Pharmaceutical combination of peptide-gly and peptidylglycine alpha-amidating monooxygenase (PAM)

Abstract

Subject matter of the present invention is a pharmaceutical combination of Peptide-Gly and Peptidylglycine a-amidating monooxygenase (PAM). Subject matter of the present invention is Peptide-Gly for use in treatment and / or prevention of a disease in a subject, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein Peptide-Gly and / or PAM maybe in modified or unmodified form.

Description

[0001] Pharmaceutical combination of Peptide-Gly and Peptidylglycine a-amidating monooxygenase (PAM)

[0002] Subject matter of the present invention is a pharmaceutical combination of Peptide-Gly and Peptidylglycine a-amidating monooxygenase (PAM). Subject matter of the present invention is Peptide- Gly for use in treatment of a disease in a subject, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein Peptide-Gly and / or PAM may be in modified or unmodified form.

[0003] The present invention relates to surprising techniques in order to enrich and achieve a long-lasting elevation of bioactive peptide hormones, e.g. bio-active Adrenomedullin (bio-ADM), in vivo using nonmodified wildtype biosynthetic precursors of these peptides in combination with its activator peptidylglycine alpha-amidating monooxygenase (PAM). The invention further relates to the techniques for use in a method for the treatment and / or prevention of diseases, especially of cardiovascular, neurodegenerative, edematous and / or inflammatory disorders, and to medicaments comprising the combination of compounds for treatment and / or prevention of diseases, especially of cardiovascular, neurodegenerative, edematous and / or inflammatory disorders.

[0004] State of the Art

[0005] All peptide hormones have a short half-life in human circulation in the range of minutes. Further, all peptide hormones share a biosynthetic mechanism based on the same principles. Both, the biosynthetic principles, and short-half-life related challenges with respect to therapeutic utilization of peptide hormones is explained further using the example of Adrenomedullin (ADM), one of the best studied peptide hormones. In short: The gene product of a peptide hormone encoding gene is an amino acid sequence called preprohormone, that encodes for one or more peptide hormones and undergoes several enzymatic cleavage steps mainly by prohormone convertases (PCs) to release said peptide hormones (Fig. 8). In the first step, the n-terminal signal sequence is cleaved from the preprohormone to result in a prohormone. The prohormone further undergoes partial endoproteolytic cleavage by prohormone convertases (i.e. PC-1, PC-2 and PC-3), as well as post-translational modifications, such as formation of disulfide bonds, as well as removal of dibasic residues by Carboxypeptidase activity (CPE). In some cases, the mature peptide hormone(s) are released after this step. For peptides with a c-terminal glycine residue remaining after CPE activity, an additional enzymatic activation step is necessary: The ascorbate dependent enzyme peptidylglycine alpha-amidating monooxygenase (PAM) recognizes the c-terminal Glycine residue in its substrates (e.g. ADM-Gly) and catalyzes a sequential two-step reaction also referred to as amidation or C-terminal amidation. Thereby glyoxylate, originating from the c-terminal glycine, is released and an amidated carboxyl terminal structure is formed in these peptides, which is essential for the peptide’s activity (summarized in Eggelkraut-Gottanka, R. v et al., Curr. Med. Chem. 11, 2651-2665 (2004)). Examples for peptide hormones requiring the amidation reaction as its final activation step are bioactive Adrenomedullin (bio-ADM), Substance P, Vasopressin, Oxytocin, Amylin, PAMP, alpha-MSH, Calcitonin, Kisspeptin, Neuropeptide Y, Vasoactive intestinal peptide (VIP), Thyroliberin and many more.

[0006] The amidated 52 amino acid peptide hormone bio-ADM, belonging to the ADM / calcitonin gene-related peptide (CGRP) superfamily of peptides (Tsuruda, T. et al., Peptides 111, 47 54 (2019)), is known to be produced in various human organs and tissues, including the heart, adrenal endothelial cells, lungs, kidneys, adipose tissue, and vascular endothelium, which contribute to ADM blood levels (Kato, J. et al., Eur. J. Pharmacol. 764, 140- -148 (2015)). The plasma levels of ADM are in the lower picomolar range. Bio-ADM binds to- and activates the heterodimeric G-protein coupled receptor that consists of CRLR and RAMP 2 or 3 (Calcitonin-receptor- like receptor and receptor activity modifying protein 2 or 3). Activation of the ADM receptor leads to intracellular elevation of adenosine 3', 5'-cyclic monophosphate (cAMP) in the receptor-bearing cells. ADM receptors are present on different cell types in almost all organs including endothelial cells. ADM is thought to be metabolized by degradative enzymes in the plasma or cell membranes, and metalloproteases, such as matrix metalloproteinase 2, but also thermolysin, neutral endopeptidase, and thrombin (Lewis, L. K. et al., 18, 733-739 (1997), Martinez, A. et al., Biochem. J. 383, 413-418 (2004). Further, ADM is predominantly cleared in the lung where ADM-receptors are highly expressed (Gibbons, C, et al., Mol. Endocrinol. 21, 783-796 (2007)).

[0007] Bio-ADM and other amidated peptide-hormones as a target and drug-candidate in therapy were investigated in several preclinical and clinical studies.

[0008] In heart-failure, administration of bio-ADM reduced the size of the necrosis zone in myocardial infarction, apoptosis of cardiac myocytes, pronounced remodeling of the left ventricle (in animals), and aldosterone levels (in animals and humans); improvement in hemodynamic parameters (in both humans and animals) and survival (in animals) (see e.g. Niu, P. et al., Hypertens. Res. 26, 731-736 (2003), and Bdlint, L, et al., Circulation Research, 132(9), 1185-1202, 2023.) Continuous administration of bio- ADM in 7 patients with acute heart failure led to a decrease in mean arterial pressure (MAP), pulmonary artery pressure, systemic and pulmonary vascular resistance, and also led to an increase in cardiac output. In patients with congestive heart failure (HF), infused bio-ADM lowered mean arterial pressure and increased heart rate, but to a lower extent when compared to healthy individuals. Bio-ADM infusion lowered the pulmonary arterial pressure in HF patients, but not in healthy patients. Further, increase in cardiac index was observed, an increase in urine volume and sodium excretion and a decrease in aldosterone levels, both in healthy in patients with HF (reviewed in: Bdlint, Let al., Circulation Research, 132(9), 1185-1202, 2023). A 3-month continuous infusion of GLP-1 in heart-failure prone rats resulted in increased survival and a decreased myocyte apoptosis (Poornima et al., Circulation: Heart Failure. 2008;1:153-160). Further, bio-ADM has several physiological effects, such as vasodilation, angiogenesis, cardioprotection, nephroprotection, anti-oxidation, anti-apoptosis and tissue repair and regeneration. Bio-ADM is involved in blood pressure regulation, bronchodilatation, renal function, hormone secretion, cell growth, differentiation, neurotransmission, and modulation of the immune response. Moreover, ADM plays a crucial role as autocrine factor during proliferation and regeneration of endothelial cells. Additionally, bio-ADM promotes angiogenesis, arteriogenesis, prevents cognitive decline after chronic cerebral hypoperfusion and is therefore considered as therapeutic agent in vascular dementia (reviewed in Garcia, M. A. et al., Expert Opin. Ther. Targets 10, 303- -317 (2006) and Bdlint, L et al., 132(9), 1185-1202, 2023)).

[0009] In infectious diseases, such as sepsis, severe pneumonia or septic shock, bio-ADM concentrations in the blood-stream increase markedly (summarized in Kila. T. et al., Hypertens. Res. 45, 389- -400 (2022)). Several reports in the literature show that ADM is indispensable for an integral endothelial barrier function (Van Tier et al., J. Intern. Med. 289, 792-806 (2021)). Administration of ADM to supra- physiological levels exerts strong anti-edematous and anti-inflammatory functions in a variety of inflammatory conditions in animal experiments including sepsis, acute lung injury and inflammation of the intestine (reviewed in Temmesfeld-Wollbruck, Bet al., 944-951 (2007)). Further, elevated circulating levels of bio-ADM were observed in- and associated with hypertension, renal failure, inflammatory bowel disease (IBD) and pancreatitis (Ashizuka, S et al., Biomedicines 9, (2021)). Despite the elevation of bio-ADM in clinical manifestations of diseases, such as IBD, therapeutic administration of bio-ADM showed positive effects on disease outcome (Eto, T. et al., Nephron 89, 121-134 (2001), Ueda, S. et al. Am. J. Respir. Crit. Care Med. 160, 132-136 (1999)). Additionally, high levels of pro-Adrenomedullin or a fragment thereof (which is not mature ADM-NH2) in the circulation seem to indicate the need of the body to repair the function of the vascular endothelium and the need to support vascular integrity. However, low levels of mature ADM (mature ADM-NH2) indicate, that despite of high levels of pro- ADM the conversion from ADM-Gly to mature ADM (mature ADM-NH2) seems to be disturbed. The terms mature ADM, mature ADM-NH2, ADM-NH2, bio-ADM and amidated ADM are used interchangeably throughout the specification.

[0010] In W02019 / 021600A1 it was disclosed that reduced circulatory levels of bio-ADM were present in- and being predictive for dementia, especially Alzheimer’s Disease. A model of subcortical vascular dementia was reproduced in mice by placing micro coils bilaterally on the common carotid arteries. Using mice overexpressing circulating ADM, the effect of ADM was assessed on cerebral perfusion, cerebral angioarchitecture, oxidative stress, white matter change, cognitive function, and brain levels of cAMP, vascular endothelial growth factor, and basic fibroblast growth factor. This data indicate that ADM promotes arteriogenesis and angiogenesis, inhibits oxidative stress, preserves white matter integrity, and prevents cognitive decline after chronic cerebral hypoperfusion. Thus, ADM may serve as a strategy to tackle subcortical vascular dementia Maki, Tet al., 1122-1128 (2011)). The blood-brain barrier (BBB) is a complex dynamic interface that transduces biomechanical and biochemical signals from the vascular system and the brain and is responsible for maintaining homeostasis of the brain by regulating exchange of water, ions, nutrients, metabolites, neurotransmitters, and other cells (e.g., leukocytes), while limiting entry of potentially toxic xenobiotics in the blood. The BBB is formed, in part, by highly specialized endothelial cells that line brain capillaries. The tight junctions formed by brain microvascular endothelial cells (BMECs) regulate paracellular transport, whereas transcellular transport is regulated by specialized transporters, pumps, and receptors. This barrier regulates transport by transducing signals from the vascular system and the central nervous system. Such as the vascular endothelium in general, BBB in particular is disrupted in several pathological conditions, such as, but not limited to, Stroke, multiple sclerosis, Epilepsy, dementias such as Alzheimer’s disease, amyotrophic lateral sclerosis (ALS), Parkinson’s Disease, Sepsis and severe Sepsis, experimental Colitis, Kidney disorders and others (Demeule, M. et al., Vascul. Pharmacol. 38, 339- -348 (2002), Abbott, etal., Neurobiol. Dis. 37, 13 25 (2010). Daneman, R. Ann. Neurol. 72, 648 672 (2012) and Hernandez, L. et al., Sci. Rep. 12, 4414 (2022).

[0011] Such as in vascular endothelium in general, amidated peptide hormones are regulators of the BBB in particular. Evidence is present showing that amidated hormones, such as but not limited to, bio-ADM, Oxytocin, Vasoactive intestinal Peptide (VIP), mediate the BBB function and regulation and / or have beneficial effects on BBB regeneration during and / or after conditions involving BBB disruption (Momenabadi, S. et al., Neuromolecular Med. 22, 557 571 (2020) and Yang, J et al., J. Stroke Cerebrovasc. Dis. 31, 106160 (2022)) . Additionally, administration of PACAP slowed down AD-like pathology in mice and administration of VIP decreased amyloid-B plaques formation and preserved against brain atrophy in 5xFAD transgenic mice (Korkmaz, O. T. et al., J. Mol. Neurosci. 68, 389-396 (2019) and Rat, D. et al., FASEB J. 25, 3208 (2011)). Therefore, it is desired to increase circulating bio- ADM levels, but also levels of other amidated peptides, such as, but not limited to, VIP and PACAP, in clinical conditions as described above, to facilitate among others the regeneration of the endothelial barrier and / or prevent damage of endothelial barrier.

[0012] In humans, the PAM gene is located at chromosome 5q21.1 having a length of 160 kb containing 25 known exons (Gaier et al., 2014, BMC Endocrine Disorders 14). At least 6 isoforms are known to be generated by alternative splicing (SEQ ID Nos. 41 - 46; SEQ ID Nos. 1 - 6 show the respective prepro- forms).

[0013] The PAM enzyme was found to be expressed at different levels in almost all mammalian cell types, with significant expression in airway epithelium, endothelial cells, ependymal cells in the brain, adult atrium, brain, kidney, pituitary, gastrointestinal tract and reproductive tissues (Chen et al., 2018. Diabetes Obes Metab 20 Suppl 2:64-76; Oldham et al., 1992. Biochem Biophys Res Commun 184(1): 323-29; Schafer et al., 1992. J Neurosci 12(1): 222-34). The precursor protein (1-973 amino acids) of the largest known PAM Isoform 1 (SEQ ID No. 1) encoded by the PAM cDNA is depicted in Figure 9. The N-terminal signal sequence or pre-peptide (amino acids 1-20) assures direction of the nascent PAM polypeptide into the secretory lumen of endoplasmic reticulum and is subsequently cleaved co-translationally. Afterwards the PAM-pro-peptide is processed by the same machinery used for the biosynthesis of integral membrane proteins and secreted proteins including cleavage of the pro-region or pro-peptide (amino acids 21-30), assuring proper folding, disulfide bond formation, phosphorylation and glycosylation (Bousquet-Moore et al. 2010. J Neurosci Res 88(12):2535-45}. The sequences of PAM isoforms 1 to 6 after cleavage of the N-terminal signal sequence or pre-peptide (amino acid 1-20) as well as the pro-region or pro-peptide (amino acids 21 to 30) are given as SEQ ID No. 41 to 46.

[0014] As depicted in Figure 9, the PAM cDNA further encodes two distinct enzymatic activities. The first enzymatic activity is named peptidyl-glycine alpha-hydroxylating monooxygenase (PHM; EC 1.14.17.3), is an enzyme capable of catalyzing the conversion of a C-terminal glycine residue to an alpha hydroxy-glycine. The second activity is named peptidyl-a-hydroxy-glycine alpha-amidating lyase (PAL; EC 4.3.2.5) is an enzyme capable of catalyzing the conversion of an alpha hydroxy-glycine to an alpha-amide with subsequent glyoxylate release. The sequential action of these separate enzymatic activities results in the overall peptidyl-glycine alpha amidating activity. The first enzymatic activity (PHM) is located directly upstream of the pro-region (within amino acids 31-494 of isoform 1 (SEQ ID No. 7 and No. 10)). The second catalytic activity (PAL) is located after exon 16 in isoform 1 within amino acids 495-817 (SEQ ID No. 8).

[0015] As depicted in Figure 9, both activities may be encoded together within one polypeptide as a membranebound protein (isoforms 1, 2, 5, 6; corresponding to SEQ ID No. 41, 42, 45 and 46) as well within one polypeptide as a soluble protein lacking the transmembrane domain (TMD) (isoforms 3 and 4; corresponding to SEQ ID No. 43 and 44). While isoforms 1, 2, 5 and 6 remain in the outer plasma membrane after fusion of secretory vesicles with the plasma membrane with subsequent endocytosis and recycling or degradation, soluble PAM isoforms lacking the TMD (isoforms 3 and 4) (amino acids 864-887) are co-secreted with the peptide-hormones (Wand et al., 1985 Metabolism 34(11): 1044-52}.

[0016] Little is known about the role or influence of PAM in clinical conditions. The presence of alpha- amidating activity in human circulation was initially proved by Wand et al. (Wand, G. S et al., Neuroendocrinology 41, 482-489 (1985)}. They reported no sex differences but some variations of PAM activity in certain disease states: Plasma PAM activities were increased in hypothyroid adults as well as in patients with medullary thyroid carcinoma. The activity of PAM in tissues of medullary thyroid carcinoma, pheochromocytoma and pancreatic islet tumors were shown to be elevated suggesting increased formation of amidated peptides in endocrine tumor tissues (Gether, U et al., Mol. Cell. Endocrinol. 79, 53-63 (1991) and Wand, G. Set al., Neuroendocronology 41, 482-489 (1985)}. Patients suffering from multiple endocrine neoplasia type 1 (MEN-1) and pernicious anemia showed a decreased plasma PAM activity in comparison to healthy control subjects. The presence of amidating activity in human cerebrospinal fluid (CSF) was shown by Wand and colleagues. In patients suffering from Alzheimer’s disease (AD) plasma PAM activities were shown to be unaltered when compared to healthy controls, while CSF PAM activities were significantly decreased in comparison to activities from normal specimen (Wand, G. S. et al., Neurology 37, 1057-1061 (1987)). In addition, in WO2015 / 103594 it was proposed that the presence of PAM-Protein in CSF detected by mass spectrometry of AD-patients was reduced compared to healthy controls. Moreover, bio-ADM, one of the hormones amidated by PAM, was shown to be reduced in patients with prevalent and incident Alzheimer’s disease (W 02019 / 154900). Direct associations of circulating PAM activities were reported to be associated with prediction, diagnosis or progression of AD in W02021170816A1 and WO2021170752A1. PAM activities of patients suffering from multiple sclerosis (MS) were shown to be increased in CSF, with a significant decrease in serum (Tsukamoto, T. et al., Intern. Med. 34, 229-232 (1995) and W02010 / 005387). An association between plasma activity of PAM and type-2 -diabetes was described in (WO2014 / 118634). Further, WO2021170816A1 and WO2021170752A1 show an increase of PAM activity in circulation due to acute conditions such as sepsis and shock. So far, no clinical testing of PAM as a drug candidate has been reported.

[0017] In terms of therapeutic use, the direct application of peptide hormones, especially amidated peptide hormones in their wild-type state turned out to be impracticable due to the short in vivo half-life of the peptide hormones. Clinical utilization of GLP-1 showed to be beneficial in type 2 diabetic patients, resulting in clinically relevant positive outcome. Due to the short half-life of GLP-1, it’s administration is dependent on a continuous infusion, which on the other hand depends on supraphysiological doses and was linked to adverse events (Larsen et al., Diabetes Care 1 August 2001; 24 (8): 1416-1421.)

[0018] Clinical testing of bio-ADM was conducted in cardiovascular indications with a measurable hemodynamic end point such as pulmonary hypertension, hypertension, heart failure and acute myocardial infarction (MCI), IBD and refractory Crohn’s disease (see e.g. Ashizuka, S et al., Dig. Dis. Sci. 61, 872-880 (2016) and Kataoka, Y. et al., J. Cardiovasc. Pharmacol. 56, 413-419 (2010)). Bio- ADM administration showed improvement of clinical symptoms in several studies in patients suffering from the aforementioned conditions. Pharmacodynamic effects comprised among others lowering of systemic and pulmonary arterial blood pressure and increase of cardiac output, as well as reduction of serum inflammatory cytokines and improved mucosal healing in IBD (Troughton, R. W. et al., 588-593 (2000) and Nagaya, N. et al., Peptides 25, 2013-2018 (2004)).

[0019] Thus, the above data indicate that ADM induces favorable hemodynamic, hormonal, and myocardial changes both in animal experimental models and in patients. These effects are likely mediated by the vasodilatory properties of intravascular ADM, although they may be mediated by other mechanisms as well. The limitation of bio-ADM administration is the short-lasting nature of achieved effects with an immediate ceasing after the end of administration. These findings correlated well with the known pharmacokinetic profile of bio-ADM having a relatively short in vivo half-life of approximately 22 minutes (Nagata, Set al., Peptides 121, 170133 (2019)). The nature of bio-ADM restricts the application of wild-type (wt) bio-ADM, especially in CVDs, to continuous intravenous injection or infusion, mostly accounted to the strong vasoactive properties of bio-ADM. Single-injections of higher doses of bio- ADM lead to a significant drop of mean arterial blood pressure (W02013064508A1), but also an infusion of bio-ADM at a rate of 100 ng / kg / min in an LPS induced endotoxemia model led to a decrease of MABP to 64 mmHg bearing the risk of inducing a circulatory shock (Ertmer, C. et al., Crit. Care 11, 1 (2007)). In summary, based on evidence from a wealth of experimental data in animals and clinical trials in man, elevation of bio-ADM to supraphysiological levels might be considered as a target mechanism for the treatment of a variety of disease conditions in man and animals. However, the major limitations of the use of ADM as therapeutic agent are the inconvenient applicability of continuous infusion therapy which precludes its use for most of the potential indications and the potentially limited safety margins with respect to hypotension which may result from bolus administrations of ADM.

[0020] To overcome the aforementioned limitations, several strategies were developed with respect to sustainable elevation of specific peptide hormones in the bloodstream. For GLP-1, several agonist drugs, such as Liraglutide, were developed (Hutchinson et al., J Pept Sci. 2017 Feb;23(2):82-94). Sustainable elevation of bio-ADM can be achieved by its modification, thereby targeting the N-terminus of bio- ADM, since it is unrelated to its biological activity. Administration of Adrecizumab, an N-terminal, non-neutralizing anti -ADM antibody, leads to a sustainable increase of bio-ADM in circulation for up to 15 days, leading to improvement of endothelial function and increasing the chance of a positive outcome in severe sepsis or shock (Deniau, B et al., Expert Opin. Investig. Drugs 30, 95-102 (2021)).

[0021] The application of native wild-type forms of amidated peptide-hormones remains challenging, and therefore, alternative strategies allowing a sustainable utilization of amidated peptide-hormones in their native state are of high unmet clinical need.

[0022] Subject Matter of the Invention

[0023] Subject matter if the present invention is Peptide-Gly for use in treatment of a disease in a subject, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein PAM and / or Peptide-Gly may be in modified or unmodified form.

[0024] According to the present invention PAM may mean PAM (SEQ ID No.: 1) and / or its fragments and / or isoforms (SEQ ID No.: 2-6, 9), and / or catalytic subunits like PHM (SEQ ID No.: 7, 10) and PAL (SEQ ID No.: 8), and / or fragments thereof in unmodified or modified form.

[0025] It is apparent to the skilled person that SEQ ID No.: 1 and SEQ ID No.: 2-6 as shown in the description of the respective sequences herein below are the prepro forms of PAM (containing the pre- and the propeptide) and SEQ ID No. 9 is a pro-form fragment of PAM. The skilled person is readily aware which parts of the sequence of the prepro or pro forms of PAM pertain to signal- or pro-peptides, and which parts pertain to mature PAM. Notwithstanding, the respective mature forms of PAM, which are preferred forms of PAM under the present invention, are shown for sake of completeness in SEQ ID No.: 41 - 46.

[0026] For example, according to the present invention, PAM may mean PAM (SEQ ID No.: 41) and / or its fragments and / or isoforms (SEQ ID No.: 42 - 46, SEQ ID No.: 48), and / or catalytic subunits like PHM (SEQ ID No.: 7, 10) and PAL (SEQ ID No.: 8), and / or fragments thereof in unmodified or modified form. This applies likewise, mutatis mutandis, to the further embodiments of the invention.

[0027] SEQ ID No. 9 is a wildtype pro-fragment of PAM with a C-terminal amino acid extension of two amino acids G (Glycine) and S (serine).

[0028] In embodiments of the present invention, PAM is a recombinant PAM protein according to SEQ ID No.: 47.

[0029] In certain embodiments, PAM is a protein or recombinant protein having at least 85%, particularly at least 90%, more particularly at least 95%, even more particularly at least 99% sequence homology therewith, alternatively wherein sequence homology therewith means a sequence homology of 85% to 100%, preferably of 90% to 100%, or more preferably of 95 to 100% or most preferably of 99% to 100%.

[0030] Peptide-Gly may be selected from, but is not limited to, wildtype ADM-Gly (1-53 ADM) (SEQ ID No.: 14), and / or N-terminal truncated fragments thereof (SEQ ID No.: 17-21, 27), and / or GLP-l-Gly (SEQ ID No.: 39-40) and / or modified ADM-Gly, i.e. modified N-terminal truncated fragments thereof (SEQ ID No.: 17-21, 27) or modified wildtype ADM-Gly (1-53 ADM) / SEQ ID No.: 14) and / or modified GLP-l-Gly (SEQ ID No.: 39-40).

[0031] In one embodiment Peptide-Gly may be modified with PEG, in particular 1-100 kDa PEG. Peptide-Gly may be modified with albumin, and / or Fc-fragments of IgG’s and / or with XTEN.

[0032] Subject matter of the present invention is Peptide-Gly for use in treatment of a disease in a subject, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein Peptide-Gly is Adrenomedullin (ADM)-Gly and wherein ADM-Gly may be in modified or unmodified form.

[0033] An embodiment of the present invention is Peptide-Gly for use in treatment of a disease in a subject, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein Peptide-Gly is GLP-l-Gly and wherein GLP-l-Gly may be in modified or unmodified form. Modifications of PAM and / or Peptide-Gly may be selected from the group comprising:

[0034] • Amino acid manipulation, also referred to as site directed mutagenesis, including the insertion, deletion or alteration of one or more amino acids within the polypeptide amino acid sequence reducing immunogenicity and proteolytic instability in vivo. Thereby alterations of one or more amino acids may lead to enhanced protease resistance in vivo.

[0035] • Conjugation and / or bioconjugation of polypeptides of interest with serum proteins, such as albumins or immunoglobulins or parts of immunoglobulins to produce fusion proteins:

[0036] • Fusion of polypeptides with Albumins, e.g. serum Albumin or recombinant serum Albumin.

[0037] • Non covalent binding to serum Albumin due to a conjugated fatty acid chain to a polypeptide of interest. The binding to albumin therefore happens in vivo after application of the fatty acid conjugated substance and is mediated by the fatty acid chain.

[0038] • Fusion of polypeptides with IgG Fc regions or Transferrin.

[0039] • Post-translational modifications attaching natural or synthetic polymers to the polypeptide of interest. Example for such polymers, but not limited to, are

[0040] ■ PEG, either single-stranded or branched PEG, having varying molecular weights covalently fused to polypeptides of interest.

[0041] ■ XTEN, an unstructured polypeptide, covalently fused to polypeptides of interest. XTEN is a 864 single amino-acid sequence composed of amino-acids Ala, Glu, Gly, Pro, Ser and Thr in a randomized manner. Half-life of an XTEN-fusion protein may be tailored by shortening of the XTEN sequence.

[0042] ■ PAS, which is a peptide polymer consisting of amino acids proline, alanine and serine wit 100- 200 PAS repeats forming the polymer.

[0043] ■ ELP (Elastin-like polypeptides) consisting of Valin-Prolin-Glycine-x-Glycine repeats, naturally found in elastin, wherein x relates to any amino-acid except for Proline. ELP can be covalently attached to a polypeptide of interest.

[0044] ■ HAP, which is a repeated sequence of glycine rich (Gly4Ser)n polypeptide, wherein n is inbetween of 100-200 and is covalently attached to the protein of interest.

[0045] ■ GLK, which is a gelatin-like fusion protein. Thereby GLK is a (Gly-X-Y)n structure, wherein X and Y are any amino-acids except for Cysteine, with n = 60 to 1500. GLK can be covalently attached to a polypeptide of interest.

[0046] ■ Carbohydrates and polysaccharides. Carbohydrates, either branched or linear can be attached to the polypeptide of interest e.g. through in vivo N-Glycosylation. Conjugation with Dextrans, Hydroxyethyls (HES), Heparosan (HEP), Hyaluronic acid (HA) represents the attachment of polysaccharides.

[0047] ■ PSA (Polysialic acid), wherein PSA polymers are covalently attached to the polypeptide of interest. Modifications of PAM and / or Peptide-Gly may also include modifications, which allow PAM and / or Peptide-Gly to act as a prodrug. The term "prodrug" denotes a form or derivative of a compound which is metabolized in vivo, e.g., by biological fluids or enzymes by a subject after administration, into a pharmacologically active form of the compound in order to produce the desired pharmacological effect.

[0048] Prodrugs can thus be viewed as drugs containing specialized non-toxic protective groups used in a transient manner to alter or to eliminate undesirable properties in the parent molecule. The group of the afore mentioned prodrugs may comprise Carrier-linked prodrugs (Carrier prodrugs), Cascade prodrugs and PEG-based carrier prodrugs. A carrier-linked prodrug may be a prodrug that contains a temporary linkage of a given active substance with a transient carrier group that produces improved physicochemical or pharmacokinetic properties and that can be easily removed in vivo, usually by a hydrolytic cleavage, wherein a cascade prodrug is a prodrug for which the cleavage of the carrier group becomes effective only after unmasking an activating group and wherein several examples of PEG- based carrier prodrugs exist, most of them with the need for enzymatic activation of the linker between the active drug and the carrier, mostly initiated by enzymatic hydrolysis. Since esters are cleaved very readily and unpredictably in vivo, direct ester linkers for carrier pro drug have limitations to their usability (J. Rautio et al., Nature Reviews Drug discovery, 2008, 7 255-270).

[0049] A PEG-based carrier prodrug may refer to a carrier-linked prodrug wherein the carrier group is PEG. In terms of this invention a PEG-based carrier prodrug of ADM-Gly may mean the addition of a linker structure according to formula 1 to the c-terminus of ADM-Gly (SEQ ID No.: 14, 17-21, 27), wherein R1 represents a C-terminal amino acid of ADM-Gly according to SEQ ID No.: 14, 17-21, 27 as indicated in formula 1 and the linker is further conjugated with linear or branched PEG of 1-100 kDa at positions R2 and / or R3 of the linker according to formula 1, and wherein n according to formula 1 may be 1, 2 or 3.

[0050] The skilled person will readily recognize in view of the purpose of the present invention that PEG is reasonably attached to the N-terminal end of ADM-Gly. Therefore, embodiments of the invention relating to a PEG-based carrier prodrug of ADM-Gly may mean the addition of a linker structure according to formula 1 to the N-terminus of ADM-Gly (SEQ ID No.: 14, 17-21, 27), wherein R1 represents a N-terminal amino acid of ADM-Gly according to SEQ ID No.: 14, 17-21, 27 as indicated in formula 1 and the linker is further conjugated with linear or branched PEG of 1-100 kDa at positions R2 and / or R3 of the linker according to formula 1, and wherein n according to formula 1 may be 1, 2 or 3.

[0051]

[0052] Formula 1

[0053] The person skilled in the art is aware that also other forms of prodrugs are known in the art. For examples of such prodrugs see: Design of Prodrugs, edited by H. Bundgaard, (Elsevier, 1985) and Methods in Enzymology, Vol. 42, p. 309-396, edited by K. Widder et al. (Academic Press, 1985); A Textbook of Drug Design and Development, edited by Krogsgaard-Larsen and H. Bundgaard, Chapter 5 “Design and Application of Prodrugs” by H. Bundgaard p. 113-191 (1991); H. Bundgaard, Advanced Drug Delivery Reviews 8, 1-38 (1992); H. Bundgaard, et al., Journal of Pharmaceutical Sciences, 77, 285 (1988); and N. Kakeya, et al., Chem. Pharm. Bull., 32, 692 (1984).

[0054] Modified PAM may refer to PAM (SEQ ID No.: 1-10) modified via amino acid manipulations, preferably via fusion to Albumins, e.g. serum Albumin or recombinant serum Albumin, more preferably via non covalent binding to serum Albumin due to a conjugated fatty acid chain to PAM, more preferably via fusion with IgG Fc regions or Transferrin, most preferably via post-translational modifications attaching natural or synthetic polymers, whereas the natural or synthetic polymer to be used is HAP, preferably ELP, more preferably PAS, more preferably PSA, more preferably GLK, more preferably XTEN and most preferably PEG.

[0055] Modified PAM may refer to recombinant PAM (SEQ ID No.: 47) modified via amino acid manipulations, preferably via fusion to Albumins, e.g. serum Albumin or recombinant serum Albumin, more preferably via non covalent binding to serum Albumin due to a conjugated fatty acid chain to PAM, more preferably via fusion with IgG Fc regions or Transferrin, most preferably via post- translational modifications attaching natural or synthetic polymers, whereas the natural or synthetic polymer to be used is HAP, preferably ELP, more preferably PAS, more preferably PSA, more preferably GLK, more preferably XTEN and most preferably PEG. PEG-PAM may refer to PAM (SEQ ID No.: 1-10) modified with 1+n molecules of polyethylene glycol (PEG), whereas n is an integer in the range of 0 to 100 and one PEG molecule has a molecular weight in the range of 1-100 kDa and is either a linear molecule or a branched molecule with b+1 branches, whereas b is an integer in the range of 0 to 20. In specific embodiments PEG-PAM may refer to PAM (SEQ ID No.: 1-10) modified with PEG-5000 (5 kDa PEG), or PEG-10000 (10 kDa PEG).

[0056] In other more preferred embodiments, modification of PAM is modification with PEG (PEGylation), preferably PEG with an average molecular weight of 5-10 kDa (PEG 5000 to PEG 10.000) or, more preferably with an average molecular weight of about 5 kDa (PEG 5000). In further embodiments, modification of PAM with PEG (PEGylation) shall mean the attachment of PEG 5000 or 10000 as detailed herein to serine or lysine side chains of PAM, more preferably the attachment of PEG 10000 as detailed herein to lysine side chains of PAM, or more preferably the attachment of PEG 5000 as detailed herein to lysine or serine, even more preferably to serine side chains of PAM.

[0057] In other more preferred embodiments, modification of PAM is modification with XTEN as defined herein, preferably with XTEN having a sequence of SEQ ID No: 19, more preferably to lysine or serine side chains of PAM, more preferably to lysine side chains of PAM, or alternatively via a Cys amino acid C-terminally added to PAM.

[0058] PAM modified via site directed mutagenesis may refer to PAM (SEQ ID No.: 1-10) with an insertion of n+1 additional amino-acids and / or n+1 deletions of amino-acids and / or n+1 exchanged amino-acids in the given amino-acid sequence (SEQ ID No.: 1-10), wherein n is an integer in the range of 0-100.

[0059] PAM modified via fusion to native serum Albumin or recombinant human serum Albumin may refer to PAM (SEQ ID No.: 1-10) fused to native human serum Albumin or recombinant human serum Albumin at the N-terminus or at the C-terminus of PAM according to (SEQ ID No.: 1-10).

[0060] PAM modified via fusion to IgG Fc regions may refer to PAM (SEQ ID No.: 1-10) fused to human IgGl Fc Region at the N-terminus or at the C-terminus of PAM according to (SEQ ID No.: 1-10).

[0061] PAM modified via XTEN may refer to PAM (SEQ ID No.: 1-10) fused to a XTEN moiety at the N- terminus or at the C-terminus of PAM according to (SEQ ID No.: 1-10) and / or to any surface exposed amino-acid of PAM according to (SEQ ID No.: 1-10).

[0062] PAM modified via XTEN may refer to PAM (SEQ ID No.: 1-10) fused to XTEN moiety (SEQ ID No. 49) at the N-terminus or at the C-terminus of PAM according to (SEQ ID No.: 1-10) and / or to any surface exposed amino-acid of PAM according to (SEQ ID No.: 1-10).

[0063] PEG-PAM may refer to recombinant PAM (SEQ ID No.: 47) modified with 1+n molecules of polyethylene glycol (PEG), whereas n is an integer in the range of 0 to 100 and one PEG molecule has a molecular weight in the range of 1-100 kDa and is either a linear molecule or a branched molecule with b+1 branches, whereas b is an integer in the range of 0 to 20. In a specific embodiment PEG-PAM may refer to recombinant PAM (SEQ ID No.: 47) modified with PEG-5000 (5 kDa PEG), or PEG- 10000 (10 kDa PEG).

[0064] PAM modified via site directed mutagenesis may refer to recombinant PAM (SEQ ID No.: 47) with an insertion of n+1 additional amino-acids and / or n+1 deletions of amino-acids and / or n+1 exchanged amino-acids in the given amino-acid sequence (SEQ ID No.: 47), wherein n is an integer in the range of 0-100.

