DE69735006T2 - CONTRAST AGENTS - Google Patents

Description

These This invention relates to diagnostic imaging techniques, in a disease state using a targeted contrast agent can be imaged and targeted at contrast agents that are used to Use in such techniques are suitable. In particular, refers the invention relates to the use of such contrast agents, where the targeting vector binds to angiotensin II receptors.

Angiotensin II (hereafter referred to as A II) is an octapeptide (Asp-Arg-Val-Tyr-Ile-His-Pro-Phe) which is the primary active species in the renin-angiotensin-aldosterone system (the RAS system). is that regulates the blood pressure and the electrolyte and water balance. The RAS system is schematic in 1 shown which on 1 in the article by Foote et al. in Ann. Pharmacother. 27: 1495-1503 (1993).

A II exercises its biological effects over Interaction with specific receptors found in many tissues present in the human or animal body (e.g., blood vessels, heart, Brain, liver, kidney, uterus and ovaries). While it's one of its main effects is to promote vasoconstriction, A II has several other effects that lead to increased blood pressure and sodium retention. For example, in response to stimulation of A II receptors Aldosterone released in the adrenal cortex and this stimulated the sodium and water retention and potassium excretion in the distal tubules and cortical collecting ductus of the kidney. A II owns also a direct effect on the kidney, including glomerular hypertrophy and increased proximal tubule sodium reabsorption. It also works A II central to stimulate thirst and the anti-diuretic hormone release improve, resulting in an increased intravascular volume leads.

Accordingly, became the suppression of A II effects is used therapeutically, e.g. at the Management of hypertension and congestive heart failure. This has been achieved in a number of ways: through use of renin inhibitors that increase the conversion of angiotensinogen Block angiotensin I (the precursor of A II); through the use of Angiotensin Converting Enzyme Inhibitors (ACE) Inhibitors Conversion of Angiotension I to A II block (and also bioconversion blocked by bradykinin and prostaglandins); through the use of anti-A II antibodies; and by the use of A II receptor antagonists.

It has been found that different types of A II receptors (binding sites) exist within the body and that the A II binding has different effects at different binding sites. Thus, the AT1 receptor mediates the major cardiovascular effect of RAS and is inhibited by the A II receptor antagonists losartan and DTT, while the other major receptor family, AT2, is inhibited by PD 123177 and its structural analogues. Other receptors for A II other than AT1 and AT2 are known and generally referred to as AT _atypical (see Kang et al., Am., Heart J. 127: 1388-1401 (1994)).

U.S. Patent No. 5,444,069 discloses the use of 4,5,6,7-tetrahydro-1H-imidazo [4,5-c] pyridine-6-carboxylic acids and analogs thereof in antagonizing the binding of angiotensin II to AT ₂ receptors to treat memory loss.

It has now been found that it is possible to image A II receptor sites in vivo using targeted contrast agents in which the targeting vector has an affinity for A II receptor sites. The A II receptors are generally located within the cardiovascular system and accessible to such contrast agents when administered into the bloodstream. Accordingly, using such targeted contrast agents, it is possible to detect diseases and disorders such as cardiac insufficiency, atherosclerosis and limited blood flow, as well as other vascular diseases and disorders and also to monitor the progress of treatment for such diseases and disorders. From one aspect, therefore, the invention provides a composition of matter of formula I. VLR (I) wherein V is an organic group having a binding affinity for an angiotensin II receptor site, L is a linker moiety or a bond, and R is a moiety detectable in in vivo imaging of a human or animal body, provided that V for angiotensin or a peptidic angiotensin derivative or analog, VLR is other than a non-metallic radionuclide-substituted peptide (eg ¹²⁵ I-substituted angiotensin II) and LV is other than simply a peptide having a chelator amide linked to a side chain thereof.

Prefers, where R is a radionuclide, it is an iodine, directly or indirectly is bound to V, or it is a metal ion that is chelated by a Group is chelated in L and / or it is separable from V by biodegradation an organic linker group L. (It should be noted that Hydrogen radionuclides while they are detectable in vitro do not meet the requirement that R for in vivo imaging is detectable.)

Prefers is where V for Angiotensin stands, R not a metal ion.

In many cases the substance composition of the formula I becomes a characterizable Be connected. In others, it can be a combination of compounds that are interconnected or otherwise associated, e.g. are conjugated. For the sake of simplicity, the composition of matter hereinafter referred to as a means.

From In another aspect, the invention provides a pharmaceutical A composition comprising an effective amount (e.g. Amount to reinforce the image contrast is effective in in vivo imaging) By means of the formula I together with at least one pharmaceutical effective carrier or excipient.

From In yet another aspect, the invention provides Use of an agent of formula I for the preparation of a contrast medium ready for use in a diagnostic procedure that involves the administration of the contrast medium to a living subject and the generation of an image of at least a part of the subject.

From In still another aspect, the invention ceases Method for generating an image of a living human or non-human (preferably mammal or bird) animal Subject ready, which involves administering a contrast agent to the subject and generating an image of at least a part the subject to whom the contrast agent has been distributed, e.g. by X-ray, MR, Scintigraphy, PET, SPECT, electrical impedance, light or magnetometric Imaging modalities, characterized in that as the contrast agent is a means of Formula I is used.

From In another aspect, the invention provides a method to monitor the effect of treating a human or non-human animal Subject with a drug ready to fight or effects which are associated with angiotensin II, wherein the Method involves administering to the subject a means of Formula I and detecting the uptake of the agent by angiotensin II receptors, wherein administration and detection are optional, but preferably repeatedly, e.g. before, during and after treatment with the drug.

From In still another aspect, the invention provides a Process for the preparation of an agent of formula I, wherein the process comprises conjugating ((i) an organic compound with a binding affinity for one A II receptor on (ii) a compound used in a diagnostic imaging procedure is detectable or a chelating compound, and, if necessary, Metalating the chelating groups in the resulting conjugate with a metal ion in a diagnostic imaging procedure is detectable.

The Agents of formula I have three characteristic components: a vector (V); a linker (L); and a reporter (R). Of the Vector must have the ability to target the compound on A II receptors, the reporter must be detectable in a diagnostic in vivo imaging procedure be; and the linker has to couple the vector to the reporter, at least until the imaging procedure has been completed.

vectors

When the vector may be any compound having an affinity for A II receptors. This may be a peptide, such as A II itself, or an analog, such as saralasin (Sar-Arg-Val-Tyr-Val-His-Pro-β-Ala) or a non-peptide, such as iosartan or PD-123177, although non-peptides are preferred. On similar Way, a vector can be a compound (like losartan) that has one pronounced affinity for one Type of A II receptor as for has other types, or a compound (such as A II itself), the a general affinity for all A Has II receptors. Compounds with a stronger affinity for certain Types of A II receptors (such as AT1 or AT2) are generally to be favoured.

Prefers the agent is a compound that is not unacceptable biological Reaction triggers, in particular compounds which act as A II receptor antagonists, and not trigger the reactions associated with A II itself, in particular the blood pressure-modifying reactions. However, if desired, the biological Reactions modified by administration of a therapeutic agent be, e.g. before, at the same time or after the administration of the By means of the formula I.

Under can peptidic vectors Oligopeptide with the motif Arg-Val-Tyr-Ile-His-Pro or an analogous motif in which one of the five Amino acid residues either individually or together with structural or functional Isosterene can be varied so that the structural or functional Motif is conserved so as to interact effectively with Allow angiotensin II receptor sites. As an example, the Motifs Arg-Val-Tyr * -XX-His-Pro (where Tyr * is an optionally modified Tyr, e.g. Me-Tyr and XX is an amino acid, e.g. Ile or Val) preferred, e.g. A II and especially Saralesin and Sarmesin. However, where A II or such near angiotensin analogues are used as the vector it can be desired be, linker-reporter combinations others, other than metallated chelating agents (e.g., including a radionuclide or a paramagnetic metal ion) directly attached to the peptidic Structure are bound; so it may be desirable to use a non-metallic To use radionuclide reporters or a particulate reporter to use.

Other suitable oligopeptide vectors can be easily determined using a phage display, combinatorial Chemistry (both peptidic and non-peptidic combinatorial chemistry), HTS (high throughput synthesis) and CAM.D techniques.

Under nonpeptidic vectors are losartan and PD 123177 is preferred. Other suitable non-peptidic vectors include heterocyclic Compounds (such as imidazoles, condensed imidazoles, xanthines and Pyridones). Examples of suitable non-peptide vectors are WO-91/17148, US-A-4355040, WO-91/18888, WO-91/19715, WO-91/15209, EP-A-420237, EP-A-459136 and US-A-5338744. The development of suitable non-peptide A II receptor antagonists are discussed e.g. from Timmermans et al., In TiPS 12: 55-62 (1991) and Pharm. Rev. 45: 20-251 (1993) and van Meel et al. in Arzneimittel-Forsch./Drug Res. 43: 242-246 (1993).

Suitable non-peptidic vectors will generally contain a 5- or 6-membered unsaturated heterocyclic N ₁ , N ₂ or N ₃ ring optionally bearing a fused 5- or 6-membered unsaturated, optionally heterocyclic ring and optionally pendant Bears groups which comprise one or more (eg 1, 2 or 3) 5- or 6-membered homo- or heterocyclic aryl groups, eg aryl-methyl-biphenyltetrazoles. Examples include DuP 753, L-158809, SR-47436, GR 117289, SKF 108566, BIBS 39, BIBS 222, ExP 3892 and CG 48933.

wobei R₁ und R₂ für Carboxy-Gruppen oder C_1-6-Alkyl-Gruppen stehen, die gegebenenfalls mit Oxa-, Oxo-, Phenyl-, Amino-, Carboxyl- oder Hydroxy-Gruppen substituiert sind, oder R₁ zusätzlich eine Oxo-Gruppe repräsentieren kann, und
R₃ und R₄ für Methyl-biphenyl-carboxyl- oder Methyl-biphenyl-tetrazol-Gruppen oder C_1-6-Alkyl-Gruppen stehen, die gegebenenfalls mit Oxa-, Oxo-, Phenyl-, Amino-, Carboxyl- oder Hydroxy-Gruppen substituiert sind, wobei R₄ nur vorhanden ist, wenn R₁ für Oxo steht, wobei bevorzugt eine der Gruppen R₃ und R₄ eine Biphenyl-enthaltende Gruppe repräsentiert und eine der Gruppen R₁, R₂, R₃ und R₄ eine Alkyl-Gruppe repräsentiert, die mit Oxa-, Oxo-, Phenyl-, Amino-, Carboxyl- oder Hydroxy-Gruppen substituiert ist.Therefore, the vector compound may be, for example, a compound of formula II

wherein R ₁ and R _{2 are} carboxy groups or C _1-6 alkyl groups which are optionally substituted by oxo, oxo, phenyl, amino, carboxyl or hydroxy groups, or R ₁ additionally one Oxo group can represent, and
R ₃ and R _{4 are} methyl-biphenyl-carboxyl or methyl-biphenyl-tetrazole groups or C _1-6 -alkyl groups, optionally with oxa, oxo, phenyl, amino, carboxyl or hydroxy Groups are substituted, wherein R ₄ is present only when R _{1 is} oxo, wherein preferably one of the groups R ₃ and R _{4 represents} a biphenyl-containing group and one of the groups R ₁ , R ₂ , R ₃ and R ₄ represents an alkyl group substituted with oxo, oxo, phenyl, amino, carboxyl or hydroxy groups.

In such compounds the amino, hydroxy, carbonyl or carboxy functions are readily used, to conjugate the vector to the reporter.

wobei R₃₀ steht für

R₃₁ steht für COOH, Tetrazol-5-yl, NH₂SO₂CF₃, SO₂NHCO-Phenyl, SO₂NHCO-Cyclopropyl und R₁₀ und R₂₀ stehen für

wobei R₄₀ steht für

R₃₁ wie oben definiert ist und R₂₀ und R₃₉ stehen fürThus, examples include such triazole vectors

where R ₃₀ is for

R ₃₁ is COOH, tetrazol-5-yl, NH ₂ SO ₂ CF ₃ , SO ₂ NHCO-phenyl, SO ₂ NHCO-cyclopropyl and R ₁₀ and R ₂₀ are

where R ₄₀ is for

R _{31 is} as defined above and R ₂₀ and R ₃₉ stand for

wobei die Doppelbindung, die mit einem Sternchen markiert ist, gegebenenfalls durch eine Einfachbindung ersetzt ist; R₁₀₀ für ein Wasserstoffatom, eine Phenyl-Gruppe, die gegebenenfalls mit mindestens einer Hydroxy- oder Amino-Gruppe substituiert ist, oder eine C_1-6-Alkyl-Gruppe, die gegebenenfalls mit einer Hydroxy-Gruppe substituiert ist oder gegebenenfalls über einen Sauerstoff, Schwefel oder Stickstoff gebunden ist, steht; R₄₀₀ für eine Arylmethyl-Gruppe steht, z.B. eine Methylbiphenyl (gegebenenfalls substituiert auf einem Phenylring durch eine Säurefunk tion, z.B. ein Carboxyl oder ein Ester davon, z.B. eine (C_1-6-Alkoxy)-carbonylgruppe) oder Methylbiphenyltetrazol-Gruppe, die über die Methylgruppe gebunden ist, oder eine gegebenenfalls C_1-6-Alkyl-, Amino- und/oder Hydroxy-substituierte Benzylgruppe; R₃₀₀ für eine Carboxymethyl-Gruppe oder eine gegebenenfalls Hydroxysubstituierte C_1-6-Alkyl-Gruppe steht, z.B. eine Hydroxy-Methylgruppe, und R₂₀₀ für Chlor steht oder R₃₀₀ und R₂₀₀ zusammen mit den zwischengeschalteten Kohlenstoffen einen kondensierten 6-gliedrigen carbocyclischen oder heterocyclischen aromatischen Ring (enthaltend 0, 1 oder 2 Ring-Stickstoffe) bilden, der gegebenenfalls an einem Ring-Kohlenstoff mit einer Hydroxy-, Carboxyl-, C_1-6-Alkyl- oder Acyloxy-Gruppe (wobei die Acyl-Gruppe eine C_1-6-Alkyl-, Phenyl- oder Benzyl-Gruppe umfasst) oder an einem Ring-Stickstoff mit einer Acyl-Gruppe substituiert ist (z.B. eine C_1-6-Alkylcarbonyl-Gruppe, die gegebenenfalls mit Phenyl-Gruppen substituiert ist). Bevorzugt enthält mindestens eine von R₁₀₀, R₂₀₀, R₃₀₀, und R₄₀₀ eine Hydroxy-, Amino-, Thiol-, Mercapto-, Aldehyd-, Keton-, oder Carbonsäure-Gruppe, oder eine Vielfachbindung (z.B. eine C=C-Bindung) oder eine beliebige andere funktionelle Gruppe, die zum Binden des Vektors an die Reportereinheit verwendet werden kann.Similarly, the vector may be a compound of Formula III

wherein the double bond marked with an asterisk is optionally replaced by a single bond; R _{100 represents} a hydrogen atom, a phenyl group optionally substituted with at least one hydroxy or amino group, or a C _1-6 alkyl group optionally substituted with a hydroxy group or optionally an oxygen , Sulfur or nitrogen is bound; R _{400 is} an arylmethyl group, for example a methylbiphenyl (optionally substituted on a phenyl ring by an acid function, eg a carboxyl or an ester thereof, eg a (C _1-6 -alkoxy) -carbonyl group) or methylbiphenyltetrazole group bonded via the methyl group, or an optionally C _1-6 alkyl, amino and / or hydroxy-substituted benzyl group; R _{300 is} a carboxymethyl group or an optionally hydroxy-substituted C _1-6 alkyl group, for example a hydroxy methyl group, and R _{200 is} chlorine or R ₃₀₀ and R ₂₀₀ together with the interposed carbons is a condensed 6-membered carbocyclic or heterocyclic aromatic ring (containing 0, 1 or 2 ring nitrogens) optionally substituted on a ring carbon having a hydroxy, carboxyl, C _1-6 alkyl or acyloxy group (wherein the acyl group is a C _1-6 alkyl, phenyl or benzyl group) or substituted on a ring nitrogen with an acyl group (eg a C _1-6 alkylcarbonyl group optionally substituted with phenyl groups) , Preferably, at least one of R ₁₀₀ , R ₂₀₀ , R ₃₀₀ , and R _{400 contains} a hydroxy, amino, thiol, mercapto, aldehyde, ketone, or carboxylic acid group, or a multiple bond (eg, a C = C Bond) or any other functional group that can be used to attach the vector to the reporter moiety.

sein, wobei R₃₀₉ für Hydroxymethyl oder Carboxy steht, oder von der Formel

sein, wobei R₁₀₁-Phenyl- für 3,4-Dihydroxyphenyl, 4-Aminophenyl oder 2-Hydroxyphenyl steht, oder von der Formel

sein, wobei R₃₀ wie oben definiert ist, z.B. Methyl-Biphenyl-Tetrazol oder Methylbiphenylcarboxyl, und R₂₀₂ und R₃₀₂ sind wie folgtThus, for example, the compound of formula III of the formula

where R _{309 is} hydroxymethyl or carboxy, or of the formula

wherein R _{101 is} phenyl for 3,4-dihydroxyphenyl, 4-aminophenyl or 2-hydroxyphenyl, or of the formula

wherein R _{30 is} as defined above, for example, methyl-biphenyl-tetrazole or methylbiphenylcarboxyl, and R ₂₀₂ and R ₃₀₂ are as follows

sein, wobei R₃₀ wie oben definiert ist, oder von der Formel

sein, wobei R₃₀ wie oben definiert ist und R₁₀₂ für C_1-3-Alkyl, gebunden über O, S oder NH, steht, oder von der Formel

sein, wobei R₇₇ für Hydroxy oder Amino steht und die gestrichelte Bindung eine Doppel- oder eine Einfachbindung ist, bevorzugt eine Doppelbindung, wo R₇₇ für Amino steht, und eine Einfachbindung, wo R₇₇ für Hydroxy steht.Alternatively, the vector of formula III may be of the formula

wherein R _{30 is} as defined above or of the formula

wherein R _{30 is} as defined above and R _{102 is} C _1-3 alkyl attached via O, S or NH, or of the formula

where R _{77 is} hydroxy or amino and the dotted bond is a double or a single bine is preferably a double bond where R _{77 is} amino and a single bond where R _{77 is} hydroxy.

wobei R₅₀₁ für eine gegebenenfalls fluorierte C_1-6-Alkyl-Gruppe steht, R₅₀₄ für eine gegebenenfalls fluorierte C_1-6-Alkyl-Gruppe steht, R₅₀₂ für eine Carboxy-, Hydroxy-, C_1-6-Alkyl- oder CHO-Gruppe steht; oder die Doppelbindung, die mit einem Sternchen markiert ist, durch eine Einfachbindung ersetzt ist und R₅₀₄ für eine =O oder =NR₅₀₅-Gruppe steht (wobei R₅₀₅ für C_1-6-Alkyl steht), R₅₀₂ eine gegebenenfalls fluorierte C_1-6-Alkylgruppe steht und R₅₀₃ für eine Carboxyphenyl-Gruppe steht; und R₃₀ wie oben definiert ist (z.B. wenn R₃₁ für Tetrazol-5-yl, NH₂SO₂CF₃, SO₂NHCO-Cyclopropyl oder SO₂NHCO-Phenyl).In addition, the vector may be a compound of formula IV

where R _{501 is} an optionally fluorinated C _1-6 -alkyl group, R _{504 is} an optionally fluorinated C _1-6 -alkyl group, R _{502 is} a carboxy, hydroxy, C _1-6 -alkyl group or CHO group; or the double bond marked with an asterisk is replaced by a single bond and R _{504 is} an = O or = NR ₅₀₅ group (wherein R _{505 is} C _1-6 alkyl), R _{502 is} an optionally fluorinated C _1-6 alkyl group and R _{503 represents} a carboxyphenyl group; and R _{30 is} as defined above (eg when R _{31 is} tetrazol-5-yl, NH ₂ SO ₂ CF ₃ , SO ₂ NHCO-cyclopropyl or SO ₂ NHCO-phenyl).

wobei R₂₇ für Methyl oder C₂F₅ steht,
R₂₆ für COOH, CHO oder CH₂OH steht,
R₂₅ für Butyl oder Propyl steht,
R₂₈ für =O oder =NCH₃ steht, und
R₂₁ für Tetrazol-5-yl, NH₂SO₂CF₃, SO₂NHCO-Cyclopropyl oder SO₂NHCO-Phenyl steht.For example, the vector of formula IV may be of the formula

where R _{27 is} methyl or C ₂ F ₅ ,
R _{26 is} COOH, CHO or CH ₂ OH,
R _{25 is} butyl or propyl,
R _{28 is} = O or = NCH ₃ , and
R _{21 is} tetrazol-5-yl, NH ₂ SO ₂ CF ₃ , SO ₂ NHCO-cyclopropyl or SO ₂ NHCO-phenyl.

