Indian Patents. 258244:"PEPTIDIC VASOPRESSIN RECEPTOR AGONISTS"

Title of Invention	"PEPTIDIC VASOPRESSIN RECEPTOR AGONISTS"
Abstract	The present invention relates to novel compounds, pharmaceutical compositions comprising the same, use of said compounds for the manufacture of a medicament for treatment of inter alia shock conditions as well as to a method for treatment of said conditions, wherein said compounds are administered The compounds are represented by the general formula (I), as further defined in the specification.

Full Text	PEPTIDIC VASOPRESSIN RECEPTOR AGONISTS Field of the Invention The present invention relates to novel compounds, pharmaceutical compositions comprising the same, use of said compounds for the manufacture of a medicament for treatment of inter alia shock conditions as well as to a method for treatment of said conditions, wherein said compounds Ere administered. Background Peptidic vasopressin Via receptor agonists, such as terlipressir., have recently (see e.g. O'Brian et al. , Lancet 359 ; £313):1209-10, June 4th, 2002) received increased attention for clinical use in treatment of critical care diseases and conditions, including shock-of hypovolemic (e.g. hemorrhagic) or vasodilatory (e.g. septic) origin, bleeding esophageal varices (BEV), hepa-torenal syndrome (HRS), cardiopulmonary resuscitation and anesthesia-induced hypotension. They have also beer, shown to have clinical use in the treatment of orthostatic hypotension, paracentesis-induced circulatory dysfunction, intra-opexative blood loss and blood loss associated with burn debridement and epistaxis, and for treatment of various ocular diseases by increasing lacrimation/tear formation. In treating critical care conditions it is highly desirable to control the arterial blood pressure, and the drug used is typically administered intravenously.. Continuous intravenous drug infusion at increasing or decreasing rates is a practical means of providing the desired degree of control. The attainment of so-called "steady state" plasma concentrations of drug depends on the elimination half life of the drug infused. It is generally recognised that steady state plasma concentration is achieved after a period of time equivalent to three times the elimination half life of the drug. To be prac- tical in a clinical setting the desired arterial blooci pressure at the steady state should be attained in about two hours, preferably in one hour or less. Via agonists with an elimination half life longer than 1 hour are therefore usually not considered useful for critical care treatment, A disadvantage of terlipressin in many critical care situations is its long duration of action, which makes it difficult to titrate its effect as the disease state changes. The efficacy of terlipressin at the human Via (hVia) receptor also needs to be improved e.g. to allow lower dosages in general. Also the compound known as F180 {cf. example 3 in US patent No. 5,459,236) has an inconveniently long duration of action to be considered for the treatment of most critical care conditions. Non-specific receptor agonist activity is the main disadvantage of other existing compounds, e.g. [Phe2,Orn8jOT (cf. example If in US patent No. 3,352,843) and arginine-vasopressin (AVP). Activity at related receptors such as Vlb, V.2 and oxytocin (OT) receptors may potentially generate undesirable side effects and safety concerns. As an example, V2 receptor activation may induce antidiuresis (cf. desmopressrn), release of coagulation/thromboly sis factors, and induce vasodilation/hypo-tension with reflex tachycardia. The latter side effect may also be induced by OT receptor agonist activity. It is an objective of the present invention to provide compounds that are especially useful in the treatment of critical care conditions. Disclosure of the Invention The present invention relates to compounds represented by the general formula (I) (SEQ ID NO RaN(CH2)nR2 CH2 H-Cys^ A-xIle. Is,Asn-Cys-Prox ^X^Gly-NHo (I H T E T N T o H o H wherein: Ar is an aryl group selected from aromatic carbocyclic ring systems, five- or six-membereci heteroaronaatic ring systems and bicyclic heteroaromatic ring systems; m is selected from 1, 2 and 3; n is selected from 0, 1, 2, 3 and 4; p is selected from 2, 3 and 4; Ri, R2 and Rs are independently selected from H, OK, alkyl, 0-alkyI and OC(0)-alkyl; alkyl is selected from Ci_6 straight and C^-E branched chain alkyl and optionally has at least one hydroxyl substituent; and when n«0, HI and R2 optionally together form a nitrogen containing ring structure comprising from 2 to 5 carbon atoms; with the proviso that when Ar is phenyl (amino acid no, 2 is Phe), m«=2r n=0 and H!=R2=fl (amirio acid no.. 4 is Gin) R3 is riot H when p is 3 or 4; and solvates and pharmaceutically acceptable salts thereof. Amino acid no. B is Ozn when R3=H and p=3, and Lys when R3=H and p=4. For the purposes of the present invention, the following terminology is used-Aromatic carbocyclic ring systems includes phenyl and xiaphthyl. A five-membered heteroaromatic ring system is a monocyclic aromatic ring system having five ring atoms, wherein 1, 2 or 3 ring atoms are independently selected from N, 0 and S. Preferred such ring systems are selected form a group consisting of thienyl, furyl, pyrrolyl, imidazolyl, thiazolyl, oxazolyl, pyrazolyl, isothiazolyl, isoxazolyl and tetrazolyl. A six-membered heteroaromatic ring system is a mono-cyclic aromatic ring system having six ring atoms, wherein 1, 2 cr 3 ring atoms are independently selected from N, 0 and S. It is preferably selected from a group consisting of pyridyl, pyrazinyl, pyrimidinyl, triazir.yl and pyridazinyl. A bicyclic heteroaromatic ring system is a ring system having two five- or six-membered heteroaromatic rings, or a phenyl and a five- or six-membered heteroaxo-matic ring, cr a phenyl and a hererocyclyl ring, or a five- or six-membered heteroaromatic ring and a heterocy-clyl ring; connected by a ring fusion, said bicyclic heteroaromatic ring system comprising 8 to 12 ring atoms, wherein 1, 2 or 3 of the ring a~oms are independently selected from N, 0 and S, It is preferably selected from a group consisting of indole, guinoline, tetrahydroguino-line, isoquinoline, tetrahydroisoguinoline, 1,4-benzo-dioxan, coumarin, benzofuran, Ir2-benzisoxazole, ben-zothiophene, benzoxazole, benzthiazole, benzimidazole, benzxriazole, pyrolizidine and quinoliziditie, A heterocyclyl or heterocyclic moiety is a saturated or partially saturated ring system having 3 to 7 ring atoms, wherein 1, 2 or 3 ring atoms are independently selected from N, 0 and S, Heterocyclyl moieties are preferably selected from a group consisting of aziridine, oxirane, thiirane, azetidine, oxetane, thietane, pyr-rolidine., pyrroline, imidazolidine, pyrazolidine, di-oxolaner tetrahydrofuranylf piperidine, pipexazine, mor-pholine(/ tet:rahydropyranylf 1,4-dioxanyl, homopiperid-homopiperazinyl and hexamethylene oxide. It deserves mentioning that e.g. also isopropyl and 2-rt-butyl groups are encompassed by the expression Ci_6 straight chain alkyl, as said expression is not related to the binding site of the straight chain in question. Ci_€ denotes having from one to six carbon atoms, including any number therebetween, and this nomenclature is used analogously herein. Examples of phermaceurically acceptable salts comprise acid addition salts, e.g. a salt formed by reaction wirh hydrohalogen acids, such as hydrochloric acid, and mineral acids, such as sulphuric acid, phosphoric acid and nitric acid, as well as aliphatic, alicyclic, aromatic or heterocyclic suiphonic or carboxylic acids, sucft as formic acid, acetic acid, propionic acid, succinic acid, glycolic acid, lactic. acid, malic acid, tartaric acid, citric acid, ascorbic acid, maleic acid, hydroxymaleic acid, pyruvic acid, p-hydroxybenzcic acid, embonic acid, methanesulphonic acid, ethanesulphonic acid, hydroxyethanesulphonic acid, halobenzenesulphonic acid, toluenesulphonic acid and naphtalenesulphonic acid. Ar is preferably selected from phenyl, 2- or 3-thier.yl, 2- or 3-furyl, 2-, 3- or 4-pyridyl and 2-, 4- or 5-thiazolyl, It is particularly preferred that RI is H. In preferred embodiments p is 2 or 3.. It is preferred to select R2 from H, OH, CH3, CH2CH3, CH{CH3)2, CEiCH2OH)2, CH(OH)CH3 (both enantiomers) , OCH3 and OCH2CH2OH. Moreover, it is preferred to select R3 from H, methyl, ethyl, n-propyl, i-propyl and i-amyl. In the most preferred embodiment, said compound having the formula (I) is selected from a group consisting of (SEQ ID NOs: 1-7, respectively, in order of appear- H-Cys-Phe-Ile-Hgn-Asn-Cys-Pro-Orn(i-Pr)-Gly-NH2 y H-Cys-Phe-Ile-Asn { (CH2) 3OH) -Asn-Cys-Pro-Orn-Gly-NH;, (2) H-Cys-Phe-Ile-Asn-Asn~Cys-Pro-Dbu-Gly-NH2 H-Cys-Phe-Ile-Asn (CH2CH3) -Asn-Cys-Pro-Dbu-Gly-NH2 I _ I (4 ) H-Cys-Phe-Ile-Gln-Asn-Cys-Pro-Orn (i-Pr) -Gly-NH2 and H-Cys-Phe-Ile-Gln-Asn-Cys-Pro-Orn(CH2CH3) -Gly-NH2, I I :6> H-Cys-Phe-Iie-Asn(CH3}2-Asn-Cys-Pro-Orn-Gly-NH2 I J (7) The number in parenthesis denotes the compound as referered to in the following. Furthermore the present invention relates to a compound as set forth above for use as a pharmaceutical. Accordingly, the present invention also relates to a pharmaceutical composition comprising a compound as set forth above as active ingredient in association with a pharmaceutically acceptable adjuvant, diluent or carrier. The pharmaceutical composition may be adapted for oral, intravenous, topical, intraperitoneal, nasal, buccal, sublingual or subcutaneous administration cr for administration via the respiratory tract e.g. in the. form of an aerosol or an air-suspended fine powder. The composition may thus for instance be in the form of tablets, capsules., powders, microparticles, granules, syrups, suspensions, solutions, transdermal patches or suppositories. It .should be noted that the composition according to the present invention may optionally include two or more of the above outlined compounds. The present pharmaceutical composition may optionally comprise e.g. at least one further additive selected from a disintegrating agent, binder, lubricant, flavoring agent, preservative, colorant and any mixture thereof. Examples of such and other additives are found in "%Handbook of Pharmaceutical Excipients"; Ed. A.H. Kibbe, 3rd Ed., American Pharmaceutical Association, USA and Pharmaceutical Press UK, 2000. The present pharmaceutical composition is most preferably adapted for parenteral administration. It may comprise a sterile aqueous preparation of the compounds of the invention preferably isotonic with the blood of the recipient. This aqueous preparation may be formulated according 10 kn.own methods using suitable dispersing or wetting agents and suspending agents. The injectable aqueous formulation Remestyp® (terlipressin) is exemplary of a suitable pharmaceutical formulation. The preparation may also be a sterile injectable solution or suspension in a diluent or solvent, for example as a solution in 1,3-butane diol. Water, Ringer's solution, and isotonic sodium chloride solution are exemplary acceptable diluents.. Sterile, fixed oils may be employed as a solvent or suspending medium. Bland fixed oils, including synthetic mono or di-glycerides, and fatty acids, such as oleic acid, may also be used. In addition, the present invention relates tc use of a compound as outlined above for the manufacture of a medicament for treatment of shock of hypovolemic or vasodi-latory origin, BEV, HRS, cardiopulmonary resuscitation, anesthesia-induced hypotension, orthostatic hypotension, paracentesis-induced circulatory dysfunction, intra-operative blood loss or blood loss associated with burn debridement and epistaxis, and for treatment of various ocular diseases by increasing lacrimation/tear formation. In another embodiment the invention relates to a method for treatment of shock of hypovolemic or vasodila-tory origin., BEV, .HRS, cardiopulmonary resuscitation, an- esthesia-induced hypotension, orthostatic hypotension, paracentesis-induced circulatory dysfunction, intra-operative blood loss or blood loss associated with burn debridement and epistaxis, and of various ocular diseases by increasing lacrimation/tear formation, wherein said method comprises administering to an animal, including human, patient of a therapeutically effective amount of a compound as outlined above. The typical dosage of the compounds according to the present invention varies within a wide range and will depend on various factors such as the individual needs of each patient and the route of administration. The dosage administered by infusion is generally within the range of 0.01-200 ug/kg body weight per hour. A physician of ordinary skill in the art will be able to optimise the dosage to the situation at hand. The abbreviations used are: Abu 2-aminobutyric acid Boc tert-butoxycarbonyl BOP benzotriazol-1-yloxy "trisdimethylamino- phosphonium hexafluorophosphate Dbu 2,4-diaminobutyric acid DCC N,N'-dicyclohexylcarbodiimide DCHA dicyclohexylamine DCM dichloromethane DIAD diisopxopyl diazodicarboxylate DIG W/W-diisopropylcarbodiimide DIEA N, N-diisopropyl-JV-ethylamine DMF Nr jW-dimethylformamide Fm 9-fluorenylmethyl Fmoc S-fluorenylmethoxycarbonyl Hgn homoglutamine Hmp 2-hydroxy-3-mercaptopropionic acid HOBt 1-hydroxybenzotriazole HPLC high performance liquid chromatography i iso Mint 4-methoxytrityl Mob p-methoxybenzyl MS mass spectrometry Orn ornithine Ph phenyl Pr propyl PyBOP benzotriazol-1-yloxy trispyrrolidine- phosphonium hexafluorophosphate o-NBS-Cl2-nitrobenzenesulfonyl chloride OT oxytocin Rt retention time TFA trifluoroacetic acid TIS triisopropylsilane TMOF trimethylorthoformate TPP triphenylphosphine Trt trityl VT vasotocin, [lie3] vasopressin Unless otherwise specified L-amino acids were used, and conventional amino acid terminology is adhered to. Experimental (synthesis) Amino acid derivatives and resins were purchased from commercial providers (Novabiochem, Bachem Peptide International and PepTech Corporation). Fmoc-Hgn-OH was synthesised according to literature (Wisniewski, K., Kolodziejczyk, A..S, Org.. Prep. P-roced. Int.. 1997, 29, 338-341). Other chemicals and solvents were provided from Sigma-Aldrich, Fisher Scientific and VWR. The compounds herein were synthesised by standard methods in solid phase peptide chemistry utilising both Fmoc and Boc methodology.. Unless otherwise provided, all reactions were performed at room temperature. In addition to the references cited supra, the following standard reference literature provides further guidance on general experimental set up, as well as on the availability of required starting material and reagents;: Kates, S.A., Albericio, F., Eds., Solid Phase Synthesis. A Practical Guide, Marcel Dekker, New York, Basel, 2000; Stewart, J.M., Young, J.D. Solid Phase Synthesis, Pierce Chemical Company, 1984; Bisello, et al., J. Biol. Chem, 199B, 273, 22496-22505; and Merrifield, J, Am. Chem, Soc. 1963, 85, 2149-2154. Purity of the synthesized peptide may be determined by analytical reversed phase HPLC. Structural integrity of the peptides may be confirmed using amino acid analysis and electrospray mass spectrometry. The peptides synthesised by Fmoc methodology were cleaved with a TFA/TIS/H20 96/2/2 (v/v/v) solution, and cleavage in Boc methodology was accomplished with 90% HF/10% aniscle (v/v) solution. Disulfide bridge (ring) formation was achieved by oxidation of linear peptides dissolved in 10% TFA (aq) with iodine. Peptides were purified by preparative H?LC in triethylammonium phosphate buffers (aq}. The compounds were finally converted to acetate salts using conventional HPLC methodology. The fractions with a purity exceeding 97% were pooled and lyophilised. Synthesis of peptides with alkylated side chain in position no, 8: The peptides were assembled with Fmoc methodology. The diamrno acid residue in position no, 8 was introduced with an acid labile (i.e. removable with a solution containing 1-2% TFA) protecting group, such as methoxy-trityl (Mmt; see Barlos, K. et al. in Peptides 1992, Schneider, C.H., Eberle, A.N., Eds., ESCOM Science Publishers 3-V.r 1993, pp 263-284). Resin bound peptide was treated with a DCM/TIS/TFA 93/5/2 (v/v/v) solution for the Mmt group removal. Reductive alkylation with acetone/NaBH(OAc)s provided the W-isopropyl peptide. To avoid undesirable N,N-d±alkylation in reductive alkylation in the above procedure, which .may occur when straight chain alkyl aldehydes are used, an alternative was developed, wherein after the Mmt removal the amino group was first derivatised with 2-nitrobenzenesulfonyl chloride (o-NBS-Cl; see Fukuyama, T.; Jow, C.-K.; Cheung, N. Tetrahedron Lett. 1995, 36, 6373-6374). The resulting sulphonamide was then alkylated with an appropriate alcohol under conventional Mitsunobu reaction conditions, typically utilising TPP/DIAD in 1,2-dimethoxyethane (Mitsunobu, 0. Synthesis 1981, 1-2-8) . The o-NBS-Cl group was subsequently removed with 5% potassium thiophenolate ±r. DMF, after which the peptide was cleaved from the resin. Synthesis of peptides with Ar-alkylated side chair, in position no. 4: The peptides were assembled with Boc methodology. The residue in position no. 4 was introduced in the sequence as 3oc-Asp(OFm)-OH. After complete peptide assembly the side chain protection was removed with 30% piperidine in DMF. The resulting free carboxylic group was converted to the desired amide by coupling with an appropriate amine mediated by PyBOP or BOP/DIEA. The K-terminal Boc group was then removed, followed by EF cleavage, cyclisation and purification by HPLC, Table 1 lists the compounds prepared by the above procedure, RI is H for all compounds except no- 7, where RI is CHa- An asterisk "" marks the most preferred embodiments. (Table Remove) 1. Compounds prepared with the formula (I\| (Table Remove) The following detailed examples are provided tc further illustrate the synthesis: Compound 1; [Phe2, Hgn4,0rn (i-Pr)e] VT: The amino acid derivatives used were Boc-Cys(Trt) -OH, Fmoc-Phe-OH, Fmoc-Ile-OH, Fmoc-Hgn-OH, Fmoc-Asn(Trt)-OK, Fraoc-CysETrt)-OHf Fmoc-Pro-OH, Fmoc-Orn(Mmt)-OH and Fmoc-Gly-OH. Fmoc-Hgn-OH was synthesised as mentioned above. Analytical HPLC was performed on a Waters 600 Liquid Chromatograph using a Vydac C18, 5u 4.6 x 250 mm, column at a flow rate of 2 ml/min. Preparative HPLC was performed on a Waters 2000 Liquid Chromatograph using a Prepak 47 x 300 mm cartridge at a flow rate of IOC ml/min, Final compound analysis was performed on a 1100 Agilent Liquid Chromatograph using a Vydac C18r 5ja 2,1 x 250 mm, column at a flow rate of 0.3 ml/min. Mass spectra were recorded on a Finnigan MAT spectrometer. The fully protected peptide resin was synthesised on an Applied Biosystems 9050 Peptide Synthesiser starting from 2 g (0.5 mmol) of Tentagel-S-RAM resin (Peptides International). DIC/HOBt mediated single couplings with a 4-fold excess of amino acid derivatives were performed. The Fmoc group was removed with 20% piperidine in DMF. Upon completion of the automated synthesis, the resin was transferred into a manual synthesis vessel and was treated with DCM/TIS/TFA 93/5/2 (v/v/v) solution (30 ml) for 2 x 1.5 hours for removal of the Mmt group. The resin was thoroughly washed with DCM and was subsequently suspended in 15 ml of 1,2-dichloroetehane/TMOF 1:1 (v/v) . 