CN1901941A - Conjugates of angiotensin ii and an imaging moiety - Google Patents

Abstract

The invention comprises pharmaceuticals of formula (I) Z-(L)n-V, wherein V denotes a peptide, L denotes an optional linker, Z denotes a group that optionally can carry an imaging moiety M, n denotes 0 or 1. The pharmaceuticals are active as therapeutic agents for the treatment of heart failure, cardiac arrhythmias and diseases of fibrosis prominent such as COPD, liver fibrosis and atherosclerosis. The novel pharmaceutical comprises targeting moieties bonded to the receptor which receives up regulation and/or excess expression in or not in siclen region. The imaging moieties contained in the targeting moieties carries imaging moieties which can form image for diagnosis, optional linkers and peptides. The novel pharmaceutical compound has high affinity to angiotensin receptor, in particular to angiotensin II(AngII)I type(AT1)receptor.

Description

Conjugates of angiotensin II and imaging moieties

field of invention

The present invention provides novel drugs for effective treatment of heart failure, arrhythmia and other fibrosis-prominent diseases, as well as for diagnosis of fibrosis-prominent diseases and symptoms. The invention also provides novel pharmaceutical compositions and precursors for preparing diagnostic agents. In addition, the present invention also provides novel drugs for effective monitoring of therapy and methods of monitoring therapy. Still further objects of the invention are apparent from the claims.

The novel drugs comprise targeting moieties that bind to receptors that may or may not be upregulated and/or overexpressed in the disease area. The targeting moiety comprises an imaging agent moiety with a diagnostically imageable moiety, an optional linker group and a peptide moiety.

The novel pharmaceutical compound has high affinity for angiotensin receptors (Angiotensin Receptors), especially for angiotensin II (Ang II) type 1 (AT ₁ ) receptors.

Background of the invention

Angiotensin II (Ang II)-octapeptide (Asp-Arg-Val-Tyr-Ile-His-Pro-Phe)-is a pleiotropic vasoactive peptide that interacts with two different receptors: Ang II Type 1 (AT ₁ ) and type 2 (AT ₂ ) receptors bind. Activation of the renin-angiotensin-aldosterone system (RAAS) leads to vascular hypertrophy, vasoconstriction, salt and water retention, and hypertension. These effects are mainly mediated by the AT ₁ receptor. Paradoxically, other Ang II-mediated effects, including cell death, vasodilation, and natriuresis, are mediated by _AT2 receptor activation. The mechanism of Ang II signaling is still not fully understood. _AT1 receptor activation triggers a variety of intracellular systems, including tyrosine kinase-induced protein phosphorylation, production of arachidonic acid metabolites, changes in the activity of reactive oxidant species, and fluctuations in intracellular Ca ⁺ concentrations. Activation of AT ₂ receptors results in excitation of bradykinin, nitroxide production, and prostaglandin metabolism, which are in large part opposite to those of AT ₁ receptors. (See: Berry C. Touyz R, Dominiczak AF, Webb RC, Johns DG.: Am J Heart Circ Physio. 2001 Dec; 281(6): H2337-65. Angiotensin receptors: signaling, vascular pathophysiology, and neuronal amide interactions).

Ang II is the active component of the renin-angiotensin-aldosterone system (RAAS). It has important physiological roles in the regulation of blood pressure, plasma volume, sympathetic nerve activity, and thirst response. Ang II also has a pathophysiological role in cardiac hypertrophy, myocardial infarction, hypertension, chronic obstructive pulmonary disease, liver fibrosis, and atherosclerosis. It produces systemic effects through the classical RAAS and local effects through the tissue RAAS. In classical RAAS, circulating nephrogenic renin cleaves hepatic angiotensinogen to form the decapeptide angiotensin I (Ang I), which can be converted in the lung to Active Ang II. Ang I can also be processed into the heptapeptide Ang-(1-7) by tissue endopeptidases.

The RAAS system is shown schematically herein in Figure 1, which is based on Figure 1 in Foote et al., Ann. Pharmacother. 27 :1495-1503 (1993).

In addition to the important role of RAAS in normal cardiovascular homeostasis, the excessive activity of RAAS is also related to the development of various cardiovascular diseases such as hypertension, congestive heart failure, coronary ischemia and renal insufficiency. After myocardial infarction (MI), the RAAS is activated. In particular the AT ₁ receptor appears to play a major role in post-MI remodeling, as AT ₁ receptor expression is increased after MI and in left ventricular dysfunction. Therefore, those drugs that interfere with the RAAS, such as ACE inhibitors and _AT1 receptor antagonists, have been shown to have powerful therapeutic effects in such cardiovascular diseases.

For heart, kidney, lung and liver etc., fibrosis is a common factor in the failure of said organs. There is therefore considerable interest in understanding the pathophysiological mechanisms involved in organ fibrosis, especially in terms of enabling protective pharmacological strategies. Tissue repair involves inflammatory cells, including members of the monocyte/macrophage lineage, integrally initiating the repair process; and myofibroblasts, phenotype-switching mesenchymal fibroblasts, responsible for collagen renewal and fibrous tissue formation. Each of these cells present in the repair microenvironment is associated with molecular events leading to the de novo generation of angiotensinogen II (Ang II). In an autocrine/paracrine manner, these peptides modulate the expression of TGF-β1 through angiotensinogen (AT ₁ ) receptor ligand binding. It is this cytokine that changes the phenotype of fibroblasts to myofibroblasts (myoFb) and regulates the renewal of collagen by myofibroblasts. Angiotensin-converting enzyme (ACE) inhibition or AT ₁ receptor antagonism, respectively, prevent many of these fibrogenic molecular and cellular responses and have thus been considered protective interventions. (See: Weber KT. Fibrosis, a common to organ failure: angiotensin II and tissue repair. Semin Nephrol. 1997 Sep; 17(5): 467-91 and references therein).

Ang II can regulate tissue fibrosis by activating mesenchymal cells. For example, Ang II stimulates the proliferation of cardiac fibroblasts in vitro by activating _AT1 . The presence of _AT1 receptors has been shown on cardiac fibroblasts in vitro. Most of the profibrotic actions of Ang II appear to be mediated through this receptor; however, increased expression of AT ₂ on cardiac fibroblasts has been detected in hypertrophic human hearts, and there is a correlation between the expression of these two isoforms. Balance may be critical for determining Ang II responses. (See: Am.J.Respir.Crit.Care Med., Volume June 2000, 1999-2004Angiotensin II is Mitogenic for HumanLung Fibroblasts via Activation of the Type 1 Receptor RICHARD P.MARSHALL, ROBIN J.MCANULTY, and GEOFFREY J.LAURENTand references therein).

The Ang II receptors can be differentiated on the basis of inhibition by specific antagonists. AT ₁ receptors can be selectively antagonized by biphenylimidazoles, such as Losartan, while tetrahydroimidazopyridines specifically inhibit AT ₂ receptors. The _AT2 receptor was also selectively activated by CGP-42112A (which is a hexapeptide analog of AngII, which also inhibits the _AT2 receptor, depending on the concentration). Two other angiotensin receptors have been described: the AT ₃ and AT ₄ subtypes.

