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 an imaging moiety

Technical Field

The present invention provides novel drugs effective in the treatment of heart failure, arrhythmia and other diseases in which fibrosis is prominent and in the diagnosis of diseases and conditions in which the fibrotic process is prominent. The invention also provides novel pharmaceutical compositions and precursors for the preparation of diagnostic agents. In addition, the present invention provides novel drugs that are effective in monitoring treatment and methods of monitoring treatment. Still further objects of the invention are apparent in the claims.

The novel drugs comprise a targeting moiety that binds to a receptor that may or may not be up-regulated and/or over-expressed in the disease region. The targeting moiety comprises an imaging agent group with a diagnostically imageable moiety, an optional linker group, and a peptide moiety.

The novel pharmaceutical compounds are useful for angiotensin receptors (angiotensin receptors), especially for angiotensin II (Ang II) type 1 (AT)₁) The receptors have high affinity.

Background

Angiotensin ii (ang ii) -octapeptide (Asp-Arg-Val-Tyr-Ile-His-Pro-Phe) -is a pleiotropic vasoactive peptide that binds to two different receptors: ang II1 type (AT)₁) And type 2 (AT)₂) The receptor binds. Activation of the renin-angiotensin-aldosterone system (RAAS) can lead to vascular hypertrophy, vasoconstriction, salt and water retention, and hypertension. These effects are mainly attributed to AT₁Receptor mediation. Paradoxically, other Ang II-mediated effects, including cell death, vasodilation, and natriuresis, are mediated by AT₂Receptor activation mediated. The mechanism of Ang II signaling is not fully understood. AT₁Receptor activation triggers a variety of intracellular systems including tyrosine kinase-induced protein phosphorylation, production of arachidonic acid metabolites, activity modification of reactive oxidant species, and intracellular Ca⁺The concentration fluctuates. AT₂Receptor activation leads to bradykinin stimulation, nitric oxide production, and prostaglandin metabolism, which are in large part related to AT₁The receptor action is reversed. (see: BerryC. Touyz R, dominicak AF, Webb RC, Johns DG.: Am J Heart Circuit Physio.2001Dec; 281 (6): H2337-65. angiotensin receptors: signaling, vascular pathophysiology and interaction with ceramides).

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

The RAAS system is illustrated herein in FIG. 1, which is Foote et al Ann.27: FIG. 1 of the literature is shown in 1495-1503 (1993).

In addition to its important role in normal cardiovascular homeostasis, the overactivity of RAAS is also associated with the development of a number of cardiovascular diseases such as hypertension, congestive heart failure, coronary ischemia and renal insufficiency. Following Myocardial Infarction (MI), RAAS is activated. Especially AT₁The receptor appears to play a major role in post-MI remodeling, as AT is post-MI and left ventricular dysfunction₁Expression of the receptor is increased. Thus, those agents that interfere with RAAS, such as ACE inhibitors and AT₁Receptor antagonists have been shown to have a powerful therapeutic effect in such cardiovascular diseases.

For the heart, kidneys, lungs, liver, etc., fibrosis is a common factor in the failure of these organs. There is therefore considerable interest in understanding the pathophysiological mechanisms involved in organ fibrosis, especially in making possible protective pharmacological strategies. Tissue repair involves inflammatory cells, including members of the monocyte/macrophage lineage, which collectively initiate the repair process; and myofibroblasts (myofibrasts), phenotypically transformed mesenchymal fibroblasts, responsible for collagen turnover and fibrous tissue formation. Each of these cells present in the repair microenvironment is associated with a regeneration leading to angiotensinogen II (Ang II)Is related to the molecular event(s). In the autocrine/paracrine mode, these peptides pass through Angiotensinogen (AT)₁) Receptor ligand binding regulates expression of TGF- β 1. It is this cytokine that changes the fibroblast phenotype to myofibroblasts (myoFb) and regulates collagen turnover by myofibroblasts. Angiotensin-converting enzyme (ACE) inhibition or AT₁Receptor antagonism has been considered a protective intervention since it individually blocks many of these molecular and cellular responses leading to fibrosis. (see: Weber KT. Fibrosis, a common to organic failure: angiotensin II and dtissure repair. Semin Nephrol.1997 Sep; 17 (5): 467-91and preferences thein).

Ang II can regulate tissue fibrosis by activating stromal cells. For example, Ang II is activated in vitro by AT₁Stimulating the proliferation of cardiac fibroblasts. AT has been shown in vitro on cardiac fibroblasts₁The presence of a receptor. Most of the pro-fiber (profibrotic) actions of Ang II appear to be mediated through this receptor; however, AT has been detected on a hypertrophied human heart₂Expression is increased on cardiac fibroblasts, and the balance between expression of these two subtypes may be critical for determining Ang II responses. (see: am. J. Respir. crit. Care Med., Volume June 2000, 1999 2004Angiotensin II is Mitogenic for human Lung fibers Activation of the Type 1Receptor RICHARD P. MARSHALL, ROBIN J. MCANNULTY, and GEOFFREY J. LAURENTAND refer heat).

The Ang II receptor can be distinguished by the inhibitory effect of a particular antagonist. AT₁Receptors are selectively antagonized by biphenylimidazoles, such as Losartan (Losartan), while tetrahydroimidazopyridines (tetrahydroimidazopyridines) specifically inhibit AT₂A receptor. The AT₂The receptor may also be encoded by CGP-42112A (which is a hexapeptide analogue of AngII which also inhibits AT₂Receptor, depending on concentration) is selectively activated. Two other angiotensin receptors have been described: AT₃And AT₄The subtype is.

In rodents, AT₁The receptors have two functionally distinct subtypes, AT_1AAnd AT_1BAnd has amino acid sequence homology of more than 95%.

