ADM dy fragment or anti-ADM non-1g scaffold for reducing the risk of mortality in a patient having a chronic or acute disease or acute condition Field of the invention Subject matter of the present invention is an drenomedullin (ADM) antibody or an anti- adrenomedullin antibody fragment or an anti~ADM non—lg ld for use in therapy of a severe chronical or acute disease or acute condition of a patient for the reduction of the mortality risk for said patient. ound The peptide adrenomedullin (ADM) was described for the first time in 1993 (Kitamura, K., et at, "Adrenomedullin: A Novel Hypotensive Peptide Isolated From Human Pheochrornocytoma", Biochemical and Biophysical Research Communications, Vol. 192 (2), pp. 553660 (1993)) as a novel hypotensive peptide comprising 52 amino acids, which had been isolated from a human pheochromocytome; SEQ ID No.: 21. In the same year, cDNA coding for a precursor peptide comprising 185 amino acids and the complete amino acid ce of this precursor peptide were also described. The sor peptide, which comprises, inter alia, a signal sequence of 21 amino acids at the N—terminus, is referred to as "preproadrenomedullin“ (pre—proADM). in the t description, all amino acid positions specified usually relate to the pre—proADM which comprises the 185 amino acids. The peptide adrenomedullin (ADM) is a peptide which comprises 52 amino acids (SEQ ID NO: 21) and which comprises the amino acids 95 to 146 of pre—proADM, from which it is formed by proteolytic cleavage. To date, substantially only a few fragments of the e fragments formed in the cleavage of the premproADM have been more exactly investigated, in particular the physiologically active peptides adrenornedullin (ADM) and “PAMP", a peptide sing 20 amino acids (22-41) which follows the 21 amino acids of the signal e in pre—proADM. The discovery and characterization of ADM in 1993 triggered intensive research activity, the results of which have been summarized in various review articles, in the context of the present description, reference being made in particular to the es to be found in an issue of "Peptides" devoted to ADM in particular (Editorial, Takahashi, K., ”Adrenomedullin: from a pheochrornocytoma to the eyes", Peptides, Vol. 22, p. 1691 (2001)) and (Etc, T., "A review of the biological properties and al implications of adrenomeduilin and proadrenomedullin N—terminal 20 peptide (PAMP), hypotensive and 2012/072929 vasodilating peptides", es, Vol. 22, pp. 1693—1711 (2001)). A further review is (Hinson, at at, "Adrenomedullin, a Multifunctional Regulatory Peptide", Endocrine Reviews, Vol. 21(2), pp. 138-167 (2000)). In the scientific investigations to date, it has been found, inter alia, that ADM may be regarded as a polyfunctional regulatory peptide. It is ed into the circulation in an inactive form extended by glycine (Kitamura, K., at a], "The intermediate form of glycine-extended medullin is the major circulating molecular form in human plasma", Biochem. Biophys. Res. Commun, Vol. , pp. 551—555 (1998). Abstract Only). There is also a binding protein (Pio, R., at at, "Complement Factor H is 21 Serum- binding Protein for adrenomedullin, and the Resulting Complex Modulates the Bioactivities of Both Partners", The Journal of Biological Chemistry, Vol. 276(15), pp. 12292-12300 (2001)) which is specific for ADM and probably likewise modulates the effect of ADM.

Those physiological effects of ADM as well as of PAMP which are of y importance in the igations to date were the s influencing blood re.

Hence, ADM is an effective vasodilator, and thus it is possible to ate the hypotensive effect with the ular peptide segments in the C-terminal part of ADM. It has furthermore been found that the above—mentioned further physiologically active e PAMP formed from pre—proADM likewise exhibits a nsive effect, even if it appears to have an action mechanism ing from that ofADM (of. in addition to the abovementioned review articles (Eto, T., "A review of the biological properties and clinical ations of adrenornedullin and proadrenornedullin N—terminal 20 peptide (PAMP), hypotensive and vasodilating peptides", es, Vol. 22, pp. 1693—1711 (2001)) and (Hinson, et at, 1‘Adrenomedullin, a Multifunctional Regulatory Peptide", Endocrine Reviews, Vol. 21(2), pp. 138—167 (2000)) also (Kuwasako, K., at all, "Purification and characterization of PAMP-l2 (PAMP~20) in porcine adrenal medulla as a major endogenous biologically active peptide“, FEBS Lett, Vol. 414(1), pp. 105—110 (1997). Abstract only), (Kuwasaki, K., at 52]., "Increased plasma proadrenomedullin N—terrninal 20 peptide in patients with essential hypertension", Ann. Clin.

Biochem., Vol. 36 (Pt. 5), pp. 622—628 (1999). Abstract only) or (Tsuruda, T., et at, "Secretion of proadrenomedullin N—terminal 20 peptide from cultured neonatal rat cardiac cells", Life Sci, Vol. 69(2), pp. 239—245 (2001). Abstract only) and EP-A2 0 622 458). It has furthermore been found that the concentrations of ADM which can be measured in the circulation and other biological liquids are, in a number of pathological states, significantly above the concentrations to be found in healthy control persons. Thus, the ADM level in patients with congestive heart e, myocardial infarction, kidney diseases, hypertensive disorders, Diabetes mellitus, in the acute phase of shock and in sepsis and septic shock are significantly increased, although to different extents. The PAMP concentrations are also increased in some of said pathological states, but the plasma levels are lower relative to ADM ((Eto, T., "A review of the biological properties and clinical implications of adrenomedullin and proadrenomedullin N-terminal 20 peptide , hypotensive and vasodilating peptides”, Peptides, Vol. 22, pp. 1693—1711 (2001)); page 1702). It is filrthermore known that unusually high concentrations of ADM are to be observed in , and the highest concentrations in septic shock (cf. (Etc, T., "A review of the biological properties and clinical implications of adrenomedullin and proadrenomedullin inal 20 peptide (PAMP), hypotensive and vasodilating peptides", Peptides, Vol. 22, pp. 1693—1711 (2001)) and (Hirata, at £11., "Increased Circulating medullin, a Novel Vasodilatory Peptide, in Sepsis", Journal of Clinical Endocrinology and Metabolism, Vol. 81(4), pp. 1449—1453 ), (Ehlenz, K., at at, "High levels of circulating adrenomedullin in severe illness: Correlation with C-reactive protein and evidence against the adrenal medulla as site of origin", Exp Clin Endocrinol Diabetes, Vol. 105, pp. l56-162 (1997)), (Tomoda, Y., at 611., "Regulation of adrenomedullin secretion from cultured cells", Peptides, Vol. 22, pp. 1783—1794 (2001)), (Ueda, 8., er al, "Increased Plasma Levels of Adrenomedullin in Patients with Systemic Inflammatory Response Syndrome", Am. J. Respir. Crit. Care Med, Vol- 160, pp. 132-136 (1999)) and (Wang, P., ”Adrenomedullin and vascular responses in ", Peptides, Vol.22, pp. 1835—1840(2001))).

Known in the art is further a method for fying adrenomedullin immunoreactivity in biological s for diagnostic purposes and, in particular within the scope of sepsis sis, cardiac diagnosis and cancer diagnosis. According to the invention, the midregional partial peptide of the proadrenomedullin, which contains amino acids (45-92) of the entire preproadrenomedullin, is measured, in particular, with an immunoassay which works with at least one labeled dy that specifically recognizes a sequence of the mid- proADM (W02004/090546).

WO—Al 2004/097423 describes the use of an antibody against adrenomedullin for diagnosis, prognosis, and treatment of vascular disorders. Treatment of diseases by ng the ADM receptor are also described in the art, (ag. WO—Al 2006/027147, ) said diseases may be sepsis, septic shock, cardiovascular diseases, infections, dermatological diseases, endocrinological diseases, lic es, gastroenterological diseases, cancer, inflammation, hematological diseases, atory diseases, muscle skeleton diseases, neurological diseases, urological diseases.

It is reported for the early phase of sepsis that ADM improves heart fimction and the blood supply in liver, spleen, kidney and small intestine. Anti—ADM-ncutralizing antibodies neutralize the before mentioned effects during the early phase of sepsis (Wang, P., ”Adrenomedullin and cardiovascular responses in sepsis", es, Vol. 22, pp. 1835-1840 (2001).

In the later phase of sepsis, the hypodynainical phase of sepsis, ADM constitutes a risk factor that is strongly associated with the mortality of patients in septic shock. (Schiitz er al., “Circulating sor levels of endothelin-l and adrenomedullin, two endothelium—derived, counteracting substances, in sepsis”, Endothelium, 14:345—351, (2007)). s for the diagnosis and treatment of critically ill patients, rag. in the very late phases of , and the use of endothelin and endothelin agonists with nstrictor activity for the preparation of medicaments for the treatment of critically ill patients have beendescribed in WO-Al 20072062676. It is further described in WO—Al 20077062676 to use, in place of endothelin and/or endothelin agonists, or in ation therewith, adrenomedullin antagonists, zie. molecules which prevent or attenuate the vasodilating action of adrenomedullin, ag. by ng its relevant receptors, or substances preventing the g of adrenomedullin to its receptor (ag. specific binders as cg. antibodies binding to adrenomedullin and blocking its receptor bindings sites; “immunological neutralization“). Such use, or combined use, including a subsequent or preceding separate use, has been described in certain cases to be desirable for example to improve the therapeutic success, or to avoid undesirable physiological stress or side effects. Thus, it is reported that neutralizing anti—ADM antibodies may be used for the treatment of sepsis in the late stage of sepsis.

Administration of ADM in combination with ADM-binding—Proteiml is described for treatment of sepsis and septic shock in the art. It is assumed that treatment of septic s with ADM and ADM—binding-Protein-l prevents transition to the late phase of sepsis. It has to be noted that in a living organism ADM binding n (complement factor H) is present in the circulation of said sm in high concentrations (l’io e: at: fication, characterization, and physiological actions of factor H as an Adrenomedullin g Protein present in Human Plasma; Microscopy Res. and Technique, 55:23‘27 (2002) and Martinez et (followed by 5A) Thus, in one ment the present invention provides for the use of an anti-adrenomedullin (ADM) dy or an anti-adrenomedullin antibody fragment or an anti-ADM non Ig ld binding to adrenomedullins, in the cture of a medicament for therapy or prevention of SIRS, itis, sepsis or shock in a patient for the reduction of the mortality risk for said patient n said antibody fragment or non Ig scaffold binds to the N-terminal part, aa 1- 21, of adrenomedullin: In another embodiment the present invention provides for the use of a vasopressor in the manufacture of a medicament for therapy or prevention of SIRS, meningitis, sepsis or shock in a patient for the reduction of the mortality risk for said patient, wherein said therapy or prevention comprises administration of an anti-adrenomedullin (ADM) antibody or an renomedullin antibody fragment or an anti-ADM non Ig scaffold binding to adrenomedullins in combination with said vasopressor, wherein said antibody fragment or non Ig scaffold binds to the N-terminal part, aa 1-21, of adrenomedullin: In a still further embodiment the present invention provides for the use of intravenous fluids in the cture of a medicament for therapy or prevention of SIRS, meningitis, sepsis or shock in a patient for the reduction of the mortality risk for said patient, wherein said therapy or prevention comprises administration of an anti-adrenomedullin (ADM) antibody or an anti- 5A (followed by page 6) adrenomedullin dy fragment or an anti-ADM non Ig scaffold binding to adrenomedullins in combination with said intravenous fluids, wherein said antibody fragment or non Ig scaffold binds to the N-terminal part, aa 1-21, of adrenomedullin: binding to ADM may not be ADM—binding—Protein—l (complement factor H). ADM-binding— Protein—l is not meant by any of the before mentioned anti-adrenomedullin antibody or an adrenomedullin antibody fragment binding to ADM or an ADM non—1g scaffold binding to ADM.

A person skilled in the art tands that anti-adrenomedullin (ADM) antibody or an anti“ adrenornedullin antibody fragment or DM nonwlg scaffold is synonymous t0 adrenomedullin (ADM) dy or an adrenornedullin antibody fragment or ADM non-1g scaffold and means anti—adrenomedullin (ADM) antibody or an anti—adrenomedullin antibody 1.0 fragment or anti—ADM non—lg scaffold binding to ADM, respectively.

Severe diseases are non-contagious diseases Whose al and social symptoms can have a severe impact on sufferers' abilities to live normal everyday life. In one embodiment severe disease is a life—threatening disease that bears a high risk of mortality caused by said severe e.

Said disease or condition may be selected from the group comprising severe infections as itis, Systemic inflammatory Response—Syndrom (SIRS), sepsis; other diseases as diabetes, cancer, acute and chronic ar diseases as sag. heart failure, myocardial infarction, stroke, atherosclerosis; shock as eg. septic shock and organ dysfunction as eg. kidney dysfunction, liver dysfunction, burnings, y, traumata.

In another embodiment said severe disease is an acute disease or condition as eg. meningitis, Systemic inflammatory Response—Syndrom (SIRS), sepsis, acute heart failure, myocardial infarction, ; shock as ag. septic shock and organ dysfunction as eg. kidney dysfunction, liver dysfunction, burnings, surgery, traumata. Shock may be septic shock or shock due to eg. burnings, surgery, traumata.

Especially useful is the antibody or fragment or scaffold according to the present ion for reducing the risk of mortality during SIRS, severe infection, sepsis and septic shock, tie. late phases of sepsis. This means that said anti—ADM antibody or anti—ADM antibody fragment binding to ADM or anti—ADM non—1g scaffold binding to ADM is used in therapy or prevention of severe ion as eg. meningitis, SIRS, , severe , shock as cg. septic shock. The person skilled the art understands that said anti-ADM antibody or anti~ ADM antibody fragment binding to ADM or anti—ADM nonalg scaffold binding to ADM may be administered systemically in patients having severe infection as eg. meningitis, SIRS, sepsis, severe sepsis, shock as cg. septic shock or may be administered ically in patients having the risk of getting a severe infection as tag. meningitis, SIRS, sepsis, severe sepsis, shock as ag. septic shock.

In the following clinical criteria for SIRS, sepsis, severe sepsis, septic shock will be defined. 1) Systemic inflammatory host response (SIRS) characterized by at least two of the following symptoms patients exhibit hypotension (mean arterial pressure is < 65 mm Hg) elevated serum lactate level being > 4 mmol/L blood glucose > 7.7 mmol/L (in e of diabetes) central venous pressure is not within the range 8—12 mm Hg urine output is < 0.5 mL 2; kg'1 x hr”I central venous (superior vena cava) oxygen saturation is < 70% or mixed venous < heart rate is > 90 beats/min temperature < 36°C or > 38°C respiratory rate > QO/min white cell count < 4 or > L (leucocytes); > 10% immature neutrophils 2) Sepsis Following at least two of the symptoms mentioned under 1), and additionally a clinical suspicion ofnew infection, being it: cough/sputum/chest pain nal istension/diarrhoea line infection endocarditis o dysuria o headache with neck stiffness - cellulitis/wound/joint infection 0 positive microbiology for any infection 3) Severe sepsis Provided that sepsis is manifested in patient, and additionally a clinical suspicion of any organ dysfunction, being it: 0 blood pressure systolic < 90/mean; < 65mmHG o e > 2 mincl/L - Bilirubine > 34nincl/L o urine output < 0.5 iankg/h for 2h 0 creatinine >177 umol/L o platelets < 9/L o SpOg > 90% unless 02 given 4) Septic shock At least one sign of end—organ dysfunction as mentioned under 3) is manifested. Septic shock is indicated, if there is refractory hypotension that does not respond to treatment and intravenous fluid administration alone is insufficient to maintain a patient's blood pressure from becoming hypotensive also provides for an administration of an anti—ADM antibody or an anti—ADM antibody fragment or an anti-ADM non—1g scaffold in ance with the present invention.

In accordance with the invention acute e or acute conditions may be selected from the group but are not limited to the group comprising severe infections as rag. meningitis, Systemic atory se—Syndrome (SIRS), or ; other diseases as diabetes, cancer, acute and chronic vascular diseases as cg. heart failure, myocardial infarction, stroke, atherosclerosis; shock as eg. septic shock and organ ction as eg. kidney dysfunction, WO 72510 liver ction, burnings, surgery, ta, poisoning, damages induced by chemotherapy.

Especially useful is the antibody or fragment or scaffold according to the present invention for reducing the risk of mortality during sepsis and septic shock, 15‘. e. late phases of .

In another embodiment said acute e or acute condition is selected from the group comprising meningitis, diabetes, cancer, acute and chronic vascular diseases as ag. heart failure, myocardial infarction, stroke, atherosclerosis; shock as ag. septic shock and organ dysfunction as e. g. kidney dysfunction, liver dysfunction, burnings, surgery, ta, poisoning, s induced by chemotherapy. rmore, in one embodiment of the invention an anti-Adrenomedullin (ADM) dy or an anti—adrenomedullin antibody fragment or an anti—ADM non—1g scaffold is monospecific.

Monospecitic anti-adrenomedullin (ADM) antibody or ecific anti-adrenomedullin antibody fragment or monospecific anti—ADM non-lg scaffold means that said dy or antibody fragment or non—lg scaffold binds to one specific region encompassing at least 5 amino acids Within the target ADM. ecific anti—Adrenornedullin (ADM) antibody or monospecific antiuadrenomeduilin dy fragment or monospecific anti—ADM non-lg scaffold are anti—adrenomedullin (ADM) antibodies or anti—adrenomedullin antibody fragments or anti—ADM non~1g scaffolds that all have affinity for the same antigen.

In another specific and red embodiment the anti—ADM antibody or the anti-ADM antibody fragment or anti—ADM non-lg scaffold binding to ADM is a monospecific antibody, antibody fi-aginent or non—lg scaffoid, respectively, whereby monospecific means that said antibody or antibody fragment or non—lg scaffold binds to one specific region encompassing at least 4 amino acids within the target ADM. Monospecific antibodies or nts or non-lg scaffolds according to the invention are antibodies or fragments or non—lg scaffolds that all have affinity for the same antigen. Monoclonal antibodies are monospecific, but monospecific antibodies may also be produced by other means than producing them from a common germ cell.

Said anti—ADM antibody or antibody fragment binding to ADM or non—lg scaffold binding to ADM may be a non—neutralizing anti—ADM antibody or antibody fragment binding to ADM or non—1g scaffold binding to ADM.

For other diseases blocking of ADM may be beneficial to a certain extent. However, it might also be detrimental if ADM is y neutralized as a. certain amount of ADM may be required for several physiological functions. In many reports it was emphasized that the stration of ADM may be beneficial in certain diseases. In contrast thereto in other reports ADM was reported as being life threatening when administered in certain conditions.

Thus, in a specific embodiment of the invention the anti—ADM antibody or anti—ADM antibody fragment or anti—ADM non—lg scaffold may be a non~neutralizing antibody/antibody fragment/non—Ig scaffold.

In a specific embodiment said anti—ADM antibody, anti—ADM antibody fragment or anti— ADM non—1g scaffold is a non-neutralizing antibody, fragment or non-lg scaffold. A neutralizing anti-ADM antibody, anti—ADM antibody fragment or anti—ADM non-1g scaffold would block the ivity of ADM to nearly 100%, to at least more than 90%, preferably to at least more than 95%.

In contrast, a utralizing anti—ADM antibody, or anti—ADM antibody nt or anti~ ADM non—lg ld blocks the bioactivity of ADM less than 100%, preferably to less than 95%, preferably to less than 90%, more preferred to less than 80 % and even more preferred to less than 50 %. This means that bioactivity of ADM is reduced to less than 100%, to 95 % or less but not more, to 90 0/0 or less but not more , to 80 % or less but not more to 50 % or less but not more This means that the residual bioactivity of ADM bound to the non~ neutralizing anti—ADM dy, or anti—ADM antibody fragment or anti-ADM non-lg scaffold would be more than 0%, preferably more than 5 %, preferably more than 10 % more preferred more than 20 %, more preferred more than 50 %.

In this t (a) molecu1e(s), being it an antibody, or an antibody nt or a non-1g scaffold with “non-neutralizing anti—ADM activity”, collectively termed here for simplicity as “non-neutralizing” anti-ADM antibody, antibody fragment, or non—1g scaffold, that ag. blocks the ivity ofADM to less than 80 %, is defined as ~ a molecule or molecules g to ADM, which upon addition to a culture of an eukaryotic cell line, which expresses functional human recombinant ADM receptor composed of CRLR (calcitonin receptor like or) and RAMP3 (receptor— ty modifying protein 3), reduces the amount of CAMP produced by the cell line through the action of parallel added human synthetic ADM peptide, wherein said added human synthetic ADM is added in an amount that in the absence of the non-neutralizing antibody to be analyzed, leads to half-maximal stimulation of CAMP synthesis, wherein the reduction of CAMP by said molecule(s) binding to ADM takes place to an extent, which is not more than 80%, even when the non- neutralizing molecule(s) binding to ADM to be analyzed is added in an , which is d more than the amount, which is needed to obtain the maximal reduction of CAMP synthesis obtainable with the non—neutralizing antibody to be analyzed.

The same definition applies to the other ranges; 95%, 90%, 50% etc.

As shown in Example 4 said DM antibody or antinADM antibody fragment or anti— ADM non—lg scaffold may reduce the mortality of a patient having a severe chronical or acute disease or acute condition when used as primary medication. For instance, in Sepsis patients, primary medication may be the treatment with antibiotics. Example 4 shows that the administration of said anti-ADM antibody or antibody fragment or non—1g scaffold may reduce mortality even if no fiarther measures are taken. This may be of l ance ag. in case of antibiotics ance or atibiiity against antibiotics. In other cases said anti— ADM antibody or anti-ADM antibody fragment or anti-ADM non—lg scaffold may be administered in combination with antibiotics. in a specific embodiment said anti—ADM antibody or antinADM antibody fragment or anti- ADM non—1g ld may be administered for treating said patient even if the patient has no signs yet of circulation problems, eg. low blood pressure, and f or if the patient has no signs of fluid imbalance yet or / if the patient has no signs yet of organ dysfunction. The administration of anti—ADM antibody or antiwADM antibody fragment or anti—ADM non—1g scaffold results in a ization of capillary blood flow by adjusting ADM to a physiological level and is thus influencing the e for said patient directly and reduces mortality in said patients. In accordance with the invention the expression “administration of said DM dy or anti-ADM antibody fragment or anti—ADM nonnig scaffold” denotes systemic administration. in one embodiment of the invention an anti—adrenomeduiiin (ADM) antibody or an anti~ADM dy fiagment or anti—ADM non-lg scaffold is to be used in combination with vasopressors ag. catecholamine wherein said combination is for use in therapy of a chronic or acute disease or acute condition of a patient for reducing the risk of mortality in said patient.

In one embodiment of the invention said patient having a chronic or acute e or condition is characterized by the need of said patient to get administration of vasopressors e.g. catecholamine administration.

Furthermore, in one embodiment of the ion an drenomedullin (ADM) antibody or an anti—adrenomedullin dy nt or an anti—ADM non-1g scaffold is to be used in combination With fluids administered intravenously, wherein said combination is for use in therapy of a patient having a chronic or acute disease or acute condition of a patient for reducing the risk of mortality in said patient. It should be emphasized that said intravenous administration of fluids means a systemic administration of fluids.

In one embodiment of the ion said patient having a chronic or acute disease or condition is characterized by the need of said patient to get intravenous fluids.

Subject matter of the invention in one specific embodiment is, thus, an anti—adrenomeduilin (ADM) antibody or an drenomedullin antibody fragment or anti—ADM non—1g ld for use in therapy of a patient in need of intravenous fluids.

In this regard, fluid balance is the balance of the input and the output of fluids in the body to allow metabolic processes to on. Dehydration is d as a i% or greater loss of body mass as a result of fluid loss. The three elements for assessing fluid e and hydration status are: clinical assessment, body weight and urine output; review fluid balance charts and review of blood chemistry. All this is very well known to a man skilled in the art (Alison Shepherd, Nursing Tomes 19.07.11/Voi 107 N0 28, pages 12 to 16).