In rodents, the _AT1 receptor has two functionally distinct subtypes, _AT1A and _AT1B , with >95% amino acid sequence identity.

The second major angiotensin isoform is the AT ₂ receptor. It has low amino acid sequence homology (-34%) to the AT _1A or AT _1B receptors. Although the precise signaling pathways and functional roles of AT ₂ receptors remain unclear, these receptors may antagonize AT ₁ -mediated effects under physiological conditions by inhibiting cell growth and by reducing apoptosis and vasodilation. The precise role of the AT ₂ receptor in cardiovascular disease remains to be elucidated.

In addition to AT ₁ and AT ₂ , other Ang II receptors are known and are commonly referred to as AT _atypical (see Kang et al., Am. Heart J. 127 :1388-1401 (1994)).

Inhibition of the action of Ang II has been used medically, for example in the management of hypertension and heart failure. This has been achieved in several ways: by using renin inhibitors that block the conversion of angiotensinogen to angiotensin I (the precursor of Ang II); by using renin inhibitors that block the conversion of angiotensin I to Ang II (and Angiotensinase (ACE) inhibitors that also block the biotransformation of bradykinin and prostaglandins); by using anti-Ang II-antibodies; and by using Ang II antagonists.

Beta-blockers are most commonly used in antiarrhythmic therapy. Antiarrhythmic drugs have limited overall success and calcium channel blockers can sometimes cause arrhythmias. No single drug has shown superiority, with the possible exception of amiodarone. Short-term antiarrhythmic benefits, depending on the specific drug, have been found to be outweighed by their neutral or negative effect on mortality. (Sanguinetti MC and Bennett, PB: Target selection and screening of antiarrhythmic drugs. Circulation 2003, 93(6): 491-9257-263). There is clearly a need for better antiarrhythmic drugs.

The Lancet publication (Llindholm, LH et al. Effect of Losartan on sudden cardiac death in diabetic patients: data from the LIFE study. The Lancet, 2003, 362:619-620) reveals the AT ₁ receptor Antagonists, besides being generally beneficial for patients with CHF, also reduce the incidence of sudden cardiac death. Some studies have shown that AT ₁ receptor antagonists have antiarrhythmic effects on the arrhythmic effects induced by myocardial infarction or reperfusion after LAD ligation (Harada K et al. Angiotensin IIa receptor type and reperfusion rhythm Circulation.1998, 97:315-317. Ozer MK et al. Captopril and rosartan in arrhythmia and gangrene caused by myocardial ischemia-reperfusion in mice. Pharmacological research, 2002 , 45(4), 257-263 Lynch JJ et al., EXP3174, a fully antagonistic human metabolite of rosartan, but neither rosartan nor the angiotensin-converting enzyme inhibitor captopril, avoids in dogs with recent myocardial infarction Lethal ischemic arrhythmias in a model. JACC, 1999, 34876-884).

Description of related fields

WO 98/18496 (Nycomed Imaging AS) discloses contrast agents comprising a vector linker-reporter configuration, wherein the vector comprises angiotensin or a peptide angiotensin derivative.

US Patent 4411881 (New England Nuclear Corporation) describes the stabilization of radio-labeled compounds. Examples of radioactively-labeled compounds include, for example, angiotensin II (5-L-isoleucine) [tyrosyl- ¹²⁵ I]-(monoiodide).

Ang II can be converted into potent antagonists or partial antagonists by altering their amino acid composition. For example, substituting isoleucine for phenylalanine at position 8 and sarcosine for aspartic acid at position 1 turned the peptide into a potent antagonist.

Affinity-specificity to the AT ₁ receptor can be increased by cyclizing or bridging amino acids at positions 4 and 6. Similarly, introduction of sarcosine at position 1 and glycine at position 8 converts the peptide into an AT ₁ selective antagonist (see RC Speth. Sarcosinell, Glycine 8 Angiotensin II is the AT ₁ Angiotensin II Receptor Subtype-selective antagonists Regulatory peptides 115 (2003) 203-209).

As mentioned above, the natural ligand of the _AT1 receptor is the octapeptide Ang II, Asp-Arg-Val-Tyr-Ile-His-Pro-Phe, which binds to the _AT1 receptor in the nanomolar range.

The avidity of the peptide carrier is often compromised when a natural binding ligand is modified by combining a moiety with other moieties, especially relatively bulky and bulky moieties.

We surprisingly found that the octapeptide AngII, Asp-Arg-Val-Tyr-Ile-His-Pro-Phe and its derivatives, when substituted at specific positions of the peptide, not only retained its binding ability, but also Surprisingly increased affinity for angiotensin II receptors, especially for AT ₁ receptors.

Summary of the invention

The first object of the present invention is to provide a drug for effective treatment of heart failure, arrhythmia and other fibrosis-prominent diseases, such as COPD, liver fibrosis and arteriosclerosis, which drug comprises a targeting moiety, which has been shown to inhibit AT ₁ receptor has a stronger affinity than the natural octapeptide Ang II. The drug should exhibit antagonistic activity.

A second object of the present invention is to provide medicaments effective in the diagnosis of heart failure and other fibrosis-prominent diseases, such as in COPD, liver fibrosis and arteriosclerosis, said medicaments comprising a targeting moiety that binds an imageable moiety. The imageable moiety may be any imageable moiety which, when administered to a subject, produces an image of at least a portion of the subject where the contrast agent is distributed, e.g. by radiographic imaging, SPECT , PET, MRI, X-ray, Optical Imaging (OI), Ultrasound (US), Electron Impedance or Magnetic Imaging modalities. A targeting moiety combined with an imageable form should show a higher affinity for the _AT1 receptor than Ang II and should preferably be used as an antagonist, although weak agonistic activity may still be acceptable.

When the novel drug has a suitable imageable moiety for diagnostic imaging, it can be used sequentially or concurrently as a therapeutic agent.

Further objects include providing methods of treating hypertension, fibrosis, COPD and related diseases, methods of imaging heart failure and fibrosis and methods of monitoring the progress of treatment of such diseases and disorders and related cardiovascular diseases and disorders. The present invention also provides novel pharmaceutical compositions and precursors for the preparation of diagnostic agents. Also provided are diagnostic kits, particularly kits for the preparation of radiopharmaceutical diagnostics.

The medicine of the present invention comprises peptide V, optional linker L and Z part, as shown in formula (I)

Z-(L) _n -V (I)

in,

V represents a peptide with the binding sequence -X ¹ -X ² -Val-Tyr-Ile-His-Pro-X ³ ,

L indicates an optional linker,

Z represents a group which may optionally carry an imaging moiety M,

n is 0 or 1,

X ¹ represents an amino acid,

^X represents Arg or N-alkylated Arg, or a mimetic of Arg,

X ³ represents an amino acid comprising a hydrophobic side chain,

wherein each of the valine (Val) and isoleucine (Ile) residues at positions 3 and 5 can optionally be replaced by an amino acid capable of forming a bridge,

Z is optionally bonded to amino acid ^X1 through a linker L and M, when present, represents an imageable moiety which can be detected directly or indirectly in a diagnostic imaging procedure.