Other Compounds suitable for use as vectors include Compounds such as those described in WO 92/18529, US-A-5156977, EP-A-435827, EP-A-424317, EP-A-420237, EP-A-426021, WO 92/09556, WO 91/19715, WO 91/18888, WO 91/17148, EP-A-446062, WO 92/09563, WO 91/15479, WO 91/15209, EP-A-459136, EP-A-445811, EP-A-442473, EP-A-430300, US-A-4355040, US-A-4340598 and US-A-5338744.

The Vectors of the formulas II to IV can produced by the techniques described in the patent publications are described, which is incorporated herein by reference.

CAM-D and other possible ones Identification and evaluation techniques as mentioned above can also used to possible to further evaluate non-peptidic vectors.

Thus, it is also possible to obtain molecules that specifically bind to angiotensin II receptors by directly screening molecular libraries. For example, phage libraries displaying small peptides could be used for such selection. The selection can be made by simply mixing the receptors and the phage display library and eluting the phages that bind to the receptors. If desired, selection may be made under "physiological conditions" (eg, in the blood) to eliminate peptides that cross-react with blood components. Binding moieties identified in this manner may be isolated (by chemical conjugation or via peptide synthesis, or at the DNA level for recombinant vectors) to a linker molecule, which is a generic one Forming tool for binding any vector molecules to the reporter.

Vector molecules can also are generated from combinatorial libraries without necessarily to know the exact molecular target by functional selection (in In vitro, ex vivo or in vivo) of molecules to be imaged Tie region / structure.

The Vector unit in the agents of formula I is preferably an educational constant for R-L-V possess: angiotensin II receptor site conjugation of at least 100th

As mentioned above, the agents of formula I include vector, linker, and reporter moieties. A linker moiety can serve to bind a vector to a reporter; alternatively, it may connect more than one vector and / or more than one reporter. In the same way, a reporter or vector may be linked to more than one linker. Using a plurality of reporters in this manner (eg, multiple linker reporter moieties attached to one or more reporters bound to a reporter that is itself bound to a vector) may allow the detectability of the Contrast agent is increased (eg by increasing its radiopacity, echogenicity or relaxivity), or may allow it to be detected in more than one imaging modality. Use of a plurality of vectors in this manner may increase the targeting efficiency of the contrast agent or may enable the contrast agent to target more than one site, eg, different reporters for an agent that has receptor heterogeneity. Thus, for example, the agent of formula I may contain vector units having affinity sites other than angiotensin receptors, eg having affinities for cell surfaces on body-duct wall surfaces. Accordingly, the agent may contain vectors such as antibody fragments and oligopeptides, eg, containing RGD or cell surface analog binding peptide motifs (eg, as described in EP-A-422937 and EP-A-422938 (Merck)) or other vectors as described in U.S. Pat GB 9700699.3 , Such additional vectors may also be selected from any of the molecules that are naturally concentrated in vivo in a selected target organ, tissue, cell or cell groups, or elsewhere in a mammalian body. These may include amino acids, oligopeptides (eg hexapeptides), molecular recognition units (MRU's), single chain antibodies (SCA's), proteins, non-peptidic organic molecules, Fab fragments and antibodies. Examples of targeted molecules include polysaccharides (eg CCK and hexapeptides), proteins (such as lectins, asialofetuin, polyclonal IgG, blood clotting proteins (eg hirudin), lipoproteins and glycoproteins), hormones, growth factors, clotting factors (such as PF4), polymerized fibrin fragments (eg, E ₁ ), Serum amyloid precursor (SAP) proteins, low density lipoprotein (LDL) precursors, serum albumin, surface proteins intact red blood cells, receptor binding molecules such as estrogens, liver specific proteins / polymers such as galactosyl neoglycoalbumin (NGA) ( see Vera et al in Radiology 151: 191 (1984)) N- (2-hydroxypropyl) methacrylamide (HMPA) copolymers with varying numbers of bound galactosamines (see Duncan et al., Biochim. Biophys. Acta 880: 62 (1986)), and allyl and 6-aminohexyl glycosides (see Wong et al., Carbo. Res. 170: 27 (1987)) and fibrinogen. The targeted protein may also be an antibody. The choice of antibody, particularly the antigenic specificity of the antibody, will depend on the particular, intended target site for the agent. Monoclonal antibodies are preferred over polyclonal antibodies. Production of antibodies that react with a desired antigen is well known. Antibody preparations are commercially available from a variety of sources. The fibrin fragment E ₁ can be prepared as described by Olexa et al. in J. Biol. Chem. 254: 4925 (1979). The production of LDL precursors and SAP proteins is described by de Beer et al. in J. Immunol. Methods 50:17 (1982).

It It is especially preferred that such extra vectors bind in this way should slow down the movement of the agent in the bloodstream, but is not prevented and anchor it in its place, though it is bound to an A II receptor site.

left

The Linker component of the contrast agent is most easily a bond between the vector and reporter units. More general is the Left, however, a mono- or to provide a multimolecular skeleton that is covalent or non-covalent binding one or more vectors to one or more receptors, e.g. a linear, cyclic, branched or reticular molecular Skeleton or a molecular aggregate, with built-in or attached Groups which are covalently or non-covalently, e.g. coordinatively, with the vector and reporter units are connected, or the like Encapsulate, capture or anchor units.

So can bind a reporter unit to a desired one Vector can be achieved by covalent or non-covalent means, usually containing an interaction with one or more functional Groups positioned on the reporter and / or vector. Examples of chemically reactive functional groups responsible for this Purpose can be used include amino, hydroxyl, sulfhydryl, carboxyl and carbonyl groups, as well as hydrocarbon groups, vicinal diols, thioethers, 2-amino alcohols, 2-aminothiols, guanidinyl, imidazolyl and phenol groups.

Covalent coupling of reporter and vector can therefore be effected using linking agents containing reactive moieties capable of reacting with such functional groups. Examples of reactive moieties capable of reacting with sulfhydryl groups include alpha-haloacetyl compounds of the type X-CH ₂ CO- (where X = Br, Cl or I) which exhibit particular reactivity for sulfhydryl groups but can also be used to modify imidazolyl, thioether, phenol and amino groups as described by Gurd, FRN in Methods Enzymol. (1967) 11, 532. N-maleimide derivatives are also considered selective to sulfhydryl groups, but may additionally be useful in coupling to amino groups under certain conditions. Reagents such as 2-iminothiolane, eg as described by Traut, R. et al. in Biochemistry (1973) 12, 3266, which introduce a thiol group by conversion of an amino group, may be considered as sulfhydryl reagents if binding occurs through the formation of disulfide bridges. Thus, reagents that introduce reactive disulfide bonds into either the reporter or the vector may be useful, as binding can be induced by disulfide exchange between the vector and the reporter; Examples of such reagents include Ellman's reagent (DTNB), 4,4'-dithiopyridine, methyl 3-nitro-2-pyridyl disulfide and methyl 2-pyridyl disulfide (described by Kimura, T. et al., In Analyt. Biochem ) 122, 271).

• i) α-Haloacetyl-Verbindungen, die eine Spezifität gegenüber Amino-Gruppen bei nicht Vorhandensein reaktiver Thiol-Gruppen zeigen und vom Typ X-CH₂CO- sind (wobei X = Cl, Br oder I), z.B. wie beschrieben von Wong, Y-H. H. in Biochemistry (1979) 24, 5337;
• ii) N-Maleimid-Derivate, die mit Amino-Gruppen reagieren können, entweder über eine Reaktion des Michael-Typs oder durch Acylierung durch Addition an die Ring-Carbonyl-Gruppe, wie beschrieben von Smyth, D. G. et al. in J. Am. Chem. Soc. (1960) 82, 4600 und Biochem. J. (1964) 91, 589;
• iii) Arylhalogenide, wie reaktive nitrohalo-aromatische Verbindungen;
• iv) Alkylhalogenide, wie beschrieben von McKenzie, J. A. et al. in J. Protein Chem. (1988) 7, 581;
• v) Aldehyde und Ketone, die zur Schiff-Basen-Bildung mit Amino-Gruppen fähig sind, wobei die gebildeten Addukte üblicherweise durch Reduktion stabilisiert werden, um ein stabiles Amin zu ergeben;
• vi) Epoxid-Derivate, wie Epichlorhydrin und Bisoxirane, die mit Amino-, Sulfhydryl- oder phenolischen Hydroxyl-Gruppen reagieren können;
• vii) Chlor-enthaltende Derivate von s-Triazinen, der sehr reaktiv gegenüber Nukleophilen, wie Amino-, Sulfhydryl- und Hydroxy-Gruppen sind;
• viii) Aziridine, die auf s-Triazin-Verbindungen basieren, die oben im Detail angegeben sind, z.B. wie beschrieben von Ross, W. C. J. in Adv. Cancer Res. (1954) 2, 1, die mit Nukleophilen, wie Amino-Gruppen durch Ringöffnung reagieren.
• ix) Quadratsäure-Diethylester, wie beschrieben von Tietze, L. F. in Chem. Ber. (1991) 124, 1215; und
• x) α-Haloalkylether, die reaktivere Alkylierungsmittel sind als normale Alkylhalogenide, wegen der Aktivierung, die vom Ether-Sauerstoff-Atom verursacht wird, z.B. wie beschrieben von Benneche, T. et al. in Eur. J. Med. Chem. (1993) 28, 463.

Examples of reactive moieties capable of reacting with amino groups include alkylating and acylating agents. Representative alkylating agents include:

• i) α-haloacetyl compounds exhibiting specificity to amino groups in the absence of reactive thiol groups and of the X-CH ₂ CO- type (where X = Cl, Br or I), eg as described by Wong, YH. H. in Biochemistry (1979) 24, 5337;
• ii) N-maleimide derivatives which can react with amino groups, either via a Michael-type reaction or by acylation by addition to the ring carbonyl group as described by Smyth, DG et al. in J. Am. Chem. Soc. (1960) 82, 4600 and Biochem. J. (1964) 91, 589;
• iii) aryl halides, such as reactive nitrohalo-aromatic compounds;
• iv) alkyl halides as described by McKenzie, JA et al. in J. Protein Chem. (1988) 7, 581;
• v) aldehydes and ketones capable of Schiff-base formation with amino groups, the adducts formed usually being stabilized by reduction to give a stable amine;
• vi) epoxide derivatives, such as epichlorohydrin and bisoxiranes, which can react with amino, sulfhydryl or phenolic hydroxyl groups;
• vii) Chlorine-containing derivatives of s-triazines, which are highly reactive towards nucleophiles such as amino, sulf are hydryl and hydroxy groups;
• viii) aziridines based on s-triazine compounds detailed above, for example as described by Ross, WCJ in Adv. Cancer Res. (1954) 2, 1, which react with nucleophiles such as amino groups by ring opening react.
• ix) squaric acid diethyl ester as described by Tietze, LF in Chem. Ber. (1991) 124, 1215; and
• x) α-haloalkyl ethers which are more reactive alkylating agents than normal alkyl halides, because of the activation caused by the ether oxygen atom, eg as described by Benneche, T. et al. in Eur. J. Med. Chem. (1993) 28, 463.

• i) Isocyanate und Isothiocyanate, insbesondere aromatische Derivate, die stabile Harnstoff- bzw. Thioharnstoff-Derivate bilden und verwendet worden sind für das Vernetzen von Protein, wie beschrieben von Schick, A. F. in J. Biol. Chem. (1961) 236, 2477;
• ii) Sulfonylchloride, die von Herzig, D. J. et al. in Biopolymers (1964) 2, 349 beschrieben worden sind, und die nützlich sein können für das Einführen einer fluoreszierenden Reporter-Gruppe in den Linker;
• iii) Säurehalogenide
• iv) Aktive Ester, wie Nitrophenylester oder N-Hyroxysuccinimidylester;
• v) Säureanhydride, wie gemischte, symmetrische oder N-Carboxyanhydride;
• vi) andere nützliche Reagenzien für eine Amidbindungs-Bildung, wie beschrieben von Bodansky M. et al. in „Principles of Peptide Synthesis" (1984) Springer-Verlag;
• vii) Acylazide, z.B. wobei die Azid-Gruppe aus einem vorgeformten Hydrazid-Derivat unter Verwenden von Natriumnitrit erzeugt wird, z.B. wie beschrieben von Wetz, K. et al., ein Anal. Biochem. (1974) 58, 347;
• viii) Azlactone, die an Polymere gebunden sind, wie Bisacrylamid, z.B. wie beschrieben von Rasmussen, J. K. in Reactive Polymers (1991) 16, 199; und
• ix) Imidoester, die stabile Amidine bei Reaktion mit Amino-Gruppen bilden, z.B. wie beschrieben von Hunter, M. J. und Ludwig, M. L. in J. Am. Chem. Soc. (1962) 84, 3491.

Representative amino-reactive acylating agents include:

• i) isocyanates and isothiocyanates, especially aromatic derivatives which form stable urea or thiourea derivatives and have been used for cross-linking protein as described by Schick, AF in J. Biol. Chem. (1961) 236, 2477;
• ii) sulfonyl chlorides described by Herzig, DJ et al. in Biopolymers (1964) 2, 349, and which may be useful for introducing a fluorescent reporter group into the linker;
• iii) Acid halides
• iv) Active esters, such as nitrophenyl esters or N-hydroxysuccinimidyl esters;
• v) acid anhydrides, such as mixed, symmetrical or N-carboxyanhydrides;
• vi) other useful reagents for amide bond formation as described by Bodansky M. et al. in "Principles of Peptide Synthesis" (1984) Springer-Verlag;
• vii) acylazides, eg wherein the azide group is generated from a preformed hydrazide derivative using sodium nitrite, eg as described by Wetz, K. et al., Anal. Biochem. (1974) 58, 347;
• viii) azlactones bound to polymers such as bisacrylamide, eg as described by Rasmussen, JK in Reactive Polymers (1991) 16, 199; and
• ix) imidoesters which form stable amidines upon reaction with amino groups, for example as described by Hunter, MJ and Ludwig, ML in J. Am. Chem. Soc. (1962) 84, 3491.

Carbonyl groups, like aldehyde functions, can be reacted with weak protein bases at a pH such that nucleophilic protein side-chain functions are protonated. Weakness Bases include 1,2-aminothiols, such as those in N-terminal cysteine residues which are selectively stable 5-membered thiazolidine rings with aldehyde groups, e.g. as described by Ratner, S. et al. in J. Chem. Soc. (1937) 59, 200. Other weak bases, such as Phenylhydrazones can be used, e.g. as described by Heitzmann, H. et al. in proc. Natl. Acad. Sci. USA (1974) 71, 3537.

aldehydes and ketones can also be reacted with amines to form Schiff bases that be advantageously stabilized by reductive amination can.

Alkoxylamino units react easily with ketones and aldehydes to produce stable alkoxamines to produce, e.g. as described by Webb et al. in bioconjugates Chem. (1990) 1, 96.

Examples reactive units capable of reacting with carboxyl groups, include diazo compounds such as diazoacetate esters and diazoacetamides, those with high specificity react to produce ester groups, e.g. as described by Herriot, R.M. in Adv. Protein Chem. (1947) 3, 169. Carboxylic acid modifying Reagents, such as carbodiimides, formed by O-acyl urea formation, Reacted by amide bond formation can also be usefully employed; the Binding can be simplified by adding an amine or may lead to direct vector-receptor coupling. Useful water-soluble carbodiimides include 1-cyclohexy-3- (2-morpholinyl-4-ethyl) carbodiimide (CMC) and 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide (EDC), e.g. as described by Zot, H.G. and Puett, D. in J. Biol. Chem. (1989) 264, 15552. Other useful carboxylic acid modifiers Reagents include isoxazolium derivatives such as Woodwards reagent K; chloroformates, such as p-nitrophenyl chloroformate; Carbonyldiimidazoles, such as 1,1'-carbonyldiimidazole; and N-carbalkoxydihydroquinolines such as N- (ethoxycarbonyl) -2-ethoxy-1,2-dihydroquinoline.