Thus, in one embodiment a person in need of regulating the fluid balance, and thus in need of intravenous fluids is a person that has a 1% or greater loss of body mass as a result of fluid loss. The fluid balance may be assessed according to Scales and Pilsworth (2008) Nursing Standard 22:47, 50—57. For instance, normal urine output is in the range of 0.5 to 2 ml/kg of body weight per hour. The minimum acceptable urine output for a patient with normal renal function is 0.5 nil/kg per hour. All these rds may be used to assess whether a patient is in need of intravenous fluids, or synonymously in need for regulating the fluid balance and/or improving the fluid balance.

An antibody or fragment according to the present invention is a protein including one or more polypeptides substantially encoded by immunoglobulin genes that specifically binds an antigen. The recognized immunoglobulin genes e the kappa, lambda, alpha (IgA), gamma (IgGi, IgGg, IgGg, IgGa), delta (IgD), epsilon (IgE) and mu (IgM) constant region genes, as well as the myriad immunoglobulin variable region genes. Full—length immunoglobulin light chains are lly about 25 Kd or 214 amino acids in length. Full— length immunoglobulin heavy chains are generally about 50 Kd or 446 amino acid in length.

Light chains are encoded by a variable region gene at the NIB—terminus (about 110 amino acids in length) and a kappa or lambda constant region gene at the COOH--terminus. Heavy chains are similarly d by a variable region gene (about 116 amino acids in length) and one of the other constant region genes.

The basic structural unit of an antibody is generally a tetramer that consists of two identical pairs of immunoglobulin chains, each pair having one light and one heavy chain. In each pair, the light and heavy chain variable regions bind to an antigen, and the constant regions mediate effector functions. lmmunoglobulins also exist in a variety of other forms including, for e, Fv, Fab, and (Fab'h, as well as bifunctional hybrid antibodies and single chains (sag, ecchia et ah, Eur. J. Immunol. 17:105,1987; Huston et‘ at, Proc. Natl. Acad. Sci.

USA, 85,-5879—5883, 1988; Bird 2: al., Science 242:423—426, 1988; Hood et at, Immunology, Benjamin, N.Y., 2nd ed., 1984; Hunkapiller and Hood, Nature 323:15~l6,l986).

An immunoglobulin light or heavy chain variable region includes a framework region interrupted by three hypervariable regions, also called complementarity determining regions (CDR’s) (see, Sequences of Proteins of logical Interest, E. Kabat et at, US.

Department of Health and Human Services, 1983). As noted above, the CDRs are primarily responsible for binding to an epitope of an antigen. An immune complex is an antibody, such as a monoclonal antibody, chimeric antibody, humanized dy or human antibody, or functional dy nt, specifically bound to the antigen.

Chimeric antibodies are antibodies whose light and heavy chain genes have been constructed, lly by genetic ering, from innnunoglobulin variable and constant region genes belonging to different s. For example, the variable segments of the genes from a mouse monoclonal antibody can be joined to human constant segments, such as kappa and gamma 1 or gamma 3. In one example, a therapeutic chimeric antibody is thus a hybrid protein composed of the variable or antigenubinding domain from a mouse antibody and the constant or or domain from a human antibody, although other mammalian species can be used, or the variable region can be ed by molecular techniques. s of making chimeric antibodies are well known in the art, e.g., see U.S. Patent No. 5,807,715. A ized" globulin is an immunoglobulin including a human framework region and one or more CDRs from a non—human (such as a mouse, rat, or synthetic) immunoglobulin. The nou— human immunoglobulin providing the CDRs is termed a " and the human immuneglobulin providing the framework is termed an "acceptor." In one embodiment, all the CDRs are from the donor immunoglobulin in a humanized globulin. Constant regions need not be present, but if they are, they must be substantially identical to human immunoglobulin constant s, 118., at least about 85—90%, such as about 95% or more identical. Hence, all parts of a humanized immunoglobulin, except possibly the CDRs, are IS substantially cal to corresponding parts of natural human immunoglobulin sequences. A "humanized antibody" is an antibody sing a humanized light chain and a humanized heavy chain immunoglobulin. A humanized antibody binds to the same antigen as the donor antibody that provides the CDRs. The acceptor framework of a humanized immunoglobulin or antibody may have a limited number of substitutions by amino acids taken from the donor framework. Humanized or other monoclonal antibodies can have additional conservative amino acid substitutions which have substantially no effect on antigen binding or other globulin functions. Exemplary conservative substitutions are those such as gly, ala; val, ile, leu; asp, gin; asn, gln; ser, thr; lys, arg‘, and phe, tyr. Humanized immunoglobulins can be constructed by means of genetic engineering (rag, see US. Patent No. 089). A human antibody is an antibody wherein the light and heavy chain genes are of human .

Human antibodies can be generated using methods known in the art. Human antibodies can be produced by immortalizing a human B cell secreting the antibody of interest. Immortalization can be accomplished, for example, by EBV ion or by fusing a human B cell with a myeloma or hybridoma cell to produce a triorna cell. Human antibodies can also be produced by phage display methods (see, cg, Dower et (11., PCT Publication No. WO9l/l727l; McCafferty ex (11., PCT Publication No. WO92/001047; and Winter, PCT ation No.

W092/20791), or selected from a human combinatorial monoclonal antibody library (see the Morphosys website). Human antibodies can also be prepared by using transgenic animals carrying a human immunoglobulin gene (for example, see Lonberg at 511., PCT PublicatiOn No. WO93/12227; and Kucherlapati, PCT Publication No. W0 91/10741).

Thus, the anti—ADM dy may have the formats known in the art. Examples are human antibodies, monoclonal antibodies, humanized antibodies, chimeric antibodies, CDR-grafied antibodies. In a preferred embodiment antibodies according to the present invention are recombinantly ed antibodies as ag. IgG, a typical full—length immunoglobulin, or antibody nts containing at least the F—variable domain of heavy and/or light chain as ag. ally coupled antibodies (fragment antigen binding) including but not d to Fab-fragments including Fab minibodies, single chain Fab antibody, monovalent Fab antibody with epitope tags, ag. Fab-VSSXZ; bivalent Fab (mini—antibody) dimerized with the CH3 domain; bivalent Fab or multivaient Fab, 9.g. formed via muitimerization with the aid of a logous domain, ag. via dimerization of dHLX s,e.g. Fab—dHLX-FSXZ; Hawk—fragments, scFv—fragments, multimerized multivalent or/and multispecific scFv~ fragments, bivalent and/or bispecific diabodies, BITE® (bispecific T—cell engager), trifunctional antibodies, polyvalent antibodies, tag. from a different class than G; single- domain antibodies, rag. nanobodies derived from camelid or fish immunoglobulinesand numerous others.

In addition to DM antibodies other biopolymer scaffolds are well known in the art to complex a target molecule and have been used for the tion of highiy target specific biopolymers. Examples are aptamers, spiegelmers, anticalins and conotoxins. For illustration of antibody formats please see Fig. la, lb and lo.

In a preferred embodiment the anti—ADM antibody format 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 format is selected from the group comprising scFab nt, Fab nt, scFv fragment and bioavailability optimized conjugates thereof, such as PEGylated fragments. One of the most preferred formats is the scFab format.

Non~lg scaffolds may be protein lds and may be used as antibody mimics as they are capable to bind to ligands or nes. Non—1g scaffolds may be selected from the group comprising tetranectin—based non—lg scaffolds (ag. described in US 2010/0028995), fibronectin scaffolds (cg. described in EP 1266 025; lipocalinubased scaffolds ((e.g. described in ); ubiquitin scaffolds (cg. described in ), transferrin scaffolds (cg. described in US 2004/0023334), protein A lds (tag. described in EP 2231860), ankyrin repeat based scaffolds (e.g. described in ), microproteins preferably microproteins forming a cysteine knot) scaffolds (cg. described in EP 2314308), Fyn SH3 domain based scaffolds (cg. described in ) EGFR-A—domain based scaffolds (cg. bed in ) and Kunitz domain based scaffolds (cg. described in EP 1941867).

In one embodiment of the invention anti-ADM antibodies according to the present invention may be produced as outlined in Example 1 by synthesizing fragments of ADM as antigens.

Thereafter, binder to said fragments are fied using the below described s or other methods as known in the art.

A Balb/c mouse was immunized with lOOug ADM Peptide~BSA—Conjugate at day 0 and l4 (emulsified in 100ul complete Freund’s adjuvant) and SOug at day 21 and 28 (in IOOul incomplete Freund’s adjuvant). Three days before the fusion experiment was perfonned, the animal received 50ug of the conjugate dissolved in 100m saline, given as one intraperitoueal and one intra—venous injection.

Spenocytes from the immunized mouse and cells of the myeloma cell line SPZ/O were fused with lml 50% polyethylene glycol for 30s at 37°C. After g, the cells were seeded in 96~well cell culture plates. Hybrid clones were selected by growing in HAT medium [RPMI 1640 culture medium supplemented with 20% fetal calf serum and HAT—Supplement]. After two weeks the HAT medium is replaced with HT Medium for three es followed by returning to the normal cell culture medium.

The cell culture supernatants were primary screened for antigen specific IgG antibodies three weeks after fusion. The positive tested ultures were transferred into 24—well plates for ation. After retesting, the selected cultures were cloned and recloned using the limiting—dilution technique and the isotypes were deteimined. (see also Lane, RD. (1985). A short-duration polyethylene glycol fusion que for sing production of monoclonal antibody—secreting hybridomas. J. l. Meth. 81: 223-228; Ziegler, B. at 96) Glutamate decarboxylase (GAD) is not detectable on the e of rat islet cells examined by cytofluorometry and complement—dependent antibody—mediated xicity of monoclonal GAD antibodies, Horm. Metab. Res. 28: 11—15).

Antibodies may be produced by means of phage display according to the following procedure: The human naive antibody gene libraries HAL7/8 were used for the isolation of recombinant single chain F—Variable domains (scFv) against medullin peptide. The antibody gene libraries were ed with a panning strategy comprising the use of peptides ning a biotin tag linked via two different spacers to the medullin peptide sequence. A mix of panning rounds using non—specifically bound antigen and streptavidin bound antigen were used to minimize background of non—specific binders. The eluted phages from the third round of panning have been used for the generation of monoclonal scFv expressing E.coli strains.

Supernatant from the cultivation of these clonal strains has been directly used for an antigen ELISA testing (see also Hust, M., Meyer, T., ch, B., Rfilker, T., Thie, H., El—Ghezal, A, Kirsch, M.I., te, M., Helmsing, S., Meier, D., mann, T., Diibel, S., 2011. A human scFV dy generation pipeline for proteome research. Journal of Biotechnology 152, 159—170; Schiitte, M., Thullier, P., Pelat, T., Wezler, X., Rosenstock, P., Hinz, D., Kirsch, M.I.,Hasenberg, M., Frank, R., Schinmann, T., Gunzer, M., Hust, M., Diibel, 3., 2009. Identification of a putative Crf splice variant and generation of recombinant antibodies for the specific detection of Aspergillus fumigatus. PLoS One 4, e6625) Humanization ofmurine antibodies may be conducted according to the following procedure: For zation of an antibody of murine origin the antibody sequence is analyzed for the structural interaction of framework regions (FR) with the complementary determining regions (CDR) and the antigen. Based on structural modeling an appropriate PR of human origin is selected and the murine CDR sequences are transplanted into the human FR. Variations in the amino acid ce of the CDRS or FRs may be introduced to regain structural interactions, which were abolished by the species switch for the FR sequences. This recovery of structural interactions may be achieved by random ch using phage display libraries or via directed approach guided by molecular modeling. (Almagro JC, Fransson J 2008.

Humanization of antibodies. Front . 2008 Jan l;13: 1619—33.) In a preferred embodiment the ADM dy format 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 red embodiment the antibody format is selected from the group comprising scFab fragment, Fab fragment, scFv fragment and bioavailability optimized conjugates thereof, such as PEGylated fragments. One of the most preferred formats is scFab format.

In another preferred embodiment, the anti—ADM antibody, anti—ADM antibody fragment, or anti~ADM non-lg scaffold is a full length antibody, antibody fragment, or non—lg scaffold.

In a preferred embodiment the anti—adrenomedullin antibody or an anti—adrenomedullin antibody fragment or anti—ADM non—1g scaffold is directed to and can bind to an epitope of at least 5 amino acids in length contained in ADM.

In a more preferred embodiment the anti—adrenomedullin antibody or an anti—adrenornedullin antibody fragment or anti-ADM non—Ig ld is directed to and can bind to an epitope of at least 4 amino acids in length contained in ADM.

In one specific embodiment of the invention the drenomedullin (ADM) antibody or anti— ADM antibody fragment binding to adrenomedullin or DM non-Ig scaffold g to adrenorneduilin is provided for use in therapy or prevention of an acute e or acute condition of a patient wherein said antibody or nt or scaffold is not ADM—binding— Protein—l (complement factor H).

In one specific embodiment of the invention the anti—Adrenorneduliin (ADM) antibody or anti~ADM antibody fragment binding to medullin or anti—ADM non—lg scaffold binding to adrenomeduliin is provided for use in therapy or prevention of an acute disease or acute condition of a patient wherein said antibody or antibody nt or non-1g scaffold binds to a region of preferably at least 4, or at least 5 amino acids within the sequence of aa 1— 42 of mature human ADM: SEQ ID No.: 24 NFQGLRSFGCRFGTCTVQKLAHQIYQFTDKDKDNVA.

In one specific embodiment of the invention the anti—Adrenomedullin (ADM) dy or anti-ADM dy fragment binding to adrenomedullin or anti~ADM non—1g scaffold binding to adrenomednllin is ed for use in therapy or prevention of an acute disease or acute condition ofa patient wherein said antibody or nt or scaffold binds to a region of preferably at least 4, or at least 5 amino acids within the sequence of aa 1—21 of mature human ADM: SEQ ID No.: 23 YRQSMNNFQGLRSFGCRFGTC.

In a preferred ment of the present invention said anti—ADM antibody or an anti— adrenomedullin antibody fragment or anti—ADM non-1g scaffold binds to a region or epitope ofADM that is located in the inal part (aa 1—21) of adrenomeduilin, (see Fig. 2).

In another preferred embodiment said anti-ADM—antibody or an anti-adrenomedullin antibody fragment or anti-ADM non-1g scaffold recognizes and binds to a region or e within amino acids 1-14 (SEQ ID NO: 26) of adrenomedullin; that means to the N—terrninal part (aa 1—14) of adrenomedullin. In another preferred embodiment said anti—ADM—antibody or an anti—adrenomeduilin antibody fragment or DM non—1g scaffold recognizes and binds to a region or epitope within amino acids 1—10 of adrenornedullin (SEQ ID NO: 27); that means to the N—terminal part (aa 1-10) of medullin. aa 1—14 ofADM YRQSMNNFQGLRSF (SEQ ID NO: 26) aa 1-10 ofADM YRQSMNNFQG (SEQ ID NO: 27) In another preferred embodiment said anti-ADM antibody or an anti—adrenomednllin antibody fragment or anti»ADM non—1g scaffold recognizes and binds to a region or epitope Within amino acids 1-6 of adrenomedullin (SEQ ID NO: 28); that means to the N-tenninal part (aa 1~ 6) of adrenonieduilin. As above stated said region or epitope comprises preferably at least 4 or at least 5 amino acids in length. aa 1—6 ofADM YRQSMN (SEQ ID NO: 28) in another preferred embodiment said anti-ADM antibody or an anti—adrenomedullin antibody fragment or anti—ADM non—1g scaffold recognizes and binds to the N—terrninal end (aal) of adrenomedullin. N—terrninal end means that the amino acid 1, that is “Y” of SEQ ID No. 21 or 23; is mandatory for antibody binding. The antibody or nt or scaffold would neither bind N—terininal extended nor N~terminal modified Adrenornedullin nor N—terrninal ed adrenomedullin. This means in another preferred embodiment said anti—ADM-antibody or an anti—adrenomedullin dy fragment or anti-ADM non—lg scaffold binds only to a region IO within the sequence of mature ADM if the inal end of ADM is free. In said embodiment the anti—ADM antibody or anti—adrenomedulh'n antibody fragment or non-1g scaffold would not bind to a region within the sequence of mature ADM if said sequence is e. g. comprised within pro—ADM.

For the sake of clarity the numbers in brackets for specific regions of ADM like “bl-terminal part (aa 1‘21)” is understood by a person skilled in the art that the N—terminal part of ADM consists of amino acids 1—21 of the mature ADM sequence.

In another specific embodiment pursuant to the invention the herein ed anti—ADM antibody or DM antibody fragment or anti~ADM non—1g scaffold does not bind to the C—terminal portion of ADM, tie. the aa 43 —~ 52 ofADM (SEQ ID NO: 25) PQGYmNHZ (SEQ ID N0225) In one specific embodiment it is preferred to use an anti—ADM antibody or an anti— adrenomedullin dy fragment or anti—ADM non~Ig scaffold according to the present invention, wherein said anti~adrenomedullin antibody or said anti~adrenomedullin antibody nt or non—lg scaffold is an ADM stabilizing antibody or an adrenomedullin izing antibody fragment or an adrenomedullin stabilizing non-1g scaffold that enhances the half life (tug; half retention time) of adrenomedullin in serum, blood, plasma at least 10 %, preferably at least 50 %, more preferably >50 %, most preferably >100%.

The half life (half retention time) of ADM may be determined in human plasma in absence and presence of an ADM stabilizing antibody or an adrenomedullin stabilizing antibody nt or an adrenornedullin stabilizing non—lg scaffold, respectively, using an immunoassay for the fication ofADM.

The following steps may be conducted: — ADM may be diluted in human citrate plasma in absence and presence of an ADM stabilizing antibody or an adrenomedullin stabilizing antibody fragment or an adrenomedullin izing non~lg ld, respectively, and may be incubated at 24 °C — Aliquots are taken at selected time points (ag. within 24 hours) and degradation of ADM may be stopped in said aliquots by freezing at ~20 °C — The quantity of ADM may be determined by an hADM assay directly, if the selected assay is not influenced by the stabilizing dy. Alternatively, the aliquot may be treated with denaturing agents (like HCl) and, after clearing the sample (eg. by centrifugation) the pH can be neutralized and the ADM—quantified by an ADM immunoassay. Alternatively, non-immunoassay technologies (ag. rpHPLC) can be used for ADM—quantification.

~ The half life of ADM is calculated for ADM incubated in absence and presence of an ADM stabilizing antibody or an adrenomedullin stabilizing antibody fragment or an adrenomedullin stabilizing non—1g scaffold, respectively, n The enhancement of half life is calculated for the stabilized ADM in comparison to ADM that has been incubated in absence of an ADM stabilizing dy or an adrenomednllin stabilizing dy fragment or an adrenomedullin stabilizing non—lg scaffold.

A twofold increase of the half life of ADM is an enhancement ofhalf life of 100%.

Half Life (half retention time) is defined as the period over which the concentration of a specified chemical or drug takes to fall to half its baseline concentration in the ed fluid or blood.

An assay that may be used for the determination of the Half life (half retention time) of adrenomedullin in serum, blood, plasma is described in Example 3.

In a preferred embodiment said anti—ADM antibody, anti—ADM antibody nt or anti- ADM non—1g scaffold is a non-neutralizing antibody, fragment or scaffold. A neutralizing anti—ADM antibody, anti—ADM antibody fragment or anti—ADM non-lg scaffold would block the bioactivity of ADM to nearly 100%, to at least more than 90%, preferably to at least more than 95%. In other words this means that said non~neutralizing anti-ADM antibody, anti» ADM antibody fragment or anti~ADM non—lg scaffold blocks the bioactivity of ADM to less than 100 %, preferably less than 95% preferably less than 90%. In an embodiment n said utralizing anti-ADM antibody, anti—ADM antibody fragment or anti—ADM non-lg scaffold blocks the bioactivity of ADM to less than 95% an anti—ADM antibody, anti—ADM antibody fragment or DM non—lg scaffold that would block the bioactivity of ADM to more than 95 % would be outside of the scope of said embodiment. This means in one embodiment that the bioaetiyity is reduced to 95 % or less but not more, preferably to 90 % or less, more ably to 80 % or less, more preferably to 50 % or less but not more. in one embodiment of the ion the non-neutralizing antibody is an antibody binding to a region of at least 5 amino acids within the sequence of aa 1—42 of mature human ADM(SEQ ID NO: 24), preferably within the sequence aa 1—32 of mature human ADM (SEQ ID NO: 29), or an antibody binding to a region of at least 5 amino acids Within the sequence of aa 1— 40 of mature e ADM (SEQ ID NO: 30), ably Within the sequence aa l~31 of mature murine ADM (SEQ ID NO: 31). ln another preferred embodiment of the invention the non—neutralizing antibody is an antibody binding to a region of at least 4 amino acids Within the sequence of aa 142 of mature human ADM (SEQ ID NO: 24), preferably within the sequence aa 1—32 of mature human ADM (SEQ ID NO: 29), or an antibody binding to a region of at least 4 amino acids within the sequence of aa 1—40 of mature murine ADM (SEQ ID NO: 30), preferably Within the sequence aa 1—31 of mature murine ADM (SEQ ID NO: 31). aa 1~32 human mature human ADM YRQSMNNFQGLRSFG-CRFGTCTVQKLAHQIYQ (SEQ ID NO: 29) aa 1-40 mature murine ADM YRQSMNQGSRSNGCRFGTCTFQKLAHQIYQLTDKDKDGMA (SEQ ID NO: 30) aa 1-31 mature murine ADM QGSRSNGCRFGTCTFQKLAHQIYQL (SEQ ID NO: 31) In a specific embodiment according to the present invention an non—neutralizing anti—ADM dy or an anti—adrenomedullin antibody fragment or ADM non—lg scaffold is used, wherein said anti—ADM antibody or an drenomedullin antibody fragment blocks the bioactivity of ADM to less than 80 %, preferably less than 50% (of baseline values). It has been understood that said limited blocking of the ivity (meaning reduction of the bioactivity) of ADM occurs even at excess concentration of the antibody, nt or scaffold, g an excess of the antibody, fragment or scaffold in relation to ADM. Said limited blocking is an intrinsic property of the ADM binder itself in said specific embodiment. This means that said antibody, fragment or scaffold has a l inhibition of 80% or 50% respectively. In a preferred embodiment said anti-ADM dy, anti—ADM antibody fragment or anti—ADM non—lg scaffold would block the bioactivity / reduce the bioactivity of anti-ADM to at least 5 %. The stated above means that approximately 20% or 50% or even 95% residual ADM bioactivity remains present, respectively.

Thus, in ance with the t invention the provided anti—ADM antibodies, anti—ADM antibody fi'agments, and anti—ADM non-lg scaffolds do not neutralize the respective ADM bioactivity.

The bioactivity is defined as the effect that a substance takes on a living organism or tissue or organ or functional unit in vivo or in vitro (eg. in an assay) after its interaction. In case of ADM bioactivity this may be the effect of ADM in a human recombinant Adrenomedullin receptor cAMP functional assay. Thus, ing to the present invention bioactivity is defined via an Adrenomedullin receptor CAMP functional assay. The following steps may be performed in order to determine the bioactivity ofADM in such an assay: — Dose response curves are performed with ADM in said human recombinant Adrenornedullin receptor cAMP functional assay.

— The ADM-concentration of half-maximal CAMP stimulation may be calculated.

, At constant halfumaximal cAMP~stimulating ADM—concentrations dose response curves (up to lOOug/ml final concentration) are med by an ADM stabilizing antibody or an adrenomedullin stabilizing antibody fragment or an adrenomedullin stabilizing non—lg scaffold, respectively.