0.2 ml of acetone was then added followed by 0,6 g :;f NaBH(OAc)3. The suspension was shaken overnight and \:he resin was washed with methanol, DMF and DCM and dried in vacua. The resin was then treated with 30 ml of the TFA/TIS/%0 96/2/2 (v/v/v) solution for 1.5 hours and filtered off. The filtrate was evaporated and the crude linear peptide was precipitated with diethyl ether. The precipitate was immediately dissolved in 500 ml of 10% TFA (aq), and the peptide was oxidised by adding 0.1 M T2 in methanol to the magnetically stirred solution unzil yellow color persisted. Excess of iodine was reduced with ascorbic acid. The reaction mixture was then cooled with crushed ice and pH was adjusted to about 5 by'adding concentrated ammonia (aq) . The mixture was loaded onto an HPLC column and purified using a triethylammonium phosphate buffer with pH 5.2. The compound was eluted with a gradient of acetonitrile. The fractions with a purity exceeding 97% were pooled, and the resulting solution was diluted with 2 volumes of water. The solution was reloaded onto the column which was then washed with 2 1 of 0,1 M ammonium acetate (aq) and equilibrated with 2% acetic acid (aq). The compound was eluted with a fast (3%/min) gradient of acetonitrile. The fractions containing the desired product were pooled and lyophillsed.. 168 mg (-30% yield) of white amorphous powder was obtained. HPLC: Rt=8.5 min, gradient; 20-. Compound 4? [Phe2,Asn (Et)4, Dbu8]VT : The amino acid derivatives used were Boc-Cys(Mob)-OH, Boc-Phe-OH, Boc-Ile-OH, Boc-Asp(OFm)-OH, Boc-Asn-OH, Boc-Pro-OH, Boc-Dbu(benzyloxycarbonyl)-OH DCHA salt and Boc-Gly-OH, all purchased from Novabiochem and Bachem.. HPLC and MS operations were performed as in the synthesis of compound 1. The fully protected peptide resin was manually synthesiseti starting from 0.6 g (0.4 mmol) of 4-methyl-benzhydrylamine resin (Novabiochem). DCC, PyBOP or DIC/HOBt mediated single couplings with 2.5-fold excess of amino acid derivatives were employed. The Boc group was removed with 50% TFA in DCM containing 1% of m-cresol. Upon completion of the synthesis/ the 9-fluoren-ylmethyl ester was removed from the p-carboxylic group of aspartic acid by treatment with 30% piperidine in DKF for 2 x 30 min. The resin was washed with 1 M HOBt in DMF solution for 30 rain and then twice with DMF only. The free carbcxylic group was amidated by overnight trea-ment with 2 mmcl of ethylamine/PyBOP/DIEA in DMF. The finished resin was washed with methanol, DMF and DCM and dried in vacua. The peptide was cleaved from the resin by using 30 ml of anhydrous HF containing 3 ml of anisole at 0°C for 90 minutes. The HF was evaporated off, and the crude linear pep-ide was washed with diethyl ether. The peptide was immediately dissolved in 200 nl of 25% acetonitrile/10% TFA (aq) and oxidised as described supra. The resulting mixture was loaded directly onto an HPLC column and purified using triethylammonium phosphate buffer at pH 2.3. The subsequent purification steps were identical to the procedure for compound 1^ 41 mg (-10% yield) of white amorphous powder was obtained. HPLC: Rt=10.0 min, gradient.: 20-40% B over 20 min, t=40°C, solvent A 0.01% TFA (aq), solvent B 70% CH3CN, 0.01% TFA (ag) ; Purity: 100%; MS (M-t-H4) : expected 992,5, observed 992.2. The other compounds were prepared by analogous variation of these synthetic procedures. ^xperimer.tal (biological testing) In vritro receptor assays: Agonist activity of compounds on the hVla receptor was determined in a transcriptional reporter assay by transiently transfecting a hVla receptor expression DNA into HEK-293 cells in concert with a reporter DNA containing intracellular calcium responsive promoter elements regulating expression of firefly luciferase. See Boss, V., Talpade, D.J., Murphy, T.J. J. Bid. Chem. 1996, May 3; 271(18), 10429-10432 for further guidance on this assay. Cells were exposed to serial dilutions of compounds diluted 10-fold per dose for 5 hours, followed by lysis of cells, determination of luciferace activity, and determination of compound efficacies and ECso values through non-linear regression. Arginine-vasopressin (AVP) was used as an internal control in each experiment, ana compounds were tested in at least three independent experiments. To determine selectivity, compounds were tested in luciferase-based tr&r.scriptional reporter assays expressing the human oxytocin (hOT) receptor. Assays for other receptors (hV2, hVlb, rat Via and rat V2) were also conducted. For further comparative purposes, other reference compounds used were [?he2,Qrn8]OT, terlipressin and 7180. The structure of [?he2,OrnB]OT is (SEQ ID NO: 51} : H-Cys-?he-Ile-Gln-Asn-Cys-Pro-Orn-Gly-NH2 T The structure of F180 is (SEQ ID NO: 52): Hmp-Phe-Ile-Bgn-Asn-Cys-Pro-Dbu(Abu)-Gly-NH2 The results of the in vitro assays are depicted in table 2 inr"ra. The EC5o value given is the geometric mean expressed in nanomol/L (nM). Selectivity values are given as ECso ratios. In vivo pharmacological tests: The compounds were tested in vivo for duration of action related to a standard dose of AVP. Blood pressure tests were carried out on anaesthetised Sprague-Dawley male rats (weighing 270-300 g) with catheterised jugular vein and carotid artery. The catheterised carotid artery WES used to continuously monitor blood pressure and the jugular vein was used for administration of the compounds tested. Rats received intravenous injections of dibenair.ine prior to dosing to enhance their responsiveness to Via receptor agonists (cf. Dekanski, J., Br. J. Pharmacol. 1952, 1, 567-572). The dosing procedure consisted of one intravenous injection of physiological saline followed by two consecutive injections of a standard dose of AVP (0.1 nmol/kg, * ED7o) , and three to five increasing doses of a giver, compound selected to give at least a response comparable to the standard dose of AVP. Dosing intervals were set as time for the blood pressure to decrease to a stable baseline. Determination of duration of action was based on the decay rate of diastolic arterial blood pressure transient increase. Specifically, for an exponential decay of plasma concentration, it can be shown that, if the response is measured beyond the distribution phase, the rate of decay near the ECso is linear and inversely proportional to the elimination half-life (Rowland, K. ani Tczer, T. in vClinical Pbzrmacokinetics, Concepts and Applications", 3rd ed., Lippincott Williams & Wilkins, Philadelphia, 1995}, To measure the response decay rate for a given compound, a dose was selected that gave an amplitude of response as similar as possible to the amplitude of response to the second injection of the standard dose of AVP. To normalise for inter-individual variation in Vla-responsiveness, the duration of action was expressed as the ratio of decay rate for this reference AVP response to the decay rate for the eguieffective dose of compound for each rat tested. The results obtained for the compounds tested are set forth in table 2. Table 2. Results of biological testing (Table Remove) (Table Remove) - = not tested •f = selective hVla receptor agonist; ECso hOT/r.Vla ratio not determined due to very low agonist efficacy ( All references listed are to be regarded as ar. integral part of the present writ. CLAIMS 1. A compound having the formula (I) (SEQ ID NO 53) : (Figure Remove) wherein: Ar is an aryl group selected from aromatic carbocyclic ring systems, five- or six-meinbered he^eroaromatic ring systems and bicyclic heteroaromatic ring systems; IR is selected from I, 2 and 3; n is selected from 0, 1, 2, 3 and 4; p is selected from 2, 3 and 4; Rir R2 and RS are independently selected from H, OH, alkyl, 0-alkyl and OC(0)-alkyl; alkyl is selected from Ci_e straight and C4_a branched chain alkyl and optionally has at least one hydroxyl substituent; and when n=0, RI and Ra optionally together form a nitrogen containing ring structure comprising from 2 to 5 carbon atoms; with the proviso that when Ar is phenyl, m=2, n=0, Ri=R2=H, ana p is 3 or 4 RS is not H; and solvates and pharmaceutically acceptable salts thereof. 2. A compound according to claim 1, wherein Ar is selected from phenyl, 2- or 3-thienyl, 2- or 3-furyl, 2-, 3- or 4-pyridyl and 2-, 4- or 5-thiazolyl. 3, A compound according to any one of claims 1-2, wherein RI is H. .4- A compound according to any one of claims 1-3, wherein p is 2 or 3. 5. A compound according to any one of claims 1-4, wherein R2 is selected from H, OH, CH3, CH2CH3, CH(CHs):f CH{CH2OH)2, CK(OH)CH3, OCH3 and OCH2CH2OH. 6. A compound according to any one of claims 1-5, wherein RS is selected from H, methyl, ethyl, n-propyl, i-propyl and i-amyl. 7. A compound according to any one of claims 1-6, wherein the compound is selected from a group consisting of (SEQ ID NCs: 1-7, respectively, in order of appear ance) : H-Cys-Phe-IIe-Hgn-Asn-Cys-Pro-Orn(i-Pr)-Gly-NH2 r H-Cys-Phe-Ile-Asn ( (CH2) 3OH) -Asn-Cys-Pro-Orn-Gly-NK2 H-Cys-Phe-Iie-Asn-Asn-Cys-Pro-Dbu-Gly-NH2 K-Cys-Phe-lle-Asn (C.H2CH3) -Asn-Cys-Pro-Dbu-Gly-NH2 H-Cys-Phe-Iie-Gln-Asn-Cys-Pro-Orn(1-Pr)-Gly-NH2 and H-Cys-Phe-Ile-Gln-Asn-Cys-Pro-Orn (CH2CH3) -Gly-NH2. 8. A compound according to claim 1 having the structural formula: H-Cys~Phe-Ile-Asn(CH3)2-Asn-Cvs-Pro-Orn-Gly-NH2 9. A compound according to any one of claims 1-8 for use as a pharmaceutical- 10. A pharmaceutical composition comprising a compound according to any one of claims 1-8 as active ingredient in association with a pharmaceutically acceptable adjuvant, diluent or carrier.. 11. Use of a compound according to any one of claims 1-8 for the manufacture of a medicament for treatment of shock of hypovolemic or vasodilatory origin, bleeding esophageal varices, hepatorenal syndrome, cardiopulmonary resuscitation, anesthesia-induced hypotension, ortho- static hypotension, paracentesis-induced circulatory dys function, intra-operative blood loss or blood loss asso ciated with burr, debridement and epistaxis, and for treatment of various ocular diseases by increasing lacri- mstion/tear formation. 12. A method for treatment of shock of hypovolerr.ic or vasodilatory origin, bleeding esophageal varices, hepatorenal syndrome, cardiopulmonary resuscitation, an esthesia-induced hypotension, orthostatic hypotension, paracentesis-induced circulatory dysfunction, intra- operative blood loss or blood loss associated with burr. debridement and epistaxis, and of various ocular diseases by increasing lacrimation/tear formation, wherein said method comprises administering to an animal, including human, patient of a therapeutically effective amount of a compound according to any one of claims 1-8. cantPEPTIDIC VASOPRESSIN RECEPTOR AGONISTS Field of the Invention The present invention relates to novel compounds, pharmaceutical compositions comprising the same, use of said compounds for the manufacture of a medicament for treatment of inter alia shock conditions as well as to a method for treatment of said conditions, wherein said compounds Ere administered. Background Peptidic vasopressin Via receptor agonists, such as terlipressir., have recently (see e.g. O'Brian et al. , Lancet 359 ; £313):1209-10, June 4th, 2002) received increased attention for clinical use in treatment of critical care diseases and conditions, including shock-of hypovolemic (e.g. hemorrhagic) or vasodilatory (e.g. septic) origin, bleeding esophageal varices (BEV), hepa-torenal syndrome (HRS), cardiopulmonary resuscitation and anesthesia-induced hypotension. They have also beer, shown to have clinical use in the treatment of orthostatic hypotension, paracentesis-induced circulatory dysfunction, intra-opexative blood loss and blood loss associated with burn debridement and epistaxis, and for treatment of various ocular diseases by increasing lacrimation/tear formation. In treating critical care conditions it is highly desirable to control the arterial blood pressure, and the drug used is typically administered intravenously.. Continuous intravenous drug infusion at increasing or decreasing rates is a practical means of providing the desired degree of control. The attainment of so-called "steady state" plasma concentrations of drug depends on the elimination half life of the drug infused. It is generally recognised that steady state plasma concentration is achieved after a period of time equivalent to three times the elimination half life of the drug. To be prac- tical in a clinical setting the desired arterial blooci pressure at the steady state should be attained in about two hours, preferably in one hour or less. Via agonists with an elimination half life longer than 1 hour are therefore usually not considered useful for critical care treatment, A disadvantage of terlipressin in many critical care situations is its long duration of action, which makes it difficult to titrate its effect as the disease state changes. The efficacy of terlipressin at the human Via (hVia) receptor also needs to be improved e.g. to allow lower dosages in general. Also the compound known as F180 {cf. example 3 in US patent No. 5,459,236) has an inconveniently long duration of action to be considered for the treatment of most critical care conditions. Non-specific receptor agonist activity is the main disadvantage of other existing compounds, e.g. [Phe2,Orn8jOT (cf. example If in US patent No. 3,352,843) and arginine-vasopressin (AVP). Activity at related receptors such as Vlb, V.2 and oxytocin (OT) receptors may potentially generate undesirable side effects and safety concerns. As an example, V2 receptor activation may induce antidiuresis (cf. desmopressrn), release of coagulation/thromboly sis factors, and induce vasodilation/hypo-tension with reflex tachycardia. The latter side effect may also be induced by OT receptor agonist activity. It is an objective of the present invention to provide compounds that are especially useful in the treatment of critical care conditions. Disclosure of the Invention The present invention relates to compounds represented by the general formula (I) (SEQ ID NO RaN(CH2)nR2 CH2 H-Cys^ A-xIle. Is,Asn-Cys-Prox ^X^Gly-NHo (I H T E T N T o H o H wherein: Ar is an aryl group selected from aromatic carbocyclic ring systems, five- or six-membereci heteroaronaatic ring systems and bicyclic heteroaromatic ring systems; m is selected from 1, 2 and 3; n is selected from 0, 1, 2, 3 and 4; p is selected from 2, 3 and 4; Ri, R2 and Rs are independently selected from H, OK, alkyl, 0-alkyI and OC(0)-alkyl; alkyl is selected from Ci_6 straight and C^-E branched chain alkyl and optionally has at least one hydroxyl substituent; and when n«0, HI and R2 optionally together form a nitrogen containing ring structure comprising from 2 to 5 carbon atoms; with the proviso that when Ar is phenyl (amino acid no, 2 is Phe), m«=2r n=0 and H!=R2=fl (amirio acid no.. 4 is Gin) R3 is riot H when p is 3 or 4; and solvates and pharmaceutically acceptable salts thereof. Amino acid no. B is Ozn when R3=H and p=3, and Lys when R3=H and p=4. For the purposes of the present invention, the following terminology is used-Aromatic carbocyclic ring systems includes phenyl and xiaphthyl. A five-membered heteroaromatic ring system is a monocyclic aromatic ring system having five ring atoms, wherein 1, 2 or 3 ring atoms are independently selected from N, 0 and S. Preferred such ring systems are selected form a group consisting of thienyl, furyl, pyrrolyl, imidazolyl, thiazolyl, oxazolyl, pyrazolyl, isothiazolyl, isoxazolyl and tetrazolyl. A six-membered heteroaromatic ring system is a mono-cyclic aromatic ring system having six ring atoms, wherein 1, 2 cr 3 ring atoms are independently selected from N, 0 and S. It is preferably selected from a group consisting of pyridyl, pyrazinyl, pyrimidinyl, triazir.yl and pyridazinyl. A bicyclic heteroaromatic ring system is a ring system having two five- or six-membered heteroaromatic rings, or a phenyl and a five- or six-membered heteroaxo-matic ring, cr a phenyl and a hererocyclyl ring, or a five- or six-membered heteroaromatic ring and a heterocy-clyl ring; connected by a ring fusion, said bicyclic heteroaromatic ring system comprising 8 to 12 ring atoms, wherein 1, 2 or 3 of the ring a~oms are independently selected from N, 0 and S, It is preferably selected from a group consisting of indole, guinoline, tetrahydroguino-line, isoquinoline, tetrahydroisoguinoline, 1,4-benzo-dioxan, coumarin, benzofuran, Ir2-benzisoxazole, ben-zothiophene, benzoxazole, benzthiazole, benzimidazole, benzxriazole, pyrolizidine and quinoliziditie, A heterocyclyl or heterocyclic moiety is a saturated or partially saturated ring system having 3 to 7 ring atoms, wherein 1, 2 or 3 ring atoms are independently selected from N, 0 and S, Heterocyclyl moieties are preferably selected from a group consisting of aziridine, oxirane, thiirane, azetidine, oxetane, thietane, pyr-rolidine., pyrroline, imidazolidine, pyrazolidine, di-oxolaner tetrahydrofuranylf piperidine, pipexazine, mor-pholine(/ tet:rahydropyranylf 1,4-dioxanyl, homopiperid-homopiperazinyl and hexamethylene oxide. It deserves mentioning that e.g. also isopropyl and 2-rt-butyl groups are encompassed by the expression Ci_6 straight chain alkyl, as said expression is not related to the binding site of the straight chain in question. Ci_€ denotes having from one to six carbon atoms, including any number therebetween, and this nomenclature is used analogously herein. Examples of phermaceurically acceptable salts comprise acid addition salts, e.g. a salt formed by reaction wirh hydrohalogen acids, such as hydrochloric acid, and mineral acids, such as sulphuric acid, phosphoric acid and nitric acid, as well as aliphatic, alicyclic, aromatic or heterocyclic suiphonic or carboxylic acids, sucft as formic acid, acetic acid, propionic acid, succinic acid, glycolic acid, lactic. acid, malic acid, tartaric acid, citric acid, ascorbic acid, maleic acid, hydroxymaleic acid, pyruvic acid, p-hydroxybenzcic acid, embonic acid, methanesulphonic acid, ethanesulphonic acid, hydroxyethanesulphonic acid, halobenzenesulphonic acid, toluenesulphonic acid and naphtalenesulphonic acid. Ar is preferably selected from phenyl, 2- or 3-thier.yl, 2- or 3-furyl, 2-, 3- or 4-pyridyl and 2-, 4- or 5-thiazolyl, It is particularly preferred that RI is H. In preferred embodiments p is 2 or 3.. It is preferred to select R2 from H, OH, CH3, CH2CH3, CH{CH3)2, CEiCH2OH)2, CH(OH)CH3 (both enantiomers) , OCH3 and OCH2CH2OH. Moreover, it is preferred to select R3 from H, methyl, ethyl, n-propyl, i-propyl and i-amyl. In the most preferred embodiment, said compound having the formula (I) is selected from a group consisting of (SEQ ID NOs: 1-7, respectively, in order of appear- H-Cys-Phe-Ile-Hgn-Asn-Cys-Pro-Orn(i-Pr)-Gly-NH2 y H-Cys-Phe-Ile-Asn { (CH2) 3OH) -Asn-Cys-Pro-Orn-Gly-NH;, (2) H-Cys-Phe-Ile-Asn-Asn~Cys-Pro-Dbu-Gly-NH2 H-Cys-Phe-Ile-Asn (CH2CH3) -Asn-Cys-Pro-Dbu-Gly-NH2 I _ I (4 ) H-Cys-Phe-Ile-Gln-Asn-Cys-Pro-Orn (i-Pr) -Gly-NH2 and H-Cys-Phe-Ile-Gln-Asn-Cys-Pro-Orn(CH2CH3) -Gly-NH2, I I :6> H-Cys-Phe-Iie-Asn(CH3}2-Asn-Cys-Pro-Orn-Gly-NH2 I J (7) The number in parenthesis denotes the compound as referered to in the following. Furthermore the present invention relates to a compound as set forth above for use as a pharmaceutical. Accordingly, the present invention also relates to a pharmaceutical composition comprising a compound as set forth above as active ingredient in association with a pharmaceutically acceptable adjuvant, diluent or carrier. The pharmaceutical composition may be adapted for oral, intravenous, topical, intraperitoneal, nasal, buccal, sublingual or subcutaneous administration cr for administration via the respiratory tract e.g. in the. form of an aerosol or an air-suspended fine powder. The composition may thus for instance be in the form of tablets, capsules., powders, microparticles, granules, syrups, suspensions, solutions, transdermal patches or suppositories. It .should be noted that the composition according to the present invention may optionally include two or more of the above outlined compounds. The present pharmaceutical composition may optionally comprise e.g. at least one further additive selected from a disintegrating agent, binder, lubricant, flavoring agent, preservative, colorant and any