Other potentially useful reactive moieties include vicinal diones, such as p-phenylenediglyoxal, which can be used to react with guanidyl groups, eg, as described by Wagner et al. in Nucleic Acid Res. (1978) 5, 4065; and diazonium salts capable of undergoing electrophilic substitution reactions, for example as described by Ishizaka, K. and Ishizaka T. in J. Immunol. (1960) 58, 163. Bis-diazonium compounds are readily prepared by treating aryl diamines with sodium nitrite in acidic solutions. It will be appreciated that functional groups in the reporter and / or vector, if desired, can be converted to other functional groups prior to the reaction, eg, to confer additional reactivity or selectivity. Examples of methods that are useful for this purpose include the conversion of amines to carboxylic acids using reagents such as dicarboxylic acid anhydrides; Converting amines to thiols using reagents such as N-acetylhomocysteine thiolactone, S-acetylmercaptosuccinic anhydride, 2-iminothiolane or thiol-containing succinimidyl derivatives; Conversion of thiols to carboxylic acids using reagents such as α-haloacetates; Conversion of thiols to amines using reagents such as ethyleneimine or 2-bromoethylamine; Conversion of carboxylic acids to amines using reagents such as carbodiimides followed by diamines; and converting alcohols to thiols using reagents such as tosyl chloride followed by transesterification with thioacetate and hydrolysis to the thiol with sodium acetate.

Vector Reporter couple can also be effected using enzymes such as zero-length crosslinking agent; thus, e.g. Transglutaminase, peroxidase and xanthine oxidase used, to create networked products. Reverse proteolysis can also used for crosslinking by amide bond formation.

Non-covalent Vector reporter coupling can e.g. by electrostatic charge interactions be effected, e.g. between a polylysinyl-functionalized Reporter and a polyglutamyl-functionalized vector, by Chelation in the form of stable metal complexes or by high affinity binding interaction, like avidin / biotin binding.

One Vector containing a peptide, lipo-oligosaccharide or lipopeptide linker or is coupled to it, which contains an element that capable of mediating membrane insertion may also be useful. An example is described by Leenhouts, J.M. et al. in Feb's Letters (1995) 370 (3), 189-192.

Couple may also be effected using avidin or streptavidin, the four high affinity binding sites for biotin have. Avidin can therefore be used to vector to reporters if both the vector and the reporter are biotinylated are. Examples are described by Bayer E.A. and Wilchek, M. in Methods Biochem. Anal. (1980) 26, 1. This method can also be expanded to include the reporter's tying to reporters Process that increases the association of the agent and the subsequent possible increased effectiveness promotes. Alternatively, avidin or streptavidin can be directly attached to the surface of the obliterator Be bound to reporter particles.

Noncovalent coupling may also utilize the bifunctional nature of bispecific immunoglobulins. These molecules can specifically bind and bind two antigens. For example, either bispecific IgG or chemically engineered bispecific F (ab) ' ₂ fragments can be used as the binding agent. Heterobifunctional bispecific antibodies have also been reported to bind two different antigens, eg as described by Bode C. et al. in J. Biol. Chem. (1989) 264, 944 and by Staerz, UD et al. in proc. Natl. Acad. Sci. USA (1986) 83, 1453. Similarly, any reporter and / or vector containing two or more antigenic determinants (eg as described by Chen, Aa et al., Am. J. Pathol. (1988) 130, 216 ) are crosslinked by antibody molecules and result in the formation of crosslinked arrays of Formula I agents of potentially enhanced efficacy.

So-called Zero-length binding agent a direct covalent bonding of two reactive chemical Induce groups without additional to introduce connecting material (For example, as in amide bond formation using carbodiimides or induced enzymatically) may, if desired, according to the invention used as well as agents, such as biotin / avidin systems, which induce non-covalent reporter-vector binding, and agents induce electrostatic interactions.

At the common however, the linking agent becomes two or more reactive units include, e.g. as described above, connected by a spacer element. The presence of such a spacer allows bifunctional Linkers, with specific functional groups within one molecule or to react between two different molecules, resulting in one Binding between these two components and inserting extrinsic linker-derived material into the reporter-vector conjugate leads.

The reactive units in a connecting agent may be the same (homobifunctional agents) or different (heterobifunctional Means or, where several dissimilar reactive units are present, heteromultifunctional agents) be a variety of possible Provide reagents that have a covalent bond between any cause chemical species can, either intramolecular or intermolecular.

The nature of extrinsic material introduced by the connective agent can be a critic have an influence on the targeting ability and overall stability of the final product. Thus, it may be desirable to introduce labile bonds, eg containing spacer arms, which are biodegradable or chemically sensitive or which incorporate enzymatic cleavage sites. Alternatively, the spacer may contain polymeric components, for example, to act as surfactants and enhance the stability of the agent. The spacer may also contain reactive moieties, eg as described above, to enhance surface crosslinking.

spacer elements can typically consist of aliphatic chains that effectively the reactive units of the linker by distances between 5 and 30 Å separate. You can also include macromolecular structures such as poly (ethylene glycols). Such polymeric structures, hereinafter referred to as PEGs are simple, neutral polyethers, which much attention in biotechnical and biomedical applications (See, e.g., Milton Harris, J. (Hsg) "Poly (ethylene glycol) chemistry, biotechnical and biomedical applications "Plenum Press, New York, 1992). PEGs are soluble in most solvents, including Water, and become in watery Environments strongly hydrated, with two or three water molecules attached to each Ethylene glycol segment is bound; this has the effect of Preventing adsorption of either other polymers or of Proteins on PEG-modified surfaces. PEGs are known that they are not toxic and active proteins or cells are not damage, while covalent bound PEGs for it are known to be non-immunogenic and non-antigenic. In addition, PEGs easily modified and bound to other molecules with little effect on their chemistry. Your advantageous solubility and biological properties are apparently of the many possible uses the PEGs and copolymers thereof, including block copolymers such as PEG polyurethanes and PEG polypropylenes.

suitable Molecular weights for PEG spacers according to the invention can be used, e.g. between 120 daltons and 20 kdaltons.

Of the Main mechanism for the uptake of particles by the cells of the reticuloendothelial system (RES) is opsonization by plasma proteins in the blood; mark them Foreign particles, which are then absorbed by the RES. The biological Properties of PEG Spacer Elements Used According to the Invention can to serve the circulation time of the remedy in a similar way Way to the one for PEGylated liposomes are observed to increase (see, e.g., Klibanov, A.L. et al. in FEBS Letters (1990) 268, 235-237 and Blume, G. and Cevc, G. in Biochim. Biophys. Acta (1990) 1029, 91-97). Increased coupling efficiency on surfaces of interest also be achieved using antibodies that conform to the terms of PEG spacers (see, e.g., Maruyama, K. et al., Biochim. Acta (1995) 1234, 74-80 and Hansen, C.B. et al. in Biochim. Biophys. Acta (1995) 1239, 133-144).

Other representative Spacer elements include polysaccharides of the structural type, like polygalacturonic acid, Glycosaminoglycans, heparinoids, cellulose and marine polysaccharides, such as alginates, chitosans and carrageenans; Memory-type polysaccharides, like strength, Glycogen, dextran and aminodextranes; Polyamino acids and methyl and ethyl esters of which, as in homo- and copolymers of lysine, glutamic acids and aspartic acid; and polypeptides, oligosaccharides and oligonucleotides, the enzyme cleavage sites can contain or can not contain.

In general, spacer elements may contain cleavable groups such as vicinal glycol, azo, sulfone, ester, thioester, or disulfide groups. Spacer, the biodegradable methylene diester or diamide groups of the formula - (Z) _m × Y × X C (R ¹ R ² ) × X × Y × (Z) _n - [wherein X and Z are selected from -O-, -S- and -NR- (wherein R is hydrogen or an organic group); each Y is a carbonyl, thiocarbonyl, sulfonyl, phosphoryl, or similar acid-forming group: m and n are each 0 or 1; and R ¹ and R ² each represent hydrogen, an organic group or a group -X.Y. (Z) _m -, or together form a divalent organic group], may also be useful; as discussed in eg WO-A-9217436, such groups are readily degradable in the presence of esterases, eg in vivo, but are stable in the absence of such enzymes. They may therefore be advantageously linked to therapeutic agents to allow the slow release thereof:
Poly [N- (2-hydroxyethyl) methacrylamides] are potentially useful spacer materials because of their low interaction with cells and tissues (see, eg, Volfovd, I., Rihová, B. and VR and Vetvicka, P. in J. Bioact., Comp (1992) 7, 175-190). Work with a similar polymer, mainly consisting of the closely related 2-hydroxypropyl derivative showed that it was endocytosed by the mononuclear phagocyte system only to a fairly minor extent (see Goddard, P., Williamson, I., Bron, J., Hutchkinson, LE, Nicholls, J. and Petrak, K. in J. Bioct Compat Polym. (1991) 6, 4-24.).

• i) Copolymere von Methylmethacrylat mit Methacrylsäure; diese können erosionsanfällig sein (siehe Lee, P. I. in Pharm. Res. (1993) 10, 980) und die Carboxylatsubstituenten können einen höheren Grad des Quellens verursachen als mit neutralen Polymeren;
• ii) Blockcopolymere von Polymethacrylaten mit bioabbaubaren Polyestern (siehe z.B. San Roman, J. und Guillen-Garcia, P. in Biomaterials (1991) 12, 236–241);
• iii) Cyanoacrylate, d.h. Polymere von Estern von 2-Cyanoacrylsäure – diese sind bioabbaubar und wurden in der Form von Nanoteilchen für selektive Arzneimittelfreisetzung verwendet (siehe Forestier, F., Gerrier, P., Chaumard, C., Quero, A. M., Couvreur, P. und Labarre, C. in J. Antimicrob. Chemoter. (1992) 30, 173–179);
• iv) Polyvinylalkohole, die wasserlöslich sind und im allgemeinen als biokompatibel angesehen werden (siehe z.B. Langer, R. in J. Control. Release (1991) 16, 53–60);
• v) Copolymere von Vinylmethylether mit Maleinsäureanhydrid, die als bioerosionsanfällig angeführt wurden, (siehe Finne, U., Hannus, M. und Urtii, A. in Int. J. Pharm. (1992) 78, 237–241);
• vi) Polyvinylpyrrolidone, z.B. mit einem Molekulargewicht von weniger als ungefähr 25.000, die schnell durch die Nieren gefiltert werden (siehe Hespe, W., Meier, A. M. und Blankwater, Y. M. in Arnzeim.-Forsch./Drug Res. (1977) 27, 1158–1162);
• vii) Polymere und Copolymere von kurzkettigen aliphatischen Hydroxysäuren, wie Glykol-, Milch-, Butter-, Valerian- und Capronsäuren (siehe z.B. Carli, F. in Chim. Ind. (Milan) (1993) 75, 494–9), einschließlich Copolymere, die aromatische Hydroxysäuren einbauen, um ihre Zersetzungsrate zu vergrößern (siehe Imasaki, K., Yoshida, M., Fukuzaki, N., Asano, M., Kumakura, M. Mashimo, T., Yamanaka, H. und Nagai. T. in Int. J. Pharm. (1992) 81, 31–38);
• viii) Polyester, die aus alternierenden Einheiten von Ethylenglykol und Terephthalsäure bestehen, z.B. Dacron^R, die nicht zersetzbar sind, aber hoch-biokompatibel;
• ix) Blockcopolymere, umfassend bioabbaubare Segmente aliphatischer Hydroxysäure-Polymere (siehe z.B. Younes, H., Nataf, P. R., Cohn D., Appelbaum, Y. J., Pizov, G. und Uretzky, G. in Biomater. Artif. Cells Artif. Organs (1988) 16, 705–719), z.B. in Zusammenhang mit Polyurethanen (siehe Kobayashi, H., Hyon, S. H. und Ikada, Y. in „Watercurable and biodegradable prepolymers"-J. Biomed. Mater. Res. (1991) 25, 1481–1494);
• x) Polyurethane, die dafür bekannt sind, dass sie in Implantaten gut toleriert werden, und die mit flexiblen, weichen Segmenten kombiniert werden können, z.B. umfassend Poly(tetramethylenglykol), Poly(propylenglykol) oder Poly(ethylenglykol) und aromatische, harte Segmente, z.B. 4,4'-Methylenbis(phenylenisocyanat) (siehe z.B. Ratner, B. D., Johnston, A. B. und Lenk, T. J. in J. Biomed. Mater. Res: Applied Biomaterials (1987) 21, 59–90; Sa Da Costa, V. et al. in J. Coll. Interface Sci (1981) 780, 445–452 und Affrossmann, S. et al. in Clinical Materials (1991) 8, 25–31);
• xi) Poly(1,4-dioxan-2-one), die angesichts ihrer hydrolysierbaren Esterbindungen als bioabbaubare Ester betrachtet werden können (siehe z.B. Song, C. X., Cui, X. M. und Schindler, A. in Med. Biol. Eng. Comput (1993) 31, S 147–150), und die Glycolideinheiten enthalten können, um ihre Absorbierbarkeit zu verbessern (siehe Bezwada, R. S., Shalaby, S. W. und Newman, H. D. J. in Agricultural and synthetic polymers: Biodegradablity and utilization (1990) (Hrsg. Glass, J. E. und Swift, G.), 167–174 – ACS symposium Series, #433, Washington D. C., U.S.A.- American Chemical Society);
• xii) Polyanhydride, wie Copolymere von Sebacinsäure (Oktandisäure) mit Bis(4-carboxy-phenoxy)propan, die in Kaninchenstudien (siehe Brem, H., Kader, A., Epstein, J. I., Tamargo, R. J., Domb, A., Langer, R. und Leong, K. W. in Sel. Cancer Ther. (1989) 5, 55–65) und Rattenstudien (siehe Tamargo, R. J., Epstein, J. I., Reinhard, C. S., Chasin, M. und Brem, H. in J. Biomed. Mater. Res. (1989) 23, 253–266) als nützlich für eine gesteuerte Freisetzung von Arzneimitteln im Hirn ohne evidente toxische Effekte gezeigt worden sind;
• xiii) Bioabbaubare Polymere, die Ortho-Estergruppen enthalten, welche für die kontrollierte Freisetzung in vivo eingesetzt worden sind (siehe Maa, Y. F. und Heller, J. in J. Control. Release (1990) 14, 21–28); und
• xiv) Polyphosphazene, die anorganische Polymere sind, die aus alternierenden Phosphor- und Stickstoffatomen bestehen (siehe Crommen, J. H., Vandorpe, J. und Schacht, E. H. in J. Control. Release (1993) 24, 167–180).

Other potentially useful polymeric spacer materials include:

• i) copolymers of methyl methacrylate with methacrylic acid; these may be susceptible to erosion (see Lee, PI in Pharm. Res. (1993) 10, 980) and the carboxylate substituents may cause a higher degree of swelling than with neutral polymers;
• ii) block copolymers of polymethacrylates with biodegradable polyesters (see, eg, San Roman, J. and Guillen-Garcia, P. in Biomaterials (1991) 12, 236-241);
• iii) cyanoacrylates, ie polymers of esters of 2-cyanoacrylic acid - these are biodegradable and have been used in the form of nanoparticles for selective drug release (see Forestier, F., Gerrier, P., Chaumard, C., Quero, AM, Couvreur, P. and Labarre, C. in J. Antimicrob. Chemoter. (1992) 30, 173-179);
• iv) polyvinyl alcohols which are water-soluble and are generally considered biocompatible (see eg Langer, R. in J. Control, Release (1991) 16, 53-60);
• v) copolymers of vinylmethyl ether with maleic anhydride cited as being susceptible to bioerrosion (see Finn, U., Hannus, M. and Urtii, A. in Int. J. Pharm. (1992) 78, 237-241);
• vi) polyvinylpyrrolidones, eg having a molecular weight of less than about 25,000, which are rapidly filtered by the kidneys (see Hespe, W., Meier, AM and Blankwater, YM in Arnzeim. -Forsch./Drug Res. (1977) 27, 1158-1162);
• vii) Polymers and copolymers of short chain aliphatic hydroxy acids such as glycolic, lactic, butyric, valeric and capric acids (see eg Carli, F. in Chim. Ind. (Milan) (1993) 75, 494-9), including Copolymers incorporating aromatic hydroxy acids to increase their rate of decomposition (see Imasaki, K., Yoshida, M., Fukuzaki, N., Asano, M., Kumakura, M. Mashimo, T., Yamanaka, H., and Nagai. T. Int. J. Pharm. (1992) 81, 31-38);
• viii) polyesters consisting of alternating units of ethylene glycol and terephthalic acid, eg Dacron ^R , which are not decomposable but highly biocompatible;
• ix) block copolymers comprising biodegradable segments of aliphatic hydroxy acid polymers (see eg Younes, H., Nataf, PR, Cohn D., Appelbaum, YJ, Pizov, G. and Uretzky, G. in Biomater Artif. Cells Artif. 1988) 16, 705-719), for example in connection with polyurethanes (see Kobayashi, H., Hyon, SH and Ikada, Y. in "Watercurable and Biodegradable prepolymers" J. Biomed. Mater. Res. (1991) 25, 1481-1494);
• x) polyurethanes which are known to be well tolerated in implants and which can be combined with flexible, soft segments, eg comprising poly (tetramethylene glycol), poly (propylene glycol) or poly (ethylene glycol) and aromatic hard segments, for example, 4,4'-methylenebis (phenylene isocyanate) (see, for example, Ratner, BD, Johnston, AB and Lenk, TJ in J. Biomed., Mater Res: Applied Biomaterials (1987) 21, 59-90, Sa Da Costa, V. in J. Coll., Interface Sci (1981) 780, 445-452 and Affrossmann, S. et al., Clinical Materials (1991) 8, 25-31);
• xi) poly (1,4-dioxan-2-ones) which, in view of their hydrolyzable ester bonds, can be regarded as biodegradable esters (see, for example, Song, CX, Cui, XM and Schindler, A. in Med. Biol. Eng. Comput. 1993) 31, pp. 147-150), and which may contain glycolide units to improve their absorbability (see Bezwada, RS, Shalaby, SW and Newman, HDJ in Agricultural and Synthetic Polymers: Biodegradablity and Utilization (1990) (Ed. Glass , JE and Swift, G.), 167-174 - ACS symposium Series, # 433, Washington DC, USA - American Chemical Society);
• xii) polyanhydrides such as copolymers of sebacic acid (octanedioic acid) with bis (4-carboxy-phenoxy) propane used in rabbit studies (see Brem, H., Kader, A., Epstein, JI, Tamargo, RJ, Domb, A., Langer, R. and Leong, KW in Sel. Cancer Ther. (1989) 5, 55-65) and rat studies (see Tamargo, RJ, Epstein, JI, Reinhard, CS, Chasin, M. and Brem, H. in J Biomed. Mater. Res. (1989) 23, 253-266) have been shown to be useful for controlled release of drugs in the brain without evident toxic effects;
• xiii) Biodegradable polymers containing ortho-ester groups which have been used for controlled release in vivo (see Maa, YF and Heller, J. in J. Control, Release (1990) 14, 21-28); and
• xiv) polyphosphazenes which are inorganic polymers consisting of alternating phosphorus and nitrogen atoms (see Crommen, JH, Vandorpe, J. and Schacht, EH in J. Control, Release (1993) 24, 167-180).