The second major angiotensin isoform (isoform) is AT₂A receptor. It and AT_1AOr AT_1BThe receptor has low amino acid sequence homology (-34%). Although AT₂The precise signaling pathways and functional roles of receptors remain unclear, but these receptors may antagonize AT under physiological conditions by inhibiting cell growth and by reducing apoptosis and vasodilation₁-mediated effects. AT₂The precise role of the receptor in cardiovascular disease remains to be defined.

Except for AT₁And AT₂In addition, other Ang II receptors are known and are commonly referred to as ATs_Atypical(see Kang et al, am. Heart J.127：1388-1401(1994))。

Inhibition of the Ang II effect has been used in medicine, for example in the control of hypertension and heart failure. This has been achieved in a number of ways: by using renin inhibitors which block the conversion of angiotensinogen to angiotensin I (the precursor of Ang II); by using angiotensin enzyme (ACE) inhibitors that block the conversion of angiotensin I to Ang II (and 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 anti-arrhythmic therapy. Antiarrhythmic drugs have limited overall success and calcium channel blockers can sometimes cause arrhythmias. No single drug showed superiority except for the possibility of amiodarone. Short-term antiarrhythmic benefits, depending on the specific drug, have been found to be offset by their neutral or negative effects on mortality. (Sanguinetti MC and Bennett, PB: antiarrhythmic drug target selection and screening. Circulation 2003, 93 (6): 491-. There is a clear need for better antiarrhythmic drugs.

The published article by Lancet (Llindholm, LH et al; sudden cardiac death effect of Losartan (Losartan) in diabetic patients: data from LIFE studies. The Lancet, 2003, 362: 619-₁In addition to being generally beneficial to patients with CHF, receptor antagonists also reduce the incidence of sudden cardiac death. There are several studies that suggest AT₁Receptor antagonists have antiarrhythmic effects on arrhythmias caused by myocardial infarction or reperfusion after LAD ligation (Harada K et al. angiotensin II1a type receptor is associated with reperfusion arrhythmia. circulation.1998, 97: 315-317.Ozer MK et al, Captopril and losartan effects in mouse myocardial ischemia-reperfusion induced arrhythmias and gangrene. Pharmacological research, 2002, 45(4), 257-263Lynch JJ et al, EXP3174, losartan antagonizes human metabolites in total but neither Canlosartan nor the angiotensin converting enzyme inhibitor captopril, avoiding lethal ischemic arrhythmias in recent myocardial infarction canine models JACC, 1999, 34876-seco 884).

Description of the Related Art

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

U.S. Pat. No. 4411881(New England Nuclear Corporation) describes the stabilization of radio-labeled compounds. Examples of radio-labeled compounds include, for example, angiotensin II (5-L-isoleucine) [ tyrosyl-¹²⁵I]- (monoiodination).

Ang II can be converted to potent antagonists or partial antagonists by making changes in their amino acid composition. For example, replacement of phenylamino propionic acid with isoleucine at position 8 and aspartic acid with sarcosine at position 1 would render the peptides effective antagonists.

And AT₁The affinity specificity of the receptor can be determined by substituting the 4-position of the amino acidAnd 6 position by cyclization or bridging. Similarly, the introduction of sarcosine AT position 1and glycine AT position 8 converted the peptide to AT₁Selective antagonists (see RC Speth. Sarcosinell, Glycine 8 angiotensin II is AT₁Angiotensin II receptor subtype selective antagonists Regulatory peptides 115(2003) 203-209).

As mentioned above, AT₁The natural ligand of the receptor is the octapeptide Ang II, Asp-Arg-Val-Tyr-Ile-His-Pro-Phe, which is in the nanomolar range with AT₁Receptor binding.

When a natural binding ligand is modified by combining a moiety with other moieties, particularly with relatively large and bulky moieties, the affinity of the peptide carrier is often compromised.

We have surprisingly found that the octapeptide AngII, Asp-Arg-Val-Tyr-Ile-His-Pro-Phe and derivatives thereof, when substituted AT specific positions on the peptide, not only retains its binding capacity but also surprisingly increases its ability to bind angiotensin II receptors, particularly for AT₁The affinity of the receptor.

Summary of The Invention

It is a first object of the present invention to provide a medicament useful for the treatment of heart failure, cardiac arrhythmias and other diseases where fibrosis is prominent, such as COPD, liver fibrosis and arteriosclerosis, comprising a targeting moiety shown for AT₁The receptor has a stronger affinity than the native octapeptide Ang II. The drug should exhibit antagonistic activity.

It is a second object of the invention to provide a medicament comprising a targeting moiety in combination with an imageable moiety which is effective in diagnosing heart failure and other diseases where fibrosis is prominent, such as COPD, liver fibrosis and arteriosclerosis. The imageable portion can be any imageable portion that, when administered to a subject, produces a copy of the subject at least where a portion of the contrast agent is distributedImaging, such as by radiographic, SPECT, PET, MRI, X-ray, Optical Imaging (OI), Ultrasound (US), electronic impedance, or magnetic imaging modalities. Targeting moiety-to-AT in combination with imageable forms₁The receptor should show a higher affinity than Ang II and should preferably be used as an antagonist, although weak agonist activity may still be acceptable.

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

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

The medicaments of the invention comprise a peptide V, optionally linker L and Z moieties, as shown in formula (I)

Z-(L)_n-V (I)

Wherein,

v represents a peptide having the binding sequence-X¹-X²-Val-Tyr-Ile-His-Pro-X³The peptide of (a) above (b),

l represents an optional linker, and L represents an optional linker,

z represents a group optionally carrying an imaging moiety M,

n is a number of 0 or 1,

X¹represents an amino acid, and represents a cyclic amino acid,

X²represents Arg or N-alkylated Arg, or a mimic of Arg,

X³refers to an amino acid comprising a hydrophobic side chain,

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

z is optionally joined to amino acid X via a linker L¹Bonded, and the presence of M represents an imageable moiety that can be detected directly or indirectly in a diagnostic imaging procedure.