mixture thereof. Examples of such and other additives are found in "%Handbook of Pharmaceutical Excipients"; Ed. A.H. Kibbe, 3rd Ed., American Pharmaceutical Association, USA and Pharmaceutical Press UK, 2000. The present pharmaceutical composition is most preferably adapted for parenteral administration. It may comprise a sterile aqueous preparation of the compounds of the invention preferably isotonic with the blood of the recipient. This aqueous preparation may be formulated according 10 kn.own methods using suitable dispersing or wetting agents and suspending agents. The injectable aqueous formulation Remestyp® (terlipressin) is exemplary of a suitable pharmaceutical formulation. The preparation may also be a sterile injectable solution or suspension in a diluent or solvent, for example as a solution in 1,3-butane diol. Water, Ringer's solution, and isotonic sodium chloride solution are exemplary acceptable diluents.. Sterile, fixed oils may be employed as a solvent or suspending medium. Bland fixed oils, including synthetic mono or di-glycerides, and fatty acids, such as oleic acid, may also be used. In addition, the present invention relates tc use of a compound as outlined above for the manufacture of a medicament for treatment of shock of hypovolemic or vasodi-latory origin, BEV, HRS, cardiopulmonary resuscitation, anesthesia-induced hypotension, orthostatic hypotension, paracentesis-induced circulatory dysfunction, intra-operative blood loss or blood loss associated with burn debridement and epistaxis, and for treatment of various ocular diseases by increasing lacrimation/tear formation. In another embodiment the invention relates to a method for treatment of shock of hypovolemic or vasodila-tory origin., BEV, .HRS, cardiopulmonary resuscitation, an- esthesia-induced hypotension, orthostatic hypotension, paracentesis-induced circulatory dysfunction, intra-operative blood loss or blood loss associated with burn debridement and epistaxis, and of various ocular diseases by increasing lacrimation/tear formation, wherein said method comprises administering to an animal, including human, patient of a therapeutically effective amount of a compound as outlined above. The typical dosage of the compounds according to the present invention varies within a wide range and will depend on various factors such as the individual needs of each patient and the route of administration. The dosage administered by infusion is generally within the range of 0.01-200 ug/kg body weight per hour. A physician of ordinary skill in the art will be able to optimise the dosage to the situation at hand. The abbreviations used are: Abu 2-aminobutyric acid Boc tert-butoxycarbonyl BOP benzotriazol-1-yloxy "trisdimethylamino- phosphonium hexafluorophosphate Dbu 2,4-diaminobutyric acid DCC N,N'-dicyclohexylcarbodiimide DCHA dicyclohexylamine DCM dichloromethane DIAD diisopxopyl diazodicarboxylate DIG W/W-diisopropylcarbodiimide DIEA N, N-diisopropyl-JV-ethylamine DMF Nr jW-dimethylformamide Fm 9-fluorenylmethyl Fmoc S-fluorenylmethoxycarbonyl Hgn homoglutamine Hmp 2-hydroxy-3-mercaptopropionic acid HOBt 1-hydroxybenzotriazole HPLC high performance liquid chromatography i iso Mint 4-methoxytrityl Mob p-methoxybenzyl MS mass spectrometry Orn ornithine Ph phenyl Pr propyl PyBOP benzotriazol-1-yloxy trispyrrolidine- phosphonium hexafluorophosphate o-NBS-Cl2-nitrobenzenesulfonyl chloride OT oxytocin Rt retention time TFA trifluoroacetic acid TIS triisopropylsilane TMOF trimethylorthoformate TPP triphenylphosphine Trt trityl VT vasotocin, [lie3] vasopressin Unless otherwise specified L-amino acids were used, and conventional amino acid terminology is adhered to. Experimental (synthesis) Amino acid derivatives and resins were purchased from commercial providers (Novabiochem, Bachem Peptide International and PepTech Corporation). Fmoc-Hgn-OH was synthesised according to literature (Wisniewski, K., Kolodziejczyk, A..S, Org.. Prep. P-roced. Int.. 1997, 29, 338-341). Other chemicals and solvents were provided from Sigma-Aldrich, Fisher Scientific and VWR. The compounds herein were synthesised by standard methods in solid phase peptide chemistry utilising both Fmoc and Boc methodology.. Unless otherwise provided, all reactions were performed at room temperature. In addition to the references cited supra, the following standard reference literature provides further guidance on general experimental set up, as well as on the availability of required starting material and reagents;: Kates, S.A., Albericio, F., Eds., Solid Phase Synthesis. A Practical Guide, Marcel Dekker, New York, Basel, 2000; Stewart, J.M., Young, J.D. Solid Phase Synthesis, Pierce Chemical Company, 1984; Bisello, et al., J. Biol. Chem, 199B, 273, 22496-22505; and Merrifield, J, Am. Chem, Soc. 1963, 85, 2149-2154. Purity of the synthesized peptide may be determined by analytical reversed phase HPLC. Structural integrity of the peptides may be confirmed using amino acid analysis and electrospray mass spectrometry. The peptides synthesised by Fmoc methodology were cleaved with a TFA/TIS/H20 96/2/2 (v/v/v) solution, and cleavage in Boc methodology was accomplished with 90% HF/10% aniscle (v/v) solution. Disulfide bridge (ring) formation was achieved by oxidation of linear peptides dissolved in 10% TFA (aq) with iodine. Peptides were purified by preparative H?LC in triethylammonium phosphate buffers (aq}. The compounds were finally converted to acetate salts using conventional HPLC methodology. The fractions with a purity exceeding 97% were pooled and lyophilised. Synthesis of peptides with alkylated side chain in position no, 8: The peptides were assembled with Fmoc methodology. The diamrno acid residue in position no, 8 was introduced with an acid labile (i.e. removable with a solution containing 1-2% TFA) protecting group, such as methoxy-trityl (Mmt; see Barlos, K. et al. in Peptides 1992, Schneider, C.H., Eberle, A.N., Eds., ESCOM Science Publishers 3-V.r 1993, pp 263-284). Resin bound peptide was treated with a DCM/TIS/TFA 93/5/2 (v/v/v) solution for the Mmt group removal. Reductive alkylation with acetone/NaBH(OAc)s provided the W-isopropyl peptide. To avoid undesirable N,N-d±alkylation in reductive alkylation in the above procedure, which .may occur when straight chain alkyl aldehydes are used, an alternative was developed, wherein after the Mmt removal the amino group was first derivatised with 2-nitrobenzenesulfonyl chloride (o-NBS-Cl; see Fukuyama, T.; Jow, C.-K.; Cheung, N. Tetrahedron Lett. 1995, 36, 6373-6374). The resulting sulphonamide was then alkylated with an appropriate alcohol under conventional Mitsunobu reaction conditions, typically utilising TPP/DIAD in 1,2-dimethoxyethane (Mitsunobu, 0. Synthesis 1981, 1-2-8) . The o-NBS-Cl group was subsequently removed with 5% potassium thiophenolate ±r. DMF, after which the peptide was cleaved from the resin. Synthesis of peptides with Ar-alkylated side chair, in position no. 4: The peptides were assembled with Boc methodology. The residue in position no. 4 was introduced in the sequence as 3oc-Asp(OFm)-OH. After complete peptide assembly the side chain protection was removed with 30% piperidine in DMF. The resulting free carboxylic group was converted to the desired amide by coupling with an appropriate amine mediated by PyBOP or BOP/DIEA. The K-terminal Boc group was then removed, followed by EF cleavage, cyclisation and purification by HPLC, Table 1 lists the compounds prepared by the above procedure, RI is H for all compounds except no- 7, where RI is CHa- An asterisk "" marks the most preferred embodiments. (Table Remove) 1. Compounds prepared with the formula (I\| (Table Remove) The following detailed examples are provided tc further illustrate the synthesis: Compound 1; [Phe2, Hgn4,0rn (i-Pr)e] VT: The amino acid derivatives used were Boc-Cys(Trt) -OH, Fmoc-Phe-OH, Fmoc-Ile-OH, Fmoc-Hgn-OH, Fmoc-Asn(Trt)-OK, Fraoc-CysETrt)-OHf Fmoc-Pro-OH, Fmoc-Orn(Mmt)-OH and Fmoc-Gly-OH. Fmoc-Hgn-OH was synthesised as mentioned above. Analytical HPLC was performed on a Waters 600 Liquid Chromatograph using a Vydac C18, 5u 4.6 x 250 mm, column at a flow rate of 2 ml/min. Preparative HPLC was performed on a Waters 2000 Liquid Chromatograph using a Prepak 47 x 300 mm cartridge at a flow rate of IOC ml/min, Final compound analysis was performed on a 1100 Agilent Liquid Chromatograph using a Vydac C18r 5ja 2,1 x 250 mm, column at a flow rate of 0.3 ml/min. Mass spectra were recorded on a Finnigan MAT spectrometer. The fully protected peptide resin was synthesised on an Applied Biosystems 9050 Peptide Synthesiser starting from 2 g (0.5 mmol) of Tentagel-S-RAM resin (Peptides International). DIC/HOBt mediated single couplings with a 4-fold excess of amino acid derivatives were performed. The Fmoc group was removed with 20% piperidine in DMF. Upon completion of the automated synthesis, the resin was transferred into a manual synthesis vessel and was treated with DCM/TIS/TFA 93/5/2 (v/v/v) solution (30 ml) for 2 x 1.5 hours for removal of the Mmt group. The resin was thoroughly washed with DCM and was subsequently suspended in 15 ml of 1,2-dichloroetehane/TMOF 1:1 (v/v) . 0.2 ml of acetone was then added followed by 0,6 g :;f NaBH(OAc)3. The suspension was shaken overnight and \:he resin was washed with methanol, DMF and DCM and dried in vacua. The resin was then treated with 30 ml of the TFA/TIS/%0 96/2/2 (v/v/v) solution for 1.5 hours and filtered off. The filtrate was evaporated and the crude linear peptide was precipitated with diethyl ether. The precipitate was immediately dissolved in 500 ml of 10% TFA (aq), and the peptide was oxidised by adding 0.1 M T2 in methanol to the magnetically stirred solution unzil yellow color persisted. Excess of iodine was reduced with ascorbic acid. The reaction mixture was then cooled with crushed ice and pH was adjusted to about 5 by'adding concentrated ammonia (aq) . The mixture was loaded onto an HPLC column and purified using a triethylammonium phosphate buffer with pH 5.2. The compound was eluted with a gradient of acetonitrile. The fractions with a purity exceeding 97% were pooled, and the resulting solution was diluted with 2 volumes of water. The solution was reloaded onto the column which was then washed with 2 1 of 0,1 M ammonium acetate (aq) and equilibrated with 2% acetic acid (aq). The compound was eluted with a fast (3%/min) gradient of acetonitrile. The fractions containing the desired product were pooled and lyophillsed.. 168 mg (-30% yield) of white amorphous powder was obtained. HPLC: Rt=8.5 min, gradient; 20-. Compound 4? [Phe2,Asn (Et)4, Dbu8]VT : The amino acid derivatives used were Boc-Cys(Mob)-OH, Boc-Phe-OH, Boc-Ile-OH, Boc-Asp(OFm)-OH, Boc-Asn-OH, Boc-Pro-OH, Boc-Dbu(benzyloxycarbonyl)-OH DCHA salt and Boc-Gly-OH, all purchased from Novabiochem and Bachem.. HPLC and MS operations were performed as in the synthesis of compound 1. The fully protected peptide resin was manually synthesiseti starting from 0.6 g (0.4 mmol) of 4-methyl-benzhydrylamine resin (Novabiochem). DCC, PyBOP or DIC/HOBt mediated single couplings with 2.5-fold excess of amino acid derivatives were employed. The Boc group was removed with 50% TFA in DCM containing 1% of m-cresol. Upon completion of the synthesis/ the 9-fluoren-ylmethyl ester was removed from the p-carboxylic group of aspartic acid by treatment with 30% piperidine in DKF for 2 x 30 min. The resin was washed with 1 M HOBt in DMF solution for 30 rain and then twice with DMF only. The free carbcxylic group was amidated by overnight trea-ment with 2 mmcl of ethylamine/PyBOP/DIEA in DMF. The finished resin was washed with methanol, DMF and DCM and dried in vacua. The peptide was cleaved from the resin by using 30 ml of anhydrous HF containing 3 ml of anisole at 0°C for 90 minutes. The HF was evaporated off, and the crude linear pep-ide was washed with diethyl ether. The peptide was immediately dissolved in 200 nl of 25% acetonitrile/10% TFA (aq) and oxidised as described supra. The resulting mixture was loaded directly onto an HPLC column and purified using triethylammonium phosphate buffer at pH 2.3. The subsequent purification steps were identical to the procedure for compound 1^ 41 mg (-10% yield) of white amorphous powder was obtained. HPLC: Rt=10.0 min, gradient.: 20-40% B over 20 min, t=40°C, solvent A 0.01% TFA (aq), solvent B 70% CH3CN, 0.01% TFA (ag) ; Purity: 100%; MS (M-t-H4) : expected 992,5, observed 992.2. The other compounds were prepared by analogous variation of these synthetic procedures. ^xperimer.tal (biological testing) In vritro receptor assays: Agonist activity of compounds on the hVla receptor was determined in a transcriptional reporter assay by transiently transfecting a hVla receptor expression DNA into HEK-293 cells in concert with a reporter DNA containing intracellular calcium responsive promoter elements regulating expression of firefly luciferase. See Boss, V., Talpade, D.J., Murphy, T.J. J. Bid. Chem. 1996, May 3; 271(18), 10429-10432 for further guidance on this assay. Cells were exposed to serial dilutions of compounds diluted 10-fold per dose for 5 hours, followed by lysis of cells, determination of luciferace activity, and determination of compound efficacies and ECso values through non-linear regression. Arginine-vasopressin (AVP) was used as an internal control in each experiment, ana compounds were tested in at least three independent experiments. To determine selectivity, compounds were tested in luciferase-based tr&r.scriptional reporter assays expressing the human oxytocin (hOT) receptor. Assays for other receptors (hV2, hVlb, rat Via and rat V2) were also conducted. For further comparative purposes, other reference compounds used were [?he2,Qrn8]OT, terlipressin and 7180. The structure of [?he2,OrnB]OT is (SEQ ID NO: 51} : H-Cys-?he-Ile-Gln-Asn-Cys-Pro-Orn-Gly-NH2 T The structure of F180 is (SEQ ID NO: 52): Hmp-Phe-Ile-Bgn-Asn-Cys-Pro-Dbu(Abu)-Gly-NH2 The results of the in vitro assays are depicted in table 2 inr"ra. The EC5o value given is the geometric mean expressed in nanomol/L (nM). Selectivity values are given as ECso ratios. In vivo pharmacological tests: The compounds were tested in vivo for duration of action related to a standard dose of AVP. Blood pressure tests were carried out on anaesthetised Sprague-Dawley male rats (weighing 270-300 g) with catheterised jugular vein and carotid artery. The catheterised carotid artery WES used to continuously monitor blood pressure and the jugular vein was used for administration of the compounds tested. Rats received intravenous injections of dibenair.ine prior to dosing to enhance their responsiveness to Via receptor agonists (cf. Dekanski, J., Br. J. Pharmacol. 1952, 1, 567-572). The dosing procedure consisted of one intravenous injection of physiological saline followed by two consecutive injections of a standard dose of AVP (0.1 nmol/kg, * ED7o) , and three to five increasing doses of a giver, compound selected to give at least a response comparable to the standard dose of AVP. Dosing intervals were set as time for the blood pressure to decrease to a stable baseline. Determination of duration of action was based on the decay rate of diastolic arterial blood pressure transient increase. Specifically, for an exponential decay of plasma concentration, it can be shown that, if the response is measured beyond the distribution phase, the rate of decay near the ECso is linear and inversely proportional to the elimination half-life (Rowland, K. ani Tczer, T. in vClinical Pbzrmacokinetics, Concepts and Applications", 3rd ed., Lippincott Williams & Wilkins, Philadelphia, 1995}, To measure the response decay rate for a given compound, a dose was selected that gave an amplitude of response as similar as possible to the amplitude of response to the second injection of the standard dose of AVP. To normalise for inter-individual variation in Vla-responsiveness, the duration of action was expressed as the ratio of decay rate for this reference AVP response to the decay rate for the eguieffective dose of compound for each rat tested. The results obtained for the compounds tested are set forth in table 2. Table 2. Results of biological testing (Table Remove) (Table Remove) - = not tested •f = selective hVla receptor agonist; ECso hOT/r.Vla ratio not determined due to very low agonist efficacy ( All references listed are to be regarded as ar. integral part of the present writ. CLAIMS 1. A compound having the formula (I) (SEQ ID NO 53) : (Figure Remove) wherein: Ar is an aryl group selected from aromatic carbocyclic ring systems, five- or six-meinbered he^eroaromatic ring systems and bicyclic heteroaromatic ring systems; IR is selected from I, 2 and 3; n is selected from 0, 1, 2, 3 and 4; p is selected from 2, 3 and 4; Rir R2 and RS are independently selected from H, OH, alkyl, 0-alkyl and OC(0)-alkyl; alkyl is selected from Ci_e straight and C4_a branched chain alkyl and optionally has at least one hydroxyl substituent; and when n=0, RI and Ra optionally together form a nitrogen containing ring structure comprising from 2 to 5 carbon atoms; with the proviso that when Ar is phenyl, m=2, n=0, Ri=R2=H, ana p is 3 or 4 RS is not H; and solvates and pharmaceutically acceptable salts thereof. 2. A compound according to claim 1, wherein Ar is selected from phenyl, 2- or 3-thienyl, 2- or 3-furyl, 2-, 3- or 4-pyridyl and 2-, 4- or 5-thiazolyl. 3, A compound according to any one of claims 1-2, wherein RI is H. .4- A compound according to any one of claims 1-3, wherein p is 2 or 3. 5. A compound according to any one of claims 1-4, wherein R2 is selected from H, OH, CH3, CH2CH3, CH(CHs):f CH{CH2OH)2, CK(OH)CH3, OCH3 and OCH2CH2OH. 6. A compound according to any one of claims 1-5, wherein RS is selected from H, methyl, ethyl, n-propyl, i-propyl and i-amyl. 7. A compound according to any one of claims 1-6, wherein the compound is selected from a group consisting of (SEQ ID NCs: 1-7, respectively, in order of appear ance) : H-Cys-Phe-IIe-Hgn-Asn-Cys-Pro-Orn(i-Pr)-Gly-NH2 r H-Cys-Phe-Ile-Asn ( (CH2) 3OH) -Asn-Cys-Pro-Orn-Gly-NK2 H-Cys-Phe-Iie-Asn-Asn-Cys-Pro-Dbu-Gly-NH2 K-Cys-Phe-lle-Asn (C.H2CH3) -Asn-Cys-Pro-Dbu-Gly-NH2 H-Cys-Phe-Iie-Gln-Asn-Cys-Pro-Orn(1-Pr)-Gly-NH2 and H-Cys-Phe-Ile-Gln-Asn-Cys-Pro-Orn (CH2CH3) -Gly-NH2. 8. A compound according to claim 1 having the structural formula: H-Cys~Phe-Ile-Asn(CH3)2-Asn-Cvs-Pro-Orn-Gly-NH2 9. A compound according to any one of claims 1-8 for use as a pharmaceutical- 10. A pharmaceutical composition comprising a compound according to any one of claims 1-8 as active ingredient in association with a pharmaceutically acceptable adjuvant, diluent or carrier.. 11. Use of a compound according to any one of claims 1-8 for the manufacture of a medicament for treatment of shock of hypovolemic or vasodilatory origin, bleeding esophageal varices, hepatorenal syndrome, cardiopulmonary resuscitation, anesthesia-induced hypotension, ortho- static hypotension, paracentesis-induced circulatory dys function, intra-operative blood loss or blood loss asso ciated with burr, debridement and epistaxis, and for treatment of various ocular diseases by increasing lacri- mstion/tear formation. 12. A method for treatment of shock of hypovolerr.ic or vasodilatory origin, bleeding esophageal varices, hepatorenal syndrome, cardiopulmonary resuscitation, an esthesia-induced hypotension, orthostatic hypotension, paracentesis-induced circulatory dysfunction, intra- operative blood loss or blood loss associated with burr. debridement and epistaxis, and of various ocular diseases by increasing lacrimation/tear formation, wherein said method comprises administering to an animal, including human, patient of a therapeutically effective amount of a compound according to any one of claims 1-8. cant