The The following tables list connecting means that can be used in Means according to the invention useful are.

footnotes:

• (1) = iodable
• (2) = fluorescent

In addition to the straight-chain and branched PEG-like linkers already contemplated (eg, polyethylene glycol and others containing from 2 to 100 repeating units of ethylene oxide), linkers in the VLR system may independently be a chemical bond or the residue of a linking group. As used herein, the term "residue of a linking group" refers to a moiety which remains from, results of, or is derived from the reaction of a vector reactive group with a reactive site on a vector The term "vector reactive group As used herein refers to any group that can react with functional groups typically found on vectors whose derivatization only minimally affects the ability of the vector to bind to its receptor. However, it is specifically contemplated that such vector-reactive groups may also react with functional groups typically found on relevant protein molecules. So be in one Aspect, the linkers useful in the practice of this invention are derived from those groups capable of reacting with any relevant molecule comprising a vector as described above containing a reactive group, whether such a relevant molecule is a protein, to form a connecting group.

Preferred linking groups are derived from vector reactive groups selected from, but not limited to:
a group that will react directly with carboxy, aldehyde, amine (NHR), alcohols, sulfhydryl groups, activated methylenes, and the like on the vector, eg, active halogen-containing groups including, for example, chloromethylphenyl groups and chloroacetyl [ClCH ₂ C (= O) -] groups, activated 2- (substituted with a leaving group) ethylsulfonyl and ethylcarbonyl groups such as 2-chloroethylsulfonyl and 2-chloroethylcarbonyl; vinylsulfonyl; vinylcarbonyl; epoxy; isocyanato; isothiocyanato; Aldehyde; aziridine; succinimidoxycarbonyl; activated acyl groups, such as carboxylic acid halides; mixed anhydrides and the like.

A group that can easily react with modified vector molecules containing a vector-reactive group, ie, vectors containing a reactive group modified to contain reactive groups, such as those mentioned in (1) above, for example, by oxidation of the vector to an aldehyde or a carboxylic acid, in which case the "linking group" can be derived from reactive groups selected from amino, alkylamino, arylamino, hydrazino, alkylhydrazino, arylhydrazino, carbazido, semicarbazido, thiocarbazido, thiosemicarbazido , Sulfhydryl, sulfhydrylalkyl, sulfhydrylaryl, hydroxy, carboxy, carboxyalkyl and carboxyaryl The alkyl moieties of the linking groups may contain from 1 to about 20 carbon atoms The aryl moieties of the linking groups may contain from about 6 to about 20 carbon atoms;
a group which can be bonded to the vector containing a reactive group or to the modified vector as recited in (1) and (2) above by using a crosslinking agent. The residues of certain useful crosslinking agents, such as homobifunctional and heterobifunctional gelatin hardeners, bisepoxides and bisisocyanates, can become part of a linking group during the crosslinking reaction. However, other useful crosslinking agents may facilitate crosslinking, eg as consumable catalysts, and are not present in the final conjugate. Examples of such crosslinking agents are carbodiimide and carbamoylonium crosslinking agents as disclosed in U.S. Patent No. 4,421,847 and the ethers of U.S. Patent No. 4,877,724. With these crosslinkers, one of the reactants, such as the vector, must have a carboxyl group and the other, such as a long chain spacer, must have a reactive amine, alcohol or sulfhydryl group. In the amide bond formation, the crosslinking agent first reacts selectively with the carboxyl group, is then cleaved out during the reaction of the so "activated" carboxyl group with an amine to form an amide bond between the two units and so covalently bond The advantage of this approach is that the crosslinking of similar molecules, eg, vector to vector, is avoided while the reaction of, for example, homobifunctional crosslinkers is non-selective and undesirable crosslinked molecules are obtained.

Sulfo-SMCC

Sulfosuccinimidyl-4-(N-maleimidomethyl)cyclohexan-1-carboxylat

Sulfo-SIAB

Sulfosuccinimidyl-(4-iodacetyl)aminobenzoat

Sulfo-SMPB

Sulfosuccinimidyl-(4-(p-maleimidophenyl)butyrat

2-IT

2-Iminothiolan

SATA

N-Succinimidyl-S-acetylthioacetat

Preferred useful linking groups are derived from various heterobifunctional crosslinking reagents, such as those listed in Pierce Chemical Company Immunotechnology Catalog - Protein Modification Section, (1995 and 1996). Useful non-limiting examples of such reagents include:

Sulfo-SMCC

Sulfosuccinimidyl 4- (N-maleimidomethyl) cyclohexane-1-carboxylate

Sulfo-SIAB

Sulfosuccinimidyl (4-iodoacetyl) aminobenzoate

Sulfo-SMPB

Sulfosuccinimidyl (4- (p-maleimidophenyl) butyrate

2-IT

2-iminothiolane

SATA

N-succinimidyl-S-acetylthioacetate

In addition to the above description, all or part of the linking groups may also consist of and be derived from complementary sequences of nucleotides and residues of nucleotides, both naturally occurring and modified, preferably non-self-associating oligonucleotide sequences. Particularly useful, non-limiting reagents for the incorporation of modified nucleotide units containing reactive functional groups, such as amine and sulfhydryl groups, into an oligonucleotide sequence are commercially available from, for example, Clontech Laboratories Inc. (Palo Alto California) and include Uni-Link Amino Modifier (Catalog # 5190), Biotin-ON Phosphoramidite (Catalog # 5191), N-MNT C6 Amino Modifier (Catalog # 5202), Amino Modifier II (Catalog # 5203), DMT-C6-3'Amin-ON (Catalog # 5222), C6 thiol modifier (Catalog # 5211) and the like. In one aspect, the linking groups of this invention are derived from the reaction of a reactive functional group, such as an amino or Sulfhydryl group as available in the above-mentioned Clontech reagents, one or more of which has been incorporated into an oligonucleotide sequence, with, for example, one or more of the previously described vector reactive groups, such as a heterobifunctional group on the vector.

By Binding two complementary Oligonucleotide sequences, one to the vector and the other to the Reporter, includes the resulting double-stranded, hybridized Oligonucleotide then the linking group between the vector and the reporter.

Other polymer systems that serve as linkers include:
Poly (L or D or DL amino acids) = proteins and peptides; naturally occurring or synthetic
Pseudo-poly (amino acids) = (amino acids linked by non-amide bonds)
Poly (L- or D- or DL-lactide) and the copolymers eg poly (L-lactide / DL-lactide) poly (glycolide)
L-lactide / glycolide copolymers
Poly-, caprolactone and its copolymers
polyanhydrides
Poly (orthoester)
polyphosphazenes
Long chain straight chain or branched lipids (& phospholipids) sugars and
carbohydrates
Oligonucleotides (see above)
as well as mixtures of the above.

unifying Means according to the invention are used, generally connecting the vector to the reporter or the reporter to the re porter with some degree of specificity bring, and can also be used to one or more therapeutically active Means to bind.

The The present invention accordingly provides a tool for the therapeutic Drug administration in combination with vector-mediated Alignment of the product to the desired location. By "therapeutic" or "medicinal" is meant a means the cheap one Effect on a specific disease in a living human or non-human animal possesses.

therapeutic Compounds according to the present invention Invention can be used inside a molecular aggregate or particulate Linkers encapsulated or bound or incorporated into the encapsulating walls of a vesicular Encapsulated Linker. So can the therapeutic compound to a part of the surface be bound, e.g. above covalent or ionic bonds, or may be physically in one encapsulating material mixture, especially if the drug has a material-like polarity or solubility has to keep it from leaking out of the product before intended is that in the body acts. The release of the drug can only by wetting contact following with blood after administration or as a consequence other internal or external influences, e.g. Resolution processes, catalyzed by enzymes, or use in which the linker reporter is a gas-containing vesicle.

The therapeutic substance may be attached to the encapsulating membrane surface of a vesicular Linkers using a suitable binding agent bound be, e.g. as described here. So you can, for example initially Phospholipid derivative to which the drug by a / biodegradable bond or linker is bound, and then this Incorporate the derivative into the material used to make the vesicle membrane is used as described above. Alternatively, the agent initially without the therapeutic agent is then prepared, which is then particulate (e.g. vesicular) Means coupled before use or applied to it can. So could e.g. an adjuvant to a suspension of microbubbles in aqueous Be given media and shaken to attach or attach the therapeutic to the microbubbles allow.

The Therapeutic may e.g. be a drug or a pro-drug that for use in treating congestive heart failure or another cardiovascular Therapy is known.

By Targeting of an agent according to the invention, containing a prodrug-activating enzyme in pathological areas, you can map the targeting of the enzyme, which makes it possible to visualize if the agent is targeted correctly and if the mean of non-target areas has disappeared. In this way can you get the optimal time for determine the injection of a prodrug into individual patients.

A Another alternative is a pro-drug, a pro-drug-activating Enzyme and a vector in the same particulate linker / reporter to install such a way that the pro-drug only after one external stimulus will be activated. Such a stimulus can e.g. its a blasting of vesicles by external ultrasound, Light stimulation of a chromophore reporter or magnetic Heating a superparamagnetic reporter after the desired targeting has been achieved.

So-called Pro-drugs can in agents according to the invention be used. So can Drugs are derivatized to their physicochemical properties to change and to adapt the means of the invention; such derivatized Medicines can are considered as pro-drugs and are usually inactive until the Cleavage of the derivatizing group the active form of the drug regenerated.

therapeutics can in a simple manner according to the invention to the heart and the vasculature in general and to the liver, Spleen and the kidneys and other regions, such as the lymphatic system, body cavities or the gastrointestinal system will be delivered.

For example, where the reporter is a chelated metal species (e.g., a paramagnetic Metal ion or a metal radionuclide), the linker may comprise one Chain linked to a metal-chelating group, one polymeric chain having a plurality of metal-chelating groups, the molecular backbone hang or in the molecular backbone are incorporated, a branched polymer with metal chelating Groups at branch termini (e.g., a dendrimeric polychelant), etc. What kind of The linker required is that he has the vector and reporter units simply tied together for a reasonable amount of time. With a reasonable amount of time is meant a time sufficient for the contrast agent to be desired Effects, e.g. during the contrast in vivo during a diagnostic imaging procedure.

So It can be under certain circumstances he wishes be that the linker is biodegraded after administration. By selecting a suitable biodegradable linker, it is possible to reconcile the biodistribution and bioelimination patterns for to modify the vector and / or reporter. Where the vector and / or reporter are biologically active or capable are, unwanted To exert effects if they are stored after the imaging procedure is over, it may be desired to schedule biodegradability of biodegradation, which is a suitable bioelimination or metabolic degradation of the vector and / or reporter units ensures. So a linker can contain a biodegradable function, the decomposition products with modified biodistribution patterns which results from the release of the reporter from the vector or from follow the fragmentation of a macromolecular structure. through an example of Linkers carrying chelated metal ion reporters, it is possible that the linker has incorporated a biodegradable function that degrades releases a precipitable chelate compound, which is the reporter contains. Accordingly, can, if desired, biodegradable functions are incorporated within the linker structure, preferably at sites containing (a) branching sites, (b) at or near binding sites for Vectors or reporters; or (c) such that biodegradation is physiological tolerable or rapidly excreted fragments.

Examples suitable biodegradable functions include ester, amide, double ester, Phosphorus, ether, thioether, guanidyl, acetal and ketal functions.

As discussed above, the linker group, if desired, in its molecular backbone Groups have incorporated the biodistribution of the contrast agent or which has a suitable spatial conformation for the contrast agent ensure, e.g. to allow water access to chelated paramagnetic metal ion reporters. For example, the linker backbone partially or substantially all in one or more Polyalkylene oxide chains exist.

Thus, the linker can be considered as a composite of optionally biodegradable vector binding (V _b) - and reporter binding (R _b) groups joined via linker backbone (R _b) groups, which linker backbone groups linker side chain (L _sc ) Groups, to modify biodistribution, etc., and can incorporate biodegradable functions themselves. The R _b and V _b linking groups may be pendant from the linker backbone or may be at linker backbone termini, eg with an R _b or V _b group on a L _b termini, where R _b - or V _b - Groups bind together two L _b termini or wherein one L _b termini bears two or more R _b or V _b groups. The L _b and L _sc groups will favorably be oligomeric or polymeric structures (eg, polyesters, polyamides, polyethers, polyamines, oligopeptides, polypeptides, oligo- and polysaccharides, oligonucleotides, etc.), preferably structures having at least partially a hydrophilic or lipophilic one Nature, eg hydrophilic, amphiphilic or lipophilic structures.

Of the Linker may be of low, medium or high molecular weight be, e.g. up to 2 MD. Become linker with generally higher molecular weight be preferred if using a variety of vectors or reporters should be loaded, or if necessary vector and reporter separate from each other, or if the linker itself serves to play a role in the modification of biodistribution. Generally, however, linkers will be from 100 to 100,000 D, especially 120 D to 20 kD in molecular weight.

The Conjugation of the linker to the vector and the linker to the reporter may be by any suitable chemical conjugation technique take place, e.g. covalent bonding (e.g., ester or amide formation), metal chelation and another metal-coordinative or ionic bond, again as described above.

Examples Suitable linker systems include the enhancer-polychelant structures of US-A-5364613 and WO 90/12050, polyamino acids (e.g., polylysine), functionalized PEG, polysaccharides, glycosaminoglycans, dendritic polymers as described in WO 93/06868 and Tomalia et al. in Angew. Chem. Int. Ed. Engl. 29: 138-175 (1990), PEG chelating Polymers as described in WO 94/08629, WO 94/09056 and WO 96/26754, Etc.

Where the reporter is a chelated metal ion, the linker group will generally incorporate the chelating moiety. Alternatively, the chelated metal can be carried in or on a particular reporter. In either case, conventional metal chelating groups, as are well known in the fields of radiopharmaceuticals and MRI contrast media, may be used, for example linear, cyclic and branched polyaminopolycarboxylic acids and phosphoroxyacid equivalents, and other sulfur and / or nitrogen ligands known in the art, eg, DTPA, DTPA-BMA, EDTA, DO3A, TMT (see, eg, US-A-5367080), BAT and analogs (see, eg, Ohmono et al., J. Med. Chem. 35: 157 252: 326-332 (1984)), the N ₂ S ₂ chelator ECD of Neurolite, MAG (see Jurisson et al., Chem. 1137-1156 (1993), HIDA, DOXA (1-oxa-4,7,10-triazacyclododecane triacetic acid), NOTA (1,4,7-triazacyclononane triacetic acid), TETA (1,4,8,11-tetraazacyclotetradecane tetraacetic acid), THT 4 '- (3-amino-4-methoxy-phenyl) -6,8''- bis (N', N'-dicarboxymethyl-N-methylhydrazino) -2,2 ': 6', 2 '' - terpyridine), etc. In this regard, the reader is referred to the patent literature of Sterli Winthrop, Nycomed (including Nycomed Imaging and Nycomed Salutar), Schering, Mallinckrodt, Bracco and Squibb, which refer to chelating agents for diagnostic metals, eg MR, X-ray and radiodiagnostic agents. See, for example, US-A-4647447, EP-A-71564, US-A-4687659, WO 89/00557, US-A-4885363 and EP-A-232751.

reporter

The Reporter units in the contrast agents of the invention may have a be any unit for detection either directly or indirectly or in an in vivo diagnostic imaging procedure, e.g. Units containing detectable radiation (e.g., by radioactive Decay, fluorescence excitation, spin resonance excitation, etc.) or be induced to emit them, units that are local electromagnetic fields (e.g., paramagnetic, superparamagnetic, ferrimagnetic or ferromagnetic species), units which Absorb or scatter radiant energy (e.g., chromophores, particles (including gas or liquid containing vesicle), heavy elements and compounds thereof, etc.), and units producing a detectable substance (e.g. Gas microbubble generators) Etc.

A very broad range of materials detectable by diagnostic imaging moieties are known in the art and the reporter will be selected according to the imaging modality to be used. For example, for ultrasound imaging, an echogenic material or material capable of generating an echogenic material may normally be selected; for X-ray imaging, the reporter will generally be or contain a heavy atom (eg, a relative atomic mass of 38 or above). For MR imaging, the reporter will either be a non-zero nuclear spin isotope (such as ¹⁹ F) or a material with unpaired electron spins and therefore paramagnetic, superparamagnetic, ferrimagnetic or ferromagnetic properties, for light imaging the reporter will be a light scatterer (eg for a magnetometric imaging, the reporter will have detectable magnetic properties, for imaging with electrical impedance the reporter will affect electrical impedance and for scintigraphy, SPECT, PET etc. the reporter will become a radionuclide se in.

Examples of suitable reporters are widely known from the literature of diagnostic imaging, eg, magnetic iron oxide particles, gas-containing vesicles, chelated paramagnetic metals (such as Gd, Dy, Mn, Fe, etc.). See, for example, US-A-4647447, WO 97/25073, US-A-4863715, US-A-4770183, WO 96/09840, WO 85/02772, WO 92/17212, WO 97/29783, EP-A-554213, US Pat. No. 5,228,446, WO 91/15243, WO 93/05818, WO 96/23524, WO 96/17628, US Pat. 5387080, WO 95/26205, GB 9624918.0 , Etc.

Particularly preferred as reporters are: chelated paramagnetic metal ions such as Gd, Dy, Fe and Mn, especially when chelated by macrocyclic chelating groups (eg tetraazacyclododecane chelating agents such as DOTA, DO3A, HP-DO3A and analogs thereof) or by linker Chelating groups such as DTPA, DTPA-BMA, EDTA, DPDP, etc .; ^{Metal radionuclides} such as ⁹⁰ Y, ^99m Tc, ¹¹¹ In, ⁴⁷ Sc, ⁶⁷ / Ga, ⁵¹ Cr, ^177m Sn, ⁶⁷ Cu, ¹⁶⁷ Tm, ⁹⁷ Ru, ¹⁸⁸ Re, ¹⁷⁷ Lu, ¹⁹⁹ Au, ²⁰³ Pb and ¹⁴¹ Ce; superparamagnetic iron oxide crystals; Chromophores and fluorophores with absorption and / or emission maxima in the range of 300 nm to 1400 nm, especially 600 nm to 1200 nm, in particular 650 nm to 1000 nm; Vesicles containing fluorinated gases (ie containing the materials in the gaseous phase at 37 ° C containing fluorine, eg SF ₆ or perfluorinated C _1-6 hydrocarbons or other gases and gas precursors listed in WO 97/29783) ; chelated heavy metal cluster ions (eg, W or Mo polyoxy anions or the sulfur or mixed oxygen / sulfur analogs); covalently bonded non-metallic atoms that are either of high atomic number (eg, iodine) or radioactive, eg ¹²³ I, ¹³¹ I, etc .; atoms; an iodinated compound containing vesicles; Etc.