Detailed Description

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

In the targeting moiety of formula (I), V as defined above represents the peptide sequence-X¹-X²-Val-Tyr-Ile-His-Pro-X³。

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

The three letter abbreviations 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-pentanoic acid

Nle-2-amino-hexanoic 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 said V are preferably independently selected

X¹represents-NY₁-(CH₂)_m-CO-wherein m is an integer from 1 to 10 and Y₁Is H or alkyl or aryl containing substituents, most preferably Gly

X²Represents Arg or N-methyl-Arg or Arg-mimetic Phe [ 4-guanidino ]]And Gly-4-piperidyl [ N-amidino group]，

X³Represents Phe, D-Phe, Ile, Abu, Nva, Nle, Phg; hph, Bip, Nal or Cha, most preferably D-Phe, Bip, Ile or Hph.

Preferably wherein X¹Denotes Gly, X²Represents Arg or N-methyl-Arg and X³Represents a drug of D-Phe, Bip, Ile or Hph.

Still further preferred is where X¹Denotes Gly, X²Represents Arg and X³A drug representing Ile.

If the amino acids in positions 3 and 5 are selected to form a bridge unit, the bridge preferably contains a-CH₂-CH₂-、-S-CH₂-、-S-CH₂-S-, lactam or-S-S-units. More preferably the covalent bond is a disulfide bond formed by the oxidation of 2 cysteine or homocysteine pairings.

Examples of suitable linkers L are described in patents WO 98/18496 and WO 01/77145 (pages 23-27), the contents of which are hereby incorporated by reference. L may preferably represent polyalkylene glycol units, such as polyethylene glycol (PEG) and polypropylene glycol (PPG), saccharides, dextran or 1-10 amino acids. The linker may also act as a biological modifier, as described in WO 03/006491.

The linker L may also be derived from an alkylamine or arylamine, preferably of the formula NH- (CH)₂)_mA compound of (a) optionally with-CO- (CH)₂)_m-CO-binding, wherein m represents a positive integer from 1 to 10.

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

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

formula (V)

Formula (VI)

The Z moiety comprises a non-peptidic moiety having a molecular weight of more than 50D (Dalton), more preferably between 100-1000D and even more preferably between 300-700D. Provided that the final targeting moiety of formula (I) is shown to be directed to AT₁The receptor has a higher affinity than the natural ligand Ang II, and the Z moiety can be any pharmaceutically acceptable chemical entity. More specifically, Z represents an organic group having 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 linear or branched hydrocarbon group optionally containing one or more double or triple bonds and optionally substituted with halogen, oxygen, sulphur or phosphorus atoms or optionally comprising heteroatoms such as oxygen, nitrogen or sulphur. More specifically, the hydrocarbyl group may represent a substituted or unsubstituted alkyl, alkenyl, alkynyl group having a molecular weight of at least 50D.

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

Z may further represent one or more attached heterocyclic compounds, such as a5, 6, 7, 8, 9 or 10-membered ring system, which may be monocyclic, bicyclic or tricyclic and may comprise one or more of N, O, S and P as heteroatoms. Such ring systems may also be linked to hydrocarbon and carbocyclic groups and are as defined above or fused to carbocyclic groups. Examples of such groups are acridinyl, benzofuranyl, indolyl, pyridyl, piperidyl, morpholinol and thienyl.

Z may also represent polyalkylene glycols, such as polyethylene glycol (PEG) and polypropylene glycol (PPG), all saccharides with a molecular weight of more than 50D, such as mono-or polysaccharides. Polyalkylene glycols can additionally be used as biological modifiers.

Specific Z denotes a chelating agent such as the acyclic or cyclic polyaminocarboxylates (e.g., DTPA-BMA, DOTA and DO3A) described in U.S. Pat. No. 4,647,447(Schering AG) and WO86/02841 (Nycomed Saluta, Inc.), which are incorporated herein by reference. Chelating agents also include amine thiols, such as diamine dithiols, aminated oximes (aminoximes), and hydrazines, and related reagents as described in patent WO 01/77145 (see table 1 therein), incorporated herein by reference. The chelating agent cPN216 of formula (VIII) is particularly preferred.

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

For drugs to be effective in diagnosis, especially in vivo diagnosis, the Z moiety must be capable of carrying an imageable moiety or moiety designated by M. With is meant any form of linkage between the Z and M moieties, such as a chemical bond, such as a covalent or electro-valent bond or an ionic bond, or by adsorption or any other type of linkage.

Z may be any imageable moiety. Wherein M denotes a metal entity, Y₁Represents a chelating agent. Z and/or Y₁The nature of M will depend on the imaging modality used in the diagnosis. Z and/or Y₁M must be capable of being detected directly or indirectly during in vivo diagnostic imaging procedures, such as moieties that emit or can cause emission of detectable radiation (e.g., by radioactive decay, fluorescence excitation, spin resonance excitation, etc.), moieties that affect local electromagnetic fields (e.g., paramagnetic, superparamagnetic, ferrimagnetic, or ferromagnetic species), moieties that absorb or scatter radiation energy (chromophores, particles (including vesicles containing gases or liquids), heavy elements and compounds thereof, etc.), and moieties that produce detectable species (e.g., gas microbubble generators).

In a preferred embodiment, one Z moiety is reacted with X¹Directly covalently bonded to form an N-alkylglycine unit.