Detailed description of the invention

The present invention is described in the invention claims. Specific features of the invention are set forth in the following detailed description and examples.

In the targeting moiety of formula (I), the aforementioned V represents the peptide sequence -X ¹ -X ² -Val-Tyr-Ile-His-Pro-X ³ .

Unless otherwise stated, the amino acids in peptide V of formula (I) are the L-amino acids in native Ang II.

The three letter abbreviations used for amino acids have the following meanings:

Arg - Arginine

Asp - Aspartic Acid

Cys - cysteine

Hcy - homocysteine

Gly - Glycine

Sar - Sarcosine

Val - Valine

Tyr - Tyrosine

Ile - Isoleucine

His - Histidine

Pro - Proline

Phe - Phenylalanine

Abu -2-amino-butyric acid

Nva -2-amino-valeric acid

Nle -2-amino-caproic acid

Phg -2-amino-2-phenylacetic acid

Hph -2-amino-4-phenylbutyric acid

Bip -2-amino-3-biphenylpropionic acid

Nal -2-amino-3-naphthoic acid

Cha -2-amino-3-cyclohexylpropionic acid

The amino acids of V are preferably independently selected

X ¹ represents -NY ₁ -(CH ₂ ) _m -CO-, wherein m is an integer of 1-10 and Y ₁ is H or an alkyl or aryl group containing substituents, most preferably Gly

X ² represents Arg or N-methyl-Arg or Arg mimics Phe[4-guanidino] and Gly-4-piperidinyl[N-amidino],

^X3 represents Phe, D-Phe, Ile, Abu, Nva, Nle, Phg; Hph, Bip, Nal or Cha, most preferably D-Phe, Bip, Ile or Hph.

Drugs in which ^X1 represents Gly, ^X2 represents Arg or N-methyl-Arg and ^X3 represents D-Phe, Bip, He or Hph are preferred.

Still further preference is given to medicaments wherein X ¹ represents Gly, X ² represents Arg and X ³ represents He.

If the amino acids at positions 3 and 5 are selected to form a bridge element, the bridge preferably contains _-CH2 - _CH2- , -S- _CH2- , -S- _CH2 -S-, lactam or -SS- unit. More preferably, the covalent bond is a disulfide bond formed by paired oxidation of two cysteines or homocysteines.

Examples of suitable linkers L are described in patents WO 98/18496 and WO 01/77145 (pages 23-27), the contents of which are incorporated herein by reference. L may preferably represent polyalkylene glycol units, such as polyethylene glycol (PEG) and polypropylene glycol (PPG), carbohydrates, dextran or 1-10 amino acids. The linkers may also act as biomodifying agents, as described in WO03/006491.

The linker L can also be derived from an alkylamine or an arylamine, preferably a compound of the formula NH-(CH ₂ ) _m -, optionally combined with -CO-(CH ₂ ) _m -CO-, where m Represents a positive integer from 1-10.

The linker may also comprise one or more PEG units as defined in Formula IV below, wherein n is an integer of 1-10.

Most preferred are linkers defined by formula (V) and formula (VI):

Formula (V)

Formula (VI)

The Z moiety comprises a non-peptide moiety having a molecular weight in excess of 50D (Dalton), more preferably between 100-1000D and even more preferably between 300-700D. The Z moiety can be any pharmaceutically acceptable chemical entity so long as the final targeting moiety of formula (I) exhibits a higher affinity for the AT ₁ receptor than the natural ligand Ang II. More specifically, Z represents an organic group with a suitable functional group so that Z can react with linker L or directly with peptide V to form a stable covalent bond.

Z may represent a straight-chain or branched hydrocarbon radical optionally containing one or more double or triple bonds and optionally substituted by halogen, oxygen, sulfur or phosphorus atoms or optionally comprising heterogeneous compounds such as oxygen, nitrogen or sulfur atom. More specifically, hydrocarbyl may represent substituted or unsubstituted alkyl, alkenyl, alkynyl groups having a molecular weight of at least 50D.

Z may also represent one or more linked carbocyclic residues comprising a monocyclic, bicyclic or tricyclic ring system which may be saturated, partially unsaturated or aromatic, which may be substituted or unsubstituted and Its molecular weight is at least 50D. Examples of such ring systems are aryl, aralkyl, cyclohexyl, adamantyl and naphthyl.

Z may further represent one or more linked heterocyclic compounds, such as 5, 6, 7, 8, 9 or 10-membered ring systems, which may be monocyclic, bicyclic or tricyclic and may contain one or more N, O, S and P are used as heteroatoms. Such ring systems may also be attached to hydrocarbyl and carbocyclic groups and as defined above or fused to carbocyclic groups. Examples of such groups are acridinyl, benzofuryl, indolyl, pyridyl, piperidinyl, morphoridinyl and thienyl.

Z can also represent polyalkylene glycols, such as polyethylene glycol (PEG) and polypropylene glycol (PPG), all carbohydrates with a molecular weight exceeding 50D such as mono- or polysaccharides. Polyalkylene glycols can additionally be used as biomodifying agents.

A specific Z represents a chelating agent such as acyclic or cyclic polyaminocarboxylates (e.g. DTPA, DTPA-BMA) as described in U.S. Patent 4,647,447 (Schering AG) and WO 86/02841 (Nycomed Saluta, Inc.). , DOTA and DO3A), which are hereby incorporated by reference. Chelating agents also include aminethiols such as diaminedithiols, amineoximes and hydrazines and related reagents described in patent WO 01/77145 (see Table 1 therein), incorporated herein by reference In this article. Chelating agent cPN216 of formula (VIII) is particularly preferred.

For therapeutically effective drugs, Z can be any entity as described above.

For a drug to be effective in diagnostics, especially in vivo, the Z moiety must be capable of carrying an imageable moiety or a moiety represented by M. With refers to any form of linkage between the Z and M moieties, such as chemical bonds, such as covalent or electrovalent bonds or ionic bonds or by adsorption or any other type of linkage.

Z can be any imageable portion. Wherein M refers to a metal entity and Y ₁ represents a chelating agent. The nature of Z and/or _Y1M will depend on the imaging modality used in the diagnosis. Z and/or Y ₁ M must be detectable directly or indirectly during in vivo diagnostic imaging, e.g., moieties that emit or can cause the emission of detectable radiation (e.g., by radioactive decay, fluorescence excitation, spin resonance excitation, etc.), affecting local Parts of the electromagnetic field (such as paramagnetic, superparamagnetic, ferrimagnetic, or ferromagnetic types), parts that absorb or scatter radiated energy (chromophores, particles (including vesicles containing gases or liquids), heavy elements And its compounds, etc.), and the part that produces detectable substances (such as gas microbubble generator).