Generally expressed the reporter can be (1) a chelatable metal or a polyatomic Metal-containing ion (i.e., TcO, etc.) wherein the metal is a metal is a high atomic number (e.g., an atomic number greater than 37), a paramagnetic species (e.g., a transition metal or lanthanide), or a radioactive isotope, (2) a covalently bonded non-metallic Species that is an unpaired electron site (e.g., an oxygen or carbon in a stable free radical), a non-metal of a high atomic number or a radioisotope, (3) a polyatomic cluster or crystal, the Contains atoms with high atomic number and a cooperative magnetic Behavior (e.g., superparamagnetism, ferrimagnetism, or Ferromagnetism) or radionuclides, (4) a gas or a gas precursor (i.e., a material or mixture of materials which is gaseous at 37 ° C), (5) a chromophore (by which term species are included, which fluoresce or phosphoresce), e.g. an inorganic one or organic structure, in particular a complexed metal ion or an organic group with an extensive delocalized Electron system, or (6) a structure or group of properties, that change the electrical impedance, e.g. due to an extended delocalized electron system.

Examples particularly preferred reporter groups are described in more detail below.

Chelated metal reporters: Metal radionuclides, paramagnetic metal ions, fluorescent Metal ions, heavy metal ions and cluster ions

Preferred metal radionuclides include ⁹⁰ Y, ^99m Tc, ¹¹¹ In, ⁴⁷ Sc, ⁶⁷ Ga, ⁵¹ Cr, ^177m Sn, ⁶¹ Cu, ¹⁶⁷ Tm, ⁹⁷ Ru, ¹⁸⁸ Re, ¹⁷⁷ Lu, ¹⁹⁹ Au, ²⁰³ Pb and ¹⁴¹ Ce.

preferred paramagnetic metal ions include ions of transition and lanthanide metals (e.g. Metals with atomic numbers of 6-9, 21-29, 42, 43, 44 or 57-71), in particular Ions of Cr, V, Mn, Fe, Co, Ni, Cu, La, Ce, Pr, Nd, Pm, Sm, Eu, Gd, Tb, Dy, Ho, Er, Tm, Yb and Lu, in particular from Mn, Cr, Fe, Gd and Dy, more especially Gd.

preferred fluorescent metal ions include lanthanides, especially La, Ce, Pr, Nd, Pm, Sm, Eu, Gd, Tb, Dy, Ho, Er, Tm, Yb and Lu. Eu is especially preferred.

preferred Heavy metal-containing reporters can be atoms of Mo, Bi, Si and W included and can in particular polyatomic cluster ions (e.g., Bi compounds and W and Mo oxides) as described in WO 91/14460, WO 92/17215, WO 96/40287 and WO 96/22914.

The metal ions are desirably chelated by chelating groups on the linker moiety or in or on a particle (eg, a vesicle or a porous or non-porous inorganic or organic solid), especially linear, macrocyclic, terpyridine, and N ₂ S ₂ chelating agents, such as DTPA, DTPA-BMA, EDTA, DO3A, TMT. Further examples of suitable chelant groups are disclosed in US-A-4647447, WO 89/00557, US-A-5367080, US-A-5364613, etc.

The Linker moiety or the particle may be one or more such Chelating groups, if desired metallated by more as a metal species (e.g., to provide reporters that detectable in different imaging modalities).

Especially where the metal is non-radioactive, it is preferred that a polychelant linker or a particulate one Reporter is used.

One Chelating agent or a chelating group to which reference is made here can take the rest of one or more of a wide variety of chelating agents comprising a metal ion or a polyatomic ion (e.g., TcO).

As well known, a chelating agent is a compound, contains the donor atoms, which combine by coordinative bonding with a metal atom can, to form a cyclic structure that is a 0Chelationkomplex or chelated. This class of compounds is described in the Kirk-Othmer Encyclopaedia of Chemical Technology, Volume 5, 339-368.

The balance of a suitable chelating agent may be selected from polyphosphates, such as sodium tripolyphosphate and hexametaphosphoric acid, aminocarboxylic acids, such as ethylenediaminetetraacetic acid, N- (2-hydroxy) ethylenediaminetriacetic acid, nitrilotriacetic acid, N, N-di (2-hydroxyethyl) glycine, ethylene bis (hydroxyphenylglycine ) and diethylenetriaminepentaacetic acid; 1,3-diketones such as acetylacetone, trifluoroacetylacetone and thenoyltrifluoroacetone; Hydroxycarboxylic acids such as tartaric acid, citric acid, gluconic acid and 5-sulfosalicylic acid; Polyamines such as ethylenediamine, diethylenetriamine, triethylenetetraamine and triaminotriethylamine; Amino alcohols such as triethanolamine and N- (2-hydroxyethyl) ethylenediamine; aromatic heterocyclic bases such as 2,2'-diimidazole, picolinamine, dipicolinamine and 1,10-phenanthroline, phenols such as salicylaldehyde, disulfopyrocatechol and chromotopsic acid; Aminophenols such as 8-hydroxyquinoline and oximesulfonic acid; Oximes such as dimethylglyoxime and salicylaldoxime; Peptides containing proximal chelating functionality, such as polycysteine, polyhistidine, polyaspartic acid, polyglutamic acid, or combinations of such amino acids; Schiff bases such as disalicylaldehyde-1,2-propylenediimine, tetrapyrroles such as tetraphenylporphine and phthalocyanine; Sulfur compounds, such as toluenedithiol, meso-2,3-dimercaptosuccinic acid, dimercaptopropanol, thioglycolic acid, potassium ethylxanthate, sodium diethyldithiocarbamate, dithizone, diethyldithiophosphoric acid and thiourea, synthetic macrocyclic compounds, such as dibenzo [18] crown-6, (CH ₃ ) ₆ - [14] - 4,11] -diene-N ₄ and (2.2.2-cryptate); Phosphonic acids, such as nitrilotrimethylenephosphonic acid, ethylenediaminetetra (methylenephosphonic acid) and hydroxyethylidenediphosphonic acid, or combinations of two or more of the above mentioned agents. The residue of a suitable chelating agent preferably comprises a polycarboxylic acid group and preferred examples include: ethylenediamine-N, N, N ', N'-tetraacetic acid (EDTA); N, N, N ', N ", N" -diethylenetriamine pentaacetic acid (DTPA); 1,4,7,10-tetraazacyclododecane-N, N ', N'',N''' - tetraacetic acid (DOTA); 1,4,7,10-tetraazacyclododecane-N, N ', N "-triacetic acid (DO3A); 1-oxa-4,7,10-triazacyclododecane-N, N ', N "-triacetic acid (OTTA); Trans (1,2) -cyclohexanodiethylenetriamine pentaacetic acid (CDTPA).

Other suitable residues of chelating agents include proteins which for the Chelation of metals, such as technetium and rhenium, modified as described in U.S. Patent No. 5,078,985, the disclosure of which is hereby incorporated by reference incorporated herein by reference.

suitable Remnants of chelating agents can also derived from N3S and N2S2-containing compounds, such as. those described in U.S. Patent Nos. 4,444,690; 4670545, 4673562, 4897255, 4965392, 4980147, 4988496, 5021556 and 5075099 are.

Other suitable residues of chelating agents are described in WO 92/08494.

preferred chelating groups are selected from the group consisting of 2-aminomethylpyridine, imino acetic acid, iminodiacetic acid, ethylenediaminetetraacetic (EDTA), diethylenetriamine pentaacetic acid (DTPA), 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA), carbonyliminodiacetic acid, Methyleniminoessigsäure, methyleneiminodiacetic, Ethylenthioethyleniminoessigsäure, Ethylenthioethyleniminodiessigsäure, TMT, a terpyridinyl group, a chelating agent that a terpyridyl group and a carboxymethylamino group, or a salt of any of the above acids. specially preferred chelating groups are DTPA, DTPA-BMA, DPDP, TMT, DOTA and HPDO3A.

Representative chelating groups are also described in US 5559214 A WO 95/26754, WO 94/08624, WO 94/09056, WO 94/29333, WO 94/08624, WO 94/08629 A1, WO 94/13327 A1 and WO 94/12216 A1.

Methods of metalating any of the present chelating agents are within the skill of the art stands in technology. Metals can be incorporated into a chelating moiety by any of three general methods: direct incorporation, template synthesis and / or transmetalation. Direct installation is preferred.

So is it desirable that the metal ion easily complexed by the chelating agent is, e.g. by merely exposing or mixing an aqueous solution the unit containing the chelating agent with a metal set in an aqueous Solution, preferably having a pH in the range of about 4 to about 11. The Salt may be any salt, but preferred is the salt a water-soluble Salt of the metal, such as a halo salt, and more preferred are such Salts so selected that they do not bind the metal ion with the chelating Intermediate means. The containing a chelating agent Unit is preferably in aqueous solution at a pH of between about 5 to about 9, more preferably between a pH of about 6 to about 8. The moiety containing the chelating agent can be mixed with buffer salts such as citrate, acetate, phosphate and borate to the optimum pH to create. Preferably, the buffer salts are selected such that do not interfere with the subsequent binding of the metal ion to the chelating one Intermediate means.

at In diagnostic imaging, the vector-linker-reporter (VLR) construct is preferred a relationship of metal radionuclide ions to a chelating agent, which in such diagnostic imaging applications. In preferred embodiments is the mole ratio of metal ion per chelating agent about 1: 1000 to about 1: 1.

In radiotherapeutic applications, the VLR preferably contains a ratio of metal radionuclide ion to chelating agent that is effective in such therapeutic applications. In preferred embodiments, the molar ratio of metal ion per chelating agent is about 1: 100 to about 1: 1. The radionuclide can be selected, for example, from radioisotopes of Sc, Fe, Pb, Ga, Y, Bi, Mn, Cu, Cr, Zn, Ge, Mo, Ru, Sn, Sr, Sm, Lu, Sb, W, Re, Po , Ta and Tl. Preferred radionuclides include ⁴⁴ Sc, ⁶⁴ Cu, ⁶⁷ Cu, ²¹² Pb, ⁶⁸ Ga, ⁹⁰ Y, ¹⁵³ Sm, ²¹² Bi, ¹⁸⁶ Re and ¹⁸⁸ Re. Of these, ⁹⁰ Y is particularly preferred. These radioisotopes may be atomic or preferably ionic.

The following isotopes or isotope pairs can be used for both imaging and therapy without having to change the radiolabelling methodology or chelator: 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 .

Where the linker moiety contains a single chelant, that chelant may be attached directly to the vector moiety, eg, via one of the metal coordinating groups of the chelant containing an ester, amide, thioester, or thioamide bond with an amine, thiol, or hydroxyl group on the Vector can make. Alternatively, the vector and the chelating agent can be attached directly via functionality attached to the chelator backbone, eg, a CH ₂ -phenyl NCS group attached to a ring carbon of DOTA as proposed by Meares et al. in JACS 110: 6266-6267 (1988), or indirectly via a homo- or heterobifunctional linker, eg, a bis-amide, a bis-epoxide, a diol, a diacid, a difunctionalized PEG, etc. In that case the bifunctional linker conveniently provides a chain of 1 to 200, preferably 3 to 30, atoms between vector and chelator residue.

wobei Ch eine Chelatbildner-Einheit und Li eine Linkerrückgrat-Komponente ist, d.h. der Linker besitzt bevorzugt anhängende Chelatbildner, Chelatbildner im Rückgrat oder terminate Chelatbildner oder eine Kombination davon. Die anhängenden und im Rückgrat vorliegenden polymeren Strukturen können verzweigt sein, sind aber bevorzugter linear und die Wiederholungseinheiten (LiCh) oder andere Wiederholungseinheiten im Polymer können im Rückgrat befindliche oder anhängende, die Biodistribution modifizierende Gruppen aufweisen, z.B. Polyalkylengruppen, wie in WO 94/08629, WO 94/09056 und WO 96/20754. Die terminalen chelatbildenden Stukturen Li(CH)_n, die dendritische Polymere sein können wie in WO 93/06868, können die Biodistribution modifizierende Gruppen besitzen, die an Termini gebunden sind, die nicht durch Chelatbildner besetzt sind, und können den Bioabbau verstärkende Stellen innerhalb der Linkerstruktur wie in WO 95/28966 aufweisen.Where the linker moiety contains a plurality of chelant groups, the linker is or preferably contains portions of the formulas

wherein Ch is a chelating moiety and Li is a linker backbone component, ie, the linker preferably has pendant chelants, chelators in the backbone or terminal chelants, or a combination thereof. The pendant and backbone polymeric structures may be branched but are more preferably linear and repeat units (LiCh) or other repeating units in the polymer may have backbone or pendant biodistribution modifying groups, eg, polyalkylene groups as described in WO 94/08629. WO 94/09056 and WO 96/20754. The terminal chelating structures Li (CH) _n , which may be dendritic polymers as in WO 93/06868, may be the biodistribution have modifying groups attached to termini that are not occupied by chelators, and may have biodegradation enhancing sites within the linker structure as in WO 95/28966.

The Chelator moieties within the polychelant linker can via backbone functionalization of the chelating agent or by using one or more of the metal coordinating Groups of the chelating agent or by an amide or Etherbindungsbildung between the acid chelating agent and an amine or hydroxyl bearing linker backbone be, e.g. as in polylysine polyDTPA, polylysine polyDOTA and in the so-called amplifier polychelants WO 90/12050. Such polychelant linkers can be attached one or more vector groups either directly (e.g., under use of amine, acid or hydroxyl groups in the polychelant linker) or via a bifunctional linker compound as discussed above for monochelator linker, be conjugated.

Where the chelated species is characterized by a particulate (or molecular aggregate) z. B. vesicular) linker the chelate, e.g. an unbound mono or Polychelat (such as Gd DTPA-BMA or Gd HP-DO3A) be within of the particle is included, or it may be a mono- or polychelate be attached to the particle by either covalent bonding or by interaction of an anchor group (e.g., a lipophilic group) on the mono- / polychelate with the membrane of a vesicle (see for example WO 96/11023) is conjugated.

Nonmetallic atomic reporters

Preferred non-metallic atomic reporters include radioisotopes such as ¹²³ I and ¹³¹ I as well as non-zero nuclear spin atoms such as ¹⁸ F and heavy atoms such as I.

such Reporter, preferably a plurality thereof, e.g. 2 to 200, can be covalent to a linker backbone be bound, either directly using conventional chemical synthesis techniques or via a supporting group, e.g. a triiodophenyl group.

In an embodiment This invention specifically relates to the use of radioisotopes of iodine taken into consideration. For example, if the vector or the linker consists of substituents chemically linked by iodine in a reaction of the formation of a covalent bond substituted can be such as. Substituents containing a hydroxyphenyl functionality, such substituents by methods well known in the art be labeled with a radioisotope of iodine. The iodine species can used in therapeutic and diagnostic imaging applications become. Meanwhile can at the same time a metal in a chelating agent on the same vector linker also in either therapeutic or diagnostic Imaging applications are used.

As with the metal chelants discussed above, such non-metallic atomic reporters can be attached to the linker or carried in or on a particulate linker, eg, in a vesicle (see WO 95/26205 and US Pat GB 9624918.0 ).

left of the type described above in connection with the metal reporters can for non-metallic atomic reporters with the non-metallic atomic reporters or Groups carrying such reporters can be used, which the Take the place of some or all of the chelating groups.

organic chromophore or fluorophore reporter

Preferred organic chromophore and fluorophore reporters include extended delocalized electron system groups, eg cyanines, merocyanines, phthalocyanines, naphthalocyanines, triphenylmethines, porphyrins, pyrilium dyes, thiapyrilium dyes, squarylium dyes, croconium dyes, azulenium dyes, indoanilines, benzophenoxazinium Dyes, benzothiaphenothiazinium dyes, anthraquinones, naphthoquinones, indathrene, phthaloylacridones, trisphenoquinones, azo dyes, intramolecular and intermolecular charge transfer dyes and dye complexes, tropones, tetrazines, bis (dithiolene) complexes, bis (benzene dithiolate) Complex, iodoaniline dyes, bis (S, O-dithiolene) complexes, etc. Examples of suitable organic or metalated organic chromophores can be found in "Topics in Applied Chemistry: Infrared Sucking Dyes" Ed M. Matsuoka, Plenum, NY 1990 , "Topics in Applied Chemistry: The Chemistry and Application of Dyes," Wari ng et al., Plenum, NY, 1990, "Handbook of Fluorescent Probes and Research Chemicals" Haugland, Molecular Probes Inc, 1996, DE-A-4445065, DE-A-4326466, JP-A-3/228046, Narayanan et al. J. Org. Chem. 60: 2391-2395 (1995), Lipowska et al. Hete rocyclic comm. 1: 427-430 (1995), Fabian et al. Chem. Rev. 92: 1197 (1992), WO 96/23525, Strekowska et al. J. Org. Chem. 57: 4578-4580 (1992), WO (Axis) and WO 96/17628. Specific examples of chromophores which may be used include xylene cyanol, fluorescein, dansyl, NBD, indocyanine green, DODCI, DTDCI, DOTCI and DDTCI.

Especially preferred are groups, the absorption maxima between 600 and 1000 nm to avoid interference with hemoglobin absorption (e.g., xylene cyanol).

und in Tabelle III auf Seite 28 von Matsuoka (supra), d.h.

Chalcogenopyrylomethin-Farbstoffe, z.B. Verbindungen 12 auf Seite 31 von Matsuoka (supra), d.h.

wobei Y = Te, Se, O oder NR;
Monochalcogenopyrylomethin-Farbstoffe, z.B. Verbindungen 13 auf Seite 31 von Matsuoka (supra), d.h.

wobei n = 1 oder 2;
Pyrilium-Farbstoffe, z.B. Verbindungen 14 (X = O) auf Seite 32 von Matsuoka (supra), d.h.

wobei X = O, S oder Se;
Thiapyrilium-Farbstoffe, z.B. Verbindungen 15 auf Seite 32 und Verbindung 1 auf Seite 167 von Matsuoka (supra), d.h.

wobei n = 1 oder 2;
Squarylium-Farbstoffe, z.B. Verbindung 10 und Tabelle IV auf Seite 30 von Matsuoka (supra), d.h.

wobei X = CH=CH, Y = Hund R = Et,
X = S, Y = H und R = Et, und
X = CMe₂, Y = H und R = Me,
und Verbindung 6, Seite 26 von Matsuoka (supra), d.h.

wobei X = CH=CH, Y = H und R = Et;
Croconium-Farbstoffe, z.B. Verbindung 9 und Tabelle IV auf Seite 30 von Matsuoka (supra), d.h.

wobei X = CH=CH, Y = H und R = Et,
X = S, Y = H und R = Et,
X = CMe₂, Y = H und R = Me,
und Verbindung 7, Seite 26 von Matsuoka (supra), d.h.

wobei X = CH=CH, Y = H und R = Et,
Azulenium-Farbstoffe, z.B. Verbindung 8 auf Seite 27 von Matsuoka (supra), d.h.