A maximal inhibition in said ADM bioassay of 50% means that said anti~ADM antibody or said antiuadrenornedullin antibody fragment or said anti—adrenomedullin nonnlg scaffold, respectively, blocks the bioactivity of ADM to 50% of baseline values. A l inhibition in said ADM bioassay of 80% means that said anti—ADM antibody or said anti- adrenornedullin antibody fragment or said anti-adrenomedullin non—1g scaffold, respectively, blocks the bioactivity 0f ADM to 80%. This is in the sense of blocking the ADM bioactivity to not more than 80%. This means imately 20% al ADM bioactivity remains r, by the t specification and in the above context the expression “blocks the bioactivity of ADM” in relation to the herein disclosed anti-ADM antibodies, antiuADM antibody fragments, and anti-ADM non—lg lds should be tood as mere sing the bioactivity of ADM from 100% to 20% remaining ADM ivity at maximum, preferably decreasing the ADM bioactivity from 100% to 50% remaining ADM bioactivity; but in any case there is ADM bioactivity remaining that can be determined as detailed above.

The bioaetivity of ADM may be determined in a human recombinant Adrenomedullin or CAMP fianctional assay (Adrenomedullin Bioassay) according to Example 2.

In a preferred embodiment a modulating antibody or a modulating anti-adrenomedullin antibody fragment or a modulating anti—adrenomedullin non—lg scaffold is used in therapy or prevention of a chronic or acute disease or acute condition of a patient for stabilizing the circulation, in ular stabilizing the systemic circulation.

Such a modulating anti-ADM antibody or a modulating anti—adrenomeduliin antibody fragment or a modulating anti—adrenomedullin non~Ig scaffold may be especialiy useful in the treatment of sepsis. A modulating antibody or a modulating anti—adrenomedullin antibody fragment or a modulating anti—adrenomedullin non—lg scaffold enhances the bioactivity of ADM in the early phase of sepsis and reduces the ng effects of ADM in the late phase of sepsis.

A “modulating” anti—ADM antibody or a modulating anti—adrenomedullin antibody fragment or a modulating anti—adrenomedullin non—1g scaffold is an antibody or an anti—adrenomedullin antibody fragment or non-lg scaffold that enhances the half life (111/2 half ion time) of adrenornedullin in serum, blood, plasma at least 10 9/9, preferably at least, 50 %, more preferably >50 %, m0st preferably >100% and blocks the bioactivity of ADM to less than 80 %, preferably less than 50 “/0 and said anti—ADM antibody, anti—ADM antibody fragment or anti-ADM non—lg scaffold would block the bioactivity of ADM to at least 5 %. These values related to ife and blocking of bioactivity have to be understood in relation to the before- mentioned assays in order to determine these values. This is in the sense of ng the ADM bioactivity of not more than 80% or not more than 50%, respectively.

Such a modulating anti—ADM antibody or a modulating anti—adrenomedullin antibody nt or a modulating anti~adrenomedullin non~lg scaffold offers the advantage that the dosing of the administration is facilitated. The combination of partially blocking or partially reducing Adrenomedullin bioactivity and increase of the in vivo half life (increasing the Adrenomedullin bioactivity) leads to beneficial simplicity of anti—adrenomeduilin antibody or an anti-adrenomedullin dy fragment or anti-adrenomeduliin non—lg scaffold dosing. In a situation of excess of endogenous medullin (maximal stimulation, late sepsis phase, shock, hypodynamic phase) the activity lowering effect is the major impact of the antibody or fragment or scaffold, limiting the (negative) effect of Adrenomedullin. In case of low or normal endogenous Adrenomedullin concentrations, the biological effect of anti~ adrenomedullin dy or an anti~adrenornedullin antibody fragment or anti—ADM non—1g scaffold is a combination of lowering (by partially blocking) and increase by increasing the Adrenomedullin half life. If the half life effect is stronger than the blocking effect, the biological net activity of nous Adrenomedullin is beneficially increased in early phases of Sepsis (low Adrenomedullin, hyperdynamic phase). Thus, the non—neutralizing and ting anti—adrenomedullin antibody or drenomedullin antibody fragment or anti— adrenomednllin non—1g scaffold acts like an ADM bioactivity buffer in order to keep the bioactivity ofADM within a certain logical range.

Thus, the dosing of the antibody/fragment/scaffold in eg. sepsis may be selected fiom an excessive concentration, because both Sepsis phases (early and late) benefit from excessive anti~ADM antibody or an drenomedullin antibody fragment or anti—ADM non—lg ld treatment in case of a ting effect. ive means: The anti—adrenomedullin WO 72510 antibody or an anti—adrenomedullin antibody fragment or anti-ADM non—lg scaffold concentration is higher than endogenous Adrenomedullin during late phase (shock) of ag.

Sepsis. This means, in case of a modulating anti—ADM dy or modulating anti—ADM antibody fragment or ting anti-ADM non—lg scaffold dosing in sepsis may be as follows: The concentration of Adrenornedullin in septic shock is 226+/—66 frnol/ml (Nishio er al., "Increased plasma concentrations of adrenomedullin correlate with tion of vascular tone in ts with septic shock", Crit Care Med. 1997, 25(6):953—7), an equimolar concentration of antibody or fragment or scaffold is 42.5ug/l blood, (based on 6 1 blood volume / 80kg body weight) 3.2ug/kg body weight. Excess means at least double (mean) septic shock Adrenomedullin concentration, at least > 3ug anti-adrenomedullin antibody or an antitadrenomedullin antibody fragment or anti~ADM non—lg scaffold / kg body weight, preferred at least 6.4ug antinadrenomedullin antibody or an anti—adrenomedullin antibody fragment anti-ADM non—1g scaffold fkg body weight. red > long / kg, more preferred >20ug/kg, most preferred >100ug Anti—adrenomedullin antibody or an anti—adrenomcdullin dy fragment or DM non—lg Scaffold / kg body weight.

This may apply to other severe and acute conditions than septic shock as well.

In a Specific embodiment of the invention the antibody is a monoclonal antibody or a nt thereof. In one embodiment of the invention the anti—ADM dy or the anti« ADM antibody fragment is a human or humanized antibody or derived rom. In one specific embodiment one or more (inurine) CDR’s are grafted into a human antibody or antibody fragment.

Subject matter of the present invention in one aspect is a human CDR—grafted antibody or antibody fragment thereof that binds to ADM, n the human CDRngrafted antibody or antibody fragment thereof comprises an antibody heavy chain (H chain) comprising SEQ ID NO:1 GYTFSRYW SEQ ID NO: 2 ILPGSGST and/or SEQ ID NO: 3 TEGYEYDGFDY and/or further comprises an antibody light chain (L chain) comprising: SEQ ID NO: 4 QSWYSNGNTY SEQ ID NO: 5 and/or SEQ ID NO: 6 FQGSHIPYT.

In one specific embodiment of the invention subject matter of the present invention is a human monoclonal antibody that binds to ADM or an antibody fragment thereof that binds to ADM wherein the heavy chain ses at least one CDR selected from the group comprising: SEQ ID NO: 1 GYTFSRYW SEQ ID NO: 2 ILPGSGST SEQ ID NO: 3 TEGYEYDGFDY and wherein the light chain comprises at least one CDR ed from the group comprising: 2012/072929 SEQ ID No: 4 QSIVYSNGNTY SEQ ID NO: 5 SEQ ID NO: 6 FQGSHIPYT.

In a more specific embodiment of the invention subject matter of the ion is a human monoclonal antibody that binds to ADM or an antibody fragment thereof that binds to ADM wherein the heavy chain comprises the sequences SEQ ID NO: 1 GYTFSRYW SEQ ID NO: 2 ILPGSGST SEQ ID NO: 3 TEGYEYDGFDY and wherein the light chain comprises the sequences SEQ ID NO: 4 QSIVYSNGNTY SEQ ID NO: 5 SEQ ID No: 6 FQGSHIPYT.

In a very specific embodiment the anti—ADM dy has a sequence selected from the group comprising: SEQ ID NO 7, 8,9, 10, 11, 12,13 and 14.

Such a modulating antibody or an adrenomedullin antibody fragment offers the advantage that the dosing of the stration is facilitated. The ation of partially blocking or partially reducing medullin bioactivity and increase of the in vivo half life (increasing the Adrenomedullin bioactivity) leads to beneficial simplificity of Adrenomedullin antibody or an adrenomedullin antibody fragment dosing. In a situation of excess of endogenous Adrenomedullin (maximal stimulation, late sepsis phase, shock, hypodynamic phase) the activity ng effect is the major impact of the dy or fragment, ng the (negative) effect of Adrenomedullin. In case of low or normal endogenous Adrenomedullin concentrations, the biological effect of Adrenomedullin antibody or an adrenomedullin antibody fragment is a combination of lowering (by partially blocking) and increase by increasing the Adrenomedullin half life. If the half life effect is stronger than the blocking effect, the biological activity of endogenous Adrenomedullin is beneficially increased in early phases of Sepsis (low Adrenomedullin, hyperdynamic phase). The dosing of antibody can be selected from an excessive concentration, e both Sepsis phases (early and late) benefit from excessive antibody or an adrenomedullin antibody fragment treatment in case of a modulating effect. ive means: The anti Adrenomedullin antibody or an adrenomedullin antibody fragment concentration is higher than endogenous medullin during late phase (shock) of Sepsis. This means, in case of a modulating antibody or modulating fragment dosing may be as follows: The concentration of medullin in septic shock is 226+f—66 fmol/ml (Nishio er a1, 1997), an equimolar concentration of antibody or fragment is 42,5ug/1 blood, (bei 61/ 80kg) 3,2"ug/kg body weight. Excess means at least double (mean) septic shock Adrenomedullin concentration, at least > 3ug Adrenomedullin antibody or an adrenomedullin antibody fragment / kg body weight, preferred at least 6,4ug Adrenomedullin antibody or an adrenomedullin antibody nt Ikg body weight. Preferred > lOug / kg, more preferred >20ug/kg, most preferred >100ug Adrenomedullin antibody or an adrenomedullin antibody nt / kg body weight.

The anti—ADM dy or anti—adrenomedullin antibody fragment or anti-ADM non—lg scaffold according to the present invention exhibits an affinity towards human ADM in such WO 72510 that affinity constant is greater than 10‘7 M, preferred 10‘8 M, preferred affinity is greater than '9 M, most preferred higher than 10'10 M. A person skilled in the art knows that it may be considered to compensate lower affinity by applying a higher dose of compounds and this measure would not lead out—of—the—scope of the invention. The y constants may be determined according to the method as described in e 1.

The antibodies or fragments according to the present invention can be used in combinations with other agents, as e. g. with the so—called ADM—binding protein, for the uses as described therein.

In a red embodiment the antibody or the antibody fragment is used for reducing the risk of mortality during said chronic or acute disease of a t wherein said disease is selected from the group sing sepsis, diabetes, cancer, acute and chronic vascular diseases as heart failure, shock as ag. septic chock and organ dysfunction as eg. kidney dysfunction.

Especially useful is the antibody or fragment according to the present invention for reducing the risk of mortality during sepsis and septic shock, is. late phases of sepsis.

In a preferred embodiment the anti-ADM antibody or the anti-ADM antibody fragment or anti—ADM nondg scaffold is used for reducing the risk of mortality during said chronic or acute disease or acute condition of a patient. c or acute disease or acute condition according to the present invention may be a disease or condition selected from the group comprising severe infections as ag. meningitis, Systemic inflammatory Response~Syndrom , sepsis; other diseases as diabetes, cancer, acute and chronic vascular diseases as e.g. heart failure, myocardial infarction, , atherosclerosis; shock as eg. septic shock and organ dysfunction as ag. kidney dysfunction, liver dysfunction, acute myocardial infarction, acute stroke, polytraumatic events, gs, surgery, poisoning, damages by chemotherapy. Especially useful is the antibody or nt or ld according to the present invention for ng the risk of mortality during sepsis and septic shock, tie. late phases of sepsis.

In one embodiment the anti—ADM antibody or an anti-adrenomeduilin antibody nt or anti-ADM non—lg scaffold is used in therapy of a chronic or acute disease or acute condition of a patient according to the present invention wherein said patient is an lCU patient. In another embodiment the anti—ADM antibody or an anti—adrenomedullin antibody fragment or anti—ADM non-lg scaffold is used in therapy of a chronic or acute disease of a patient according to the present invention wherein said patient is ally ill. Critically ill means that a patient is having a disease or state in which death is possible or imminent.

Subject of the present invention is r an anti-ADM antibody or DM antibody fiagment or antiwADM non—lg scaffold for use in therapy of a chronic or acute disease or acute ion of a patient according to the present invention wherein said antibody or IO fragment or scaffold is to be used in ation of ADM g protein. ADM g protein is also naturally present in the circulation of said patient.

It should be emphasized that the term ADM binding protein also denotes ADMw‘oinding- protein—1 (complement factor H), which however is not a non—neutralizing and modulating anti—ADM dy, anti—ADM antibody fragment, or anti-ADM non—1g scaffold as in accordance with the invention.

Subject of the present invention is r an anti—ADM antibody or an antiuadrenomedullin antibody fragment or anti-ADM non—lg scaffold for use in therapy of a chronic or acute disease or acute condition of a patient according to the present invention wherein said antibody or fragment or scaffold is to be used in combination with further active ingredients.

Subject matter of the invention is also an anti-adrenomedullin (ADM) dy or an anti— adrenomednllin antibody fragment or an anti~ADM non-lg scaffold to be used in combination with a another y medicament wherein said ation is for use in therapy of a chronic or acute disease or acute condition of a patient for stabilizing the circulation of said patient, in particular for stabilizing the systemic circulation of said patient.

Primary medicament means a medicament that acts against the primary cause of said disease or condition. Said primary medicament may be antibiotics in case of infections.

Surprisingly, it has been shown by the present invention that an anti—ADM antibody or an anti-adrenomedullin antibody fragment or anti—ADM non—lg scaffold may act as a y medicament for ng the risk of mortality in said severe diseases and conditions. Example 4, in particular Table 4, shows that t any other treatment than treatment with anti—ADM antibody or an anti-adrenomedullin antibody fragment or anti—ADM non—1g scaffold the risk of mortality may be considerably reduced tag. in sepsis. This means in case of sepsis or other severe infections anti—ADM antibody or an drenomeduilin antibody fragment or anti— ADM non—1g scaffold may be administered t any r medication in order to reduce mortality in said ts. This may be of speciai ance if there is a resistance against the other primary medication like antibiotics resistance or in case if there is an intolerance against the other primary medication as ag. an antibiotics intolerance. In such cases the treatment with anti—ADM antibody or an anti—adrenorneduliin antibody fragment or anti-ADM non—1g scaffold as only primary medication may be considered.

Secondary medication is a medication that improves the condition of the patient in a supportive way as ag. stabilizing the circulation in a patient having an acute disease or ion. In ular, this is in the sense of stabilizing the systemic circulation in a patient having an acute disease or condition.

Such tive measures may be administration of vasopressors ag. of oiamine administration and / or fiuid administration.

Surprisingly, it has been found that the treatment with an anti—ADM antibody or an anti- adrenomedullin antibody fragment or anti—ADM non—1g scaffold may be applied as primary medication in order to reduce mortality in said diseases and ions.

Thus, in a specific embodiment of the before mentioned combinations said combinations are to be used in combination with vasopressors ag. catecholarnine wherein said further combination is for use in therapy of a chronic or acute disease or condition of a t for reducing the risk of mortality.

In one embodiment of the invention said patient having a chronic or acute disease or chronic condition is characterized by the need of the patient to get stration of vasopressors tag. catechoiamine administration.

In another embodiment of the ion said anti—ADM antibody or an anti-adrenomeduilin antibody fragment or anti-ADM non—1g scaffold may be applied in combination with antibiotics.

Throughout the specification, it should be emphasized that application or administration of the anti—ADM antibody or an anti—adrenomedullin antibody fragment or DM non—lg scaffold according to the invention is preferably a ic application / administration.

Subject matter of the invention in one specific embodiment is, thus, an anti—adrenomedullin (ADM) antibody or an anti—adrenomedullin antibody fragment or an anti—ADM nonmlg scaffold to be used in combination with ADM binding protein and/or further active ingredients for use in y of a patient in need of a treatment of vasopressors e.g. catecholamine treatment.

In a specific ment of the above mentioned combinations said combinations are to be used in combination with fluids administered intravenously, wherein said combination is for use in therapy of a chronic or acute e or condition of a patient for ng the risk of mortality. 1t shouid be emphasized that application or administration of the anti-ADM antibody or an anti—adrenomedullin antibody fragment or anti-ADM non-1g scaffold according to the invention is preferably a systemic application / administration.

In one embodiment of the invention said patient having a chronic or acute e or acute condition is characterized by the need of the patient to get intravenous fluids.

Subject matter of the invention in one specific embodiment is, thus, an Anti—adrenomedullin (ADM) antibody or an drenomedullin dy nt or anti—ADM non—1g scaffold in combination with ADM binding protein and/or further active ingredients for use in therapy of a t in need of intravenous fluids.

Said anti-ADM antibody or an anti—adrenomedullin antibody fragment or anti-ADM non~lg scaffold or combinations thereof with ADM binding n and/or further active ingredients may be used in combination with vasopressors e.g. catecholamine and/or with fluids administered intravenously for use in a of a chronic or acute disease or acute ion. It should be emphasized that application or administration of the anti-ADM antibody or an anti— adrenomedullin antibody fragment or anti-ADM non—lg scaffold according to the invention is preferably a systemic application / administration.

But as ed before in r specific embodiment said anti—ADM antibody or anti-ADM antibody nt or anti—ADM non~lg ld may be administered for ng said patient even if the patient has no signs of circulation problems, eg. low blood pressure, and / or if the patient has no signs of fluid imbalance or / if the patient has no signs of organ dysfunction.

The administration of anti—ADM antibody or antimADM antibody fragment or anti—ADM non— Ig scaffold results in a normalization of capillary blood flow by adjusting anti—ADM to a physiological level and is thus influencing the outcome for said patient directly and reduces mortality in said patients. Thus, in this another specific embodiment, the patient may not be in need of a treatment of vasopressors eg. catecholamine treatment and f or may not be in need of intravenous fluids and/or may not have any signs of organ dysfimction, as eg. kidney or liver dysfunction.

Subject matter of the invention is also an anti-ADM antibody or an drenomedullin dy fragment or anti—ADM non-lg scaffold according to the t invention to be used in combination with TNF-alpha—antibodies. TNF-alpha—antibodies are commercially available for the treatment of ts. t of the present invention is further a ceutical formulation comprising an antibody or fragment or scaffold according to the present invention. t of the present invention is r a pharmaceutical formulation according to the present invention wherein said pharmaceutical formulation is a solution, preferably a ready to—use solution.

Said pharmaceutical formulation may be administered intra—muscular. Said pharmaceutical formulation may be administered intra-vascular. Said pharmaceutical formulation may be administered via infusion.

In another embodiment subject of the present ion is further a pharmaceutical formulation according to the present invention wherein said pharmaceutical formulation is in a dried state to be reconstituted before use.

In r embodiment subject of the t invention is further a pharmaceutical formulation according to the present invention wherein said pharmaceutical formulation is in a freeze—dried state.

It should be emphasized that the ceutical formulation in accordance with the invention as may be administered intra—muscular, intra-vascular, or via infusion is preferably administered systemically to a patient for reducing the risk of mortality in a patient having a chronic or acute disease or acute condition.

Therefore, in another embodiment of the present invention the pharmaceutical ation according to the present invention is to be administered systemically to a patient for reducing the risk of mortality in a patient having a chronic or acute disease or acute condition.

In a c embodiment the pharmaceutical formulations according to the invention are intended for use in therapy or prevention of SIRS, a severe infection, sepsis, shock tag. septic shock in a patient for the reduction of the mortality risk for said t, wherein said pharmaceutical formulation is to be administered systemically.

Further embodiments within the scope of the present invention are set out below: 1. medullin ADM antibody or an adrenomedullin antibody fragment for use in therapy of a chronic or acute disease of a patient for the regulation of liquid balance. 2. ADM antibody or an adrenomedullin antibody fragment according to claim 1 wherein the antibody format is ed from the group comprising Fv nt, scFV fragment, Fab fragment, scFab nt, (Fab)2 fragment and scFvch Fusion protein. 3. ADM antibody or an adrenomedullin antibody fragment according claim 1 or 2 wherein said antibody or fragment binds to the N-terminal part (aa 1—21) of adrenomedullin. 4. ADM antibody or an adrenomedullin antibody fragment according to any of claims 1 to 3, wherein said dy or fragment recognizes and binds to the N—terminal end (aal) of adrenomedullin.

. ADM antibody or an adrenomedullin antibody fragment ing to any of claims 1 to 4, wherein said antibody or fragment is an ADM stabilizing dy or ADM stabilizing a antibody fragment that enhances the tug half ion time of adrenomedullin in serum, blood, plasma at. least 10 %, preferably at least 50 %, more preferably >50 %, most preferably >100 %.

ADM antibody or an adrenomeduliin antibody fragment according to any of claims 1 to 5, wherein said antibody or fragment blocks the bioactivity ofADM to less than 80 %, preferably less than 50%.

ADM antibody or an adrenomedullin antibody fragment for use in y of a chronic or acute disease of a patient according to any of claims 1 to 6 wherein said disease is selected from the group comprising , diabetis, cancer, heart failure, shock and kidney dysfunction.

ADM antibody or an adrenomedullin antibody fragment for use in therapy of a chronic or acute disease of a patient according to any of claims 1 to 7 wherein said patient is an ICU patient.

ADM antibody or an medullin antibody nt for use in therapy of a chronic or acute disease of a patient according to any of claims 1 to 7 wherein said antibody or fragment is a modulating antibody or fragment that enhances the rm half retention time of medullin in serum, blood, plasma at least 10 0/0, preferably at least 50 %, more preferably >50 %, most preferably >100 % and that blocks the bioactivity ofADM to less than 80 %, preferably less than 50%. 10. Pharmaceutical formulation comprising an antibody or fragment according to any of claims 1 to 9. ll. ceutical formulation according to claim 10 wherein said pharmaceutical ation is a solution, preferably a ready~to—use solution. 12. ceutical formulation according to claim 10 wherein said pharmaceutical formulation is in a mdried state. 13. ceutical formulation according to any of claims 10 to 11, wherein said pharmaceutical formulation is administered intramuscular. 14. Pharmaceutical formulation according to any of claims 10 to ll, wherein said pharmaceutical formulation is administered intra—vascular.

. Pharmaceutical formulation according to claim 14, wherein said pharmaceutical formulation is administered via infusion. r embodiments within the scope of the present invention are set out below: Adrenomedullin ADM antibody or an adrenomedullin antibody fragment an ADM non-lg scaffold for use in therapy of a chronic or acute disease or acute condition of a patient for the regulation of fluid balance.

ADM dy or an adrenomedullin antibody fragment or ADM non-1G scaffold according to claim 1 wherein said ADM antibody or an adrenoniedullin antibody fragment or ADM non-1G scaffold is a non~neutralizing ADM antibody or a non— neutralizing adrenornedullin antibody fragment or a non—neutralizing ADM noanG scaffold.

Adrenomedullin ADM antibody or an adrenomedullin antibody fragment or an ADM non—lg scaffold for use in therapy of a chronic or acute disease or acute ion according to claim 1 or 2 for ting or reducing edema in said t.

ADM dy or an medullin antibody fragment or ADM non-IG scaffold according to any of claims 1 to 3 wherein the antibody format is selected from the group comprising Fv fragment, scFv fragment, Fab fragment, scFab fragment, (Fab)2 fragment and scFv—Fc Fusion protein.