[0065] PAM modified via fusion to native serum Albumin or recombinant human serum Albumin may refer to recombinant PAM (SEQ ID No.: 47) fused to native human serum Albumin or recombinant human serum Albumin at the N-terminus or at the C-terminus of PAM according to (SEQ ID No.: 47).

[0066] PAM modified via fusion to IgG Fc regions may refer to recombinant PAM (SEQ ID No.: 47) fused to human IgGl Fc Region at the N-terminus or at the C-terminus of PAM according to (SEQ ID No.: 47).

[0067] PAM modified via XTEN may refer to recombinant PAM (SEQ ID No.: 47) fused to a XTEN moiety at the N-terminus or at the C-terminus of PAM according to (SEQ ID No.: 47) and / or to any surface exposed amino-acid of PAM according to (SEQ ID No.: 47).

[0068] PAM modified via XTEN may refer to recombinant PAM (SEQ ID No.: 47) fused to XTEN moiety (SEQ ID No.: 49) at the N-terminus or at the C-terminus of PAM according to (SEQ ID No.: 47) and / or to any surface exposed amino-acid of PAM according to (SEQ ID No.: 47).

[0069] Modified Peptide-Gly may refer to Peptide-Gly modified via amino acid manipulations, preferably via fusion to Albumins, e.g. serum Albumin or recombinant serum Albumin, more preferably via non covalent binding to serum Albumin due to a conjugated fatty acid chain to PAM, mor preferably via fusion with IgG Fc regions or Transferrin, more preferably modified to act as PEG-based carrier prodrug, most preferably via post-translational modifications attaching natural or synthetic polymers, whereas the natural or synthetic polymer to be used is HAP, preferably ELP, more preferably PAS, more preferably PSA, more preferably GLK, more preferably XTEN and most preferably PEG.

[0070] Peptide-Gly modified via PEGylation may refer to Peptide-Gly modified with 1+n molecules of polyethylene glycol (PEG), whereas n is an integer in the range of 0 to 10 and one PEG molecule has a molecular weight in the range of 1-100 kDa and is either a linear molecule or a branched molecule with b+1 branches, whereas b is an integer in the range of 0 to 20.

[0071] Peptide-Gly modified via fusion to native serum Albumin or recombinant human serum Albumin may mean Peptide-Gly fused to native human serum Albumin or recombinant human serum Albumin at the N-terminus of Peptide-Gly.

[0072] Peptide-Gly modified via fusion to IgG Fc regions may refer to Peptide-Gly fused to human IgGl Fc Region at the N-terminus of Peptide-Gly. Peptide-Gly modified via XTEN may refer to Peptide-Gly (SEQ ID No.: 14, 17-21, 27) fused to a XTEN moiety at the N-terminus of Peptide-Gly according to (SEQ ID No.: 14, 17-21, 27).

[0073] Peptide-Gly modified via XTEN preferably relates to Peptide-Gly (SEQ ID No.: 14, 17-21, 27) fused to XTEN moiety (SEQ ID No.: 49) at the N-terminus of Peptide-Gly according to (SEQ ID No.: 14, 17- 21, 27).

[0074] Peptide-Gly modified via XTEN more preferably relates to Peptide-Gly (SEQ ID No.: 14, 17-21, 27, 39, 40) fused to XTEN moiety (SEQ ID No. : 49) at the N-terminus of Peptide-Gly according to (SEQ ID No.: 14, 17-21, 27, 39, 40).

[0075] Fusion may refer to a covalent or non-covalent linkage of two proteins or polypeptides, a protein and a polypeptide, a synthetic polymer and a polypeptide and / or a synthetic polymer and a protein to each other. In the preferred embodiment fusion may refer to covalent linkage.

[0076] A person skilled in the art knows how to create an expression vector for expression of N-terminal or C- terminal fusion proteins. The person skilled in the art knows that fusion of two proteins can be achieved by usage of state-of-the-art techniques to result in a covalent or non-covalent linkage of two proteins, a protein and a polypeptide or a protein and / or a polypeptide and a synthetic polymer. The person skilled in the art knows how to perform site directed mutagenesis.

[0077] In one embodiment Peptide-Gly for use in treatment of a disease in a subject is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) (SEQ ID No.: 1-10) modified via site directed mutagenesis, or PAM (SEQ ID No.: 1-10) modified via fusion to native serum Albumin or recombinant serum Albumin, or PAM (SEQ ID No.: 1-10) modified via fusion to IgG Fc regions, or modified via fusion to XTEN moieties, or PAM (SEQ ID No.: 1-10) modified via PEGylation.

[0078] In a preferred embodiment Peptide-Gly for use in treatment of a disease in a subject is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) (SEQ ID No.: 1-10) modified via PEGylation.

[0079] In another preferred embodiment Peptide-Gly is GLP-l-Gly (SEQ ID No.: 39-40) for use in treatment of a disease in a subject is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) (SEQ ID No.: 1-10) modified via PEGylation.

[0080] In a most preferred embodiment Peptide-Gly is wild-type adrenomedullin-Gly (SEQ ID No.: 14, 17-21, 27) for use in treatment of a disease in a subject is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM) (SEQ ID No.: 1-10) modified via PEGylation.

[0081] In a further embodiment Peptide-Gly modified via fusion to native serum Albumin or recombinant serum Albumin for use in treatment of a disease in a subject is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) (SEQ ID No.: 1-10) modified via site directed mutagenesis, or PAM (SEQ ID No.: 1-10) modified via fusion to native serum Albumin or recombinant serum Albumin, or PAM (SEQ ID No.: 1-10) modified via fusion to IgG Fc regions, or modified via fusion to XTEN moieties, or PAM (SEQ ID No.: 1-10) modified via PEGylation.

[0082] In a preferred embodiment Peptide-Gly modified via fusion to native serum Albumin or recombinant serum Albumin for use in treatment of a disease in a subject is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) (SEQ ID No.: 1-10) modified via PEGylation.

[0083] In another preferred embodiment Peptide-Gly is GLP-l-Gly (SEQ ID No.: 39-40) modified via fusion to native serum Albumin or recombinant serum Albumin for use in treatment of a disease in a subject is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) (SEQ ID No.: 1- 10) modified via PEGylation.

[0084] In the most preferred embodiment Peptide-Gly is adrenomedullin-Gly (SEQ ID No.: 14, 17-21, 27) modified via fusion to native serum Albumin or recombinant serum Albumin for use in treatment of a disease in a subject is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) (SEQ ID No.: 1-10) modified via PEGylation.

[0085] In another embodiment Peptide-Gly modified via fusion to IgG Fc regions for use in treatment of a disease in a subject is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) (SEQ ID No.: 1-10) modified via site directed mutagenesis, or PAM (SEQ ID No.: 1-10) modified via fusion to native serum Albumin or, recombinant serum Albumin, or PAM (SEQ ID No.: 1-10) modified via fusion to IgG Fc regions, or modified via fusion to XTEN moieties, or PAM (SEQ ID No.: 1-10) modified via PEGylation.

[0086] In the preferred embodiment Peptide-Gly modified via fusion to IgG Fc regions for use in treatment of a disease in a subject is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) (SEQ ID No.: 1-10) modified via PEGylation.

[0087] In the more preferred embodiment Peptide-Gly is GLP-l-Gly (SEQ ID No.: 39-40) modified via fusion to IgG Fc regions for use in treatment of a disease in a subject is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) (SEQ ID No.: 1-10) modified via PEGylation.

[0088] In the most preferred embodiment Peptide-Gly is adrenomedullin-Gly (SEQ ID No.: 14, 17-21, 27) modified via fusion to IgG Fc regions for use in treatment of a disease in a subject is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) (SEQ ID No.: 1-10) modified via PEGylation.

[0089] In another embodiment Peptide-Gly modified to act as a PEG-based carrier prodrug for use in treatment of a disease in a subject is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) (SEQ ID No.: 1-10) modified via site directed mutagenesis, or PAM (SEQ ID No.: 1-10) modified via fusion to native serum Albumin or, recombinant serum Albumin, or PAM (SEQ ID No.: 1-10) modified via fusion to IgG Fc regions, or modified via fusion to XTEN moieties, or PAM (SEQ ID No.: 1-10) modified via PEGylation.

[0090] In the preferred embodiment Peptide-Gly modified to act as a PEG-based carrier prodrug for use in treatment of a disease in a subject is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) (SEQ ID No.: 1-10) modified via PEGylation.

[0091] In the more preferred embodiment Peptide-Gly is GLP-l-Gly (SEQ ID No.: 39-40) modified to act as a PEG-based carrier prodrug for use in treatment of a disease in a subject is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) (SEQ ID No.: 1-10) modified via PEGylation.

[0092] In the most preferred embodiment Peptide-Gly is adrenomedullin-Gly (SEQ ID No.: 14, 17-21, 27) modified to act as a PEG-based carrier prodrug for use in treatment of a disease in a subject is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) (SEQ ID No.: 1-10) modified via PEGylation.

[0093] In another embodiment Peptide-Gly modified via fusion to XTEN for use in treatment of a disease in a subject is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) (SEQ ID No.: 1-10) modified via site directed mutagenesis, or PAM (SEQ ID No.: 1-10) modified via fusion to native serum Albumin or recombinant serum Albumin, or PAM (SEQ ID No.: 1-10) modified via fusion to IgG Fc regions, or modified via fusion to XTEN moieties, or PAM (SEQ ID No.: 1-10) modified via PEGylation.

[0094] In the preferred embodiment Peptide-Gly modified via fusion to XTEN for use in treatment of a disease in a subject is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) (SEQ ID No.: 1-10) modified via PEGylation.

[0095] In the more preferred embodiment Peptide-Gly is GLP-l-Gly (SEQ ID No.: 39-40) modified via fusion to XTEN for use in treatment of a disease in a subject is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) (SEQ ID No.: 1-10) modified via PEGylation. In the most preferred embodiment Peptide-Gly is adrenomedullin-Gly (SEQ ID No.: 14, 17-21, 27) modified via fusion to XTEN for use in treatment of a disease in a subject is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) (SEQ ID No.: 1-10) modified viaPEGylation.

[0096] In another embodiment Peptide-Gly modified via PEGylation for use in treatment of a disease in a subject is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) (SEQ ID No.: 1-10) modified via site directed mutagenesis, or PAM (SEQ ID No.: 1-10) modified via fusion to native serum Albumin or recombinant serum Albumin, or PAM (SEQ ID No.: 1-10) modified via fusion to IgG Fc regions, or modified via fusion to XTEN moieties, or PAM (SEQ ID No.: 1-10) modified via PEGylation.

[0097] In the preferred embodiment Peptide-Gly modified via PEGylation for use in treatment of a disease in a subject is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) (SEQ ID No.: 1-10) modified via PEGylation.

[0098] In the more preferred embodiment Peptide-Gly is GLP-l-Gly (SEQ ID No.: 39-40) modified via PEGylation for use in treatment of a disease in a subject is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) (SEQ ID No.: 1-10) modified via PEGylation.

[0099] In the most preferred embodiment Peptide-Gly is adrenomedullin-Gly (SEQ ID No.: 14, 17-21, 27) modified via PEGylation for use in treatment of a disease in a subject is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) (SEQ ID No.: 1-10) modified via PEGylation.

[0100] In another embodiment Peptide-Gly modified to function as PEG-based Prodrug for use in treatment of a disease in a subject is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) (SEQ ID No.: 1-10) modified via site directed mutagenesis, or PAM (SEQ ID No.: 1-10) modified via fusion to native serum Albumin or recombinant serum Albumin, or PAM (SEQ ID No.: 1- 10) modified via fusion to IgG Fc regions, or modified via fusion to XTEN moieties, or PAM (SEQ ID No.: 1-10) modified via PEGylation.

[0101] In the preferred embodiment Peptide-Gly modified to function as PEG-based Prodrug for use in treatment of a disease in a subject is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) (SEQ ID No.: 1-10) modified via PEGylation.

[0102] In the more preferred embodiment Peptide-Gly is GLP-l-Gly (SEQ ID No.: 39-40) modified to function as PEG-based Prodrug for use in treatment of a disease in a subject is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) (SEQ ID No.: 1-10) modified via PEGylation. In the most preferred embodiment Peptide-Gly is adrenomedullin-Gly (SEQ ID No.: 14, 17-21, 27) modified to function as PEG-based Prodrug for use in treatment of a disease in a subject is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) (SEQ ID No.: 1-10) modified via PEGylation.

[0103] In other embodiments Peptide-Gly for use in treatment of a disease in a subject is to be used in combination with recombinant peptidylglycine a-amidating monooxygenase (PAM) (SEQ ID No.: 47) modified via site directed mutagenesis, or recombinant PAM (SEQ ID No.: 47) modified via fusion to native serum Albumin or recombinant serum Albumin, or recombinant PAM (SEQ ID No.: 47) modified via fusion to IgG Fc regions, or modified via fusion to XTEN moieties, or recombinant PAM (SEQ ID No.: 47) modified via PEGylation.

[0104] In preferred embodiments Peptide-Gly for use in treatment of a disease in a subject is to be used in combination with recombinant PAM (SEQ ID No.: 47) modified via PEGylation.

[0105] In other preferred embodiments Peptide-Gly is GLP-l-Gly (SEQ ID No.: 39-40) for use in treatment of a disease in a subject is to be used in combination with recombinant PAM (SEQ ID No.: 47) modified via PEGylation.

[0106] In most preferred embodiments Peptide-Gly is wild-type adrenomedullin-Gly (SEQ ID No.: 14, 17-21, 27) for use in treatment of a disease in a subject is to be used in combination with recombinant PAM (SEQ ID No.: 47) modified via PEGylation.

[0107] In further embodiments Peptide-Gly modified via fusion to native serum Albumin or recombinant serum Albumin for use in treatment of a disease in a subject is to be used in combination with recombinant PAM (SEQ ID No.: 47) modified via site directed mutagenesis, or recombinant PAM (SEQ ID No.: 47) modified via fusion to native serum Albumin or recombinant serum Albumin, or recombinant PAM (SEQ ID No.: 47) modified via fusion to IgG Fc regions, or modified via fusion to XTEN moieties, or recombinant PAM (SEQ ID No.: 47) modified via PEGylation.

[0108] In preferred embodiments Peptide-Gly modified via fusion to native serum Albumin or recombinant serum Albumin for use in treatment of a disease in a subject is to be used in combination with recombinant PAM (SEQ ID No.: 47) modified via PEGylation.

[0109] In other preferred embodiments Peptide-Gly is GLP-l-Gly (SEQ ID No.: 39-40) modified via fusion to native serum Albumin or recombinant serum Albumin for use in treatment of a disease in a subject is to be used in combination with recombinant PAM (SEQ ID No.: 47) modified via PEGylation. In most preferred embodiments Peptide-Gly is adrenomedullin-Gly (SEQ ID No.: 14, 17-21, 27) modified via fusion to native serum Albumin or recombinant serum Albumin for use in treatment of a disease in a subject is to be used in combination with recombinant PAM (SEQ ID No.: 47) modified via PEGylation.

[0110] In other embodiments Peptide-Gly modified via fusion to IgG Fc regions for use in treatment of a disease in a subject is to be used in combination with recombinant PAM (SEQ ID No.: 47) modified via site directed mutagenesis, or recombinant PAM (SEQ ID No.: 47) modified via fusion to native serum Albumin or, recombinant serum Albumin, or recombinant PAM (SEQ ID No.: 47) modified via fusion to IgG Fc regions, or modified via fusion to XTEN moieties, or recombinant PAM (SEQ ID No.: 47) modified via PEGylation.

[0111] In preferred embodiments Peptide-Gly modified via fusion to IgG Fc regions for use in treatment of a disease in a subject is to be used in combination with recombinant PAM (SEQ ID No.: 47) modified via PEGylation.

[0112] In more preferred embodiments Peptide-Gly is GLP-l-Gly (SEQ ID No.: 39-40) modified via fusion to IgG Fc regions for use in treatment of a disease in a subject is to be used in combination with recombinant PAM (SEQ ID No.: 47) modified via PEGylation.

[0113] In most preferred embodiments Peptide-Gly is adrenomedullin-Gly (SEQ ID No.: 14, 17-21, 27) modified via fusion to IgG Fc regions for use in treatment of a disease in a subject is to be used in combination with recombinant PAM (SEQ ID No.: 47) modified via PEGylation.

[0114] In other embodiments Peptide-Gly modified to act as a PEG-based carrier prodrug for use in treatment of a disease in a subject is to be used in combination with recombinant PAM (SEQ ID No.: 47) modified via site directed mutagenesis, or recombinant PAM (SEQ ID No.: 47) modified via fusion to native serum Albumin or, recombinant serum Albumin, or recombinant PAM (SEQ ID No.: 47) modified via fusion to IgG Fc regions, or modified via fusion to XTEN moieties, or recombinant PAM (SEQ ID No.: 47) modified via PEGylation.

[0115] In preferred embodiments Peptide-Gly modified to act as a PEG-based carrier prodrug for use in treatment of a disease in a subject is to be used in combination with recombinant PAM (SEQ ID No.: 47) modified via PEGylation. In more preferred embodiments Peptide-Gly is GLP-l-Gly (SEQ ID No.: 39-40) modified to act as a PEG-based carrier prodrug for use in treatment of a disease in a subject is to be used in combination with recombinant PAM (SEQ ID No.: 47) modified via PEGylation.

[0116] In most preferred embodiments Peptide-Gly is adrenomedullin-Gly (SEQ ID No.: 14, 17-21, 27) modified to act as a PEG-based carrier prodrug for use in treatment of a disease in a subject is to be used in combination with recombinant PAM (SEQ ID No.: 47) modified via PEGylation.

[0117] In other embodiments Peptide-Gly modified via fusion to XTEN for use in treatment of a disease in a subject is to be used in combination with recombinant PAM (SEQ ID No.: 47) modified via site directed mutagenesis, or recombinant PAM (SEQ ID No.: 47) modified via fusion to native serum Albumin or recombinant serum Albumin, or recombinant PAM (SEQ ID No.: 47) modified via fusion to IgG Fc regions, or modified via fusion to XTEN moieties, or recombinant PAM (SEQ ID No.: 47) modified via PEGylation.

[0118] In preferred embodiments Peptide-Gly modified via fusion to XTEN for use in treatment of a disease in a subject is to be used in combination with recombinant PAM (SEQ ID No.: 47) modified via PEGylation.

[0119] In more preferred embodiments Peptide-Gly is GLP-l-Gly (SEQ ID No.: 39-40) modified via fusion to XTEN for use in treatment of a disease in a subject is to be used in combination with recombinant PAM (SEQ ID No.: 47) modified via PEGylation.

[0120] In most preferred embodiments Peptide-Gly is adrenomedullin-Gly (SEQ ID No.: 14, 17-21, 27) modified via fusion to XTEN for use in treatment of a disease in a subject is to be used in combination with recombinant PAM (SEQ ID No.: 47) modified via PEGylation.

[0121] In other embodiments Peptide-Gly modified via PEGylation for use in treatment of a disease in a subject is to be used in combination with recombinant PAM (SEQ ID No.: 47) modified via site directed mutagenesis, or recombinant PAM (SEQ ID No.: 47) modified via fusion to native serum Albumin or recombinant serum Albumin, or recombinant PAM (SEQ ID No.: 47) modified via fusion to IgG Fc regions, or modified via fusion to XTEN moieties, or recombinant PAM (SEQ ID No.: 47) modified via PEGylation.

[0122] In preferred embodiments Peptide-Gly modified via PEGylation for use in treatment of a disease in a subject is to be used in combination with recombinant PAM (SEQ ID No.: 47) modified via PEGylation. In more preferred embodiments Peptide-Gly is GLP-l-Gly (SEQ ID No.: 39-40) modified via PEGylation for use in treatment of a disease in a subject is to be used in combination with recombinant PAM (SEQ ID No.: 47) modified via PEGylation.

[0123] In most preferred embodiments Peptide-Gly is adrenomedullin-Gly (SEQ ID No.: 14, 17-21, 27) modified via PEGylation for use in treatment of a disease in a subject is to be used in combination with recombinant PAM (SEQ ID No.: 47) modified via PEGylation.

[0124] In other embodiments Peptide-Gly modified to function as PEG-based Prodrug for use in treatment of a disease in a subject is to be used in combination with recombinant PAM (SEQ ID No.: 47) modified via site directed mutagenesis, or recombinant PAM (SEQ ID No.: 47) modified via fusion to native serum Albumin or recombinant serum Albumin, or recombinant PAM (SEQ ID No.: 47) modified via fusion to IgG Fc regions, or modified via fusion to XTEN moieties, or recombinant PAM (SEQ ID No.: 47) modified via PEGylation.

[0125] In preferred embodiments Peptide-Gly modified to function as PEG-based Prodrug for use in treatment of a disease in a subject is to be used in combination with recombinant PAM (SEQ ID No.: 47) modified via PEGylation.

[0126] In more preferred embodiments Peptide-Gly is GLP-l-Gly (SEQ ID No.: 39-40) modified to function as PEG-based Prodrug for use in treatment of a disease in a subject is to be used in combination with recombinant PAM (SEQ ID No.: 47) modified via PEGylation.

[0127] In most preferred embodiments Peptide-Gly is adrenomedullin-Gly (SEQ ID No.: 14, 17-21, 27) modified to function as PEG-based Prodrug for use in treatment of a disease in a subject is to be used in combination with recombinant PAM (SEQ ID No.: 47) modified via PEGylation.

[0128] In terms of this invention, the biological activity of the modified compounds shall remain present to an extent of at least 1 % (biological activity 1-300%) when directly compared to the non-modified active moiety, preferably of at least 5% (biological activity 5-300%), more preferably of at least 15% (biological activity 15-300%), more preferably of at least 30% (biological activity 30-300%), more preferably of at least 50% (biological activity 50-300%) more preferably of at least 70% (biological activity 70-300%), more preferably of at least 90% (biological activity 90-300%), more preferably of at least 150% (biological activity 150-300%), more preferably of at least 200% (biological activity 200- 300%) and most preferably of at least 250% (biological activity 250 -300%). In terms of modified PAM, biological activity shall mean the capability of modified PAM to perform c-terminal amidation and may be tested using PAM activity assays with synthetic, labelled tripeptides, such as Tyr-Val-Gly as substrate.

[0129] Preferably the activity is determined as described in Example 2 using wildtype ADM-Gly as substrate. In terms of modified Peptide-Gly, biological activity shall mean the modified peptides capability to stimulate the peptides natural receptor after the modified peptide has been amidated by PAM, which may be tested as described in Example 4 using a distinctive reporter assay.

[0130] Subject matter of the present invention is Peptide-Gly for use in treatment of a disease in a subject, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), wherein said subject is in need of elevated level of corresponding Peptide-NEE in the circulation wherein elevated level means elevation by at least 10 pg / mL, preferably by at least 20 pg / mL, more preferably by at least 30 pg / mL, more preferably by at least 40 pg / mL, most preferably by at least 50 pg / mL.

[0131] An embodiment of the present invention is Peptide-Gly for use in treatment of a disease in a subject, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), wherein said subject is in need of elevated level of corresponding Peptide-NEE in the circulation wherein elevation of the corresponding Pcptidc-N H2may refer to an elevation in the range of 10 - 10000 pg / mL, preferably in the range of 20 - 6000 pg / mL, more preferably in the range of 30- 4000 pg / mL, more preferably in the range of 40-3000 pg / mL, most preferably in the range of 50-1000 pg / mL.

[0132] Another embodiment of the present invention is ADM-Gly for use in treatment of a disease in a subject, wherein ADM-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), wherein said subject is in need of elevated level of Adrenomedullin-NEE (bio-ADM) in the circulation wherein elevated level means elevation by at least 10 pg / mL, preferably by at least 20 pg / mL, more preferably by at least 30 pg / mL, more preferably by at least 40 pg / mL, most preferably by at least 50 pg / mL.

[0133] A further embodiment of the present invention is ADM-Gly for use in treatment of a disease in a subject, wherein ADM-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), wherein said subject is in need of elevated level of Adrenomedullin-NEE (bio-ADM) in the circulation and wherein elevation of bio-ADM may refer to an elevation in the range of 10 - 10000 pg / mL, preferably in the range of 20 - 6000 pg / mL, more preferably in the range of 30- 4000 pg / mL, more preferably in the range of 40-3000 pg / mL, most preferably in the range of 50-1000 pg / mL.

[0134] An embodiment of the present invention is GLP-l-Gly for use in treatment of a disease in a subject, wherein GLP-l-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), wherein said subject is in need of elevated level of amidated GLP-1 (GLP-I-NH2) in the circulation wherein elevated level means elevation by at least 10 pg / mL, preferably by at least 20 pg / mL, more preferably by at least 30 pg / mL, more preferably by at least 40 pg / mL, most preferably by at least 50 pg / mL

[0135] A further embodiment of the present invention is GLP-1 -Gly for use in treatment of a disease in a subject, wherein GLP-1 -Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), wherein said subject is in need of elevated level of GLP-I-NH2 in the circulation and wherein elevation of GLP-1- NH2 may refer to an elevation in the range of 10 - 10000 pg / mL, preferably in the range of 20 - 6000 pg / mL, more preferably in the range of 30 - 4000 pg / mL, more preferably in the range of 40-3000 pg / mL, most preferably in the range of 50-1000 pg / mL.

[0136] In one embodiment Peptide-Gly and Peptide-NLL may be but not limited to, GLP-1 -Gly (SEQ ID No. : 39-40) and GLP-I-NH2, respectively.

[0137] In a preferred embodiment Peptide-Gly and Pcptidc-NIL may be but not limited to, adrenomedullin- Gly (SEQ ID No.: 14, 17-21, 27) and adrenomedullin-NIL (bio-ADM), respectively.

[0138] In one embodiment the elevation of bio-ADM levels may be determined using an immunoassay, that specifically discriminates between amidated and non-amidated ADM, calibrated with wild-type bio- ADM of known concentration and molecular weight. Said assay might be a radioimmunoassay, a fluorescence-based immunoassay or a competitive assay.

[0139] In another preferred embodiment, concentration of bio-ADM may be determined using a luminescencebased sandwich immunoassay as described in Example 1. The molar concentration of the bio-ADM calibrator peptide preparation may be calculated from the mass concentration of the peptide preparation using the molecular weight of bio-ADM, wherein the mass concentration results from dissolving a specific mass of the bio-ADM preparation in a defined volume of a solvent. The molecular weight of the bio-ADM peptide is defined as the sum of weights (in Dalton) of all Atoms in the peptide. Thereby Hydrogen has the weight of 1 Da. The molecular weight of bio-ADM is therefore 6028.9 Dalton.

[0140] Administering the present combination according to the invention leads to a prolonged bioavailability of respective peptide hormone in the combination for at least 30 minutes, preferably for at least 1 hour, more preferably for at least 8 hours, more preferably for at least 12 hours, more preferably for at least 24 hours, more preferably for at least 48 hours, most preferably for at least 72 hours after treatment. Wherein prolonged bioavailability of a peptide hormone may refer to a prolongation in the time-range of 30 minutes-336 hours, preferably in the time-range of 1-288 hours, more preferably in the time-range of 8-240 hours, more preferably in the time-range of 12-216 hours, more preferably in the time-range of 24-192 hours, more preferably in the time-range of 48-180 hours and most preferably in the time-range of 72-168 hours.

[0141] In one embodiment, the bioavailability or prolonged bioavailability of bio-ADM levels may be determined using an immunoassay, that specifically discriminates between amidated and non-amidated ADM, calibrated with wild-type bio-ADM of known concentration and molecular weight, where in bio- ADM levels are determined from blood derived samples drawn at specific time-points after administration of said present combination of the invention. Said assay might be a radioimmunoassay, a fluorescence-based immunoassay or a competitive assay.

[0142] In a preferred embodiment, concentration of bio-ADM may be determined using a luminescence-based sandwich immunoassay as described in Example 1.

[0143] In another embodiment, the bioavailability or prolonged bioavailability of Peptide-Gly may be determined by using an Immunoassay for determination of the total corresponding Peptide concentration irrespective of its amidation status, corresponding to the sum of Peptide-Gly and Pcptidc-NH: concentrations, in the circulation using an antibody combination, that does not react with the c-terminus of corresponding Peptide.

[0144] Afterwards the known concentration of Pcptidc-NFf may be subtracted from the total Peptide concentration to result in the concentration of Peptide-Gly.

[0145] In a preferred embodiment, concentration of Peptide-Gly may be determined using a luminescencebased immunoassay as described in Example 1 for ADM-Gly, wherein the tracer antibody does not cross-react with bio-ADM and specifically reacts with the glycine-extended C-terminus of Peptide-Gly using said preferred assay Peptide-Gly levels may be determined from blood derived samples drawn at specific time-points after administration of said present combination of the invention.

[0146] Specific timepoints may refer to generation of blood-derived samples within of 1 minute-30 minutes, preferably within of 1 minute to 1 hour, more preferably within of 1 minute-8 hours, more preferably within of 1 minute- 12 hours, more preferably within of 1 minute-24 hours, more preferably within of 1 minute-48 hours, more preferably within of 1 minute-72 hours and most preferably within of 1 minute- 336 hours after administration of said present combination of the invention. The assay is calibrated with wild-type ADM-Gly of known concentration and molecular weight. The bioavailability of PAM may be assessed using an activity assay thereby allowing the determination of activity of the enzyme, independent of its molecular weight and therefore independent of any used modification of the enzyme. The activity of an enzyme may be defined in Units or Units / L (Units per Liter of sample), wherein units may mean the amount of product formed by an enzyme in a distinct period of time. In a preferred embodiment the activity is determined from blood derived samples drawn at specific timepoints after administration of said present combination of the invention using an activity assay as described in Example 2.

[0147] Wherein specific timepoints may refer to generation of blood-derived samples within of 1 minute-30 minutes, preferably within of 1 minute to 1 hour, more preferably within of 1 minute-8 hours, more preferably within of 1 minute- 12 hours, more preferably within of 1 minute-24 hours, more preferably within of 1 minute-48 hours, more preferably within of 1 minute-72 hours and most preferably within of 1 minute-336 hours after administration of said present combination of the invention.

[0148] In terms of the present invention, bioavailability may refer to the increase of the concentration or activity of a compound in circulation after administration of said compound or combinations thereof. The term prolonged bioavailability may refer to an increase of the concentration of said compound over a distinct period of time. In terms of the present invention, prolonged bioavailability of Peptide-Gly or Peptide- NH2, may refer to the elevation of concentration of Peptide-NEE or Peptide-Gly in circulation for at least 30 minutes, preferably for at least 1 hour, more preferably for at least 8 hours, more preferably for at least 12 hours, more preferably for at least 24 hours, more preferably for at least 48 hours, most preferably for at least 72 hours after treatment. The term prolonged bioavailability of PAM may refer to the elevation of activity or concentration of PAM in circulation for at least 30 minutes, preferably for at least 1 hour, more preferably for at least 8 hours, more preferably for at least 12 hours, more preferably for at least 24 hours, more preferably for at least 48 hours, most preferably for at least 72 hours after treatment. Wherein prolonged bioavailability of Peptide-NEE and / or Peptide-Gly and / or PAM may refer to a prolongation in the time-range of 30 minutes-336 hours, preferably in the time-range of 1-288 hours, more preferably in the time-range of 8-240 hours, more preferably in the time-range of 12-216 hours, more preferably in the time-range of 24-192 hours, more preferably in the time-range of 48-180 hours and most preferably in the time-range of 72-168 hours.

[0149] Subject matter of the present invention is Peptide-Gly for use in treatment of a disease in a subject, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), wherein said disease is characterized by a level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or a Peptide-Gly / Peptide-amide ratio above a threshold in a sample of bodily fluid of said subject and / or a Peptide-amide below a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or a Peptide-amide below a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof above a threshold and / or a Peptide-amide above a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or a Peptide-amide above a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof above a threshold.

[0150] An embodiment of the present invention is ADM-Gly for use in treatment of a disease in a subject, wherein ADM-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), wherein said disease is characterized by a level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or an ADM-Gly / bio-ADM ratio above a threshold in a sample of bodily fluid of said subject and / or a bio-ADM level below a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or a bio-ADM level below a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof above a threshold and / or a bio-ADM level above a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or a bio-ADM level above a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof above a threshold.

[0151] Another embodiment of the present invention is GLP- 1 -Gly for use in treatment of a disease in a subject, wherein GLP- 1 -Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), wherein said disease is characterized by a level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or a GLP-l-Gly / GLP-I-NH2 ratio above a threshold in a sample of bodily fluid of said subject and / or a GLP-I-NH2 level below a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or a GLP-I-NH2 level below a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof above a threshold and / or a GLP-I-NH2 level above a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or a GLP-I-NH2 level above a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof above a threshold. Peptide-Gly may refer to a c-terminally glycine extended peptide hormone precursor emerging within the biosynthetic pathway of the respective peptide hormone after the exo- and endoproteolytic cleavage of the corresponding prohormone. Peptide-NH2 (equal to Peptide-amide) may refer to an amidated peptide hormone, wherein the c-terminal glycine of its precursor Peptide-Gly is converted by the enzymatic action of the enzyme PAM into an amide moiety thereby producing the active Peptide-NH2 form of the corresponding Peptide-Gly. This means that a given Peptide-Gly has a corresponding Peptide- NH2, wherein the respective amino acid sequence is the same, with the exception of that the C terminal ending has either a NH2 or Gly. Examples include but are not limited to ADM-Gly and GLP-l-Gly, with the corresponding Peptide-amides bio-ADM (ADM- NEE) and GLP-I-NH2, respectively.

[0152] Subject matter of the present invention is Peptide-Gly for use in treatment of a disease in a subject, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein Peptide-Gly is selected from the group comprising from the group of Adrenomedullin family of peptides: Adrenomedullin-Gly, Adrenomedullin- 2-Gly; Intermedin-Short-Gly and Proadrenomedullin N-20 terminal peptide-Gly, from the Bombesin / neurimedin-B / Ranatensin family of peptides: Gastrin-releasing peptide- Gly; Neuromedin-C-Gly; Neuromedin-B-Gly and Nouromedin-B-32-Gly, from the Calcitonin family of peptides: Calcitonin-Gly; Calcitonin gene-related peptide (CGRP) 1-Gly; CGRP 2-Gly and Islet amyloid polypeptide-Gly, from the Chromogranin / Secretogranin family of peptides: Chromogranin-A-Gly; ER-37-Gly; GR-44-Gly and Pancreastatin-Gly, from the FMRF-amide related peptide family of peptides: Neuropeptide AF-Gly; Neuropeptide FF-Gly; Neuropeptide SF-Gly Neuropeptide NPSF-Gly; Neuropeptide RFRP- 1-Gly; Prolactin-releasing peptide PrRP20-Gly and Prolactin-releasing peptide PrRP31-Gly, from the Gastrin / Cholecystokinin family of peptides: Cholecystokinin-Gly; Cholecystokinin- 12-Gly; Cholecystokinin- 18-Gly; Cholecystokinin-25 -Gly; Cholecystokinin-33 -Gly;

[0153] Cholecystokinin-39-Gly; Cholecystokinin-5 -Gly; Cholecystokinin-58-Gly; Cholecystokinin- 8-Gly; Cholecystokinin-7-Gly; Big-gastrin-Gly; Gastrin-Gly; Gastrin- 14-Gly; Gastrin-52- Gly; Gastrin-6-Gly and Gastrin-71 -Gly, from the Glucagon family of peptides: Glucagon-like peptide (GLP) 1-Gly; GLP-l-Gly (7- 37); Pituitary adenylate cyclase-activating polypeptide (PACP) 27-Gly; PACAP 38-Gly; Secretin-Gly; Somatoliberin-Gly; Intestinal Peptide PHM-27-Gly; Intestinal Peptide PHV-42- Gly and Vasoactive intestinal Peptide (VIP)-Gly, from the gonadotropin-releasing hormone family of peptides: Ginadoliberin-l-Gly; Gonadoliberin-2-Gly; Progonadoliberin-l-Gly and Progonadoliberin-2-Gly, from the Kisspeptin family of peptides: Kisspeptin-10-Gly; Kisspeptin-13-Gly; Kisspeptin- 14-Gly; Metastin-Gly and Metastasis-suppressor KISS-l-Gly, from the Melanin-concentrating hormone family of peptides: Neurpeptide-glutamic acid- isoleucine-Gly and Pro-MCH-Gly, from the Neuromedin family of peptides: Neuromedin-S-Gly and Neuromedin-U-25-Gly, from the Neuropeptide-Y (NPY) family of peptides: Neuropeptide Y-Gly; Pancreatic hormone-Gly; Peptide YY-Gly and Peptide YY( 3-36)-Gly, from the opioid family of peptides: Deltrophin I-Gly, from the Orexin family of peptides: Orexin-A-Gly and Orexin-B-Gly, from the Proopiomelanocortin (POMC) family of peptides: Corticotropin-Gly; alpha- Melanotropin-Gly; gamma-Melanotropin-Gly and N-Terminal peptide of proopiomelanocortin-Gly (NPP-Gly), from the RFamide neuropeptide family of peptides: Orexigenic neuropeptide QRF-Gly (QRF- Gly), from the Sauvagine / corticotropin-releasing factor / urotensin I family of peptides: Urocortin- Gly and Urocortin-3-Gly, from the Tachykinin family of peptides: Substance P-Gly (RPKPQQFFGLMG) and Substance P-Gly (RPKPQFFGLMG); Neurokinin A-Gly; Neuropeptide K-Gly and Neurokinin-B-Gly. from the Thyrotropin Releasing Hormone (TRH) family of peptides: pro-TRH-Gly and TRH- Gly, from the Vasopressin / Oxytocin family of peptides: Oxytocin-Gly and Arg-Vasopressin-Gly, from the VGF nerve growth factor inducible protein (VGF) family of peptides: Antimicrobial peptide VGF-Gly (aa554-578) and Neurosecretory protein VGF-Gly. In a preferred embodiment according to the present invention Peptide-Gly are selected from Adrenomedullin-2-Gly (SEQ ID No.: 29), Vasopressin-Gly (SEQ ID No.: 30), Islet amyloid polypeptide-Gly (SEQ ID No.: 31), Substance P-Gly (SEQ ID No.: 32-33),

[0154] Oxytocin-Gly (SEQ ID No.: 34), Vasoactive intestinal peptide-Gly (VIP-Gly) (SEQ ID No.: 35), Neuropeptide Y-Gly (NPY-Gly) (SEQ ID No.: 36), PACAP-Gly (SEQ ID No.: 37-38), GLP-l-Gly (SEQ ID No.: 39-40) and Adrenomedullin-Gly (SEQ ID No.: 14, 17-21, 27).