Merocyanin-Farbstoffe, z.B. Verbindung 16, R = Me, auf Seite 32 von Matsuoka (supra), d.h.

Indoanilin-Farbstoffe, wie Kupfer- und Nickel-Komplexe von Indoanilin-Farbstoffen, z.B. Verbindung 6 auf Seite 63 von Matsuoka (supra), d.h.

wobei R = Et, R' = Me, M = Cu,
R = Et, R' = Me, M = Ni,
R = Me, R' = H, M = Cu, oder
R = Me, R' = H, M = Ni,
Benzo[a]phenoxazinium-Farbstoffe und Benzo[a]phenothiazinium-Farbstoffe, z.B. wie gezeigt auf Seite 201 von Matsuoka (supra), d.h.

wobei X = O oder S;
1,4-Diaminoanthrachinon(N-alkyl)-3'-thioxo-2,3-dicarboximide, z.B. Verbindung 20, auf Seite 41 von Matsuoka (supra)

Indanthren-Pigmente, z.B.

siehe Verbindung 21 auf Seite 41 von Matsuoka (supra);
2-Arylamino-3,4-phthaloylacridon-Farbstoffe, z.B. Verbindung 22 auf Seite 41 von Matsuoka (supra)

Trisphenochinon-Farbstoffe, z.B. Verbindung 23 auf Seite 41 von Matsuoka (supra)

Azo-Farbstoffe, z.B. der Monoazo-Farbstoff, Verbindung 2 auf Seite 90 von Matsuoka (supra), d.h.

wobei X = CH=C(CN)₂, R₁ = R₂ = Et, R₃ = R₄ = H,
X = C(CN)=C(CN)₂, R₁ = R₂ = Et, R₃ = R₄ = H, oder
X =

und Y = C=O, R₁ = R₂ = Et, R₃ = R₄ = H,
oder Y = SO₂, R₁ = H, R₂ = CH(Me)nBu, R₃ = OMe und R₄ = NHAc;
Azo-Farbstoffe, z.B. der Polyazo-Farbstoff, Verbindung 5 auf Seite 91 von Matsuoka (supra), d.h.

intramolekulare Charge-Transfer-Donor-Akzeptor-Infrarot-Farbstoffe, z.B. die Verbindungen 6 und 7 auf Seite 91 von Matsuoka (supra), d.h.

nicht-benzoide aromatische Farbstoffe, z.B. Verbindung 8, ein Tropon, auf Seite 92 von Matsuoka (supra), d.h.

Tetrazin-Radikal-Farbstoffe, z.B. Verbindung 9 auf Seite 92 von Matsuoka (supra), d.h.

in der X = p-Phenylen oder
X = p-Terphenylen, wie auch Verbindung 10 auf Seite 92 von Matsuoka (supra), d.h.

in der X = p-Biphenyl,
Kationische Salze von Tetrazin-Radikal-Farbstoffen, z.B. Verbindung 11 auf Seite 92 von Matsuoka (supra)

in der X = p-Phenylenel;
Intermolekulare Donor-Akzeptor-Charge-Transfer-Farbstoffe, z.B. CT-Komplexe der Verbindungen 13b und 14a bis 14c auf Seite 93 von Matsuoka (supra), d.h.

wobei X = CH=N-N(Ph)₂ im Donor und

• a) Y = CN, Z = NO₂
• b) Y = CN, Z = H oder
• a) Y = Cl, Z = NO₂ in dem Akzeptor;

Anthrachinon-Farbstoffe, z.B. Verbindungen 12 (X = S oder Se) auf Seite 38 von Matsuoka (supra), d.h.

wobei X = S oder Se und Y = Tetrachloro, Tetrabromo, 2,3-Dicarbonsäure, 2,3-Dicarbonsäureanhydrid oder 2,3-Dicarbonsäure-N-phenylimid;
Naphthochinon-Farbstoffe, z.B. Verbindungen 2, 3 und 4 auf Seite 37 von Matsuoka (supra), d.h.

metallierte Azo-Farbstoffe, wie Azo-Farbstoffe, die Nickel, Kobalt, Kupfer, Eisen und Mangan enthalten;
Phthalocyanin-Farbstoffe, z.B. Verbindung 1 in Tabelle II auf Seite 51 von Matsuoka (supra), z.B.

Naphthalocyanin-Farbstoffe, z.B. Verbindung 3 in Tabelle II auf Seite 51 von Matsuoka (supra), z.B.

Metall-Phthalocyanine, wie Phthalocyanine, die Aluminium, Silizium, Nickel, Zink, Blei, Cadmium, Mangan, Vanadium, Kobalt, Kupfer und Eisen enthalten, z.B. Verbindung 1 in Tabelle III auf Seite 52 von Matsuoka (supra), z.B.

in der z.B. M = Mg;
Metall-Naphthalocyanine, wie Naphthalocyanine, die Aluminium, Zink, Kobalt, Magnesium, Cadmium, Silizium, Nickel, Vanadium, Blei, Kupfer und Eisen enthalten, siehe Verbindung 3 in Tabelle III auf Seite 52 von Matsuoka (supra), z.B.

in der z.B. M = Mg;
Bis(dithiolen)-Metall-Komplexe, die ein Metallion umfassen, wie Nickel, Kobalt, Kupfer und Eisen, die an vier Schwefelatome in einem Bis(S,S'-bidentat)-Liganden-Komplex koordiniert sind, z.B. siehe Tabelle I auf Seite 59 von Matsuoka (supra)

wobei
R₁ = R₂ = CF₃, M = Ni,
R₁ = R₂ = Phenyl, M = Pd,
R₁ = R₂ = Phenyl, M = Pt,
R₁ = C4 bis C10-Alkyl, R₂ = H, M = Ni,
R₁ = C4 bis C10-Alkyl, R₂ = H, M = Pd,
R₁ = C4 bis C10-Alkyl, R₂ = H, M = Pt,
R₁ = R₂ = Phenyl, M = Ni,
R₁ = R₂ = p-CH₃-Phenyl, M = Ni,
R₁ = R₂ = p-CH₃O-Phenyl, M = Ni,
R₁ = R₂ = p-Cl-Phenyl, M = Ni,
R₁ = R₂ = p-CF₃-Phenyl, M = Ni,
R₁ = R₂ = 3,4-diCl-Phenyl, M = Ni,
R₁ = R₂ = o-Cl-Phenyl, M = Ni,
R₁ = R₂ = o-Br-Phenyl, M = Ni,
R₁ = R₂ = 3,4-diCl-Phenyl, M = Ni,
R₁ = R₂ = p-CH₃, M = Ni,
R₁ = R₂ = 2-Thienyl, M = Ni,
R₁ = p-(CH₃)₂N-Phenyl, R₂ = Phenyl, M = Ni, und
R₁ = p-(CH₃)₂N-Phenyl, R₂ = p-N₂N-Phenyl, M = Ni,
Bis(benzoldithiolat)-Metall-Komplexe, die ein Metallion umfassen, wie Nickel, Kobalt, Kupfer und Eisen, die an vier Schwefelatome in einem Ligandenkomplex koordiniert sind, z.B. siehe Tabelle III auf Seite 62 von Matsuoka (supra), d.h.

wobei
X = Tetramethyl, M = Ni,
X = 4,5-Dimethyl, M = Ni,
X = 4-Methyl, M = Ni,
X = Tetrachloro, M = Ni,
X = H, M = Ni,
X = 4-Methyl, M = Co,
X = 4-Methyl, M = Cu, und
X = 4-Methyl, M = Fe;
N,O-Bidentat-Indoanilin-Farbstoffe, die ein Metallion umfassen, wie Nickel, Kobalt, Kupfer und Eisen, die an zwei Stickstoff- und zwei Sauerstoffatome von zwei N,O-Bidentat-Indoanilin-Liganden koordiniert sind, z.B. Verbindung 6 in Tabelle IV auf Seite 63 von Matsuoka (supra), z.B.

wobei R = Et, R' = Me, M = Cu,
R = Et, R' = Me, M = Ni,
R = Me, R' = H, M = Cu, und
R = Me, R' = H, M = Ni,
Bis(S,O-dithiolen)-Metall-Komplexe, die ein Metallion umfassen, wie Nickel, Kobalt, Kupfer und Eisen, die an zwei Schwefelatome und zwei Sauerstoffatome in einem Bis(S,O-bidentat)-Liganden-Komplex koordiniert sind, z.B. siehe US-Patent 3,806,462, z.B.

a-Diimin-dithiolen-Komplexe, die ein Metallion umfassen, wie Nickel, Kobalt, Kupfer und Eisen, die an zwei Schwefelatome und zwei Imino-Stickstoffatome in einem gemischten S,S- und N,N-Bidentat-Diliganden-Komplex koordiniert sind, z.B. siehe Tabelle II auf Seite 180, zweitletzter von unten, von Matsuoka (supra) (siehe auch Japanische Patente: 62/39,682, 63/126,889 und 63/139,303), z.B.

und
Tris(a-diimin)-Komplexe, die ein Metallion umfassen, das an sechs Stickstoffatome in einem Triliganden-Komplex koordiniert ist, z.B. siehe Tabelle II auf Seite 180 von Matsuoka (supra), letzte Verbindung, (siehe auch Japanische Patente 61/20,002 und 61/73,902), z.B.Other such examples include:
Cyanine dyes: such as heptamethine cyanine dyes, eg compounds 4a to 4g. Table II on page 26 of Matsuoka (supra)

and in Table III on page 28 of Matsuoka (supra), ie

Chalcogenopyrylomethine dyes, eg compounds 12 on page 31 of Matsuoka (supra), ie

where Y = Te, Se, O or NR;
Monochalcogenopyrylomethine dyes, eg compounds 13 on page 31 of Matsuoka (supra), ie

where n = 1 or 2;
Pyrilium dyes, eg compounds 14 (X = O) on page 32 of Matsuoka (supra), ie

where X = O, S or Se;
Thiapyrilium dyes, eg compounds 15 on page 32 and compound 1 on page 167 of Matsuoka (supra), ie

where n = 1 or 2;
Squarylium dyes, eg Compound 10 and Table IV on page 30 of Matsuoka (supra), ie

where X = CH = CH, Y = dog R = Et,
X = S, Y = H and R = Et, and
X = CMe ₂ , Y = H and R = Me,
and Compound 6, page 26 of Matsuoka (supra), ie

where X = CH = CH, Y = H and R = Et;
Croconium dyes, eg Compound 9 and Table IV on page 30 of Matsuoka (supra), ie

where X = CH = CH, Y = H and R = Et,
X = S, Y = H and R = Et,
X = CMe ₂ , Y = H and R = Me,
and Compound 7, page 26 of Matsuoka (supra), ie

where X = CH = CH, Y = H and R = Et,
Azulenium dyes, eg Compound 8 on page 27 of Matsuoka (supra), ie

Merocyanine dyes, eg Compound 16, R = Me, on page 32 of Matsuoka (supra), ie

Indoaniline dyes, such as copper and nickel complexes of indoaniline dyes, eg Compound 6 on page 63 of Matsuoka (supra), ie

where R = Et, R '= Me, M = Cu,
R = Et, R '= Me, M = Ni,
R = Me, R '= H, M = Cu, or
R = Me, R '= H, M = Ni,
Benzo [a] phenoxazinium dyes and benzo [a] phenothiazinium dyes, eg as shown on page 201 of Matsuoka (supra), ie

where X = O or S;
1,4-diaminoanthraquinone (N-alkyl) -3'-thioxo-2,3-dicarboximides, eg Compound 20, on page 41 of Matsuoka (supra)

Indanthrene pigments, eg

see compound 21 on page 41 of Matsuoka (supra);
2-arylamino-3,4-phthaloylacridone dyes, eg Compound 22 on page 41 of Matsuoka (supra)

Trisphenoquinone dyes, eg Compound 23 on page 41 of Matsuoka (supra)

Azo dyes, eg the monoazo dye, Compound 2 on page 90 of Matsuoka (supra), ie

where X = CH = C (CN) ₂ , R ₁ = R ₂ = Et, R ₃ = R ₄ = H,
X = C (CN) = C (CN) ₂ , R ₁ = R ₂ = Et, R ₃ = R ₄ = H, or
X =

and Y = C = O, R ₁ = R ₂ = Et, R ₃ = R ₄ = H,
or Y = SO ₂ , R ₁ = H, R ₂ = CH (Me) n Bu, R ₃ = OMe and R ₄ = NHAc;
Azo dyes, such as the polyazo dye, Compound 5 on page 91 of Matsuoka (supra), ie

intramolecular charge transfer donor-acceptor infrared dyes, eg, the compounds 6 and 7 on page 91 of Matsuoka (supra), ie

non-benzoic aromatic dyes, eg Compound 8, a tropon, at page 92 of Matsuoka (supra), ie

Tetrazine radical dyes, eg Compound 9 on page 92 of Matsuoka (supra), ie

in the X = p-phenylene or
X = p-terphenylene, as well as compound 10 on page 92 of Matsuoka (supra), ie

in which X = p-biphenyl,
Cationic salts of tetrazine radical dyes, eg Compound 11 on page 92 of Matsuoka (supra)

in the X = p-phenylenel;
Intermolecular donor-acceptor charge transfer dyes, eg CT complexes of compounds 13b and 14a to 14c on page 93 of Matsuoka (supra), ie

where X = CH = NN (Ph) ₂ in the donor and

• a) Y = CN, Z = NO ₂
• b) Y = CN, Z = H or
• a) Y = Cl, Z = NO ₂ in the acceptor;

Anthraquinone dyes, eg compounds 12 (X = S or Se) on page 38 of Matsuoka (supra), ie

wherein X = S or Se and Y = tetrachloro, tetrabromo, 2,3-dicarboxylic acid, 2,3-dicarboxylic anhydride or 2,3-dicarboxylic acid-N-phenylimide;
Naphthoquinone dyes, eg compounds 2, 3 and 4 on page 37 of Matsuoka (supra), ie

metallized azo dyes such as azo dyes containing nickel, cobalt, copper, iron and manganese;
Phthalocyanine dyes, eg Compound 1 in Table II on page 51 of Matsuoka (supra), eg

Naphthalocyanine dyes, eg Compound 3 in Table II on page 51 of Matsuoka (supra), eg

Metal phthalocyanines such as phthalocyanines containing aluminum, silicon, nickel, zinc, lead, cadmium, manganese, vanadium, cobalt, copper and iron, eg Compound 1 in Table III on page 52 of Matsuoka (supra), eg

in the example M = Mg;
Metal naphthalocyanines such as naphthalocyanines containing aluminum, zinc, cobalt, magnesium, cadmium, silicon, nickel, vanadium, lead, copper and iron, see compound 3 in Table III on page 52 of Matsuoka (supra), eg

in the example M = Mg;
Bis (dithiolene) metal complexes comprising a metal ion such as nickel, cobalt, copper and iron coordinated to four sulfur atoms in a bis (S, S'-bidentate) ligand complex, eg see Table I below Page 59 by Matsuoka (supra)

in which
R ₁ = R ₂ = CF ₃ , M = Ni,
R ₁ = R ₂ = phenyl, M = Pd,
R ₁ = R ₂ = phenyl, M = Pt,
R ₁ = C ₄ to C 10 -alkyl, R ₂ = H, M = Ni,
R ₁ = C ₄ to C 10 -alkyl, R ₂ = H, M = Pd,
R ₁ = C ₄ to C 10 -alkyl, R ₂ = H, M = Pt,
R ₁ = R ₂ = phenyl, M = Ni,
R ₁ = R ₂ = p-CH ₃ -phenyl, M = Ni,
R ₁ = R ₂ = p-CH ₃ O-phenyl, M = Ni,
R ₁ = R ₂ = p-Cl-phenyl, M = Ni,
R ₁ = R ₂ = p-CF ₃ -phenyl, M = Ni,
R ₁ = R ₂ = 3,4-diCl-phenyl, M = Ni,
R ₁ = R ₂ = o-Cl-phenyl, M = Ni,
R ₁ = R ₂ = o-Br-phenyl, M = Ni,
R ₁ = R ₂ = 3,4-diCl-phenyl, M = Ni,
R ₁ = R ₂ = p-CH ₃ , M = Ni,
R ₁ = R ₂ = 2-thienyl, M = Ni,
R ₁ = p- (CH ₃ ) ₂ N-phenyl, R ₂ = phenyl, M = Ni, and
R ₁ = p- (CH ₃ ) ₂ N-phenyl, R ₂ = pN ₂ N-phenyl, M = Ni,
Bis (benzenedithiolate) metal complexes comprising a metal ion, such as nickel, cobalt, copper and iron, coordinated to four sulfur atoms in a ligand complex, eg see Table III on page 62 of Matsuoka (supra), ie

in which
X = tetramethyl, M = Ni,
X = 4,5-dimethyl, M = Ni,
X = 4-methyl, M = Ni,
X = tetrachloro, M = Ni,
X = H, M = Ni,
X = 4-methyl, M = Co,
X = 4-methyl, M = Cu, and
X = 4-methyl, M = Fe;
N, O-bidentate indoaniline dyes comprising a metal ion, such as nickel, cobalt, copper and iron, coordinated to two nitrogen and two oxygen atoms of two N, O-bidentate-indoaniline ligands, eg, compound 6 in Table IV on page 63 of Matsuoka (supra), eg

where R = Et, R '= Me, M = Cu,
R = Et, R '= Me, M = Ni,
R = Me, R '= H, M = Cu, and
R = Me, R '= H, M = Ni,
Bis (S, O-dithiolene) metal complexes comprising a metal ion, such as nickel, cobalt, copper and iron, coordinated to two sulfur atoms and two oxygen atoms in a bis (S, O-bidentate) ligand complex for example see US Patent 3,806,462, eg

α-Diimine dithiolene complexes comprising a metal ion, such as nickel, cobalt, copper and iron, coordinated to two sulfur atoms and two imino nitrogen atoms in a mixed S, S and N, N-bidentate diligand complex for example see Table II on page 180, second from the bottom, from Matsuoka (supra) (see also Japanese patents: 62 / 39,682, 63 / 126,889 and 63 / 139,303), eg

and
Tris (α-diimine) complexes comprising a metal ion coordinated to six nitrogen atoms in a triligand complex, eg, see Table II on page 180 of Matsuoka (supra), final compound, (see also Japanese Patent 61 / 20,002 and 61 / 73,902), eg

Representative examples of visible dyes include fluorescein derivatives, rhodamine derivatives, coumarins, azo dyes, metalatable dyes, anthraquinone dyes, benzodifuranone dyes, polycyclic aromatic carbonyl dyes, indigoid dyes, polymethine dyes, azacarbocyanine dyes, hemicyanine dyes, barbituates, diazahemicyanine dyes , Styryl dyes, diaryl carbonium dyes, triaryl carbonium dyes, phthalocyanine dyes, quinophthalone dyes, triphenodioxazine dyes, formazan dyes, phenothiazine dyes such as methylene blue, azure A, azure B and azure C, oxazine dyes, thiazine Dyes, naphtholactam dyes, diazahemicyanine dyes, azopyridone dyes, azobenzene dyes, mordant dyes, acid dyes, basic dyes, metallated and pre-metalated dyes, xanthene dyes, direct dyes, leuco dyes that can be oxidized to dyes to produce with shades that bathochromes are shifted from those of the precursor leuco dyes, and other dyes such as those listed by Waring, DR and Hallas, G. in "The Chemistry and Application of Dyes, Topics in Applied Chemistry, Plenum Press, New York, NY, 1990. Additional dyes can be found listed in Haugland, RP, Handbook of Fluorescent Probes and Research Chemicals, 6th Edition, Molecular Probes , Inc., Eugenen OR, 1996.

such Chromophores and fluorophores can covalently either directly to the vector or within a linker structure be bound. Once again Linker of the type described above along with the metal reporters for organic Chromophores or fluorophores are used with the chromophores / fluorophores Be the place of some or all chelating groups taking.