ADM antibody or an adrenomedullin antibody fragment or ADM non~lG scaffold according to any of claims 1 to in said antibody or fragment or scaffold binds to the Nmterminal part (aa 1—21) of adrenomedullin. 6. ADM antibody or an adrenomednllin antibody fragment or ADM non—1G scaffold according to any of claims 1 to 5, wherein said antibody or fragment scaffold recognizes and binds to the N—terminal end (aal) of adrenomedullin. 7. ADM antibody or an adrenomedullin antibody fragment or ADM non—IG scaffold according to any of claims 1 to 6, wherein said antibody or fragment or scaffold is an ADM izing antibody or ADM stabilizing antibody fragment or ADM izing non-1G ld that enhances the half life (ti/2 half retention time) of rnedullin in serum, blood, plasma at least 10 0/0, preferably at least 50 %, more preferably >50 %, most preferably >100 %. 8. ADM antibody or an adrenomedullin dy fragment or ADM non—1G scaffold according to any of claims 1 to 7, wherein said antibody or fragment blocks the bioactivity ofADM to less than 80 %, preferably less than 50%. 9. ADM antibody or an adrenomedullin antibody fragment or ADM non—1G scaffold for use in therapy of a chronic or acute disease of a patient according to any of claims 1 to 8 n said disease is selected from the group comprising SIRS, sepsis, diabetis, cancer, heart failure, shock and kidney dysfunction . ADM antibody or an adrenomedullin antibody fragment according to any of claims 1 to 9, wherein said antibody or fragment is a human onal antibody or fragment that binds to ADM or an antibody fragment thereof n the heavy chain comprises the sequences SEQ ID NO: 1 GYTFSRYW SEQ ID NO: 2 ILPGSGST SEQ ID NO: 3 TEGYEYDGFDY and wherein the light chain ses the sequences SEQ ID N024 QSIVYSNGNTY SEQ ID NO: 5 SEQ ID NO: 6 FQGSHIPYT. 11. A human monoclonal antibody or fragment that binds to ADM or an antibody nt thereof according to claim 10 wherein said antibody or fragment comprises a sequence selected from the group comprising : SEQ ID NO: 7 (AM—VH—C) QVQLQQSGAELMKPGASVKISCKATGYTFSRYWIEWVKQRPGHGLEWIGEILP YNEKFKGKATITADTSSNTAYMQLSSLTSEDSAVYYCTEGYEYDGFD YWGQGTTLTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKR VEPKHHHHHH SEQ ID NO: 8 (AM-VH1) QVQLVQSGAEVKKPGSSVKVSCKASGYTFSRYWISWVRQAPGQGLEWMGRIL PGSGSTNYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCTEGYEYDGFD YWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKR VEPKHHHHHH SEQ ID NO: 9 (AM—VH2-E40) SGAEVKKPGSSVKVSCKASGYTFSRYWIEWVRQAPGQGLEWMGRIL PGSGSTNYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCTEGYEYDGFD YWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKR VEPKHHHHHH SEQ ID NO: 10 (AM~VH3—T26—E55) QVQLVQSGAEVKKPGSSVKVSCKATGYTFSRYWISWVRQAPGQGLEWMGEIL NYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCTEGYEYDGFD YWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVDKR VEPKHHHHHH SEQ ID NO: 11 (AM—VH4—T26-E40—E55) QVQLVQSGAEVKKPGSSVKVSCKATGYTFSRYWIEWVRQAPGQGLEWMGEIL PGSGSTNYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCTEGYEYDGFD YWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN GVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKR VEPKHHHHHH SEQ ID NO: 12 (AM—VL-C) DVLLSQTPLSLPVSLGDQATISCRSSQSIVYSNGNTYLEWYLQKPGQSPKLLIYR VSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDLGVYYCFQGSHIPYTFGGGTKLE IKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNS QESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGE SEQ ID NO: 13 (AM-VLI) DVVMTQSPLSLPVTLGQPASISCRSSQSWYSNGNTYLNWFQQRPGQSPRRLIYR VSNRDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFQGSHIPYTFGQGTKL EIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGN SQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRG SEQ ID NO: 14 (AM-VLZ—E40) DVVMTQSPLSLPVTLGQPASISCRSSQSIVYSNGNTYLEWFQQRPGQSPRRLIYR VSNRDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFQGSHIPYTFGQGTKL EIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGN SQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRG 12. ADM antibody or an adrenomednilin antibody fragment or ADM non~IG scaffold for use in therapy of a chronic or acute disease of a patient according to any of claims 1 to 9 wherein said patient is an ICU patient. 13. ADM antibody or an adrenomedullin antibody fragment or ADM non~IG scaffold for use in therapy of a chronic or acute e of a patient according to any of claims 1 to 12 wherein said antibody or nt or ld is a modulating antibody or fragment or scaffold that enhances the half life (tug half retention time) of adrenomedullin in serum, blood, plasma at least 10 %, preferably at least 50 %, more preferably >50 %, most preferably >100 % and that blocks the bioactivity of ADM to less than 80 %, preferably less than 50%. 14. ADM antibody or an adrenomedullin antibody fragment or ADM non—1G scaffold for use in therapy of a chronic or acute e of a patient according to any of the claims I to 13 to be used in combination with catecholamine and/ or fluids administered enously.

. ADM antibody or medullin antibody fragment or ADM non-1G scaffold for use in therapy of a chronic or acute disease of a patient according to any of the claims 1 to 13 or a combination according to claim 12 to be used in combination with ADM binding protein and/0r further active ingredients. 16. Pharmaceutical formulation sing an antibody or fragment or scaffold according to any of claims 1 to 15. 17. Pharmaceutical formulation according to claim 16 n said pharmaceutical formulation is a solution, ably a ready-to—use solution. 18. Pharmaceutical formulation according to claim 16 wherein said ceutical formulation is in a freeze-dried state. 19. Phannaceutical formulation according to any of claims 16 to 17, wherein said pharmaceutical formulation is administered intra-muscular.

. Pharmaceutical formulation according to any of claims 16 to 17, wherein said pharmaceutical formulation is administered intra—vascular. 21. ceutical formulation according to claim 20, wherein said pharmaceutical formulation is administered Via infusion.

Further embodiments within the scope of the present invention are set out below: Adrenomedullin (ADM) antibody or an adrenomedullin antibody fragment for use in therapy of a chronic or acute disease of a patient for stabilizing the circulation.

ADM antibody or an adrenomedullin antibody fragment according to claim 1 n said antibody or fragment reduces the olamine requirement of said patient.

ADM antibody or an adrenomedullin dy nt according to claim 1 or 2 wherein the antibody format is selected from the group comprising FV fragment, scFV fragment, Fab fragment, scFab fragment, (Fab)2 fragment and scFV—Fc Fusion protein.

ADM antibody or an adrenomedullin antibody fragment according to any of claims 1 to 3 wherein said antibody or fragment binds to the N—terminal part (aa l»21) of adrenomedullin.

. ADM dy or an adrenomedullin antibody fragment according to any of claims 1 to 4, wherein said antibody or fragment recognizes and binds to the N—terminal end (aal) of medullin.

ADM dy or an adrenomedullin antibody fragment according to any of claims I to , wherein said antibody or fragment is an ADM stabilizing antibody that enhances the t1/2 half retention time of adrenomedullin in serum, blood, plasma at least 10 %, ably at least, 50 %, more ably > 50 “/05 most preferably >100 %.

ADM antibody or an adrenomedullin antibody fragment according to any of claims 1 to 6, wherein said antibody or fragment blocks the bioactivity of ADM to less than 80 %, preferably less than 50 %.

ADM antibody or an adrenomedullin antibody fragment according to any of claims 1 to 7, wherein said antibody or fragment is a modulating ADM antibody or a modulating adrenomedullin antibody fragment that enhances the tl/2 half retention time of adrenomedullin in serum, blood, plasma at least 10 %, preferably at least, 50 %, more preferably > 50 %, most preferably >100 % and that blocks the bioactivity of ADM to less than 80 %, ably less than 50 %: ADM antibody or an adrenornedullin antibody fragment for use in therapy of a chronic or acute disease of a patient according to any of the claims 1 to 8 wherein said disease is selected from the group comprising sepsis, diabetis, cancer, acute and c vascular diseases as e.g. heart failure, shock as ag. septic shock and organ dysfunction as eg. kidney dysfunction.

. Pharmaceutical ation comprising an antibody according to any of claims I to 9. ll. Pharmaceutical formulation according to claim 10 wherein said pharmaceutical formulation is a solution, preferably a ready—to~use solution 12. Pharmaceutical formulation according to claim 10 wherein said pharmaceutical formulation is in a freeze—dried state. 13. Pharmaceutical formulation according to any of claims 10 to 11, wherein said pharmaceutical formulation is administered intramuscular. l4. Pharmaceutical formulation according to any of claims 10 to ll, wherein said pharmaceutical formulation is stered intrauvascular.

. Pharmaceutical ation according to claim 14, wherein said pharmaceutical formulation is administered via infusion.

Further embodiments within the scope of the present invention are set out below: 1. Adrenomedullin (ADM) antibody or an adrenomedullin antibody fragment or an ADM non-IG scaffold for use in therapy of a chronic or acute disease or condition of a patient for stabilizing the circulation. 2. ADM antibody or an adrenomedullin antibody fragment or ADM non—1G scaffold according to claim 1 n said antibody or fragment or scaffold reduces the vasopressor ement, ag. catecholamine ement of said patient. 3. ADM antibody or an adrenomedullin antibody fragment or ADM non—1G scaffold according to claim 1 or 2 wherein said ADM dy or an rnedullin antibody fragment or ADM non—1G scaffold is a non—neutralizing ADM dy or a non— neutralizing adrenomedullin antibody fragment or a non-neutralizing ADM non—1G scaffold. 4. ADM antibody or an adrenomeduilin antibody fragment according to any of claims 1 to 3 wherein the antibody format is selected from the group comprising Fv fragment, scFV fragment, Fab fragment, scFab nt, (Fab)2 fragment and scFv-Fc Fusion protein. 5. ADM dy or an adrenomedullin antibody fragment or ADM non—1G scaffold according to any of claims 1 to 4 wherein said antibody or fragment or scaffold binds to the N—terminal part (aa 1—21) of adrenomedullin. 2012/072929 ADM antibody or an medullin antibody fragment or ADM non—1G scaffold ing to any of claims 1 to 5, wherein said antibody or fragment or scaffold recognizes and binds to the inal end (aal) of adrenomedullin.

ADM antibody or an adrenomedullin antibody fragment or ADM non-1G scaffold according to any of claims 1 to 6, wherein said dy or fragment or scaffold is an ADM stabilizing antibody or nt or scaffold that enhances the half life (ti/2 half retention time) of adrenomedullin in serum, blood, plasma at least 10 “/13, preferably at least, 50 %, more preferably > 50 %, most preferably >100 %.

ADM antibody or an adrenomedullin antibody fragment or ADM non—1G scaffold according to any of claims 1 to 7, wherein said antibody or fragment or scaffold blocks the bioactivity ofADM to less than 80 %, preferably less than 50 %.

ADM antibody or an adrenomedullin dy fragment or ADM non-IG scaffold according to any of claims 1 to 8, wherein said antibody or fragment or scaffold is a modulating ADM antibody or a modulating adrenomedullin antibody fi'agment or scaffold that enhances the half life (ti/2 half retention time) of adrenomedullin in serum, blood, plasma at least 10 %, preferably at least, 50 0/0, more preferably > 50 %, most preferably >100 0/0 and that blocks the bioactivity ofADM to less than 80 %, ably less than 50 %: . ADM antibody or an adrenomedullin antibody fragment according to any of claims 1 to 9, wherein said antibody or fragment is a human onal antibody or fragment that binds to ADM or an antibody fragment thereof wherein the heavy chain comprises the sequences SEQ in NO: 1 GYTFSRYW SEQ ID NO: 2 lLPGSGST SEQ ID NO: 3 TEGYEYDGFDY and wherein the Eight chain comprises the ces SEQ ID N024 QSWYSNGNTY SEQ ID NO: 5 SEQ ID NO: 6 FQGSHIPYT. 11. A human monoclonal antibody or fragment that binds to ADM or an antibody fragment thereof according to claim 10 wherein said antibody or fragment ses 3 Sequence selected from the group comprising : SEQ ID NO: 7 (AM-VH—C) QVQLQQSGAELMKPGASVKISCKATGYTFSRYWIEWVKQRPGHGLEWIGEILP GSGSTNYNEKFKGKATITADTSSNTAYMQLSSLTSEDSAVYYCTEGYEYDGFD YWGQGTTLTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKR VEPKHHHHHH SEQ ID NO: 8 (AM-VH1) SGAEVKKPGSSVKVSCKASGYTFSRYWISWVRQAPGQGLEWMGRIL PGSGSTNYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCTEGYEYDGFD YWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKR VEPKHHHHHH SEQ ID NO: 9 (AM—VH2-E40) QVQLVQSGAEVKKPGSSVKVSCKASGYTFSRYWIEWVRQAPGQGLEWMGRIL NYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCTEGYEYDGFD YWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKR VEPKHHI-IHHH SEQ ID NO: 10 (AM—VH3~T26—E55) QVQLVQSGAEVKKPGSSVKVSCKATGYTFSRYWISWVRQAPGQGLEWMGEIL PGSGSTNYAQKFQGRVTITADBSTSTAYMELSSLRSEDTAVYYCTEGYEYDGFD YWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKR VEPKHHHHHH SEQ ID NO: 11 (AM-VH4-T26-E40—E55) QVQLVQSGAEVKKPGSSVKVSCKATGYTFSRYWIEWVRQAPGQGLEWMGEIL PGSGSTNYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCTEGYEYDGFD YWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEP‘VTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKR VEPKHHHHHH SEQ ID NO: 12 (AM—VL~C) TPLSLPVSLGDQATISCRSSQSIVYSNGNTYLEWYLQKPGQSPKLLIYR VSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDLGVYYCFQGSHIPYTFGGGTKLE APSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNS QESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGE SEQ ID NO: 13 (AM—VLI) DVVMTQSPLSLPVTLGQPASISCRSSQSIVYSNGNTYLNWFQQRPGQSPRRLIY RVSNRDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFQGSHIPYTFGQGT KLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQ SGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKS FNRGEC SEQ ID NO: 14 (AM—VLZ-E40) SPLSLPVTLGQPASISCRSSQSIVYSNGNTYLEWFQQRPGQSPRRLIY RVSNRDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFQGSHIPYTFGQGT KLEIKRTVAAPSVFIFPPsDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQ SGNSQESVTBQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKS FNRGEC 12. ADM antibody or an medullin antibody fragment or ADM non—1G scaffold for use in therapy of a chronic or acute e of a t according to any of the claims 1 to 11 wherein said disease is selected from the group comprising SIRS, sepsis, diabetis, cancer, acute and chronic vascular diseases as eg. heart failure, shock as eg. septic shock and organ dysfunction as 62g. kidney dysfunction. 13. ADM antibody or an adrenomedullin antibody fragment or ADM non-1G ld for use in therapy of a c or acute e of a patient according to any of the claims 1 to 12 to be used in combination with catecholamine and/ or fluids administered intravenously. 14. ADM antibody or adrenomedullin antibody fragment or ADM non—1G scaffold for use 2O in therapy of a chronic or acute disease of a patient according to any of the claims 1 to 13 or a combination according to claim 10 to be used in combination with ADM binding protein and/or further active ingredients. . ceutical formulation comprising an antibody or fragment or non-IG scaffold according to any of claims 1 to 14. 16. Pharmaceutical formulation according to claim 15 wherein said pharmaceutical formulation is a soiution, preferably a ready—to~use solution. 17. Pharmaceutical formulation according to claim 15 wherein said pharmaceutical formulation is in a freeze—dried state. 18. Pharmaceutical formulation according to any of claims 15 to 16, wherein said pharmaceutical formulation is administered muscular. 19. Phannaceutical ation according to any of claims 14 to 16, wherein said pharmaceutical formulation is administered intra-vascular.

. Pharmaceutical formulation according to claim 16, wherein said pharmaceutical ation is administered via infusion.

Further embodiments within the scope of the present invention are set out below: 1. Adrenomedullin antibody or an adrenomedullin antibody fragment for use in a treatment of a c or acute disease n said antibody or said fragment is an ADM stabilizing antibody or fragment that enhances the tug half retention time of adrenomedullin in serum, blood, plasma at least 10 %, preferably at least, 50 “/0, more ably >50 %, most preferably 100 % and/or wherein said antibody blocks the bioactivity of ADM to less than 80 %, preferably to less than 50 %. 2. Adrenomedullin antibody or an adrenomedullin antibody fragment for use in a treatment of a chronic or acute disease n said antibody or said fragment is a modulating ADM antibody or fragment that enhances the tug half retention time of adrenomedullin in serum, blood, plasma at least 10 %, preferably at least, 50 %, more preferably >50 %, most preferably 100 % and that blocks the bioactivity of ADM to less than 80 %, preferably to less than 50 %. 3. Adrenomedullin antibody or an adrenomedullin antibody fragment for use in a treatment of a chronic or acute disease according to claim 1 or 2, wherein said dy or fragment binds to the N—terminal part (aa 1—21) of adrenomedullin. 4. Adrenomedullin antibody or an adrenornedullin antibody fragment for use in a treatment of a chronic or acute e n said antibody or said fragment according to claim 3 binds to the N-terminal end of adrenomedullin.

Adrenomedullin antibody or an adrenomeduliin antibody fragment for use in use in a treatment of a chronic or acute disease according to any of claims 1 to 4, wherein said antibody or said fragment is an ADM stabilizing antibody or fragment that enhances the t; 12 half retention time of adrenomeduliin in serum, blood, plasma at least 10 %, preferably at least, 50 %, more preferably >50 %, most ably 100 0/0.

Adrenomedullin antibody or an adrenomedullin antibody fragment for use in a treatment of a chronic or acute disease ing to any of claims 1 to 5, wherein said antibody or said fragment blocks the bioactivity of ADM to less than 80 %, preferably to less than 50 %.

Adrenomedullin antibody or an adrenomedullin antibody fragment according to any of the claims 1 to 6 for use in a treatment of a chronic or acute disease wherein said disease is selected from the group sing SIRS, sepsis, septic shock, diabetis, cancer, heart failure, shock, organ failure, kidney dysfunction, acute liquid ance, and low blood pl‘CSSIll‘fi.

Adrenomedullin dy or an adrenomeduilin antibody fragment according to any of the claims 1 to 7 for use in a treatment of a chronic or acute disease wherein said disease is septic shock or .

Adrenomeduliin antibody or an adrenomeduliin antibody fragment for use in a treatment of a c or acute disease ing to any of the claims 1 to 8 wherein said antibody or fragment regulates the liquid balance of said patient. 10. medullin antibody or an adrenomeduliin antibody fragment for use in a ent of a chronic or acute disease according to any of the claims 1 to 9 wherein said antibody or fragment used for prevention of organ dysfunction or organ failure. 11. Adrenomedullin antibody or an adrenomedullin antibody fragment for use in a treatment of a chrome or acute disease according to claim 10 wherein said dy or fragment is used for prevention of kidney dysfunction or kidney failure. 12. Adrenomedullin (ADM) antibody or an adrenomedullin antibody fragment for use in a treatment of a chronic or acute disease in a patient according to claims 1 to 11 wherein said antibody or nt is used for izing the circulation. 13. ADM antibody or an adrenomedullin antibody fragment for use in a treatment of a chronic or acute disease in a patient according to claim 12 n said antibody or fragment s the catecholamine requirement of said patient. 14. ADM antibody or an adrenornedullin antibody fragment for use in a treatment of a chronic or acute disease in a patient ing to any of claims 1 to 13 for the reduction of the mortality risk for said patient.

. ADM dy or an adrenomedullin antibody fragment for use in a treatment of a chronic or acute disease in a patient according to any of claims 1 to 14 n said antibody or fragment may be administered in a dose of at least 3 ug / Kg body . 16. Pharmaceutical composition comprising an antibody or fragment according to any of claims 1 to 15.

Further embodiments Within the scope of the present invention are set out below: Adrenomeduilin antibody or an medullin dy fragment or ADM non—lg scaffold wherein said antibody or said fragment or scaffold is a nonmneutralizing antibody.

Adrenomedullin antibody or an adrenomedullin antibody fragment or ADM non-lg scaffold wherein said antibody or said fragment or scaffold is an ADM stabilizing antibody or fragment or scaffold that enhances the half life (tug half retention time) of adrenomedullin in serum, blood, plasma at least 10 %, preferably at least 50 %, more preferably >50 %, most preferably 100 % and/or wherein said antibody or fragment or scaffold blocks the bioactivity of ADM to less than 80 %, preferably to less than 50 %. 3. Adrenomedullin antibody or an adrenomeduilin antibody fragment or ADM non—1g scaffold wherein said antibody or said fragment is a modulating ADM dy or fragment or scaffold that enhances the half life (ll/2 half retention time) of adrenomedullin in serum, blood, plasma at least 10 %, preferably at least, 50 %, more preferably >50 0/0, most preferably 100 % and that blocks the bioactivity of ADM to less than 80 ”/0, preferably to less than 50 %. 4. Adrenomedullin antibody or an adrenomedullin antibody fragment or ADM non—lg scaffold ing to claim 1 or 2, wherein said dy or fragment or scaffold binds to the ninal part (aa 1—21) of adrenomedullin, . Adrenornedullin antibody or an adrenomedullin antibody fragment or ADM non—lg scaffold wherein said antibody or said fragment or scaffold according to claim 3 binds to the inal end of adrenomedullin. 6. medullin antibody or an adrenomedullin antibody fragment ADM non-1g scaffold according to any of claims 1 to 4, wherein said dy or said fragment or said scaffold is an ADM stabilizing antibody or nt that enhances the tm half retention time of adrenomedullin in serum, blood, plasma at least 10 %, ably at least, 50 0/0, more preferably >50 %, most preferably 100 %. 7. Adrenomedullin antibody or an adrenomedullin antibody fragment or ADM non-1g scaffold according to any of the claims 1 to 6 for use as an active pharmaceutical substance. 8. Adrenomedullin antibody or an adrenomedullin antibody fragment ADM non—lg scaffold according to any of the claims 1 to 7 for use in a treatment of a chronic or acute disease or acute condition wherein said disease or condition is selected from the group comprising severe infections as 6.5;. meningitis, systemic inflammatory Response- Syndrome (SlRS,) sepsis; other diseases as diabetes, cancer, acute and chronic vascular es as eg. heart failure, myocardial infarction, stroke, atherosclerosis; shock as rag.

WO 72510 Septic shock and organ dysfunction as e. g. kidney dysfunction, liver dysfunction, burnings, surgery, trauinata.

Adrenomedullin antibody or an adrenomedullin antibody fragment or ADM non—lg ld according to any of the claims 1 to 8 for use in a treatment of a chronic or acute disease or acute condition wherein said e is septic shock or sepsis.

. ADM antibody or an adrenomedullin antibody fragment according to any of claims 1 to 9, wherein said antibody or fragment is a human monoclonal antibody or fragment that binds to ADM or an antibody fragment thereof wherein the heavy chain comprises at least one of the sequences : SEQ ID NO: 1 GYTFSRYW SEQ ID NO: 2 ILPGSGST SEQ ID NO: 3 TEGYEYDGFDY And/0r wherein the light chain comprises the at ieast one of the sequences SEQ ID NO:4 QSIVYSNGNTY SEQ ID NO: 5 SEQ ID NO: 6 WO 72510 FQGSHIPYT.