The chelating agents of the formulae (VII) and (VIII) below 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 moieties Z and M are described in more detail below:

in a first embodiment, the compounds of formula (I) comprise Y with one or more imageable moieties M effective in both Radio and SPECT imaging modalities₁And (4) partial. 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 radionuclides⁶⁷Ga、¹¹¹In、¹²³I、¹²⁵I、¹³¹I、^81mKr、⁹⁹Mo、^99mTc、²⁰¹Tl and¹³³xe. Most preferably^99mTc。

M may be further represented by the following isotopes or isotope pairs for imaging and therapy without the need to alter the radiolabeling methodology or chelating agents:⁴⁷Sc₂₁；¹⁴¹Ce₅₈；¹⁸⁸Re₇₅，¹⁷⁷Lu₇₁；¹⁹⁹Au₇₉；⁴⁷Sc₂₁；¹³¹I₅₃；⁶⁷Cu₂₉；¹³¹I₅₃and¹²³I₅₃；¹⁸⁸Re₇₅and^99mTc₄₃；⁹⁰Y₃₉and⁸⁷Y₃₉；⁴⁷Sc₂₁and⁴⁴Sc₂₁；⁹⁰Y₃₉and¹²³I₅₃；¹⁴⁶Sm₆₂and¹⁵³Sm₆₂(ii) a And⁹⁰Y₃₉and¹¹¹In₄₉。

when M represents a metal radionuclide, then Y₁Denotes 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 the formula (VII) are particularly preferred:

wherein:

each R¹、R²、R³And R⁴Independently is H or C_1-10Alkyl radical, C_3-10Alkylaryl group, C_2-10Alkoxyalkyl group, C_1-10Hydroxyalkyl radical, C_1-10Alkylamine, C_1-10Fluoroalkyl, or 2 or more R groups, together with the atoms linking them, form a saturated or unsaturated carbocyclic or heterocyclic ring.

More particularly preferred chelating agents are of formula (VII), wherein R¹、R²And R³Is hydrogen or methylAnd R is⁴Is an alkylamine group, most particularly a compound of formula (VIII), herein denoted as cPN 216.

Formula (VIII)

For Y₁Most preferably, when the chelator is cPN216, the imaging factor M is^99mTc。

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

Other preferred chelating agents are compounds of formula (XI)

Formula (XI)

Wherein Q₁-Q₆Is an independent Q group, wherein Q is H, alkyl, aryl, or an amine protecting group.

W₁is-NR-, -CO₂a-CO-, -NR (C ═ S) -, -NR (C ═ O) -, -CONR-or Q group;

each Y is independently a D-or L-amino acid, -CH₂-、-CH₂OCH₂-or-OCH₂CH₂An O-or X group;

p is an integer having a value of 1 to 8;

q is an integer having a value of 0 to 30;

r is H, C_1-4Alkyl radical, C_2-4Alkoxyalkyl group, C_1-4Hydroxyalkyl or C_1-4A fluoroalkyl group;

q is

A is a counter ion;

the synthesis of tetraamine chelators of formula (XI) can be found in british patent application No. GB 0416062.8.

Non-metallic radionuclides, e.g.¹²³I、¹²⁵I and¹³¹i may be either with the L moiety (when L is present) or with X₁Covalently linked 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 PET imaging modalities. Z represents a radiation emitter with positron emission properties. Preferred Z groups are radionuclides¹¹C、¹⁸F、⁶⁸Ga、¹³N、¹⁵O and⁸²Rb。¹⁸f is particularly preferred. Metal radiation emitter⁸²Rb and⁶⁸ga and chelating agent Y₁Chelation is also preferred.

Thiol coupling chemistry,¹⁸The F-synthons and labeled peptides were prepared using thiol coupling chemistry as described in WO03/080544, the contents of which are incorporated herein by reference.

A description of labelling peptides by using thiol coupling chemistry can be found in uk patent application No. 0317815.9, the contents of which are incorporated herein by reference.

When M represents a metal radionucleus, then Y₁Denotes a chelating agent suitable for forming a stable chelate with M. Such chelators are well known in the art at this stage, typical examples of which are described in table 1 of WO 01/77145 and previously in the Radio and SPECT imaging sections.

In another preferred embodiment, Y₁Is a DOPT chelator and M is⁶⁸Ga, which can be easily introduced into the chelate using microwave chemistry.

Non-metallic radionuclides such as¹⁸F, when present, may be covalently linked to the L moiety, or may be reacted with X by substitution or addition reactions known in the art₁Ligation, these reactions are also described in WO03/080544, which is hereby incorporated by reference.

In a third embodiment, the compounds of formula (I) comprise Y with one or more M imageable moieties that are effective in MR imaging modalities₁And (4) partial. M here denotes a paramagnetic metal, Gd, as described in U.S. Pat. No. 4647447³⁺、Dy³⁺、Fe³⁺And Mn²⁺Is particularly preferred and Y₁Denotes chelating agents, in particular acyclic or cyclic polyaminocarboxylates (e.g. DTPA, DTPA-BMA, DOTA and DO3A) as described in U.S. Pat. No. 4647447 and WO 86/02841. M may also represent a metal oxide absorbed by Z, such as a superparamagnetic, ferrimagnetic or ferromagnetic species, so that Z acts as a coating for the metal oxide. Metal oxides for use as MR contrast agents are described in us patent 6223777, which is hereby incorporated by reference.

In a fourth embodiment, the compound of formula (I) comprises Y with one or more M moieties that are imageable in X-ray imaging modalities₁And (4) partial. M here denotes a heavy metal, such as W, Au and Bi, preferably in the form of an oxide that can be absorbed by Z. Z may also be represented by iodinated aryl derivatives, in particular those known as X-ray contrast agents, such as Iopamiron^TMAnd Omnipaque^TM. These agents can be linked to the peptide V of formula (I) via their amide or amine functions.