In a preferred embodiment, one Z moiety is directly covalently bonded to ^X to form an N-alkylglycine unit.

Chelating agents of formula (VII) and formula (VIII) hereinafter are also particularly preferred.

A wide range of suitable imageable moieties is known from WO 98/18496, the contents of which are incorporated herein by reference.

The imaging modality and imaging sections Z and M are described in more detail below:

In a first embodiment, the compound of formula (I) comprises a _Y1 moiety with one or more imageable moieties M effective in Radio and SPECT imaging modalities. Preferred M are gamma emitters with low or no alpha- and beta-emission and have a half-life of more than one hour. Preferred M groups are radionuclear ⁶⁷ Ga, ¹¹¹ In, ¹²³ I, ¹²⁵ I, ¹³¹ I, ^81m Kr, ⁹⁹ Mo, ^99m Tc, ²⁰¹ Tl and ¹³³ Xe. Most preferred ^is99mTc .

M can be further represented by the following isotopes or pairs of isotopes for imaging and therapy without the need to alter radiolabeling methodology or chelators: ⁴⁷ Sc ₂₁ ; ¹⁴¹ Ce ₅₈ ; ¹⁸⁸ Re ₇₅ , ¹⁷⁷ Lu ₇₁ ; ¹⁹⁹ Au ₇₉ ; ⁴⁷ Sc ₂₁ ; ¹³¹ I ₅₃ ; ⁶⁷ Cu ₂₉ ; ¹³¹ I ₅₃ and ¹²³ I ₅₃ ; ¹⁸⁸ Re ₇₅ and ^99m Tc ₄₃ ; ⁹⁰ Y ₃₉ and ⁸⁷ Y ₃₉ ; ⁴⁷ Sc ₂₁ and ⁴⁴ Sc ₂₁ ; ⁹⁰ Y ₃₉ and ¹²³ I ₅₃ ; ¹⁴⁶ Sm ₆₂ and ¹⁵³ Sm ₆₂ ; and ⁹⁰ Y ₃₉ and ¹¹¹ In ₄₉ .

When M represents a metal radionuclide, then _Y represents a chelating agent suitable for forming a stable chelate with M. Such chelating agents are well known in the art and typical examples of such chelating agents are described in Table 1 of WO 01/77145.

Chelating agents of formula (VII) are particularly preferred:

in:

Each of R ¹ , R ² , R ³ and R ⁴ is independently H or C _1-10 alkyl, C _3-10 alkylaryl, C _2-10 alkoxyalkyl, C _1-10 hydroxyalkyl , C _1-10 alkylamine, C _1-10 fluoroalkyl, or 2 or more R groups, together with the atoms connecting them, form a saturated or unsaturated carbocyclic or heterocyclic ring.

A more particularly preferred chelating agent is of formula (VII), wherein R ¹ , R ² and R ³ are hydrogen or methyl and R ⁴ is an alkylamine group, most particularly a compound of formula (VIII), here represented by cPN216 .

Formula (VIII)

Most preferred for _Y1 is when the chelator is cPN216 and the imaging factor M ^is99mTc .

The synthesis of chelating agents of formula (VII) and (VIII) is described in WO 03/006070.

Other preferred chelating agents are compounds of formula (XI)

Formula (XI)

Wherein Q ₁ -Q ₆ are independent Q groups, wherein Q is H, alkyl, aryl or amine protecting group.

W ₁ is -NR-, -CO ₂ -, -CO-, -NR(C=S)-, -NR(C=O)-, -CONR- or Q group;

each Y is independently a D- or L-amino acid, -CH ₂ -, -CH ₂ OCH ₂ - or -OCH ₂ CH ₂ O-, or an X group;

p is an integer whose value is 1-8;

q is an integer whose value is 0-30;

R is H, C _1-4 alkyl, C _2-4 alkoxyalkyl, C _1-4 hydroxyalkyl or C _1-4 fluoroalkyl;

Q is

A is a counter ion;

The synthesis of the tetraamine chelating agent of formula (XI) can be found in the British patent that application number is GB 0416062.8.

Non-metallic radionuclides such as ¹²³ I, ¹²⁵ I and ¹³¹ I can be covalently attached to the L moiety (when L is present) or to X ₁ by substitution or addition reactions well known in the art.

In a second embodiment, the compound of formula (I) comprises a Z moiety that is effective in a PET imaging format. Z then denotes a radioactive emitter with positron-emitting properties. Preferred Z groups are the radionuclides ¹¹ C, ¹⁸ F, ⁶⁸ Ga, ¹³ N, ¹⁵ O and ⁸² Rb. ¹⁸ F is particularly preferred. Chelation of metal emitters ⁸² Rb and ⁶⁸ Ga with chelating agent Y ₁ is also preferred.

Thiol coupling chemistry, ¹⁸ F-synthetic elements and labeled peptides were prepared using thiol coupling chemistry as described in WO 03/080544, the contents of which are incorporated herein by reference.

A description of the labeling of peptides by employing thiol coupling chemistry can be found in British Patent Application No. 0317815.9, the contents of which are hereby incorporated by reference.

When M represents a metal radionuclide, then _Y represents a chelating agent suitable for forming a stable chelate with M. Such chelating agents are well known at this stage in the art and typical examples of such chelating agents are described in Table 1 of WO 01/77145 and above in the Radio and SPECT imaging sections.

In another preferred embodiment, Y _is a DOPT chelator and M is ^Ga , which can be easily incorporated into the chelate using microwave chemistry.

Non-metallic radionuclides such ^as18F , when present, can be linked covalently to the L moiety, or to _X1 by substitution or addition reactions known at this stage in the art, which are also described in WO03/080544 , which is hereby incorporated by reference.

In a third embodiment, the compound of formula (I) comprises a _Y1 moiety with one or more M imageable moieties effective in an MR imaging modality. Here M represents a paramagnetic metal, as described in US Patent 4647447, Gd ³⁺ , Dy ³⁺ , Fe ³⁺ and Mn ²⁺ are particularly preferred and Y ₁ represents a chelating agent, especially as described in US Patent 4 647 447 and the acyclic or cyclic polyaminocarboxylates (eg DTPA, DTPA-BMA, DOTA and DO3A) described in WO 86/02841. M can also represent metal oxides absorbed by Z, such as superparamagnetic, ferrimagnetic or ferromagnetic species, so Z is used as a coating for metal oxides. Metal oxides for use as MR contrast agents are described in US Patent 6 223 777, incorporated herein by reference.

In a fourth embodiment, the compound of formula (I) comprises a _Y1 moiety with one or more M moieties imageable in an X-ray imaging modality. M here denotes a heavy metal, such as W, Au and Bi, preferably in oxide form which can be absorbed by Z. Z can also be represented by iodinated aryl derivatives known in particular as X-ray contrast agents, such as Iopamiron ^™ and Omnipaque ^™ . These reagents can be linked to peptide V of formula (I) via their amide or amine functional groups.