[0155] In a more preferred embodiment according to the present invention Peptide-Gly are selected from Vasopressin-Gly (SEQ ID No.: 30), Substance P-Gly (SEQ ID No.: 32-33), Oxytocin-Gly (SEQ ID No.: 34), Vasoactive intestinal peptide-Gly (VIP-Gly) (SEQ ID No.: 35), Neuropeptide Y-Gly (NPY-Gly) (SEQ ID No.: 36), PACAP-Gly (SEQ ID No.: 37-38), GLP-l-Gly (SEQ ID No.: 39-40) and Adrenomedullin-Gly (SEQ ID No.: 14, 17-21, 27).

[0156] In the more preferred embodiment according to the present invention Peptide-Gly are selected from Substance P-Gly (SEQ ID No.: 32-33), Oxytocin-Gly (SEQ ID No.: 34), Vasoactive intestinal peptide- Gly (VIP-Gly) (SEQ ID No.: 35), Neuropeptide Y-Gly (NPY-Gly) (SEQ ID No.: 36), PACAP-Gly (SEQ ID No.: 37-38), GLP-l-Gly (SEQ ID No.: 39-40) and Adrenomedullin-Gly (SEQ ID No.: 14, 17- 21, 27).

[0157] In the most preferred embodiment according to the present invention Peptide-Gly are selected from, Vasoactive intestinal peptide-Gly (VIP-Gly) (SEQ ID No.: 35), Neuropeptide Y-Gly (NPY-Gly) (SEQ ID No.: 36), PACAP-Gly (SEQ ID No.: 37-38) and Adrenomedullin-Gly (SEQ ID No.: 14, 17-21, 27).

[0158] In a specific embodiment according to the present invention Peptide-Gly is Adrenomedullin-Gly (SEQ ID No.: 14, 17-21, 27) and / or GLP-l-Gly (SEQ ID No.: 39-40).

[0159] Subject matter according to the invention is Peptide-Gly for use in treatment of a disease in a subject, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM, SEQ ID No.: 1-10), and wherein said disease is selected from the group comprising cardiovascular diseases, kidney / water electrolyte system diseases, brain and neurological diseases, especially dementia, genitourinary disease, gastrointestinal diseases, orthopedic disease, endocrine and metabolic disorders, respiratory disorders, immune disorders and / or inflammatory or infectious diseases.

[0160] Other embodiments of the invention relate to Peptide-Gly for use in treatment of a disease in a subject, wherein Peptide-Gly is to be used in combination with recombinant PAM (SEQ ID 47), and wherein said disease is selected from the group comprising cardiovascular diseases, kidney / water electrolyte system diseases, brain and neurological diseases, especially dementia, genitourinary disease, gastrointestinal diseases, orthopedic disease, endocrine and metabolic disorders, respiratory disorders, immune disorders and / or inflammatory or infectious diseases.

[0161] Subject matter of the present invention is Peptide-Gly for use in treatment of a disease in a subject, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), and wherein said disease is selected from the group comprising pulmonary hypertension and hypertension, type-2 diabetes, inflammatory bowel disease and / or Crohn’s disease, sepsis and / or septic shock, cerebral infarction, mild cognitive impairment (MCI) and Alzheimer’s Disease (AD).

[0162] A preferred embodiment of the present invention is ADM-Gly for use in treatment of a disease in a subject, wherein ADM-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), and wherein said disease is selected from the group comprising pulmonary hypertension and hypertension, type-2 diabetes, inflammatory bowel disease and / or Crohn's disease, sepsis and / or septic shock, cerebral infarction, mild cognitive impairment (MCI) and Alzheimer’s Disease (AD).

[0163] Another preferred embodiment of the present invention is also GLP-l-Gly for use in treatment of a disease in a subject, wherein GLP-l-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), and wherein said disease is selected from the group comprising pulmonary hypertension and hypertension, type-2 diabetes, inflammatory bowel disease and / or Crohn's disease, sepsis and / or septic shock, cerebral infarction, mild cognitive impairment (MCI) and Alzheimer’s Disease (AD).

[0164] Subject matter of the present invention is Peptide-Gly for use in treatment of a disease in a subject, wherein Peptide-Gly is GLP-l-Gly (SEQ ID No.: 39-40), wherein GLP-l-Gly (SEQ ID No.:39-40) is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), and wherein said disease is type-2 diabetes.

[0165] An embodiment of the present invention is ADM-Gly for use in treatment of a disease in a subject, wherein ADM-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), and wherein said disease is type-2 diabetes.

[0166] The blood-brain barrier (BBB) is a continuous endothelial membrane within brain microvessels that has sealed cell-to-cell contacts and is sheathed by mural vascular cells and perivascular astrocyte end-feet. There is an extensive body of evidence from the literature which shows that bio-ADM is indispensable for an intact endothelial and blood-brain barrier function and that administration of bio-ADM and other peptide hormones to supra-physiological levels has protective effects towards endothelial and bloodbrain barrier function (for review Kis B et al.,. Peptides. 2006 Jan;27(l):211-22) The BBB protects neurons from factors present in the systemic circulation, and maintains the highly regulated CNS internal milieu, which is required for proper synaptic and neuronal functioning. BBB disruption allows influx into the brain of neurotoxic blood-derived debris, cells, and microbial pathogens, and is associated with inflammatory and immune responses, which can initiate multiple pathways of neurodegeneration {Sweeney, M. D. et al., Nat. Rev. Neurol. 14, 133-150 (2018)). BBB dysfunction occurs in several diseases, including MS, epilepsy, stroke and dementia, including Alzheimer’s disease (AD). In these conditions, BBB dysfunction plays a central role in the pathology, as BBB disruption leads to ion dysregulation, edema, neuroinflammation, culminating in neuronal dysfunction, increased intracranial pressure and neuronal degeneration (profaci nature).

[0167] The pharmaceutical composition according to the present invention is capable of treating and / or preventing BBB dysfunction and dysregulation as outlined in Example 7 and therefore may be used for treatment and / or prevention of diseases associated with a BBB dysfunction and / or disruption, which specifically include, but are not limited to, brain and neurological diseases such as cerebral infarction, mild cognitive impairment, dementia, cerebrovascular dementia, Alzheimer's disease, and encephalitis.

[0168] The impairment and / or disruption and / or increased permeability of BBB may be quantified in a patient using advanced clinical imaging technologies including, but not limited to, Dynamic Contrast-Enhanced MRI (DCE-MRI), Dynamic Susceptibility Contrast Perfusion Imaging (DSC-MRI), Glucose Chemical Exchange Saturation Transfer Imaging (GlucoCEST), Arterial Spin Labelling (ASL), Intravoxel Incoherent Motion Imaging (IVIM). The advanced clinical imaging techniques may help identify patients with impaired BBB in clinical settings including, but not limited to, acute stroke, Cerebral small vessels disease and dementias, such as mild cognitive impairment (MCI) and Alzheimer’s Disease (AD) (reviewed in: Chassidim, Y. etal., Fluids Barriers CNS 10, 9 (2013) and Elschot EP etal., Invest Radiol. 2021 Jan;56(l)).

[0169] The major component of BBB is the endothelium involved not only in the formation of BBB in particular, but also in the formation of the vascular barrier in general. Blood-brain barrier and vascular barrier dysfunction or endothelial dysfunction is a systemic pathological state of the endothelium and can be broadly defined as an imbalance between vasodilating and vasoconstricting substances produced by and / or acting on the endothelium Deanfield J et al., J Hypertens. 2005 Jan;23(l):7-17.) Normal functions of endothelial cells include mediation of coagulation, platelet adhesion, immune function and control of volume and electrolyte content of the intravascular and extravascular spaces. The endothelium is a cellular monolayer that lines the entire cardiovascular system and regulates many processes including vascular tone, thrombosis, angiogenesis, and inflammation. Endothelial cells have been shown to be phenotypically dynamic and, in response to a variety of local and systemic stimuli, are able to transition between quiescent and activated states {Colombo PC et al., Curr Heart Fail Rep. 2015 Jun;12(3):215-22).). In recent years, emerging research has demonstrated that endothelial dysfunction is a major contributor to cardiovascular disease, including hypertension, atherosclerosis, and congestive heart failure (Gutierrez E, et al., Eur Heart J. 2013 Nov; 34(41): 3175-81)). The endothelium tightly controls the exchange of fluid from the circulation to the surrounding tissues and dysfunction of this barrier leads to uncontrolled fluid extravasation that may result in congestion and / or edema. A common feature of edema (e.g. pulmonary edema) is increased permeability to water low molecular weight solutes (Rocker GM, et al., Thorax. 1987 Aug; 42(8): 620-3).). Endothelial dysfunction can result from and / or contribute to several disease processes, as occurs in hypertension, hypercholesterolaemia, diabetes or septic shock. Endothelial dysfunction is a major pathophysiological mechanism that leads towards coronary artery disease, and other atherosclerotic diseases. Therefore, the pharmaceutical composition of this invention may be used for treatment and / or prevention of diseases associated with an endothelial and / or vascular dysfunction and / or disruption.

[0170] Embodiments of the present invention refer to Peptide-Gly for use in treatment and / or prevention of a disease in a subject, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), and wherein said disease is associated with endothelial barrier dysfunction and / or blood-brain barrier dysfunction.

[0171] The pharmaceutical composition of this invention containing the compound of one aspect of the present invention, a pharmaceutically acceptable salt thereof, or a solvate thereof as an active ingredient has various symptoms prevented or treated by Peptide-NH2, preferably by bio-ADM. Diseases and / or disorders can be prevented or treated as well. The symptoms, diseases and / or disorders include, but are not limited to, the following:

[0172] (1) Brain and neurological diseases: cerebral infarction, mild cognitive impairment, dementia, cerebrovascular dementia, Alzheimer's disease, and encephalitis.

[0173] (2) Infectious diseases caused by infectious organisms such as bacteria, viruses, fungi or parasites, more particularly by infectious bacteria, particularly diseases selected from the group comprising SIRS, sepsis, and septic shock.

[0174] (3) Gastrointestinal diseases: inflammatory diseases (e.g., inflammatory bowel disease or Crohn's disease), ulcerative diseases (e.g., ulcerative colitis), intestinal Behcet's disease, hepatitis, liver fibrosis, cirrhosis, and liver failure. (4) Endocrine and metabolic disorders: diabetes and diabetic organ disorders (e.g., diabetic nephropathy or diabetic retinopathy).

[0175] (5) Cardiovascular disease: heart failure, pulmonary hypertension, arteriosclerosis obliterans, Buerger's disease, myocardial infarction, lymphedema, Kawasaki disease, myocarditis, arrhythmia (for example, arrhythmia after catheter ablation surgery), atrial fibrillation, Aortitis, pulmonary hypertension, hypertension, organ damage due to hypertension, peripheral vascular disease, and arteriosclerosis. (6) Brain / neurological disorders prevented or treated by the pharmaceutical composition according to an embodiment of the present invention is dementia, in particular mild cognitive impairment, cerebral infarction dementia or Alzheimer’s disease.

[0176] Dementia is a clinical syndrome characterized by a cluster of symptoms and signs manifested by difficulties in memory, disturbances in language, psychological and psychiatric changes, and impairments in activities of daily living. The different causes (sometimes referred to as subtyping) of dementia syndrome are: Alzheimer’s disease (about 50% of cases), vascular dementia (about 25%), mixed Alzheimer’s disease and vascular dementia (included in the above, 25%), Lewy body dementia (15%) and others (about 5% combined) including frontotemporal dementia, focal dementias (such as progressive aphasia), subcortical dementias (such as Parkinson’s disease dementia), and secondary causes of dementia syndrome (such as intracranial lesions).

[0177] Alzheimer's disease (AD) is the most prevalent form of dementia. Key molecular mechanisms and histopathological hallmarks in the AD brain comprise a dynamic cascade of biochemical events including the pathological amyloidogenic cleavage of the amyloid precursor protein (APP), the generation of various beta-amyloid species including the amyloid-beta peptide (AP1.42), dimers, trimers, oligomers and subsequent amyloid aggregation and deposition in plaques, abnormal hyperphosphorylation and aggregation of tau protein, progressive intracellular neurofibrillary degeneration, changes within the innate immune system and inflammation and a breakdown of the blood-brain barrier.

[0178] Mild cognitive impairment (MCI) is a heterogeneous clinical condition with several underlying causes. However, the large proportion of MCI represents a transitional state between healthy aging and very mild AD (DeCarli 2003. Lancet Neurol. 2:15-21}. Accordingly, studies suggest that MCI subjects tend to progress to clinically probable AD at a rate of approximately 10%— 15% per year (Markesbery 2010. J Alzheimers Dis. 19:221-228}. A breakdown of the blood-brain barrier in the hippocampus of MCI patients adds evidence to the hypothesis of BBB breakdown to precede neurodegeneration.

[0179] In a preferred embodiment the patient group of Alzheimer's patients may be determined by risks factors such as the occurrence of MCI, the presence of the genetic risk factor ApoE4, as well as age. In a more preferred embodiment the risk factor age of Alzheimer' s patients is defined as an age of at least 60 years.

[0180] Dementia with Lewy bodies (DLB) is a type of dementia that worsens over time. DLB is associated with BBB dysfunction and microvascular lesions (Janelidze S et al., Neurobiol Aging. 2017 Mar;51:104- 112.}. Additional symptoms may include fluctuations in alertness, visual hallucinations, slowness of movement, trouble walking, and rigidity. DLB is the most common cause of dementia after Alzheimer’s disease and vascular dementia. It typically begins after the age of 50. The underlying mechanism involves the formation of Lewy bodies in neurons, consisting of alpha-synuclein protein. A diagnosis may be suspected based on symptoms, with blood tests and medical imaging done to rule out other possible causes. At present no cure for DLB exists. For review see McKeiih et al. 2017. Neurology 89: 88-100.

[0181] Vascular dementia (VaD), also known as multi-infarct dementia (MID) and vascular cognitive impairment (VCI), is dementia caused by problems in the supply of blood to the brain, typically a series of minor strokes, leading to worsening cognitive decline that occurs step by step. The term refers to a syndrome consisting of a complex interaction of cerebrovascular disease and risk factors that lead to changes in the brain structures due to strokes and lesions and resulting in disruption of the blood-brain barrier and changes in cognition. The temporal relationship between a stroke and cognitive deficits is needed to make the diagnosis.

[0182] Frontotemporal dementia (FTD) is the clinical presentation of frontotemporal lobar degeneration, which is characterized by progressive neuronal loss predominantly involving the frontal or temporal lobes, and typical loss of over 70% of spindle neurons, while other neuron types remain intact. FTD is associated with BBB dysfunction and microvascular lesions (Janelidze S, et al., Neurobiol Aging. 2017 Mar;51:104-112).aaA accounts for 20% of young-onset dementia cases. Signs and symptoms typically manifest in late adulthood, more commonly between the ages of 55 and 65, approximately equally affecting men and women. Common signs and symptoms include significant changes in social and personal behavior, apathy, blunting of emotions, and deficits in both expressive and receptive language. Currently, there is no cure for FTD, but there are treatments that help alleviate symptoms. For review see Bott et al. 2014. Neurodegener Dis Manag 4(6): 439-454.

[0183] Differentiating the different dementia syndromes can be challenging, due to the frequently overlapping clinical features and related underlying pathology. In particular, Alzheimer’s dementia often co-occurs with vascular dementia. People with vascular dementia present with progressive cognitive impairment, acutely or sub-acutely as in mild cognitive impairment, frequently stepwise, after multiple cerebrovascular events (strokes). For review see Venkat et al. 2015. Exp Neurol 272: 97-108. Nevertheless, a pathology that is common for all forms of dementia is an impairment of the blood-brain barrier.

[0184] As used herein, “dementia " refers to substantially the clinical syndrome characterized by a cluster of symptoms and signs manifested by difficulties in memory, disturbances in language, psychological and psychiatric changes, and impairments in activities of daily living as outlined above have a common denominator of BBB disruption and / or dysregulation. Therefore “dementia” may refer to Frontotemporal dementia, vascular dementia, Lewy body dementia, Alzheimer’s Dementia and Mild Cognitive Impairment and the pharmaceutical composition according to the present invention may be used for treatment and / or prevention of dementia by protecting the BBB from dysregulation as shown e.g. in Example 7.

[0185] (7) Infectious diseases prevented or treated by the pharmaceutical composition according to an embodiment of the present invention are, in particular, sepsis or septic shock.

[0186] The endothelium is an active contributor to sepsis and as such represents a major target for therapy. During sepsis, endothelial cells amplify the immune response and activate the coagulation system. They are both a target and source of inflammation and serve as a link between local and systemic immune responses. In response to cytokines produced by immune cells, the endothelium expresses adhesion molecules and produces vasoactive compounds, inflammatory cytokines, and chemoattractants (Dolmatova EV et al., Cardiovasc Res. 2021 Jan l;117(l):60-73). Hemodynamic instability plays an important role in the development of sepsis and / or septic shock and arises due to a combination of sepsis- induced vasodilation and vascular leakage, the latter of which is caused by disrupted endothelial integrity. Extensive changes occur in the endothelium as a result of circulating damage-associated molecular patterns and pathogen-associated molecular patterns that activate inflammatory and coagulation pathways during sepsis. In turn, this can result in increased leukocyte adhesion, a procoagulant state, vasodilation and endothelial cell permeability, and ultimately widespread edema, shock and lethal organ dysfunction (Geven C. et al., Shock. 2018 Dec;50(6):648-654.)' . As shown in example 7, the pharmaceutical composition according to the present invention may be used for protecting the endothelial barrier from dysregulation and / or prevent endothelial permeability.

[0187] (8) The gastrointestinal diseases prevented or treated by the pharmaceutical composition according to an embodiment of the present invention are, in particular, inflammatory diseases (e.g., inflammatory bowel disease (IBD) or Crohn's disease), ulcerative diseases (e.g., ulcerative colitis) or intestinal Bechet's disease.

[0188] Inflammatory bowel diseases (IBD) include Crohn’s disease (CD), ulcerative colitis (UC) (and indeterminate colitis), which share several inflammatory characteristics with other chronic immune disturbances including immune activation, leukocyte infdtration into tissues and increased vascular density. The maintenance of normal vascular barrier supports nutrient and 02 exchange, osmotic balance and leukocyte abundance in the extracellular compartment. In IBD, increased vascular permeability leads to tissue edema and damage in both human IBD and animal models of IBD.

[0189] This alteration in solute permeability of the vasculature is not restricted to the gut microcirculation but is widespread affecting the vasculature of other organs including the brain. (Cromer WE et al., World J Gastroenterol. 2011 Feb 7;17(5):578-93). The pharmaceutical composition according to the present invention may be used for protecting the vascular barrier from dysregulation and / or prevent vascular permeability as shown in example 7.

[0190] (9) The endocrine and metabolic disorders prevented or treated by the pharmaceuticals of this embodiment are, in particular, diabetes and diabetic organ disorders (e.g., diabetic nephropathy or diabetic retinopathy).

[0191] Glucagon-like peptide- 1 (GLP-l)-based therapy of type 2 diabetes is executed either by GLP-1 receptor agonists, such as Liraglutide, Albiglutide or Taspoglutide, which stimulate the GLP-1 receptors, or by dipeptidyl peptidase-4 (DPP -4) inhibitors, which prevent the inactivation of endogenous GLP-1 thereby increasing the concentration of endogenous active GLP-1. GLP-1 and its analogues activate pancreatic receptors resulting in improved glycemia through glucose-dependent stimulation of insulin secretion and inhibition of glucagon secretion. There is also a potential beta cell preservation effect, as judged from rodent studies. GLP-1 receptors are additionally expressed in extrapancreatic tissue, having potential for the treatment to reduce body weight and to potentially have beneficial cardio- and endothelioprotective effects. Clinical trials in subjects with type 2 diabetes have shown that in periods of 12 weeks or more, GLP-1 analogues treatments are efficient both as monotherapy and in combination therapy with metformin, sulfonylureas, thiazolidinediones or insulin. Furthermore, GLP-1 receptor agonists reduce body weight, whereas DPP -4 inhibitors are body weight neutral. The treatment is safe with very low risk for adverse events, including hypoglycemia. GLP-1 based therapy is now well established therapy of type 2 diabetes, with a particular value in combination with metformin in patients who are inadequately controlled by metformin alone (Ahren B. Exp Cell Res. 2011 May 15; 317(9): 1239- 45). GLP-1 is a peptide-hormone in need of amidation by PAM.

[0192] The pharmaceutical product of this invention combining PAM and GLP-l-Gly (SEQ ID No.: 39-40 and or combining PAM, and GLP-l-Gly (SEQ ID No.: 39-40) with Vitamin C may be used for treatment and / or prevention of type-2 diabetes by taking advantage of the surprising effect of elevation of a Peptide-NH2 bioavailability when its c-terminally glycine extended biosynthetic precursor is combined with PEG-PAM and / or PEG-PAM with Vitamin C, as shown in example 5, thereby granting a prolonged elevation of amidated GLP-1 in circulation.

[0193] (5) Cardiovascular diseases prevented or treated by the pharmaceutical composition according to an embodiment of the present invention are, in particular, heart failure, myocardial infarction (e.g., acute myocardial infarction), arrhythmia (e.g., arrhythmia after catheter ablation surgery), atrial fibrillation, pulmonary hypertension or peripheral. It is a vascular disease.

[0194] The state of the art discloses the use of modified bio-ADM, for preventing or treating inflammatory bowel disease by administering said peptides into circulation and thereby elevating the concentration of said peptides in circulation. Thereby either modified or non-modified forms of ADM, or peptides with ADM activity or salts thereof are claimed. Thereby the modifications comprise i.a. replacement of the disulfide bond in ADM, but no N-terminal attachments. Thus subject matter of the present invention is also a combination of Peptide-Gly for use in treatment of IBD, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), and / or in combination with PAM and Vitamin C. Surprisingly, said combination leads to a sustained elevation of bio-ADM as shown in example 5, and others than in W02012096411A1 higher levels of active bio-ADM are reached with a reduced infusion time. Moreover, in terms of the present invention, no modification of Peptide-Gly is necessary to achieve a prolonged elevation of bioavailability of the resulting bio-ADM peptide.

[0195] An embodiment of the present invention is ADM-Gly for use in treatment of a disease in a subject, wherein ADM-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and in addition is to be used with Vitamin C.

[0196] A further embodiment of the present invention is GLP-l-Gly for use in treatment of a disease in a subject, wherein GLP-l-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and in addition is to be used with Vitamin C.

[0197] The state of the art discloses the use of PEG based prodrugs of bio-ADM, to treat or prevent diseases, such as cardiovascular, edematous and / or inflammatory diseases. The prodrug thereby should ensure a slow release of the biologically active compound.

[0198] The state of the art discloses biologically active, stabilized compound of bio-ADM, to treat or prevent diseases, such as cardiovascular, edematous and / or inflammatory diseases. The stabilization of bio- ADM is achieved by modification of bio-ADM with, but not limited to, a PEG moiety, a C3-C100 carboxylic acid moiety, an albumin moiety and others, optionally including a linker region.

[0199] In the state of the art was shown that the known effects of bio-ADM in relevant disease models may be reached using at least some of the claimed modified forms of bio-ADM. Those effects are reduction of pulmonary arterial pressure and decrease of pulmonary edema, improvement of arterial oxygenation and reduction of vascular leakage and / or hyperpermeability in vivo and in vitro. Such endpoints are relevant for assessing drug candidates effect with respect to treatment and / or prevention of cardiovascular, edematous and / or inflammatory diseases, such as sepsis or septic shock.

[0200] The subject matter of the present invention is in one embodiment a combination of Peptide-Gly for use in treatment and / or prevention of cardiovascular, edematous and / or inflammatory diseases, such as sepsis or septic shock, wherein Peptide-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM), and / or in combination with PAM and Vitamin C, wherein administered Peptide-Gly is amidated by administered PAM in vivo. Surprisingly, said combination leads to a sustained elevation of bio-ADM as shown in example 5.

[0201] The surprising and unexpected effect of said combination is the prolonged elevation of resulting bio- ADM in circulation in comparison to the short bioavailability of wz'Wty e-bio-ADM when administered alone. Other than in the state of the art, an elevation of active bio-ADM levels in circulation is reached without modifications of Peptide-Gly in said combination. Therefore, said combinations according to the present invention are suitable for treatment and / or prevention of cardiovascular, edematous and / or inflammatory diseases, in particular as sepsis or septic shock.

[0202] The state of the art discloses the use of modified bio-ADM, as a long-acting ADM derivative for prevention or treatment of i.a. cardiovascular, inflammatory or peripheral vascular diseases. Thereby ADM is N-terminally modified with a palmitoyl group or a polyethylene glycol (PEG) group, while the modification may be linked to the peptide via a linker group. In the state of the art, it was shown that the known effects of bio-ADM in relevant disease models may be reached using at least some of the claimed modified forms of bio-ADM. Those effects are reduction of pulmonary arterial pressure and decrease of pulmonary edema, improvement of arterial oxygenation and reduction of vascular leakage and / or hyperpermeability in vivo. Such endpoints are relevant for assessing drug candidates' effect with respect to treatment and / or prevention of cardiovascular, edematous and / or inflammatory diseases, in particular sepsis or septic shock.

[0203] The subject matter of the present invention is also a combination of Peptide-Gly for use in treatment of and / or prevention of cardiovascular, edematous and / or inflammatory diseases, such as sepsis or septic shock., wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), and / or in combination with PAM and Vitamin C, wherein administered Peptide-Gly is amidated by administered PAM in vivo.

[0204] Surprisingly, when ADM-Gly (SEQ ID No.: 14) is combined with PEG-PAM and / or Vitamin C, said combination leads to a sustained elevation of bio-ADM, as shown in example 5. The surprising and unexpected effect of said combination is the prolonged elevation of resulting bio-ADM in circulation in comparison to the short bioavailability of wz / ty e-bio-ADM when administered alone. Others than in the state of the art, an elevation of active bio-ADM levels in circulation is reached without modifications of ADM-Gly in said combination. Therefore, said combinations as the subject matter of the present invention are suitable for treatment and / or prevention of cardiovascular, edematous and / or inflammatory diseases, such as sepsis or septic shock.

[0205] The state of the art discloses the use of modified bio-ADM as a prophylactic or therapeutic agent for treating or preventing a neurodegenerative disease with abnormal protein accumulation. Thereby either modified or non-modified forms of ADM, or peptides with ADM activity or salts thereof are claimed. The modifications comprise, i.e. a palmitoyl group or a polyethylene glycol (PEG) group in the range of 1-100 kDa. In the state of the art the effect of claimed modified forms of bio-ADM in an Alzheimer’s model in mice was analyzed with respect to short-term memory. The analyzed modified forms of bio- ADM showed improvement of short-term memory in said model.

[0206] The subject matter of the present invention is also a combination of Peptide-Gly for use in treatment and / or or preventing a neurodegenerative disease with abnormal protein accumulation, wherein Peptide- Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), and / or in combination with PAM and Vitamin C, wherein administered Peptide-Gly is amidated by administered PAM in vivo. Surprisingly, when ADM-Gly (SEQ ID No.: 14) is combined with PEG-PAM and / or Vitamin C, said combination leads to a sustained elevation of bio-ADM as shown in example 5. The surprising and unexpected effect of said combination is the prolonged elevation of resulting bio-ADM in circulation in comparison to the short bioavailability of wz / ty e-bio-ADM when administered alone. Others than in the state of the art elevation of active bio-ADM levels in circulation is reached without modifications of ADM-Gly in said combination. Therefore, said combinations as the subject matter of the present invention are suitable for treating or preventing a neurodegenerative disease with abnormal protein accumulation.

[0207] The state of the art discloses novel long-lasting bio-ADM, derivatives and fragments and / or modifications thereof, whereas the peptide is modified with serum albumin, that may be linked to the peptide with a linker. Further the use of the described ADM derivatives for preventing or treating of several diseases, such as cardiovascular diseases, Kidney, brain and neurological diseases, genitourinary and gastrointestinal diseases, orthopedic diseases, endocrine and metabolic disorders, respiratory disorders, immune disorders and also other diseases, such as sepsis, shock autoimmune or wound healing diseases is claimed. In the state of the art, it was shown that the known effects of bio-ADM in relevant disease models may be reached using at least some of the claimed modified forms of bio-ADM. Those effects are reduction of systolic and diastolic blood-pressure and reduction of severeness of induced colitis, both in models in rats. Such endpoints are relevant for assessing drug candidates' effect with respect to treatment and / or prevention of cardiovascular, and / or inflammatory diseases of the gastrointestinal tract.

[0208] The subject matter of the present invention is also a combination of Peptide-Gly for use in treatment and / or or preventing a cardiovascular, and / or inflammatory diseases of the gastrointestinal tract, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), and / or in combination with PAM and Vitamin C, wherein administered Peptide-Gly is amidated by administered PAM in vivo. Surprisingly, when ADM-Gly (SEQ ID No.: 14) is combined with PEG- PAM and / or Vitamin C said combination leads to a sustained elevation of bio-ADM as shown in example 5. The surprising and unexpected effect of said combination is the prolonged elevation of resulting bio-ADM in circulation in comparison to the short bioavailability of wz / ty e-bio-ADM when administered alone. Others than in the state of the art elevation of active bio-ADM levels in circulation is reached without modifications of ADM-Gly in said combination. Therefore, said combinations as the subject matter of the present invention are suitable for treating or preventing a cardiovascular and / or inflammatory diseases.

[0209] The pharmaceutical composition according to the present invention is a medication that surprisingly allows for the use of non-modified Peptide-hormone-forms including a slow and long -lasting enrichment of biologically active peptides, such as ADM, from their inactive precursors, but is not limited to ADM and is therefore superior over the cited prior art. The pharmaceutical composition of the present invention is a medication that allows for more efficient treating or preventing of diseases such as Brain / neurological disorders, infectious and inflammatory diseases selected from the group comprising SIRS, sepsis, and septic shock, Gastrointestinal diseases including inflammatory diseases such as inflammatory bowel disease or Crohn's disease and ulcerative diseases, endocrine and metabolic disorders, such as diabetes and diabetic organ disorders, cardiovascular diseases such as heart failure, pulmonary hypertension, arteriosclerosis obliterans, myocardial infarction, lymphedema, Kawasaki disease, myocarditis, arrhythmia, atrial fibrillation, aortitis, pulmonary hypertension, hypertension, organ damage due to hypertension, peripheral vascular disease, and arteriosclerosis, by a controllable release of bioactive peptide hormones as shown in Example 5, such as bio-ADM, from their inactive precursors, such as ADM-Gly. As shown in Example 7, the pharmaceutical composition of the present invention, in particular comprising ADM-Gly, is surprisingly capable of treating and / or preventing endothelial dysfunction shown via the example of BBB-disruption prevention and is therefore suitable for treating and / or preventing a large number of diseases associated with endothelial- and / or blood-brain barrier dysfunctions, as, but not limited to, specified above.

[0210] The pharmaceutical composition according to the present invention is a medication that is preferably used for the prevention or treatment of symptoms, diseases, and / or disorders as described above (e.g., cardiovascular disease, brain and nerve disorders, or endocrine and gastrointestinal disorders).

[0211] In one embodiment of the present invention, the pharmaceutical composition is used to treat and / or prevent cardiovascular diseases such as heart failure, pulmonary hypertension, arteriosclerosis obliterans, Buerger's disease, myocardial infarction, lymphedema, Kawasaki disease, myocarditis, arrhythmia (for example, arrhythmia after catheter ablation surgery), atrial fibrillation, Aortitis, hypertension, organ damage due to hypertension, peripheral vascular disease, and arteriosclerosis.

[0212] In the preferred embodiment of the present invention, the pharmaceutical composition is used to treat and / or prevent heart failure, pulmonary hypertension, hypertension, organ damage due to hypertension, peripheral vascular disease, and arteriosclerosis.

[0213] In the most preferred embodiment of the present invention, the pharmaceutical composition is used to treat and / or pulmonary hypertension and hypertension.

[0214] In one embodiment of the present invention, the pharmaceutical composition is used to treat and / or prevent endocrine and metabolic disorders such as diabetes and diabetic organ disorders (e.g., diabetic nephropathy or diabetic retinopathy).

[0215] In the preferred embodiment of the present invention, the pharmaceutical composition is used to treat and / or prevent type-2 diabetes. In the specific embodiment of the present invention, the pharmaceutical composition is used to treat and / or prevent type-2 diabetes, wherein the pharmaceutical combination comprises the use of GLP-1- Gly (SEQ ID No.: 39-40) as Peptide-Gly.

[0216] In one embodiment of the present invention, the pharmaceutical composition is used to treat and / or prevent gastrointestinal diseases such as inflammatory diseases (e.g. inflammatory bowel disease or Crohn's disease), ulcerative diseases (e.g. ulcerative colitis), intestinal Behcet's disease, hepatitis, liver fibrosis, cirrhosis, and liver failure.

[0217] In the preferred embodiment of the present invention, the pharmaceutical composition is used to treat and / or prevent inflammatory diseases such as inflammatory bowel disease or Crohn's disease, ulcerative diseases such as ulcerative colitis and / or intestinal Behcet's disease.

[0218] In the most preferred embodiment of the present invention, the pharmaceutical composition is used to treat and / or prevent inflammatory bowel disease and / or Crohn's disease.

[0219] In one embodiment of the present invention, the pharmaceutical composition is used to treat and / or prevent infectious diseases caused by infectious organisms such as bacteria, viruses, fungi or parasites, more particularly by infectious bacteria, particularly diseases selected from the group comprising SIRS, sepsis, and septic shock.

[0220] In the preferred embodiment of the present invention, the pharmaceutical composition is used to treat and / or prevent sepsis and / or septic shock.

[0221] In one embodiment of the present invention, the pharmaceutical composition is used to treat and / or prevent brain and neurological diseases such as cerebral infarction, mild cognitive impairment, dementia, cerebrovascular dementia, Alzheimer's disease, and encephalitis.