In another embodiment, the compound of formula (I) comprises Z in the form of gas-filled vesicles. Such ultrasound imaging agents may be used in receptor imaging, such as when they are used in conjunction with peptides, as described in the art, such as in WO 98/18500.

In a sixth embodiment of the invention, the moiety Z of formula (I) may be any moiety that can be detected directly or indirectly in an optical imaging procedure. The detectable moiety may be a light scatterer (e.g., 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 interacts with light in the electromagnetic spectrum with wavelengths from ultraviolet to near infrared. In a preferred version, Z has fluorescent properties.

Preferred organic dye moieties include groups having a broad delocalized electron system, such as cyanines (cyanines), merocyanines (merocyanines), indocyanines (indocyanines), phthalocyanines (phthalocyanines), naphthalocyanines (naphthalocyanines), triphenylmethines (triphenylmethines), porphyrins (porphyrins), pyrylium (pyrilium) dyes, thiapyrylium (thiapyrilium) dyes, squarylium dyes, croconium dyes, azulenium dyes, indoanilines (indoanilides), benzophenoxazinium (benzoxazinium) dyes, benzothiophene (benzothiophenium) dyes, anthraquinones (anthraquinones), naphthoquinones (naphthoquinones), indanthrenes (chromenes), thiochromenones (dithiothrene complexes), thiochromenones (thiocyanines), thiochromenones (thiochromenones), thiochromenones (thiochromeno, Iodoaniline dyes, bis (S, O-dithiole) complexes. 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 (e.g., europium, samarium, terbium or dysprosium) are used in some cases, such as fluorescent nanocrystals (quantum dots).

Further description of suitable parts in optical imaging procedures can be found in norwegian patent application No. 200303115, the contents of which are incorporated herein by reference.

The drug of formula (I) may be further modified by the attachment of one or more groups of biological modifiers, such as polyalkylene glycol units, e.g. polyethylene glycol (PEG) and polypropylene glycol (PPG). Examples of biological modifier groups are described in WO03/006491, the contents of which are incorporated herein by reference. The biological modifier group may be attached at any position in the compound of formula (I) as long as it does not have a significant negative effect on the attachment of the compound to the targeted receptor.

The biological modifier is preferably based on a monodisperse PEG building block and comprises 1-10 building block units, and has the function of changing the pharmacokinetics and plasma clearance rate of the medicament. In a preferred embodiment of the invention, the compound of formula IV, represents a unit of a biological modifier consisting of a polymer of monodisperse PEG-like structure, 17-amino-5-oxo-6-aza-3, 9, 12, 15-tetraoxaheptadecanoic acid.

Wherein n is an integer from 1 to 10 and wherein the C-terminal unit forms an amide bond.

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

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

Z-CO-NH-(CH₂)_m-X¹-X²-Val-Tyr-Ile-His-Pro-X³(II)

Z-CO-(CH₂)_m-CO-NH-(CH₂)_m-X₁-X₂-Val-Tyr-Ile-His-Pro-X₃(III)

wherein m is an integer between 1and 5, and Z, X¹、X²And X³As defined hereinbefore.

A compound defined by formulas Va and VIa:

wherein Z and V are as defined above.

A compound of formula (IX):

wherein Y is²Is alkyl, aryl or a moiety comprising a short PEG and Y²Is preferably-CH₂-CH₂-CH₂And X³As defined above.

Preferred examples of the drug of formula (I) are represented by the following:

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-((CH₂)₆-tetramine) -Gly-Arg-Val-Tyr-Ile-His-Pro-Ile-OH

As noted above, for purposes of use in vivo diagnostics, the band^99mTc or¹⁸Chelates of compounds of F are particularly preferred.

Particularly preferred is the compound cPN216-Gly-Arg-Val-Tyr-Ile-His-Pro-Ile-OH, the chemical structure of which is represented by formula (X):

formula (X)

And it and^99mtc chelate compounds.

Particular preference is given to the compound N- ((CH)₂)₆-tetramine) -Gly-Arg-Val-Tyr-Ile-His-Pro-Ile-OH, the structural formula of which is represented by formula (XII):

formula (XII)

And it and^99mtc chelate compounds.

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 by means of a formulation suitable for injection or infusion, such as an aqueous solution. The formulation may contain one or more pharmaceutically acceptable additives and/or excipients, such as buffers; solubilizers such as cyclodextrins; or a surfactant such as Pluronic (Pluronic), Tween (Tween) or a phospholipid. Additionally, a stabilizer or antioxidant such as ascorbic acid, gentisic acid or p-aminobenzoic acid and a bulking agent such as sodium chloride or mannitol for lyophilization can be added.

In one aspect of the invention, the agents of formula (I) are useful in therapy as well as in therapy monitoring. In the monitoring of the progression of treatment of heart failure and other diseases in which fibrosis is prominent, in particular COPD, liver fibrosis and arteriosclerosis, one of the methods is to administer to the subject to be treated a drug of formula (I) and subsequently generate an image of the whole body of the subject or of a part of the subject.

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

General procedure for the preparation of drugs and their precursors

The abbreviations used have the following meanings:

fmoc: 9-Fluorenylmethoxycarbonyl

Boc: tert-butoxycarbonyl group

tBu: tert-butyl radical

Trt: trityl radical

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

TFA: trifluoroacetic acid

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

DMF: dimethyl formamide

NMP: n-methyl pyrrolidone

And (3) TIS: tri-isopropyl silyl

NHS: n-hydroxysuccinimide radical

NMM: n-methylmorpholine

RP-HPLC: reversed phase high performance liquid chromatography

Wang resin: P-Phenylmethoxybenzyl alcohol resin

Synthesis of V:

the peptide V described in the present invention can be synthesized by employing all known chemical synthesis methods but is particularly effective by the Merrifield solid phase method using an automated peptide synthesizer (J.Am.chem.Soc., 85: 2149 (1964)). Typically, the desired sequence is assembled by solid phase peptide synthesis. Standard procedures for synthetic strategies employed in the examples of the invention are e.g. atherton & r.c. sheppard, "solid phase peptide synthesis: practical methods ", 1989, IRL Press, Oxford.