In another embodiment, the compound of formula (I) comprises Z in the form of a gas-filled vesicle. Such ultrasound imaging agents may be used in receptor imaging, as when they are used in conjunction with peptides, as is known in the art, as described in WO98/18500.

In a sixth embodiment of the present invention, the Z moiety of formula (I) may be any moiety that can be detected directly or indirectly in an optical imaging procedure. The detectable moiety can be a light scatterer (eg, a colored or colorless particle), a light absorber, or a light emitter. More preferably Z is represented by a dye, such as a chromophore or a fluorescent compound. The Z moiety can be any dye that is capable of interacting with light at wavelengths from ultraviolet to near infrared in the electromagnetic spectrum. In a preferred version, Z has fluorescent properties.

Preferred organic dye moieties include groups with extensively delocalized electron systems, such as cyanines, merocyanines, indocyanines, phthalocyanines, naphthalocyanines , triphenylmethines, porphyrins, pyrilium dyes, thiapyrilium dyes, squarylium dyes, croconium dyes, azulenium dyes, indoanilines ), benzophenoxazinium dyes, benzothiaphenothiazinium dyes, anthraquinones, napthoquinones, indathrenes, ortho Phthaloylacridones, trisphenoquinones, azo dyes, intramolecular and intermolecular charge transfer dyes and dye complexes, tropones, tetrazines , bis (dithiolene (dithiolene)) complex, joint (benzene-dithiolate (dithiolate)) complex, iodoaniline dyes, bis (S, O-dithiolene (dithiolene) ))Complex. Fluorescent proteins such as green fluorescent protein (GFP) and GFP modifications with different absorption/emission properties are also effective. Complexes of certain rare earth metals (such as europium, samarium, terbium or dysprosium) are used in some cases, such as fluorescent nanocrystals (quantum dots).

A further description of suitable parts in an optical imaging procedure can be found in Norwegian Patent Application No. 200303115, the contents of which are incorporated herein by reference.

Drugs of formula (I) may be further modified by attaching one or more biomodifier groups, such as polyalkylene glycol units, such as polyethylene glycol (PEG) and polypropylene glycol (PPG). Examples of biomodifier groups are described in WO 03/006491, the contents of which are incorporated herein by reference. The biomodifier group can be connected to any position in the compound of formula (I), as long as it has no obvious negative effect on the connection of the compound to the targeting receptor.

The biomodifier is preferably based on a monodisperse PEG building block, comprising 1-10 units of the building block, and has the function of changing the pharmacokinetics and plasma clearance of the drug. In a preferred embodiment of the present invention, the compound of formula IV represents a biomodifier unit composed of monodisperse PEG-like structure polymerization, 17-amino-5-oxo-6-aza-3,9,12 , 15-tetraoxaheptadecanoic acid.

wherein n is equal to an integer of 1-10, and wherein the C-terminal unit forms an amide bond.

Examples of drugs of formula (I) are represented by the following:

Compounds of formula (II) and formula (III):

Z-CO-NH-(CH ₂ ) _m -X ¹ -X ² -Val-Tyr-Ile-His-Pro-X ³ (II)

Z-CO-(CH ₂ ) _m -CO-NH-(CH ₂ ) _m -X ₁ -X ₂ -Val-Tyr-Ile-His-Pro-X ₃ (III)

wherein m is an integer between 1-5, and Z, X ¹ , X ² and X ³ are as defined above.

Compounds defined by the formulas Va and Via:

wherein Z and V are as defined above.

Compound of formula (IX):

wherein ^Y2 is an alkyl group, an aryl group or a moiety comprising a short PEG and ^Y2 is preferably _-CH2 - _CH2 - _CH2- and ^X3 is as defined above.

Preferred examples of drugs of formula (I) are represented by:

cPN216-Gly-Arg-Val-Tyr-Ile-His-Pro-Ile-OH

cPN216-Gly-MeArg-Val-Tyr-Ile-His-Pro-D-Phe-OH

cPN216-Gly-Arg-Val-Tyr-Ile-His-Pro-Bip-OH

N-((CH ₂ ) ₆ -tetraamine)-Gly-Arg-Val-Tyr-Ile-His-Pro-Ile-OH

As noted above, chelates of compounds ^{bearing99mTc or18F} are particularly preferred for use in in vivo diagnostics.

Especially preferred is the compound cPN216-Gly-Arg-Val-Tyr-Ile-His-Pro-Ile-OH, whose chemical structural formula is represented by formula (X):

Formula (X)

And its chelate with ^99m Tc.

Particularly preferred compound N-((CH ₂ ) ₆ -tetraamine)-Gly-Arg-Val-Tyr-Ile-His-Pro-Ile-OH, its structural formula is shown by formula (XII):

Formula (XII)

And its chelate with ^99m Tc.

The medicament of formula (I) is preferably administered as a pharmaceutical formulation comprising a compound of formula (I) in a form suitable for administration to a mammal, such as a human. The administration is via formulations suitable for injection or infusion, such as aqueous solutions. The formulation may contain one or more pharmaceutically acceptable additives and/or excipients, such as buffers; solubilizers such as cyclodextrins; or surfactants such as Pluronic, Tween ) or phospholipids. In addition, stabilizers or antioxidants such as ascorbic acid, gentisic acid or p-aminobenzoic acid and bulking agents such as sodium chloride or mannitol for lyophilization may be added.

In one aspect of the invention, the medicament of formula (I) is effective in therapy and therapy monitoring. In the monitoring of heart failure and other fibrosis-prominent diseases, especially COPD, liver fibrosis and arteriosclerosis treatment progress, one of the methods is to give the treated experimenter a drug of formula (I) and subsequently treat An image is generated of the subject's whole body or a portion of the subject.

In yet a further aspect there is provided a kit for preparing the radiopharmaceutical composition of formula (I), the kit comprising a peptide-chelate conjugate and a reducing agent. A preferred reducing agent of the kit is a stannous salt. The kit may also contain one or more stabilizers, antioxidants, and bulking and solubilizing agents for lyophilization.

General Procedures for Preparation of Drugs and Their Precursors

The abbreviations used have the following meanings:

Fmoc: 9-fluorenylmethoxycarbonyl

Boc: tert-butoxycarbonyl

tBu: tert-butyl

Trt: Trityl

Pmc: 2,2,5,7,8-pentamethylchroman (chroman)-6-sulfonyl

TFA: Trifluoroacetic acid

HBTU: O-benzotriazol-1-yl-N,N,N',N'-tetramethyluronium hexafluorophosphate

DMF: Dimethylformamide

NMP: N-Methylpyrrolidone

TIS: Triisopropylsilane

NHS: N-Hydroxysuccinimide

NMM: N-methylmorpholine

RP-HPLC: Reversed Phase High Performance Liquid Chromatography

Wang resin: p-Benzyloxybenzyl alcohol resin

Synthesis of V:

The peptide V described in the present invention can be synthesized by employing all known chemical synthesis methods but particularly effective is the Merrifield solid-phase method using an automatic peptide synthesizer (J. Am. Chem. Soc., 85: 2149 (1964) ). Typically, the desired sequence is assembled by solid phase peptide synthesis. Standard procedures for the synthetic strategy employed in the examples of the present invention are described in E. Atherton & R.C. Sheppard, "Solid Phase Peptide Synthesis: A Practical Approach", 1989, IRL Press, Oxford.