Full Text

PEPTIDIC VASOPRESSIN RECEPTOR AGONISTS
Field of the Invention
The present invention relates to novel compounds, pharmaceutical compositions comprising the same, use of said compounds for the manufacture of a medicament for treatment of inter alia shock conditions as well as to a method for treatment of said conditions, wherein said compounds Ere administered. Background
Peptidic vasopressin Via receptor agonists, such as terlipressir., have recently (see e.g. O'Brian et al. , Lancet 359 ; £313):1209-10, June 4th, 2002) received increased attention for clinical use in treatment of critical care diseases and conditions, including shock-of hypovolemic (e.g. hemorrhagic) or vasodilatory (e.g. septic) origin, bleeding esophageal varices (BEV), hepa-torenal syndrome (HRS), cardiopulmonary resuscitation and anesthesia-induced hypotension. They have also beer, shown to have clinical use in the treatment of orthostatic hypotension, paracentesis-induced circulatory dysfunction, intra-opexative blood loss and blood loss associated with burn debridement and epistaxis, and for treatment of various ocular diseases by increasing lacrimation/tear formation.
In treating critical care conditions it is highly desirable to control the arterial blood pressure, and the drug used is typically administered intravenously.. Continuous intravenous drug infusion at increasing or decreasing rates is a practical means of providing the desired degree of control. The attainment of so-called "steady state" plasma concentrations of drug depends on the elimination half life of the drug infused. It is generally recognised that steady state plasma concentration is achieved after a period of time equivalent to three times the elimination half life of the drug. To be prac-