For example, a resin with acid-labile linker groups is used 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. The amino-deprotection and coupling cycles are then repeated in alternating steps until the desired sequence is assembled.

Alternatively, the peptide V may be synthesized by solution phase peptide synthesis methods known in the art, in a stepwise manner from the carboxy terminus and/or by employing fragment condensation or ligation methods, using a global or minimal protection strategy. A combined liquid-solid phase fragment condensation method may also be employed.

Typically, the reactive side chain groups present (e.g., amino, hydroxyl, guanidino, and carboxyl groups) will be protected throughout the synthesis as described above. Many amino acid protecting groups are known to be available (see Grene, T.W. & Wuts, P.G.M (1991) for 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). Side chain protecting groups that may be employed include t-butyl (tBu), trityl (Trt), Boc and 2, 2, 5, 7, 8-pentamethylbenzodihydropyran-6-sulfonyl (Pmc). It will be appreciated that a wide range of other such groups are known in the art.

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

Conjugation of L to V:

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

Z is conjugated 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 may be used in the conjugation of Z and L. Particularly effective is the formation of amide bonds.

Example (b):

EXAMPLE 1 Ang II analogs with optical imaging dyes

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

Peptide analogues of Ang II were synthesized on an Applied Biosystems (Applied Biosystems)433A peptide synthesizer starting from 0.1mmol of Fmoc-Ile-Wang resin. An excess of 1mmol of preactivated amino acid (using O-benzotriazol-1-yl-N, N, N ', N' -tetramethyluronium Hexafluorophosphate (HBTU)) was used in the coupling step ending with arginine. The N-terminus was bromoacetylated with 0.5mmol of bromoacetic anhydride in DMF for 30 min. The bromoacetylated resin was then treated with a solution of 0.5mmol of N-Boc-ethylenediamine dissolved in N-methylpyrrolidone (NMP) for 30 minutes. The peptide was cleaved from the resin and the side chain protecting groups removed simultaneously by treatment in 5mL 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 resulting precipitated peptide was washed with ether and air dried.

23mg of crude peptide, 16mg of fluorescein NHS ester and 12. mu. L N-methylmorpholine (NMM) were dissolved in Dimethylformamide (DMF) and the reaction mixture was stirred overnight. The resulting reaction product was purified by preparative RP-HPLC (conditions: 5-50% B, where a ═ H, over 40 minutes₂O/0.1% TFA and B ═ CH₃CN/0.1% TFA; flow rate, 10 ml/min; column, Phenomenex Luna 5. mu.C 18(2) 250X 21.20mm) to give 19mg of pure fluorescein-peptide conjugate. The peptide was purified by analytical HPLC (conditions: gradient, 5-50% B in 10 min, where A ═ H₂O/0.1% TFA and B ═ CH₃CN/0.1% TFA; flow rate, 1 mL/min; column, Phenomenex Luna 3. mu.C 18(2) 50X 4.6 mm; detection, UV214 nm; product retention time 8.91 min). Further product identification was performed by electrospray mass spectrometry (MH)⁺Calculated value, 1355.6; MH⁺Found 1355.2).

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

EXAMPLE 2 tape for PET imaging¹⁸Ang II analogs of F

¹⁸F-(CH₂)₃-S-CH₂CO-NH-(CH₂)₂-Gly-Arg-Val-Tyr-Ile-His-Pro-Ile-OH

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

The peptide was cleaved from the resin and the side chain protecting groups removed simultaneously by treatment in 10mL 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 chloroacetylated peptide obtained was purified by preparative RP-HPLC. F¹⁸Conjugation of propyl thiol to purified chloroacetylated peptide to give peptides for PET imaging¹⁸F-labeled peptide.

Example 3^99mTc-labelled Ang II analogues

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

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

The peptide was cleaved from the resin and the side chain protecting groups removed simultaneously by treatment in 5mL 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.

By preparative RP-HPLC (0-30% B in 40 min, where A ═ H₂O/0.1% TFA and B ═ CH₃CN/0.1% TFA, on a Phenomenex Luna 5. mu.C 18(2) 250X 21.20mm column at a flow rate of 10mL/min) to give 12mg of pure chelate peptide conjugate. The product obtained is purified by analytical HPLC (conditions: gradient, 5-50% B in 10 min, where a ═ H₂O/0.1% TFA and B ═ CH₃CN/0.1% TFA; flow rate, 2 mL/min; column, Phenomenex Luna 3. mu.C 18(2) 50X 4.6 mm; detection, UV214 nm; product retention time 7.02 min). Further product identification was performed by using electrospray mass spectrometry (MH)⁺Calculated value, 1280.8; MH⁺Measured value, 1280.5)

Example 4 for^99mTc-labelled Ang II analogues

cPn216-CO-(CH₂)₃CO-NH-(CH₂)₂-Gly-Arg-Val-Tyr-Ile-His-Pro-Ile-OH

Ang II peptide analogs were synthesized on a peptide synthesizer using biosystems 433A, starting from 0.15mmol of Fmoc-Ile-Wang resin. An excess of 1mmol of preactivated amino acid (using HBTU) was used in the coupling step ending with arginine. The N-terminus was bromoacetylated with 0.75mmol of bromoacetic anhydride in DMF for 30 min. The resulting resin was then treated with a solution of 0.75mmol of N-Boc-ethylenediamine dissolved in NMP for 60 minutes. The peptide was cleaved from the resin and the side chain protecting groups removed simultaneously by treatment in 10mL 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.