For example, use a resin with an acid-labile linker group in which the desired amino-protected C-terminal amino acid residue has been esterified. The amino protecting group is then removed and the second amino acid in the sequence is coupled using a suitable condensation reagent. Amino acids with semi-permanent amino protecting groups and functional side chains with permanent protecting groups are used. Amino-deprotection and coupling cycles are then repeated in alternating steps until assembly of the sequence of interest is complete.

Alternatively, said peptide V can be synthesized by solution-phase peptide synthesis methods known in the art, from the carboxy-terminus in a stepwise manner and/or by applying fragment condensation or ligation methods, using comprehensive or minimal protection strategies. A combined liquid-solid phase fragment condensation method can also be applied.

Typically, reactive side chain groups present (eg amino, hydroxyl, guanidino and carboxyl groups) will be protected throughout the synthesis as described above. A number of amino acid protecting groups are known from which to choose (see Grene, T.W. & Wuts, P.G.M (1991) Protecting Groups in Organic Synthesis, John Wiley & Sons, New York). Amino protecting groups that may be employed include 9-fluorenylmethoxycarbonyl (Fmoc) and tert-butoxycarbonyl (Boc). Possible side chain protecting groups include tert-butyl (tBu), trityl (Trt), Boc, and 2,2,5,7,8-pentamethylchroman-6-sulfo Acyl (Pmc). It should be appreciated that a wide variety of other such groups are known in the art.

Finally, the permanent side chain protecting groups are removed and the peptide is cleaved from the resin, usually simultaneously by treatment with a suitable acidic reagent, such as trifluoroacetic acid (TFA).

Conjugation of L and V:

L and V can be conjugated using all known methods of chemical synthesis. Particularly effective are nucleophilic substitution reactions in which a leaving group at the N-terminus of the peptide is replaced by a nucleophilic group on L. Such leaving group may be bromine attached alpha to the carbonyl, and the nucleophile may be nitrogen.

Conjugate Z to V or to L:

Z can be directly conjugated to V using the same method as L is conjugated to V. In the case where Z and V are linked through L, any chemical synthesis method can be used in the conjugation of Z and L. Particularly effective is the formation of amido bonds.

Example:

Example 1 Ang II analogs with optical imaging dyes

Fluorescein-NH-(CH ₂ ) ₂ -Gly-Arg-Val-Tyr-Ile-His-Pro-Ile-OH

Peptide analogs of Ang II were synthesized starting from 0.1 mmol Fmoc-Ile-Wang resin on an Applied Biosystems 433A peptide synthesizer. An excess of 1 mmol of preactivated amino acid (O-benzotriazol-1-yl-N,N,N',N'-tetramethyluronium hexafluorophosphate ( HBTU)). The N-terminus was bromoacetylated with 0.5 mmol bromoacetic anhydride in DMF for 30 minutes. The bromoacetylated resin was then treated with a solution of 0.5 mmol N-Boc-ethylenediamine dissolved in N-methylpyrrolidone (NMP) for 30 minutes. Peptides were cleaved from the resin with simultaneous removal of side chain protecting groups in 5 mL of TFA containing 2.5% triisopropylsilane (TIS) and 2.5% water for two hours. TFA was removed in vacuo, ether was added to the residue and the precipitated peptide was washed with ether and air dried.

23 mg of crude peptide, 16 mg of fluorescein NHS ester and 12 μL of N-methylmorpholine (NMM) were dissolved in dimethylformamide (DMF), and the reaction mixture was stirred overnight. The resulting reaction product was passed through preparative RP-HPLC (conditions: 5-50% B within 40 minutes, wherein A=H ₂ O/0.1% TFA and B=CH ₃ CN/0.1% TFA; flow rate, 10ml/min; column, Phenomenex Luna 5 μC18 (2) 250×21.20 mm) was purified to obtain 19 mg of pure fluorescein-peptide conjugate. The peptide was analyzed by analytical HPLC (conditions: gradient, 5-50% B in 10 minutes, where A=H ₂ O/0.1% TFA and B=CH ₃ CN/0.1% TFA; flow rate, 1 mL/min; column, Phenomenex Luna 3μC18 (2) 50×4.6mm; detection, UV214nm; product retention time 8.91min) for analysis. Further product identification was performed by electrospray mass spectrometry (MH ⁺ calculated, 1355.6; MH ⁺ found, 1355.2).

In a similar manner, Cy5.5 NHS ester (a dye supplied by Amersham Bioscience) can be used to synthesize the dye-peptide conjugate Cy5.5-NH-(CH ₂ ) ₂ -Gly-Arg-Val-Tyr- Ile-His-Pro-Ile-OH.

Example 2 Ang II analogs with ¹⁸ F for PET imaging

¹⁸ F-(CH ₂ ) ₃ -S-CH ₂ CO-NH-(CH ₂ ) ₂ -Gly-Arg-Val-Tyr-Ile-His-Pro-Ile-OH

ATII peptide analogs were synthesized starting from 0.1 mmol Fmoc-Ile-Wang resin on an Applied Biosystems 433A peptide synthesizer. An excess of 1 mmol of preactivated amino acid (using HBTU) was used in the coupling step ending at arginine. The N-terminus was bromoacetylated with 0.5 mmol bromoacetic anhydride in DMF for 30 minutes. The bromoacetylated resin was then treated with a solution of 0.5 mmol N-Boc-ethylenediamine dissolved in NMP for 30 minutes.

Peptides were cleaved from the resin with simultaneous removal of side chain protecting groups in 10 mL of TFA containing 2.5% TIS and 2.5% water for two hours. TFA was removed in vacuo, ether was added to the residue and the precipitated peptide was washed with ether and air dried.

The resulting crude peptide was treated with one equivalent of chloroacetic anhydride in DMF for 30 minutes. The resulting chloroacetylated peptides were purified using preparative RP-HPLC. Conjugation of F ¹⁸ -propylthiol to purified chloroacetylated peptides yields ¹⁸ F-labeled peptides for PET imaging.

Example 3 ^99m Tc-labeled Ang II analogs

cPn216-Gly-Arg-Val-Tyr-Ile-His-Pro-Ile-OH

Ang II peptide analogs were synthesized starting from 0.1 mmol Fmoc-Ile-Wang resin on an Applied Biosystems 433A peptide synthesizer. An excess of 1 mmol of preactivated amino acid (using HBTU) was used in the coupling step ending at arginine. The N-terminus was bromoacetylated with 0.5 mmol bromoacetic anhydride in DMF for 30 minutes. The resulting bromoacetyl resin was then treated with a solution of 0.2 mmol cPn216 and 0.4 NMM dissolved in DMF for 16 h.