tical in a clinical setting the desired arterial blooci pressure at the steady state should be attained in about two hours, preferably in one hour or less. Via agonists with an elimination half life longer than 1 hour are therefore usually not considered useful for critical care treatment,
A disadvantage of terlipressin in many critical care situations is its long duration of action, which makes it difficult to titrate its effect as the disease state changes. The efficacy of terlipressin at the human Via (hVia) receptor also needs to be improved e.g. to allow lower dosages in general.
Also the compound known as F180 {cf. example 3 in US patent No. 5,459,236) has an inconveniently long duration of action to be considered for the treatment of most critical care conditions.
Non-specific receptor agonist activity is the main disadvantage of other existing compounds, e.g. [Phe2,Orn8jOT (cf. example If in US patent No. 3,352,843) and arginine-vasopressin (AVP). Activity at related receptors such as Vlb, V.2 and oxytocin (OT) receptors may potentially generate undesirable side effects and safety concerns. As an example, V2 receptor activation may induce antidiuresis (cf. desmopressrn), release of coagulation/thromboly sis factors, and induce vasodilation/hypo-tension with reflex tachycardia. The latter side effect may also be induced by OT receptor agonist activity.
It is an objective of the present invention to provide compounds that are especially useful in the treatment of critical care conditions.
Disclosure of the Invention
The present invention relates to compounds represented by the general formula (I) (SEQ ID NO
RaN(CH2)nR2
CH2 H-Cys^ A-xIle. Is,Asn-Cys-Prox ^X^Gly-NHo (I
H T E T N T
o H o H
wherein:
Ar is an aryl group selected from aromatic carbocyclic
ring systems, five- or six-membereci heteroaronaatic ring
systems and bicyclic heteroaromatic ring systems;
m is selected from 1, 2 and 3;
n is selected from 0, 1, 2, 3 and 4;
p is selected from 2, 3 and 4;
Ri, R2 and Rs are independently selected from H, OK,
alkyl, 0-alkyI and OC(0)-alkyl;
alkyl is selected from Ci_6 straight and C^-E branched
chain alkyl and optionally has at least one hydroxyl
substituent;
and when n«0, HI and R2 optionally together form a
nitrogen containing ring structure comprising from 2 to 5
carbon atoms;
with the proviso that when Ar is phenyl (amino acid no, 2
is Phe), m«=2r n=0 and H!=R2=fl (amirio acid no.. 4 is Gin) R3
is riot H when p is 3 or 4; and
solvates and pharmaceutically acceptable salts thereof. Amino acid no. B is Ozn when R3=H and p=3, and Lys
when R3=H and p=4.
For the purposes of the present invention, the
following terminology is used-Aromatic carbocyclic ring systems includes phenyl
and xiaphthyl.
A five-membered heteroaromatic ring system is a monocyclic aromatic ring system having five ring atoms, wherein 1, 2 or 3 ring atoms are independently selected from N, 0 and S. Preferred such ring systems are selected form a group consisting of thienyl, furyl, pyrrolyl, imidazolyl, thiazolyl, oxazolyl, pyrazolyl, isothiazolyl, isoxazolyl and tetrazolyl.
A six-membered heteroaromatic ring system is a mono-cyclic aromatic ring system having six ring atoms, wherein 1, 2 cr 3 ring atoms are independently selected from N, 0 and S. It is preferably selected from a group consisting of pyridyl, pyrazinyl, pyrimidinyl, triazir.yl and pyridazinyl.
A bicyclic heteroaromatic ring system is a ring system having two five- or six-membered heteroaromatic rings, or a phenyl and a five- or six-membered heteroaxo-matic ring, cr a phenyl and a hererocyclyl ring, or a five- or six-membered heteroaromatic ring and a heterocy-clyl ring; connected by a ring fusion, said bicyclic heteroaromatic ring system comprising 8 to 12 ring atoms, wherein 1, 2 or 3 of the ring a~oms are independently selected from N, 0 and S, It is preferably selected from a group consisting of indole, guinoline, tetrahydroguino-line, isoquinoline, tetrahydroisoguinoline, 1,4-benzo-dioxan, coumarin, benzofuran, Ir2-benzisoxazole, ben-zothiophene, benzoxazole, benzthiazole, benzimidazole, benzxriazole, pyrolizidine and quinoliziditie,
A heterocyclyl or heterocyclic moiety is a saturated or partially saturated ring system having 3 to 7 ring atoms, wherein 1, 2 or 3 ring atoms are independently selected from N, 0 and S, Heterocyclyl moieties are preferably selected from a group consisting of aziridine, oxirane, thiirane, azetidine, oxetane, thietane, pyr-rolidine., pyrroline, imidazolidine, pyrazolidine, di-oxolaner tetrahydrofuranylf piperidine, pipexazine, mor-pholine(/ tet:rahydropyranylf 1,4-dioxanyl, homopiperid-homopiperazinyl and hexamethylene oxide.
It deserves mentioning that e.g. also isopropyl and 2-rt-butyl groups are encompassed by the expression Ci_6 straight chain alkyl, as said expression is not related to the binding site of the straight chain in question.
Ci_€ denotes having from one to six carbon atoms, including any number therebetween, and this nomenclature is used analogously herein.
Examples of phermaceurically acceptable salts comprise acid addition salts, e.g. a salt formed by reaction wirh hydrohalogen acids, such as hydrochloric acid, and mineral acids, such as sulphuric acid, phosphoric acid and nitric acid, as well as aliphatic, alicyclic, aromatic or heterocyclic suiphonic or carboxylic acids, sucft as formic acid, acetic acid, propionic acid, succinic acid, glycolic acid, lactic. acid, malic acid, tartaric acid, citric acid, ascorbic acid, maleic acid, hydroxymaleic acid, pyruvic acid, p-hydroxybenzcic acid, embonic acid, methanesulphonic acid, ethanesulphonic acid, hydroxyethanesulphonic acid, halobenzenesulphonic acid, toluenesulphonic acid and naphtalenesulphonic acid.
Ar is preferably selected from phenyl, 2- or 3-thier.yl, 2- or 3-furyl, 2-, 3- or 4-pyridyl and 2-, 4- or 5-thiazolyl, It is particularly preferred that RI is H.
In preferred embodiments p is 2 or 3..
It is preferred to select R2 from H, OH, CH3, CH2CH3, CH{CH3)2, CEiCH2OH)2, CH(OH)CH3 (both enantiomers) , OCH3 and OCH2CH2OH.
Moreover, it is preferred to select R3 from H, methyl, ethyl, n-propyl, i-propyl and i-amyl.
In the most preferred embodiment, said compound having the formula (I) is selected from a group consisting of (SEQ ID NOs: 1-7, respectively, in order of appear-

H-Cys-Phe-Ile-Hgn-Asn-Cys-Pro-Orn(i-Pr)-Gly-NH2
y
H-Cys-Phe-Ile-Asn { (CH2) 3OH) -Asn-Cys-Pro-Orn-Gly-NH;,
(2)

H-Cys-Phe-Ile-Asn-Asn~Cys-Pro-Dbu-Gly-NH2

H-Cys-Phe-Ile-Asn (CH2CH3) -Asn-Cys-Pro-Dbu-Gly-NH2 I _ I

(4 )

H-Cys-Phe-Ile-Gln-Asn-Cys-Pro-Orn (i-Pr) -Gly-NH2 and
H-Cys-Phe-Ile-Gln-Asn-Cys-Pro-Orn(CH2CH3) -Gly-NH2,
I I :6>
H-Cys-Phe-Iie-Asn(CH3}2-Asn-Cys-Pro-Orn-Gly-NH2
I J (7)
The number in parenthesis denotes the compound as referered to in the following.
Furthermore the present invention relates to a compound as set forth above for use as a pharmaceutical.
Accordingly, the present invention also relates to a pharmaceutical composition comprising a compound as set forth above as active ingredient in association with a pharmaceutically acceptable adjuvant, diluent or carrier.
The pharmaceutical composition may be adapted for oral, intravenous, topical, intraperitoneal, nasal, buccal, sublingual or subcutaneous administration cr for administration via the respiratory tract e.g. in the. form of an aerosol or an air-suspended fine powder. The composition may thus for instance be in the form of tablets, capsules., powders, microparticles, granules, syrups, suspensions, solutions, transdermal patches or suppositories.
It .should be noted that the composition according to the present invention may optionally include two or more of the above outlined compounds.

The present pharmaceutical composition may optionally comprise e.g. at least one further additive selected from a disintegrating agent, binder, lubricant, flavoring agent, preservative, colorant and any mixture thereof. Examples of such and other additives are found in "%Handbook of Pharmaceutical Excipients"; Ed. A.H. Kibbe, 3rd Ed., American Pharmaceutical Association, USA and Pharmaceutical Press UK, 2000.
The present pharmaceutical composition is most preferably adapted for parenteral administration. It may comprise a sterile aqueous preparation of the compounds of the invention preferably isotonic with the blood of the recipient. This aqueous preparation may be formulated according 10 kn.own methods using suitable dispersing or wetting agents and suspending agents. The injectable aqueous formulation Remestyp® (terlipressin) is exemplary of a suitable pharmaceutical formulation. The preparation may also be a sterile injectable solution or suspension in a diluent or solvent, for example as a solution in 1,3-butane diol. Water, Ringer's solution, and isotonic sodium chloride solution are exemplary acceptable diluents.. Sterile, fixed oils may be employed as a solvent or suspending medium. Bland fixed oils, including synthetic mono or di-glycerides, and fatty acids, such as oleic acid, may also be used.
In addition, the present invention relates tc use of a compound as outlined above for the manufacture of a medicament for treatment of shock of hypovolemic or vasodi-latory origin, BEV, HRS, cardiopulmonary resuscitation, anesthesia-induced hypotension, orthostatic hypotension, paracentesis-induced circulatory dysfunction, intra-operative blood loss or blood loss associated with burn debridement and epistaxis, and for treatment of various ocular diseases by increasing lacrimation/tear formation.
In another embodiment the invention relates to a method for treatment of shock of hypovolemic or vasodila-tory origin., BEV, .HRS, cardiopulmonary resuscitation, an-

esthesia-induced hypotension, orthostatic hypotension, paracentesis-induced circulatory dysfunction, intra-operative blood loss or blood loss associated with burn debridement and epistaxis, and of various ocular diseases by increasing lacrimation/tear formation, wherein said method comprises administering to an animal, including human, patient of a therapeutically effective amount of a compound as outlined above.
The typical dosage of the compounds according to the present invention varies within a wide range and will depend on various factors such as the individual needs of each patient and the route of administration. The dosage administered by infusion is generally within the range of 0.01-200 ug/kg body weight per hour. A physician of ordinary skill in the art will be able to optimise the dosage to the situation at hand.
The abbreviations used are:
Abu 2-aminobutyric acid
Boc tert-butoxycarbonyl
BOP benzotriazol-1-yloxy "trisdimethylamino-
phosphonium hexafluorophosphate
Dbu 2,4-diaminobutyric acid
DCC N,N'-dicyclohexylcarbodiimide
DCHA dicyclohexylamine
DCM dichloromethane
DIAD diisopxopyl diazodicarboxylate
DIG W/W-diisopropylcarbodiimide
DIEA N, N-diisopropyl-JV-ethylamine
DMF Nr jW-dimethylformamide
Fm 9-fluorenylmethyl
Fmoc S-fluorenylmethoxycarbonyl
Hgn homoglutamine
Hmp 2-hydroxy-3-mercaptopropionic acid
HOBt 1-hydroxybenzotriazole
HPLC high performance liquid chromatography
i iso

Mint 4-methoxytrityl
Mob p-methoxybenzyl
MS mass spectrometry
Orn ornithine
Ph phenyl
Pr propyl
PyBOP benzotriazol-1-yloxy trispyrrolidine-
phosphonium hexafluorophosphate o-NBS-Cl2-nitrobenzenesulfonyl chloride
OT oxytocin
Rt retention time
TFA trifluoroacetic acid
TIS triisopropylsilane
TMOF trimethylorthoformate
TPP triphenylphosphine
Trt trityl
VT vasotocin, [lie3] vasopressin
Unless otherwise specified L-amino acids were used, and conventional amino acid terminology is adhered to.
Experimental (synthesis)
Amino acid derivatives and resins were purchased from commercial providers (Novabiochem, Bachem Peptide International and PepTech Corporation). Fmoc-Hgn-OH was synthesised according to literature (Wisniewski, K., Kolodziejczyk, A..S, Org.. Prep. P-roced. Int.. 1997, 29, 338-341). Other chemicals and solvents were provided from Sigma-Aldrich, Fisher Scientific and VWR.
The compounds herein were synthesised by standard methods in solid phase peptide chemistry utilising both Fmoc and Boc methodology.. Unless otherwise provided, all reactions were performed at room temperature. In addition to the references cited supra, the following standard reference literature provides further guidance on general experimental set up, as well as on the availability of required starting material and reagents;:

Kates, S.A., Albericio, F., Eds., Solid Phase Synthesis. A Practical Guide, Marcel Dekker, New York, Basel, 2000;
Stewart, J.M., Young, J.D. Solid Phase Synthesis, Pierce Chemical Company, 1984;
Bisello, et al., J. Biol. Chem, 199B, 273, 22496-22505; and
Merrifield, J, Am. Chem, Soc. 1963, 85, 2149-2154.
Purity of the synthesized peptide may be determined by analytical reversed phase HPLC. Structural integrity of the peptides may be confirmed using amino acid analysis and electrospray mass spectrometry.
The peptides synthesised by Fmoc methodology were cleaved with a TFA/TIS/H20 96/2/2 (v/v/v) solution, and cleavage in Boc methodology was accomplished with 90% HF/10% aniscle (v/v) solution. Disulfide bridge (ring) formation was achieved by oxidation of linear peptides dissolved in 10% TFA (aq) with iodine. Peptides were purified by preparative H?LC in triethylammonium phosphate buffers (aq}. The compounds were finally converted to acetate salts using conventional HPLC methodology. The fractions with a purity exceeding 97% were pooled and lyophilised.
Synthesis of peptides with alkylated side chain in position no, 8:
The peptides were assembled with Fmoc methodology. The diamrno acid residue in position no, 8 was introduced with an acid labile (i.e. removable with a solution containing 1-2% TFA) protecting group, such as methoxy-trityl (Mmt; see Barlos, K. et al. in Peptides 1992, Schneider, C.H., Eberle, A.N., Eds., ESCOM Science Publishers 3-V.r 1993, pp 263-284). Resin bound peptide was treated with a DCM/TIS/TFA 93/5/2 (v/v/v) solution for the Mmt group removal. Reductive alkylation with acetone/NaBH(OAc)s provided the W-isopropyl peptide.
To avoid undesirable N,N-d±alkylation in reductive alkylation in the above procedure, which .may occur when

straight chain alkyl aldehydes are used, an alternative was developed, wherein after the Mmt removal the amino group was first derivatised with 2-nitrobenzenesulfonyl chloride (o-NBS-Cl; see Fukuyama, T.; Jow, C.-K.; Cheung, N. Tetrahedron Lett. 1995, 36, 6373-6374). The resulting sulphonamide was then alkylated with an appropriate alcohol under conventional Mitsunobu reaction conditions, typically utilising TPP/DIAD in 1,2-dimethoxyethane (Mitsunobu, 0. Synthesis 1981, 1-2-8) . The o-NBS-Cl group was subsequently removed with 5% potassium thiophenolate ±r. DMF, after which the peptide was cleaved from the resin.
Synthesis of peptides with Ar-alkylated side chair, in position no. 4:
The peptides were assembled with Boc methodology. The residue in position no. 4 was introduced in the sequence as 3oc-Asp(OFm)-OH. After complete peptide assembly the side chain protection was removed with 30% piperidine in DMF. The resulting free carboxylic group was converted to the desired amide by coupling with an appropriate amine mediated by PyBOP or BOP/DIEA. The K-terminal Boc group was then removed, followed by EF cleavage, cyclisation and purification by HPLC,
Table 1 lists the compounds prepared by the above procedure, RI is H for all compounds except no- 7, where RI is CHa- An asterisk "*" marks the most preferred embodiments.
(Table Remove) 1. Compounds prepared with the formula (I|
(Table Remove)
The following detailed examples are provided tc further illustrate the synthesis:
Compound 1; [Phe2, Hgn4,0rn (i-Pr)e] VT:
The amino acid derivatives used were Boc-Cys(Trt) -OH, Fmoc-Phe-OH, Fmoc-Ile-OH, Fmoc-Hgn-OH, Fmoc-Asn(Trt)-OK, Fraoc-CysETrt)-OHf Fmoc-Pro-OH, Fmoc-Orn(Mmt)-OH and Fmoc-Gly-OH. Fmoc-Hgn-OH was synthesised as mentioned above. Analytical HPLC was performed on a Waters 600 Liquid Chromatograph using a Vydac C18, 5u 4.6 x 250 mm, column at a flow rate of 2 ml/min. Preparative HPLC was performed on a Waters 2000 Liquid Chromatograph using a Prepak 47 x 300 mm cartridge at a flow rate of IOC ml/min, Final compound analysis was performed on a 1100 Agilent Liquid Chromatograph using a Vydac C18r 5ja 2,1 x 250 mm, column at a flow rate of 0.3 ml/min. Mass spectra were recorded on a Finnigan MAT spectrometer.
The fully protected peptide resin was synthesised on an Applied Biosystems 9050 Peptide Synthesiser starting from 2 g (0.5 mmol) of Tentagel-S-RAM resin (Peptides International). DIC/HOBt mediated single couplings with a 4-fold excess of amino acid derivatives were performed. The Fmoc group was removed with 20% piperidine in DMF. Upon completion of the automated synthesis, the resin was transferred into a manual synthesis vessel and was treated with DCM/TIS/TFA 93/5/2 (v/v/v) solution (30 ml) for 2 x 1.5 hours for removal of the Mmt group. The resin was thoroughly washed with DCM and was subsequently
suspended in 15 ml of 1,2-dichloroetehane/TMOF 1:1 (v/v) . 0.2 ml of acetone was then added followed by 0,6 g :;f NaBH(OAc)3. The suspension was shaken overnight and \:he resin was washed with methanol, DMF and DCM and dried in vacua. The resin was then treated with 30 ml of the TFA/TIS/%0 96/2/2 (v/v/v) solution for 1.5 hours and filtered off. The filtrate was evaporated and the crude linear peptide was precipitated with diethyl ether. The precipitate was immediately dissolved in 500 ml of 10% TFA (aq), and the peptide was oxidised by adding 0.1 M T2 in methanol to the magnetically stirred solution unzil yellow color persisted. Excess of iodine was reduced with ascorbic acid. The reaction mixture was then cooled with crushed ice and pH was adjusted to about 5 by'adding concentrated ammonia (aq) . The mixture was loaded onto an HPLC column and purified using a triethylammonium phosphate buffer with pH 5.2. The compound was eluted with a gradient of acetonitrile. The fractions with a purity exceeding 97% were pooled, and the resulting solution was diluted with 2 volumes of water. The solution was reloaded onto the column which was then washed with 2 1 of 0,1 M ammonium acetate (aq) and equilibrated with 2% acetic acid (aq). The compound was eluted with a fast (3%/min) gradient of acetonitrile. The fractions containing the desired product were pooled and lyophillsed.. 168 mg (-30% yield) of white amorphous powder was obtained. HPLC: Rt=8.5 min, gradient; 20-. Compound 4? [Phe2,Asn (Et)4, Dbu8]VT :
The amino acid derivatives used were Boc-Cys(Mob)-OH, Boc-Phe-OH, Boc-Ile-OH, Boc-Asp(OFm)-OH, Boc-Asn-OH, Boc-Pro-OH, Boc-Dbu(benzyloxycarbonyl)-OH DCHA salt and Boc-Gly-OH, all purchased from Novabiochem and Bachem.. HPLC and MS operations were performed as in the synthesis of compound 1.
The fully protected peptide resin was manually synthesiseti starting from 0.6 g (0.4 mmol) of 4-methyl-benzhydrylamine resin (Novabiochem). DCC, PyBOP or DIC/HOBt mediated single couplings with 2.5-fold excess of amino acid derivatives were employed. The Boc group was removed with 50% TFA in DCM containing 1% of m-cresol. Upon completion of the synthesis/ the 9-fluoren-ylmethyl ester was removed from the p-carboxylic group of aspartic acid by treatment with 30% piperidine in DKF for 2 x 30 min. The resin was washed with 1 M HOBt in DMF solution for 30 rain and then twice with DMF only. The free carbcxylic group was amidated by overnight trea-ment with 2 mmcl of ethylamine/PyBOP/DIEA in DMF. The finished resin was washed with methanol, DMF and DCM and dried in vacua. The peptide was cleaved from the resin by using 30 ml of anhydrous HF containing 3 ml of anisole at 0°C for 90 minutes. The HF was evaporated off, and the crude linear pep-ide was washed with diethyl ether. The peptide was immediately dissolved in 200 nl of 25% acetonitrile/10% TFA (aq) and oxidised as described supra. The resulting mixture was loaded directly onto an HPLC column and purified using triethylammonium phosphate buffer at pH 2.3. The subsequent purification steps were identical to the procedure for compound 1^ 41 mg (-10% yield) of white amorphous powder was obtained. HPLC: Rt=10.0 min, gradient.: 20-*40% B over 20 min, t=40°C, solvent A 0.01% TFA (aq), solvent B 70% CH3CN, 0.01% TFA (ag) ; Purity: 100%; MS (M-t-H4) : expected 992,5, observed 992.2.
The other compounds were prepared by analogous variation of these synthetic procedures. ^xperimer.tal (biological testing)
In vritro receptor assays:
Agonist activity of compounds on the hVla receptor was determined in a transcriptional reporter assay by transiently transfecting a hVla receptor expression DNA into HEK-293 cells in concert with a reporter DNA
containing intracellular calcium responsive promoter elements regulating expression of firefly luciferase. See Boss, V., Talpade, D.J., Murphy, T.J. J. Bid. Chem. 1996, May 3; 271(18), 10429-10432 for further guidance on this assay. Cells were exposed to serial dilutions of compounds diluted 10-fold per dose for 5 hours, followed by lysis of cells, determination of luciferace activity, and determination of compound efficacies and ECso values through non-linear regression. Arginine-vasopressin (AVP) was used as an internal control in each experiment, ana compounds were tested in at least three independent experiments. To determine selectivity, compounds were tested in luciferase-based tr&r.scriptional reporter assays expressing the human oxytocin (hOT) receptor. Assays for other receptors (hV2, hVlb, rat Via and rat V2) were also conducted.
For further comparative purposes, other reference compounds used were [?he2,Qrn8]OT, terlipressin and 7180.
The structure of [?he2,OrnB]OT is (SEQ ID NO: 51} :
H-Cys-?he-Ile-Gln-Asn-Cys-Pro-Orn-Gly-NH2
T
The structure of F180 is (SEQ ID NO: 52): Hmp-Phe-Ile-Bgn-Asn-Cys-Pro-Dbu(Abu)-Gly-NH2
The results of the in vitro assays are depicted in table 2 inr"ra. The EC5o value given is the geometric mean expressed in nanomol/L (nM). Selectivity values are given as ECso ratios.
In vivo pharmacological tests:
The compounds were tested in vivo for duration of action related to a standard dose of AVP. Blood pressure tests were carried out on anaesthetised Sprague-Dawley male rats (weighing 270-300 g) with catheterised jugular vein and carotid artery. The catheterised carotid artery
WES used to continuously monitor blood pressure and the jugular vein was used for administration of the compounds tested. Rats received intravenous injections of dibenair.ine prior to dosing to enhance their responsiveness to Via receptor agonists (cf. Dekanski, J., Br. J. Pharmacol. 1952, 1, 567-572). The dosing procedure consisted of one intravenous injection of physiological saline followed by two consecutive injections of a standard dose of AVP (0.1 nmol/kg, * ED7o) , and three to five increasing doses of a giver, compound selected to give at least a response comparable to the standard dose of AVP. Dosing intervals were set as time for the blood pressure to decrease to a stable baseline.
Determination of duration of action was based on the decay rate of diastolic arterial blood pressure transient increase. Specifically, for an exponential decay of plasma concentration, it can be shown that, if the response is measured beyond the distribution phase, the rate of decay near the ECso is linear and inversely proportional to the elimination half-life (Rowland, K. ani Tczer, T. in vClinical Pbzrmacokinetics, Concepts and Applications", 3rd ed., Lippincott Williams & Wilkins, Philadelphia, 1995},
To measure the response decay rate for a given compound, a dose was selected that gave an amplitude of response as similar as possible to the amplitude of response to the second injection of the standard dose of AVP. To normalise for inter-individual variation in Vla-responsiveness, the duration of action was expressed as the ratio of decay rate for this reference AVP response to the decay rate for the eguieffective dose of compound for each rat tested. The results obtained for the compounds tested are set forth in table 2.
Table 2. Results of biological testing

(Table Remove)
(Table Remove)
- = not tested
•f = selective hVla receptor agonist; ECso hOT/r.Vla ratio not determined due to very low agonist efficacy ( All references listed are to be regarded as ar. integral part of the present writ.

CLAIMS
1. A compound having the formula (I) (SEQ ID NO
53) :

(Figure Remove)
wherein:
Ar is an aryl group selected from aromatic carbocyclic
ring systems, five- or six-meinbered he^eroaromatic ring
systems and bicyclic heteroaromatic ring systems;
IR is selected from I, 2 and 3;
n is selected from 0, 1, 2, 3 and 4;
p is selected from 2, 3 and 4;
Rir R2 and RS are independently selected from H, OH,
alkyl, 0-alkyl and OC(0)-alkyl;
alkyl is selected from Ci_e straight and C4_a branched
chain alkyl and optionally has at least one hydroxyl
substituent;
and when n=0, RI and Ra optionally together form a
nitrogen containing ring structure comprising from 2 to 5
carbon atoms;
with the proviso that when Ar is phenyl, m=2, n=0,
Ri=R2=H, ana p is 3 or 4 RS is not H; and
solvates and pharmaceutically acceptable salts thereof.
2. A compound according to claim 1, wherein Ar is
selected from phenyl, 2- or 3-thienyl, 2- or 3-furyl, 2-,
3- or 4-pyridyl and 2-, 4- or 5-thiazolyl.
3, A compound according to any one of claims 1-2,
wherein RI is H.
.4- A compound according to any one of claims 1-3, wherein p is 2 or 3.

5. A compound according to any one of claims 1-4,
wherein R2 is selected from H, OH, CH3, CH2CH3, CH(CHs):f
CH{CH2OH)2, CK(OH)CH3, OCH3 and OCH2CH2OH.
6. A compound according to any one of claims 1-5,
wherein RS is selected from H, methyl, ethyl, n-propyl,
i-propyl and i-amyl.
7. A compound according to any one of claims 1-6,
wherein the compound is selected from a group consisting
of (SEQ ID NCs: 1-7, respectively, in order of appear
ance) :
H-Cys-Phe-IIe-Hgn-Asn-Cys-Pro-Orn(i-Pr)-Gly-NH2

r
H-Cys-Phe-Ile-Asn ( (CH2) 3OH) -Asn-Cys-Pro-Orn-Gly-NK2

H-Cys-Phe-Iie-Asn-Asn-Cys-Pro-Dbu-Gly-NH2
K-Cys-Phe-lle-Asn (C.H2CH3) -Asn-Cys-Pro-Dbu-Gly-NH2
H-Cys-Phe-Iie-Gln-Asn-Cys-Pro-Orn(1-Pr)-Gly-NH2 and
H-Cys-Phe-Ile-Gln-Asn-Cys-Pro-Orn (CH2CH3) -Gly-NH2.
8. A compound according to claim 1 having the structural formula:
H-Cys~Phe-Ile-Asn(CH3)2-Asn-Cvs-Pro-Orn-Gly-NH2
9. A compound according to any one of claims 1-8 for
use as a pharmaceutical-
10. A pharmaceutical composition comprising a
compound according to any one of claims 1-8 as active
ingredient in association with a pharmaceutically
acceptable adjuvant, diluent or carrier..