5.5mg of peptide, 23mg of cPn216 tetrafluorothiophenyl esters (tetrafluoroothiophenylesters) and 10. mu.L of NMM were dissolved in DMF and the resulting reaction mixture was stirred for 3 hours. The product was obtained by preparative RP-HPLC (conditions: 0-30% B over 40 min, where a ═ H₂O/0.1% TFA and B ═ CH₃CN/0.1% TFA; flow rate, 10 mL/min; column, Phenomenex Luna5 μ C18(2)250 × 21.20mm) to yield 4.3mg of pure chelate-linker-peptide conjugate. The product was purified by analytical HPLC (conditions: gradient, 5-50% B10 min; where a ═ H₂O/0.1% TFA and B ═ CH₃CN/0.1% TFA; flow rate, 0.3 mL/min; column, Phenomenex Luna 3. mu.C 18(2) 50X 2 mm; detection, UV214 nm; product retention time 6.51 min). Further product identification was performed by using electrospray mass spectrometry (MH)⁺Calculated value, 1436.9; MH⁺Measured value, 1436.7)

Example 5 antiarrhythmic effects in porcine models 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 an artificial overpressure ventilation device. The heart was accessed via a midline sternotomy and the anterior pericardium was removed. The LAD artery is closed by ligation over the second branch and one of the branches is so closed. This leads to frequent extra ventricular contractions and is known from previous experiments with the same type of specimen to develop ventricular tachycardia and fibrillation within minutes. Within a few minutes after intravenous injection of 0.5mg of the compound of formula (X), the symptoms completely stabilized and sinus rhythm was restored. At this point it is noted that the mean blood pressure begins to rise. The extra-ventricular systole gradually reappears after about 15 minutes and a re-injection of 0.5mg of the compound of formula (X) is given. The symptoms again stabilize with a regular sinus rhythm. In the next hour, a stable sinus rhythm was recorded with a slightly reduced but substantially constant blood pressure. The pigs were then given a lethal amount of potassium chloride to terminate the trial. Analysis of ECG recordings before and after the first injection of compound (X) indicated a reduction in duration of the QRS complex, suggesting that the antiarrhythmic effect may be associated with an increase in myocardial conduction rate.

A similar set of observations was made in the second pig trial.

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

Example 6N- ((CH₂)₆Synthesis of-tetramine) -Gly-Arg-Val-Tyr-Ile-His-Pro-Ile-OH

Starting from 0.25mmol of Fmoc-Ile-Wang resin, an Fmoc/tert-butyl strategy is adopted to synthesize an Arg-Val-Tyr-Ile-His-Pro-Ile peptide sequence on an applied biosystem 433A peptide synthesizer. An excess of 1mmol 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-terminus was bromoacetylated with 1mmol of bromoacetic anhydride in dimethylformamide for 30 minutes. The resulting bromoacetylated resin (0.05mmol) was then treated with 0.15mmol of tetra-Boc-tetraamine chelate solution dissolved in dimethylformamide for 60 minutes.

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

The 25mg crude product was purified by preparative RP-HPLC (0-30% B over 40 min, where a ═ H₂O/0.1% TFA and B ═ CH₃CN/0.1% TFA on a Phenomenex Luna5 μ C18(2) 250X 21.20mm column at a flow rate of 10mL/min) to give 6.5mg of semi-purified product. Second purification step of semi-purified product (a ═ H)₂O/0.1% HCOOH andB＝CH₃CN/0.1% HCOOH, the remainder being under the same conditions as above) to give 3mg of a pure product. The product was purified by analytical HPLC (conditions: gradient, 0-30% B in 10 min, where A ═ H₂O/0.1% HCOOH and B ═ CH₃CN/0.1% HCOOH; flow rate, 0.3 mL/min; column, Phenomenex Luna 3. mu.C 18(2) 50X 2 mm; detection, UV214 nm; product retention time, 5.34 min). Further product identification was performed using electrospray mass spectrometry (MH)⁺Calculated value, 1196.8; MH⁺Measured value, 1196.7)

Example 7 Hcy 3-5; synthesis of N-cPn216-Gly-Arg-Hcy-Tyr-Hcy-His-Pro-Ile-OH

Starting from 0.25mmol of Fmoc-Ile-Wang resin, an Fmoc/tert-butyl strategy was employed to synthesize an Arg-Hcy-Tyr-Hcy-His-Pro-Ile peptide sequence on an applied biosystems 433A peptide synthesizer. An excess of 1mmol 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-terminus was bromoacetylated with 1mmol of bromoacetic anhydride in dimethylformamide for 30 minutes. The resulting bromoacetylated resin (0.05mmol) was then dissolved in dimethylformamide with 0.25mmol of cPn216 (0.05mmol)^99mTc-chelating agents¹)(¹See patent WO20030049 for details of synthesis) solution treatment for 60 minutes.

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

The crude linear product was purified by preparative RP-HPLC (5-50% B40 min, where a ═ H₂O/0.1% TFA and B ═ CH₃CN/0.1% TFA, pure in a Phenomenex Luna 5. mu.C 18(2) 250X 21.20mm column at a flow rate of 10mL/min)To obtain 20mg of pure linear product. The linear product was dissolved in 2mL DMSO and 200mL water, and the resulting solution was adjusted to pH8 with aqueous ammonia. The resulting solution was stirred for 26 hours and then adjusted to pH2 with trifluoroacetic acid.