Peptides were cleaved from the resin with simultaneous removal of side chain protecting groups in 5 mL of TFA containing 5% TIS, 5% water and 2.5% phenol for two hours. TFA was removed in vacuo, ether was added to the residue and the precipitated product was washed with ether and air dried.

Preparative RP-HPLC (0-30% B within 40 minutes, where A=H ₂ O/0.1% TFA and B=CH ₃ CN/0.1% TFA, on a Phenomenex Luna 5μC18(2) 250×21.20mm column The resulting product was purified to obtain 12 mg of pure chelated peptide conjugate. The resulting product was analyzed by analytical HPLC (conditions: gradient, 5-50% B in 10 minutes, where A = H ₂ O/0.1% TFA and B = CH ₃ CN/0.1% TFA; flow rate, 2 mL/min; column, Phenomenex Luna 3μC18 (2) 50×4.6mm; detection, UV214nm; product retention time 7.02min) for analysis. Further product identification was performed by using electrospray mass spectrometry (MH ⁺ calculated, 1280.8; MH ⁺ found, 1280.5)

Example 4 Ang II analogs for ^99m Tc labeling

cPn216-CO-(CH ₂ ) ₃ CO-NH-(CH ₂ ) ₂ -Gly-Arg-Val-Tyr-Ile-His-Pro-Ile-OH

Ang II peptide analogs were synthesized starting from 0.15 mmol Fmoc-Ile-Wang resin on an Applied Biosystems 433A peptide synthesizer. An excess of 1 mmol of preactivated amino acid (using HBTU) was used in the coupling step ending at arginine. The N-terminus was bromoacetylated with 0.75 mmol bromoacetic anhydride in DMF for 30 minutes. The resulting resin was then treated with a solution of 0.75 mmol of N-Boc-ethylenediamine dissolved in NMP for 60 minutes. Peptides were cleaved from the resin with simultaneous removal of side chain protecting groups in 10 mL of TFA containing 2.5% TIS and 2.5% water for 90 minutes. TFA was removed in vacuo, ether was added to the residue and the precipitated product was washed with ether and air dried.

5.5 mg of peptide, 23 mg of cPn216 tetrafluorothiophenylesters and 10 μL of NMM were dissolved in DMF and the resulting reaction mixture was stirred for 3 hours. The resulting product was analyzed by preparative RP-HPLC (conditions: 0-30% B within 40 minutes, where A=H ₂ O/0.1% TFA and B=CH ₃ CN/0.1% TFA; flow rate, 10 mL/min; column, Phenomenex Luna 5 μC18 (2) 250 x 21.20 mm) was purified to yield 4.3 mg of pure chelate-linker-peptide conjugate. The product was analyzed by analytical HPLC (conditions: gradient, 5-50% B over 10 minutes, where A=H ₂ O/0.1% TFA and B=CH ₃ CN/0.1% TFA; flow rate, 0.3 mL/min; column, Phenomenex Luna 3μC18 (2) 50×2mm; detection, UV214nm; product retention time 6.51min) for analysis. Further product identification was performed by using electrospray mass spectrometry (MH ⁺ calculated, 1436.9; MH ⁺ found, 1436.7)

Example 5 Antiarrhythmic Effects in a Pig Model After Myocardial Infarction (MI)

The drug cPn216-Gly-Arg-Val-Tyr-Ile-His-Pro-Ile-OH (X) was tested by the following procedure:

One pig was anesthetized and placed in artificial overpressure ventilation. The heart was approached and the anterior pericardium removed through a midline sternotomy. The LAD artery was closed by ligation on the second branch and one of the side branches was also closed as such. This results in frequent ventricular extrasystoles that progress to ventricular tachycardia and fibrillation within minutes from previous experiments with the same type of specimen. Within minutes after intravenous injection of 0.5 mg of the compound of formula (X), the symptoms were completely stabilized and sinus rhythm was restored. At this point notice that the average blood pressure starts to rise. Ventricular extrasystoles gradually reappeared after about 15 minutes and another injection of 0.5 mg of compound of formula (X) was given. The symptoms were again stable with a regular sinus rhythm. Over the next hour, a stable sinus rhythm and slightly decreased but essentially constant blood pressure were recorded. The pig was then given a lethal dose of potassium chloride to terminate the experiment. Analysis of ECG recordings before and after the first injection of compound (X) revealed a shortening of the QRS complex duration, suggesting that the antiarrhythmic effect may be related to an increase in myocardial conduction rate.

A similar set of observations was made in a second pig experiment.

Thus, the compound of formula (X) was shown to be useful as an antiarrhythmic drug in this specimen.

Example 6 Synthesis of N-((CH ₂ ) ₆ -tetraamine)-Gly-Arg-Val-Tyr-Ile-His-Pro-Ile-OH

Starting from 0.25 mmol Fmoc-Ile-Wang resin, the Arg-Val-Tyr-Ile-His-Pro-Ile peptide sequence was synthesized on an Applied Biosystems 433A peptide synthesizer using the Fmoc/tert-butyl strategy. An excess of 1 mmol of amino acid preactivated with O-benzotriazol-1-yl-N,N,N',N'-tetramethyluronium hexafluorophosphate (HBTU) was used in the coupling step. The free N-terminal was bromoacetylated with 1 mmol bromoacetic anhydride in dimethylformamide for 30 minutes. The resulting bromoacetylated resin (0.05 mmol) was subsequently treated with a solution of 0.15 mmol of the tetra-Boc-tetramine chelate dissolved in dimethylformamide for 60 minutes.

The peptide was cleaved from the resin (0.025 mmol) with simultaneous removal of side chain protecting groups by treatment with 5 mL of trifluoroacetic acid containing 2.5% triisopropylsilane and 2.5% water for 90 minutes. The trifluoroacetic acid was removed in vacuo, diethyl ether was added to the residue and the precipitate was washed with diethyl ether and air-dried to give 25 mg of crude product.