[0222] In the preferred embodiment of the present invention, the pharmaceutical composition is used to treat and / or prevent cerebral infarction.

[0223] In the more preferred embodiment of the present invention, the pharmaceutical composition is used to treat and / or prevent dementias, including mild cognitive impairment, cerebrovascular dementia, and Alzheimer’s disease.

[0224] In the most preferred embodiment of the present invention, the pharmaceutical composition of the present invention is used for the prevention and / or treatment of symptoms associated with mild cognitive impairment (MCI) and Alzheimer’s Disease (AD).

[0225] In one embodiment of the present invention, the pharmaceutical composition is used to treat and / or prevent pathological disorders associated with endothelial and / or blood-brain barrier disorders.

[0226] In the preferred embodiment of the present invention, the pharmaceutical composition is used to treat and / or endothelial and / or blood-brain barrier disfunction associated with sepsis or septic shock. The person skilled in the art would readily understand that the pharmaceutical composition is used to treat and / or prevent endothelial and / or blood-brain barrier disfunction associated with sepsis or septic shock.

[0227] In the more preferred embodiment of the present invention, the pharmaceutical composition is used to treat and / or prevent endothelial and / or blood-brain barrier dysfunction associated with cerebral infarction.

[0228] In the more preferred embodiment of the present invention, the pharmaceutical composition is used to treat and / or prevent endothelial and / or blood-brain barrier dysfunction associated with dementias, including mild cognitive impairment, cerebrovascular dementia, and Alzheimer’s disease.

[0229] In the most preferred embodiment of the present invention, the pharmaceutical composition is used to treat and / or prevent endothelial and / or blood-brain barrier dysfunction associated with mild cognitive impairment (MCI) and Alzheimer’s Disease (AD).

[0230] More preferably, it is used for the prevention or treatment of heart failure, acute myocardial infarction, arrhythmia, atrial fibrillation, pulmonary hypertension, peripheral vascular disease, stroke, dementia, inflammatory bowel disease, Crohn's disease, ulcerative colitis, intestinal Behcet's disease, diabetes, diabetic nephropathy, diabetic retinopathy, pulmonary fibrosis, sepsis, or septic shock. By using the drug of this embodiment for the prevention or treatment of the symptoms, diseases and / or disorders described above, it is possible to exhibit superior pharmacokinetics compared to naturally occurring Peptide-NH2, and the preventive or therapeutic effect is substantially equivalent or superior to that of naturally occurring Peptide-NH2.

[0231] As used herein, "prevention" means substantially preventing the occurrence (onset or manifestation) of symptoms, diseases and / or disorders. Also, as used herein, "treatment" means suppressing (e.g., suppressing progression), alleviating, repairing and / or curing symptoms, diseases and / or disorders that have occurred (onset or manifestation).

[0232] Subject matter according to the invention is Peptide-Gly for use in treatment of a disease in a subject, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM, SEQ ID No.: 1-10) and in addition to be used with Vitamin C.

[0233] An embodiment of the present invention is Peptide-Gly for use in treatment of a disease in a subject, wherein Peptide-Gly is ADM-Gly and wherein ADM-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM, SEQ ID No.: 1-10) and in addition to be used with Vitamin C.

[0234] An embodiment of the present invention is Peptide-Gly for use in treatment of a disease in a subject, wherein Peptide-Gly is GLP-l-Gly and wherein GLP-l-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM, SEQ ID No.: 1-10) and in addition to be used with Vitamin C.

[0235] Subject matter according to the invention is Peptide-Gly for use in treatment of a disease in a subject, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM, SEQ ID No.: 1-10) and wherein Peptide-Gly and PAM are to be administered consecutively, or Peptide-Gly and PAM are to be administered simultaneously.

[0236] An embodiment of the invention is ADM-Gly for use in treatment of a disease in a subject, wherein ADM-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM, SEQ ID No.: 1-10) and wherein ADM-Gly and PAM are to be administered consecutively, or ADM-Gly and PAM are to be administered simultaneously.

[0237] Another embodiment of the invention is GLP-l-Gly for use in treatment of a disease in a subject, wherein GLP-l-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM, SEQ ID No.: 1-10) and wherein GLP-l-Gly and PAM are to be administered consecutively, or GLP-l-Gly and PAM are to be administered simultaneously.

[0238] Other embodiments of the invention relate to Peptide-Gly for use in treatment of a disease in a subject, wherein Peptide-Gly is to be used in combination with recombinant PAM (SEQ ID No.: 47) and in addition to be used with Vitamin C.

[0239] Further embodiments of the present invention relate to Peptide-Gly for use in treatment of a disease in a subject, wherein Peptide-Gly is ADM-Gly and wherein ADM-Gly is to be used in combination with recombinant PAM (SEQ ID No.: 47) and in addition to be used with Vitamin C.

[0240] Other embodiments of the present invention relate to Peptide-Gly for use in treatment of a disease in a subject, wherein Peptide-Gly is GLP-l-Gly and wherein GLP-l-Gly is to be used in combination with recombinant PAM (SEQ ID No.: 47) and in addition to be used with Vitamin C.

[0241] Certain embodiments of the invention relate to Peptide-Gly for use in treatment of a disease in a subject, wherein Peptide-Gly is to be used in combination with recombinant PAM (SEQ ID No.: 47) and wherein Peptide-Gly and PAM are to be administered consecutively, or Peptide-Gly and PAM are to be administered simultaneously.

[0242] Further embodiments of the invention relate to ADM-Gly for use in treatment of a disease in a subject, wherein ADM-Gly is to be used in combination with recombinant PAM (SEQ ID No.: 47) and wherein ADM-Gly and PAM are to be administered consecutively, or ADM-Gly and PAM are to be administered simultaneously.

[0243] Other embodiments of the invention relate to GLP-l-Gly for use in treatment of a disease in a subject, wherein GLP-l-Gly is to be used in combination with recombinant PAM (SEQ ID No.: 47) and wherein GLP-l-Gly and PAM are to be administered consecutively, or GLP-l-Gly and PAM are to be administered simultaneously.

[0244] The threshold according to the present invention may be predetermined by measuring, such as the level of PAM and / or its isoforms and / or fragments in a samples obtained from a cohort of subjects and calculating e.g. the 25th percentile, more particularly the 10th percentile, even more particularly the 5th percentile to define the threshold by which subjects are characterized as suffering from a medical condition or as being at risk of getting a medical condition.

[0245] In the following examples for thresholds according to the present invention are provided, however, the person skilled in the art is aware on how to determine a specific threshold.

[0246] Subject matter according to the invention is Peptide-Gly for use in treatment of a disease in a subject, wherein the molar ratio of said Peptide-Gly and its corresponding Peptide-amide (Peptide-NEE) (Peptide-Gly / Peptide-amide) in a bodily fluid is elevated over 1.5, preferably elevated over 1.6, more preferably elevated over 1.7, more preferably is elevated over 1.8, more preferable is elevated over 1.9, more preferably is higher than 2 and / or the concentration of PAM (SEQ ID No. 1-10) in a bodily fluid is equal or below 100 ng / mL (100 ng / mL to 0 ng / mL), preferably equal or below 90 ng / mL (90 ng / mL to 0 ng / mL), more preferably equal or below 80 ng / mL (80 ng / mL to 0 ng / mL), more preferably equal or below 60 ng / mL (60 ng / mL to 0 ng / mL), more preferably equal or below 50 ng / mL (50 ng / mL to 0 ng / mL) and most preferably equal or below 40 ng / mL (40 ng / mL to 0 ng / mL) and / or the activity of PAM (SEQ ID No. 1-10) in a bodily fluid is equal or below 24 Units / L, more preferably equal or below 20 Units / L, more preferably equal or below 16 Units / L, more preferably equal or below 14 Units / L and most preferably equal or below 10 Units / L, wherein the quantification of activity of PAM in Units is described in Example 2 and the term Units / L defines the activity of PAM in Units per 1 L of sample material and / or the Peptide-NH2 form of said Peptide-Gly is equal or below 20 pg / mL (20 pg / mL-0 pg / mL), preferably equal or below 15 pg / mL (15 pg / mL - 0 pg / mL), preferably equal or below 10 pg / mL (10 pg / mL-0 pg / mL), preferably equal or below 5 pg / mL (5 pg / mL - 0 pg / mL) and / or said Peptide-Gly is above 15 pg / mL, preferably equal or above 20 pg / mL, preferably equal or above 30 pg / mL, preferably is equal or above 50 pg / mL, wherein given levels of said Peptide-Gly may refer to a range of 15 - 10000 pg / mL, preferably to 20 - 8000 pg / mL, more preferably to 30- 5000 pg / mL, and most preferably in the range of 50-4500 pg / mL and / or the Peptide-NH2 form of said Peptide-Gly is equal or above 70 pg / mL (70-3000 pg / mL), irrespective of the status of PAM and / or Peptide-Gly, and / or the concentration of PAM (SEQ ID No.: 1-10) is equal or above 80 ng / mL (80-1000 ng / mL), preferably equal or above 100 ng / mL (100-1000 ng / mL), irrespective of the said Peptide-Gly and the corresponding Peptide-NH2 status, and / or the activity of PAM (SEQ ID No.: 1-10) is equal or above 14 Units / L (14-130 Units / L), preferably equal or above 20 Units / L (20-120 Units / L), preferably equal or above 24 units / L (24-115 Units / L), irrespective of the said Peptide-Gly and the corresponding Peptide-NEEstatus, wherein the quantification of activity of PAM in Units is described in Example 2 and the term Units / L defines the activity of PAM in Units per 1 L of sample material.

[0247] An embodiment of the present invention is ADM-Gly (SEQ ID No.: 14, 17 - 21, 27) for use in treatment of a disease in a subject, wherein the molar ratio of ADM-Gly (SEQ ID No.: 14, 17 - 21, 27) and bio- ADM (SEQ ID No.: 15, 22-26, 28) (ADM-Gly / bio-ADM) in a bodily fluid is elevated over 1.5, preferably elevated over 1.6, more preferably elevated over 1.7, more preferably is elevated over 1.8, more preferable is elevated over 1.9, more preferably is higher than 2, and / or the concentration of PAM (SEQ ID No. 1-10) in a bodily fluid is equal or below 100 ng / mL (100 ng / mL to 0 ng / mL), preferably equal or below 90 ng / mL (90 ng / mL to 0 ng / mL), more preferably equal or below 80 ng / mL (80 ng / mL to 0 ng / mL), more preferably equal or below 60 ng / mL (60 ng / mL to 0 ng / mL), more preferably equal or below 50 ng / mL (50 ng / mL to 0 ng / mL) and most preferably equal or below 40 ng / mL (40 ng / mL to 0 ng / mL) and / or the activity of PAM (SEQ ID No. 1-10) in a bodily fluid is below 24 Units / L, more preferably below 20 Units / L, more preferably below 16 Units / L, more preferably below 14 Units / L and most preferably below 10 Units / L, wherein the quantification of activity of PAM in Units is described in Example 2 and the term Units / L defines the activity of PAM in Units per 1 L of sample material and / or bio-ADM (SEQ ID No.: 15, 22-26, 28) is equal or below 20 pg / mL (20 pg / mL-0 pg / mL), preferably equal or below 15 pg / mL (15 pg / mL - 0 pg / mL), preferably equal or below 10 pg / mL (10 pg / mL-0 pg / mL), preferably equal or below 5 pg / mL (5 pg / mL - 0 pg / mL) and / or

[0248] ADM-Gly (SEQ ID No.: 14, 17-21, 27) is equal or above 15 pg / mL, preferably equal or above 20 pg / mL, preferably equal or above 30 pg / mL, preferably is equal or above 50 pg / mL, wherein given levels of ADM-Gly may refer to a range of 15 - 10000 pg / mL, preferably to 20 - 8000 pg / mL, more preferably to 30- 5000 pg / mL, and most preferably in the range of 50-4500 pg / mL and / or bio-ADM (SEQ ID No.: 15, 22-26, 28) is equal or above 70 pg / mL (70-3000 pg / mL), irrespective of the status of PAM and / or ADM-Gly, and / or the concentration of PAM (SEQ ID No.: 1-10) is equal or above 80 ng / mL (80-1000 ng / mL), preferably equal or above 100 ng / mL (100-1000 ng / mL), irrespective of the bio-ADM (SEQ ID No.: 15, 22-26, 28) and ADM-Gly (SEQ ID No.: 14, 17-21, 27) status, and / or the activity of PAM (SEQ ID No.: 1-10) is equal or above 14 Units / L (14-130 Units / L), preferably equal or above 20 Units / L (20-120 Units / L), preferably equal or above 24 units / L (24-115 Units / L), irrespective of bio-ADM (SEQ ID No.: 15, 22-26, 28) and ADM-Gly (SEQ ID No.: 14, 17-21, 27) status, wherein the quantification of activity of PAM in Units is described in Example 2 and the term Units / L defines the activity of PAM in Units per 1 L of sample material.

[0249] Another embodiment of the present invention is GLP-l-Gly (SEQ ID No.: 39-40) for use in treatment of a disease in a subject, wherein the molar ratio of GLP-l-Gly (SEQ ID No.: 39-40) and GLP-I-NH2 (GLP-l-Gly / GLP-I-NH2) in a bodily fluid is elevated over 1.5, preferably elevated over 1.6, more preferably elevated over 1.7, more preferably is elevated over 1.8, more preferable is elevated over 1.9, more preferably is higher than 2, and / or the concentration of PAM (SEQ ID No. 1-10) in a bodily fluid is equal or below 100 ng / mL (100 ng / mL to 0 ng / mL), preferably equal or below 90 ng / mL (90 ng / mL to 0 ng / mL), more preferably equal or below 80 ng / mL (80 ng / mL to 0 ng / mL), more preferably equal or below 60 ng / mL (60 ng / mL to 0 ng / mL), more preferably equal or below 50 ng / mL (50 ng / mL to 0 ng / mL) and most preferably equal or below 40 ng / mL (40 ng / mL to 0 ng / mL) and / or the activity of PAM (SEQ ID No. 1-10) in a bodily fluid is equal or below 24 Units / L, more preferably equal or below 20 Units / L, more preferably equal or below 16 Units / L, more preferably equal or below 14 Units / L and most preferably equal or below 10 Units / L, wherein the quantification of activity of PAM in Units is described in Example 2 and the term Units / L defines the activity of PAM in Units per 1 E of sample material and / or

[0250] GLP-I-NH2 is equal or below 20 pg / mL (20 pg / mL-0 pg / mL), preferably equal or below 15 pg / mL (15 pg / mL - 0 pg / mL), preferably equal or below 10 pg / mL (10 pg / mL-0 pg / mL), preferably equal or below 5 pg / mL (5 pg / mL - 0 pg / mL) and / or

[0251] GLP-l-Gly (SEQ ID No.: 39-40) is equal or above 15 pg / mL, preferably equal or above 20 pg / mL, preferably equal or above 30 pg / mL, preferably is equal or above 50 pg / mL, wherein given levels of GLP-l-Gly (SEQ ID No.: 39-40) may refer to a range of 15 - 10000 pg / mL, preferably to 20 - 8000 pg / mL, more preferably to 30- 5000 pg / mL, and most preferably in the range of 50-4500 pg / mL and / or GLP-I-NH2 is equal or above 70 pg / mL (70-3000 pg / mL), irrespective of the status of PAM and / or GLP-l-Gly (SEQ ID NO.: 39-40), and / or the concentration of PAM (SEQ ID No.: 1-10) is equal or above 80 ng / mL (80-1000 ng / mL), preferably equal or above 100 ng / mL (100-1000 ng / mL), irrespective of the GLP-I-NH2 and GLP-l-Gly (SEQ ID No.: 39-40) status, and / or the activity of PAM (SEQ ID No.: 1-10) is equal or above 14 Units / L (14-130 Units / L), preferably equal or above 20 Units / L (20-120 Units / L), preferably equal or above 24 units / L (24-115 Units / L), irrespective of GLP-I-NH2 and GLP-l-Gly (SEQ ID No.: 39-40) status, wherein the quantification of activity of PAM in Units is described in Example 2 and the term Units / L defines the activity of PAM in Units per 1 L of sample material.

[0252] In one embodiment, said molar ratio of said Peptide-Gly and its corresponding Peptide-amide (Pcptidc-NFf) (Peptide- Gly / Peptide-amide) and / or concentration of Pcptidc-NFf and / or concentration of Peptide-Gly and / or concentration of PAM and / or activity of PAM in a bodily fluid of the subject may be determined at least once before and / or at least once after the treatment of the subject.

[0253] In a more specific embodiment, said

[0254] - molar ratio of ADM-Gly (SEQ ID No.: 14, 17 - 21, 27) and bio-ADM (SEQ ID No.: 15, 22- 26, 28) (ADM-Gly / bio-ADM) and / or concentration of bio-ADM (SEQ ID No.: 15, 22-26, 28) and / or concentration of ADM-Gly (SEQ ID No.: 14, 17 - 21, 27) and / or concentration of PAM and / or activity of PAM in a bodily fluid of the subject may be determined at least once before and / or at least once after the treatment of the subject.

[0255] In another more specific embodiment, said

[0256] - molar ratio of GLP-l-Gly (SEQ ID No.: 39-40) and GLP-1-NH2(GLP-l-Gly / GLP-1-NH2) and / or concentration of GLP- 1 -NH2and / or concentration of GLP-l-Gly (SEQ ID No.: 39-40) and / or concentration of PAM and / or activity of PAM in a bodily fluid of the subject may be determined at least once before and / or at least once after the treatment of the subject.

[0257] A person skilled in the art will understand, that the afore mentioned concentration of PAM or activity of PAM comprises endogenous PAM and / or its isoforms and / or fragments thereof, as well as exogenous (therapeutically applied) PAM and / or its isoforms and / or fragments thereof including recombinant PAM SEQ ID No.: 47 either modified or unmodified, after said subject has been treated with said PAM.

[0258] Subject matter of the present invention is Peptide-Gly for use in treatment of a disease in a subject, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein Peptide-Gly is applied at a dosage of 0. 1-500 pg / kg, preferably 1-400 pg / kg, more preferably 2-300 pg / mL, more preferably 3-200 pg / kg, more preferably 5-100 pg / kg and wherein PAM is applied at a dosage of 1.5 - 54 Units / kg, preferably 1.7-33 Units / kg, more preferably 1.8-26 Units / kg, more preferably 1.9-20.6 Units / kg, more preferably 2-14 Units / kg, wherein preferably combined as 0.1- 500 pg / kg of Peptide-Gly with 1.5-54 Units / kg of PAM, more preferably 1-400 pg / kg of Peptide-Gly with 1.7-33 Units / kg PAM, more preferably 2-300 pg / kg Peptide-Gly with 1.8-26 Units / kg PAM, more preferably 3-200 pg / kg of Peptide-Gly with 1.9-20.6 Units / kg of PAM, more preferably 5-100 pg / kg of Peptide-Gly with 2-14 Units / kg of PAM.

[0259] The skilled person readily recognizes that typically, pharmaceutical agents are administered to a subject in the unit weight / bodyweight of the subject. Subject matter of the present invention is therefore Peptide- Gly for use in treatment of a disease in a subject, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein Peptide-Gly is applied at a dosage of 0.1-500 jj.g / kg, preferably 1-400 pg / kg, more preferably 2-300 pg / kg, more preferably 3-200 pg / kg, more preferably 5-100 pg / kg and wherein PAM is applied at a dosage of 1.5 - 54 Units / kg, preferably 1.7-33 Units / kg, more preferably 1.8-26 Units / kg, more preferably 1.9-20.6 Units / kg, more preferably 2-14 Units / kg, wherein preferably combined as 0.1-500 pg / kg of Peptide-Gly with 1.5-54 Units / kg of PAM, more preferably 1-400 pg / kg of Peptide-Gly with 1.7-33 Units / kg PAM, more preferably 2-300 pg / kg Peptide-Gly with 1.8-26 Units / kg PAM, more preferably 3-200 pg / kg of Peptide-Gly with 1.9- 20.6 Units / kg of PAM, more preferably 5-100 pg / kg of Peptide-Gly with 2-14 Units / kg of PAM.

[0260] Another embodiment of the present invention is ADM-Gly for use in treatment of a disease in a subject, wherein ADM-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein ADM-Gly is applied at a dosage of 0.1-500 pg / kg, preferably 1-400 pg / kg, more preferably 2-300 pg / mU, more preferably 2-300 pg / kg, more preferably 3-200 pg / kg, more preferably 5-100 pg / kg and wherein PAM is applied at a dosage of 1.5 - 54 Units / kg, preferably 1.7-33 Units / kg, more preferably 1.8-26 Units / kg, more preferably 1.9-20.6 Units / kg, more preferably 2-14 Units / kg, wherein preferably combined as 0.1-500 pg / kg of ADM-Gly with 1.5-54 Units / kg of PAM, more preferably 1-400 pg / kg of ADM-Gly with 1.7-33 Units / kg PAM, more preferably 2-300 pg / kg ADM- Gly with 1.8-26 Units / kg PAM, more preferably 3-200 pg / kg of ADM-Gly with 1.9-20.6 Units / kg of PAM, more preferably 5-100 pg / kg of GUP-l-Gly with 2-14 Units / kg of PAM.

[0261] A further embodiment of the present invention is Peptide-Gly for use in treatment of a disease in a subject, wherein GUP-l-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein GUP-l-Gly is applied at a dosage of 0.1-500 pg / kg, preferably 1- 400 pg / kg, more preferably 2-300 pg / mU, more preferably 2-300 pg / kg, more preferably 3-200 pg / kg, more preferably 5-100 pg / kg and wherein PAM is applied at a dosage of 1.5 - 54 Units / kg, preferably 1.7-33 Units / kg, more preferably 1.8-26 Units / kg, more preferably 1.9-20.6 Units / kg, more preferably 2-14 Units / kg, wherein preferably combined as 0.1-500 pg / kg of GUP-l-Gly with 1.5-54 Units / kg of PAM, more preferably 1-400 pg / kg of GUP-l-Gly with 1.7-33 Units / kg PAM, more preferably 2-300 pg / kg GUP-l-Gly with 1.8-26 Units / kg PAM, more preferably 3-200 pg / kg of GUP-l-Gly with 1.9-20.6 Units / kg of PAM, more preferably 5-100 pg / kg of GUP-l-Gly with 2-14 Units / kg of PAM.

[0262] In particular embodiments of the present invention, said pharmaceutical composition is formulated to be administered or is administered orally, epicutaneously, subcutaneously, intradermally, sublingually, intramuscularly, intraarterially, intravenously, via the central nervous system (CNS, intracerebrally, intracerebroventricularly, intrathecally) or via intraperitoneal administration, particularly epicutaneously, subcutaneously, intradermally, intramuscularly, or intraperitoneally, more particularly subcutaneously, intramuscularly, or intraperitoneally; “formulated to be administered (e.g. orally)” likewise reads on “formulated for (e.g. oral) administration”. In preferred embodiments of the present invention, said pharmaceutical composition is formulated to be administered or is administered, epicutaneously, subcutaneously, intradermally, intramuscularly, intraarterially, intravenously, via the central nervous system (CNS, intracerebrally, intracerebroventricularly, intrathecally) or via intraperitoneal administration, particularly epicutaneously, subcutaneously, intradermally, intramuscularly, or intraperitoneally, more particularly subcutaneously, intramuscularly, or intraperitoneally; “formulated to be administered (e.g. orally)” likewise reads on “formulated for (e.g. oral) administration.”

[0263] In the most preferred embodiments of the present invention, said pharmaceutical composition is formulated to be administered or is administered, subcutaneously, intradermally, intramuscularly, intraarterially or intravenously, or via intraperitoneal administration.

[0264] According to the present invention the applied dosage might be a single bolus injection delivering the described amount of the compounds to be applied or a continuous infusion of the compounds delivering the desired amount of compounds over a distinct period of time taking into account the velocity of infusion. Thereby the compounds might be applied as a combined injection and / or infusion or as several parallel injections and / or infusion, wherein one or all compounds are delivered as an injection and / or infusions or a distinct compound is injected and another distinct compound is infused. This includes for example that peptide-Gly is applied as separate injection and / or infusion and PAM is applied as separate injection and / or infusion.

[0265] Moreover, peptide-Gly and PAM, if administered separately, may be administered via the same or different routes of application. This means for example that peptide-Gly is administered subcutaneously and PAM intravenously, respectively.

[0266] It is to be understood that both, peptide-Gly and PAM are administered as exogenous substances.

[0267] Subject matter of the present invention is Peptide-Gly for use in treatment of a disease in a subject, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and in addition to be used with Vitamin C and wherein Vitamin C is applied at a dosage of 1- 10000 mg / kg, preferably 2-8000 mg / kg, more preferably 3-6000 mg / kg, more preferably 4-4000 mg / kg, more preferably 5-2000 mg / kg, more preferably 10-1000 mg / kg.

[0268] A further embodiment of the present invention is ADM-Gly for use in treatment of a disease in a subject, wherein ADM-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and in addition to be used with Vitamin C and wherein Vitamin C is applied at a dosage of 1- 10000 mg / kg, preferably 2-8000 mg / kg, more preferably 3-6000 mg / kg, more preferably 4-4000 mg / kg, more preferably 5-2000 mg / kg, more preferably 10-1000 mg / kg. Another embodiment of the present invention is GLP- 1 -Gly for use in treatment of a disease in a subject, wherein GLP- 1 -Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and in addition to be used with Vitamin C and wherein Vitamin C is applied at a dosage of 1- 10000 mg / kg, preferably 2-8000 mg / kg, more preferably 3-6000 mg / kg, more preferably 4-4000 mg / kg, more preferably 5-2000 mg / kg, more preferably 10-1000 mg / kg.

[0269] An embodiment of the present invention is Peptide-Gly for use in treatment of a disease in a subject, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and in addition to be used with Vitamin C and wherein

[0270] • preferably 0.1-500 pg / kg of Peptide-Gly and 1.5-54 Units / kg of PAM are combined with 1- 10000 mg / kg, preferably 2-8000 mg / kg, more preferably 3-6000 mg / kg, more preferably 4-4000 mg / kg, more preferably 5-2000 mg / kg, more preferably 10-1000 mg / kg of Vitamin C,

[0271] • more preferably 1-400 pg / kg of Peptide-Gly and 1.7-33 Units / kg PAM are combined with 1- 10000 mg / kg, preferably 2-8000 mg / kg, more preferably 3-6000 mg / kg, more preferably 4-4000 mg / kg, more preferably 5-2000 mg / kg, more preferably 10-1000 mg / kg of Vitamin C,

[0272] • more preferably 2-300 pg / kg of Peptide-Gly and 1.8-26 Units / kg PAM are combined with 1- 10000 mg / kg, preferably 2-8000 mg / kg, more preferably 3-6000 mg / kg, more preferably 4-4000 mg / kg, more preferably 5-2000 mg / kg, more preferably 10-1000 mg / kg of Vitamin C,

[0273] • more preferably 3-200 pg / kg Peptide-Gly and 1.9-20.6 Units / kg PAM are combined with 1- 10000 mg / kg, preferably 2-8000 mg / kg, more preferably 3-6000 mg / kg, more preferably 4-4000 mg / kg, more preferably 5-2000 mg / kg, more preferably 10-1000 mg / kg of Vitamin C,

[0274] • more preferably 5-100 pg / kg of Peptide-Gly with 2-14 Units / kg of PAM are combined with 1- 10000 mg / kg, preferably 2-8000 mg / kg, more preferably 3-6000 mg / kg, more preferably 4-4000 mg / kg, more preferably 5-2000 mg / kg, more preferably 10-1000 mg / kg of Vitamin C, wherein the applied dosage might be a single bolus injection delivering the described amount of the compounds to be applied or a continuous infusion of the compounds delivering the desired amount of compounds over a distinct period of time taking into account the velocity of infusion. Thereby the compounds might be applied as a combined injection and / or infusion or as several parallel injections and / or infusions, wherein one or all compounds are delivered as an injection and / or infusion or a distinct compound is injected and another distinct compound is infused.

[0275] Said compounds or combinations thereof may be further administered using intramuscular or intraperitoneal or subcutaneous or intravenous application routes. An embodiment of the present invention is ADM-Gly for use in treatment of a disease in a subject, wherein ADM-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and in addition to be used with Vitamin C and wherein Vitamin C is applied at a dosage of

[0276] • preferably 0.1-500 pg / kg of ADM-Gly and 1.5-54 Units / kg of PAM are combined with 1-10000 mg / kg, preferably 2-8000 mg / kg, more preferably 3-6000 mg / kg, more preferably 4-4000 mg / kg, more preferably 5-2000 mg / kg, more preferably 10-1000 mg / kg of Vitamin C,

[0277] • more preferably 1-400 pg / kg of ADM-Gly and 1.7-33 Units / kg PAM are combined with 1-10000 mg / kg, preferably 2-8000 mg / kg, more preferably 3-6000 mg / kg, more preferably 4-4000 mg / kg, more preferably 5-2000 mg / kg, more preferably 10-1000 mg / kg of Vitamin C,

[0278] • more preferably 2-300 pg / kg of ADM-Gly and 1.8-26 Units / kg PAM are combined with 1-10000 mg / kg, preferably 2-8000 mg / kg, more preferably 3-6000 mg / kg, more preferably 4-4000 mg / kg, more preferably 5-2000 mg / kg, more preferably 10-1000 mg / kg of Vitamin C,

[0279] • more preferably 3-200 pg / kg ADM-Gly and 1.9-20.6 Units / kg PAM are combined with 1-10000 mg / kg, preferably 2-8000 mg / kg, more preferably 3-6000 mg / kg, more preferably 4-4000 mg / kg, more preferably 5-2000 mg / kg, more preferably 10-1000 mg / kg of Vitamin C, more preferably 5-100 pg / kg of ADM-Gly with 2-14 Units / kg of PAM are combined with 1-10000 mg / kg, preferably 2-8000 mg / kg, more preferably 3-6000 mg / kg, more preferably 4-4000 mg / kg, more preferably 5-2000 mg / kg, more preferably 10-1000 mg / kg of Vitamin C.

[0280] A further embodiment of the present invention is GUP-l-Gly for use in treatment of a disease in a subject, wherein GUP-l-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and in addition to be used with Vitamin C and wherein Vitamin C is applied at a dosage of

[0281] • preferably 0.1-500 pg / kg of GUP-l-Gly and 1.5-54 Units / kg of PAM are combined with 1-10000 mg / kg, preferably 2-8000 mg / kg, more preferably 3-6000 mg / kg, more preferably 4-4000 mg / kg, more preferably 5-2000 mg / kg, more preferably 10-1000 mg / kg of Vitamin C,

[0282] • more preferably 1-400 pg / kg of GUP-l-Gly and 1.7-33 Units / kg PAM are combined with 1- 10000 mg / kg, preferably 2-8000 mg / kg, more preferably 3-6000 mg / kg, more preferably 4-4000 mg / kg, more preferably 5-2000 mg / kg, more preferably 10-1000 mg / kg of Vitamin C,

[0283] • more preferably 2-300 pg / kg of GUP-l-Gly and 1.8-26 Units / kg PAM are combined with 1- 10000 mg / kg, preferably 2-8000 mg / kg, more preferably 3-6000 mg / kg, more preferably 4-4000 mg / kg, more preferably 5-2000 mg / kg, more preferably 10-1000 mg / kg of Vitamin C,

[0284] • more preferably 3-200 pg / kg GUP-l-Gly and 1.9-20.6 Units / kg PAM are combined with 1- 10000 mg / kg, preferably 2-8000 mg / kg, more preferably 3-6000 mg / kg, more preferably 4-4000 mg / kg, more preferably 5-2000 mg / kg, more preferably 10-1000 mg / kg of Vitamin C, more preferably 5-100 pg / kg of GLP-l-Gly with 2-14 Units / kg of PAM are combined with 1-10000 mg / kg, preferably 2-8000 mg / kg, more preferably 3-6000 mg / kg, more preferably 4-4000 mg / kg, more preferably 5-2000 mg / kg, more preferably 10-1000 mg / kg of Vitamin C.

[0285] Subject matter of the present invention is Peptide-Gly for use in treatment of a disease in a subject, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) wherein said Peptide-Gly is modified via amino acid manipulations, preferably via fusion to Albumins, e.g. serum Albumin or recombinant serum Albumin, more preferably via non covalent binding to serum Albumin due to a conjugated fatty acid chain to PAM, mor preferably via fusion with IgG Fc regions or Transferrin, more preferably modified to act as PEG-based carrier prodrug, most preferably via post-translational modifications attaching natural or synthetic polymers, whereas the natural or synthetic polymer to be used is HAP, preferably ELP, more preferably PAS, more preferably PSA, more preferably GLK, more preferably XTEN and most preferably PEG.

[0286] An embodiment of the present invention is ADM-Gly for use in treatment of a disease in a subject, wherein ADM-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) wherein said ADM-Gly is modified by via amino acid manipulations, preferably via fusion to Albumins, e.g. serum Albumin or recombinant serum Albumin, more preferably via non covalent binding to serum Albumin due to a conjugated fatty acid chain to PAM, mor preferably via fusion with IgG Fc regions or Transferrin, more preferably modified to act as PEG-based carrier prodrug, most preferably via post-translational modifications attaching natural or synthetic polymers, whereas the natural or synthetic polymer to be used is HAP, preferably ELP, more preferably PAS, more preferably PSA, more preferably GLK, more preferably XTEN and most preferably PEG.

[0287] A further embodiment of the present invention is GLP-l-Gly for use in treatment of a disease in a subject, wherein GLP-l-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) wherein said GLP-l-Gly is modified by via amino acid manipulations, preferably via fusion to Albumins, e.g. serum Albumin or recombinant serum Albumin, more preferably via non covalent binding to serum Albumin due to a conjugated fatty acid chain to PAM, mor preferably via fusion with IgG Fc regions or Transferrin, more preferably modified to act as PEG-based carrier prodrug, most preferably via post-translational modifications attaching natural or synthetic polymers, whereas the natural or synthetic polymer to be used is HAP, preferably ELP, more preferably PAS, more preferably PSA, more preferably GLK, more preferably XTEN and most preferably PEG.

[0288] Subject matter of the present invention is Peptide-Gly for use in treatment of a disease in a subject, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) wherein said PAM is modified by pegylation, fusion with long-lasting proteins, site-directed mutagenesis, glycosylation, conjugation with lipids.

[0289] Subject matter of the present invention is Peptide-Gly for use in treatment of a disease in a subject, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) wherein said PAM is modified by pegylation.

[0290] A further embodiment of the present invention is ADM-Gly for use in treatment of a disease in a subject, wherein ADM-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) wherein said PAM is modified by pegylation.

[0291] Another embodiment of the present invention is GLP- 1 -Gly for use in treatment of a disease in a subject, wherein GLP- 1 -Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) wherein said PAM is modified by pegylation.

[0292] Subject matter of the present invention is a pharmaceutical combination or pharmaceutical composition comprising Peptide-Gly and peptidylglycine a-amidating monooxygenase (PAM), wherein said combination may be a kits of part combination with at least two pharmaceutical compositions, wherein one of which comprises Peptide-Gly as defined above and wherein the other comprises PAM as defined above or wherein said pharmaceutical composition comprises both Peptide-Gly and peptidylglycine a- amidating monooxygenase (PAM), and wherein PAM and / or Peptide-Gly may be in modified or unmodified form.

[0293] An embodiment of the present invention is a pharmaceutical combination or pharmaceutical composition comprising ADM-Gly and peptidylglycine a-amidating monooxygenase (PAM), wherein said combination may be a kits of part combination with at least two pharmaceutical compositions, wherein one of which comprises ADM-Gly as defined above and wherein the other comprises PAM as defined above or wherein said pharmaceutical composition comprises both ADM-Gly and peptidylglycine a-amidating monooxygenase (PAM), and wherein PAM and / or ADM-Gly may be in modified or unmodified form.