11. Use of a compound according to any one of claims
1-8 for the manufacture of a medicament for treatment of
shock of hypovolemic or vasodilatory origin, bleeding
esophageal varices, hepatorenal syndrome, cardiopulmonary
resuscitation, anesthesia-induced hypotension, ortho-
static hypotension, paracentesis-induced circulatory dys
function, intra-operative blood loss or blood loss asso
ciated with burr, debridement and epistaxis, and for
treatment of various ocular diseases by increasing lacri-
mstion/tear formation.
12. A method for treatment of shock of hypovolerr.ic
or vasodilatory origin, bleeding esophageal varices,
hepatorenal syndrome, cardiopulmonary resuscitation, an
esthesia-induced hypotension, orthostatic hypotension,
paracentesis-induced circulatory dysfunction, intra-
operative blood loss or blood loss associated with burr.
debridement and epistaxis, and of various ocular diseases
by increasing lacrimation/tear formation, wherein said
method comprises administering to an animal, including
human, patient of a therapeutically effective amount of a
compound according to any one of claims 1-8.

cantPEPTIDIC VASOPRESSIN RECEPTOR AGONISTS
Field of the Invention
The present invention relates to novel compounds, pharmaceutical compositions comprising the same, use of said compounds for the manufacture of a medicament for treatment of inter alia shock conditions as well as to a method for treatment of said conditions, wherein said compounds Ere administered. Background
Peptidic vasopressin Via receptor agonists, such as terlipressir., have recently (see e.g. O'Brian et al. , Lancet 359 ; £313):1209-10, June 4th, 2002) received increased attention for clinical use in treatment of critical care diseases and conditions, including shock-of hypovolemic (e.g. hemorrhagic) or vasodilatory (e.g. septic) origin, bleeding esophageal varices (BEV), hepa-torenal syndrome (HRS), cardiopulmonary resuscitation and anesthesia-induced hypotension. They have also beer, shown to have clinical use in the treatment of orthostatic hypotension, paracentesis-induced circulatory dysfunction, intra-opexative blood loss and blood loss associated with burn debridement and epistaxis, and for treatment of various ocular diseases by increasing lacrimation/tear formation.
In treating critical care conditions it is highly desirable to control the arterial blood pressure, and the drug used is typically administered intravenously.. Continuous intravenous drug infusion at increasing or decreasing rates is a practical means of providing the desired degree of control. The attainment of so-called "steady state" plasma concentrations of drug depends on the elimination half life of the drug infused. It is generally recognised that steady state plasma concentration is achieved after a period of time equivalent to three times the elimination half life of the drug. To be prac-

tical in a clinical setting the desired arterial blooci pressure at the steady state should be attained in about two hours, preferably in one hour or less. Via agonists with an elimination half life longer than 1 hour are therefore usually not considered useful for critical care treatment,
A disadvantage of terlipressin in many critical care situations is its long duration of action, which makes it difficult to titrate its effect as the disease state changes. The efficacy of terlipressin at the human Via (hVia) receptor also needs to be improved e.g. to allow lower dosages in general.
Also the compound known as F180 {cf. example 3 in US patent No. 5,459,236) has an inconveniently long duration of action to be considered for the treatment of most critical care conditions.
Non-specific receptor agonist activity is the main disadvantage of other existing compounds, e.g. [Phe2,Orn8jOT (cf. example If in US patent No. 3,352,843) and arginine-vasopressin (AVP). Activity at related receptors such as Vlb, V.2 and oxytocin (OT) receptors may potentially generate undesirable side effects and safety concerns. As an example, V2 receptor activation may induce antidiuresis (cf. desmopressrn), release of coagulation/thromboly sis factors, and induce vasodilation/hypo-tension with reflex tachycardia. The latter side effect may also be induced by OT receptor agonist activity.
It is an objective of the present invention to provide compounds that are especially useful in the treatment of critical care conditions.
Disclosure of the Invention
The present invention relates to compounds represented by the general formula (I) (SEQ ID NO
RaN(CH2)nR2
CH2 H-Cys^ A-xIle. Is,Asn-Cys-Prox ^X^Gly-NHo (I
H T E T N T
o H o H
wherein:
Ar is an aryl group selected from aromatic carbocyclic
ring systems, five- or six-membereci heteroaronaatic ring
systems and bicyclic heteroaromatic ring systems;
m is selected from 1, 2 and 3;
n is selected from 0, 1, 2, 3 and 4;
p is selected from 2, 3 and 4;
Ri, R2 and Rs are independently selected from H, OK,
alkyl, 0-alkyI and OC(0)-alkyl;
alkyl is selected from Ci_6 straight and C^-E branched
chain alkyl and optionally has at least one hydroxyl
substituent;
and when n«0, HI and R2 optionally together form a
nitrogen containing ring structure comprising from 2 to 5
carbon atoms;
with the proviso that when Ar is phenyl (amino acid no, 2
is Phe), m«=2r n=0 and H!=R2=fl (amirio acid no.. 4 is Gin) R3
is riot H when p is 3 or 4; and
solvates and pharmaceutically acceptable salts thereof. Amino acid no. B is Ozn when R3=H and p=3, and Lys
when R3=H and p=4.
For the purposes of the present invention, the
following terminology is used-Aromatic carbocyclic ring systems includes phenyl
and xiaphthyl.
A five-membered heteroaromatic ring system is a monocyclic aromatic ring system having five ring atoms, wherein 1, 2 or 3 ring atoms are independently selected from N, 0 and S. Preferred such ring systems are selected form a group consisting of thienyl, furyl, pyrrolyl, imidazolyl, thiazolyl, oxazolyl, pyrazolyl, isothiazolyl, isoxazolyl and tetrazolyl.
A six-membered heteroaromatic ring system is a mono-cyclic aromatic ring system having six ring atoms, wherein 1, 2 cr 3 ring atoms are independently selected from N, 0 and S. It is preferably selected from a group consisting of pyridyl, pyrazinyl, pyrimidinyl, triazir.yl and pyridazinyl.
A bicyclic heteroaromatic ring system is a ring system having two five- or six-membered heteroaromatic rings, or a phenyl and a five- or six-membered heteroaxo-matic ring, cr a phenyl and a hererocyclyl ring, or a five- or six-membered heteroaromatic ring and a heterocy-clyl ring; connected by a ring fusion, said bicyclic heteroaromatic ring system comprising 8 to 12 ring atoms, wherein 1, 2 or 3 of the ring a~oms are independently selected from N, 0 and S, It is preferably selected from a group consisting of indole, guinoline, tetrahydroguino-line, isoquinoline, tetrahydroisoguinoline, 1,4-benzo-dioxan, coumarin, benzofuran, Ir2-benzisoxazole, ben-zothiophene, benzoxazole, benzthiazole, benzimidazole, benzxriazole, pyrolizidine and quinoliziditie,
A heterocyclyl or heterocyclic moiety is a saturated or partially saturated ring system having 3 to 7 ring atoms, wherein 1, 2 or 3 ring atoms are independently selected from N, 0 and S, Heterocyclyl moieties are preferably selected from a group consisting of aziridine, oxirane, thiirane, azetidine, oxetane, thietane, pyr-rolidine., pyrroline, imidazolidine, pyrazolidine, di-oxolaner tetrahydrofuranylf piperidine, pipexazine, mor-pholine(/ tet:rahydropyranylf 1,4-dioxanyl, homopiperid-homopiperazinyl and hexamethylene oxide.
It deserves mentioning that e.g. also isopropyl and 2-rt-butyl groups are encompassed by the expression Ci_6 straight chain alkyl, as said expression is not related to the binding site of the straight chain in question.
Ci_€ denotes having from one to six carbon atoms, including any number therebetween, and this nomenclature is used analogously herein.
Examples of phermaceurically acceptable salts comprise acid addition salts, e.g. a salt formed by reaction wirh hydrohalogen acids, such as hydrochloric acid, and mineral acids, such as sulphuric acid, phosphoric acid and nitric acid, as well as aliphatic, alicyclic, aromatic or heterocyclic suiphonic or carboxylic acids, sucft as formic acid, acetic acid, propionic acid, succinic acid, glycolic acid, lactic. acid, malic acid, tartaric acid, citric acid, ascorbic acid, maleic acid, hydroxymaleic acid, pyruvic acid, p-hydroxybenzcic acid, embonic acid, methanesulphonic acid, ethanesulphonic acid, hydroxyethanesulphonic acid, halobenzenesulphonic acid, toluenesulphonic acid and naphtalenesulphonic acid.
Ar is preferably selected from phenyl, 2- or 3-thier.yl, 2- or 3-furyl, 2-, 3- or 4-pyridyl and 2-, 4- or 5-thiazolyl, It is particularly preferred that RI is H.
In preferred embodiments p is 2 or 3..
It is preferred to select R2 from H, OH, CH3, CH2CH3, CH{CH3)2, CEiCH2OH)2, CH(OH)CH3 (both enantiomers) , OCH3 and OCH2CH2OH.
Moreover, it is preferred to select R3 from H, methyl, ethyl, n-propyl, i-propyl and i-amyl.
In the most preferred embodiment, said compound having the formula (I) is selected from a group consisting of (SEQ ID NOs: 1-7, respectively, in order of appear-

H-Cys-Phe-Ile-Hgn-Asn-Cys-Pro-Orn(i-Pr)-Gly-NH2
y
H-Cys-Phe-Ile-Asn { (CH2) 3OH) -Asn-Cys-Pro-Orn-Gly-NH;,
(2)

H-Cys-Phe-Ile-Asn-Asn~Cys-Pro-Dbu-Gly-NH2

H-Cys-Phe-Ile-Asn (CH2CH3) -Asn-Cys-Pro-Dbu-Gly-NH2 I _ I

(4 )

H-Cys-Phe-Ile-Gln-Asn-Cys-Pro-Orn (i-Pr) -Gly-NH2 and
H-Cys-Phe-Ile-Gln-Asn-Cys-Pro-Orn(CH2CH3) -Gly-NH2,
I I :6>
H-Cys-Phe-Iie-Asn(CH3}2-Asn-Cys-Pro-Orn-Gly-NH2
I J (7)
The number in parenthesis denotes the compound as referered to in the following.
Furthermore the present invention relates to a compound as set forth above for use as a pharmaceutical.
Accordingly, the present invention also relates to a pharmaceutical composition comprising a compound as set forth above as active ingredient in association with a pharmaceutically acceptable adjuvant, diluent or carrier.
The pharmaceutical composition may be adapted for oral, intravenous, topical, intraperitoneal, nasal, buccal, sublingual or subcutaneous administration cr for administration via the respiratory tract e.g. in the. form of an aerosol or an air-suspended fine powder. The composition may thus for instance be in the form of tablets, capsules., powders, microparticles, granules, syrups, suspensions, solutions, transdermal patches or suppositories.
It .should be noted that the composition according to the present invention may optionally include two or more of the above outlined compounds.

The present pharmaceutical composition may optionally comprise e.g. at least one further additive selected from a disintegrating agent, binder, lubricant, flavoring agent, preservative, colorant and any mixture thereof. Examples of such and other additives are found in "%Handbook of Pharmaceutical Excipients"; Ed. A.H. Kibbe, 3rd Ed., American Pharmaceutical Association, USA and Pharmaceutical Press UK, 2000.
The present pharmaceutical composition is most preferably adapted for parenteral administration. It may comprise a sterile aqueous preparation of the compounds of the invention preferably isotonic with the blood of the recipient. This aqueous preparation may be formulated according 10 kn.own methods using suitable dispersing or wetting agents and suspending agents. The injectable aqueous formulation Remestyp® (terlipressin) is exemplary of a suitable pharmaceutical formulation. The preparation may also be a sterile injectable solution or suspension in a diluent or solvent, for example as a solution in 1,3-butane diol. Water, Ringer's solution, and isotonic sodium chloride solution are exemplary acceptable diluents.. Sterile, fixed oils may be employed as a solvent or suspending medium. Bland fixed oils, including synthetic mono or di-glycerides, and fatty acids, such as oleic acid, may also be used.
In addition, the present invention relates tc use of a compound as outlined above for the manufacture of a medicament for treatment of shock of hypovolemic or vasodi-latory origin, BEV, HRS, cardiopulmonary resuscitation, anesthesia-induced hypotension, orthostatic hypotension, paracentesis-induced circulatory dysfunction, intra-operative blood loss or blood loss associated with burn debridement and epistaxis, and for treatment of various ocular diseases by increasing lacrimation/tear formation.
In another embodiment the invention relates to a method for treatment of shock of hypovolemic or vasodila-tory origin., BEV, .HRS, cardiopulmonary resuscitation, an-

esthesia-induced hypotension, orthostatic hypotension, paracentesis-induced circulatory dysfunction, intra-operative blood loss or blood loss associated with burn debridement and epistaxis, and of various ocular diseases by increasing lacrimation/tear formation, wherein said method comprises administering to an animal, including human, patient of a therapeutically effective amount of a compound as outlined above.
The typical dosage of the compounds according to the present invention varies within a wide range and will depend on various factors such as the individual needs of each patient and the route of administration. The dosage administered by infusion is generally within the range of 0.01-200 ug/kg body weight per hour. A physician of ordinary skill in the art will be able to optimise the dosage to the situation at hand.
The abbreviations used are:
Abu 2-aminobutyric acid
Boc tert-butoxycarbonyl
BOP benzotriazol-1-yloxy "trisdimethylamino-
phosphonium hexafluorophosphate
Dbu 2,4-diaminobutyric acid
DCC N,N'-dicyclohexylcarbodiimide
DCHA dicyclohexylamine
DCM dichloromethane
DIAD diisopxopyl diazodicarboxylate
DIG W/W-diisopropylcarbodiimide
DIEA N, N-diisopropyl-JV-ethylamine
DMF Nr jW-dimethylformamide
Fm 9-fluorenylmethyl
Fmoc S-fluorenylmethoxycarbonyl
Hgn homoglutamine
Hmp 2-hydroxy-3-mercaptopropionic acid
HOBt 1-hydroxybenzotriazole
HPLC high performance liquid chromatography
i iso

Mint 4-methoxytrityl
Mob p-methoxybenzyl
MS mass spectrometry
Orn ornithine
Ph phenyl
Pr propyl
PyBOP benzotriazol-1-yloxy trispyrrolidine-
phosphonium hexafluorophosphate o-NBS-Cl2-nitrobenzenesulfonyl chloride
OT oxytocin
Rt retention time
TFA trifluoroacetic acid
TIS triisopropylsilane
TMOF trimethylorthoformate
TPP triphenylphosphine
Trt trityl
VT vasotocin, [lie3] vasopressin
Unless otherwise specified L-amino acids were used, and conventional amino acid terminology is adhered to.
Experimental (synthesis)
Amino acid derivatives and resins were purchased from commercial providers (Novabiochem, Bachem Peptide International and PepTech Corporation). Fmoc-Hgn-OH was synthesised according to literature (Wisniewski, K., Kolodziejczyk, A..S, Org.. Prep. P-roced. Int.. 1997, 29, 338-341). Other chemicals and solvents were provided from Sigma-Aldrich, Fisher Scientific and VWR.
The compounds herein were synthesised by standard methods in solid phase peptide chemistry utilising both Fmoc and Boc methodology.. Unless otherwise provided, all reactions were performed at room temperature. In addition to the references cited supra, the following standard reference literature provides further guidance on general experimental set up, as well as on the availability of required starting material and reagents;:

Kates, S.A., Albericio, F., Eds., Solid Phase Synthesis. A Practical Guide, Marcel Dekker, New York, Basel, 2000;
Stewart, J.M., Young, J.D. Solid Phase Synthesis, Pierce Chemical Company, 1984;
Bisello, et al., J. Biol. Chem, 199B, 273, 22496-22505; and
Merrifield, J, Am. Chem, Soc. 1963, 85, 2149-2154.
Purity of the synthesized peptide may be determined by analytical reversed phase HPLC. Structural integrity of the peptides may be confirmed using amino acid analysis and electrospray mass spectrometry.
The peptides synthesised by Fmoc methodology were cleaved with a TFA/TIS/H20 96/2/2 (v/v/v) solution, and cleavage in Boc methodology was accomplished with 90% HF/10% aniscle (v/v) solution. Disulfide bridge (ring) formation was achieved by oxidation of linear peptides dissolved in 10% TFA (aq) with iodine. Peptides were purified by preparative H?LC in triethylammonium phosphate buffers (aq}. The compounds were finally converted to acetate salts using conventional HPLC methodology. The fractions with a purity exceeding 97% were pooled and lyophilised.
Synthesis of peptides with alkylated side chain in position no, 8:
The peptides were assembled with Fmoc methodology. The diamrno acid residue in position no, 8 was introduced with an acid labile (i.e. removable with a solution containing 1-2% TFA) protecting group, such as methoxy-trityl (Mmt; see Barlos, K. et al. in Peptides 1992, Schneider, C.H., Eberle, A.N., Eds., ESCOM Science Publishers 3-V.r 1993, pp 263-284). Resin bound peptide was treated with a DCM/TIS/TFA 93/5/2 (v/v/v) solution for the Mmt group removal. Reductive alkylation with acetone/NaBH(OAc)s provided the W-isopropyl peptide.
To avoid undesirable N,N-d±alkylation in reductive alkylation in the above procedure, which .may occur when

straight chain alkyl aldehydes are used, an alternative was developed, wherein after the Mmt removal the amino group was first derivatised with 2-nitrobenzenesulfonyl chloride (o-NBS-Cl; see Fukuyama, T.; Jow, C.-K.; Cheung, N. Tetrahedron Lett. 1995, 36, 6373-6374). The resulting sulphonamide was then alkylated with an appropriate alcohol under conventional Mitsunobu reaction conditions, typically utilising TPP/DIAD in 1,2-dimethoxyethane (Mitsunobu, 0. Synthesis 1981, 1-2-8) . The o-NBS-Cl group was subsequently removed with 5% potassium thiophenolate ±r. DMF, after which the peptide was cleaved from the resin.
Synthesis of peptides with Ar-alkylated side chair, in position no. 4:
The peptides were assembled with Boc methodology. The residue in position no. 4 was introduced in the sequence as 3oc-Asp(OFm)-OH. After complete peptide assembly the side chain protection was removed with 30% piperidine in DMF. The resulting free carboxylic group was converted to the desired amide by coupling with an appropriate amine mediated by PyBOP or BOP/DIEA. The K-terminal Boc group was then removed, followed by EF cleavage, cyclisation and purification by HPLC,
Table 1 lists the compounds prepared by the above procedure, RI is H for all compounds except no- 7, where RI is CHa- An asterisk "*" marks the most preferred embodiments.
(Table Remove) 1. Compounds prepared with the formula (I|
(Table Remove)
The following detailed examples are provided tc further illustrate the synthesis:
Compound 1; [Phe2, Hgn4,0rn (i-Pr)e] VT:
The amino acid derivatives used were Boc-Cys(Trt) -OH, Fmoc-Phe-OH, Fmoc-Ile-OH, Fmoc-Hgn-OH, Fmoc-Asn(Trt)-OK, Fraoc-CysETrt)-OHf Fmoc-Pro-OH, Fmoc-Orn(Mmt)-OH and Fmoc-Gly-OH. Fmoc-Hgn-OH was synthesised as mentioned above. Analytical HPLC was performed on a Waters 600 Liquid Chromatograph using a Vydac C18, 5u 4.6 x 250 mm, column at a flow rate of 2 ml/min. Preparative HPLC was performed on a Waters 2000 Liquid Chromatograph using a Prepak 47 x 300 mm cartridge at a flow rate of IOC ml/min, Final compound analysis was performed on a 1100 Agilent Liquid Chromatograph using a Vydac C18r 5ja 2,1 x 250 mm, column at a flow rate of 0.3 ml/min. Mass spectra were recorded on a Finnigan MAT spectrometer.
The fully protected peptide resin was synthesised on an Applied Biosystems 9050 Peptide Synthesiser starting from 2 g (0.5 mmol) of Tentagel-S-RAM resin (Peptides International). DIC/HOBt mediated single couplings with a 4-fold excess of amino acid derivatives were performed. The Fmoc group was removed with 20% piperidine in DMF. Upon completion of the automated synthesis, the resin was transferred into a manual synthesis vessel and was treated with DCM/TIS/TFA 93/5/2 (v/v/v) solution (30 ml) for 2 x 1.5 hours for removal of the Mmt group. The resin was thoroughly washed with DCM and was subsequently
suspended in 15 ml of 1,2-dichloroetehane/TMOF 1:1 (v/v) . 0.2 ml of acetone was then added followed by 0,6 g :;f NaBH(OAc)3. The suspension was shaken overnight and \:he resin was washed with methanol, DMF and DCM and dried in vacua. The resin was then treated with 30 ml of the TFA/TIS/%0 96/2/2 (v/v/v) solution for 1.5 hours and filtered off. The filtrate was evaporated and the crude linear peptide was precipitated with diethyl ether. The precipitate was immediately dissolved in 500 ml of 10% TFA (aq), and the peptide was oxidised by adding 0.1 M T2 in methanol to the magnetically stirred solution unzil yellow color persisted. Excess of iodine was reduced with ascorbic acid. The reaction mixture was then cooled with crushed ice and pH was adjusted to about 5 by'adding concentrated ammonia (aq) . The mixture was loaded onto an HPLC column and purified using a triethylammonium phosphate buffer with pH 5.2. The compound was eluted with a gradient of acetonitrile. The fractions with a purity exceeding 97% were pooled, and the resulting solution was diluted with 2 volumes of water. The solution was reloaded onto the column which was then washed with 2 1 of 0,1 M ammonium acetate (aq) and equilibrated with 2% acetic acid (aq). The compound was eluted with a fast (3%/min) gradient of acetonitrile. The fractions containing the desired product were pooled and lyophillsed.. 168 mg (-30% yield) of white amorphous powder was obtained. HPLC: Rt=8.5 min, gradient; 20-. Compound 4? [Phe2,Asn (Et)4, Dbu8]VT :
The amino acid derivatives used were Boc-Cys(Mob)-OH, Boc-Phe-OH, Boc-Ile-OH, Boc-Asp(OFm)-OH, Boc-Asn-OH, Boc-Pro-OH, Boc-Dbu(benzyloxycarbonyl)-OH DCHA salt and Boc-Gly-OH, all purchased from Novabiochem and Bachem.. HPLC and MS operations were performed as in the synthesis of compound 1.
The fully protected peptide resin was manually synthesiseti starting from 0.6 g (0.4 mmol) of 4-methyl-benzhydrylamine resin (Novabiochem). DCC, PyBOP or DIC/HOBt mediated single couplings with 2.5-fold excess of amino acid derivatives were employed. The Boc group was removed with 50% TFA in DCM containing 1% of m-cresol. Upon completion of the synthesis/ the 9-fluoren-ylmethyl ester was removed from the p-carboxylic group of aspartic acid by treatment with 30% piperidine in DKF for 2 x 30 min. The resin was washed with 1 M HOBt in DMF solution for 30 rain and then twice with DMF only. The free carbcxylic group was amidated by overnight trea-ment with 2 mmcl of ethylamine/PyBOP/DIEA in DMF. The finished resin was washed with methanol, DMF and DCM and dried in vacua. The peptide was cleaved from the resin by using 30 ml of anhydrous HF containing 3 ml of anisole at 0°C for 90 minutes. The HF was evaporated off, and the crude linear pep-ide was washed with diethyl ether. The peptide was immediately dissolved in 200 nl of 25% acetonitrile/10% TFA (aq) and oxidised as described supra. The resulting mixture was loaded directly onto an HPLC column and purified using triethylammonium phosphate buffer at pH 2.3. The subsequent purification steps were identical to the procedure for compound 1^ 41 mg (-10% yield) of white amorphous powder was obtained. HPLC: Rt=10.0 min, gradient.: 20-*40% B over 20 min, t=40°C, solvent A 0.01% TFA (aq), solvent B 70% CH3CN, 0.01% TFA (ag) ; Purity: 100%; MS (M-t-H4) : expected 992,5, observed 992.2.
The other compounds were prepared by analogous variation of these synthetic procedures. ^xperimer.tal (biological testing)
In vritro receptor assays:
Agonist activity of compounds on the hVla receptor was determined in a transcriptional reporter assay by transiently transfecting a hVla receptor expression DNA into HEK-293 cells in concert with a reporter DNA
containing intracellular calcium responsive promoter elements regulating expression of firefly luciferase. See Boss, V., Talpade, D.J., Murphy, T.J. J. Bid. Chem. 1996, May 3; 271(18), 10429-10432 for further guidance on this assay. Cells were exposed to serial dilutions of compounds diluted 10-fold per dose for 5 hours, followed by lysis of cells, determination of luciferace activity, and determination of compound efficacies and ECso values through non-linear regression. Arginine-vasopressin (AVP) was used as an internal control in each experiment, ana compounds were tested in at least three independent experiments. To determine selectivity, compounds were tested in luciferase-based tr&r.scriptional reporter assays expressing the human oxytocin (hOT) receptor. Assays for other receptors (hV2, hVlb, rat Via and rat V2) were also conducted.
For further comparative purposes, other reference compounds used were [?he2,Qrn8]OT, terlipressin and 7180.
The structure of [?he2,OrnB]OT is (SEQ ID NO: 51} :
H-Cys-?he-Ile-Gln-Asn-Cys-Pro-Orn-Gly-NH2
T
The structure of F180 is (SEQ ID NO: 52): Hmp-Phe-Ile-Bgn-Asn-Cys-Pro-Dbu(Abu)-Gly-NH2
The results of the in vitro assays are depicted in table 2 inr"ra. The EC5o value given is the geometric mean expressed in nanomol/L (nM). Selectivity values are given as ECso ratios.
In vivo pharmacological tests:
The compounds were tested in vivo for duration of action related to a standard dose of AVP. Blood pressure tests were carried out on anaesthetised Sprague-Dawley male rats (weighing 270-300 g) with catheterised jugular vein and carotid artery. The catheterised carotid artery
WES used to continuously monitor blood pressure and the jugular vein was used for administration of the compounds tested. Rats received intravenous injections of dibenair.ine prior to dosing to enhance their responsiveness to Via receptor agonists (cf. Dekanski, J., Br. J. Pharmacol. 1952, 1, 567-572). The dosing procedure consisted of one intravenous injection of physiological saline followed by two consecutive injections of a standard dose of AVP (0.1 nmol/kg, * ED7o) , and three to five increasing doses of a giver, compound selected to give at least a response comparable to the standard dose of AVP. Dosing intervals were set as time for the blood pressure to decrease to a stable baseline.
Determination of duration of action was based on the decay rate of diastolic arterial blood pressure transient increase. Specifically, for an exponential decay of plasma concentration, it can be shown that, if the response is measured beyond the distribution phase, the rate of decay near the ECso is linear and inversely proportional to the elimination half-life (Rowland, K. ani Tczer, T. in vClinical Pbzrmacokinetics, Concepts and Applications", 3rd ed., Lippincott Williams & Wilkins, Philadelphia, 1995},
To measure the response decay rate for a given compound, a dose was selected that gave an amplitude of response as similar as possible to the amplitude of response to the second injection of the standard dose of AVP. To normalise for inter-individual variation in Vla-responsiveness, the duration of action was expressed as the ratio of decay rate for this reference AVP response to the decay rate for the eguieffective dose of compound for each rat tested. The results obtained for the compounds tested are set forth in table 2.
Table 2. Results of biological testing

(Table Remove)
(Table Remove)
- = not tested
•f = selective hVla receptor agonist; ECso hOT/r.Vla ratio not determined due to very low agonist efficacy ( All references listed are to be regarded as ar. integral part of the present writ.

CLAIMS
1. A compound having the formula (I) (SEQ ID NO
53) :

(Figure Remove)
wherein:
Ar is an aryl group selected from aromatic carbocyclic
ring systems, five- or six-meinbered he^eroaromatic ring
systems and bicyclic heteroaromatic ring systems;
IR is selected from I, 2 and 3;
n is selected from 0, 1, 2, 3 and 4;
p is selected from 2, 3 and 4;
Rir R2 and RS are independently selected from H, OH,
alkyl, 0-alkyl and OC(0)-alkyl;
alkyl is selected from Ci_e straight and C4_a branched
chain alkyl and optionally has at least one hydroxyl
substituent;
and when n=0, RI and Ra optionally together form a
nitrogen containing ring structure comprising from 2 to 5
carbon atoms;
with the proviso that when Ar is phenyl, m=2, n=0,
Ri=R2=H, ana p is 3 or 4 RS is not H; and
solvates and pharmaceutically acceptable salts thereof.
2. A compound according to claim 1, wherein Ar is
selected from phenyl, 2- or 3-thienyl, 2- or 3-furyl, 2-,
3- or 4-pyridyl and 2-, 4- or 5-thiazolyl.
3, A compound according to any one of claims 1-2,
wherein RI is H.
.4- A compound according to any one of claims 1-3, wherein p is 2 or 3.

5. A compound according to any one of claims 1-4,
wherein R2 is selected from H, OH, CH3, CH2CH3, CH(CHs):f
CH{CH2OH)2, CK(OH)CH3, OCH3 and OCH2CH2OH.
6. A compound according to any one of claims 1-5,
wherein RS is selected from H, methyl, ethyl, n-propyl,
i-propyl and i-amyl.
7. A compound according to any one of claims 1-6,
wherein the compound is selected from a group consisting
of (SEQ ID NCs: 1-7, respectively, in order of appear
ance) :
H-Cys-Phe-IIe-Hgn-Asn-Cys-Pro-Orn(i-Pr)-Gly-NH2

r
H-Cys-Phe-Ile-Asn ( (CH2) 3OH) -Asn-Cys-Pro-Orn-Gly-NK2

H-Cys-Phe-Iie-Asn-Asn-Cys-Pro-Dbu-Gly-NH2
K-Cys-Phe-lle-Asn (C.H2CH3) -Asn-Cys-Pro-Dbu-Gly-NH2
H-Cys-Phe-Iie-Gln-Asn-Cys-Pro-Orn(1-Pr)-Gly-NH2 and
H-Cys-Phe-Ile-Gln-Asn-Cys-Pro-Orn (CH2CH3) -Gly-NH2.
8. A compound according to claim 1 having the structural formula:
H-Cys~Phe-Ile-Asn(CH3)2-Asn-Cvs-Pro-Orn-Gly-NH2
9. A compound according to any one of claims 1-8 for
use as a pharmaceutical-
10. A pharmaceutical composition comprising a
compound according to any one of claims 1-8 as active
ingredient in association with a pharmaceutically
acceptable adjuvant, diluent or carrier..

11. Use of a compound according to any one of claims
1-8 for the manufacture of a medicament for treatment of
shock of hypovolemic or vasodilatory origin, bleeding
esophageal varices, hepatorenal syndrome, cardiopulmonary
resuscitation, anesthesia-induced hypotension, ortho-
static hypotension, paracentesis-induced circulatory dys
function, intra-operative blood loss or blood loss asso
ciated with burr, debridement and epistaxis, and for
treatment of various ocular diseases by increasing lacri-
mstion/tear formation.
12. A method for treatment of shock of hypovolerr.ic
or vasodilatory origin, bleeding esophageal varices,
hepatorenal syndrome, cardiopulmonary resuscitation, an
esthesia-induced hypotension, orthostatic hypotension,
paracentesis-induced circulatory dysfunction, intra-
operative blood loss or blood loss associated with burr.
debridement and epistaxis, and of various ocular diseases
by increasing lacrimation/tear formation, wherein said
method comprises administering to an animal, including
human, patient of a therapeutically effective amount of a
compound according to any one of claims 1-8.

cant

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Patent Number

258244

Indian Patent Application Number

6096/DELNP/2006

PG Journal Number

52/2013

Publication Date

27-Dec-2013

Grant Date

20-Dec-2013

Date of Filing

18-Oct-2006

Name of Patentee

FERRING B.V

Applicant Address

POLARIS AVENUE 144, NL-2132 JX HOOFDDORP NETHERLANDS

Inventors:

#	Inventor's Name	Inventor's Address
1	WISNIEWSKI KAZIMIERZ	12793 VIA TERCETO, SAN DIEGO, CA 92130 USA.
2	SCHTEINGART CLAUDIO	6912 FISK AVENUE, SAN DIEGO, CA 92122 USA.
3	LAPORTE REGENT	11344 TRILLIUM WAY, SAN DIEGO, CA 92131 USA.
4	GALYEAN ROBERT FELIX	420 ROCK RIDGE PLACE, ESCONDIDO, CA 92027 USA.
5	RIVIERE PIERRE	3993 VIA CANGREJO, SAN DIEGO, CA 92130 USA.

PCT International Classification Number

A61K 38/00

PCT International Application Number

PCT/US2005/027772

PCT International Filing date

2005-08-03

PCT Conventions:

#	PCT Application Number	Date of Convention	Priority Country
1	60/600,377	2004-08-11	EUROPEAN UNION
2	04019029.0	2004-08-11	EUROPEAN UNION