The 25 mg crude product was analyzed by preparative RP-HPLC (0-30% B within 40 minutes, where A=H ₂ O/0.1% TFA and B=CH ₃ CN/0.1% TFA, in a Phenomenex Luna 5μC18(2) 250× 21.20mm column flow rate is 10mL/min) for purification to obtain 6.5mg semi-purified product. The second purification step of the semi-purified product (A=H ₂ O/0.1% HCOOH and B=CH ₃ CN/0.1% HCOOH, and the rest were the same conditions as above) yielded 3 mg of pure product. The product was analyzed by HPLC (conditions: gradient, 0-30% B within 10 minutes, where A=H ₂ O/0.1% HCOOH and B=CH ₃ CN/0.1% HCOOH; flow rate, 0.3 mL/min; column, Phenomenex Luna 3μC18 (2) 50×2mm; detection, UV214nm; product retention time, 5.34min) for analysis. Further product identification was performed by electrospray mass spectrometry (MH ⁺ calculated, 1196.8; MH ⁺ found, 1196.7)

Example 7 Hcy3-5; Synthesis of N-cPn216-Gly-Arg-Hcy-Tyr-Hcy-His-Pro-Ile-OH

Starting with 0.25 mmol Fmoc-Ile-Wang resin, the Arg-Hcy-Tyr-Hcy-His-Pro-Ile peptide sequence was synthesized on an Applied Biosystems 433A peptide synthesizer using the Fmoc/tert-butyl strategy. An excess of 1 mmol of amino acid preactivated with O-benzotriazol-1-yl-N,N,N',N'-tetramethyluronium hexafluorophosphate (HBTU) was used in the coupling step. The free N-terminal was bromoacetylated with 1 mmol bromoacetic anhydride in dimethylformamide for 30 minutes. The resulting bromoacetylated ^resin (0.05 mmol) was subsequently treated with a solution of 0.25 mmol of cPn216 ( ^99m Tc-chelating agent ¹ ) (see patent WO20030049 for details of the synthesis) dissolved in dimethylformamide for 60 minutes.

The peptide was cleaved from the resin and the side chain protecting groups were simultaneously removed by treatment in 10 mL of trifluoroacetic acid containing 2.5% triisopropylsilane and 2.5% water for 2 hours and 20 minutes. Trifluoroacetic acid was removed in vacuo, diethyl ether was added to the residue and the precipitate was washed with diethyl ether and air-dried to give 70 mg of crude linear product.

The crude linear product was analyzed by preparative RP-HPLC (5-50% B within 40 minutes, where A=H ₂ O/0.1% TFA and B=CH ₃ CN/0.1% TFA, on a Phenomenex Luna 5μC18(2) 250× 21.20mm column flow rate is 10mL/min) for purification to obtain 20mg of pure linear product. The linear product was dissolved in 2 mL DMSO and 200 mL water, and the resulting solution was adjusted to pH 8 with aqueous ammonia. The resulting solution was stirred for 26 hours and then adjusted to pH 2 with trifluoroacetic acid.

The resulting cyclic product was purified by preparative RP-HPLC (0-30% B, the rest were the same conditions as above) to obtain 12 mg of pure cyclic product. The resulting product was analyzed by HPLC (conditions: gradient, 5-50% B in 10 minutes, where A = H ₂ O/0.1% TFA and B = CH ₃ CN/0.1% TFA; flow rate, 0.3 mL/min; column, Phenomenex Luna 3μC18 (2) 50×2mm column; detection, UV214nm; product retention time, 6.01min) for analysis. Further product identification was performed by electrospray mass spectrometry (MH ⁺ calculated, 1300.7; MH ⁺ found, 1300.6).

Sequence Listing

<110>Amersham Health AS

<120>Angiotensin II analogues

<130>PN0380

<160>5

<170>PatentIn version 3.1

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<213> Artificial sequence

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<223> Arg, N-alkylated Arg, or Arg mimics

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<223> Amino Acids Containing Hydrophobic Side Chains

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Xaa Xaa Val Tyr Ile His Pro Xaa

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<223>Bip-2-amino-3-biphenylpropionic acid

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Claims (11)

1. A medicine characterized by formula (I) Z-(L) _n -V (I) in, V represents a peptide with the binding sequence -X ¹ -X ² -Val-Tyr-Ile-His-Pro-X ³ , L indicates an optional linker, Z represents a group which may optionally carry an imaging moiety M, n is 0 or 1, X ¹ represents an amino acid, ^X2 represents Arg or N-alkylated Arg or Arg mimics, X ³ represents an amino acid comprising a hydrophobic side chain, wherein the Val and Ile residues at positions 3 and 5 are each optionally replaced by an amino acid capable of forming a bridge, Z is optionally bonded to amino acid ^X through a linker L, and M, when present, represents an imageable moiety, which can be detected directly or indirectly during a diagnostic imaging procedure. 2. The medicament according to claim 1, which is effective in the treatment of heart failure, arrhythmia and other fibrotic prominent diseases and in the treatment of COPD, liver fibrosis and arteriosclerosis. 3. The medicament according to claim 1, for use in diagnosis, wherein M is an in vivo imageable moiety. 4. The medicine of any one of claims 1-3, wherein Z represents a chelating agent of formula (VII) in: Each of R ¹ , R ² , R ³ and R ⁴ is independently H or C _1-10 alkyl, C _3-10 alkylaryl, C _2-10 alkoxyalkyl, C _1-10 hydroxyalkyl , C _1-10 alkylamine, C _1-10 fluoroalkyl, or 2 or more R groups together with the atoms connecting them form a saturated or unsaturated carbocyclic or heterocyclic ring. 5. The medicine of any one of claims 1-4, wherein Z represents a chelating agent of formula (XI)

Wherein Q ₁ -Q ₆ are independently Q groups, wherein Q is H, alkyl, aryl or amine protecting group, W ₁ is -NR-, -CO ₂ -, -CO-, -NR(C=S)-, -NR(C=O)-, -CONR- or Q group; each Y is independently a D- or L-amino acid, -CH ₂ -, -CH ₂ OCH ₂ - or -OCH ₂ CH ₂ O-, or an X group; p is an integer whose value is 1-8; q is an integer whose value is 0-30; R is H, C _1-4 alkyl, C _2-4 alkoxyalkyl, C _1-4 hydroxyalkyl or C _1-4 fluoroalkyl; Q is A is a counter ion. 6. The medicine according to any one of claims 1 and 3-5, wherein M represents a gamma radiation moiety for Radio or SPECT imaging, comprising ⁶⁷ Ga, ¹¹¹ In, ¹²³ I, ¹²⁵ I, ¹³¹ I, ^81m Kr, ⁹⁹ Mo, ^99m Tc, ²⁰¹ Tl and ¹³³ Xe. 7. The medicament according to the preceding claims, for use in therapy, having formula (X) or formula (XII)

Formula (X) Formula (XII) Or used as a diagnostic agent, having formula (Xa) or (XIIa)

8. A pharmaceutical formulation comprising a compound of formula (I) as claimed in claim 1 and one or more pharmaceutically acceptable additives and/or excipients. 9. The use of the medicament according to claim 1, said use is for treating and/or diagnosing heart failure, arrhythmia and other diseases with prominent fibrosis, especially diseases such as COPD, liver fibrosis and arteriosclerosis. 10. An in vivo diagnostic method for heart failure and other fibrosis-prominent diseases of a subject, especially COPD, liver fibrosis and arteriosclerosis, said method comprising administering the formula (I) described in claim 1 Drugs and subsequent partial or whole-body imaging of the subject. 11. A kit for the preparation of a radiopharmaceutical composition of formula (I), said kit comprising a peptide-chelate conjugate and a reducing agent.

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