[0294] Another embodiment of the present invention is a pharmaceutical combination or pharmaceutical composition comprising GLP- 1 -Gly and peptidylglycine a-amidating monooxygenase (PAM), wherein said combination may be a kits of part combination with at least two pharmaceutical compositions, wherein one of which comprises GLP- 1 -Gly as defined above and wherein the other comprises PAM as defined above or wherein said pharmaceutical composition comprises both GLP- 1 -Gly and peptidylglycine a-amidating monooxygenase (PAM), and wherein PAM and / or GLP-l-Gly may be in modified or unmodified form.

[0295] In one embodiment of the invention said combination or composition comprises in addition Vitamin C.

[0296] In one embodiment of the invention of said pharmaceutical combination or pharmaceutical composition the dosage of Peptide-Gly is 0.1-500 pg / kg, preferably 1-400 pg / kg, more preferably 2-300 pg / mL, more preferably 2-300 pg / kg, more preferably 3-200 pg / kg, more preferably 5-100 pg / kg and wherein PAM dosage is in between of 1.5 - 54 Units / kg, preferably 1.7-33 Units / kg, more preferably 1.8-26 Units / kg, more preferably 1.9-20.6 Units / kg, more preferably 2-14 Units / kg.

[0297] In one embodiment 0.1-500 pg / kg of Peptide-Gly are combined with 1.5-54 Units / kg of PAM, more preferably 1-400 pg / kg of Peptide-Gly are combined with 1.7-33 Units / kg PAM, more preferably 2-300 pg / kg Peptide-Gly are combined with 1.8-26 Units / kg PAM, more preferably 3-200 pg / kg of Peptide- Gly are combined with 1.9-20.6 Units / kg of PAM.

[0298] In one embodiment 0.1-500 pg / kg of ADM-Gly are combined with 1.5-54 Units / kg of PAM, more preferably 1-400 pg / kg of ADM-Gly are combined with 1.7-33 Units / kg PAM, more preferably 2-300 pg / kg ADM-Gly are combined with 1.8-26 Units / kg PAM, more preferably 3-200 pg / kg of ADM-Gly are combined with 1.9-20.6 Units / kg of PAM.

[0299] In one embodiment 0.1-500 pg / kg of GUP-l-Gly are combined with 1.5-54 Units / kg of PAM, more preferably 1-400 pg / kg of GUP-l-Gly are combined with 1.7-33 Units / kg PAM, more preferably 2-300 pg / kg GUP-l-Gly are combined with 1.8-26 Units / kg PAM, more preferably 3-200 pg / kg of GUP-l- Gly are combined with 1.9-20.6 Units / kg of PAM.

[0300] In a preferred embodiment 5-100 pg / kg of Peptide-Gly are combined with 2-14 Units / kg of PAM.

[0301] The applied dosage might be a single bolus injection delivering the described amount of the compounds to be applied or a continuous infusion of the compounds delivering the desired amount of compounds over a distinct period of time taking into account the velocity of infusion. Thereby the compounds might be applied as a combined injection and / or infusion or as several parallel injections and / or infusion, wherein one or all compounds are delivered as an injection and / or infusions or a distinct compound is injected and another distinct compound is infused.

[0302] Subject matter of the present invention is a pharmaceutical combination or pharmaceutical composition, wherein Peptide-Gly is applied at a dosage of 0.1-500 pg / kg, preferably 1-400 pg / kg, more preferably 2-300 pg / mU, more preferably 2-300 pg / kg, more preferably 3-200 pg / kg, more preferably 5-100 pg / kg and wherein PAM is applied at a dosage of 1.5 - 54 Units / kg, preferably 1.7-33 Units / kg, more preferably 1.8-26 Units / kg, more preferably 1.9-20.6 Units / kg, more preferably 2-14 Units / kg. In one embodiment 0.1-500 pg / kg of Peptide-Gly are combined with 1.5-54 Units / kg of PAM, more preferably 1-400 pg / kg of Peptide-Gly are combined with 1.7-33 Units / kg PAM, more preferably 2-300 pg / kg Peptide-Gly are combined with 1.8-26 Units / kg PAM, more preferably 3-200 pg / kg of Peptide-Gly are combined with 1.9-20.6 Units / kg of PAM. In the preferred embodiment 5-100 pg / kg of Peptide-Gly are combined with 2-14 Units / kg of PAM.

[0303] Subject matter of the present invention is a pharmaceutical combination or pharmaceutical composition, wherein said combination or composition comprises in addition Vitamin C and wherein Vitamin C applied at a dosage of 1-10000 mg / kg, preferably 2-8000 mg / kg, more preferably 3-6000 mg / kg, more preferably 4-4000 mg / kg, more preferably 5-2000 mg / kg, more preferably 10-1000 mg / kg.

[0304] In one embodiment 0.1-500 pg / kg of Peptide-Gly and 1.5-54 Units / kg of PAM are combined with 1- 10000 mg / kg, preferably 2-8000 mg / kg, more preferably 3-6000 mg / kg, more preferably 4-4000 mg / kg, more preferably 5-2000 mg / kg, more preferably 10-1000 mg / kg of Vitamin C.

[0305] In one embodiment 1-400 pg / kg of Peptide-Gly and 1.7-33 Units / kg PAM are combined with 1-10000 mg / kg, preferably 2-8000 mg / kg, more preferably 3-6000 mg / kg, more preferably 4-4000 mg / kg, more preferably 5-2000 mg / kg, more preferably 10-1000 mg / kg of Vitamin C.

[0306] In one embodiment 2-300 pg / kg of Peptide-Gly and 1.8-26 Units / kg PAM are combined with 1-10000 mg / kg, preferably 2-8000 mg / kg, more preferably 3-6000 mg / kg, more preferably 4-4000 mg / kg, more preferably 5-2000 mg / kg, more preferably 10-1000 mg / kg of Vitamin C.

[0307] In one embodiment 3-200 pg / kg Peptide-Gly and 1.9-20.6 Units / kg PAM are combined with 1-10000 mg / kg, preferably 2-8000 mg / kg, more preferably 3-6000 mg / kg, more preferably 4-4000 mg / kg, more preferably 5-2000 mg / kg, more preferably 10-1000 mg / kg of Vitamin C.

[0308] In one embodiment 5-100 pg / kg of Peptide-Gly with 2-14 Units / kg of PAM are combined with 1-10000 mg / kg, preferably 2-8000 mg / kg, more preferably 3-6000 mg / kg, more preferably 4-4000 mg / kg, more preferably 5-2000 mg / kg, more preferably 10-1000 mg / kg of Vitamin C.

[0309] In the preferred embodiment 5-100 pg / kg of Peptide-Gly are combined with 2-14 Units / kg of PAM and 10-1000 mg / kg of Vitamin C.

[0310] Subject matter of the present invention is Pharmaceutical combination or pharmaceutical composition, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) wherein said PAM is modified via amino acid manipulations, preferably via fusion to Albumins, e.g. serum Albumin or recombinant serum Albumin, more preferably via non covalent binding to serum Albumin due to a conjugated fatty acid chain to PAM, mor preferably via fusion with IgG Fc regions or Transferrin, preferably modified to act as PEG-based carrier prodrug, most preferably via post- translational modifications attaching natural or synthetic polymers, whereas the natural or synthetic polymer to be used is HAP, preferably ELP, more preferably PAS, more preferably PSA, more preferably GLK, more preferably XTEN and most preferably PEG.

[0311] An embodiment of the present invention is a pharmaceutical combination or pharmaceutical composition, wherein ADM-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) wherein said PAM is modified via amino acid manipulations, preferably via fusion to Albumins, e.g. serum Albumin or recombinant serum Albumin, more preferably via non covalent binding to serum Albumin due to a conjugated fatty acid chain to PAM, mor preferably via fusion with IgG Fc regions or Transferrin, preferably modified to act as PEG-based carrier prodrug, most preferably via post-translational modifications attaching natural or synthetic polymers, whereas the natural or synthetic polymer to be used is HAP, preferably ELP, more preferably PAS, more preferably PSA, more preferably GLK, more preferably XTEN and most preferably PEG.

[0312] A further embodiment of the present invention is a pharmaceutical combination or pharmaceutical composition, wherein GLP-l-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) wherein said PAM is modified via amino acid manipulations, preferably via fusion to Albumins, e.g. serum Albumin or recombinant serum Albumin, more preferably via non covalent binding to serum Albumin due to a conjugated fatty acid chain to PAM, mor preferably via fusion with IgG Fc regions or Transferrin, preferably modified to act as PEG-based carrier prodrug, most preferably via post-translational modifications attaching natural or synthetic polymers, whereas the natural or synthetic polymer to be used is HAP, preferably ELP, more preferably PAS, more preferably PSA, more preferably GLK, more preferably XTEN and most preferably PEG.

[0313] Subject matter of the present invention is a method for a long-lasting elevation of amidated peptide hormones like Peptide-Gly in the circulation by the combinatory use of its glycine-extended precursors with the activator PAM and in one embodiment with the Cofactor Vitamin C, as well as the use of the method to treat and / or prevent an acute and / or chronic disease with such combinations. The PAM enzyme is present in circulation and is capable of amidation in circulation.

[0314] The present invention encompasses a compound or a combination of compounds that is suitable for use in the pharmaceutical industry. This compound includes not only the compound itself, but also a pharmaceutically acceptable salt of the compound and a solvent that is considered safe for pharmaceutical use. It is worth noting that the pharmaceutically acceptable salt and solvate of the compound are not limited to any specific type, but the salt or solvate exemplified above is preferred. The compound, when in the form of a salt or solvate, can be used in various pharmaceutical applications. The compound shall mean a combination of PAM and a Peptide-Gly and / or a combination of Peptide- Gly with PAM and Vitamin C.

[0315] One embodiment of the present invention is a compound with a composition in which the inactive Peptide-Gly, which is a natural precursor of an active Peptide-NH2, and recombinant PAM (peptidyl- glycine alpha amidating monooxygenase), which is the natural activator of Peptide-Gly to form Peptide- NH2 are administered in combination. Therefore, the compound of one aspect of the present invention is safe and has low toxicity. Accordingly, pharmaceuticals containing this embodiment of the present invention's compound or its pharmaceutically acceptable salts or solvates as active ingredients can be applied to various subjects requiring prevention or treatment of the symptoms, diseases, and / or disorders.

[0316] Preferred subjects include human or non-human mammals (e.g. pigs, dogs, cows, rats, mice, guinea pigs, rabbits, chickens, sheep, cats, monkeys, orangutans, or chimpanzees), either as experimental subjects or patients.

[0317] By administering the pharmaceutical containing this embodiment of the present invention's compound to the subject, the symptoms, diseases, and / or disorders in the subject can be prevented or treated.

[0318] Throughout the specification the term “ADM-Gly” includes all splice variants of ADM-Gly if not stated otherwise.

[0319] As used herein, the terms "comprising" and "including" or grammatical variants thereof are to be taken as specifying at least the stated features, integers, steps or components but do not preclude the addition of one or more additional features, integers, steps, components or groups thereof. This term encompasses the terms "consisting of and "consisting essentially of that are understood to specify only the stated feature, integers, steps or components to the exclusion of any additional features.

[0320] Thus, the terms "comprising / including / having" mean that any further component (or likewise features, integers, steps and the like) can / may be present.

[0321] The term "consisting” of means that no further component (or likewise features, integers, steps and the like) is present.

[0322] The term "method" refers to manners, means, techniques and procedures for accomplishing a given task including, but not limited to, those manners, means, techniques and procedures either known to, or readily developed from known manners, means, techniques and procedures by practitioners of the chemical, biological and biophysical arts.

[0323] The present invention is further described by reference to the following non-limiting figures. Figure Description

[0324] Fig. 1A: Representative calibration curve of the ADM-Gly Assay with synthetic human 1-53 ADM-Gly

[0325] Fig. IB: Frequency distribution (histogram) of ADM-Gly (pg / mL) in self-reported healthy individuals (n=157)

[0326] Fig. 2A: Representative calibration curve of PAM Amidating Activity Assay (PAM-AMA).

[0327] Fig. 2B: Frequency distribution (histogram) of PAM-AMA in self-reported healthy individuals (n=120) Fig. 2C: Correlation of PAM-AMA in matrix duplets (Li -heparin and serum) from self-reported healthy individuals (n=20)

[0328] Fig. 3A: Typical calibration curves of PAM sandwich immunoassay with recombinant PAM as calibration material in set-up utilizing antibodies against conformational peptides. Solid phase: antibody directed against PAL subunit, tracer: antibody directed against PHM subunit.

[0329] Fig. 3B: Frequency distribution (histogram) of PAM concentration (conformational PAM-LIA) in healthy individuals (n=4106).

[0330] Fig. 4: Bio-ADM levels in rats before (=BL) and after the infusion of ADM-Gly (black bars), ADM- Gly in combination with Vitamin C (open bars), ADM-Gly with PAM (grey bars) and ADM-Gly in combination with Vitamin C and PAM (hatched bars).

[0331] Fig. 5: Determination of human 1-53 ADM-Gly half-life in circulation of rats. The half-life of ADM- Gly of 25.06 minutes was determined with the exponential one-phase decay algorithm of GraphPad Prism software Version 7.0.

[0332] Fig. 6: A representative SDS-PAGE of recombinant human PAM after and before PEGylation.

[0333] Fig. 7: Effect of an intraperitoneal treatment in a murine CLP model with a combination of ADM-Gly and PEG-PAM 24h in advance to model induction on levels of bio-ADM before surgical model induction (left, timepoints BL= baseline and 24h post treatment) and after surgical model induction (t=48h post treatment). Effect of treatment on Blood-Brain Barrier integrity measured as Evans-Blue Extravasation in brain tissue homogenates. Right side: Exemplary brains are shown for the SHAM and CLP arms of the study.

[0334] Fig. 8: The product of the Adrenomedullin gene, the Proadrenomedullin encodes for four peptide hormones and undergoes several enzymatic cleavage steps mainly by prohormone convertases (PCs) to release said peptide hormones. PAMP-Gly and Adrenomedullin-Gly are inactive and can be activated via Amidation into their active forms, PAMP-NH2 and Adrcnomcdullin-NtL (bio-ADM) respectively.

[0335] Fig. 9: Schematic representation of PAM Isoform 1. Black bold arrows indicate cleavage-sites at double- basic amino-acids. Fig. 10: Left: comparative activity analysis of pegylated or xtenylated (i.e. modified with XTEN) and unmodified PAM enzymes. Right: comparative analysis of pegylated or xtenylated and unmodified PAM enzymes on an SDS-PAGE

[0336] The following embodiments form also part of the present invention:

[0337] 1. Peptide-Gly for use in treatment of a disease in a subject, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein PAM and / or Peptide-Gly may be in modified or unmodified form.

[0338] 2. Peptide-Gly for use in treatment of a disease in a subject according to embodiment 1, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein Peptide-Gly is selected from the group comprising

[0339] • from the group of Adrenomedullin family of peptides: Adrenomedullin-2-Gly; Intermedin- Short-Gly and Proadrenomedullin N-20 terminal peptide-Gly,

[0340] • from the Bombesin / neurimedin-B / Ranatensin family of peptides: Gastrin-releasing peptide- Gly; Neuromedin-C-Gly; Neuromedin-B-Gly and Nouromedin-B-32-Gly,

[0341] • from the Calcitonin family of peptides: Calcitonin-Gly; Calcitonin gene-related peptide (CGRP) 1-Gly; CGRP 2-Gly and Islet amyloid polypeptide-Gly,

[0342] • from the Chromogranin / Secretogranin family of peptides: Chromogranin-A-Gly; ER-37-Gly; GR-44-Gly and Pancreastatin-Gly,

[0343] • from the FMRF-amide related peptide family of peptides: Neuropeptide AF-Gly; Neuropeptide FF-Gly; Neuropeptide SF-Gly Neuropeptide NPSF-Gly; Neuropeptide RFRP-l-Gly; Prolactinreleasing peptide PrRP20-Gly and Prolactin-releasing peptide PrRP31-Gly,

[0344] • from the Gastrin / Cholecystokinin family of peptides: Cholecystokinin -Gly; Cholecystokinin - 12-Gly; Cholecystokinin -18-Gly; Cholecystokinin -25-Gly; Cholecystokinin -33-Gly; Cholecystokinin -39-Gly; Cholecystokinin -5-Gly; Cholecystokinin -58-Gly; Cholecystokinin - 8-Gly; Cholecystokinin -7-Gly; Big-gastrin-Gly; Gastrin-Gly; Gastrin- 14-Gly; Gastrin-52-Gly; Gastrin-6-Gly and Gastrin-71 -Gly,

[0345] • from the Glucagon family of peptides: Glucagon-like peptide (GLP) 1-Gly; GLP-l-Gly (7-37); Pituitary adenylate cyclase-activating polypeptide (PACP) 27-Gly; PACAP 38-Gly; Secretin- Gly; Somatoliberin-Gly; Intestinal Peptide PHM-27-Gly; Intestinal Peptide PHV-42-Gly and Vasoactive intestinal Peptide (VIP)-Gly,

[0346] • from the gonadotropin-releasing hormone family of peptides: Ginadoliberin- 1-Gly; Gonadoliberin-2-Gly; Progonadoliberin- 1-Gly and Progonadoliberin-2-Gly, • from the Kisspeptin family of peptides: Kisspeptin-10-Gly; Kisspeptin-13-Gly; Kisspeptin-14- Gly; Metastin-Gly and Metastasis-suppressor KISS-l-Gly,

[0347] • from the Melanin-concentrating hormone family of peptides: Neuropeptide-glutamic acid- isoleucine-Gly and Pro-MCH-Gly,

[0348] • from the Neuromedin family of peptides: Neuromedin-S-Gly and Neuromedin-U-25-Gly,

[0349] • from the Neuropeptide-Y (NPY) family of peptides: Neuropeptide Y-Gly; Pancreatic hormone- Gly; Peptide YY-Gly and Peptide YY( 3-36)-Gly,

[0350] • from the opioid family of peptides: Deltrophin I-Gly,

[0351] • from the Orexin family of peptides: Orexin-A-Gly and Orexin-B-Gly,

[0352] • from the Proopiomelanocortin (POMC) family of peptides: Corticotropin-Gly; alpha- Melanotropin-Gly; gamma-Melanotropin-Gly and N-Terminal peptide of proopiomelanocortin- Gly (NPP-Gly),

[0353] • from the RFamide neuropeptide family of peptides: Orexigenic neuropeptide QRF-Gly (QRF- Gly),

[0354] • from the Sauvagine / corticotropin-releasing factor / urotensin I family of peptides: Urocortin-Gly and Urocortin-3-Gly,

[0355] • from the Tachykinin family of peptides: Substance P-Gly (RPKPQQFFGLMG) and Substance P-Gly (RPKPQFFGLMG); Neurokinin A-Gly; Neuropeptide K-Gly and Neurokinin-B-Gly,

[0356] • from the Thyrotropin Releasing Hormone (TRH) family of peptides: pro-TRH-Gly and TRH- Gly,

[0357] • from the Vasopressin / Oxytocin family of peptides: Oxytocin-Gly and Arg-Vasopressin-Gly,

[0358] • from the VGF nerve growth factor inducible protein (VGF) family of peptides: Antimicrobial peptide VGF-Gly (aa554-578) and Neurosecretory protein VGF-Gly.

[0359] 3. Peptide-Gly for use in treatment of a disease in a subject according to embodiments 1 or 2, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein Peptide-Gly is Adrenomedullin (ADM)-Gly and wherein ADM-Gly may be in modified or unmodified form.

[0360] 4. Peptide-Gly for use in treatment of a disease in a subject according to embodiments 1 to 3, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), wherein said subject is in need of elevated level of Peptide-bTT (Peptide-amide) in the circulation wherein elevated level means by at least 10 pg / mL, preferably by at least 20 pg / mL, more preferably by at least 30 pg / mL, more preferably by at least 40 pg / mL, most preferably by at least 50 pg / mL. Peptide-Gly for use in treatment of a disease in a subject according to embodiments 1 to 4, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), wherein said disease is characterized by a level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or a Peptide-Gly / corresponding Peptide-amide ratio above a threshold in a sample of bodily fluid of said subject and / or a Peptide- amide level below a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or a Peptide- amide level below a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof above a threshold and / or a Peptide- amide level above a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or a Peptide- amide level above a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof above a threshold. Peptide-Gly for use in treatment of a disease in a subject according to any of embodiments 1 to 5, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), and wherein said disease is selected from the group comprising pulmonary hypertension and hypertension, type-2 diabetes, inflammatory bowel disease and / or Crohn's disease, sepsis and / or septic shock, cerebral infarction, mild cognitive impairment (MCI) and Alzheimer’s Disease (AD). Peptide-Gly for use in treatment of a disease in a subject according to any of the preceding embodiments, wherein Peptide-Gly is GLP-l-Gly (SEQ ID No.: 39-40), wherein GLP-l-Gly (SEQ ID No.: 39-40) is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), and wherein said disease is type-2 diabetes. Peptide-Gly for use in treatment of a disease in a subject according to any of embodiments 1 to 7, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and in addition is to be used with Vitamin C. Peptide-Gly for use in treatment of a disease in a subject according to any of embodiments 1 to

[0361] 8, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein Peptide-Gly and PAM are to be administered consecutively, or Peptide-Gly and PAM are to be administered simultaneously. Peptide-Gly for use in treatment of a disease in a subject according to any of embodiments 1 to 9, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein the ratio of Peptide-Gly and the corresponding Peptideamide in a bodily fluid is elevated over 1.5, preferably elevated over 1.6, more preferably elevated over 1.7, more preferably is elevated over 1.8, more preferable is elevated over 1.9, more preferably is higher than 2 and / or the concentration of PAM (SEQ ID No. 1-10) in a bodily fluid is equal or below 100 ng / mL (100 ng / mL to 0 ng / mL), preferably equal or below 90 ng / mL (90 ng / mL to 0 ng / mL), more preferably equal or below 80 ng / mL (80 ng / mL to 0 ng / mL), more preferably equal or below 60 ng / mL (60 ng / mL to 0 ng / mL), more preferably equal or below 50 ng / mL (50 ng / mL to 0 ng / mL) and most preferably equal or below 40 ng / mL (40 ng / mL to 0 ng / mL)and / or the activity of PAM (SEQ ID No. 1-10) in a bodily fluid is equal or below 24 Units / L, more preferably equal or below 20 Units / L, more preferably equal or below 16 Units / L, more preferably equal or below 14 Units / L, and most preferably equal or below 10 Units / L and / or

[0362] Peptide-amide level is equal or below 20 pg / mL, preferably equal or below 15 pg / mL, preferably equal or below 10 pg / mL, preferably equal or below 5 pg / mL and / or

[0363] Peptide- is equal or above 15 pg / mL, preferably equal or above 20 pg / mL, preferably equal or above 30 pg / mL, preferably is equal or above 50 pg / mL, and / or

[0364] Peptide-amide level is equal or above 70 pg / mL, irrespective of the status of PAM and / or Peptide-Gly, and / or the concentration of PAM (SEQ ID No.: 1-10) is equal or above 80 ng / mL, preferably equal or above 100 ng / mL, irrespective of the corresponding Peptide-amide and Peptide-Gly status, and / or the activity of PAM (SEQ ID No.: 1-10) is equal or above 14 Units / L, preferably equal or above 20 units / L, preferably equal or above 24 units / L, irrespective of a corresponding Peptide-amide and Peptide-Gly level. Peptide-Gly for use in treatment of a disease in a subject according to any of embodiments 1 to

[0365] 10, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein Peptide-Gly is applied at a dosage of 0.1-500 pg / kg, preferably 1-400 pg / kg, more preferably 2-300 pg / mL, more preferably 2-300 pg / kg, more preferably 3-200 pg / kg, more preferably 5-100 pg / kg and wherein PAM is applied at a dosage of 1.5 - 54 Units / kg, preferably 1.7-33 Units / kg, more preferably 1.8-26 Units / kg, more preferably 1.9-20.6 Units / kg, more preferably 2-14 Units / kg Peptide-Gly for use in treatment of a disease in a subject according to any of embodiments 1 to

[0366] 11, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and in addition is to be used with Vitamin C and wherein Vitamin C is applied at a dosage of 1-10000 mg / kg, preferably 2-8000 mg / kg, more preferably 3-6000 mg / kg, more preferably 4-4000 mg / kg, more preferably 5-2000 mg / kg, more preferably 10-1000 mg / kg. Peptide-Gly for use in treatment of a disease in a subject according to any of embodiments 1 to

[0367] 12, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) wherein said PAM is modified via amino acid manipulations, preferably via fusion to Albumins, e.g. serum Albumin or recombinant serum Albumin, more preferably via non covalent binding to serum Albumin due to a conjugated fatty acid chain to PAM, mor preferably via fusion with IgG Fc regions or Transferrin, more preferably modified to act as PEG-based carrier prodrug, most preferably via post-translational modifications attaching natural or synthetic polymers, whereas the natural or synthetic polymer to be used is HAP, preferably EEP, more preferably PAS, more preferably PSA, more preferably GLK, more preferably XTEN and most preferably PEG. Peptide-Gly for use in treatment of a disease in a subject according to any of embodiments 1 to

[0368] 13, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) wherein said PAM is modified by pegylation. Pharmaceutical combination or pharmaceutical composition comprising Peptide-Gly and peptidylglycine a-amidating monooxygenase (PAM), wherein said combination may be a kits of part combination with at least two pharmaceutical compositions, wherein one of which comprises Peptide-Gly and wherein the other comprises PAM or wherein said pharmaceutical composition comprises both Peptide-Gly and peptidylglycine a-amidating monooxygenase (PAM), and wherein PAM and / or Peptide-Gly may be in modified or unmodified form. Pharmaceutical combination or pharmaceutical composition according to embodiment 15, wherein said combination or composition comprises in addition Vitamin C. 17. Pharmaceutical combination or pharmaceutical composition according to any of embodiment 15 or 16, wherein Peptide-Gly is applied at a dosage of 0.1-500 pg / kg, preferably 1-400 pg / kg, more preferably 2-300 pg / mL, more preferably 2-300 pg / kg, more preferably 3-200 pg / kg, more preferably 5-100 pg / kg and wherein PAM is applied at a dosage of 1.5 - 54 Units / kg, preferably 1.7-33 Units / kg, more preferably 1.8-26 Units / kg, more preferably 1.9-20.6Units / kg, more preferably 2-14.0 Units / kg

[0369] 18. Pharmaceutical combination or pharmaceutical composition according to any of embodiments 15 or 17, wherein said combination or composition comprises in addition Vitamin C and wherein Vitamin C is applied at a dosage of 1-10000 mg / kg, preferably 2-8000 mg / kg, more preferably 3-6000 mg / kg, more preferably 4-4000 mg / kg, more preferably 5-2000 mg / kg, more preferably 10-1000 mg / kg.

[0370] 19. Pharmaceutical combination or pharmaceutical composition according to any of embodiment 15 or 18, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) wherein said PAM is modified via amino acid manipulations, preferably via fusion to Albumins, e.g. serum Albumin or recombinant serum Albumin, more preferably via non covalent binding to serum Albumin due to a conjugated fatty acid chain to PAM, mor preferably via fusion with IgG Fc regions or Transferrin, preferably modified to act as PEG-based carrier prodrug, most preferably via post-translational modifications attaching natural or synthetic polymers, whereas the natural or synthetic polymer to be used is HAP, preferably EEP, more preferably PAS, more preferably PSA, more preferably GLK, more preferably XTEN and most preferably PEG.

[0371] 20. A method of treating a disease in a subject, said method comprising administering to said subject an effective amount of Peptide-Gly, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein PAM and / or Peptide-Gly may be in modified or unmodified form.

[0372] 21. A method of treating a disease in a subject, said method comprising administering to said subject an effective amount of Peptide-Gly according to embodiment 20, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein Peptide-Gly is selected from the group comprising

[0373] • from the group of Adrenomedullin family of peptides: Adrenomedullin-2-Gly; Intermedin- Short-Gly and Proadrenomedullin N-20 terminal peptide-Gly,

[0374] • from the Bombesin / neurimedin-B / Ranatensin family of peptides: Gastrin-releasing peptide- Gly; Neuromedin-C-Gly; Neuromedin-B-Gly and Nouromedin-B-32-Gly, • from the Calcitonin family of peptides: Calcitonin-Gly; Calcitonin gene-related peptide (CGRP) 1-Gly; CGRP 2-Gly and Islet amyloid polypeptide-Gly,

[0375] • from the Chromogranin / Secretogranin family of peptides: Chromogranin-A-Gly; ER-37-Gly; GR-44-Gly and Pancreastatin-Gly,

[0376] • from the FMRF-amide related peptide family of peptides: Neuropeptide AF-Gly; Neuropeptide FF-Gly; Neuropeptide SF-Gly Neuropeptide NPSF-Gly; Neuropeptide RFRP-l-Gly; Prolactinreleasing peptide PrRP20-Gly and Prolactin-releasing peptide PrRP31-Gly,

[0377] • from the Gastrin / Cholecystokinin family of peptides: Cholecystokinin-Gly; Cholecystokinin-

[0378] 12-Gly; Cholecystokinin- 18-Gly; Cholecystokinin-25 -Gly; Cholecystokinin-33 -Gly;

[0379] Cholecystokinin-39-Gly; Cholecystokinin-5 -Gly; Cholecystokinin-58-Gly; Cholecystokinin-8- Gly; Cholecystokinin-7-Gly; Big-gastrin-Gly; Gastrin-Gly; Gastrin- 14-Gly; Gastrin-52-Gly; Gastrin-6-Gly and Gastrin-71 -Gly,

[0380] • from the Glucagon family of peptides: Glucagon-like peptide (GLP) 1-Gly; GLP-l-Gly (7-37); Pituitary adenylate cyclase-activating polypeptide (PACP) 27-Gly; PACAP 38-Gly; Secretin- Gly; Somatoliberin-Gly; Intestinal Peptide PHM-27-Gly; Intestinal Peptide PHV-42-Gly and Vasoactive intestinal Peptide (VIP)-Gly,

[0381] • from the gonadotropin-releasing hormone family of peptides: Ginadoliberin- 1-Gly; Gonadoliberin-2-Gly; Progonadoliberin- 1-Gly and Progonadoliberin-2-Gly,

[0382] • from the Kisspeptin family of peptides: Kisspeptin-10-Gly; Kisspeptin-13-Gly; Kisspeptin- 14- Gly; Metastin-Gly and Metastasis-suppressor KISS- 1-Gly,

[0383] • from the Melanin-concentrating hormone family of peptides: Neuropeptide-glutamic acid- isoleucine-Gly and Pro-MCH-Gly,

[0384] • from the Neuromedin family of peptides: Neuromedin-S-Gly and Neuromedin-U-25-Gly,

[0385] • from the Neuropeptide-Y (NPY) family of peptides: Neuropeptide Y-Gly; Pancreatic hormone- Gly; Peptide YY-Gly and Peptide YY( 3-36)-Gly,

[0386] • from the opioid family of peptides: Deltrophin I-Gly,

[0387] • from the Orexin family of peptides: Orexin-A-Gly and Orexin-B-Gly,

[0388] • from the Proopiomelanocortin (POMC) family of peptides: Corticotropin-Gly; alpha- Melanotropin-Gly; gamma-Melanotropin-Gly and N-Terminal peptide of proopiomelanocortin- Gly (NPP-Gly),

[0389] • from the RFamide neuropeptide family of peptides: Orexigenic neuropeptide QRF-Gly (QRF- Gly),

[0390] • from the Sauvagine / corticotropin-releasing factor / urotensin I family of peptides: Urocortin-Gly and Urocortin-3-Gly,

[0391] • from the Tachykinin family of peptides: Substance P-Gly (RPKPQQFFGLMG) and Substance P-Gly (RPKPQFFGLMG); Neurokinin A-Gly; Neuropeptide K-Gly and Neurokinin-B-Gly, • from the Thyrotropin Releasing Hormone (TRH) family of peptides: pro-TRH-Gly and TRH- Gly,

[0392] • from the Vasopressin / Oxytocin family of peptides: Oxytocin-Gly and Arg-Vasopressin-Gly,

[0393] • from the VGF nerve growth factor inducible protein (VGF) family of peptides: Antimicrobial peptide VGF-Gly (aa554-578) and Neurosecretory protein VGF-Gly.

[0394] 22. A method of treating a disease in a subject, said method comprising administering to said subject an effective amount of Peptide-Gly according to embodiments 20 or 21, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein Peptide Gly is ADM-Gly and wherein ADM-Gly may be in modified or unmodified form.

[0395] 23. A method of treating a disease in a subject, said method comprising administering to said subject an effective amount of Peptide-Gly according to embodiment 20 to 22, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), wherein said subject is in need of elevated level of Peptide-fMT (Peptide-amide) in the circulation wherein elevated level means by at least 10 pg / mL, preferably by at least 20 pg / mL, more preferably by at least 30 pg / mL, more preferably by at least 40 pg / mL, most preferably by at least 50 pg / mL.

[0396] 24. A method of treating a disease in a subject, said method comprising administering to said subject an effective amount of Peptide-Gly according to embodiment 20 to 23, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), wherein said disease is characterized by a level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or a Peptide-Gly / corresponding Peptide-amide ratio above a threshold in a sample of bodily fluid of said subject and / or a Peptide-amide level below a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or a Peptide-amide level below a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof above a threshold and / or a Peptide-amide level above a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or a Peptide-amide level above a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof above a threshold. A method of treating a disease in a subject, said method comprising administering to said subject an effective amount of Peptide-Gly according to any of embodiments 20 to 24, wherein Peptide- Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), and wherein said disease is selected from the group comprising pulmonary hypertension and hypertension, type-2 diabetes, inflammatory bowel disease and / or Crohn's disease, sepsis and / or septic shock, cerebral infarction, mild cognitive impairment (MCI) and Alzheimer’s Disease (AD). A method of treating a disease in a subject, said method comprising administering to said subject an effective amount of Peptide-Gly according to any of embodiments 20 to 25, wherein Peptide- Gly is GLP-l-Gly (SEQ ID No.: 39-40), wherein GLP-l-Gly (SEQ ID No.: 39-40) is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), and wherein said disease is type-2 diabetes. A method of treating a disease in a subject, said method comprising administering to said subject an effective amount of Peptide-Gly according to any of embodiments 20 to 26, wherein Peptide- Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and in addition is to be used with Vitamin C. A method of treating a disease in a subject, said method comprising administering to said subject an effective amount of Peptide-Gly according to any of embodiments 20 to 27, wherein Peptide- Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein Peptide-Gly and PAM are to be administered consecutively, or Peptide-Gly and PAM are to be administered simultaneously. A method of treating a disease in a subject, said method comprising administering to said subject an effective amount of Peptide-Gly according to any of embodiments 20 to 28, wherein the ratio of Peptide-Gly and the corresponding Peptide-amide in a bodily fluid is elevated over 1.5, preferably elevated over 1.6, more preferably elevated over 1.7, more preferably is elevated over 1.8, more preferable is elevated over 1.9, more preferably is higher than 2 and / or the concentration of PAM (SEQ ID No. 1-10) in a bodily fluid is equal or below 100 ng / mL (100 ng / mL to 0 ng / mL), preferably equal or below 90 ng / mL (90 ng / mL to 0 ng / mL), more preferably equal or below 80 ng / mL (80 ng / mL to 0 ng / mL), more preferably equal or below 60 ng / mL (60 ng / mL to 0 ng / mL), more preferably equal or below 50 ng / mL (50 ng / mL to 0 ng / mL) and most preferably equal or below 40 ng / mL (40 ng / mL to 0 ng / mL) and / or the activity of PAM (SEQ ID No. 1-10) in a bodily fluid is equal or below 24 Units / L, more preferably equal or below 20 Units / L, more preferably equal or below 16 Units / L, more preferably equal or below 14 Units / L, and most preferably equal or below 10 Units / L and / or

[0397] Peptide-amide level is equal or below 20 pg / mL, preferably equal or below 15 pg / mL, preferably equal or below 10 pg / mL, preferably equal or below 5 pg / mL and / or

[0398] Peptide-Gly is equal or above 15 pg / mL, preferably equal or above 20 pg / mL, preferably equal or above 30 pg / mL, preferably is equal or above 50 pg / mL, and / or

[0399] Peptide-amide level is equal or above 70 pg / mL, irrespective of the status of PAM and / or Peptide-Gly, and / or the concentration of PAM (SEQ ID No.: 1-10) is equal or above 80 ng / mL, preferably equal or above 100 ng / mL, irrespective of the corresponding Peptide-amide and Peptide-Gly status, and / or the activity of PAM (SEQ ID No.: 1-10) is equal or above 14 Units / L, preferably equal or above 20 units / L, preferably equal or above 24 units / L, irrespective of a corresponding Peptide-amide and Peptide-Gly level. A method of treating a disease in a subject, said method comprising administering to said subject an effective amount of Peptide-Gly according to any of embodiments 20 to 29, wherein Peptide- Gly is applied at a dosage of 0.1-500 pg / kg, preferably 1-400 pg / kg, more preferably 2-300 pg / mL, more preferably 2-300 pg / kg, more preferably 3-200 pg / kg, more preferably 5-100 pg / kg and wherein PAM is applied at a dosage of 1.5 - 54 Units / kg, preferably 1.7-33 Units / kg, more preferably 1.8-26 Units / kg, more preferably 1.9-20.6Units / kg, more preferably 2-14.0 Units / kg. A method of treating a disease in a subject, said method comprising administering to said subject an effective amount of Peptide-Gly according to any of embodiments 20 to 30, wherein Peptide- Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and in addition is to be used with Vitamin C and wherein Vitamin C is applied at a dosage of 1- 10000 mg / kg, preferably 2-8000 mg / kg, more preferably 3-6000 mg / kg, more preferably 4-4000 mg / kg, more preferably 5-2000 mg / kg, more preferably 10-1000 mg / kg.