A human monoclonal antibody or fragment that binds to ADM or an antibody fragment f according to claim 10 wherein said antibody or fragment comprises a sequence selected from the group comprising: SEQ ID NO: 7 —C) QVQLQQSGAELMKPGASVKISCKATGYTFSRYWIEWVKQRPGHGLEWIGEILP GSGSTNYNEKFKGKATITADTSSNTAYMQLSSLTSEDSAVYYCTEGYEYDGFD YWGQGTTLTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW SGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVDKR VEPKHHHHHH SEQ ID NO: 8 (AM—VH1) QVQLVQSGAEVKKPGSSVKVSCKASGYTFSRYWISWVRQAPGQGLEWMGRIL PGSGSTNYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCTEGYEYDGFD YWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVDKR HHHH SEQ ID NO: 9 (AM-VH2—E40) QVQLVQSGAEVKKPGSSVKVSCKASGYTFSRYWIEWVRQAPGQGLEWMGRIL PGSGSTNYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCTEGYEYDGFD YWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKR VEPKHHHHHH SEQ ID NO: 10 (AM~VH3-T26—E55) QVQLVQSGAEVKKPGSSVKVSCKATGYTFSRYWISWVRQAPGQGLEWMGEIL PGSGSTNYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCTEGYEYDGFD YWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKR VEPKHHHHHH SEQ ID NO: 11 (AM-VH4uT26-E40—ESS) SGAEVKKPGSSVKVSCKATGYTFSRYWIEWVRQAPGQGLEWMGEIL PGSGSTNYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCTEGYEYDGFD TVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKR VEPKHHHHHH SEQ ID NO: 12 (AM—VL—C) DVLLSQTPLSLPVSLGDQATISCRSSQSIVYSNGNTYLEWYLQKPGQSPKLLIYR VSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDLGVYYCFQGSHIPYTFGGGTKLE IKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNS QESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGE SEQ ID NO: 13 (AM-VLI) DVVMTQSPLSLPVTLGQPASISCRSSQSIVYSNGNTYLNWFQQRPGQSPRRLIYR VSNRDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFQGSHIPYTFGQGTKL EIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGN SQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRG SEQ ID NO: 14 (AM—VLZ—E40) DWMTQSPLSLPVTLGQPASISCRSSQSIVYSNGNTYLEWFQQRPGQSPRRLIYR VSNRDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFQGSHIPYTFGQGTKL EIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGN SQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRG 12. Adrenomeduliin antibody or an meduliin antibody fragment or ADM non—1g scaffold according to any of the claims 1 to 11 for reguiating the fluid balance in a patient having a chronic or acute disease or acute condition. 13. Adrenomedullin antibody or an adrenomeduflin antibody fragment or ADM non-1g scaffold according to any of the claims 1 to 11 for ting or reducing organ ction or organ failure in a patient having in a chronic or acute disease or acute condition. 14. Adrenomedullin antibody or an adrenomedullin antibody fragment or ADM non—1g scaffold according to claim 13 n organ is kidney or liver.

. Adrenomedullin (ADM) antibody or an adrenomedullin antibody fragment or ADM non—1g scaffold according to claims 1 to 14 for stabilizing the circulation in a patient having a c or acute disease or acute condition. 16. ADM antibody or an adrenomedullin antibody fragment or ADM non—lg scaffold for use in a treatment of a chronic or acute disease in a patient according to claim 15 wherein said antibody or fragment reduces the catecholamine requirement of said patient. 17. Adrenomedullin antibody or an adrenomedullin antibody nt or ADM non—lg ld according to any of the claims 1 to 16 to be used in combination with vasopressors ag. catechoiarnine. 18. Adrenomeduilin antibody or an adrenornedullin antibody fragment or ADM non—1g scaffold according to any of the claims 1 to 17 to be used in combination with intravenous fluid administration. 19. Adrenomedullin antibody or an adrenomeduflin antibody fragment or ADM non—lg scaffold ing to any of the claims 1 to 18 to be used in ation with an TNF— alpha—antibody.

. ADM antibody or an adrenomedullin antibody fragment or non—Ig—scaffold ing to any of claims 1 to 19 for use in a treatment of a patient in need thereof wherein said antibody or fragment may be administered in a dose of at least 3 pig / Kg body weight. 21. ceutical composition comprising an antibody or fragment or ld according to any of claims 1 to 20. 22. ADM antibody or an adrenomedullin antibody fragment or non—lg—scaffold according to any of claims 1 to 20 for use in a treatment of a chronic or acute disease or c condition. 23. ADM antibody or an adrenomedullin antibody fragment or non—Ig—scaffold according to claim 22 wherein said disease is sepsis.

Further embodiments within the scope ofthe present invention are set out below: Adrenomedullin ADM antibody or an adrenomedullin antibody fragment for use in therapy of a severe chronical or acute disease of a patient for the reduction of the mortality risk for said patient.

ADM dy or an adrenomedullin antibody fragment according to claim 1 wherein the antibody format is selected from the group comprising FV fragment, scFv fragment, Fab fragment, scFab fragment, (Fab)2 fragment and scFV—Fc Fusion protein.

ADM antibody or an adrenomednllin dy fragment according ciaim 1 or 2 wherein said antibody or fragment binds to the N—terminal part (aa 1—21) of adrenomedullin.

ADM antibody or an adrenomedullin antibody fragment according to any of claims 1 to 3, wherein said antibody or fragment recognizes and binds to the N—terminal end (aal) of adrenomedullin.

ADM antibody or an adrenomedullin antibody fragment according to any of claims 1 to 4, wherein said dy or fragment is an ADM stabilizing antibody or fragment that 3O enhances the tl/2 half retention time of adrenomedullin in serum, blood, plasma at least %, I3referably at least, 50 %, more preferably > 50 "/0, most Preferabl3’ > 100 %. 6. ADM antibody or an adrenomeduilin antibody fragment according to any of claims 1 to , wherein said antibody or fragment blocks the bioactivity of ADM to less than 80 %, preferably less than 50%. 7. ADM dy or an adrenomedullin antibody fragment for use in therapy of a chronical or acute disease of a t according to any of claims 1 to 6 wherein said disease is selected from the group comprising sepsis, diabetis, cancer, heart failure, shock and kidney dysfunction. 8. ADM antibody or an adrenomeduliin antibody nt for use in therapy of a chronical or acute disease of a patient ing to any of claims 1 to 7 wherein said patient is an ICU patient. 9. ADM antibody or an adrenomedullin antibody fragment for use in therapy of a chronical or acute disease of a patient according to any of claims I to 8 wherein the mortality risk is d by preventing adverse event wherein the latter are selected from the group comprising SIRS, sepsis, septic shock, organ failure, kidney e, liquid dysbalance and low blood pressure. 10. ADM antibody or an adrenomedullin antibody fragment for use in therapy of a chronical or acute disease of a patient according to any of claims 1 to 8 wherein said antibody or fragment is to be used in combination ofADM binding protein. 1]. Pharmaceutical ation comprising an dy or fragment ing to any of claims I to 10. 12. ceutical formulation according to claim 11 wherein said pharmaceutical formulation is a solution, ably a ready—to—use solution. 13. Pharmaceutical formulation according to claim 11 wherein said pharmaceutical formulation is in a freeze—dried state. 14. Pharmaceutical formulation according to any of claims 11 to 12, wherein said pharmaceutical formulation is administered intra—muscular.

Pharmaceutical formulation according to any of claims ll to 12, wherein said ceutical formulation is administered intramvascular. 16. Pharmaceutical formulation according to claim 15, wherein said pharmaceutical formulation is administered via infusion.

Further embodiments within the scope of the present invention are set out below: Adrenomedullin (ADM) antibody or an adrenomedullin dy fragment or ADM non—1g scaffold for use in therapy of a severe chronical or acute disease or acute condition of a patient for the reduction of the mortality risk for said patient wherein said antibody or nt or scaffold is a non-neutralizing ADM antibody or a non- neutralizing adrenomedullin antibody fragment or a non—neutralizing ADM non-lg scaffold.

ADM antibody or an adrenomedullin antibody fragment according to claim 1 wherein the antibody format is selected from the group comprising FV fragment, scFV fragment, Fab fragment, scFab fragment, (Fab)2 nt and scFV—Fc Fusion n.

ADM antibody or an adrenomedullin antibody fragment or an ADM non-1g scaffold according claim 1 or 2 wherein said antibody or fragment or scaffold binds to the N— terminal part (aa 1—21) of adrenomedullin.

ADM antibody or an adrenomedullin antibody fragment or an ADM non—1g ld according to any of claims 1 to 3, wherein said antibody or fragment or scaffold izes and binds to the N—terminal end (aal) of adrenomedullin.

ADM antibody or an adrenomedullin antibody fragment or an ADM non-lg scaffold according to any of claims 1 to 4, n said antibody or fragment or scaffold is an ADM stabilizing antibody or fragment or scaffold that es the half life (ti/2 half retention time) of adrenomedullin in serum, blood, plasma at least 10 %, preferably at least, 50 0/0, more preferably > 50 %, most preferably > 100 %.

ADM antibody or an medullin antibody fragment or an ADM non—lg scaffold according to any of claims 1 to 5, wherein said antibody or fragment or scaffold blocks the bioactivity ofADM to less than 80 %, preferably less than 50%.

ADM antibody or an adrenomedullin antibody fragment or an ADM non-lg scaffold for use in therapy of a cal or acute disease of a patient according to any of claims I to 6 wherein said disease is selected from the group comprising severe infections as ag. meningitis, Systemic inflammatory Response—Syndrorn (SIRS,) sepsis; other diseases as diabetis, cancer, acute and chronic vascular diseases as eg. heart failure, myocardial ll) infarction, stroke, atherosclerosis; shock as eg. septic shock and organ dysfunction as e.g. kidney dysfunction, liver ction; burnings, surgery, traumata.

ADM antibody or an adrenomedullin antibody fragment or an ADM non—lg ld for use in therapy of a chronical or acute disease of a t according to any of claims 1 to 7 wherein said e is selected from the group comprising SIRS, a severe infection, sepsis, shock ag. septic shock .

ADM antibody or an adrenomedullin antibody fragment or an ADM non-lg scaffold for use in therapy of a chronical or acute disease or acute conditiori of a patient according to any of claims 1 to 8 wherein said patient is an ICU patient. ADM antibody or an rnedullin antibody fragment or an ADM non—lg scaffold for use in therapy of a cal or acute disease or acute condition of a patient ing to any of claims 1 to 9 wherein the mortality risk is reduced by preventing an adverse event wherein the latter are selected from the group sing SIRS, sepsis, shock as tag. septic shock, acute and chronic vascular diseases as e.g. acute heart failure, myocardial infarction, stroke; organ e as e.g, kidney failure, liver failure, fluid dysbalance and low blood pressure.

. ADM antibody or an adrenomedullin antibody nt according to any of claims 1 to 9, wherein said antibody or fragment is a human monoclonal antibody or fragment that binds to ADM or an dy fragment thereof wherein the heavy chain comprises the sequences SEQ ID NO: 1 SEQ ID NO: 2 ILPGSGST SEQ ID NO: 3 TEGYEYDGFDY and wherein the light chain comprises the sequences SEQ ID NO:4 QSIVYSNGNTY SEQ ID NO: 5 SEQ ID NO: 6 FQGSHIPYT. 12. A human monoclonal antibody or fragment that binds to ADM or an antibody fragment thereof according to claim 10 wherein said antibody or nt comprises a sequence selected from the group comprising : SEQ ID NO: 7 (AM—VH-C) QVQLQQSGAELMKPGASVKISCKATGYTFSRYWIEWVKQRPGHGLEWIGEILP GSGSTNYNEKFKGKATITADTSSNTAYMQLSSLTSEDSAVYYCTEGYEYDGFD YWGQGTTLTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKR VEPKHHHHHH SEQ ID NO: 8 (AMNHl) QVQLVQSGAEVKKPGSSVKVSCKASGYTFSRYWISWVRQAPGQGLEWMGRIL PGSGSTNYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCTEGYEYDGFD YWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKR HHHH SEQ ID NO: 9 (AM—VH2—E40) QVQLVQSGAEVKKPGSSVKVSCKASGYTFSRYWIEWVRQAPGQGLEWMGRIL PGSGSTNYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCTBGYEYDGFD YWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKR VEPKHHHHHH SEQ ID NO: 10 (AM—VH3-T26—E55) SGAEVKKPGSSVKVSCKATGYTFSRYWISWVRQAPGQGLEWMGEIL PGSGSTNYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCTEGYEYDGFD YWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVT‘VPSSSLGTQTYICNVNHKPSNTKVDKR VEPKHHHHHH SEQ ID NO: 11 (AM-VH4—T26—E40—E55) QVQLVQSGAEVKKPGSSVKVSCKATGYTFSRYWIEWVRQAPGQGLEWMGEIL PGSGSTNYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCTEGYEYDGFD YWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKR VEPKHHHHHH SEQ ID NO: 12 -C) DVLLSQTPLSLPVSLGDQATISCRSSQSIVYSNGNTYLEWYLQKPGQSPKLLIYR VSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDLGVYYCFQGSHIPYTFGGGTKLE IKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNS QESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGE SEQ ID NO: 13 (AM—VLl) DVVMTQSPLSLPVTLGQPASISCRSSQSIVYSNGNTYLNWFQQRPGQSPRRLIYR VSNRDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFQGSHIPYTFGQGTKL EIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGN SQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRG SEQ ID NO: 14 (AM-VLZ—E40) 1 0 SPLSLPVTLGQPASISCRSSQSIVYSNGNTYLEWFQQRPGQSPRRLIYR VSNRDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFQGSHIPYTFGQGTKL AAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGN SQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRG 13. ADM antibody or an adrenomedullin antibody fragment or ADM non—1G scaffold for use in therapy of a chronic or acute disease of a t ing to any of the claims 1 to 12 to be used in combination with vasopressors cg. catecholamine and/ or fluids administered intravenously. 14. ADM antibody or adrenomedullin antibody fragment or ADM non—IG scaffold for use in therapy of a chronic or acute disease of a patient according to any of the claims 1 to 13 or a combination according to claim 10 to be used in combination with ADM g protein and/or further active ingredients.

. Pharmaceutical formulation comprising an dy or fragment or scaffold according to any of claims 1 to l4. l6. Pharmaceutical formulation according to claim 15 wherein said pharmaceutical formulation is a solution, preferably a ready—to-use solution. 17. Pharmaceutical formulation according to claim 15 wherein said ceutical formulation is in a freeze—dried state. 18. Pharmaceutical formulation according to any of claims 15 to 16, wherein said pharmaceutical formulation is administered muscular. 19. Pharmaceutical formulation ing to any of claims 15 to 16, wherein said pharmaceutical formulation is administered vascular. 20. Pharmaceutical formulation according to claim 19, wherein said pharmaceutical formulation is administered Via infusion. 21. ADM dy or an Adrenomedullin antibody fragment or AM non—lg scaffold, wherein said antibody or fragment or scaffold binds to the N—terminal part (aa 1—21) of Adrenomedullin in, preferably human ADM. 22. Antibody or fragment or scaffold according to claim 2, wherein said antibody or fragment or scaffold recognizes and binds to the N—terminal end (aa 1) of Adrencmedullin.

Further embodiments within the scope of the present ion are set out below: Adrenomedullin (ADM) dy or an medullin antibody fragment for use in therapy of a chronical or acute disease of a patient for prevention of organ dysfunction or organ failure.

ADM antibody or an adrenomedullin antibody fragment for use in therapy of a chronical or acute disease according to claim 1 wherein said organ is kidney.

ADM antibody or an adrenomedullin antibody fragment according to claim 1 wherein the antibody format is selected from the group comprising FV fragment, scFV fragment, Fab fragment, scFab fragment, (Fab)2 fragment and scFv-Fc Fusion protein.

WO 72510 2012/072929 4. ADM antibody or an adrenomedullin antibody nt according any of claims 1 to 3 wherein said antibody or fragment binds to the N—terminal part (aa 1—21) of mcdullin.

. ADM antibody or an adrenomedullin antibody fragment according to any of claims 1 to 4, wherein said antibody or fragment recognizes and binds to the N—terminal end (aal) of adrenomedullin. 6. ADM antibody or an adrenomedullin antibody fragment according to any of claims I to , wherein said antibody or said fragment is an ADM stabilizing antibody or fragment that es the tl/2 half retention time of adrenomedullin in serum, blood, plasma at least 10 0/0, preferably at least 50 “/0, more preferably >50 %, most preferably >100%. 7. ADM antibody or an adrenomedullin antibody fragment according to any of claims 1 to 6, wherein said antibody blocks the bioactivity of ADM to less than 80 %, preferably less than 50%. 8. ADM antibody or an adrenomedullin dy fragment for use in therapy of a chronical or acute e of a t according to any of claims 1 to 7 wherein said disease is Selected from the group comprising sepsis, diabetis, cancer, heart failure, and shock. 9. ADM antibody or an adrenomeduliin antibody fragment for use in therapy of a chronical or acute disease of a patient according to any of claims 1 to 8 wherein said patient is an ICU patient. 10. ADM antibody or an adrenomedullin antibody fragment for use in therapy of a cal or acute disease of a patient according to any of claims 1 to 9 wherein said antibody or fragment is a modulating antibody or fragment that enhances the tl/2 half retention time of adrenomedullin in serum, blood, plasma at least 10 %, preferably at least 50 %, more preferably >50 %, most preferably >100% and that blocks the bioactivity of ADM to less than 80 “/0, preferably less than 50%. 11. Pharmaceutical formulation comprising an antibody or fragment according to any of claims 1 to 10. 12. Pharmaceutical ation according to claim 11 wherein said pharmaceutical formulation is a solution, preferably a ready—to-use solution. l3. ceutical formulation according to claim 1] wherein said pharmaceutical formulation is in a freeze-dried state. 14. Pharmaceutical formulation according to any of claims 11 to 12, wherein said pharmaceutical formulation is administered intra—muscular.

. Pharmaceutical formulation according to any of claims 11 to 12, wherein said pharmaceutical ation is administered intra—vascular. 16. Pharmaceutical formulation according to claim 15, wherein said ceutical formulation is administered via infusion.

Further embodiments within the scope of the present invention are set out below: Adrenomedullin (ADM) antibody or an adrenomedullin antibody fragment or ADM non~Ig scaffold for use in therapy of a chronical or acute e or acute condition of a patient for prevention or reduction of organ dysfunction or tion of organ failure in said patient.

S‘s) ADM antibody or an medullin antibody fragment or ADM non—lg scaffold for use in therapy of a chronical or acute disease or acute disease according to claim 1 wherein said organ is kidney or liver.

ADM antibody or an adrenomedullin antibody fragment or ADM non~IG scaffold according to claim 1 or 2 wherein said ADM dy or an medullin antibody fragment or ADM non—{G scaffold is a non-neutralizing ADM antibody or a non— neutralizing adrenomedullin antibody fragment or a non-neutralizing ADM non—1G scaffold 4. ADM antibody or an adrenomedullin antibody fragment or ADM non-1C} scaffold according to any of claims 1 or 3 wherein the antibody format is selected from the group comprising FV nt, scFv nt, Fab fragment, scFab fragment, (Fab)2 fragment and scFv—Fc Fusion protein.

. ADM antibody or an adrenomedullin antibody fragment or ADM non—IG ld according any of claims 1 to 4 wherein said antibody or fragment or scaffold binds to the Nnterminal part (as. 1~21) of adrenomedullin. 6. ADM antibody or an adrenomedullin antibody fragment or ADM non-1G scaffold according to any of claims 1 to 5, wherein said antibody or fragment or scaffold recognizes and binds to the N—terminal end (aal) of medullin. ’7. ADM dy or an adrenomedullin antibody fragment or ADM non—IG ld according to any of claims 1 to 6, wherein said antibody or said fragment or scaffold is an ADM stabilizing antibody or nt or scaffold that enhances the half life (ti/2 half retention time) of adrenomedullin in serum, blood, plasma at least 10 %, preferably at least 50 %, more preferably >50 %, most preferabiy >100%. 8. ADM dy or an adrenomedullin antibody fragment or ADM non-1G scaffold according to any of claims 1 to 7, wherein said antibody or nt or scaffold blocks the bioactivity ofADM to less than 80 %, preferably less than 50%. 9. ADM antibody or an adrenomedullin antibody nt or ADM non—1G scaffold for use in therapy of a chronical or acute disease or acute condition of a patient according to any of claims 1 to 8 wherein said disease is selected from the group comprising sepsis, diabetis, cancer, heart failure, and shock.

. ADM antibody or an adrenomeduliin antibody nt according to any of ciaims 1 to 9, wherein said antibody or fragment is a human monoclonal antibody or fragment that binds to ADM or an antibody fragment thereof wherein the heavy chain comprises the sequences SEQ ID NO: 1 GYTFSRYW SEQ ID NO: 2 ILPGSGST SEQ ID NO: 3 TEGYEYDGFDY and wherein the light chain comprises the sequences SEQ ID NO:4 QSIVYSNGNTY SEQ ID NO: 5 RVS SEQ ID NO: 6 FQGSHIPYT. ll. A human monoclonal antibody or fragment that binds to ADM or an antibody fragment f according to claim 10 wherein said antibody or fragment comprises a ce selected from the group comprising : SEQ ID NO: 7 (AM—VH—C) QVQLQQSGAELMKPGASVKISCKATGYTFSRYWIEWVKQRPGHGLEWIGEILP GSGSTNYNEKFKGKATITADTSSNTAYMQLSSLTSEDSAVYYCTEGYEYDGFD YWGQGTTLTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN WO 72510 SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKR HHHH SEQ ID NO: 8 (AM—VH1) QVQLVQSGAEVKKPGSSVKVSCKASGYTFSRYWISWVRQAPGQGLEWMGRIL PGSGSTNYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCTEGYEYDGFD YWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW’N SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKR VEPKHHHHHH SEQ ID NO: 9 (AM-VH2—E40) QVQLVQSGAEVKKPGSSVKVSCKASGYTFSRYWIEWVRQAPGQGLEWMGRIL PGSGSTNYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCTEGYEYDGFD YWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKR VEPKHHHHHH SEQ ID NO: 10 (AM-VH3-T26—E55) QVQLVQSGAEVKKPGSSVKVSCKATGYTFSRYWISWVRQAPGQGLEWMGEIL PGSGSTNYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCTEGYEYDGFD YWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN GVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKR VBPKHHI—IHHH SEQ ID NO: 11 (AM—VH4nT26—E40-E55) QVQLVQSGAEVKKPGSSVKVSCKATGYTFSRYWIEWVRQAPGQGLEWMGEIL PGSGSTNYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCTEGYEYDGFD YWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKR VEPKHHHHHH SEQ ID NO: 12 (AM—VL—C) DVLLSQTPLSLPVSLGDQATISCRSSQSIVYSNGNTYLEWYLQKPGQSPKLLIYR VSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDLGVYYCFQGSHIPYTFGGGTKLE IKRTVAAPSVFIFPPSDEQLKSGTASWCLLNNFYPREAKVQWKVDNALQSGNS QESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGE SEQ ID NO: 13 (AM—VLI) DVVMTQSPLSLPVTLGQPASISCRSSQSIVYSNGNTYLNWFQQRPGQSPRRLIYR VSNRDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFQGSHIPYTFGQGTKL EIKRTVAAPSVFIFPPSDEQLKSGTASWCLLNNFYPREAKVQWKVDNALQSGN SQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRG SEQ ID NO: 14 (AM-VLZ-E40) DWMTQSPLSLP‘VTLGQPASISCRSSQSIVYSNGNTYLEWFQQRPGQSPRRLIYR VSNRDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFQGSHIPYTFGQGTKL EIKRTVAAPSVFIFPPSDEQLKSGTASWCLLNNFYPREAKVQWKVDNALQSGN SQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRG 12. ADM antibody or an adrenomedullin antibody fragment or ADM non—1G scaffold for use in therapy of a chronical or acute disease of a patient according to any of claims 1 to 11 wherein said dy or fragment or scaffold is a modulating dy or fragment or scaffold that es the half life ( t1/2 half retention time) of medullin in serum, blood, plasma at least 10 %, preferably at least 50 0/0, more preferably >50 %, most preferably >100% and that blocks the ivity of ADM to less than 80 %, preferably less than 50%. 13. ADM antibody or an adrenomedullin dy fragment or ADM non~1G scaffold for use in therapy of a chronic or acute disease or acute condition of a patient according to any of the claims 1 to 12 to be used in combination with vasopressors agcatecholamine and/ or fluids administered intravenously. 14. ADM antibody or adrenomeduliin antibody fragment or ADM non-1G scaffold for use in therapy of a chronic or acute disease or acute condition of a patient according to any of the claims 1 to 13 or a combination according to claim 13 to be used in combination with ADM binding protein and/or r active ingredients.