As with the metal chelators discussed above, chromophores / fluorophores in or on particulate Be carried by linker units, e.g. in or on a vesicle, or covalently bonded to inert matrix particles, also called a light scattering reporter can act.

Particulate reporter or linker reporters

The particulate reporters and linker reporters generally fall into two categories-those in which the particle comprises a matrix or shell carrying or containing the reporter, and those in which the particle matrix itself is the reporter. Examples of the first category are: vesicles (eg, micelles, liposomes, microballoons, and microbubbles) containing a liquid, gas, or solid phase containing the contrast-effective reporter, eg, an echogenic gas or a precursor therefor (see, eg, US Pat GB 9700699.3 ), a chelated paramagnetic metal or radionuclide or a water-soluble iodinated X-ray contrast agent; porous particles loaded with the reporter, eg, molecular sieve particles loaded with a paramagnetic metal; and solid particles, eg, an inert biotolerant polymer, on which the reporter is bound or layered, eg, dye-loaded polymer particles.

Examples the second category are: light-scattering organic or inorganic Particles, magnetic particles (i.e., superparamagnetic, ferromagnetic or ferrimagnetic particles); and dye particles.

Preferred particulate reporters or reporter linkers include superparamagnetic particles (see US-A-4770183, WO 97/25073, WO 96/09840, etc.), echogenic vesicles (see WO 92/17212, WO 97/29783, etc.), Iodine-containing vesicles (see WO 95/26205 and GB 9624918.0 ) and dye-loaded polymer particles (see WO 96/23524).

The particulate reporters may have one or more vectors that are directly or indirectly bound to their surfaces. In general, it will be preferable to bind a plurality (eg, 2 to 50) of vector units per particle. In addition to the desired targeting vector, it will be particularly advantageous to also bind the flow-slowing vectors to the particles, ie vectors which have an affinity for the capillary lumen or other organ surfaces sufficient to allow passage of the contrast agent through the capillaries of the target organ, but is not sufficient in itself to immobilize the contrast agent. Such flow-slowing vectors (described, for example, in US Pat GB 9700699.3 ) may also serve to anchor the contrast agent once bound to its target site.

The Means by which vector-to-particle binding is achieved will depend on the nature of the particle surface. For inorganic particles can binding to the particle e.g. by means of an interaction between a metal-binding group (e.g., a phosphate, phosphonate, or oligo- or polyphosphate group) on the vector or on one Linker, which is bound to the vector done. For organic (e.g., polymeric) particles, vector attachment may be by means of a direct covalent bond between groups on the particle surface and reactive groups in the vector, e.g. Amide or ester bond, or by covalent attachment of the vector and the particle to a linker. Linkers of the type discussed above associated with chelated Metal reporters can although generally the linkers are not for coupling together be used by particles.

For non-particulate solids, eg, drops (eg of water-insoluble iodinated liquids as described in US-A-5318767, US-A-5451393, US-A-5352459 and US-A-5569448) and vesicles, the linker may desirably be hydrophobic For example, "anchor" groups, such as saturated or unsaturated C _12-30 chains, permeate the particle surface and bind the vector to the particle, so for phospholipid vesicles, the linker can be used to covalently attach the vector to a phospholipid that reacts with Examples of linker binding to vesicles and inorganic particles are described in U.S. Pat GB 9622368.0 and WO 97/25073.

Except the Vectors can other groups attached to the particle surface, e.g. stabilizers (to prevent aggregation) and biodistribution modifiers like PEG. Such groups are e.g. discussed in WO 97/25073, WO 96/09840, EP-A-284549 and US-A-4904479.

Preferably For example, the V-L-R agents of the invention will be nonpeptidic vectors have (as losartan), directly or indirectly to a reporter coupled, e.g. with covalently bound iodine radioisotopes, with metal chelates directly or via an organic linker group bound or attached to a particulate Reporter or linker reporter coupled, e.g. superparamagnetic Crystals (optionally coated, e.g., as in WO 97/25073) or a vesicle, e.g. a gas-containing or / an iodinated contrast agent containing micelle, liposome or microballoon.

The Means of the invention can administered to patients for imaging in amounts sufficient are to the desired Contrast with the special imaging technique. Where the Reporter is a metal, are generally dosages of 0.001 to 5.0 mmol of a chelated imaging metal ion per kilogram of a patient's body weight effective to provide adequate contrast enhancements to reach. For Most MRI applications will use the preferred dosages of Imaging metal ion in the range of 0.02 to 1.2 mmol / kg body weight lie while for X-ray applications Dosages of 0.05 to 2.0 mmol / kg are generally effective, about X-ray attenuation too to reach. Preferred dosages for most X-ray applications are from 0.1 to 1.2 mmol of the lanthanide or heavy metal compound per kg Body weight. Where the reporter is a radionuclide, dosages of 0.01 to 100 mCi, preferably 0.1 to 50 mCi normally per 70 kg of body weight be enough. Where the reporter is a superparamagnetic particle is, the dosage is usually 0.5 to 30 mg Fe / kg body weight be. Where the reporter is a gas or gas generator, e.g. in a microballoon, the dosage is usually 0.05 to 100 μl of gas per 70 kg body weight be.

The Dosage of the compounds of the invention for therapeutic use will depend on the condition being treated but in general will they are of the order of magnitude from 1 pmol / kg to 1 mmol / kg body weight lie.

The Compounds of the present invention can be used with conventional pharmaceutical or veterinary Auxiliaries may be formulated, e.g. Emulsifiers, fatty acid esters, Gelling agents, stabilizers, antioxidants that adjust osmolality Agents, buffers, pH adjusting agents, etc. and can be used in in a form suitable for parenteral or enteral administration are suitable, e.g. injection or infusion, or administration directly into a body cavity an outer exit duct, e.g. the gastrointestinal tract, bladder or uterus. So can the Compounds of the present invention in conventional pharmaceutical administration forms such as tablets, capsules, powders, solutions, suspensions, dispersions, Syrups, suppositories etc.

however become solutions, Suspensions and dispersions in physiologically acceptable carrier media, e.g. Water for Injections, generally preferred.

The Compounds according to the invention can therefore for the administration can be formulated using physiologically acceptable vehicle or aids in a way that is completely within the expertise lies. For example, you can the compounds, optionally with the addition of pharmaceutically acceptable Aids, in an aqueous Medium suspended or dissolved with the resulting solution or suspension then sterilized becomes.

For imaging of some parts of the body, the most preferred mode of administration of contrast agents is parenteral, eg, intravenous administration. Parenterally administrable forms, eg, intravenous solutions, should be sterile and free of physiologically unacceptable agents and should have low osmolality to minimize irritation or other adverse effects upon administration, and thus the contrast agent should preferably be isotonic or slightly hypertonic. Suitable vehicles include aqueous vehicles commonly used for administering parenteral solutions, such as sodium chloride injection, Ringer's Injection, Dextrose Injection, Dextrose and Sodium Chloride Injection, Lactated Ringer's Injection, and other solutions as described in Remington's Pharmaceutical Sciences, 15th Edition, Easton: Mack Publishing Co., pages 1405-1412 and 1461-1487 (1975) and The National Formulary XIV, 14th Edition Washington: American Pharmaceutical Association (1975). The solutions For example, preservatives, antimicrobials, buffers and antioxidants conventionally used for parenteral solutions may contain adjuvants and other additives that are compatible with the chelates and will not interfere with the manufacture, storage or use of the products.

The Means of the formula I can be therapeutically effective in the treatment of disease states as detectable in vivo imaging. So can e.g. the vector on the reporter units has a therapeutic effectiveness own, e.g. due to the radiotherapeutic effect of a Radionuclide repor ters, the efficacy in photodynamic therapy a chromophore (or fluorophore) reporter or chemotherapeutic Effects of the vector unit.

The Use of the agents of formula I in the preparation of therapeutic Compositions and in methods of therapeutic or prophylactic Treatment of the human or non-human animal body thus considered as a further aspect of the invention.

In an embodiment For example, the contrast agent of the invention conveniently comprises a gas-containing or gas-generating material, preferably in suspension in one aqueous support material and conjugated to one or more vectors, of which at least a vector V is as defined above, e.g. an AII receptor antagonist such as CV11974 or TCV-116. The gas may take the form of microbubbles passing through a Monolayer of a film-forming surfactant stabilized are stabilized or stabilized by a matrix material that is another is as a surface active Medium. The vectors can e.g. to such a surface-active Medium or a matrix and may be bioactive or non-bioactive. The Vectors can different targeting specificities and in a preferred embodiment they are such that they interact with their receptors but not the gas-containing ones Firmly bind vesicles.

The The present invention will now be further understood by means of the following restrictive Examples illustrated. Unless otherwise indicated, they are all given percentages Weight.

EXAMPLE 1

Angiotensin receptor binding Contrast agent for MR imaging

Connection 1

diethylenetriaminepentaacetic (53.8 g, 137 mmol) is dissolved in dry DMF (N, N-dimethylformamide) disbanded and 4 '- [3- (3-aminopropyl) -5-butyl-1H-1,2,4-triazol-1-yl) methyl] [1,1'-biphenyl] -2-carboxylic acid (prepared according to WO 91/17148, 5 g, 13.7 mmol) in dry DMF is added. The reaction mixture is at increased Temperature stirred under nitrogen atmosphere. The reaction is followed by TLC. The solvent is rotary evaporated and the substance is purified by chromatography.

Gd (III) chelate of the compound 1

To a solution of compound 1 (2 g, 2.7 mmol) in water is added gadolinium oxide Gd ₂ O ₃ (0.5 g, 1.4 mmol) and the mixture is heated at 95 ° C. After filtration, the solution is evaporated and dried in vacuo at 50 ° C.

EXAMPLE 2

Angiotensin receptor binding Contrast agent for nuclear medicine

Tc99m chelate of the compound 1

Compound 1 of Example 1 (1 mg) is dissolved in 0.1 N NaOH, SnCl ₂ 2H ₂ O (100 ug) dissolved in 0.05 N HCl and a solution of 10-100 mCi Tc-99m in the form of sodium pertechnetate in saline is added. The pH of the solution is adjusted to pH 7-8 by adding 0.5 M phosphate buffer (pH 5) in less than one minute. The reaction is followed by TLC and the substance is purified by chromatography.

EXAMPLE 3

Angiotensin receptor binding Contrast agent for nuclear medicine

An aqueous solution of ¹³¹ I ₂ (2 equivalents) and sodium perchlorate (1 equivalent) is added to an aqueous solution of 4 '- [3,5-dibutyl-1H-1,2,4-triazol-1-yl) methyl] [ 1,1'-biphenyl] -2-carboxylic acid (prepared according to WO 91/17148, (1 equivalent)). The solvent is rotary evaporated and the substance is purified by chromatography.

EXAMPLE 4

Angiotensin receptor binding Contrast agent for Ultrasound and light scattering imaging

Connection 2

succinic anhydride (2.7 g, 27.4 mmol) is dissolved in dry DMF and 4 '- (3- (3-aminopropyl) -5-butyl-1H-1,2,4-triazol-1-yl) methyl] 1,1'-biphenyl] -2-carboxylic acid (prepared according to WO 91/17148, 5 g, 13.7 mmol) in dry DMF is added. The reaction mixture is at increased Temperature stirred under nitrogen atmosphere. The reaction is followed by TLC. The solvent is rotary evaporated and the substance is purified by chromatography.

Connection 3

DSPE (Distearoylphosphatidylethanolamine) (0.5 g, 0.7 mmol) becomes a solution Compound 2 (0.3 g, 0.7 mmol) and DCC (N, N'-dicyclohexylcarbodiimide) in dry DMF given. The reaction mixture is at ambient temperature touched and the reaction is followed by TLC. The dispersion will stand overnight at 4 ° C calmly. The dispersion is filtered and the solvent rotavaporated before the substance is purified by chromatography becomes.

Gas-containing microparticles, the phosphatidylserine and the compound 3 included

To Phosphatidylserine (5 mg) is added 5% propylene glycol glycerol in water (1 ml). The dispersion is not more than 90 ° C for five minutes heated and then cooled to ambient temperature. The dispersion (0.8 ml) gets into a glass vial (1 ml) and compound 3 (0.1-1.0 mb) is added. The glass vial is on a scooter table for arranged for a few hours. The headspace of the glass vial is rinsed with perfluorobutane. The glass vial is used in a cap mixer for Shaken for 45 seconds, after which the sample is placed on a roller table. After centrifugation the lower fraction is exchanged with water. The washing procedure is repeated two to three times.

EXAMPLE 5

Tc-labeled angiotensin receptor binding Contrast agent for Scintigraphy: Tc chelate of 2- (4-N- (MAG-3) -aminophenyl) -1-benzyl-4-chloroimidazole-5-acetic acid hydrochloride

2-amino-3,3-dichlorpropenonitril (Connection)

A solution of dichloroacetonitrile (55.0 g, 0.5 mol) and acetone cyanohydrin (46.8 g, 0.55 mol) in acetonitrile (200 mL) and diethyl ether (50 mL) was cooled to 0 ° C. Potassium cyanide (10.65 g, 10.0 mmol) was added and the mixture was stirred under nitrogen atmosphere for 19 h at 0 ° C. The solution was evaporated, the brown residue was dissolved in diethyl ether (125 ml), treated with charcoal, filtered and evaporated. The residue was mixed with diethyl ether (30 ml) and hexanes (30 ml). The title compound, compound 4, was isolated by filtration.
Yield 21.2 g (pale yellow crystals)

The filtrate was evaporated, the residue was washed with ether / hexanes (1: 1, 20 ml). More of the title compound was isolated by filtration.
Total yield: 34.5 g (50%) mp 56-58 ° C.
NMR confirmed the structure of the title compound (~ 10% of cyanohydrin starting material as an impurity).

2- (4-nitrobenzylidene) amino-3,3-dichlorpropenonitril (Compound 5)

A solution of 2-amino-3,3-dichloropropenonitrile (15.0 g, 0.11 mol) and 4-nitrobenzaldehyde (15.1 g, 0.1 mmol) in toluene (200 mL) was refluxed for 23 hours , The mixture was evaporated, methanol (100 ml) added and the mixture allowed to stand overnight. After filtration, the title compound was isolated as a brownish yellow solid.
Yield 15.0 g, mp 171-174 ° C
Rf = 0.9 (silica, ethyl acetate: hexane = 1: 2)

Evaporation of the mother liquor gave 13.7 g of brown residue. Methanol (25 mL) was added and the mixture allowed to stand for two days. Filtration left 6.6 g dark brown material (title compound - compound 5).
Total yield: 21.6 g (80%)
NMR confirmed the structure of the title compound.

4-chloro-5-formyl-2- (4-nitrophenyl) imidazole (Compound 6)

HCl gas one left through a solution of 2- (4-nitrobenzylidene) amino-3,3-dichloropropenonitrile (40.0 g, 0.148 g mol) in dioxane (500 ml) at room temperature in bubbles. The mixture was stirred at 50 ° C for 23 hours. HCl gas was allowed to in Bubbling, followed by stirring for 24 hours at 50 ° C. The mixture was eight more days at 50 ° C stirred with addition of HCl gas every 24 hours, followed by stirring 50 ° C for four more Days. More HCl gas was added and stirred at 50 ° C for three more days. The solution was at 0 ° C chilled and filtered. The residue was mixed with ethanol (96%, 200 ml) and the title compound (Compound 6) was isolated by filtration. Yield: 23.1 g (62%). NMR confirmed the Structure of the title compound.

1-benzyl-4-chloro-5-formyl-2- (4-nitrophenyl) imidazole (Compound 7)

A Mixture of 4-chloro-5-formyl-2- (4-nitrophenyl) imidazole (19.7 g, 78.3 mmol), benzyl bromide (14.5 g, 85.0 mmol) and potassium carbonate (11.7 g, 85.0 mmol) in DMF (150 mL) was heated at 110 ° C for 1.5 h. The Mixture was evaporated and the residue poured into cold water and he was left overnight in the refrigerator stand. The tarry residue was pulverized and filtered. The yield of the crude title compound: 33.4 g. The product was recrystallized from ethanol (96%, 350 ml). Yield: 14.0 g. NMR confirmed the structure of the title compound, compound 7.

1-benzyl-4-chloro-5- (hydroxymethyl) -2- (4-nitrophenyl) imidazole (Compound 8)

Sodium borohydride (1.1 g, 29 mmol) was added in three portions to a stirred mixture of crude 1-benzyl-4-chloro-5-formyl-2- (4-nitrophenyl) imidazole (54.2 g, 88 mmol) in Methanol (250 ml) was added at room temperature. After two hours more sodium borohydride (0.51 g, 15 mmol) was added and the mixture was stirred for 30 minutes and the methanol was evaporated. The residue was mixed with water (250 ml) and extracted with methylene chloride (200 ml, 3 × 50 ml). The combined organic solutions were washed with water (2 x 50 ml) and dried overnight (NaSO ₄ ).

The mixture was filtered and evaporated, leaving a residue of 20.9 g. The title compound, compound 8, was purified by flash chromatography (150 x 65 mm silica gel 60 column eluted with ethyl acetate / hexanes (1: 1)). The fractions containing the title compound were evaporated.
Yield: 12.75 g (42%).
Rf = 0.4 (silica, ethyl acetate / hexanes (1: 1)).

1-benzyl-4-chloro-5- (chloromethyl) -2 (4-nitrophenyl) imidazole (Compound 9)

thionyl chloride (0.9 mL, 1.48 g, 12.4 mmol) was added dropwise to a stirred suspension of 1-benzyl-4-chloro-5- (hydroxymethyl) -2 (4-nitrophenyl) imidazole (18.5 g, 5.4 mmol) in chloroform (15 mL) at room temperature. The brown solution was stirred for 2.5 hours and then evaporated to dryness. Toluene (12 ml) became the residue given and evaporated to dryness. The residue was in chloroform (20 ml) dissolved and in the next Step to introduce the functional cyano group used.