[0400] 32. A method of treating a disease in a subject, said method comprising administering to said subject an effective amount of Peptide-Gly according to any of embodiments 20 to 31, wherein Peptide- Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) wherein said PAM is modified via amino acid manipulations, preferably via fusion to Albumins, e.g. serum Albumin or recombinant serum Albumin, more preferably via non covalent binding to serum Albumin due to a conjugated fatty acid chain to PAM, mor preferably via fusion with IgG Fc regions or Transferrin, preferably modified to act as PEG-based carrier prodrug, most preferably via post-translational modifications attaching natural or synthetic polymers, whereas the natural or synthetic polymer to be used is HAP, preferably ELP, more preferably PAS, more preferably PSA, more preferably GLK, more preferably XTEN and most preferably PEG.

[0401] 33. A method of treating a disease in a subject, said method comprising administering to said subject an effective amount of Peptide-Gly according to any of embodiments 20 to 32, wherein Peptide- Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) wherein said PAM is modified by pegylation.

[0402] Moreover, the following embodiments also form part of the present invention:

[0403] 1. ADM-Gly for use in treatment of a disease in a subject, wherein ADM-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein PAM and / or ADM-Gly may be in modified or unmodified form.

[0404] 2. ADM-Gly for use in treatment of a disease in a subject according to embodiment 1, wherein ADM-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), wherein said subject is in need of an elevated level of ADM-NH2 (ADM-amide) in the circulation wherein elevated level means by at least 10 pg / mL, preferably by at least 20 pg / mL, more preferably by at least 30 pg / mL, more preferably by at least 40 pg / mL, most preferably by at least 50 pg / mL.

[0405] 3. ADM-Gly for use in treatment of a disease in a subject according to embodiments 1 or 2, wherein ADM-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), wherein said disease is characterized by a level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or an ADM-Gly / ADM-amide ratio above a threshold in a sample of bodily fluid of said subject and / or an ADM- amide level below a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or an ADM- amide level below a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof above a threshold and / or an ADM- amide level above a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or an ADM- amide level above a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof above a threshold. ADM-Gly for use in treatment of a disease in a subject according to any of embodiments 1 to

[0406] 3, wherein ADM-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), and wherein said disease is selected from the group comprising pulmonary hypertension and hypertension, type-2 diabetes, inflammatory bowel disease and / or Crohn's disease, sepsis and / or septic shock, cerebral infarction, mild cognitive impairment (MCI) and Alzheimer’s Disease (AD). ADM-Gly for use in treatment of a disease in a subject according to any of embodiments 1 to

[0407] 4, wherein ADM-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and in addition is to be used with Vitamin C. ADM-Gly for use in treatment of a disease in a subject according to any of embodiments 1 to

[0408] 5, wherein ADM-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein ADM-Gly and PAM are to be administered consecutively, or ADM-Gly and PAM are to be administered simultaneously. ADM-Gly for use in treatment of a disease in a subject according to any of embodiments 1 to

[0409] 6, wherein ADM-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein the ratio of ADM-Gly and the ADM-amide in a bodily fluid is elevated over 1.5, preferably elevated over 1.6, more preferably elevated over 1.7, more preferably is elevated over 1.8, more preferable is elevated over 1.9, more preferably is higher than 2 and / or the concentration of PAM (SEQ ID No. 1-10) in a bodily fluid is equal or below 100 ng / mL (100 ng / mL to 0 ng / mL), preferably equal or below 90 ng / mL (90 ng / mL to 0 ng / mL), more preferably equal or below 80 ng / mL (80 ng / mL to 0 ng / mL), more preferably equal or below 60 ng / mL (60 ng / mL to 0 ng / mL), more preferably equal or below 50 ng / mL (50 ng / mL to 0 ng / mL) and most preferably equal or below 40 ng / mL (40 ng / mL to 0 ng / mL)and / or the activity of PAM (SEQ ID No. 1-10) in a bodily fluid is equal or below 24 Units / L, more preferably equal or below 20 Units / L, more preferably equal or below 16 Units / L, more preferably equal or below 14 Units / L, and most preferably equal or below 10 Units / L and / or

[0410] ADM-amide level is equal or below 20 pg / mL, preferably equal or below 15 pg / mL, preferably equal or below 10 pg / mL, preferably equal or below 5 pg / mL and / or

[0411] ADM- is equal or above 15 pg / mL, preferably equal or above 20 pg / mL, preferably equal or above 30 pg / mL, preferably is equal or above 50 pg / mL, and / or

[0412] ADM-amide level is equal or above 70 pg / mL, irrespective of the status of PAM and / or ADM- Gly, and / or the concentration of PAM (SEQ ID No.: 1-10) is equal or above 80 ng / mL, preferably equal or above 100 ng / mL, irrespective of the ADM-amide and ADM-Gly status, and / or the activity of PAM (SEQ ID No.: 1-10) is equal or above 14 Units / L, preferably equal or above 20 units / L, preferably equal or above 24 units / L, irrespective of a ADM-amide and ADM-Gly level. ADM-Gly for use in treatment of a disease in a subject according to any of embodiments 1 to 7, wherein ADM-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein ADM-Gly is applied at a dosage of 0.1-500 pg / kg, preferably 1-400 pg / kg, more preferably 2-300 pg / mL, more preferably 2-300 pg / kg, more preferably 3-200 pg / kg, more preferably 5-100 pg / kg and wherein PAM is applied at a dosage of 1.5 - 54 Units / kg, preferably 1.7-33 Units / kg, more preferably 1.8-26 Units / kg, more preferably 1.9-20.6 Units / kg, more preferably 2-14 Units / kg. ADM-Gly for use in treatment of a disease in a subject according to any of embodiments 1 to

[0413] 8, wherein ADM-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and in addition is to be used with Vitamin C and wherein Vitamin C is applied at a dosage of 1-10000 mg / kg, preferably 2-8000 mg / kg, more preferably 3-6000 mg / kg, more preferably 4-4000 mg / kg, more preferably 5-2000 mg / kg, more preferably 10-1000 mg / kg. ADM-Gly for use in treatment of a disease in a subject according to any of embodiments 1 to

[0414] 9, wherein ADM-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) wherein said PAM is modified via amino acid manipulations, preferably via fusion to Albumins, e.g. serum Albumin or recombinant serum Albumin, more preferably via non covalent binding to serum Albumin due to a conjugated fatty acid chain to PAM, mor preferably via fusion with IgG Fc regions or Transferrin, more preferably modified to act as PEG-based carrier prodrug, most preferably via post-translational modifications attaching natural or synthetic polymers, whereas the natural or synthetic polymer to be used is HAP, preferably ELP, more preferably PAS, more preferably PSA, more preferably GLK, more preferably XTEN and most preferably PEG. ADM-Gly for use in treatment of a disease in a subject according to any of embodiments 1 to

[0415] 10, wherein ADM-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) wherein said PAM is modified by pegylation. Pharmaceutical combination or pharmaceutical composition comprising ADM-Gly and peptidylglycine a-amidating monooxygenase (PAM), wherein said combination may be a kits of part combination with at least two pharmaceutical compositions, wherein one of which comprises ADM-Gly and wherein the other comprises PAM or wherein said pharmaceutical composition comprises both ADM-Gly and peptidylglycine a-amidating monooxygenase (PAM), and wherein PAM and / or ADM-Gly may be in modified or unmodified form. Pharmaceutical combination or pharmaceutical composition according to embodiment 12, wherein said combination or composition comprises in addition Vitamin C. Pharmaceutical combination or pharmaceutical composition according to any of embodiment 12 or 13, wherein ADM-Gly is applied at a dosage of 0.1-500 pg / kg, preferably 1-400 pg / kg, more preferably 2-300 pg / mL, more preferably 2-300 pg / kg, more preferably 3-200 pg / kg, more preferably 5-100 pg / kg and wherein PAM is applied at a dosage of 1.5 - 54 Units / kg, preferably 1.7-33 Units / kg, more preferably 1.8-26 Units / kg, more preferably 1.9-20.6Units / kg, more preferably 2-14.0 Units / kg 15. Pharmaceutical combination or pharmaceutical composition according to any of embodiments 12 or 14, wherein said combination or composition comprises in addition Vitamin C and wherein Vitamin C is applied at a dosage of 1-10000 mg / kg, preferably 2-8000 mg / kg, more preferably 3-6000 mg / kg, more preferably 4-4000 mg / kg, more preferably 5-2000 mg / kg, more preferably 10-1000 mg / kg.

[0416] 16. Pharmaceutical combination or pharmaceutical composition according to any of embodiment 12 or 15, wherein ADM-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) wherein said PAM is modified via amino acid manipulations, preferably via fusion to Albumins, e.g. serum Albumin or recombinant serum Albumin, more preferably via non covalent binding to serum Albumin due to a conjugated fatty acid chain to PAM, mor preferably via fusion with IgG Fc regions or Transferrin, preferably modified to act as PEG-based carrier prodrug, most preferably via post-translational modifications attaching natural or synthetic polymers, whereas the natural or synthetic polymer to be used is HAP, preferably ELP, more preferably PAS, more preferably PSA, more preferably GLK, more preferably XTEN and most preferably PEG.

[0417] The following embodiments further form part of the present invention:

[0418] 1. A method of treating a disease in a subject, said method comprising administering to said subject an effective amount of ADM-Gly, wherein ADM-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein PAM and / or ADM-Gly may be in modified or unmodified form.

[0419] 2. A method of treating a disease in a subject, said method comprising administering to said subject an effective amount of ADM-Gly according to embodiment 1, wherein ADM-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), wherein said subject is in need of elevated level of ADM-NH2 (ADM-amide) in the circulation wherein elevated level means by at least 10 pg / mL, preferably by at least 20 pg / mL, more preferably by at least 30 pg / mL, more preferably by at least 40 pg / mL, most preferably by at least 50 pg / mL.

[0420] 3. A method of treating a disease in a subject, said method comprising administering to said subject an effective amount of ADM-Gly according to embodiment 1 or 2, wherein ADM-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), wherein said disease is characterized by a level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or an ADM-Gly / corresponding ADM-amide ratio above a threshold in a sample of bodily fluid of said subject and / or an ADM-amide level below a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or an ADM-amide level below a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof above a threshold and / or an ADM-amide level above a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or an ADM-amide level above a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof above a threshold. A method of treating a disease in a subject, said method comprising administering to said subject an effective amount of ADM-Gly according to any of embodiments 1 to 3, wherein ADM-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), and wherein said disease is selected from the group comprising pulmonary hypertension and hypertension, type-2 diabetes, inflammatory bowel disease and / or Crohn's disease, sepsis and / or septic shock, cerebral infarction, mild cognitive impairment (MCI) and Alzheimer’s Disease (AD). A method of treating a disease in a subject, said method comprising administering to said subject an effective amount of ADM-Gly according to any of embodiments 1 to 4, wherein ADM-Gly is GLP-l-Gly (SEQ ID No.: 39-40), wherein GLP-l-Gly (SEQ ID No.: 39-40) is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), and wherein said disease is type-2 diabetes. A method of treating a disease in a subject, said method comprising administering to said subject an effective amount of ADM-Gly according to any of embodiments 1 to 5, wherein ADM-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and in addition is to be used with Vitamin C. A method of treating a disease in a subject, said method comprising administering to said subject an effective amount of ADM-Gly according to any of embodiments 1 to 6, wherein ADM-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein ADM-Gly and PAM are to be administered consecutively, or ADM-Gly and PAM are to be administered simultaneously. A method of treating a disease in a subject, said method comprising administering to said subject an effective amount of ADM-Gly according to any of embodiments 1 to 7, wherein the ratio of ADM-Gly and the corresponding ADM-amide in a bodily fluid is elevated over 1.5, preferably elevated over 1.6, more preferably elevated over 1.7, more preferably is elevated over 1.8, more preferable is elevated over 1.9, more preferably is higher than 2 and / or the concentration of PAM (SEQ ID No. 1-10) in a bodily fluid is equal or below 100 ng / mL (100 ng / mL to 0 ng / mL), preferably equal or below 90 ng / mL (90 ng / mL to 0 ng / mL), more preferably equal or below 80 ng / mL (80 ng / mL to 0 ng / mL), more preferably equal or below 60 ng / mL (60 ng / mL to 0 ng / mL), more preferably equal or below 50 ng / mL (50 ng / mL to 0 ng / mL) and most preferably equal or below 40 ng / mL (40 ng / mL to 0 ng / mL) and / or the activity of PAM (SEQ ID No. 1-10) in a bodily fluid is equal or below 24 Units / L, more preferably equal or below 20 Units / L, more preferably equal or below 16 Units / L, more preferably equal or below 14 Units / L, and most preferably equal or below 10 Units / L and / or

[0421] ADM-amide level is equal or below 20 pg / mL, preferably equal or below 15 pg / mL, preferably equal or below 10 pg / mL, preferably equal or below 5 pg / mL and / or

[0422] ADM-Gly is equal or above 15 pg / mL, preferably equal or above 20 pg / mL, preferably equal or above 30 pg / mL, preferably is equal or above 50 pg / mL, and / or

[0423] ADM-amide level is equal or above 70 pg / mL, irrespective of the status of PAM and / or ADM- Gly, and / or the concentration of PAM (SEQ ID No.: 1-10) is equal or above 80 ng / mL, preferably equal or above 100 ng / mL, irrespective of the corresponding ADM-amide and ADM-Gly status, and / or the activity of PAM (SEQ ID No.: 1-10) is equal or above 14 Units / L, preferably equal or above 20 units / L, preferably equal or above 24 units / L, irrespective of a corresponding ADM-amide and ADM-Gly level. 9. A method of treating a disease in a subject, said method comprising administering to said subject an effective amount of ADM-Gly according to any of embodiments 1 to 8, wherein ADM-Gly is applied at a dosage of 0.1-500 pg / kg, preferably 1-400 pg / kg, more preferably 2-300 pg / mL, more preferably 2-300 pg / kg, more preferably 3-200 pg / kg, more preferably 5-100 pg / kg and wherein PAM is applied at a dosage of 1.5 - 54 Units / kg, preferably 1.7-33 Units / kg, more preferably 1.8-26 Units / kg, more preferably 1.9-20.6Units / kg, more preferably 2-14.0 Units / kg.

[0424] 10. A method of treating a disease in a subject, said method comprising administering to said subject an effective amount of ADM-Gly according to any of embodiments 1 to 9, wherein ADM-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and in addition is to be used with Vitamin C and wherein Vitamin C is applied at a dosage of 1-10000 mg / kg, preferably 2-8000 mg / kg, more preferably 3-6000 mg / kg, more preferably 4-4000 mg / kg, more preferably 5-2000 mg / kg, more preferably 10-1000 mg / kg.

[0425] 11. A method of treating a disease in a subject, said method comprising administering to said subject an effective amount of ADM-Gly according to any of embodiments 1 to 10, wherein ADM-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) wherein said PAM is modified via amino acid manipulations, preferably via fusion to Albumins, e.g. serum Albumin or recombinant serum Albumin, more preferably via non covalent binding to serum Albumin due to a conjugated fatty acid chain to PAM, mor preferably via fusion with IgG Fc regions or Transferrin, preferably modified to act as PEG-based carrier prodrug, most preferably via post-translational modifications attaching natural or synthetic polymers, whereas the natural or synthetic polymer to be used is HAP, preferably EEP, more preferably PAS, more preferably PSA, more preferably GLK, more preferably XTEN and most preferably PEG.

[0426] 12. A method of treating a disease in a subject, said method comprising administering to said subject an effective amount of ADM-Gly according to any of embodiments 1 to 11, wherein ADM-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) wherein said PAM is modified by pegylation.

[0427] The following embodiments form also part of the present invention:

[0428] 1. GLP-l-Gly for use in treatment of a disease in a subject, wherein GLP-l-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein PAM and / or GLP-l-Gly may be in modified or unmodified form.

[0429] 2. GLP-l-Gly for use in treatment of a disease in a subject according to embodiment 1, wherein GLP-l-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), wherein said subject is in need of elevated level of GLP-I-NH2 (GLP-l-amide) in the circulation wherein elevated level means by at least 10 pg / mL, preferably by at least 20 pg / mL, more preferably by at least 30 pg / mL, more preferably by at least 40 pg / mL, most preferably by at least 50 pg / mL. GLP-l-Gly for use in treatment of a disease in a subject according to embodiments 1 or 2, wherein GLP-l-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), wherein said disease is characterized by a level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or an GLP-l-Gly / GLP-l-amide ratio above a threshold in a sample of bodily fluid of said subject and / or a GLP-l-amide level below a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or a GLP-1- amide level below a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof above a threshold and / or a GLP-l-amide level above a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or a GLP-l-amide level above a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof above a threshold. GLP-l-Gly for use in treatment of a disease in a subject according to any of embodiments 1 to 3, wherein GLP-l-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), and wherein said disease is selected from the group comprising pulmonary hypertension and hypertension, type-2 diabetes, inflammatory bowel disease and / or Crohn's disease, sepsis and / or septic shock, cerebral infarction, mild cognitive impairment (MCI) and Alzheimer’s Disease (AD). GLP-l-Gly for use in treatment of a disease in a subject according to any of embodiments 1 to

[0430] 4, wherein GLP-l-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and in addition is to be used with Vitamin C. GLP-l-Gly for use in treatment of a disease in a subject according to any of embodiments 1 to 5, wherein GLP-l-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein GLP-l-Gly and PAM are to be administered consecutively, or GLP-l-Gly and PAM are to be administered simultaneously. Peptide-Gly for use in treatment of a disease in a subject according to any of the preceding embodiments, wherein Peptide-Gly is GLP-l-Gly (SEQ ID No.:39-40), wherein GLP-l-Gly (SEQ ID No.: 39-40) is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), and wherein said disease is type-2 diabetes. GLP-l-Gly for use in treatment of a disease in a subject according to any of embodiments 1 to 7, wherein GLP-l-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein the ratio of GLP-l-Gly and the GLP-1 -amide in a bodily fluid is elevated over 1.5, preferably elevated over 1.6, more preferably elevated over 1.7, more preferably is elevated over 1.8, more preferable is elevated over 1.9, more preferably is higher than 2 and / or the concentration of PAM (SEQ ID No. 1-10) in a bodily fluid is equal or below 100 ng / mL (100 ng / mL to 0 ng / mL), preferably equal or below 90 ng / mL (90 ng / mL to 0 ng / mL), more preferably equal or below 80 ng / mL (80 ng / mL to 0 ng / mL), more preferably equal or below 60 ng / mL (60 ng / mL to 0 ng / mL), more preferably equal or below 50 ng / mL (50 ng / mL to 0 ng / mL) and most preferably equal or below 40 ng / mL (40 ng / mL to 0 ng / mL) and / or the activity of PAM (SEQ ID No. 1-10) in a bodily fluid is equal or below 24 Units / L, more preferably equal or below 20 Units / L, more preferably equal or below 16 Units / L, more preferably equal or below 14 Units / L and most preferably equal or below 10 Units / L and / or

[0431] GLP-1 -amide level is equal or below 20 pg / mL, preferably equal or below 15 pg / mL, preferably equal or below 10 pg / mL, preferably equal or below 5 pg / mL and / or

[0432] GLP-1 is equal or above 15 pg / mL, preferably equal or above 20 pg / mL, preferably equal or above 30 pg / mL, preferably is equal or above 50 pg / mL, and / or

[0433] GLP-1 -amide level is equal or above 70 pg / mL, irrespective of the status of PAM and / or GLP- l-Gly, and / or the concentration of PAM (SEQ ID No.: 1-10) is equal or above 80 ng / mL, preferably equal or above 100 ng / mL, irrespective of the GLP-1 -amide and GLP-l-Gly status, and / or the activity of PAM (SEQ ID No.: 1-10) is equal or above 14 Units / L, preferably equal or above 20 units / L, preferably equal or above 24 units / L, irrespective of a GLP-1 -amide and GLP-1 -Gly level. GLP-1 -Gly for use in treatment of a disease in a subject according to any of embodiments 1 to

[0434] 8, wherein GLP-1 -Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein GLP-1 -Gly is applied at a dosage of 0.1-500 pg / kg, preferably 1-400 pg / kg, more preferably 2-300 pg / mL, more preferably 2-300 pg / kg, more preferably 3-200 pg / kg, more preferably 5-100 pg / kg and wherein PAM is applied at a dosage of 1.5 - 54 Units / kg, preferably 1.7-33 Units / kg, more preferably 1.8-26 Units / kg, more preferably 1.9-20.6 Units / kg, more preferably 2-14 Units / kg. GLP-1 -Gly for use in treatment of a disease in a subject according to any of embodiments 1 to

[0435] 9, wherein GLP-1 -Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and in addition is to be used with Vitamin C and wherein Vitamin C is applied at a dosage of 1-10000 mg / kg, preferably 2-8000 mg / kg, more preferably 3-6000 mg / kg, more preferably 4-4000 mg / kg, more preferably 5-2000 mg / kg, more preferably 10-1000 mg / kg. GLP-1 -Gly for use in treatment of a disease in a subject according to any of embodiments 1 to

[0436] 10, wherein GLP-1 -Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) wherein said PAM is modified via amino acid manipulations, preferably via fusion to Albumins, e.g. serum Albumin or recombinant serum Albumin, more preferably via non covalent binding to serum Albumin due to a conjugated fatty acid chain to PAM, mor preferably via fusion with IgG Pc regions or Transferrin, more preferably modified to act as PEG-based carrier prodrug, most preferably via post-translational modifications attaching natural or synthetic polymers, whereas the natural or synthetic polymer to be used is HAP, preferably ELP, more preferably PAS, more preferably PSA, more preferably GLK, more preferably XTEN and most preferably PEG. GLP-1 -Gly for use in treatment of a disease in a subject according to any of embodiments 1 to

[0437] 11, wherein GLP-1 -Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) wherein said PAM is modified by pegylation. 13. Pharmaceutical combination or pharmaceutical composition comprising GLP-l-Gly and peptidylglycine a-amidating monooxygenase (PAM), wherein said combination may be a kits of part combination with at least two pharmaceutical compositions, wherein one of which comprises GLP-l-Gly and wherein the other comprises PAM or wherein said pharmaceutical composition comprises both GLP-l-Gly and peptidylglycine a-amidating monooxygenase (PAM), and wherein PAM and / or GLP-l-Gly may be in modified or unmodified form.

[0438] 14. Pharmaceutical combination or pharmaceutical composition according to embodiment 13, wherein said combination or composition comprises in addition Vitamin C.

[0439] 15. Pharmaceutical combination or pharmaceutical composition according to any of embodiment 13 or 14, wherein GLP-l-Gly is applied at a dosage of 0.1-500 pg / kg, preferably 1-400 pg / kg, more preferably 2-300 pg / mL, more preferably 2-300 pg / kg, more preferably 3-200 pg / kg, more preferably 5-100 pg / kg and wherein PAM is applied at a dosage of 1.5 - 54 Units / kg, preferably 1.7-33 Units / kg, more preferably 1.8-26 Units / kg, more preferably 1.9-20.6Units / kg, more preferably 2-14.0 Units / kg

[0440] 16. Pharmaceutical combination or pharmaceutical composition according to any of embodiments 13 or 15, wherein said combination or composition comprises in addition Vitamin C and wherein Vitamin C is applied at a dosage of 1-10000 mg / kg, preferably 2-8000 mg / kg, more preferably 3-6000 mg / kg, more preferably 4-4000 mg / kg, more preferably 5-2000 mg / kg, more preferably 10-1000 mg / kg.

[0441] 17. Pharmaceutical combination or pharmaceutical composition according to any of embodiment 13 or 16, wherein GLP-l-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) wherein said PAM is modified via amino acid manipulations, preferably via fusion to Albumins, e.g. serum Albumin or recombinant serum Albumin, more preferably via non covalent binding to serum Albumin due to a conjugated fatty acid chain to PAM, mor preferably via fusion with IgG Fc regions or Transferrin, preferably modified to act as PEG-based carrier prodrug, most preferably via post-translational modifications attaching natural or synthetic polymers, whereas the natural or synthetic polymer to be used is HAP, preferably ELP, more preferably PAS, more preferably PSA, more preferably GLK, more preferably XTEN and most preferably PEG.

[0442] The following embodiments further form part of the present invention:

[0443] 1. A method of treating a disease in a subject, said method comprising administering to said subject an effective amount of GLP-l-Gly, wherein GLP-l-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein PAM and / or GLP-l-Gly may be in modified or unmodified form. A method of treating a disease in a subject, said method comprising administering to said subject an effective amount of GLP-l-Gly according to embodiment 1, wherein GLP-l-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), wherein said subject is in need of elevated level of GLP-I-NH2 (GLP-1 -amide) in the circulation wherein elevated level means by at least 10 pg / mL, preferably by at least 20 pg / mL, more preferably by at least 30 pg / mL, more preferably by at least 40 pg / mL, most preferably by at least 50 pg / mL. A method of treating a disease in a subject, said method comprising administering to said subject an effective amount of GLP-l-Gly according to embodiment 1 or 2, wherein GLP-l-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), wherein said disease is characterized by a level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or a GLP-l-Gly / corresponding GLP-l-amide ratio above a threshold in a sample of bodily fluid of said subject and / or a GLP-l-amide level below a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or a GLP-l-amide level below a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof above a threshold and / or a GLP-l-amide level above a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or a GLP-l-amide level above a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof above a threshold. A method of treating a disease in a subject, said method comprising administering to said subject an effective amount of GLP-l-Gly according to any of embodiments 1 to 3, wherein GLP-l- Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), and wherein said disease is selected from the group comprising pulmonary hypertension and hypertension, type-2 diabetes, inflammatory bowel disease and / or Crohn's disease, sepsis and / or septic shock, cerebral infarction, mild cognitive impairment (MCI) and Alzheimer’s Disease (AD). A method of treating a disease in a subject, said method comprising administering to said subject an effective amount of GLP-l-Gly according to any of embodiments 1 to 4, wherein GLP-1- Gly is GLP-l-Gly (SEQ ID No.: 39-40), wherein GLP-l-Gly (SEQ ID No.: 39-40) is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), and wherein said disease is type-2 diabetes. A method of treating a disease in a subject, said method comprising administering to said subject an effective amount of GLP-l-Gly according to any of embodiments 1 to 5, wherein GLP-l- Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and in addition is to be used with Vitamin C. A method of treating a disease in a subject, said method comprising administering to said subject an effective amount of GLP-l-Gly according to any of embodiments 1 to 6, wherein GLP-l- Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein GLP-l-Gly and PAM are to be administered consecutively, or GLP-l-Gly and PAM are to be administered simultaneously. A method of treating a disease in a subject, said method comprising administering to said subject an effective amount of GLP-l-Gly according to any of embodiments 1 to 7, wherein the ratio of GLP-l-Gly and the corresponding GLP-l-amide in a bodily fluid is elevated over 1.5, preferably elevated over 1.6, more preferably elevated over 1.7, more preferably is elevated over 1.8, more preferable is elevated over 1.9, more preferably is higher than 2 and / or the concentration of PAM (SEQ ID No. 1-10) in a bodily fluid is equal or below 100 ng / mL (100 ng / mL to 0 ng / mL), preferably equal or below 90 ng / mL (90 ng / mL to 0 ng / mL), more preferably equal or below 80 ng / mL (80 ng / mL to 0 ng / mL), more preferably equal or below 60 ng / mL (60 ng / mL to 0 ng / mL), more preferably equal or below 50 ng / mL (50 ng / mL to 0 ng / mL) and most preferably equal or below 40 ng / mL (40 ng / mL to 0 ng / mL) and / or the activity of PAM (SEQ ID No. 1-10) in a bodily fluid is equal or below 24 Units / L, more preferably equal or below 20 Units / L, more preferably equal or below 16 Units / L, more preferably equal or below 14 Units / L, and most preferably equal or below 10 Units / L and / or GLP-1 -amide level is equal or below 20 pg / mL, preferably equal or below 15 pg / mL, preferably equal or below 10 pg / mL, preferably equal or below 5 pg / mL and / or

[0444] GLP-1 -Gly is equal or above 15 pg / mL, preferably equal or above 20 pg / mL, preferably equal or above 30 pg / mL, preferably is equal or above 50 pg / mL, and / or

[0445] GLP-1 -amide level is equal or above 70 pg / mL, irrespective of the status of PAM and / or GLP- 1-Gly, and / or the concentration of PAM (SEQ ID No.: 1-10) is equal or above 80 ng / mL, preferably equal or above 100 ng / mL, irrespective of the corresponding GLP-1 -amide and GLP-1 -Gly status, and / or the activity of PAM (SEQ ID No.: 1-10) is equal or above 14 Units / L, preferably equal or above 20 units / L, preferably equal or above 24 units / L, irrespective of a corresponding GLP-1 -amide and GLP-1 -Gly level.

[0446] 9. A method of treating a disease in a subject, said method comprising administering to said subject an effective amount of GLP-1 -Gly according to any of embodiments 1 to 8, wherein GLP-1 - Gly is applied at a dosage of 0.1-500 pg / kg, preferably 1-400 pg / kg, more preferably 2-300 pg / mL, more preferably 2-300 pg / kg, more preferably 3-200 pg / kg, more preferably 5-100 pg / kg and wherein PAM is applied at a dosage of 1.5 - 54 Units / kg, preferably 1.7-33 Units / kg, more preferably 1.8-26 Units / kg, more preferably 1.9-20.6Units / kg, more preferably 2-14.0 Units / kg.

[0447] 10. A method of treating a disease in a subject, said method comprising administering to said subject an effective amount of GLP-1 -Gly according to any of embodiments 1 to 9, wherein GLP-1 - Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and in addition is to be used with Vitamin C and wherein Vitamin C is applied at a dosage of 1- 10000 mg / kg, preferably 2-8000 mg / kg, more preferably 3-6000 mg / kg, more preferably 4-4000 mg / kg, more preferably 5-2000 mg / kg, more preferably 10-1000 mg / kg.

[0448] I L A method of treating a disease in a subject, said method comprising administering to said subject an effective amount of GLP-1 -Gly according to any of embodiments 1 to 10, wherein GLP-1 - Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) wherein said PAM is modified via amino acid manipulations, preferably via fusion to Albumins, e.g. serum Albumin or recombinant serum Albumin, more preferably via non covalent binding to serum Albumin due to a conjugated fatty acid chain to PAM, mor preferably via fusion with IgG Fc regions or Transferrin, preferably modified to act as PEG-based carrier prodrug, most preferably via post-translational modifications attaching natural or synthetic polymers, whereas the natural or synthetic polymer to be used is HAP, preferably ELP, more preferably PAS, more preferably PSA, more preferably GLK, more preferably XTEN and most preferably PEG.

[0449] 12. A method of treating a disease in a subject, said method comprising administering to said subject an effective amount of GLP-l-Gly according to any of embodiments 1 to 11, wherein GLP-1- Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) wherein said PAM is modified by pegylation.

[0450] It is apparent that the embodiments of the invention relating to the combinations, compositions and methods for treatment of certain diseases likewise relate to said combinations, compositions and methods for prevention of said diseases.

[0451] The following embodiments therefore also form also part of the present invention:

[0452] 1. Peptide-Gly for use in treatment and / or prevention of a disease in a subject, wherein Peptide- Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein PAM and / or Peptide-Gly may be in modified or unmodified form.

[0453] 2. Peptide-Gly for use in treatment and / or prevention of a disease in a subject according to embodiment 1, wherein Peptide-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM) and wherein Peptide-Gly is selected from the group comprising

[0454] • from the group of Adrenomedullin family of peptides: Adrenomedullin-2-Gly; Intermedin- Short-Gly and Proadrenomedullin N-20 terminal peptide-Gly,

[0455] • from the Bombesin / neurimedin-B / Ranatensin family of peptides: Gastrin-releasing peptide- Gly; Neuromedin-C-Gly; Neuromedin-B-Gly and Nouromedin-B-32-Gly,

[0456] • from the Calcitonin family of peptides: Calcitonin-Gly; Calcitonin gene-related peptide (CGRP) 1-Gly; CGRP 2-Gly and Islet amyloid polypeptide-Gly,

[0457] • from the Chromogranin / Secretogranin family of peptides: Chromogranin-A-Gly; ER-37-Gly; GR-44-Gly and Pancreastatin-Gly,

[0458] • from the FMRF-amide related peptide family of peptides: Neuropeptide AF-Gly; Neuropeptide FF-Gly; Neuropeptide SF-Gly Neuropeptide NPSF-Gly; Neuropeptide RFRP-l-Gly; Prolactinreleasing peptide PrRP20-Gly and Prolactin-releasing peptide PrRP31-Gly,

[0459] • from the Gastrin / Cholecystokinin family of peptides: Cholecystokinin -Gly; Cholecystokinin - 12-Gly; Cholecystokinin -18-Gly; Cholecystokinin -25-Gly; Cholecystokinin -33-Gly; Cholecystokinin -39-Gly; Cholecystokinin -5-Gly; Cholecystokinin -58-Gly; Cholecystokinin - 8-Gly; Cholecystokinin -7-Gly; Big-gastrin-Gly; Gastrin-Gly; Gastrin- 14-Gly; Gastrin-52-Gly; Gastrin-6-Gly and Gastrin-71-Gly,

[0460] • from the Glucagon family of peptides: Glucagon-like peptide (GLP) 1-Gly; GLP-l-Gly (7-37); Pituitary adenylate cyclase-activating polypeptide (PACP) 27-Gly; PACAP 38-Gly; Secretin- Gly; Somatoliberin-Gly; Intestinal Peptide PHM-27-Gly; Intestinal Peptide PHV-42-Gly and Vasoactive intestinal Peptide (VIP)-Gly,

[0461] • from the gonadotropin-releasing hormone family of peptides: Ginadoliberin- 1-Gly; Gonadoliberin-2-Gly; Progonadoliberin- 1-Gly and Progonadoliberin-2-Gly,

[0462] • from the Kisspeptin family of peptides: Kisspeptin-10-Gly; Kisspeptin-13-Gly; Kisspeptin- 14- Gly; Metastin-Gly and Metastasis-suppressor KISS- 1-Gly,

[0463] • from the Melanin-concentrating hormone family of peptides: Neuropeptide-glutamic acid- isoleucine-Gly and Pro-MCH-Gly,

[0464] • from the Neuromedin family of peptides: Neuromedin-S-Gly and Neuromedin-U-25-Gly,

[0465] • from the Neuropeptide-Y (NPY) family of peptides: Neuropeptide Y-Gly; Pancreatic hormone- Gly; Peptide YY-Gly and Peptide YY( 3-36)-Gly,

[0466] • from the opioid family of peptides: Deltrophin I-Gly,

[0467] • from the Orexin family of peptides: Orexin-A-Gly and Orexin-B-Gly,

[0468] • from the Proopiomelanocortin (POMC) family of peptides: Corticotropin-Gly; alpha- Melanotropin-Gly; gamma-Melanotropin-Gly and N-Terminal peptide of proopiomelanocortin- Gly (NPP-Gly),

[0469] • from the RFamide neuropeptide family of peptides: Orexigenic neuropeptide QRF-Gly (QRF- Gly),

[0470] • from the Sauvagine / corticotropin-releasing factor / urotensin I family of peptides: Urocortin-Gly and Urocortin-3-Gly,

[0471] • from the Tachykinin family of peptides: Substance P-Gly (RPKPQQFFGLMG) and Substance P-Gly (RPKPQFFGLMG); Neurokinin A-Gly; Neuropeptide K-Gly and Neurokinin-B-Gly,

[0472] • from the Thyrotropin Releasing Hormone (TRH) family of peptides: pro-TRH-Gly and TRH- Gly,

[0473] • from the Vasopressin / Oxytocin family of peptides: Oxytocin-Gly and Arg-Vasopressin-Gly,

[0474] • from the VGF nerve growth factor inducible protein (VGF) family of peptides: Antimicrobial peptide VGF-Gly (aa554-578) and Neurosecretory protein VGF-Gly.