WO 72510 . Pharmaceutical formulation comprising an antibody or fragment according to any of claims 1 to 13. 16. Pharmaceutical formulation according to claim 14 wherein said pharmaceutical formulation is a solution, preferably a ready—to—use solution. 17. ceutical formulation according to claim 14 wherein said pharmaceutical formulation is in a freeze—dried state. 18. Pharmaceutical formulation according to any of claims 14 to 15, wherein said pharmaceutical formulation is stered rnuscular. 19. Pharmaceutical formulation according to any of claims 14 to 15, wherein said pharmaceutical formulation is administered infra-vascular.

. Pharmaceutical formulation according to claim 18, wherein said pharmaceutical formulation is administered via infusion.

EXAMPLES It should be emphasized that the antibodies, antibody fragments and non-1g scaffolds of the example portion in accordance with the invention are binding to ADM, and thus should be considered as anti—ADM antibodies/antibody fragments/non-Ig scaffolds.

Example 1 Generation of dies and determination of their affinity constants Several human and marine antibodies were produced and their affinity constants were determined (see tables I and 2).

Peptides/ conjugates for immunization: Peptides for immunization were synthesized, see Table l, (JPT Technologies, Berlin, Germany) with an additional N—terminal Cystein (if no Cystein is present within the selected ADM-sequence) e for ation of the peptides to Bovine Serum Albumin (BSA).

The peptides were ntly linked to BSA by using Sulfolink—coupling gel (Perbio—science, Bonn, Germany). The coupling procedure was performed according to the manual of Perbio.

The murine antibodies were generated according to the following method: A Baib/c mouse was immunized with lOOug e—BSA-Conjugate at day 0 and 14 (emulsified in 100111 complete ’s adjuvant) and 50ug at day 21 and 28 (in lOOul incomplete Freund’s adjuvant). Three days before the fusion experiment was performed, the animal received 50ug of the conjugate dissolved in 100 ul saline, given as one intraperitoneal and one intra—venous injection.

Spenocytes from the immunized mouse and cells of the myeioma cell line SP2/O were fused with lmi 50% polyethylene glycol for 303 at 37°C. After washing, the cells were seeded in l cell culture plates. Hybrid clones were selected by growing in HAT medium [RPMI 1640 e medium supplemented with 20% fetal calf serum and HAT-Supplement]. After two weeks the HAT medium is replaced with HT Medium for three passages ed by returning to the normal cell culture medium.

WO 72510 The cell culture supernatants were primary screened for antigen specific lgG antibodies three weeks after fusion. The positive tested microcultures were transferred into 24—well plates for propagation. After retesting, the selected cultures were cloned and recloned using the limiting~dilution technique and the isctypes were determined. (see also Lane, RD. “A short—duration polyethylene glycol fusion technique for increasing production of onal antibody-secreting hybridornas”, J. Immunol. Meth. 81: 223—228; (1985), r, B. er a1. “Glutamate oxylase (GAD) is not detectable on the surface of rat islet cells examined by cytofluorometry and complementwdependent antibody—mediated cytotoxicity of monoclonal GAD antibodies”, Horm. Metab. Res. 28: 11—15, (1996)).

Mouse monoclonal antibody production: Antibodies were produced via standard antibody production methods (Mm er a], Monoclonal Antibody Production, ATLA 25, 121, 1997,) and purified Via Protein A. The antibody purities were > 95% based on SDS gel electrophoresis analysis.

Human Antibodies Human Antibodies were produced by means ofphage display according to the following procedure: The human naive antibody gene libraries HAL7/8 were used for the isolation of recombinant single chain F—Variable domains (scFv) t adrenomedullin peptide. The antibody gene libraries were screened with a panning strategy comprising the use of peptides containing a biotin tag linked via two different spacers to the medullin peptide sequence. A mix of panning rounds using ecifically bound antigen and streptavidin bound antigen were used to minimize ound of ecific binders. The eluted phages from the third round of panning have been used for the generation of monoclonal scFv expressing Ecoli strains.

Supernatant from the cultivation of these clonal strains has been directly used for an n ELISA testing (see also Hust, M., Meyer, T., Voedisch, 13., Riilker, T., Thie, H., zal, A., Kirsch, Mi, Schiitte, M., Helmsing, S., Meier, D., Schirrmann, T., Diibel, S., 2011. A human scFv antibody generation pipeline for me research. l of Biotechnology 152, 159470; Schiitte, M., Thullier, P., Pelat, T., Wezler, X., Rosenstock, P., Hinz, D., Kirsch, M.I.,Hasenberg, M., Frank, R., Schirrmann, T., Gunzer, M., Hust, M., Diibel, 8., 2009. Identification of a putative Crf splice variant and generation of recombinant antibodies for the specific detection of Aspergillus fumigatus. PLoS One 4, e6625).

Positive clones have been selected based on positive ELISA signal for antigen and negative for streptavidin coated micro titer plates. For further characterizations the scFv open reading frame has been cloned into the expression plasmid pOPE107 (Hust et 61]., J. hn. 2011), captured from the culture atant Via immobilised metal ion affinity chromatography and purified by a size exclusion tography. y Constants To determine the affinity of the antibodies to Adrenomedullin, the kinetics of g of Adrenomedullin to immobilized antibody was determined by means of label~free surface n resonance using a Biacore 2000 system (GE Healthcare Europe GmbH, Freiburg, y). Reversible immobilization of the antibodies was performed using an antinmouse Fc antibody covalently coupled in high density to a CMS sensor surface according to the cturer's instructions (mouse antibody capture kit; GE Healthcare). (Lorenz at 4:21.,“ Functional Antibodies Targeting IsaA of Staphylococcus aureus Augment Host Immune Response and Open New Perspectives for cterial Therapy“; Antimicrob Agents Chemother. 2011 January; 55(1): 165473.) The monoclonal antibodies were raised against the below depicted ADM regions of human and murine ADM, respectively. The following table ents a selection of obtained antibodies used in further experiments. Selection was based on target region: Table 1: Wnce mtigen/lmmunogen ADM Designation Wfinity Number Region constants Kd (M) WRQSMNNFQGLRSFGCRFGTCEzi NT—H is-s x 139’ _| 'S_EQ ID: 16_iCTVQKLAHQIYQ 21—32 MR~H ”ml—2} MTV 1 WCAPRSKISPQGY~NH2 "+0—42—52 CT-H _i1.1x 10'9 SEQ ID: 18 YRQSMNQGSRSNGCRFGTC 1149 NT-M :39 iii—(rm SEQ ID: 19 CTFQKLAHQIYQ 19-31 MR-M 4.5 x 10'?U REQ n): 20 CAPRNKISPQGY—NH2 040—50 CT-M "—1 9 iii—0:9 The following is a list of further obtained onal antibodies: List of anti-ADMaantibodies Table 2: Target Source Klone numm Affinity max inhibition h] (M) ay (%) (see example 2) _| NT—M Mouse ADM/63 [TBXlO'g— Mouse ADM/364 — Mouse ADM/365 3.0111103.— Mouse 6 YMouse ADM/367 11kW1.333133 Mouse ADM/368 1.9m Mouse ADM/369 17.0 1:16:8— Mouse ADM/370 1.6 x1633" Mouse ADM/371 2.0 KW j Mouse ADM/372 2.5 1110'?“ I’— Mouse ADM/373 1.8 x10'g_ . Mouse ADM/377 1.5 x1113— % Mouse ADM/378 2.2 x102”3 Mouse ADM/379 1.6 x1073— Mouse ADM/380 [$103— Mouse ADM/381 ' Mouse ADM/382 Mouse ADM/383T L Mouse ADM/3 84 Mouse ADM/385 FMouseMouse ADM/403 ADM/395 3 Mouse ADM/396 Mouse ADM/597"“ MR-M Mouse ADM/38 muse x ADM/39 CT-M ‘ Mouse I ADM/65 CT—M Mouse ADM/66 ? NT—H Mouse WMBS ADM/34 ADM/41 ADM/42 ixDM/ZMADM/43 ADM/15 ADM/l6 ADM/17 ADM/18 Phage display ADM/A7 Phage diSplay ADM/B7 Phage display ADM/C7 Phage display ADM/G3 Phage display ADM/B6 Phage display ADM/B11 - Phage display ADM/D8 Phage display ADM/D11 Phage y ADM/G12 Generation of antibod fra entsb enz atic di estion: The generation of Fab and F(ab)2 fragments was done by enzymatic digestion of the marine full length antibody NT-M. Antibody NT~M was digested using a) the pepsin-based F(ab)2 Preparation Kit (Pierce 44988) and b) the papain—based Fab ation Kit (Pierce .

The fragmentation procedures were performed according to the instructions provided by the supplier. Digestion was carried out in case of F(ab)2—fragmentation for 8h at 370C. The Fabm ntation digestion was carried out for 1611, respectively.

Procedure for Fab Generaticn and Purification: The immobilized papain was equilibrated by washing the resin with 0.5 m1 of Digestion Buffer and centrifuging the column at 5000 X g for 1 minute. The buffer was discarded afierwards. The desalting column was prepared by removing the storage solution and washing it with digestion buffer, centrifuging it each time afterwards at 1000 x g for 2 minutes. 0.51111 of the prepared IgG sample where added to the spin column tube ning the equilibrated Immobilized Papain. tion time of the digestion reaction was done for 16h on a tabletop rocker at 37°C. The column was centrifuged at 5000 X g for 1 minute to separate digest from the Immobilized Papain. Afterwards the resin was washed with 0.51111 PBS and centrifuged at 5000 x g for 1 minute. The wash fraction was added to the digested antibody that the total sample volume was 1.0ml. The NAb Protein A Column was equilibrated with PBS and IgG Elation Buffer at room temperature. The column was fuged for 1 minute to remove storage solution (contains 0.02% sodium azide) and equilibrated by adding 2mi of PBS, centrifuge again for 1 minute and the flow-through discarded. The sample was applied to the column and resuspended by inversion. Incubation was done at room temperature with end~ over—end mixing for 10 s. The column was centrifuged for 1 minute, saving the flow- through with the Fab fragments.

(References: r, A. and Harris, R. (1983). J. lmrnunol. Meth. 59, 199—203.; Lindner I. et a1. (2010) {alpha}2—Macroglobulin inhibits the malignant properties of astrocytoma cells by impeding {beta}~catenin signaling. Cancer Res. 70, 277—87.; Kaufmann B. et at. (2010) Neutralization of West Nile virus by cross—linking of its surface proteins with Fab fragments of the human monoclonai antibody CR4354. PNAS. 107, 18950—5; Chen X. et a1. (2010) Requirement of open ece conformation for activation of leukocyte integrin axBZ.

PNAS. 107, 14727—32; Uysal H. et a1. (2009) Structure and pathogenicity of dies specific for citrullinated collagen type 11 in experimental arthitis. J. Exp. Med. 206, 449—62.; Thomas G. M. et al. (2009) Cancer erived microparticles bearing P—selectin glycoprotein ligand 1 accelerate thrombus formation in vivo. J. Exp. Med. 206, 1913—27.; Kong F. et a1. (2009) Demonstration of catch bonds between an integrin and its ligand. 1. Cell Biol. 185, 1275—84.) Procedure for tion and purification of F1 ab’]; nts: The lized Pepsin was equilibrated by washing the resin with 0.5 m1 of Digestion Buffer and centrifuging the column at 5000 X g for 1 minute. The buffer was discarded afterwards. The desalting column was prepared by ng the storage solution and washing it with digestion buffer, fuging it each time afterwards at 1000 x g for 2 minutes. 051111 of the prepared IgG sample where added to the spin column tube containing the equilibrated Immobilized . Incubation time of the digestion reaction was done for 16h on a tabletop rocker at 37°C. The column was fuged at 5000 X g for 1 minute to separate digest from the lized Papain. Afterwards the resin was washed with 0.5mL PBS and centrifuged at 5000 X g for 1 minute. The wash fraction was added to the digested dy that the total sample volume was 1.01111. The NAb Protein A Column was equilibrated with PBS and igG Elation Buffer at room temperature. The column was centrifuged for 1 minute to remove storage solution (contains 0.02% sodium azide) and equilibrated by adding ZinL of PBS, centrifuge again for 1 minute and the rough discarded. The sample was applied to the column and resuspended by inversion. Incubation was done at room temperature with end- over—end mixing for 10 minutes. The column was centrifuged for 1 minute, saving the flow— h with the Fab fragments.

(References: Mariam, M., at al. . A new enzymatic method to obtain high-yield F(ab')2 suitable for clinical use from mouse lgGl. Mollmmunol. 28: 69—77.;Beale, D. (1987).

Molecular ntation: Some applications in immunology. Exp Comp Immunol 11:287— 96.; on, J.R., et a]. (1972). A fragment ponding to the CH2 region of immunoglobulin G (IgG) with complement fixing activity. FEBS Letters 24(3):318~22.; Kerbel, RS. and Elliot, BE. (1983). Detection of Fc receptors. Meth Enzymol 93:113—147.; Kulkarni, P.N., er al. (1985). ation of methotrexate to IgG antibodies and their F(ab')2 fragments and the effect of conjugated methotrexate on tumor growth in vivo. Cancer Immunol Immunotherapy 1922114.; Lamoyi, E. (1986). Preparation of F(ab’)2 Fragments from mouse IgG of various subclasses. Meth Enzymol 121:652—663.; Parham, P., et at. (1982). Monoclonal antibodies: purification, fragmentation and application to structural and functional s of class I MHC antigens. J Immunol Meth 53:133—73; Raychaudhuri, (3., er al. (1985). Human lgGi and its Fc fragment bind with different affinities to the Fc receptors on the human U937, HL-60 and ML—l cell lines. Mol Immunol 22(9):1009—19.; Rousseaux, 1., er a1. (1980). The differential enzyme sensitivity of rat immunoglobulin G subclasses to papain an . Moi Immunol 171469—82; Rousseaux, J., at a]. (1983). Optimal condition for the preparation of Fab and F(ab’)2 fragments from monoclonal IgG of different rat IgG subclasses. J Immunol Meth 64:141—6.; Wilson, K.M., er al. (1991). Rapid whole blood assay for HIV—1 sitivity using an Fab~peptide conjugate. J Immunol Meth 138:111—9.) NT~H~Antibody Fragment Humanization The antibody fragment was humanized by the CDR—gratting method (Jones, P. T., Dear, P. 1-1., Foote, 3., Neuberger, M. 8., and Winter, G. (1986) Replacing the complementarity— determining regions in a human antibody with those from a mouse. Nature 321, 522—525).

The following steps where done to achieve the humanized sguence: Total RNA extraction: Total RNA was extracted from NT—I—I hybridomas using the Qiagen kit.

First-round RT—PCR: QIAGEN® OneStep RT—PCR Kit (Cat No. 210210) was used. RT~PCR was performed with primer sets specific for the heavy and light . For each RNA sample, 12 dual heavy chain and 11 light chain RT—PCR reactions were set up using degenerate forward. primer mixtures covering the leader sequences of variable regions. e primers are located in the constant regions of heavy and light chains. No restriction sites were engineered into the primers. on Setup: 5x QIAGEN® OneStep RT—PCR Buffer 50 pt], dNTP Mix ining 10 mM of each dNTP) 0.8 ul, Primer set 05 n1, ® OneStep RT—PCR Enzyme Mix 0.8 til, Template RNA 2.0 al, RNase—free water to 20.0 Ml, Total volume 20.0 id PCR condition: Reverse transcription: 50°C, 30 min; Initial PCR activation: 95°C, 15 min g: 20 cycles of 94°C, 25 sec; 54°C, 30 sec; 72°C, 30 sec; Final extension: 72°C, 10 min Second—round semi-nested PCR: The RTmI’CR products from the first—round reactions were further amplified in the ~round PCR. 12 individual heavy chain and 11 light chain RT- PCR reactions were set up using semi—nested primer sets specific for antibody variable regions. on Setup: 2}: PCR mix 10 ill; Primer set 2 pl; First—round PCR product 8 pl; Total volume 20 u]; Hybridoma Antibody Cloning Report PCR condition: Initial denaturing of 5 min at 95°C; 25 cycles of 95°C for 25 sec, 57°C for 30 sec, 68°C for 30 sec; Final extension is 10 min 68°C.

After PCR is finished, run PCR reaction samples onto agarose gel to visualize DNA fragments ed.After sequencing more than 15 cloned DNA nts amplified by nested RT-PCR, several mouse antibody heavy and light chains have been cloned and appear correct. Protein sequence alignment and CDR analysis identifies one heavy chain and one light chain. After alignment with homologous human framework sequences the ing humanized sequence for the variable heavy chain is the following: see figure 6 (As the amino acids on positions 26, 40 and 55 in the variable heavy chain and amino acid on position 40 in the variable light are critical to the binding ties, they may be reverted to the murine original. The resulting candidates are depicted below) (Padlan, E. A. (1991) A possible procedure for reducing the immunogenicity of antibody variable s while preserving their ligand—binding properties. Mol. Immunol. 28, 489—498.; Harris, L. and Bajorath, l. (1995) Profiles for the analysis of immunoglobulin sequences: Comparison of V gene ups. Protein Sci. 4, 306810.).

Annotation for the dy fragment ces (SEQ ID NO: 744): bold and underline are the CDR l, 2, 3 in chronologically arranged; italic are constant regions; hinge regions are highlighted with bold letters and the histidine tag with bold and italic letters; framework point mutation have a grey letter—background.

SEQ ID NO: 7 (AM—VI—LC) QVQLQQSGAELMKPGASVKISCKATGYTFSRYWIEWVKQRPGHGLEWIGEILPGSG §1NYNEKFKGKATITADTSSNTAYMQLSSLTSEDSA GYEYDGFDYWGQGTTLT VSSASTKGPSVFPLAPSSKSTSGGTAALGCL VKDYFPEPVTVSWNSGALTSGVHTFPA VLQS SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKHHHHHH SEQ ID NO: 8 (AM—VH1) QVQLVQSGAEVKKPGSSVKVSCKASGYTFSRYWISWVRQAPGQGLEWMGRILPGS QflNYAQKFQGRVTITADESTSTAYMELSSLRSEDTA GYEYDGFDYWGQGTTV TVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQ SSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKHHHHHH SEQ ID NO: 9 (AM—VH2~E40) QVQLVQsGAEVKKPGSSVKVSCKAsGYTFSRYWIfiWVRQAPGQGLEWMGRILPGS QINYAQKFQGRVTITADESTSTAYMELSSLRSEDTA WYCTEGYEYDGFDYWGQGTTV TVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQ SSGLYSLSSVVTVPSSSLGTQTYICNWVHKPSNTKVDKRVEPKHHHHHH SEQ ID NO: 10 (AM-VH3—T26—E55) QVQLVQSGAEVKKPGSSVKVSCKAEEGYTFSRYWISWVRQAPGQGLEWMGWETILPGS FQGRVTITADESTSTAYMELSSLRSEDTA VYYCTEGYEYDGFDYWGQGTTV TVSSASTKGPSVFPLAPSSKSTSGGTAALGCL VKDYFPEPVTVSWNSGALTSGVHTFPA VLQ SSGLYSLSSVVTVPSSSLGTQWICNVZ’VHKPSNTKVDKRVEPKHHHHHH SEQ ID NO: 11 (AM—VH4~T26—B40—E55) QVQLVQSGAEVKKPGSSVKVSCKASEEGYTFSRYWI:WVRQAPGQGLEWM EEILPGS QSENYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCTEGYEYDGFDYWGQGTTV TVSSASTKGPSVFPLAPSSKSTSGGTAALGCL VKDYFPEPVTVSWWSGALTSGVHTFPA VLQ LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKHHHHHH SEQ ID NO: 12 (AM—VL—C) DVLLSQTPLSLPVSLGDQATISCRSSQSIVYSNGNTYLEWYLQKPGQSPKLLIYMN RFSGVPDRFSGSGSGTDFTLKISRVEAEDLGVYYCFQGSHIPYTFGGGTKLEIKRTVA APSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDS KDS'I‘YSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 13 I) DVVMTQSPLSLPVTLGQPASISCRSSQSIVYSNGNTYLNWFQQRPGQSPRRLIYEXfiN RDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFQGSHIPYTFGQGTKLEIKRTVA APSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDS KDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 2O SEQ ID NO: 14 (AM—VLZ-FAO) DWMTQSPLSLPVTLGQPASISCRSSQSIVYSNGNTYLEZZWFQQRPGQSPRRLIYELSN DRFSGSGSGTDFTLKISRVEAEDVGVYYCFQGSHIPYTFGQGTKLEIKRTVA APSVFIFPPSDEQLKSGTASWCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDS KDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC Example 2 Effect of selected anti~ADM-antibodies on anti-ADM-bioactivity The effect of selected ADM-antibodies on ADM—bioactivity was tested in an human recombinant Adrenomedullin receptor CAMP functional assay (Adrenomedullin Bioassay).

Testing of antibodies targeting human or mouse adrenomedullin in human recombinant Adrenomedullin receptor CAMP functional assay (Adrenomedullin Bioassay) Materials: Cell line: CHO—Kl Receptor: rnedullin (CRLR + RAMPB) Receptor Accession Number Cell line: CRLR: U17473; RAMP3: AJ001016 CHO—Kl cells expressing human recombinant adrenomedullin or (FAST—027C) grown prior to the test in media Without antibiotic were detached by gentle flushing with PBS—EDTA (5 mM EDTA), recovered by centrifugation and resuspended in assay buffer (KRH: 5 1nM KCl, 1.25 mM MgSO4, 124 mM NaCl, 25 mM HEPES, 13.3 mM Glucose, 1.25 mM KH2P04, 1.45 mM CaClB, 0.5 g/l BSA).

Dose response curves were med in parallel with the reference agonists (hADM or mADM).

Antagonist test (96well): For antagonist testing, 6 n] of the reference agonist (human M) or rnouse (0,67nM) adrenomedullin) was mixed with 6 pl of the test samples at different antagonist dilutions; or with 6 til buffer. After incubation for 60 min at room temperature, 12 at of cells (2,500 cells/well) were added. The plates were incubated for 30 min at room temperature. After addition of the lysis buffer, percentage of DeltaF will be estimated, according to the manufacturer specification, with the HTRF kit from Cis—Bio International (cat n°62AM2 PEB). hADM 22—52 was used as reference antagonist. dies testing TRF assay The anti—hnADM antibodies (NT-H, MR-H, CT—I—I) were tested for antagonist activity in human recombinant adrenomedullin receptor 027C) CAMP functional assay in the presence of 5.63nM Human ADM 1—52, at the following final antibody concentrations: lOOug/inl, 20ug/ml, 4ug/nil, 0.8itg/ml, 0.16ng/mi.

The anti—m—ADM antibodies (NT—M, MR—M, CT—M) were tested for antagonist activity in human recombinant adrenorneduliin receptor (FAST-027C) CAMP onal assay in the presence of 0.67nM Mouse ADM 1-50, at the following final antibody trations: lOOug/ml, 20ug/ml, 4ug/nil, 0.8ug/ml, 0.16ng/ml. Data were d relative inhibition vs. antagonist concentration (see figs. 3a to 31). The maximal inhibition by the individual antibody is given in table 3.

Table 3: Antibody Maximal tion ofADM bioactivity (ADM-Bioassay) (0/0) NT—H 38 MR~H 73 j CT—H 100 NT—M FAB 26 NT-M FABZ 28 NT-M 45 MR~M 66 CIT—M 100 Non specific mouse lgG 0 §__l Data for stabilization of hADM by the anti-ADM antibody The stabilizing effect of human ADM by human ADM antibodies was tested using a hADM immunoassay.