1-benzyl-4-chloro-2 (4-nitrophenyl) -5-imidazolylacetonitrile (Compound 10)

A solution of sodium cyanide (1.60 g, 32.7 mmol) and tetrabutylammonium bromide (0.2 g) in ice-water (10 ml) was added dropwise to a vigorously stirred solution of 1-benzyl-4-chloro-5- (chloromethyl) Add 2- (4-nitrophenyl) -imidazole (5.4 mmol) in chloroform at 0 ° C. The mixture was stirred at 0 ° C for two hours and then allowed to warm to room temperature, diluted with water (10 ml), and the organic layer was washed with water (10 ml). The combined aqueous layers were extracted with chloroform (2 x 2 ml). The combined organic solutions were dried (Na ₂ SO ₄ ), filtered and evaporated, yielding 4.3 g. The product was purified by flash chromatography (170 x 25 mm, silica gel 60 column, eluted with ethyl acetate / hexanes 1: 1). Yield: 2.1 g.
Rf = 0.7 (silica, ethyl acetate / hexanes 1: 1)
NMR confirmed the structure of the title compound, Compound 10.

1-Benzyl-2- (4-nitrophenyl) -5-imidazole-acetic acid (Compound 11)

A stirred Mixture of 1-benzyl-4-chloro-2- (4-nitrophenyl) -5-imidazolylacetonitrile (2.0 g, 5.4 mmol) in 6 M HCl (20 mL) was allowed to stand for four hours backflow heated. The mixture was diluted with water (100 ml). A yellow oil fell out, that with cooling solidified to room temperature. Filtration gave 2.2 g of solid Material recrystallized from 96% ethanol (15 ml) and water (10 ml) has been.

A Filtration gave 2.24 g of wet residue. NMR confirmed the Structure of the title compound, compound 11. The product was obtained in next Step used.

2- (4-Aminophenyl) -1-benzyl-4-chloro-imidazole-5-acetic acid hydrochloride (Compound 12)

A mixture of 1-benzyl-2- (4-nitrophenyl) -5-imidazoleacetic acid (2.1 g, 5.0 mmol), 10% palladium on carbon (0.10 g), 12 M HCl (3.0 mL ), 96% ethanol (50 ml) and water (50 ml) was hydrogenated at 55 psi for 1.5 hours on a Parr shaker hydrogenation apparatus. The mixture was filtered through a celite pad and then through a fluted filter paper. Evaporation left the title compound, Compound 12, as 1.0 g of an orange solid.
Rf = 0.1 (silica, ethyl acetate / hexanes 1: 1)

Tc chelate of 2- (4-N- (MAG-3) -aminophenyl) -1-benzyl-4-chloro-imidazole-5-acetic acid hydrochloride (Compound 13)

The title compound, Compound 13, was prepared as set forth in the scheme above, as described in Nuclear Medicine Communications 16, (1995) 942-957.
Labeling yield:> 95%.

EXAMPLE 6

¹³¹ I-labeled 2- (4-aminophenyl) -1-benzyl-4-chloroimidazole-5-acetic acid hydrochloride

Compound 12 of Example 5 was ^labeled with ¹³¹ I using the chloramine T method as described in Radioisotopes in Biology, A Practical Approach. (1990) and in Nature 194, (162) 495.
Labeling yield: 56%

EXAMPLE 7

Tc-labeled angiotensin receptor binding contrast agent for Scintigraphy: Tc chelate of 2- (4-N- (MAG-3) -aminophenyl) -1-benzyl-4-chloro-5-hydroxymethylimidazole

2- (4-Aminophenyl) -1-benzyl-4-chloro-5- (hydroxymethylimidazolhydrochlorid (Compound 14)

A mixture of 1-benzyl-4-chloro-5- (hydroxymethyl) -2- (4-nitrophenyl) imidazole (1.0 g, 2.8 mmol, Compound 8 of Example 5), 10% palladium on carbon (0 10 g, 12 M HCl (1.0 mL) 96% ethanol (50 mL) and water (50 mL) was hydrogenated at 55 psi for 1.5 h on a Parr shaker hydrogenation apparatus And then filtered through a filter paper folded into a flute, leaving behind 1.5 g of amber-colored oil.
Rf = 0.2 (silica, ethyl acetate / hexanes 1: 1)

Tc chelate of 2- (4N- (MAG-3) -aminophenyl) -1-benzyl-4-chloro-5-hydroxymethylimidazole (Compound 15)

The title compound, compound 15, was prepared as described in Example 5, compound 13.
Labeling yield: 89%.

EXAMPLE 8

¹³¹ I-labeled N-benzyl losartan

N-benzyl losartan (Compound 16)

losartan (100 mg / 2.22 mmol) was dissolved in 3 ml of acetonitrile. benzyl bromide (38mg) was added and the reaction mixture became overnight stirred at ambient temperature. The solvent was evaporated in vacuo and the residue redissolved in chloroform, filtered and purified by column chromatography. The product was identified by MALDI mass spectrometry and used without further purification.

¹³¹ I-labeled N-benzyl losartan (Compound 17)

The Compound was made using an exchange tagging technique labeled as described in Radioisotopes in Biology, A practical Approach. (1990) and in Nature 194, (1962) 495. Labeling yield: 98%.

EXAMPLE 9

Tc-labeled losartan derivative for scintigraphy: Tc-chelate of N- (N-MAG-3-glycyl) -osartan

FmocGlycyl losartan (Compound 18)

Fmoc-Gly (300 mg / 1 mmol) was dissolved in 10 ml of THF. DCC (110 mg, 0.55 mmol) dissolved in 5 mL of THF was added and the reaction mixture was stirred overnight. DCC was filtered removed and the solvent removed in vacuo. TLC showed a UV + spot and the structure was confirmed by MALDI mass spectrometry.

The crude product was dissolved in 10 ml of THF and losartan 100 mg was added added. The stirring became overnight continued at ambient temperature. TLC showed a full conversion of the starting material. The solvent was removed in vacuo. The crude product was dissolved in chloroform and filtered through Purified by column chromatography. TLC showed a UV + spot, and the structure was determined by MALDI mass spectrometry approved.

Glycyl losartan (Compound 19)

100 mg FmocGlycyl losartan (compound 18) was dissolved in 5 ml of 10% piperidine / DMF disbanded and stirred for 30 minutes. TLC indicated full conversion of the UV + source material a new UV + ninhydrin + spot. The solvent was in vacuo evaporated. The structure was confirmed by MALDI mass spectrometry and the crude product was used without further purification.

Tc chelate of N- (N-MAG-3-glycyl) -osartan (Compound 20)

The title compound, compound 20, was prepared as described in Example 5, compound 13.
Labeling yield: 80%

EXAMPLE 10

Multi-specific, gas-containing microbubbles encapsulated with phosphatidylserine and biotin-PEG ₃₄₀₀ alanycholesterol and with streptavidin / biotinyl endothelin-1 peptide (biotin-D-Trp-Leu-Asp-Ile-Ile-Trp.OH) and Biotinyl-fibrin anti-polymerizing peptide (biotin-GPRPPERHQS.NH ₂ ) are functionalized

This Example is directed to the production of targeted ultrasonic microbubbles, wherein Streptavidin as a linker between biotinylated reporter (s) and vector (s) is used.

a) Synthesis of biotin-PEG ₃₄₀₀ -β-alanine cholesterol

To a solution of cholesteryl β-alanine hydrochloride (15 mg, 0.03 mmol) in 3 mL chloroform / wet methanol (2.6: 1) was added triethylamine (42 mL, 0.30 mmol). The mixture was stirred for 10 min at room temperature and a solution of biotin / PEG ₃₄₀₀ -NHS (100 mg, 0.03 mmol) in 1,4-dioxane (1 mL) was added dropwise. After stirring at room temperature for 3 h, the mixture was evaporated to dryness and the residue was purified by flash chromatography to give white crystals, yield 102 mg (89%). The structure was verified by MAL-DI-MS and NMR.

b) Synthesis of biotinylated Endothelin-1-peptide (biotin-D-Trp-Leu-Asp-Ile-Ile-Trp.OH).

The peptide was synthesized on an ABI 433 A automated peptide synthesizer starting from Fmoc-Trp (Boc) -Wang resin (Novabiochem) on a 0.1 mmol scale using 1 mmol amino acid cartridges. All amino acids were preactivated using HBTU before coupling. Concurrent removal of the peptide from the resin and side-chain protecting groups was carried out in TFA containing 5% anisole and 5% H ₂ O for 2 hours to give a crude product yield of 75 mg. Purification by preparative HPLC (Vydac 218TP1022 column) of an aliquot of 20 mg crude material was carried out using a gradient of 30 to 80% B over 40 min (A = 0.1 TFA / water and B = 0.1%. TFA (acetonitrile) and a flow rate of 9 ml / min After lyophilization of the pure fractions, 2 mg of pure material were obtained (analytical HPLC, gradient, 30-80% B, where B = 0.1% TFA / acetonitrile, A = 0.01% TFA / water: column - Vydac 218TP54; detection - UV 214 nm - product retention time = 12.6 min).

A further product characterization was performed using MALDI mass spectrometry; expects M + H at 1077, found 1077.

c) Synthesis of biotinyl fibrin anti-polymerizing peptide (biotin-GPRPPERHOS.NH ₂ )

This peptide was synthesized and purified using protocols similar to those are as described above in section b). The pure product was characterized by HPLC and MALDI-MS.

d) Preparation of multi-specific, gas-filled microbubbles encapsulated with phosphatidylserine and biotin-PEG ₃₄₀₀ -β-alanine cholesterol

DSPS (Avanti, 4.5 mg) and biotin-PEG ₃₄₀₀ -β-alanine cholesterol from section a) (0.5 mg) were weighed into a glass vial and 0.8 ml of a solution of 1.4% propylene glycol / 2.4% glycerol was added. The mixture was heated to 80 ° C for 5 minutes (the vials were shaken during heating).

The Sample was cooled to room temperature and the headspace became rinsed with perfluorobutane gas. The glass vials was in a cap mixer for Shaken for 45 sec and the microbubbles over Rolling and rolling the night. The microbubble suspension was deionized several times Washed water and with a Coulter counter and an acoustic damping analyzed.

e) Conjugation with fluorescein-labeled Streptavidin and biotinylated peptides from section b) and c)

To the Microbubble preparation of d) was fluorescein-conjugated streptavidin (0.2 mg) which was dissolved in PBS (1 ml). The microbubbles were placed on a roller table for 3 hours at room temperature. To detailed Washing with water and analysis by fluorescence microscopy were the microbubbles in 1 ml PBS, the biotinyl endothelin 1 peptide (0.5 mg) and biotinyl fibrin antipolymerizing peptide (0.5 mg) of the sections b) and c), incubated for 2 h. Expanded Washing of the microbubbles was carried out to unconjugated To remove peptide.

EXAMPLE 11

Multi-specific, filled with gas Microbubbles encapsulated with phosphatidylserine and biotinylated Lipopeptide used to make a streptavidin "sandwich" with a mixture Biotinyl Endothelin-1-Peptide (Biotin-D-Trp-Leu-Asp-Ile-Ile-Trp.OH) and biotinyl fibrin anti-polymerizing peptide (Biotin-GRPPERHQS.NH2)

a) Synthesis of lipopeptide Dipalmitoyl lysinyl -tryptophanyl-lysinyl-Iysinyl-lysinyl (biotin) glycine

The Lipopeptide was run on an automatic ABI 433A peptide synthesizer starting from Fmoc-Gly-Wang resin (Novabiochem) on a 0.1 mmol scale Use 1 mmol of amino acid cartridges synthesized. All amino acids and palmitic acid were preactivated using HBTU prior to coupling.

Concurrent removal of the peptide from the resin and side-chain protective chains was carried out in TFA containing 5% phenol, 5% EDT, 5% anisole, and 5% H ₂ O for 2 h, giving a crude product yield of 150 mg revealed. Purification by preparative HPLC (Vydac 218TP1022 column) of an aliquot of 40 mg of the crude material was carried out using a gradient of 70 to 100% B over 30 minutes (A = 0.1% TFA / water and B = MeOH) at a flow rate of 9 ml / min. After lyophilization, 14 mg of crude material (analytical HPLC; gradient, 70-100% B, where B = MeOH, A = 0.01 TFA / water: column - Vydac 218TP54: detection - UV 260 and fluorescence, Ex 280, Ein 350 - Product retention time = 22 minutes). Further product characterization was performed using MALDI mass spectrometry; expects M + H at 1478, found 1471.

b) Production of gas containing microbubbles on DSPS containing the biotinylated lipopeptide sequence of section a) are "doped"

DSPS (Avanti, 4.5 mg) and lipopeptide of a) (0.5 mg) were added to each of two glass bottles weighed and 0.8 ml of a solution of 1.4% propylene glycol / 2.4% glycerol was added to each glass vial given. The mixture was heated at 80 ° C for 5 minutes (the glass vials were during of heating ) Shaken. The samples were cooled to room temperature and the headspace became rinsed with perfluorobutane gas. The glass bottles were shaken in a cap mixer for 45 seconds and the microbubbles formed over Rolled night. The microbubbles were deionized several times Washed water and analyzed by a Coulter counter and acoustic attenuation.

MALDI mass spectral analysis was used to confirm incorporation into DSPS microbubbles.

c) Preparation of multi-specific, gas-filled microbubbles encapsulated with phosphatidylserine and a biotinylated lipopeptide and functionalized with streptravidin / biotinyl-endothelin-1-peptide (Biotin-D-Trp-Len-Asp-Ile-Ile-Trp.OH ) / Biotinyl fibrin anti-polymerizing peptide (biotin-GPRPPERHOS.NH ₂ )

The Microbubble preparation of b) above was in one of those in Example 10, section e) treated in an analogous way.

Claims (7)

Composition of the formula I VLR (I) wherein V is an organic group having a binding affinity for an angiotensin II receptor site and: [losartan], the radical of formula II,

wherein R ₁ and R _{2 are} carboxy groups or C _1-6 alkyl groups which are optionally substituted by oxo, oxo, phenyl, amino, carboxyl or hydroxy groups, or R ₁ additionally one Represent oxo group, and R ₃ and R _{4 are} methyl-biphenyl-carboxyl or methyl-biphenyl-tetrazole groups or C _1-6 -alkyl groups which are optionally substituted by oxo, oxo, phenyl, Amino, carboxyl or hydroxy groups are substituted, wherein R ₄ is present only when R _{1 is} oxo, wherein preferably one of the groups R ₃ and R _{4 represents} a biphenyl-containing group and one of the groups R ₁ , R ₂ , R ₃ and R _{4 represent} an alkyl group substituted with oxo, oxo, phenyl, amino, carboxyl or hydroxy groups,

wherein the double bond marked with an asterisk is optionally replaced by a single bond; R _{100 represents} a hydrogen atom, a phenyl group optionally substituted with at least one hydroxy or amino group, or a C _1-6 alkyl group optionally substituted with a hydroxy group or optionally an oxygen -, sulfur or nitrogen atom is bonded; R _{400 is} an arylmethyl group; R _{300 is} a carboxymethyl group or an optionally hydroxy-substituted C _1-6 alkyl group and R _{200 is} chlorine or R ₃₀₀ and R ₂₀₀ together with the interposed carbons form a condensed 6-membered carbocyclic or heterocyclic aromatic ring optionally substituted on a ring carbon with a hydroxy, carboxyl, C _1-6 alkyl or acyloxy group or on a ring nitrogen with an acyl group,

wherein R _{501 is} an optionally fluorinated C _1-6 alkyl group, R _{504 is} an optionally fluorinated C _1-6 alkyl group, R _{502 is} a carboxy, hydroxy-C _1-6 alkyl or CHO group stands; or the double bond marked with an asterisk is replaced by a single bond and R _{504 is} an = O or = NR ₅₀₅ group, R _{502 is} an optionally fluorinated C _1-6 alkyl group and R _{503 is} a carboxyphenyl group, and R _{30 is}

and R _{31 is} COOH, tetrazol-5-yl, NH ₂ SO ₂ CF ₃ , SO ₂ NHCO-phenyl, SO ₂ NHCO-cyclopropyl, L is a linker moiety or a bond, and R is a moiety which detectable in vivo imaging of a human or animal body. A composition of matter according to claim 1 selected from: i) a Tc chelate of 2- (4-N- (MAG-3) -aminophenyl) -1-benzyl-4-chloro-imidazole-5-acetic acid; ii) ¹³¹ I-labeled 2- (4-aminophenyl) -1-benzyl-4-chloro-imidazole-5-acetic acid; iii) a Tc chelate of 2- (4-N- (MAG-3) -aminophenyl) -1-benzyl-4-chloro-5-hydroxymethylimidazole; iv) ¹³¹ IN-benzyl-losartan; v) a Tc chelate of N- (N-MAG-3-glycyl) -losartan; vi) a Gd chelate of a conjugate of DTPA and 4 '- [3- (3-aminopropyl) -5-butyl-1H-1,2,4-triazol-1-yl) methyl] (1,1'-biphenyl Vii) a Tc chelate of a DTPA conjugate of 4 '- [3- (3-aminopropyl) -5-butyl-1H-1,2,4-triazol-1-yl) methyl] 1,1'-biphenyl] -2-carboxylic acid; viii) ¹³¹ 1-labeled 4 '- (3,5-dibutyl-1H-1,2,4-triazole-1-yl) methyl] [1,1'-biphenyl] -2-carboxylic acid; ix) a vesicle containing a succinic acid conjugate of 4 '- [3- (3-aminopropyl) -5-butyl-1H-1,2,4-triazol-1-yl) methyl] [1,1'-biphenyl ] -2-carboxylic acid carries. A composition of matter according to claim 1, characterized in that it is ¹²³ I-losartan. A composition of matter according to claim 1, characterized in that it is ¹³¹ I-losartan. Pharmaceutical composition with a composition of matter of the formula I according to any one of claims 1 to 4 together with at least a pharmaceutically active carrier or excipient. Use of a substance composition of the formula I according to one of the claims 1 to 4 for the preparation of a contrast medium for use in a diagnostic procedure involving the administration of the contrast medium to a living subject and the creation of an image at least of a part of the subject. Process for the preparation of a composition of matter of the formula I according to any one of claims 1 to 4, wherein the process comprises conjugating (i) an organic compound having binding affinity for one Angiotensin II receptor (ii) a compound used in a diagnostic imaging procedure is detectable, or a chelating compound, and, if necessary, Metalating the chelating groups in the resulting conjugate with a metal ion in a diagnostic imaging procedure is detectable.