[0475] 3. Peptide-Gly for use in treatment and / or prevention of a disease in a subject according to embodiments 1 or 2, wherein Peptide-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM) and wherein Peptide-Gly is Adrenomedullin (ADM)-Gly and wherein ADM-Gly may be in modified or unmodified form. Peptide-Gly for use in treatment and / or prevention of a disease in a subject according to embodiments 1 to 3, wherein Peptide-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM), wherein said subject is in need of elevated level of Peptide- NH2 (Peptide-amide) in the circulation wherein elevated level means by at least 10 pg / mL, preferably by at least 20 pg / mL, more preferably by at least 30 pg / mL, more preferably by at least 40 pg / mL, most preferably by at least 50 pg / mL. Peptide-Gly for use in treatment and / or prevention of a disease in a subject according to embodiments 1 to 4, wherein Peptide-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM), wherein said disease is characterized by a level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or a Peptide-Gly / corresponding Peptide-amide ratio above a threshold in a sample of bodily fluid of said subject and / or a Peptide- amide level below a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or a Peptide- amide level below a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof above a threshold and / or a Peptide- amide level above a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or a Peptide- amide level above a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof above a threshold. Peptide-Gly for use in treatment and / or prevention of a disease in a subject according to any of embodiments 1 to 5, wherein Peptide-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM), and wherein said disease is selected from the group comprising pulmonary hypertension and hypertension, type-2 diabetes, inflammatory bowel disease and / or Crohn's disease, sepsis and / or septic shock, cerebral infarction, mild cognitive impairment (MCI) and Alzheimer’s Disease (AD). Peptide-Gly for use in treatment and / or prevention of a disease in a subject according to any of the preceding embodiments, wherein Peptide-Gly is GLP-l-Gly (SEQ ID No.: 39-40), wherein GLP-l-Gly (SEQ ID No.: 39-40) is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), and wherein said disease is type-2 diabetes. Peptide-Gly for use in treatment and / or prevention of a disease in a subject according to any of embodiments 1 to 7, wherein Peptide-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM) and in addition is to be used with Vitamin C. Peptide-Gly for use in treatment and / or prevention of a disease in a subject according to any of embodiments 1 to 8, wherein Peptide-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM) and wherein Peptide-Gly and PAM are to be administered consecutively, or Peptide-Gly and PAM are to be administered simultaneously. Peptide-Gly for use in treatment and / or prevention of a disease in a subject according to any of embodiments 1 to 9, wherein Peptide-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM) and wherein the ratio of Peptide-Gly and the corresponding Peptide-amide in a bodily fluid is elevated over 1.5, preferably elevated over 1.6, more preferably elevated over 1.7, more preferably is elevated over 1.8, more preferable is elevated over 1.9, more preferably is higher than 2 and / or the concentration of PAM (SEQ ID No. 1-10) in a bodily fluid is equal or below 100 ng / mL (100 ng / mL to 0 ng / mL), preferably equal or below 90 ng / mL (90 ng / mL to 0 ng / mL), more preferably equal or below 80 ng / mL (80 ng / mL to 0 ng / mL), more preferably equal or below 60 ng / mL (60 ng / mL to 0 ng / mL), more preferably equal or below 50 ng / mL (50 ng / mL to 0 ng / mL) and most preferably equal or below 40 ng / mL (40 ng / mL to 0 ng / mL)and / or the activity of PAM (SEQ ID No. 1-10) in a bodily fluid is equal or below 24 Units / L, more preferably equal or below 20 Units / L, more preferably equal or below 16 Units / L, more preferably equal or below 14 Units / L, and most preferably equal or below 10 Units / L and / or

[0476] Peptide-amide level is equal or below 20 pg / mL, preferably equal or below 15 pg / mL, preferably equal or below 10 pg / mL, preferably equal or below 5 pg / mL and / or

[0477] Peptide- is equal or above 15 pg / mL, preferably equal or above 20 pg / mL, preferably equal or above 30 pg / mL, preferably is equal or above 50 pg / mL, and / or

[0478] Peptide-amide level is equal or above 70 pg / mL, irrespective of the status of PAM and / or Peptide-Gly, and / or the concentration of PAM (SEQ ID No.: 1-10) is equal or above 80 ng / mL, preferably equal or above 100 ng / mL, irrespective of the corresponding Peptide-amide and Peptide-Gly status, and / or the activity of PAM (SEQ ID No.: 1-10) is equal or above 14 Units / L, preferably equal or above 20 units / L, preferably equal or above 24 units / L, irrespective of a corresponding Peptide-amide and Peptide-Gly level. Peptide-Gly for use in treatment and / or prevention of a disease in a subject according to any of embodiments 1 to 10, wherein Peptide-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM) and wherein Peptide-Gly is applied at a dosage of 0.1-500 pg / kg, preferably 1-400 pg / kg, more preferably 2-300 pg / mL, more preferably 2-300 pg / kg, more preferably 3-200 pg / kg, more preferably 5-100 pg / kg and wherein PAM is applied at a dosage of 1.5 - 54 Units / kg, preferably 1.7-33 Units / kg, more preferably 1.8-26 Units / kg, more preferably 1.9-20.6 Units / kg, more preferably 2-14 Units / kg. Peptide-Gly for use in treatment and / or prevention of a disease in a subject according to any of embodiments 1 to 11, wherein Peptide-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM) and in addition is to be used with Vitamin C and wherein Vitamin C is applied at a dosage of 1-10000 mg / kg, preferably 2-8000 mg / kg, more preferably 3-6000 mg / kg, more preferably 4-4000 mg / kg, more preferably 5-2000 mg / kg, more preferably 10-1000 mg / kg. Peptide-Gly for use in treatment and / or prevention of a disease in a subject according to any of embodiments 1 to 12, wherein Peptide-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM) wherein said PAM is modified via amino acid manipulations, preferably via fusion to Albumins, e.g. serum Albumin or recombinant serum Albumin, more preferably via non covalent binding to serum Albumin due to a conjugated fatty acid chain to PAM, mor preferably via fusion with IgG Fc regions or Transferrin, more preferably modified to act as PEG-based carrier prodrug, most preferably via post-translational modifications attaching natural or synthetic polymers, whereas the natural or synthetic polymer to be used is HAP, preferably ELP, more preferably PAS, more preferably PSA, more preferably GLK, more preferably XTEN and most preferably PEG. Peptide-Gly for use in treatment and / or prevention of a disease in a subject according to any of embodiments 1 to 13, wherein Peptide-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM) wherein said PAM is modified by pegylation. Pharmaceutical combination or pharmaceutical composition comprising Peptide-Gly and peptidylglycine a-amidating monooxygenase (PAM), wherein said combination may be a kits of part combination with at least two pharmaceutical compositions, wherein one of which comprises Peptide-Gly and wherein the other comprises PAM or wherein said pharmaceutical composition comprises both Peptide-Gly and peptidylglycine a-amidating monooxygenase (PAM), and wherein PAM and / or Peptide-Gly may be in modified or unmodified form. Pharmaceutical combination or pharmaceutical composition according to embodiment 15, wherein said combination or composition comprises in addition Vitamin C. Pharmaceutical combination or pharmaceutical composition according to any of embodiment 15 or 16, wherein Peptide-Gly is applied at a dosage of 0.1-500 pg / kg, preferably 1-400 pg / kg, more preferably 2-300 pg / mL, more preferably 2-300 pg / kg, more preferably 3-200 pg / kg, more preferably 5-100 pg / kg and wherein PAM is applied at a dosage of 1.5 - 54 Units / kg, preferably 1.7-33 Units / kg, more preferably 1.8-26 Units / kg, more preferably 1.9-20.6Units / kg, more preferably 2-14.0 Units / kg. Pharmaceutical combination or pharmaceutical composition according to any of embodiments 15 or 17, wherein said combination or composition comprises in addition Vitamin C and wherein Vitamin C is applied at a dosage of 1-10000 mg / kg, preferably 2-8000 mg / kg, more preferably 3-6000 mg / kg, more preferably 4-4000 mg / kg, more preferably 5-2000 mg / kg, more preferably 10-1000 mg / kg. Pharmaceutical combination or pharmaceutical composition according to any of embodiment 15 or 18, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) wherein said PAM is modified via amino acid manipulations, preferably via fusion to Albumins, e.g. serum Albumin or recombinant serum Albumin, more preferably via non covalent binding to serum Albumin due to a conjugated fatty acid chain to PAM, mor preferably via fusion with IgG Fc regions or Transferrin, preferably modified to act as PEG-based carrier prodrug, most preferably via post-translational modifications attaching natural or synthetic polymers, whereas the natural or synthetic polymer to be used is HAP, preferably EEP, more preferably PAS, more preferably PSA, more preferably GLK, more preferably XTEN and most preferably PEG. 20. A method of treating and / or preventing a disease in a subject, said method comprising administering to said subject an effective amount of Peptide-Gly, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein PAM and / or Peptide-Gly may be in modified or unmodified form.

[0479] 21. A method of treating and / or preventing a disease in a subject, said method comprising administering to said subject an effective amount of Peptide-Gly according to embodiment 20, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein Peptide-Gly is selected from the group comprising

[0480] • from the group of Adrenomedullin family of peptides: Adrenomedullin-2-Gly; Intermedin- Short-Gly and Proadrenomedullin N-20 terminal peptide-Gly,

[0481] • from the Bombesin / neurimedin-B / Ranatensin family of peptides: Gastrin-releasing peptide- Gly; Neuromedin-C-Gly; Neuromedin-B-Gly and Nouromedin-B-32-Gly,

[0482] • from the Calcitonin family of peptides: Calcitonin-Gly; Calcitonin gene-related peptide (CGRP) 1-Gly; CGRP 2-Gly and Islet amyloid polypeptide-Gly,

[0483] • from the Chromogranin / Secretogranin family of peptides: Chromogranin-A-Gly; ER-37-Gly; GR-44-Gly and Pancreastatin-Gly,

[0484] • from the FMRF-amide related peptide family of peptides: Neuropeptide AF-Gly; Neuropeptide FF-Gly; Neuropeptide SF-Gly Neuropeptide NPSF-Gly; Neuropeptide RFRP-l-Gly; Prolactinreleasing peptide PrRP20-Gly and Prolactin-releasing peptide PrRP31-Gly,

[0485] • from the Gastrin / Cholecystokinin family of peptides: Cholecystokinin-Gly; Cholecystokinin-

[0486] 12-Gly; Cholecystokinin- 18-Gly; Cholecystokinin-25 -Gly; Cholecystokinin-33 -Gly;

[0487] Cholecystokinin-39-Gly; Cholecystokinin-5 -Gly; Cholecystokinin-58-Gly; Cholecystokinin-8- Gly; Cholecystokinin-7-Gly; Big-gastrin-Gly; Gastrin-Gly; Gastrin- 14-Gly; Gastrin-52-Gly; Gastrin-6-Gly and Gastrin-71 -Gly,

[0488] • from the Glucagon family of peptides: Glucagon-like peptide (GLP) 1-Gly; GLP-l-Gly (7-37); Pituitary adenylate cyclase-activating polypeptide (PACP) 27-Gly; PACAP 38-Gly; Secretin- Gly; Somatoliberin-Gly; Intestinal Peptide PHM-27-Gly; Intestinal Peptide PHV-42-Gly and Vasoactive intestinal Peptide (VIP)-Gly,

[0489] • from the gonadotropin-releasing hormone family of peptides: Ginadoliberin- 1-Gly; Gonadoliberin-2-Gly; Progonadoliberin- 1-Gly and Progonadoliberin-2-Gly,

[0490] • from the Kisspeptin family of peptides: Kisspeptin-10-Gly; Kisspeptin-13-Gly; Kisspeptin- 14- Gly; Metastin-Gly and Metastasis-suppressor KISS- 1-Gly,

[0491] • from the Melanin-concentrating hormone family of peptides: Neuropeptide-glutamic acid- isoleucine-Gly and Pro-MCH-Gly, from the Neuromedin family of peptides: Neuromedin-S-Gly and Neuromedin-U-25-Gly, • from the Neuropeptide-Y (NPY) family of peptides: Neuropeptide Y-Gly; Pancreatic hormone- Gly; Peptide YY-Gly and Peptide YY( 3-36)-Gly,

[0492] • from the opioid family of peptides: Deltrophin I-Gly,

[0493] • from the Orexin family of peptides: Orexin-A-Gly and Orexin-B-Gly,

[0494] • from the Proopiomelanocortin (POMC) family of peptides: Corticotropin-Gly; alpha- Melanotropin-Gly; gamma-Melanotropin-Gly and N-Terminal peptide of proopiomelanocortin- Gly (NPP-Gly),

[0495] • from the RFamide neuropeptide family of peptides: Orexigenic neuropeptide QRF-Gly (QRF- Gly),

[0496] • from the Sauvagine / corticotropin-releasing factor / urotensin I family of peptides: Urocortin-Gly and Urocortin-3-Gly,

[0497] • from the Tachykinin family of peptides: Substance P-Gly (RPKPQQFFGLMG) and Substance P-Gly (RPKPQFFGLMG); Neurokinin A-Gly; Neuropeptide K-Gly and Neurokinin-B-Gly,

[0498] • from the Thyrotropin Releasing Hormone (TRH) family of peptides: pro-TRH-Gly and TRH- Gly,

[0499] • from the Vasopressin / Oxytocin family of peptides: Oxytocin-Gly and Arg-Vasopressin-Gly,

[0500] • from the VGF nerve growth factor inducible protein (VGF) family of peptides: Antimicrobial peptide VGF-Gly (aa554-578) and Neurosecretory protein VGF-Gly.

[0501] 22. A method of treating and / or preventing a disease in a subject, said method comprising administering to said subject an effective amount of Peptide-Gly according to embodiments 20 or 21, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein Peptide Gly is ADM-Gly and wherein ADM-Gly may be in modified or unmodified form.

[0502] 23. A method of treating and / or preventing a disease in a subject, said method comprising administering to said subject an effective amount of Peptide-Gly according to embodiment 20 to 22, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), wherein said subject is in need of elevated level of Peptide-fMT (Peptide-amide) in the circulation wherein elevated level means by at least 10 pg / mL, preferably by at least 20 pg / mL, more preferably by at least 30 pg / mL, more preferably by at least 40 pg / mL, most preferably by at least 50 pg / mL.

[0503] 24. A method of treating and / or preventing a disease in a subject, said method comprising administering to said subject an effective amount of Peptide-Gly according to embodiment 20 to 23, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), wherein said disease is characterized by a level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or a Peptide-Gly / corresponding Peptide-amide ratio above a threshold in a sample of bodily fluid of said subject and / or a Peptide-amide level below a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or a Peptide-amide level below a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof above a threshold and / or a Peptide-amide level above a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or a Peptide-amide level above a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof above a threshold. A method of treating and / or preventing a disease in a subject, said method comprising administering to said subject an effective amount of Peptide-Gly according to any of embodiments 20 to 24, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), and wherein said disease is selected from the group comprising pulmonary hypertension and hypertension, type-2 diabetes, inflammatory bowel disease and / or Crohn's disease, sepsis and / or septic shock, cerebral infarction, mild cognitive impairment (MCI) and Alzheimer’s Disease (AD). A method of treating and / or preventing a disease in a subject, said method comprising administering to said subject an effective amount of Peptide-Gly according to any of embodiments 20 to 25, wherein Peptide-Gly is GLP-l-Gly (SEQ ID No.: 39-40), wherein GLP- 1-Gly (SEQ ID No.: 39-40) is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), and wherein said disease is type-2 diabetes. A method of treating and / or preventing a disease in a subject, said method comprising administering to said subject an effective amount of Peptide-Gly according to any of embodiments 20 to 26, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and in addition is to be used with Vitamin C. A method of treating and / or preventing a disease in a subject, said method comprising administering to said subject an effective amount of Peptide-Gly according to any of embodiments 20 to 27, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein Peptide-Gly and PAM are to be administered consecutively, or Peptide-Gly and PAM are to be administered simultaneously. A method of treating and / or preventing a disease in a subject, said method comprising administering to said subject an effective amount of Peptide-Gly according to any of embodiments 20 to 28, wherein the ratio of Peptide-Gly and the corresponding Peptide-amide in a bodily fluid is elevated over 1.5, preferably elevated over 1.6, more preferably elevated over 1.7, more preferably is elevated over 1.8, more preferable is elevated over 1.9, more preferably is higher than 2 and / or the concentration of PAM (SEQ ID No. 1-10) in a bodily fluid is equal or below 100 ng / mL (100 ng / mL to 0 ng / mL), preferably equal or below 90 ng / mL (90 ng / mL to 0 ng / mL), more preferably equal or below 80 ng / mL (80 ng / mL to 0 ng / mL), more preferably equal or below 60 ng / mL (60 ng / mL to 0 ng / mL), more preferably equal or below 50 ng / mL (50 ng / mL to 0 ng / mL) and most preferably equal or below 40 ng / mL (40 ng / mL to 0 ng / mL) and / or the activity of PAM (SEQ ID No. 1-10) in a bodily fluid is equal or below 24 Units / L, more preferably equal or below 20 Units / L, more preferably equal or below 16 Units / L, more preferably equal or below 14 Units / L, and most preferably equal or below 10 Units / L and / or

[0504] Peptide-amide level is equal or below 20 pg / mL, preferably equal or below 15 pg / mL, preferably equal or below 10 pg / mL, preferably equal or below 5 pg / mL and / or

[0505] Peptide-Gly is equal or above 15 pg / mL, preferably equal or above 20 pg / mL, preferably equal or above 30 pg / mL, preferably is equal or above 50 pg / mL, and / or

[0506] Peptide-amide level is equal or above 70 pg / mL, irrespective of the status of PAM and / or Peptide-Gly, and / or the concentration of PAM (SEQ ID No.: 1-10) is equal or above 80 ng / mL, preferably equal or above 100 ng / mL, irrespective of the corresponding Peptide-amide and Peptide-Gly status, and / or the activity of PAM (SEQ ID No.: 1-10) is equal or above 14 Units / L, preferably equal or above 20 units / L, preferably equal or above 24 units / L, irrespective of a corresponding Peptide-amide and Peptide-Gly level. A method of treating and / or preventing a disease in a subject, said method comprising administering to said subject an effective amount of Peptide-Gly according to any of embodiments 20 to 29, wherein Peptide-Gly is applied at a dosage of 0.1-500 pg / kg, preferably 1-400 pg / kg, more preferably 2-300 pg / mL, more preferably 2-300 pg / kg, more preferably 3- 200 pg / kg, more preferably 5-100 pg / kg and wherein PAM is applied at a dosage of 1.5 - 54 Units / kg, preferably 1.7-33 Units / kg, more preferably 1.8-26 Units / kg, more preferably 1.9- 20.6Units / kg, more preferably 2-14.0 Units / kg. A method of treating and / or preventing a disease in a subject, said method comprising administering to said subject an effective amount of Peptide-Gly according to any of embodiments 20 to 30, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and in addition is to be used with Vitamin C and wherein Vitamin C is applied at a dosage of 1-10000 mg / kg, preferably 2-8000 mg / kg, more preferably 3-6000 mg / kg, more preferably 4-4000 mg / kg, more preferably 5-2000 mg / kg, more preferably 10-1000 mg / kg. A method of treating and / or preventing a disease in a subject, said method comprising administering to said subject an effective amount of Peptide-Gly according to any of embodiments 20 to 31, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) wherein said PAM is modified via amino acid manipulations, preferably via fusion to Albumins, e.g. serum Albumin or recombinant serum Albumin, more preferably via non covalent binding to serum Albumin due to a conjugated fatty acid chain to PAM, mor preferably via fusion with IgG Fc regions or Transferrin, preferably modified to act as PEG-based carrier prodrug, most preferably via post-translational modifications attaching natural or synthetic polymers, whereas the natural or synthetic polymer to be used is HAP, preferably ELP, more preferably PAS, more preferably PSA, more preferably GLK, more preferably XTEN and most preferably PEG. A method of treating and / or preventing a disease in a subject, said method comprising administering to said subject an effective amount of Peptide-Gly according to any of embodiments 20 to 32, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) wherein said PAM is modified by pegylation. Moreover, the following embodiments therefore also form part of the present invention:

[0507] 1. ADM-Gly for use in treatment and / or prevention of a disease in a subject, wherein ADM-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein PAM and / or ADM-Gly may be in modified or unmodified form.

[0508] 2. ADM-Gly for use in treatment and / or prevention of a disease in a subject according to embodiment 1, wherein ADM-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM), wherein said subject is in need of an elevated level of ADM- NH2 (ADM-amide) in the circulation wherein elevated level means by at least 10 pg / mL, preferably by at least 20 pg / mL, more preferably by at least 30 pg / mL, more preferably by at least 40 pg / mL, most preferably by at least 50 pg / mL.

[0509] 3. ADM-Gly for use in treatment and / or prevention of a disease in a subject according to embodiments 1 or 2, wherein ADM-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM), wherein said disease is characterized by a level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or an ADM-Gly / ADM-amide ratio above a threshold in a sample of bodily fluid of said subject and / or an ADM- amide level below a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or an ADM- amide level below a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof above a threshold and / or an ADM- amide level above a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or an ADM- amide level above a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof above a threshold.

[0510] 4. ADM-Gly for use in treatment and / or prevention of a disease in a subject according to any of embodiments 1 to 3, wherein ADM-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM), and wherein said disease is selected from the group comprising pulmonary hypertension and hypertension, type-2 diabetes, inflammatory bowel disease and / or Crohn's disease, sepsis and / or septic shock, cerebral infarction, mild cognitive impairment (MCI) and Alzheimer’s Disease (AD). ADM-Gly for use in treatment and / or prevention of a disease in a subject according to any of embodiments 1 to 4, wherein ADM-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM) and in addition is to be used with Vitamin C. ADM-Gly for use in treatment and / or prevention of a disease in a subject according to any of embodiments 1 to 5, wherein ADM-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM) and wherein ADM-Gly and PAM are to be administered consecutively, or ADM-Gly and PAM are to be administered simultaneously. ADM-Gly for use in treatment and / or prevention of a disease in a subject according to any of embodiments 1 to 6, wherein ADM-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM) and wherein the ratio of ADM-Gly and the ADM-amide in a bodily fluid is elevated over 1.5, preferably elevated over 1.6, more preferably elevated over 1.7, more preferably is elevated over 1.8, more preferable is elevated over 1.9, more preferably is higher than 2 and / or the concentration of PAM (SEQ ID No. 1-10) in a bodily fluid is equal or below 100 ng / mL (100 ng / mL to 0 ng / mL), preferably equal or below 90 ng / mL (90 ng / mL to 0 ng / mL), more preferably equal or below 80 ng / mL (80 ng / mL to 0 ng / mL), more preferably equal or below 60 ng / mL (60 ng / mL to 0 ng / mL), more preferably equal or below 50 ng / mL (50 ng / mL to 0 ng / mL) and most preferably equal or below 40 ng / mL (40 ng / mL to 0 ng / mL)and / or the activity of PAM (SEQ ID No. 1-10) in a bodily fluid is equal or below 24 Units / L, more preferably equal or below 20 Units / L, more preferably equal or below 16 Units / L, more preferably equal or below 14 Units / L, and most preferably equal or below 10 Units / L and / or

[0511] ADM-amide level is equal or below 20 pg / mL, preferably equal or below 15 pg / mL, preferably equal or below 10 pg / mL, preferably equal or below 5 pg / mL and / or

[0512] ADM- is equal or above 15 pg / mL, preferably equal or above 20 pg / mL, preferably equal or above 30 pg / mL, preferably is equal or above 50 pg / mL, and / or

[0513] ADM-amide level is equal or above 70 pg / mL, irrespective of the status of PAM and / or ADM- Gly, and / or the concentration of PAM (SEQ ID No.: 1-10) is equal or above 80 ng / mL, preferably equal or above 100 ng / mL, irrespective of the ADM-amide and ADM-Gly status, and / or the activity of PAM (SEQ ID No.: 1-10) is equal or above 14 Units / L, preferably equal or above 20 units / L, preferably equal or above 24 units / L, irrespective of a ADM-amide and ADM-Gly level. ADM-Gly for use in treatment and / or prevention of a disease in a subject according to any of embodiments 1 to 7, wherein ADM-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM) and wherein ADM-Gly is applied at a dosage of 0.1-500 pg / kg, preferably 1-400 pg / kg, more preferably 2-300 pg / mL, more preferably 2-300 pg / kg, more preferably 3-200 pg / kg, more preferably 5-100 pg / kg and wherein PAM is applied at a dosage of 1.5 - 54 Units / kg, preferably 1.7-33 Units / kg, more preferably 1.8-26 Units / kg, more preferably 1.9-20.6 Units / kg, more preferably 2-14 Units / kg. ADM-Gly for use in treatment and / or prevention of a disease in a subject according to any of embodiments 1 to 8, wherein ADM-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM) and in addition is to be used with Vitamin C and wherein Vitamin C is applied at a dosage of 1-10000 mg / kg, preferably 2-8000 mg / kg, more preferably 3-6000 mg / kg, more preferably 4-4000 mg / kg, more preferably 5-2000 mg / kg, more preferably 10-1000 mg / kg. ADM-Gly for use in treatment and / or prevention of a disease in a subject according to any of embodiments 1 to 9, wherein ADM-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM) wherein said PAM is modified via amino acid manipulations, preferably via fusion to Albumins, e.g. serum Albumin or recombinant serum Albumin, more preferably via non covalent binding to serum Albumin due to a conjugated fatty acid chain to PAM, mor preferably via fusion with IgG Fc regions or Transferrin, more preferably modified to act as PEG-based carrier prodrug, most preferably via post-translational modifications attaching natural or synthetic polymers, whereas the natural or synthetic polymer to be used is HAP, preferably ELP, more preferably PAS, more preferably PSA, more preferably GLK, more preferably XTEN and most preferably PEG. ADM-Gly for use in treatment and / or prevention of a disease in a subject according to any of embodiments 1 to 10, wherein ADM-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM) wherein said PAM is modified by pegylation. Pharmaceutical combination or pharmaceutical composition comprising ADM-Gly and peptidylglycine a-amidating monooxygenase (PAM), wherein said combination may be a kits of part combination with at least two pharmaceutical compositions, wherein one of which comprises ADM-Gly and wherein the other comprises PAM or wherein said pharmaceutical composition comprises both ADM-Gly and peptidylglycine a-amidating monooxygenase (PAM), and wherein PAM and / or ADM-Gly may be in modified or unmodified form. Pharmaceutical combination or pharmaceutical composition according to embodiment 12, wherein said combination or composition comprises in addition Vitamin C. Pharmaceutical combination or pharmaceutical composition according to any of embodiment 12 or 13, wherein ADM-Gly is applied at a dosage of 0.1-500 pg / kg, preferably 1-400 pg / kg, more preferably 2-300 pg / mL, more preferably 2-300 pg / kg, more preferably 3-200 pg / kg, more preferably 5-100 pg / kg and wherein PAM is applied at a dosage of 1.5 - 54 Units / kg, preferably 1.7-33 Units / kg, more preferably 1.8-26 Units / kg, more preferably 1.9-20.6Units / kg, more preferably 2-14.0 Units / kg Pharmaceutical combination or pharmaceutical composition according to any of embodiments 12 or 14, wherein said combination or composition comprises in addition Vitamin C and wherein Vitamin C is applied at a dosage of 1-10000 mg / kg, preferably 2-8000 mg / kg, more preferably 3-6000 mg / kg, more preferably 4-4000 mg / kg, more preferably 5-2000 mg / kg, more preferably 10-1000 mg / kg. Pharmaceutical combination or pharmaceutical composition according to any of embodiment 12 or 15, wherein ADM-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) wherein said PAM is modified via amino acid manipulations, preferably via fusion to Albumins, e.g. serum Albumin or recombinant serum Albumin, more preferably via non covalent binding to serum Albumin due to a conjugated fatty acid chain to PAM, mor preferably via fusion with IgG Fc regions or Transferrin, preferably modified to act as PEG-based carrier prodrug, most preferably via post-translational modifications attaching natural or synthetic polymers, whereas the natural or synthetic polymer to be used is HAP, preferably EEP, more preferably PAS, more preferably PSA, more preferably GLK, more preferably XTEN and most preferably PEG. The following embodiments therefore further form part of the present invention:

[0514] 1. A method of treating and / or preventing a disease in a subject, said method comprising administering to said subject an effective amount of ADM-Gly, wherein ADM-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein PAM and / or ADM-Gly may be in modified or unmodified form.

[0515] 2. A method of treating and / or preventing a disease in a subject, said method comprising administering to said subject an effective amount of ADM-Gly according to embodiment 1, wherein ADM-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), wherein said subject is in need of elevated level of ADM-NH2 (ADM- amide) in the circulation wherein elevated level means by at least 10 pg / mL, preferably by at least 20 pg / mL, more preferably by at least 30 pg / mL, more preferably by at least 40 pg / mL, most preferably by at least 50 pg / mL.

[0516] 3. A method of treating and / or preventing a disease in a subject, said method comprising administering to said subject an effective amount of ADM-Gly according to embodiment 1 or 2, wherein ADM-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), wherein said disease is characterized by a level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or an ADM-Gly / corresponding ADM-amide ratio above a threshold in a sample of bodily fluid of said subject and / or an ADM-amide level below a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or an ADM-amide level below a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof above a threshold and / or an ADM-amide level above a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or an ADM-amide level above a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof above a threshold. A method of treating and / or preventing a disease in a subject, said method comprising administering to said subject an effective amount of ADM-Gly according to any of embodiments 1 to 3, wherein ADM-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM), and wherein said disease is selected from the group comprising pulmonary hypertension and hypertension, type-2 diabetes, inflammatory bowel disease and / or Crohn's disease, sepsis and / or septic shock, cerebral infarction, mild cognitive impairment (MCI) and Alzheimer’s Disease (AD). A method of treating and / or preventing a disease in a subject, said method comprising administering to said subject an effective amount of ADM-Gly according to any of embodiments 1 to 4, wherein ADM-Gly is GLP-l-Gly (SEQ ID No.: 39-40), wherein GLP-1- Gly (SEQ ID No.: 39-40) is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), and wherein said disease is type-2 diabetes. A method of treating and / or preventing a disease in a subject, said method comprising administering to said subject an effective amount of ADM-Gly according to any of embodiments 1 to 5, wherein ADM-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM) and in addition is to be used with Vitamin C. A method of treating and / or preventing a disease in a subject, said method comprising administering to said subject an effective amount of ADM-Gly according to any of embodiments 1 to 6, wherein ADM-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM) and wherein ADM-Gly and PAM are to be administered consecutively, or ADM-Gly and PAM are to be administered simultaneously. A method of treating and / or preventing a disease in a subject, said method comprising administering to said subject an effective amount of ADM-Gly according to any of embodiments 1 to 7, wherein the ratio of ADM-Gly and the corresponding ADM-amide in a bodily fluid is elevated over 1.5, preferably elevated over 1.6, more preferably elevated over 1.7, more preferably is elevated over 1.8, more preferable is elevated over 1.9, more preferably is higher than 2 and / or the concentration of PAM (SEQ ID No. 1-10) in a bodily fluid is equal or below 100 ng / mL (100 ng / mL to 0 ng / mL), preferably equal or below 90 ng / mL (90 ng / mL to 0 ng / mL), more preferably equal or below 80 ng / mL (80 ng / mL to 0 ng / mL), more preferably equal or below 60 ng / mL (60 ng / mL to 0 ng / mL), more preferably equal or below 50 ng / mL (50 ng / mL to 0 ng / mL) and most preferably equal or below 40 ng / mL (40 ng / mL to 0 ng / mL) and / or the activity of PAM (SEQ ID No. 1-10) in a bodily fluid is equal or below 24 Units / L, more preferably equal or below 20 Units / L, more preferably equal or below 16 Units / L, more preferably equal or below 14 Units / L, and most preferably equal or below 10 Units / L and / or

[0517] ADM-amide level is equal or below 20 pg / mL, preferably equal or below 15 pg / mL, preferably equal or below 10 pg / mL, preferably equal or below 5 pg / mL and / or

[0518] ADM-Gly is equal or above 15 pg / mL, preferably equal or above 20 pg / mL, preferably equal or above 30 pg / mL, preferably is equal or above 50 pg / mL, and / or

[0519] ADM-amide level is equal or above 70 pg / mL, irrespective of the status of PAM and / or ADM- Gly, and / or the concentration of PAM (SEQ ID No.: 1-10) is equal or above 80 ng / mL, preferably equal or above 100 ng / mL, irrespective of the corresponding ADM-amide and ADM-Gly status, and / or the activity of PAM (SEQ ID No.: 1-10) is equal or above 14 Units / L, preferably equal or above 20 units / L, preferably equal or above 24 units / L, irrespective of a corresponding ADM-amide and ADM-Gly level. A method of treating and / or preventing a disease in a subject, said method comprising administering to said subject an effective amount of ADM-Gly according to any of embodiments 1 to 8, wherein ADM-Gly is applied at a dosage of 0.1-500 pg / kg, preferably 1- 400 pg / kg, more preferably 2-300 pg / mL, more preferably 2-300 pg / kg, more preferably 3-200 pg / kg, more preferably 5-100 pg / kg and wherein PAM is applied at a dosage of 1.5 - 54 Units / kg, preferably 1.7-33 Units / kg, more preferably 1.8-26 Units / kg, more preferably 1.9- 20.6Units / kg, more preferably 2-14.0 Units / kg. A method of treating and / or preventing a disease in a subject, said method comprising administering to said subject an effective amount of ADM-Gly according to any of embodiments 1 to 9, wherein ADM-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM) and in addition is to be used with Vitamin C and wherein Vitamin C is applied at a dosage of 1-10000 mg / kg, preferably 2-8000 mg / kg, more preferably 3-6000 mg / kg, more preferably 4-4000 mg / kg, more preferably 5-2000 mg / kg, more preferably 10-1000 mg / kg. 11. A method of treating and / or preventing a disease in a subject, said method comprising administering to said subject an effective amount of ADM-Gly according to any of embodiments 1 to 10, wherein ADM-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM) wherein said PAM is modified via amino acid manipulations, preferably via fusion to Albumins, e.g. serum Albumin or recombinant serum Albumin, more preferably via non covalent binding to serum Albumin due to a conjugated fatty acid chain to PAM, mor preferably via fusion with IgG Fc regions or Transferrin, preferably modified to act as PEG-based carrier prodrug, most preferably via post-translational modifications attaching natural or synthetic polymers, whereas the natural or synthetic polymer to be used is HAP, preferably ELP, more preferably PAS, more preferably PSA, more preferably GLK, more preferably XTEN and most preferably PEG.

[0520] 12. A method of treating and / or preventing a disease in a subject, said method comprising administering to said subject an effective amount of ADM-Gly according to any of embodiments 1 to 11, wherein ADM-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM) wherein said PAM is modified by pegylation.

[0521] The following embodiments therefore form also part of the present invention:

[0522] 1. GLP-l-Gly for use in treatment and / or prevention of a disease in a subject, wherein GLP-l-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein PAM and / or GLP-l-Gly may be in modified or unmodified form.