Immunoassay for the quantification of human Adrenomedullin The technology used was a sandwich coated tube luminescence immunoassay, based on nium ester labelling.

Labelled compound (tracer): 100ng (lOOul) CT—H (lmg/ ml in PBS, pH 7.4? AdrenoMed AGGerniany) was mixed with lOul Acridiniurn NHSmester (lmg/ ml in acetonitrile, InVent GmbH, Germany) (EP 0353971) and incubated for 20min at room temperature. Labelled CT— H was purified by Gelufiltration HPLC on Bio—Sil® SEC 400-5 (Bio—Rad Laboratories, Inc, USA) The d CT—H was diluted in (300 mmol/L potassiumphosphate, 100 mmol/L NaCl, 10 mmol/L NanEDTA, 5 g/L Bovine Serum Albumin, pH 7.0). The final tration was approx. 0 relative light units (RLU) of labelled compound (approx. 20mg labeled WO 72510 antibody) per 200 uL. Acridiniumester chemiluminescence was measured by using an AutoLumat LB 953 (Berthold Technologies GmbH & Co. KG).

Solid phase: Polystyrene tubes (Greiner Bio—One International AG, a) were coated (18h at room temperature) with MR—H (AdrenoMed AG, Germany) (1.5 ug MR—H/O.3 mL 100 mmol/L NaCl, 50 mmol/L TRIS/HCI, pH 7.8). After blocking with 5% bovine serum ne, the tubes were washed with PBS, pH 7.4 and vacuum dried.

Calibration: The assay was ated, using dilutions ofhADM (BACHEM AG, Switzerland) in 250 mmol/L NaCl, 2 g/L Triton X-100, 50 g/L Bovine Serum Albumin, 20 tabs/L Protease Inhibitor Cocktail (Roche Diagnostics AG, Switzerland» llADM assay: 50 ul of sample (or calibrator) was pipetted into coated tubes, after adding labeleld CT—H (200ul), the tubes were incubated for 4h at 40C. Unbound tracer was removed by washing 5 times (each 1ml) with washing solution (20mM PBS, pH 7.4, 0.1 % Triton X—l 00).

Tube—bound chemiluminescence was measured by using the LB 953 Figure 4 shows a typical hADM dose/ signal curve. And an hADM dose signal curve in the presence of 100 ug/mL antibody NT~H.

NT—H did not affect the described hADM immunoassay.

Stability of human Adrenomedullin: Human ADM was diluted in human e plasma (final concentration lOnM) and ted at 24 0C. At selected time points, the degradation of hADM was stopped by freezing at —20 0C. The incubation was performed in absence and presence of NT-H (lOOug/rnl) . The remaining hADM was quantified by using the hADM immunoassay described above.

Figure 5 shows the stability of hADM in human plasma (citrate) in absence and in the presence of NT—H antibody. The half life ofhADM alone was 7,8h and in the presence ofNT— H, the half life was 18,311. (2.3 times higher ity).

Example 4 Sepsis Mortalitljearly treatment; Animal model 12—15 week old male C57Bl/6 mice (Charles River Laboratories, y) were used for the study. Peritonitis had been surgically d under light isofluran anesthesia. lncisions were made into the left upper quadrant of the peritoneal cavity (normal location of the cecum). The cecum was exposed and a tight ligature was placed around the cecum with sutures distal to the insertion of the small bowel. One re wound was made with a 24—gauge needle into the cecum and small amounts of cecal contents were expressed through the wound. The cecum was replaced into the peritoneal cavity and the laparotomy site was closed. Finally, animals were returned to their cages with free access to food and water. 500p] saline were given so. as fluid repiacernent.

Application and dosage of the compound (NT—M, MR—M, CT-M) Mice were treated immediately afier CLP (early treatment). CLP is the abbreviation for cecal ligation and puncture (CLP).

Study groups Three compounds were tested versus: vehicle and versus control compound treatment. Each group contained 5 mice for blood g after 1 day for BUN (serum blood urea nitrogen test) determination. Ten further mice per each group were followed over a period of 4 days.

Group Treatment (lOul/ g bodyweight) dose/ ap: 1 NT—M, 0.2 mg/mi survival over 4 days 2 MR-M, 0.2 mg/ml survival over 4 days 3 CTuM, 0.2 rug/ml survival over 4 days 4 nonuspecific mouse lgG, 0.2 nag/ml survival over 4 days control — PBS 10ul/g bodyweight survival over 4 days Clinical chemistry Blood urea nitrogen (BUN) concentrations for renal function were measured baseline and day l alter CLP. Blood samples were obtained from the cavernous sinus with a capillary under light ether anaesthesia. Measurements were performed by using an AU 400 Olympus nalyser. The 4—day mortality is given in table 4. The average BUN concentrations are given in table 5.

Table 4: W4 day mortality survival (%) as l lET—Mnon-specific mouse IgG 0 MR—M 30 NT—M 70 Table 5: Average from 5 animals BUN pre CLP (mM) BUN day 1 (mM) PBS 8.0 i 23.2 .l_n ecific mouse IgG 7.9 i73.515.5 CT-M 7.8 MR-M 8.1 We NT—M 8.8 it can be seen from Table 4 that the NT—M antibody d mortality erably. After 4 days 70 9/0 of the mice survived when treated with NT—M antibody. When treated with MR—M antibody 30 ”/0 of the animals survived and when treated with CT—M antibody 10 “/0 of the animals survived after 4 days. in contrast o all mice were dead after 4 days when treated with unspecific mouse IgG. The same result was obtained in the control group where PBS (phOSphate buffered saline) was administered to mice.

The blood urea nitrogen or BUN test is used to te kidney function, to help diagnose kidney disease, and to monitor patients with acute or chronic kidney dysfunction or failure.

The results of the S—BUNr Test ed that the NT—M dy was the most effective to protect the kidney.

Sepsis Mortality (late treatment) Animal model 12—15 week old male C57Blf6 mice es River Laboratories, Germany) were used for the study. Peritonitis had been surgically induced under light isofluran anesthesia. Incisions were made into the left upper quadrant of the peritoneal cavity (normal location of the cecum). The cecum was exposed and a tight ligature was placed around the cecum with sutures distal to the insertion of the small bowel. One puncture wound was made with a ge needle into the cecum and small amounts of cecal contents were expressed through the wound. The cecum was replaced into the peritoneal cavity and the laparotorny site was closed. Finally, animals were returned to their cages with free access to food and water. SOOul saline were given so. as fluid replacement. 2O Application and dosage of the compound (NT-M FAB2) NT-M FABZ was tested versus: e and versus control compound treatment. Treatment was performed after full development of sepsis, 6 hours after CLP (late treatment). Each group contained 4 mice and were followed over a period of 4 days.

Group Treatment (lOul/ g bodyweight) dose/ -Up: Study groups 1 NT—M, FABZ 0.2 mg/ml survival over 4 days 2 control : non—specific mouse IgG, 0.2 mg/ml survival over 4 days 3 vehicle: » PBS lOul/g bodyweight survival over 4 days WO 72510 Table 6: 4Pd: mortality survival (%) Non—specific mouse IgG NT-M FABZ It can be seen from Table 6 that the NT—M FAB 2 antibody d mortality considerably.

After 4 days 75 % of the mice survived when treated with NTuM FAB 2 antibody. In contrast thereto all mice were dead after 4 days when treated with non—specific mouse IgG. The same result was obtained in the control group Where PBS (phosphate buffered saline) was administered to mice.

Example 5 Incremental effect of DM antibody in CLP—animals on top of antibiotic treatment and circulation stabilization via catecholamines as well as regulation of fluid balance.

Animal model In this study male 6 mice (8—12 weeks, ) were utilized. A polymicrohial sepsis d by cecal ligation and puncture (CLP) was used as the model for studying septic shock ((Albuszies G, er al: Effect of increased cardiac output on hepatic and intestinal microcirculatory blood flow, oxygenation, and metabolism in hyperdynamic murine septic shock. Crit Care Med 2005;33:2332-8), (Albuszies G, at al: The effect of iNOS deletion on hepatic gluconeogenesis in hyperdynamic murine septic shock. Intensive Care Med 2007;33:1094—101), (Barth E, et al: Role of iNOS in the reduced responsiveness of the myocardium to catecholainines in a hyperdynamie, murine model of septic shock. Crit Care Med 2006;34:307—13), (Baumgart K, at al: Effect of SOD—1 over—expression on myocardial function during resuscitated murine septic shock. Intensive Care Med 2009;35:344-9), (Baumgart K, er of: Cardiac and metabolic effects of hypothermia and inhaled H28 in anesthetized and ventilated mice. Crit Care Med 2010;38:588—95), (Simkova V, et al: The effect of SOD—1 xpression on hepatic gluconeogenesis and whole—body glucose oxidation during resuscitated, norrnotensive murine septic shock. Shock 2008;30:578-84), r F, at al.: Inflammatory effects of ermia and inhaled H28 during resuscitated, WO 72510 hyperdynamic mun'ne septic shock. Shock, im Druck), (Wagner F, er al: Effects of intravenous H28 after murine blunt chest trauma: a prospective, randomized controlled trial.

Crit Care 2011, submittes for publication)).

After weighing, mice were anesthetized by intraperitoneal injection of 120 ug/g Ketarnin, 1.25 ug/g Midazolam and 0.25 pig/g Fentanyl. During the surgical ure, body ature was kept at C. A lcm midline abdominal section was performed to get access to the cecum. The cecum then was ligated with 3—0 silk tie close to the basis and a single puncture with a 18-gauge needle was applied. The cecum was returned and the incision was closed again (4-0 tie). For the compensation of perioperative loss of liquids, 0.5 ml lacted Ringer’s solution with lug!g Buprenorphin as analgetic was injected subcutaneously in dorsal dermis. For antibiosis the mice received Ceftn'axon 30ug/g and Clindamycin 30ug/g aneously via the lower extremities.

After CLP surgery the animal were kept in an adequately heated environment with water and food ad iibitum.

The covering of liquid requirements were ensured by a dorsal subcutaneous injections with 0.5 ml lactated ’s solution with 4 ug/g glucose and orphin lug/g, which were applied in an 8 hour cycle, after short term anesthesia by isofluran. In addition, antibiosis was maintained by subcutaneous injections of Ceftriaxon 30ng/g and Clindamycin 30ug/g via the lower extremities.

Dosing of test substances Early treatment Immediately after the CLP surgery and closing of the incision, the test substance antibody NT—M was applied in a concentration of 500 ug/ml in ate buffered saline (PBS) Via injection into the penis vein for a dose of 2mg per kg body weight (dose volume 88—120 til) (5 animals).

Late treatment After full Sepsis development, 15.5h after CLP surgery, animals were etized as described above and NT—M was applied in a concentration of 500 ug/ml in phosphate buffered saline (PBS) via injection into the penis vein for a dose of 2mg per kg body weight (dose volume 88—120 ul) (3 animals).

The control group (6 animals) received a corresponding amount of the vehicle PBS on without dy (4ul/g, 88420 ul) immediately after CLP surgery.

Study groups and experimental setting Murine septic shock model under intensive care monitoring: Three groups with 3, 5 and 6 animals were monitored. Group 1 (5 animals) received the antibody NT—M 15.5h alter CLP, group 2 received the antibody NT—M immediately after CLP surgery and group 3 received a comparable amount of PBS (4ul/g). 16 hour incubation post CLP (to allow the polymicrobial sepsis to progress), the experiment was continued with monitoring and interventions comparable to an intensive medical care regime. Therefore, after ng the s were anesthetized as described in the CLP surgery part (except the late treated animals, which were anesthized before treatment). Body temperature was maintained at 37-38°C for the rest of the experiment. After a otomy and intubation, respiration was monitored and supported by laboratory animal lung ventilator Flexivent®, (Emka Technologies, FiO2 0,5, PEEP 10 H20, VT Sui/g, l:E 121,5, AF 70-140 depending on temperature). esia was ined throughout the experiment via the ated vena jugularis externa dextra with a continuous infusion of Ketamin 30 ug/gxh and Fentanyl 0.3 ug/gxh.

Furthermore, the right aorta carotis communis was cannulated for continuous monitoring of heart rate and the mean arterial pressure (MAP). The mean arterial re was maintained at MAP > 65 mmHg via intravenous (V. jugularis) infusion of colloids (80 uL/gxh, Hextend®) and, if needed, Noradrenaiin dissolved in colloids as vasopressor. Blood samples (120 pl) were taken via the cannulated A. carotis at 0 and 4 hours for determination of creatinine. The bladder was punctured and urine was ted via a r catheter. The experiment was either terminated after 6 hours or prior to this, if the MAP > 65 mmHg (V. jugularis) could not be maintained with the vasorpressor dosing.

Measured parameters The following parameters were ed and analyzed: Total consumption of noradrenalin (pg NA/g), consumption rate of noradrenalin (ug NA/g/h), total volume of urine collected during the experiment, creatinine tration ) at the end of the experiment and mean creatinine clearance (uL/min).

Table 7: Total consumption of consumption rate of Noradrenalin (ug NA]g) Noradrenalin (ug NA/g/h) Control (mouse lgG) (N26) 0.17 Myg 0.032 ug/hfg NT—M (N=5) early treatment 0.07 ug/g 0.012ug/h/g Relative change (early treatment, 59% 62.5% amelioration) (59%) (62.5%) NT-M (N33) late treatment 0.04 ug/g 0.0075 ug/h/g Relative change (late treatment, 76,5% 76,5% amelioration) (76.5%) (76.5%) The olamine requirement was ed after administration of either non specific mouse lgG to a total of 6 mice as control group, NT—murine antibody to a group of 5 mice ately after CLP (early treatment) or NT-murine antibody to a group of 3 mice 15.5h after CLP (late treatment).

The reduction of the catecholamine requirement is a measure for the stabilization of the circulation. Thus, the data Show that the ADM antibody, especially the NT—M antibody, leads to a considerable stabilization of the circulation and to a considerable reduction of the olamine requirement. The circulation—stabilizing effect was given in early treatment (immediately after CLP) and treatment after full sepsis development (late ent) (see fig.

Regulation of Fluid Balance More positive fluid balance both early in resuscitation and cumulatively over 4 days is associated with an increased risk of mortality in septic shock. The control of the liquid balance is of utmost importance for the course of disease of patients having sepsis. (s. Boyd er al, 2011). lling the liquid balance of critical ill patients remains as a substantial challenge in intensive care medicine. As can be seen in table 8 treatment of mice after CLP (experimental ures see “Animal Model”) with NT—M antibody lead to an enhancement of the total volume of urine excreted. The urine secreted was approx. three times higher in NTHMJcreated animals compared to nonutreated mice. The positive treatment effect was given in early— and in late treatment. The fluid balance was improved by about , also in both, early and late treatment. Thus, the data show that the use ofADM antibody, especially the use ofNT ADM antibody, is favorable for regulating the fluid balance in patients. (see table 8 and figures 8 and 9).

Table 8 Urine average Fluid balance average volume/ g body / gbody weight weight Control (mouse igG) 0.042 nil/g 0,23 nil/g NTFM early (N=5) 0,18 mlfg Relative change early + 186% —21.7%% treatment NT—M late (N:3) 0,16 ml/g Relative change late + 198% treat111CDT Improvement of kidneyfunction The combination of acute renal failure and sepsis is associated with a 70 percent mortality, as compared with a 45 percent mortality among patients with acute renal failure alone. (Schrier and Wang, “Mechanisms of Disease Acute Renal Failure and Sepsis”; The New England Journal of ne; 351 :159—69; 2004). Creatinine concentration and creatinine clearance are standard laboratory parameters for ring kidney (dys)function (Jacob, “Acute Renal e”, Indian J. Anaesth; 47 (5): 367-372; 2003). Creatinine and creatinine clearance data from above described animal experiment (early treatment) are given in Table 9.

Table 9 Kidney function: creatinine mean creatinine tration clearance (uL/min) (Mg/mL) control mouse IgG (MW) 2.6 rag/ml 174 til/min NT—M (MW) 1.5 rig/ml 373 til/min Relative change -42% +1 14% (amelioration) (42%) (114%) In comparision to control septic animals, the creatinine concentration was lowered by 42% and the creatinine clearance was improved by more than 100% as a result ofNT—M treatment (Table 9). The data show that the stration of ADM—antibody, ally NT—M, leads to an improvement of kidney on.

Improvement of liver inflammatory status Liver tissue for control and early treated s was homogenized and lysed in lysing buffer.

For cell t preparation, cells were resuspended, lysed on ice, and centrifuged. The supernatant (protein extract) was stored at -80 0C. Activation of r factor kappa-light~ chain gene enhancer in B cells (NF-KB) was ined as previously described using an electrophoretic mobility shift assay (EMSA)1,2. Cell extracts (mug) were incubated on ice with poly—doxy—inosinic—deoxyrcytidylic acid (poly—deC) and 32P-labeied double stranded oligonucleotide (Biomers, Ulm, Germany) containing the NF—KB (HIV KBsite) (5’- GGATCCTCAACAGAGGGGACTTTCCGAGGCCA—3’). Complexes were separated in native polyacrylamide gels, dried and exposed to X—ray films. A phosphorimager and image analyzer software (AIDA Image Analyzer; Raytest) was used to quantify the ctively labeled NF—KB by densitometry. For comparison between individual gels, the intensity of each band was related to that of simultaneously loaded control animals which had not undergone 2012/072929 surgical instrumentation and CLP. Therefore, the EMSA data are expressed as fold increase over l values. Statistics: All data are presented as median (range) unless otherwise stated differences between the two groups were analyzed with the Mann—Whitney rank sum test for unpaired samples. Results: The animals treated with NT—M ted with significantly attenuated liver tissue NF-KB activation (2.27 (1.97—2.53)) compared to vehicle animals (2.92 (2.50—3.81)) (p<0.001) (see figure 10).

References: l. Wagner F, Wagner K, Weber S, Stahl B, l MW, Huber-Lang M, Seitz DH, Asfar P, Calzia E, Senftleben U, Gebhard F, Georgieff M, acher P, Hysa V: Inflammatory effects ofhypothermia and inhaled H28 during resuscitated, hyperdynamic murine septic shock. Shock 201 l ;3 5(4) :396—402 22. Wagner F, rle A, Weber S, Stahl B, McCook O, Knoferl MW, Huber-Lang M, Seitz DH, Thomas J, Asfar P, Szabo C, Molier P, Gebhard F, Georgieff M, Calzia E, Radermacher P, Wagner K: Cardiopulmonary, histologic, and inflammatory effects of intravenous Na2S afier blunt chest traumauinduced lung contusion in mice. J Trauma 2011;71(6):1659-67 grannies In vivo side effect determination of dy NT-M 12—15 week old mate C57Bl/6 mice (Charles River Laboratories, Germany) were used for the study. 6 mice were treated with (10ul/ g bodyweight) dose of NT—M, 0.2 mg/ml. As l, 6 mice were treated with (Mini/g body ) PBS. Survival and physical condition was monitored for 14 days. The mortality was 0 in both groups, there were no differences in physical condition between NT-M and control group.

Example 7 Gentamicin-induced nephrotoxicigg A nonwseptic acute kidney injury model has been established, which makes use of the nephrotoxicity induced by Gentamicin (Chiu P] S. Models used to assess renal functions. Drug 2012/072929 p Res 32:247-255, 1994.). This model was used to assess Whether treatment with anti— Adrenomedullin antibody can improve kidney function.

The experiment was performed as follows: Effect of a NT-M on Gentamicinulnduced Nephrotoxicity in Rats Study Design: Gentarnicina + NT-M aGentamicin at 120 mg/kg intramuscularly for 7 days (days 0-6). bVehicle; injected intravenously (i.v.) 5 min before gentamicin on Day 0, followed by injections on Days 2, 4, and 6. cNT—M at 4 mg/kg was ed intravenously (i.v.) 5 min before nicin on Day 0, followed by 2 nag/kg i.v. on Days 2, 4, and 6. dPlasma samples were collected in EDTA tubes (Days 1 and 3 before Test and Control e: 100 pl; Day 7:120 n1. 24h urine collection on ice is ted afier gentamicin on Day 0, followed by Days 2 and 6; blood collection on days 1, 3, and 7.

Groups of 8 male Sprague—Dawley rats weighing 250 i 20 g were employed. Animals were challenged with gentarnicin at 120 mg/kg i.m. for seven consecutive days (Groups 1 and 2).

Test compound (anti-adrenomedullin antibody NT~M) and vehicle (phosphate buffered saline) were injected intravenously 5 min before gentamicin on day 0, followed by injection on days 2, 4, and 6. Body weights and clinical signs were monitored daily. Twenty—four (24) hour urine collections on ice were performed on Days 0, 2, and 6. Urine specimens were assayed for concentrations of Na+ and Kr, and creatinine. Blood s for clinical chemistry were collected on Days 1 (before gentamicin), 3 (before gentamicin), and 7. Serum electrolytes (Na+ and K+), creatinine, and BUN were the primary analytes that were monitored for assessing renal function. Plasma samples were collected in EDTA tubes (Days l and 3:100 til; Day 7:120 n1). Creatinine clearance was calculated. Urine volume, urinary electrolytes, and creatinine are sed as amount excreted per 100 g of animal body weight. All animals were sacrificed on Day 7. Kidneys were d.

Urine collection. The animals were placed in individual cages where urine was collected for 24 h on Day 0, Day 2, and Day 6. Urine volume, urinary Na+, K+, and creatinine were measured.

Endogenous creatinine clearance was calculated as follows: CCr (ml/24 h) = [UCr ) x V (ml/24 h)] / SCI (mg/ml) 24—hr urinary excretion of sodium (Na+) was calculated as follows: UNaV (nEq/Z4 h) 2 UNa (nEq/ml) x V (ml/24 h) 24—hr urinary excretion ofNAG and s similarly calculated.

The fractional excretion of Na+ (FEM), or percentage of the filtered sodium that is excreted into the final urine, is a measure of tubular Naheabsorptive function. It was computed as s: FENa (%) =100 x [UNa (HEq/nil) x V (ml/24 h)] / PM (nEq/Inl) X Cc, (ml/24 h) Treatment with anti—Adrenomedullin antibody improved several measures of kidney function on day 7 as compared to vehicle: serum creatinine 101 mg/dL (NT—M) vs 155 mg/dL (vehicle) (Fig. 11), BUN 32.08 mg/dL(NT—M) vs. 52.41 mg/dL (vehicle) (Fig. 12), nous creatinine clearance 934.43 mL/24 h (NT-M) vs. 613.34 mL/24 h (vehicle) (Fig. 13), fractional secretion ofNa+ 0.98 % (NT-M) vs. 1.75 % (vehicle) (Fig. 14).

Exam In the mice CLP model described above, the effect of ent with drenomedullin antibody NT—M on several parameters ofkidney function was igated.

NT—M caused a three- and two—fold higher diuresis and creatinine clearance, respectively, ultimately resulting in lower creatinine, urea, and NGAL blood concentrations at the end of the experiment (see Table 10). Moreover, keratinocyte—derived chemokine (KC) trations in the kidney were significantly lowered by treatment with NTwM (Fig. 15).

Table 10: Parameters of kidney function in the vehicle- (n=ll) and NT—M-treated (n=9) animals. Blood concentrations were measured in samples taken at the end of the experiment.

NGAL = neutrophil gelatinase—associated Iipocalin. All data are median (quartiles).