The resulting cyclic product was purified by preparative RP-HPLC (0-30% B, remainder under the same conditions as above) to give 12mg of pure cyclic product. The product obtained is purified by analytical HPLC (conditions: gradient, 5-50% B10 min, where A ═ H₂O/0.1% TFA and B ═ CH₃CN/0.1% TFA; flow rate, 0.3 mL/min; column, Phenomenex Luna 3 μ C18(2)50 × 2mm column; detection, UV214 nm; product retention time, 6.01min) were analyzed. Further product identification was achieved by using electrospray mass spectrometry (MH)⁺Calculated value, 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

<210>1

<211>8

<212>PRT

<213> Artificial sequence

<220>

<223> synthetic peptide

<220>

<221>misc_feature

<222>(1)..(1)

<223> any amino acid

<220>

<221>misc_feature

<222>(2)..(2)

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

<220>

<221>misc_feature

<222>(8)..(8)

<223> hydrophobic side chain-containing amino acid

<400>1

Xaa Xaa Val Tyr Ile His Pro Xaa

1 5

<210>2

<211>8

<212>PRT

<213> Artificial sequence

<220>

<223> synthetic angiotensin II

<400>2

Asp Arg Val Tyr Ile His Pro Phe

1 5

<210>3

<211>8

<212>PRT

<213> Artificial sequence

<220>

<223> synthetic peptide

<400>3

Gly Arg Val Tyr Ile His Pro Ile

1 5

<210>4

<211>8

<212>PRT

<213> Artificial sequence

<220>

<223> synthetic peptide

<220>

<221>MISC_FEATURE

<222>(2)..(2)

<223> methylated Arg

<220>

<221>MISC_FEATURE

<222>(8)..(8)

<223>D-Phe

<400>4

Gly Xaa Val Tyr Ile His Pro Xaa

1 5

<210>5

<211>8

<212>PRT

<213> Artificial sequence

<220>

<223> synthetic peptide

<220>

<221>MISC_FEATURE

<222>(8)..(8)

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

<400>5

Gly Arg Val Tyr Ile His Pro Xaa

1 5

Claims (11)

1. A medicine, characterized by formula (I)

Z-(L)_n-V (I)

Wherein,

v represents a peptide having the binding sequence-X¹-X²-Val-Tyr-Ile-His-Pro-X³The peptide of (a) above (b),

l represents an optional linker, and L represents an optional linker,

z represents a group optionally carrying an imaging moiety M,

n is a number of 0 or 1,

X¹represents an amino groupThe acid is added to the mixture of the acid,

X²represents Arg or N-alkylated Arg or Arg mimetic,

X³refers to an amino acid comprising a hydrophobic side chain,

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

z is optionally joined to amino acid X via a linker L¹Form a bond, and

the presence of M represents an imageable moiety that can be detected directly or indirectly in a diagnostic imaging procedure.

2. The medicament of claim 1, which is effective in treating heart failure, arrhythmia and other diseases in which fibrosis is prominent and in treating COPD, liver fibrosis and arteriosclerosis.

3. The medicament of claim 1, wherein M is an in vivo imageable moiety for use in diagnosis.

4. A medicament as claimed in any one of claims 1 to 3, wherein Z represents a chelating agent of formula (VII)

Wherein:

each R¹、R²、R³And R⁴Independently is H or C_1-10Alkyl radical, C_3-10Alkylaryl group, C_2-10Alkoxyalkyl group, C_1-10Hydroxyalkyl radical, C_1-10Alkylamine, C_1-10Fluoroalkyl, or 2 or more R groups together with the atoms linking them form a saturated or unsaturated carbocyclic or heterocyclic ring.

5. A medicament as claimed in any one of claims 1 to 4, wherein Z represents a chelator of formula (XI)

Wherein Q₁-Q₆Independently is a Q group, wherein Q is H, alkyl, aryl, or an amine protecting group,

W₁is-NR-, -CO₂a-CO-, -NR (C ═ S) -, -NR (C ═ O) -, -CONR-or Q group;

each Y is independently a D-or L-amino acid, -CH₂-、-CH₂OCH₂-or-OCH₂CH₂An O-or X group;

p is an integer having a value of 1 to 8;

q is an integer having a value of 0 to 30;

r is H, C_1-4Alkyl radical, C_2-4Alkoxyalkyl group, C_1-4Hydroxyalkyl or C_1-4A fluoroalkyl group;

q is

A is a counter ion.

6. A medicament as claimed in any one of claims 1and 3 to 5, wherein M represents a gamma-emitting moiety for Radio or SPECT imaging, comprising⁶⁷Ga、¹¹¹In、¹²³I、¹²⁵I、¹³¹I、^81mKr、⁹⁹Mo、^99mTc、²⁰¹Tl and¹³³Xe。

7. the medicament as claimed in the preceding claim, for use in therapy, having formula (X) or formula (XII)

Formula (X)

Formula (XII)

Or as a diagnostic agent having formula (Xa) or (XIIa)

8. A pharmaceutical formulation comprising a compound of formula (I) as claimed in claim 1 together with one or more pharmaceutically acceptable additives and/or excipients.

9. Use of a medicament according to claim 1 for the treatment and/or diagnosis of heart failure, cardiac arrhythmias and other diseases where fibrosis is prominent, in particular COPD, liver fibrosis and arteriosclerosis.

10. An in vivo diagnostic method for heart failure and other diseases where fibrosis is prominent, in particular COPD, liver fibrosis and arteriosclerosis, in a subject, comprising administering a medicament of formula (I) as claimed in claim 1and subsequently imaging said subject locally or systemically.

11. A kit for preparing a radiopharmaceutical composition of formula (I), which comprises a peptide-chelate conjugate and a reducing agent.

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