[0523] 2. GLP-l-Gly for use in treatment and / or prevention of a disease in a subject according to embodiment 1, wherein GLP-l-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM), wherein said subject is in need of elevated level of GLP-1- NH2 (GLP-1 -amide) in the circulation wherein elevated level means by at least 10 pg / mL, preferably by at least 20 pg / mL, more preferably by at least 30 pg / mL, more preferably by at least 40 pg / mL, most preferably by at least 50 pg / mL.

[0524] 3. GLP-l-Gly for use in treatment and / or prevention of a disease in a subject according to embodiments 1 or 2, wherein GLP-l-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM), wherein said disease is characterized by a level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or an GLP-1 -Gly / GLP-1 -amide ratio above a threshold in a sample of bodily fluid of said subject and / or a GLP-1 -amide level below a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or a GLP-1- amide level below a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof above a threshold and / or a GLP-1 -amide level above a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or a GLP-1 -amide level above a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof above a threshold. GLP-1 -Gly for use in treatment and / or prevention of a disease in a subject according to any of embodiments 1 to 3, wherein GLP-1 -Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM), and wherein said disease is selected from the group comprising pulmonary hypertension and hypertension, type-2 diabetes, inflammatory bowel disease and / or Crohn's disease, sepsis and / or septic shock, cerebral infarction, mild cognitive impairment (MCI) and Alzheimer’s Disease (AD). GLP-1 -Gly for use in treatment and / or prevention of a disease in a subject according to any of embodiments 1 to 4, wherein GLP-1 -Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM) and in addition is to be used with Vitamin C. GLP-1 -Gly for use in treatment and / or prevention of a disease in a subject according to any of embodiments 1 to 5, wherein GLP-1 -Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM) and wherein GLP-1 -Gly and PAM are to be administered consecutively, or GLP-1 -Gly and PAM are to be administered simultaneously. Peptide-Gly for use in treatment and / or prevention of a disease in a subject according to any of the preceding embodiments, wherein Peptide-Gly is GLP-1 -Gly (SEQ ID No.: 39-40), wherein GLP-1 -Gly (SEQ ID No.: 39-40) is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), and wherein said disease is type-2 diabetes. GLP-1 -Gly for use in treatment and / or prevention of a disease in a subject according to any of embodiments 1 to 7, wherein GLP-1 -Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM) and wherein the ratio of GLP-l-Gly and the GLP-1 -amide in a bodily fluid is elevated over 1.5, preferably elevated over 1.6, more preferably elevated over 1.7, more preferably is elevated over 1.8, more preferable is elevated over 1.9, more preferably is higher than 2 and / or the concentration of PAM (SEQ ID No. 1-10) in a bodily fluid is equal or below 100 ng / mL (100 ng / mL to 0 ng / mL), preferably equal or below 90 ng / mL (90 ng / mL to 0 ng / mL), more preferably equal or below 80 ng / mL (80 ng / mL to 0 ng / mL), more preferably equal or below 60 ng / mL (60 ng / mL to 0 ng / mL), more preferably equal or below 50 ng / mL (50 ng / mL to 0 ng / mL) and most preferably equal or below 40 ng / mL (40 ng / mL to 0 ng / mL) and / or the activity of PAM (SEQ ID No. 1-10) in a bodily fluid is equal or below 24 Units / L, more preferably equal or below 20 Units / L, more preferably equal or below 16 Units / L, more preferably equal or below 14 Units / L and most preferably equal or below 10 Units / L and / or

[0525] GLP-1 -amide level is equal or below 20 pg / mL, preferably equal or below 15 pg / mL, preferably equal or below 10 pg / mL, preferably equal or below 5 pg / mL and / or

[0526] GLP-1 is equal or above 15 pg / mL, preferably equal or above 20 pg / mL, preferably equal or above 30 pg / mL, preferably is equal or above 50 pg / mL, and / or

[0527] GLP-1 -amide level is equal or above 70 pg / mL, irrespective of the status of PAM and / or GLP- l-Gly, and / or the concentration of PAM (SEQ ID No.: 1-10) is equal or above 80 ng / mL, preferably equal or above 100 ng / mL, irrespective of the GLP-1 -amide and GLP-l-Gly status, and / or the activity of PAM (SEQ ID No.: 1-10) is equal or above 14 Units / L, preferably equal or above 20 units / L, preferably equal or above 24 units / L, irrespective of a GLP-1 -amide and GLP-l-Gly level. GLP-l-Gly for use in treatment and / or prevention of a disease in a subject according to any of embodiments 1 to 8, wherein GLP-l-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM) and wherein GLP-l-Gly is applied at a dosage of 0.1-500 pg / kg, preferably 1-400 pg / kg, more preferably 2-300 pg / mL, more preferably 2-300 pg / kg, more preferably 3-200 pg / kg, more preferably 5-100 pg / kg and wherein PAM is applied at a dosage of 1.5 - 54 Units / kg, preferably 1.7-33 Units / kg, more preferably 1.8-26 Units / kg, more preferably 1.9-20.6 Units / kg, more preferably 2-14 Units / kg. GUP-l-Gly for use in treatment and / or prevention of a disease in a subject according to any of embodiments 1 to 9, wherein GLP-l-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM) and in addition is to be used with Vitamin C and wherein Vitamin C is applied at a dosage of 1-10000 mg / kg, preferably 2-8000 mg / kg, more preferably 3-6000 mg / kg, more preferably 4-4000 mg / kg, more preferably 5-2000 mg / kg, more preferably 10-1000 mg / kg. GUP-l-Gly for use in treatment and / or prevention of a disease in a subject according to any of embodiments 1 to 10, wherein GUP-l-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM) wherein said PAM is modified via amino acid manipulations, preferably via fusion to Albumins, e.g. serum Albumin or recombinant serum Albumin, more preferably via non covalent binding to serum Albumin due to a conjugated fatty acid chain to PAM, mor preferably via fusion with IgG Fc regions or Transferrin, more preferably modified to act as PEG-based carrier prodrug, most preferably via post-translational modifications attaching natural or synthetic polymers, whereas the natural or synthetic polymer to be used is HAP, preferably EEP, more preferably PAS, more preferably PSA, more preferably GLK, more preferably XTEN and most preferably PEG. GLP-l-Gly for use in treatment and / or prevention of a disease in a subject according to any of embodiments 1 to 11, wherein GLP-l-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM) wherein said PAM is modified by pegylation. Pharmaceutical combination or pharmaceutical composition comprising GLP-l-Gly and peptidylglycine a-amidating monooxygenase (PAM), wherein said combination may be a kits of part combination with at least two pharmaceutical compositions, wherein one of which comprises GLP-l-Gly and wherein the other comprises PAM or wherein said pharmaceutical composition comprises both GLP-l-Gly and peptidylglycine a-amidating monooxygenase (PAM), and wherein PAM and / or GLP-l-Gly may be in modified or unmodified form. Pharmaceutical combination or pharmaceutical composition according to embodiment 13, wherein said combination or composition comprises in addition Vitamin C. Pharmaceutical combination or pharmaceutical composition according to any of embodiment 13 or 14, wherein GLP-l-Gly is applied at a dosage of 0.1-500 pg / kg, preferably 1-400 pg / kg, more preferably 2-300 pg / mL, more preferably 2-300 pg / kg, more preferably 3-200 pg / kg, more preferably 5-100 pg / kg and wherein PAM is applied at a dosage of 1.5 - 54 Units / kg, preferably 1.7-33 Units / kg, more preferably 1.8-26 Units / kg, more preferably 1.9-20.6Units / kg, more preferably 2-14.0 Units / kg

[0528] 16. Pharmaceutical combination or pharmaceutical composition according to any of embodiments 13 or 15, wherein said combination or composition comprises in addition Vitamin C and wherein Vitamin C is applied at a dosage of 1-10000 mg / kg, preferably 2-8000 mg / kg, more preferably 3-6000 mg / kg, more preferably 4-4000 mg / kg, more preferably 5-2000 mg / kg, more preferably 10-1000 mg / kg.

[0529] 17. Pharmaceutical combination or pharmaceutical composition according to any of embodiment 13 or 16, wherein GLP-l-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) wherein said PAM is modified via amino acid manipulations, preferably via fusion to Albumins, e.g. serum Albumin or recombinant serum Albumin, more preferably via non covalent binding to serum Albumin due to a conjugated fatty acid chain to PAM, mor preferably via fusion with IgG Fc regions or Transferrin, preferably modified to act as PEG-based carrier prodrug, most preferably via post-translational modifications attaching natural or synthetic polymers, whereas the natural or synthetic polymer to be used is HAP, preferably EEP, more preferably PAS, more preferably PSA, more preferably GLK, more preferably XTEN and most preferably PEG.

[0530] The following embodiments therefore further form part of the present invention:

[0531] 1. A method of treating and / or preventing a disease in a subject, said method comprising administering to said subject an effective amount of GLP-l-Gly, wherein GLP-l-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein PAM and / or GLP-l-Gly may be in modified or unmodified form.

[0532] 2. A method of treating and / or preventing a disease in a subject, said method comprising administering to said subject an effective amount of GLP-l-Gly according to embodiment 1, wherein GLP-l-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), wherein said subject is in need of elevated level of GLP-I-NH2 (GLP- 1 -amide) in the circulation wherein elevated level means by at least 10 pg / mL, preferably by at least 20 pg / mL, more preferably by at least 30 pg / mL, more preferably by at least 40 pg / mL, most preferably by at least 50 pg / mL. A method of treating and / or preventing a disease in a subject, said method comprising administering to said subject an effective amount of GLP-l-Gly according to embodiment 1 or 2, wherein GLP-l-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), wherein said disease is characterized by a level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or a GLP-l-Gly / corresponding GLP-l-amide ratio above a threshold in a sample of bodily fluid of said subject and / or a GLP-l-amide level below a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or a GLP-l-amide level below a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof above a threshold and / or a GLP-l-amide level above a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or a GLP-l-amide level above a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof above a threshold. A method of treating and / or preventing a disease in a subject, said method comprising administering to said subject an effective amount of GLP-l-Gly according to any of embodiments 1 to 3, wherein GLP-l-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM), and wherein said disease is selected from the group comprising pulmonary hypertension and hypertension, type-2 diabetes, inflammatory bowel disease and / or Crohn's disease, sepsis and / or septic shock, cerebral infarction, mild cognitive impairment (MCI) and Alzheimer’s Disease (AD). A method of treating and / or preventing a disease in a subject, said method comprising administering to said subject an effective amount of GLP-l-Gly according to any of embodiments 1 to 4, wherein GLP-l-Gly is GLP-l-Gly (SEQ ID No.: 39-40), wherein GLP-l- Gly (SEQ ID No.: 39-40) is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), and wherein said disease is type-2 diabetes. A method of treating and / or preventing a disease in a subject, said method comprising administering to said subject an effective amount of GLP-l-Gly according to any of embodiments 1 to 5, wherein GLP-l-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM) and in addition is to be used with Vitamin C. A method of treating and / or preventing a disease in a subject, said method comprising administering to said subject an effective amount of GLP-l-Gly according to any of embodiments 1 to 6, wherein GLP-l-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM) and wherein GLP-l-Gly and PAM are to be administered consecutively, or GLP-l-Gly and PAM are to be administered simultaneously. A method of treating and / or preventing a disease in a subject, said method comprising administering to said subject an effective amount of GLP-l-Gly according to any of embodiments 1 to 7, wherein the ratio of GLP-l-Gly and the corresponding GLP-1 -amide in a bodily fluid is elevated over 1.5, preferably elevated over 1.6, more preferably elevated over 1.7, more preferably is elevated over 1.8, more preferable is elevated over 1.9, more preferably is higher than 2 and / or the concentration of PAM (SEQ ID No. 1-10) in a bodily fluid is equal or below 100 ng / mL (100 ng / mL to 0 ng / mL), preferably equal or below 90 ng / mL (90 ng / mL to 0 ng / mL), more preferably equal or below 80 ng / mL (80 ng / mL to 0 ng / mL), more preferably equal or below 60 ng / mL (60 ng / mL to 0 ng / mL), more preferably equal or below 50 ng / mL (50 ng / mL to 0 ng / mL) and most preferably equal or below 40 ng / mL (40 ng / mL to 0 ng / mL) and / or the activity of PAM (SEQ ID No. 1-10) in a bodily fluid is equal or below 24 Units / L, more preferably equal or below 20 Units / L, more preferably equal or below 16 Units / L, more preferably equal or below 14 Units / L, and most preferably equal or below 10 Units / L and / or

[0533] GLP-1 -amide level is equal or below 20 pg / mL, preferably equal or below 15 pg / mL, preferably equal or below 10 pg / mL, preferably equal or below 5 pg / mL and / or

[0534] GLP-l-Gly is equal or above 15 pg / mL, preferably equal or above 20 pg / mL, preferably equal or above 30 pg / mL, preferably is equal or above 50 pg / mL, and / or

[0535] GLP-1 -amide level is equal or above 70 pg / mL, irrespective of the status of PAM and / or GLP- l-Gly, and / or the concentration of PAM (SEQ ID No.: 1-10) is equal or above 80 ng / mL, preferably equal or above 100 ng / mL, irrespective of the corresponding GLP-1 -amide and GLP-l-Gly status, and / or the activity of PAM (SEQ ID No.: 1-10) is equal or above 14 Units / L, preferably equal or above 20 units / L, preferably equal or above 24 units / L, irrespective of a corresponding GLP-1 -amide and GLP-1 -Gly level. A method of treating and / or preventing a disease in a subject, said method comprising administering to said subject an effective amount of GLP-1 -Gly according to any of embodiments 1 to 8, wherein GLP-l-Gly is applied at a dosage of 0.1-500 pg / kg, preferably 1- 400 pg / kg, more preferably 2-300 pg / mL, more preferably 2-300 pg / kg, more preferably 3-200 pg / kg, more preferably 5-100 pg / kg and wherein PAM is applied at a dosage of 1.5 - 54 Units / kg, preferably 1.7-33 Units / kg, more preferably 1.8-26 Units / kg, more preferably 1.9- 20.6Units / kg, more preferably 2-14.0 Units / kg. A method of treating and / or preventing a disease in a subject, said method comprising administering to said subject an effective amount of GLP-l-Gly according to any of embodiments 1 to 9, wherein GLP-l-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM) and in addition is to be used with Vitamin C and wherein Vitamin C is applied at a dosage of 1-10000 mg / kg, preferably 2-8000 mg / kg, more preferably 3-6000 mg / kg, more preferably 4-4000 mg / kg, more preferably 5-2000 mg / kg, more preferably 10-1000 mg / kg. A method of treating and / or preventing a disease in a subject, said method comprising administering to said subject an effective amount of GLP-l-Gly according to any of embodiments 1 to 10, wherein GLP-l-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM) wherein said PAM is modified via amino acid manipulations, preferably via fusion to Albumins, e.g. serum Albumin or recombinant serum Albumin, more preferably via non covalent binding to serum Albumin due to a conjugated fatty acid chain to PAM, mor preferably via fusion with IgG Pc regions or Transferrin, preferably modified to act as PEG-based carrier prodrug, most preferably via post-translational modifications attaching natural or synthetic polymers, whereas the natural or synthetic polymer to be used is HAP, preferably ELP, more preferably PAS, more preferably PSA, more preferably GLK, more preferably XTEN and most preferably PEG. A method of treating and / or preventing a disease in a subject, said method comprising administering to said subject an effective amount of GLP-l-Gly according to any of embodiments 1 to 11, wherein GLP- 1-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM) wherein said PAM is modified by pegylation.

[0536] The following embodiments also form part of the present invention:

[0537] 1. Peptide-Gly for use in treatment and / or prevention of a disease in a subject, wherein Peptide- Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein PAM and / or Peptide-Gly may be in modified or unmodified form.

[0538] 2. Peptide-Gly for use in treatment and / or prevention of a disease in a subject according to embodiment 1, wherein Peptide-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM) and wherein Peptide-Gly is selected from the group comprising

[0539] • from the group of Adrenomedullin family of peptides: Adrenomedullin-2-Gly; Intermedin-Short-Gly and Proadrenomedullin N-20 terminal peptide-Gly,

[0540] • from the Bombesin / neurimedin-B / Ranatensin family of peptides: Gastrin-releasing peptide-Gly; Neuromedin-C-Gly; Neuromedin-B-Gly and Nouromedin-B-32-Gly,

[0541] • from the Calcitonin family of peptides: Calcitonin-Gly; Calcitonin gene-related peptide (CGRP) 1-Gly; CGRP 2-Gly and Islet amyloid polypeptide-Gly,

[0542] • from the Chromogranin / Secretogranin family of peptides: Chromogranin-A-Gly; ER- 37-Gly; GR-44-Gly and Pancreastatin-Gly,

[0543] • from the FMRF-amide related peptide family of peptides: Neuropeptide AF-Gly; Neuropeptide FF-Gly; Neuropeptide SF-Gly Neuropeptide NPSF-Gly; Neuropeptide RFRP-1- Gly; Prolactin-releasing peptide PrRP20-Gly and Prolactin-releasing peptide PrRP31-Gly,

[0544] • from the Gastrin / Cholecystokinin family of peptides: Cholecystokinin -Gly; Cholecystokinin -12-Gly; Cholecystokinin -18-Gly; Cholecystokinin -25-Gly; Cholecystokinin -33-Gly; Cholecystokinin -39-Gly; Cholecystokinin -5-Gly; Cholecystokinin -58-Gly; Cholecystokinin -8-Gly; Cholecystokinin -7-Gly; Big-gastrin-Gly; Gastrin-Gly; Gastrin-14- Gly; Gastrin-52-Gly; Gastrin-6-Gly and Gastrin-71 -Gly,

[0545] • from the Glucagon family of peptides: Glucagon-like peptide (GLP) 1-Gly; GLP-l-Gly (7-37); Pituitary adenylate cyclase-activating polypeptide (PACP) 27-Gly; PACAP 38-Gly; Secretin-Gly; Somatoliberin-Gly; Intestinal Peptide PHM-27-Gly; Intestinal Peptide PHV-42- Gly and Vasoactive intestinal Peptide (VIP)-Gly,

[0546] • from the gonadotropin-releasing hormone family of peptides: Ginadoliberin- 1-Gly; Gonadoliberin-2-Gly; Progonadoliberin- 1-Gly and Progonadoliberin-2-Gly, • from the Kisspeptin family of peptides: Kisspeptin-10-Gly; Kisspeptin-13-Gly; Kisspeptin-14-Gly; Metastin-Gly and Metastasis-suppressor KISS- 1-Gly,

[0547] • from the Melanin-concentrating hormone family of peptides: Neuropeptide-glutamic acid-isoleucine-Gly and Pro-MCH-Gly,

[0548] • from the Neuromedin family of peptides: Neuromedin-S-Gly and Neuromedin-U-25- Gly,

[0549] • from the Neuropeptide-Y (NPY) family of peptides: Neuropeptide Y-Gly; Pancreatic hormone-Gly; Peptide YY-Gly and Peptide YY( 3-36)-Gly,

[0550] • from the opioid family of peptides: Deltrophin I-Gly,

[0551] • from the Orexin family of peptides: Orexin-A-Gly and Orexin-B-Gly,

[0552] • from the Proopiomelanocortin (POMC) family of peptides: Corticotropin-Gly; alpha- Melanotropin-Gly; gamma-Melanotropin-Gly and N-Terminal peptide of proopiomelanocortin- Gly (NPP-Gly),

[0553] • from the RFamide neuropeptide family of peptides: Orexigenic neuropeptide QRF-Gly (QRF-Gly),

[0554] • from the Sauvagine / corticotropin-releasing factor / urotensin I family of peptides: Urocortin-Gly and Urocortin-3-Gly,

[0555] • from the Tachykinin family of peptides: Substance P-Gly (RPKPQQFFGLMG) and Substance P-Gly (RPKPQFFGLMG); Neurokinin A-Gly; Neuropeptide K-Gly and Neurokinin- B-Gly,

[0556] • from the Thyrotropin Releasing Hormone (TRH) family of peptides: pro-TRH-Gly and TRH-Gly,

[0557] • from the Vasopressin / Oxytocin family of peptides: Oxytocin-Gly and Arg-Vasopressin- Gly,

[0558] • from the VGF nerve growth factor inducible protein (VGF) family of peptides: Antimicrobial peptide VGF-Gly (aa554-578) and Neurosecretory protein VGF-Gly. Peptide-Gly for use in treatment and / or prevention of a disease in a subject according to embodiments 1 or 2, wherein Peptide-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM) and wherein Peptide-Gly is Adrenomedullin (ADM)-Gly and wherein ADM-Gly may be in modified or unmodified form. Peptide-Gly for use in treatment and / or prevention of a disease in a subject according to embodiments 1 to 3, wherein Peptide-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM), wherein said subject is in need of elevated level of Peptide- NH2 (Peptide-amide) in the circulation wherein elevated level means by at least 10 pg / mL, preferably by at least 20 pg / mL, more preferably by at least 30 pg / mL, more preferably by at least 40 pg / mL, most preferably by at least 50 pg / mL. Peptide-Gly for use in treatment and / or prevention of a disease in a subject according to embodiment 1 to 4, wherein Peptide-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM), wherein said disease is characterized by a level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or a Peptide-Gly / corresponding Peptide-amide ratio above a threshold in a sample of bodily fluid of said subject and / or a Peptide- amide level below a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or a Peptide- amide level below a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof above a threshold and / or a Peptide- amide level above a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or a Peptide- amide level above a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof above a threshold. Peptide-Gly for use in treatment and / or prevention of a disease in a subject according to any of embodiments 1 to 5, wherein Peptide-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM), and wherein said disease is associated with endothelial barrier dysfunction and / or blood-brain barrier dysfunction. Peptide-Gly for use in treatment and / or prevention of a disease in a subject according to any of embodiments 1 to 6, wherein Peptide-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM), and wherein said disease is selected from the group comprising pulmonary hypertension and hypertension, type-2 diabetes, inflammatory bowel disease and / or Crohn's disease, sepsis and / or septic shock, cerebral infarction, mild cognitive impairment (MCI), dementia including cerebrovascular dementia, dementia with Lewy bodies (DLB), frontotemporal dementia (FTD) and Alzheimer’s Disease (AD). Peptide-Gly for use in treatment and / or prevention of a disease in a subject according to any of the preceding embodiments, wherein Peptide-Gly is GLP-l-Gly (SEQ ID No.: 39-40), wherein GLP-l-Gly (SEQ ID No.: 39-40) is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), and wherein said disease is type-2 diabetes. Peptide-Gly for use in treatment and / or prevention of a disease in a subject according to any of embodiments 1 to 8, wherein Peptide-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM) and in addition is to be used with Vitamin C. Peptide-Gly for use in treatment and / or prevention of a disease in a subject according to any of embodiments 1 to 9, wherein Peptide-Gly is to be used in combination with peptidylglycine a- amidating monooxygenase (PAM) and wherein Peptide-Gly and PAM are to be administered consecutively, or Peptide-Gly and PAM are to be administered...

Claims

Claims1. Peptide-Gly for use in treatment and / or prevention of a disease in a subject, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein PAM and / or Peptide-Gly may be in modified or unmodified form.

2. Peptide-Gly for use in treatment and / or prevention of a disease in a subject according to claim 1, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein Peptide-Gly is selected from the group comprising• from the group of Adrenomedullin family of peptides: Adrenomedullin-2-Gly; Intermedin- Short-Gly and Proadrenomedullin N-20 terminal peptide-Gly,• from the Bombesin / neurimedin-B / Ranatensin family of peptides: Gastrin-releasing peptide- Gly; Neuromedin-C-Gly; Neuromedin-B-Gly and Nouromedin-B-32-Gly,• from the Calcitonin family of peptides: Calcitonin-Gly; Calcitonin gene-related peptide (CGRP) 1-Gly; CGRP 2-Gly and Islet amyloid polypeptide-Gly,• from the Chromogranin / Secretogranin family of peptides: Chromogranin-A-Gly; ER-37-Gly; GR-44-Gly and Pancreastatin-Gly,• from the FMRF-amide related peptide family of peptides: Neuropeptide AF-Gly; Neuropeptide FF-Gly; Neuropeptide SF-Gly Neuropeptide NPSF-Gly; Neuropeptide RFRP-l-Gly; Prolactinreleasing peptide PrRP20-Gly and Prolactin-releasing peptide PrRP31-Gly,• from the Gastrin / Cholecystokinin family of peptides: Cholecystokinin -Gly; Cholecystokinin - 12-Gly; Cholecystokinin -18-Gly; Cholecystokinin -25-Gly; Cholecystokinin -33-Gly; Cholecystokinin -39-Gly; Cholecystokinin -5-Gly; Cholecystokinin -58-Gly; Cholecystokinin - 8-Gly; Cholecystokinin -7-Gly; Big-gastrin-Gly; Gastrin-Gly; Gastrin- 14-Gly; Gastrin-52-Gly; Gastrin-6-Gly and Gastrin-71 -Gly,• from the Glucagon family of peptides: Glucagon-like peptide (GLP) 1-Gly; GLP-l-Gly (7-37); Pituitary adenylate cyclase-activating polypeptide (PACP) 27-Gly; PACAP 38-Gly; Secretin- Gly; Somatoliberin-Gly; Intestinal Peptide PHM-27-Gly; Intestinal Peptide PHV-42-Gly and Vasoactive intestinal Peptide (VIP)-Gly,• from the gonadotropin-releasing hormone family of peptides: Ginadoliberin- 1-Gly; Gonadoliberin-2-Gly; Progonadoliberin- 1-Gly and Progonadoliberin-2-Gly,• from the Kisspeptin family of peptides: Kisspeptin-10-Gly; Kisspeptin-13-Gly; Kisspeptin- 14- Gly; Metastin-Gly and Metastasis-suppressor KISS- 1-Gly,• from the Melanin-concentrating hormone family of peptides: Neuropeptide-glutamic acid- isoleucine-Gly and Pro-MCH-Gly,• from the Neuromedin family of peptides: Neuromedin-S-Gly and Neuromedin-U-25-Gly,• from the Neuropeptide-Y (NPY) family of peptides: Neuropeptide Y-Gly; Pancreatic hormone- Gly; Peptide YY-Gly and Peptide YY( 3-36)-Gly,• from the opioid family of peptides: Deltrophin I-Gly,• from the Orexin family of peptides: Orexin-A-Gly and Orexin-B-Gly,• from the Proopiomelanocortin (POMC) family of peptides: Corticotropin-Gly; alpha- Melanotropin-Gly; gamma-Melanotropin-Gly and N-Terminal peptide of proopiomelanocortin- Gly (NPP-Gly),• from the RFamide neuropeptide family of peptides: Orexigenic neuropeptide QRF-Gly (QRF- Gly),• from the Sauvagine / corticotropin-releasing factor / urotensin I family of peptides: Urocortin-Gly and Urocortin-3-Gly,• from the Tachykinin family of peptides: Substance P-Gly (RPKPQQFFGLMG) and Substance P-Gly (RPKPQFFGLMG); Neurokinin A-Gly; Neuropeptide K-Gly and Neurokinin-B-Gly,• from the Thyrotropin Releasing Hormone (TRH) family of peptides: pro-TRH-Gly and TRH- Gly,• from the Vasopressin / Oxytocin family of peptides: Oxytocin-Gly and Arg-Vasopressin-Gly,• from the VGF nerve growth factor inducible protein (VGF) family of peptides: Antimicrobial peptide VGF-Gly (aa554-578) and Neurosecretory protein VGF-Gly.

3. Peptide-Gly for use in treatment and / or prevention of a disease in a subject according to claims 1 or2, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein Peptide-Gly is Adrenomedullin (ADM)-Gly and wherein ADM-Gly may be in modified or unmodified form.

4. Peptide-Gly for use in treatment and / or prevention of a disease in a subject according to claims 1 to3, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), wherein said subject is in need of elevated level of Peptide-bTT (Peptideamide) in the circulation wherein elevated level means by at least 10 pg / mL, preferably by at least 20 pg / mL, more preferably by at least 30 pg / mL, more preferably by at least 40 pg / mL, most preferably by at least 50 pg / mL.

5. Peptide-Gly for use in treatment and / or prevention of a disease in a subject according to claim 1 to4, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), wherein said disease is characterized by a level of PAM and / or its isoforms and / or fragments thereof below a threshold and / ora Peptide-Gly / corresponding Peptide-amide ratio above a threshold in a sample of bodily fluid of said subject and / or a Peptide- amide level below a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or a Peptide- amide level below a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof above a threshold and / or a Peptide- amide level above a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof below a threshold and / or a Peptide- amide level above a certain threshold in combination with level of PAM and / or its isoforms and / or fragments thereof above a threshold.

6. Peptide-Gly for use in treatment and / or prevention of a disease in a subject according to any of claims 1 to 5, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), and wherein said disease is associated with endothelial barrier dysfunction and / or blood-brain barrier dysfunction.

7. Peptide-Gly for use in treatment and / or prevention of a disease in a subject according to any of claims 1 to 6, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), and wherein said disease is selected from tire group comprising pulmonary hypertension and hypertension, type-2 diabetes, inflammatory bowel disease and / or Crohn's disease, sepsis and / or septic shock, cerebral infarction, mild cognitive impairment (MCI), dementia including cerebrovascular dementia, dementia with Lewy bodies (DLB), frontotemporal dementia (FTD) and Alzheimer’s Disease (AD).

8. Peptide-Gly for use in treatment and / or prevention of a disease in a subject according to any of the preceding claims, wherein Peptide-Gly is GLP-l-Gly (SEQ ID No.: 39-40), wherein GLP-l-Gly (SEQ ID No.: 39-40) is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM), and wherein said disease is type-2 diabetes.

9. Peptide-Gly for use in treatment and / or prevention of a disease in a subject according to any of claims 1 to 8, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and in addition is to be used with Vitamin C.

10. Peptide-Gly for use in treatment and / or prevention of a disease in a subject according to any of claims 1 to 9, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein Peptide-Gly and PAM are to be administered consecutively, or Peptide-Gly and PAM are to be administered simultaneously.

11. Peptide-Gly for use in treatment and / or prevention of a disease in a subject according to any of claims 1 to 10, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein the ratio of Peptide-Gly and the corresponding Peptide-amide in a bodily fluid is elevated over 1.5, preferably elevated over 1.6, more preferably elevated over 1.7, more preferably is elevated over 1.8, more preferable is elevated over 1.9, more preferably is higher than 2 and / or the concentration of PAM (SEQ ID No. 1-10) in a bodily fluid is equal or below 100 ng / mL preferably equal or below 90 ng / mL, more preferably equal or below 80 ng / mL, more preferably equal or below 60 ng / mL, more preferably equal or below 50 ng / mL and most preferably equal or below 40 ng / mL and / or the activity of PAM (SEQ ID No. 1-10 ) in a bodily fluid is equal or below 24 Units / L, more preferably equal or below 20 Units / L, more preferably equal or below 16 Units / L, more preferably equal or below 14 Units / L, and most preferably equal or below 10 Units / L and / orPeptide-amide level is equal or below 20 pg / mL, preferably equal or below 15 pg / mL, preferably equal or below 10 pg / mL, preferably equal or below 5 pg / mL and / orPeptide-Gly is equal or above 15 pg / mL, preferably equal or above 20 pg / mL, preferably equal or above 30 pg / mL, preferably is equal or above 50 pg / mL, and / orPeptide-amide level is equal or above 70 pg / mL, irrespective of the status of PAM and / or Peptide-Gly, and / or the concentration of PAM (SEQ ID No.: 1-10) is equal or above 80 ng / mL, preferably equal or above 100 ng / mL, irrespective of the corresponding Peptide-amide and Peptide-Gly status, and / or the activity of PAM (SEQ ID No.: 1-10) is equal or above 14 Units / L, preferably equal or above 20 units / L, preferably equal or above 24 units / L, irrespective of a corresponding Peptide-amide and Peptide-Gly level.

12. Peptide-Gly for use in treatment and / or prevention of a disease in a subject according to any of claims 1 to 11, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and wherein Peptide-Gly is applied at a dosage of 0.1-500 pg / kg, preferably 1-400 pg / kg, more preferably 2-300 pg / mL, more preferably 2-300 pg / kg, more preferably 3-200 pg / kg, more preferably 5-100 pg / kg and wherein PAM is applied at a dosage of 1.5 - 54 Units / kg, preferably 1.7-33 Units / kg, more preferably 1.8-26 Units / kg, more preferably 1.9-20.6 Units / kg, more preferably 2-14 Units / kg13. Peptide-Gly for use in treatment and / or prevention of a disease in a subject according to any of claims 1 to 12, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) and in addition is to be used with Vitamin C and wherein Vitamin C is applied at a dosage of 1-10000 mg / kg, preferably 2-8000 mg / kg, more preferably 3-6000 mg / kg, more preferably 4-4000 mg / kg, more preferably 5-2000 mg / kg, more preferably 10-1000 mg / kg.

14. Peptide-Gly for use in treatment and / or prevention of a disease in a subject according to any of claims 1 to 13, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) wherein said PAM is modified via amino acid manipulations, preferably via fusion to Albumins, e.g. serum Albumin or recombinant serum Albumin, more preferably via non covalent binding to serum Albumin due to a conjugated fatty acid chain to PAM, mor preferably via fusion with IgG Fc regions or Transferrin, more preferably modified to act as PEG-based carrier prodrug, most preferably via post-translational modifications attaching natural or synthetic polymers, whereas the natural or synthetic polymer to be used is HAP, preferably EEP, more preferably PAS, more preferably PSA, more preferably GLK, more preferably XTEN and most preferably PEG.

15. Peptide-Gly for use in treatment and / or prevention of a disease in a subject according to any of claims 1 to 14, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) wherein said PAM is modified by pegylation.

16. Pharmaceutical combination or pharmaceutical composition comprising Peptide-Gly and peptidylglycine a-amidating monooxygenase (PAM), wherein said combination may be a kits of part combination with at least two pharmaceutical compositions, wherein one of which comprises Peptide-Gly and wherein the other comprises PAM or wherein said pharmaceutical composition comprises both Peptide-Gly and peptidylglycine a-amidating monooxygenase (PAM), and wherein PAM and / or Peptide-Gly may be in modified or unmodified form.

17. Pharmaceutical combination or pharmaceutical composition according to claim 16, wherein said combination or composition comprises in addition Vitamin C.

18. Pharmaceutical combination or pharmaceutical composition according to any of claim 16 or 17, wherein Peptide-Gly is applied at a dosage of 0.1-500 pg / kg, preferably 1-400 pg / kg, more preferably 2-300 pg / mL, more preferably 2-300 pg / kg, more preferably 3-200 pg / kg, more preferably 5-100 pg / kg and wherein PAM is applied at a dosage of 1.5 - 54 Units / kg, preferably 1.7-33 Units / kg, more preferably 1.8-26 Units / kg, more preferably 1.9-20.6 Units / kg, more preferably 2-14.0 Units / kg.

19. Pharmaceutical combination or pharmaceutical composition according to any of claims 16 to 18, wherein said combination or composition comprises in addition Vitamin C and wherein Vitamin C is applied at a dosage of 1-10000 mg / kg, preferably 2-8000 mg / kg, more preferably 3-6000 mg / kg, more preferably 4-4000 mg / kg, more preferably 5-2000 mg / kg, more preferably 10-1000 mg / kg.

20. Pharmaceutical combination or pharmaceutical composition according to any of claim 16 to 19, wherein Peptide-Gly is to be used in combination with peptidylglycine a-amidating monooxygenase (PAM) wherein said PAM is modified via amino acid manipulations, preferably via fusion to Albumins, e.g. serum Albumin or recombinant serum Albumin, more preferably via non covalent binding to serum Albumin due to a conjugated fatty acid chain to PAM, mor preferably via fusion with IgG Fc regions or Transferrin, preferably modified to act as PEG-based carrier prodrug, most preferably via post-translational modifications attaching natural or synthetic polymers, whereas the natural or synthetic polymer to be used is HAP, preferably EEP, more preferably PAS, more preferably PSA, more preferably GLK, more preferably XTEN and most preferably PEG.

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