Vehicle NT-M —Value Urine output [pL'g_1‘h_1] 4.4 (3.5;16.5) 15.2 (13.9;225) 0.033 nine clearance [ills-mind] 197 (110301) 400 (316;509) 0.006 Creatinine[ug-mL']] .l.83(1.52;3.04) l.28(1.20;1.52) 0.010 U‘rea[ug~mL“1] 378 (268513) 175(101;184) NGAL[ug-mL'1] 16 (1520) 11 (10;13) 0.008 The ments were performed as follows: Creatinine, urea, and neutrophil gelatinase-associated lipocalin WGAL) Blood NGAL concentrations were measured using a commercial ELISA (mouse NGAL, RUO 042, BioPorto stics A/S, ka Gentofte). Urea and creatinine trations were measured with a capillary column (Optima—5M8, ey~NageL Diiren, Germany) gas chromatography/mass spectrometry system (Agilent 5890/5970, Boblingen, Germany) using 2H3-creatinine (CDN isotopes, Pointe—Claire, QU, Canada) and methyl-urea (FlukaChemikalic-n, Buchs, Switzerland) as internal rds. After deprcteinization with acetonitrile, centrifugation and evaporation to dryness, the supernatant was reconstituted in formic acid, and extracted over a weak anion exchange column (WCX, Phenomenex, Aschaffenburg, Germany). Acetonitrile plus s(trimcthylsilyl)trifluoroacetamide and N- (tert—butyldimethylsilyl)-N—1nethy1trifluoroacetamide allowed formation of the urea tert-butyl- dimethylsilyl- and the creatininetrimethylsilyl—derivatives, respectively. Ions m/z 231 and 245, and m/z 329 and 332 were monitored for urea and creatinine es and internal standards, respectively. From the urine output and the plasma and urine creatinine concentrations creatinine nce was calculated using the standard formula.

Sample preparation The kidney which was stored at —80°C was ted with a homogenizer in PBS and lysed with a 12-fold concentrated buffer for a whole cell lysate (100 mM Tris pH 7,6; 500 mM NaCl; 6 mM EDTA; 6 mM EGTA; 1 % Triton—X400; 0,5 % NP 40; 10 % Glycerol; Protease- Inhibitors (B—Glycerolphosphate 2 mM; DTT 4 mM; Leupeptine 20 uM; Nattiumorthovanadate 0,2 mM)) and subsequently centrifuged. The whole cell lysate was obtained out of the supernatant; the pellet consisting of cell remnants was discarded. The amount of protein was determined etrically with a cially available protein assay (Bio—Rad, Hercules, CA) and the specimens were adjusted in the way that the final protein concentration was 4 ug/ul. The samples for the Multiplex— and EMSA analysis were diluted 1:1 with EMSA buffer (10 mM Hepes; 50 mM KCl; 10 % Glycerol; 0,1 mM EDTA; 1 mM DTT), the samples for the immuno blots 1:1 with 2—fold Sample Buffer (2 0/0 SDS; 125 mM Tris—HCL (pH 6,8 at 25°C); 10 % Glycerol; 50 mM DTT; 0,01 % Bromophenol blue).

Levels of keratinocyte—derived chemokine (KC) concentrations were determined using a mouse multiplex cytokine kit lex Pro Cytokine Assay, Bio-Rad, Hercules, CA), the assay was performed by using the Bio~plex suspension array system with the manufacturer’s instructions (see also Wagner F, Wagner K, Weber S, Stahl l3, Kndferl MW, Huber—Lang M, Seitz DH, Asfar P, Calzia E, Senftleben U, Gebhard F, Georgieff M, Radermacher P, Hysa V.

Inflammatory effects of hypothermia and inhaled H28 during itated, hyperdynamic murine septic shock. Shock 2011;35:396—402; and Wagner F, Scheuerle A, Weber S, Stahl B, McCook O, Kntiferl MW, Huber~Lang M, Seitz DH, Thomas J, Asfar P, Szabé C, Mdller P, d F, Georgieff M, Calzia E, Radermacher P, Wagner K. Cardiopulmonary, histologic, and inflammatory effects of intravenous NaZS after blunt chest trauma-induced lung ion in mice. J Trauma 2011;71:1659—1667). In brief, the appropriate cytokine standards and samples were added to a filter plate. The samples were incubated with dies chemically attached to fluorescent—labeled micro beads. Thereafter, premixed detection antibodies were added to each well, and subsequently, streptavidin-phycoerythrin was added.

Beads were then pended, and the cytokines on e was quantified using the Bio—Plex protein array reader. Data were automatically processed and ed by Bio—Plex Manager Software 4.1 using the standard curve produced from recombinant cytokine standards. Levels below the detection limit of the assays were set to zero for statistical purposes.

Example 9 In the mice CLP model bed above, the effect of treatment with anti—adrenomedullin antibody NT—M on the liver was investigated.

NT—M caused a significant lowering of keratinocyte—derived chemokine (KC) concentrations in the liver (Fig. 16).

Measurement of keratinocyte—derived ine (KC) was done analogous to e 8 (kidney).

Exam le 10 In the mice CLP model described above, the effect of treatment with anti-adrenomedullin antibody NT—M on several cytokines and chemokinesin the blood circulation a) was investigated.

Cyrokine and chemokine trations Plasma levels of tumor necrosis factor (TNF)—u, interleukin (lL)—6, monocyte chemoattractant protein (MCP)~1, and keratinocyte-derived chemokine (KC) concentrations were determined using a mouse multiplex cytokine kit (Bio-Plex Pro Cytokine Assay, Bio—Rad, Hercules, CA), the assay was performed by using the Bio~plex suspension array system with the manufacturer’s ctions (see also Wagner F, Wagner K, Weber S, Stahl B, Knoferl MW, Huber—Lang M, Seitz DH, Asfar P, Calzia E, Senfileben U, Gebhard F, Georgieff M, acher P, Hysa V. Inflammatory effects of hypothermia and inhaled H28 during resuscitated, hyperdynamic murine septic shock. Shock 2011;35:396—409.; and Wagner F, Scheuerle A, Weber S, Stahl B, McCcok O, Kndferl MW, Huber—Lang M, Seitz DH, Thomas J, Asfar P, Szabo C, Moller P, Gebhard F, Georgieff M, Calzia E, Radermacher P, Wagner K.

Cardiopulmonary, histologic, and inflammatory effects of intravenous Na28 after blunt chest trauma—induced lung contusion in mice. J Trauma 2011;71:1659-1667). In brief, the appropriate cytokine standards and s were added to a filter plate. The samples were incubated with antibodies chemically attached to cent—labeled micro beads. fter, premixed detection antibodies were added to each well, and subsequently, streptavidinm phycoerythrin was added. Beads were then re-suspended, and the cytokines reaction mixture was fied using the Bio—Piex protein array reader. Data were automatically processed and analyzed by Bio—Flex Manager Software 4.1 using the rd curve produced from recombinant cytokine standards. Levels below the detection limit of the assays were set to zero for statistical purposes.

Plasma levels and kidney tissue concentrations of tumor necrosis factor (TNF)-or, interleukin (IL)—6 and IL-10, te chemoattractant protein (MCP)-l, and keratinocyte—dervived chemokine (KC) were determined using a cially available “Multiplex Cytokine Kit” ('Bio-Plex Pro Precision Pro Cytokine Assay, Bio—Rad, Hercules, CA), which allows to collect several parameters out of one single sample. The individual work steps of the assay were performed according to the manufacturer‘s instructions (see also Wagner F, Wagner K, Weber S, Stahl B, Kndferl MW, Huber—Lang M, Seitz DH, Asfar P, Calzia E, Senftleben U, Gebhard F, Georgieff M, Radermacher P, Hysa V. Inflammatory effects of hypothermia and inhaled H23 during resuscitated, hyperdynamic murine septic shock. Shock 2011;35:396—402; and Wagner F, Scheuerle A, Weber S, Stahl B, McCook O, rl MW, Huber—Lang M, Seitz DH, Thomas J, Asfar P, Szabé C, Mdller P, Gebhard F, Georgieff M, Caizia E, Radermacher P, Wagner K. Cardiopulmonary, histologic, and inflammatory effects of intravenous NaZS after blunt chest trauma-induced lung contusion in mice. J Trauma 1:1659—1667).

In brief, the fluorescence—labed pheres s”) were added to a 96—well plate, followed by two washing steps, the addition of al standards and the addition of plasma— and kidney honiogenate samples. During the subsequent incubation the single cytokines bind to the antibodies attached to polystyrene-beads. After the addition of the cytokine—specific —labeled antibodies, which are for the detection of the single cytokines, and an additional incubation time, subsequently phycoerythiin—labeled streptavidine was added. After an additional tion time, beads were then resuspended, and the plates could be measured with a specific flow cytometer (Bio—Flex suspension array system, Bio—Rad, es, CA).

Data were automatically processed and analyzed by Bio—Plex Manager Software 4.1 using the standard curve produced from recombinant cytokine standards. For the plasma levels the concentration was provided in pg * mL‘l, the concentration of the kidney homogenates were ted to the appropriate protein concentration and provided in pg * rng'I n.

NT—M caused a significant lowering of plasma concentrations of IL—6 (Fig. 17), IL—lO (Fig. 18), keratinocyte—derived ine (KC) (Fig. 19), monocyte chemoattractant protein~1 (MCP~1) (Fig. 20), TNF—alpha (Fig. 21).

Example 11 Ischemia/Reperfusion-Induced Acute Kidney Injury Another non-septic acute kidney injury model has been established, where acute kidney injury is induced by ischemia/reperfusion (Nakamoto M, o Jl, Shanley PF, Chan L, and Schrier RW. In Vitro and in vivo protective effect of atriopeptin III on ischemic acute renal failure. J ClinInvest 80:698—705, 1987., Chintala MS, Bernardino V, and Chiu P]S. Cyclic GMP but not cyclic AMP prevents renal platelet accumulation ing ischemia— reperfusion in anesthetized rats. J PharmacolExpTher 271:1203—1208, 1994). This model was used to assess Whether treatment with anti-adrenomedullin antibody can improve kidney function.

The ment was performed as follows: Effect of a NTmM on Acute Kidney Injury Induced by Ischemia/Reperfusion in Rats Study Design: Test Conc Dosage Rats Group Article Route mg/ml nil/kg rug/kg (Male) 1 LR + vehiclea IV 5 NA x 3 8 2 l—R + NT—M 1v 5 x 3" 8 a vehicle; injected enously (i.V.) 5 min before reperfusion on day 0, followed by injections on days 1 and 2. hNT—M at 4 mgfkg was injected intravenously (i.v.) 5 min before reperfusion on day 0, followed by 2 mg/kg iv each on days 1 and 2.

CUrine collection on days ~l, 0, 1 and 2, with blood chemistry and urine analysis on days 0, 1, 2 and 3, respectively. Plasma samples were collected in EDTA tubes (Days 0 iate before surgery), 1, 2: 100 pl], before vehicle or TA; Day 3: 120 iii.

Clinical observations: daily before surgery, following surgery and throughout treatment.

Groups of 8 male Sprague—Dawley rats weighing 250 to 280 g were used. The animals were kept on a 12—hr dark cycle and receive a standard diet with distilled water ad libitum.

The animals e fluid supplements (0.9% NaCl and 5% dextrose/ 1:1, 10 nil/kg 13.0.) 30 min prior to surgery (day 0). The rats were anaesthetized with pentobarbital (50 mg/kg, i.p.).

The abdominal cavity was exposed via a midline on, followed by intravenous administration of heparin (100 U/kg, i.v.) and both renal es were occluded for 45 min by using vascular clamps. Immediately after removal of the renal clips, the kidneys were observed for additional 1 min to ensure color change indicating blood reperfusion. The test compound (NT—M) and vehicle (phosphate buffered saline) were ed intravenously 5 min before usion, followed by daily injection on days 1 and 2.

Urine collection. The 24—h urine tion on ice was initiated at 2411 before ischemia/reperfusion on day -1 (-2411 to Oh), and day 0 (0—24h), day 1 (24-4811) and day 2 (48« 72h) after reperfusion, Blood collection: 0.4 ml blood was collected through the tail vein into EDTA tubes at 011 (before 1 RI surgery), 24h (before e or TA), 48h (before vehicle or TA) and 7211 for determination of plasma creatinine/Na+/K+, and BUN; 2 ml blood was collected through venal cava terminally.

The animals were placed in individual cages where urine was collected for 24 h day —1 (—24h- 011), day 0 (0—2411), day 1 (24-48h) and day 2 h) after reperfusion on day 0. Urine volume, urinary Nar, K+, and creatinine were measured.

The creatinine clearance (CCr) was calculated as follows: CCr (ml/24 h) = [UCr (mg/nil) X V (ml/2411)] / PCr l) The 24—hr urinary ion of sodium (Na+) was calculated as follows: UNaV (qu/24 h) : UNa (qu/ml) x V (ml/24 h) The fractional excretion of Na+ (FENa), or percentage of the filtered sodium that is excreted into the final urine, is a measure of tubular Na+ reabsorptive function. It was computed as follows: FENa (%) =100 x [UNa (qu/rnl) K V (ml/24 h)} / PNa (qu/ml) X CCr (ml/24 h) Treatment with anti—Adrenomedullin antibody improved several measures ofkidney function: Blood urea nitrogen (BUN) showed a strong increase in the e group (0 h: 17.49 mg/clL, 24 h: 9885 mg/dL, 48 11: 109.84 mg/dL, 72 11: 91.88 mg/dL), which was less pronounced with NT—M treatment (0 h: 16.33 mg/dL, 24 h: 84.2 mg/dL, 48 h: 82.61 ing/dL, 72 h: 64.54 nig/dL) (Fig. 22).

Serum creatinine developed similafily: Vehicle group (0 h: 0.6l mgldL, 24 h: 3.3 mg/dL, 48 h: 3.16 mg/dL, 72 h: 2.3] mg/dL), NT—M group: (0 h: 0.59 mg/dL, 24 h: 2.96 mg/dL, 48 h: 2.31 nag/(112,72 h: 1.8 mg/dL) (Fig. 23).

The nous creatinine clearance dropped massively on day one and thereafter improved better in the NT-M group than in the vehicle group. Vehicle group: (0 h: 65.17mL/h, 24 h; 3.5mL/h, 48 h: 12.61mL/h, 72 h: L/h), NT—M group:(0 h: 70.11mL/h, 24 h: 5.84mL/h, 48 h: 21.23mL/h, 72 h: 26.6lmL/h) (Fig. 24).

FIGURE DESCRIPTION Fig. in: Illustration of antibody formats — Fv and scFv~Variants Fig 1!): Illustration of antibody formats —~ heterologous fusions and bifiinctional antibodies Fig 1c: Illustration of antibody formats w bivalental antibodies and bispecific antibodies Fig. 2: hADM l—S2 (SEQ ID No. 21) mADM 1—50 (SEQ ID No. 22) aa 121 n ADM (SEQ ID No. 23) aa 1—42 ofhuman ADM (SEQ ID No. 24) aa 43-52 ofhuman ADM (SEQ ID No. 25) aa 1—] 4 ofhuman ADM (SEQ ID NO: 26) aa l—lO n ADM (SEQ ID NO: 27) aa 16 n ADM (SEQ ID NO: 28) aa 1—32 n mature human ADM (SEQ 1D NO: 29) aa 1-40 ofmature murine ADM (SEQ ID NO: 30) aa 1—31 ofmature murine ADM (SEQ ID NO: 31) Fig. 3: a: Dose response curve of human ADM. Maximal CAMP stimulation was adjusted to 100% activatiou b: Dose/ inhibition curve of human ADM 22—52 (ADM-receptor antagonist) in the presence of 5.63nM hADM. 0: Dose/ inhibition curve of CT—H in the presence of 5.63 nM hADM. (1: Dose/ inhibition curve ofMR~H in the presence of 5.63 11M hADM. 6: Dose/ inhibition curve of NT—H in the presence of 563 nM hADM. f: Dose response curve of mouse ADM. Maximal CAMP ation was adjusted to 100% activation g: Dose/ inhibition curve of human ADM 22-52 (ADM—receptor antagonist) in the ce of 0,67 nM 1nADM. h: Dose/ inhibition curve of CT—M in the presence of 0,67 nM mADM. i: Dose/ inhibition curve ofMR-M in the presence of 0,67 11M mADM. j: Dose/ inhibition curve of NT—M in the presence of 0,67 11M mADM. k: shows the inhibition ofADM by F(ab)2 NT—M and by Fab NT—M 1: shows the inhibition ofADM by F(ab)2 NT—M and by Fab NT—M Fig. 4: This figure shows a typical hADM dose}l signal curve. And an hADM dose signal curve in the presence of 100 ug/mL dy NT—H.

Fig. 5: This figure shows the stability of hADM in human plasma (citrate) in absence and in the presence ofNT—H antibody.

Fig. 6: Alignment of the Fab with homologous human fiamework sequences Fig. 7: This figure shows the enalin requirements for early and late treatment with NT-M Fig. 8: This figure shows urine production after early and late treatment with NT—M Fig. 9: This figure shows the fluid balance after early and late treatment with NT—M Fig. 10: Liver tissue activation of nuclear factor kappa—light—chain gene enhancer in B cells (NFmKB) analyzed by electophoretic mobility shift assay (EMSA). # depicts 1 VS. vehicle.

Fig. 11: Development of serum creatinine overtime. Mean 44— SEM are shown.

Fig. 12: Development ofblood urea nitrogen (BUN) over time. Mean +/- SEM are shown.

Fig. 13: Development of endogenous creatinine clearance over time. Mean +/— SBM are shown.

Fig. 14: Development of fractional secretion ofNa+ over time. Mean +/~ SEM are shown.

Fig. 15: Keratinocyte—derived chemokine (KC) levels ined in relation to the total kidney protein extracted. The white box—plot shows results obtained with vehicle, the grey box-plot shows results obtained afier ent with NT-M.

Fig. 16: Keratinocyte—derived chemokine (KC) levels determined in relation to the total liver n extracted. The white box—plot shows results obtained with e, the grey box-plot shows results obtained after treatment with NT—M.

Fig. 17: Plasma lL—6 levels. The white box—plot shows results obtained with vehicle, the grey box-plot shows results obtained after treatment with NT-M.

Fig. 18: Plasma IL—10 levels. The white box—plot shows s obtained with vehicle, the grey box~ plot shows s obtained after treatment with NT—M.

Fig. 19: Plasma keratinocyte—derived chemokine (KC) levels. The white box—plot shows results obtained with vehicle, the grey box-plot shows results obtained after treatment with NT-M.

Fig. 20: Plasma monocyte chemoattractant protein—l (MCPwl) levels. The white box-plot shows s obtained with vehicle, the grey box—plot shows results obtained after treatment with NT-M.

Fig. 21: Plasma TNF—alpha levels. The white box-plot shows results ed with vehicle, the grey box—plot shows results ed after treatment with NT—M.

Fig. 22: Development ofblood urea nitrogen (BUN) over time. Mean +/— SEM are showu.

Fig. 23: Development of serum nine over time. Mean "H— SEM are shown.

Fig. 24: Development of endogenous creatinine clearance over time. Mean +/- SEM are shown.

Claims (34)

1. The use of an anti-adrenomedullin (ADM) antibody or an anti-adrenomedullin antibody fragment or an anti-ADM non Ig scaffold binding to adrenomedullins, in the manufacture of a medicament for therapy or prevention of SIRS, itis, sepsis or shock in a patient for the reduction of the mortality risk for said patient n said antibody fragment or non Ig scaffold binds to the N-terminal part, aa 1-21, of adrenomedullin:

2. Use according to claim 1, wherein said said antibody or antibody fragment or non Ig scaffold is monospecific.

3. Use according to claim 1 or 2, wherein said antibody or fragment or scaffold exhibits a binding affinity to ADM of at least 10-7 M.

4. Use according to any one of claims 1 to 3, wherein said antibody or nt or scaffold is not ADM-binding-Protein-1; complement factor H.

5. Use according to any one of claims 1 to 4, wherein said antibody or fragment or scaffold recognizes and binds to the N-terminal end, aa 1, of adrenomedullin.

6. Use according to any one of claims 1 to 5, characterized in that said antibody, antibody fragment or non-Ig scaffold does not bind to the inal portion of ADM, being aa 43-52 ADM

7. Use according to any one of claims 1 to 6, n said antibody or fragment or scaffold is an ADM stabilizing dy or fragment or scaffold that enhances the half life, tl/2 half retention time, of adrenomedullin in serum, blood, plasma at least 10 %.

8. Use according to claim 7, n said antibody or fragment or scaffold enhances the half life at least, 50 %.

9. Use ing to claim 8, wherein said antibody or fragment or scaffold enhances the half life > 50 %.

10. Use according to claim 9, wherein said antibody or fragment or scaffold enhances the half life > 100 %.

11. Use according to any one of claims 1 to 10, wherein said antibody or fragment or scaffold blocks the bioactivity of ADM not more than 50 %, wherein the ivity of ADM is determined in human recombinant Adrenomedullin receptor cAMP functional assay.

12. Use according to any one of claims 1 to 11, wherein said patient is an ICU t.

13. Use according to any one of claims 1 to 12, n the shock is septic shock.

14. Use according to any one of claims 1 to 13, wherein said antibody or fragment is a human monoclonal antibody or fragment that binds to ADM or an antibody thereof wherein the heavy chain comprises the sequences

15. Use according to claim 14, wherein said antibody or nt comprises a sequence selected the group comprising.

16. Use according to any one of the claims 1 to 15, n the medicament is to be used in combination with vasopressors and/or fluids administered intravenously.

17. Use according to claim 16, wherein the vasopressor is catecholamine.

18. Use according to any one of the claims 1 to 17, wherein the medicament is to be used in combination with ADM binding protein-1, complement factor H, and/or further active ingredients.

19. Use according to any one of the claims 1 to 18, wherein the medicament comprises a pharmaceutical formulation, which itself comprises the dy or fragment or scaffold.

20. Use according to claim 19 n said pharmaceutical formulation is a on.

21. Use according to claim 20, wherein said solution is a ready-to-use solution.

22. Use according to claim 19, wherein said pharmaceutical formulation is in a freeze-dried state.

23. Use ing to any one of claims 1 to 21, wherein the therapy or prevention is by intramuscular administration.

24. Use according to any one of claims 1 to 21, wherein the therapy or prevention is by intravascular administration.

25. Use according to claim 24, wherein the therapy or prevention is by administration via infusion.

26. Use according to any one of the claims 1 to 25, wherein the therapy or prevention is by systemic administration.

27. Use of a vasopressor in the manufacture of a medicament for therapy or prevention of SIRS, meningitis, sepsis or shock in a patient for the reduction of the mortality risk for said patient, wherein said therapy or prevention comprises administration of an antiadrenomedullin (ADM) antibody or an anti-adrenomedullin antibody fragment or an anti-ADM non Ig scaffold binding to adrenomedullins, in combination with said vasopressor, wherein said antibody fragment or non Ig ld binds to the N-terminal part, aa 1-21, of adrenomedullin:

28. Use according to claim 27, wherein the vasopressor is catecholamine.

29. Use of intravenous fluids in the manufacture of a medicament for therapy or prevention of SIRS, itis, sepsis or shock in a t for the ion of the mortality risk for said patient, wherein said y or prevention comprises administration of an antiadrenomedullin (ADM) antibody or an anti-adrenomedullin antibody fragment or an anti-ADM non Ig scaffold binding to adrenomedullins, in combination with said intravenous fluids, wherein said antibody fragment or non Ig scaffold binds to the N- al part, aa 1-21, of adrenomedullin:

30. Use of ADM binding protein-1 in the manufacture of a medicament for y or prevention of SIRS, meningitis, sepsis or shock in a patient for the reduction of the mortality risk for said patient, wherein said therapy or prevention comprises administration of an anti-adrenomedullin (ADM) antibody or an drenomedullin antibody fragment or an DM non Ig scaffold binding to adrenomedullins in combination with said ADM binding protein-1, wherein said antibody fragment or non Ig scaffold binds to the N-terminal part, aa 1-21, of adrenomedullin:

31. A use according to claim 1, substantially as herein described or exemplified.

32. A use according to claim 27, ntially as herein bed or exemplified.

33. A use according to claim 29, substantially as herein described or exemplified.

34. A use according to claim 30, substantially as herein described or exemplified.