DE69724817T2 - CONTRAST AGENTS - Google Patents

Description

This invention relates to diagnostic Imaging techniques in which a disease state under Imaged using a targeted contrast agent can, and targeted contrast agents which are for use in such techniques are suitable. More specifically, concerns Invention the use of such contrast media, the targeting or targeting vector binds to endothelin receptors.

The lining of the blood vessels, the Endothelium, secretes various means of relaxation and cause contraction of the vessels can, which regulates blood flow and pressure. Many cardiovascular diseases are associated with imbalances in the secretion of such agents, what z. B. leads to long-term structural damage.

The endotheline (ETs) are a family of related oligopeptides, which as a result of stimuli such as Hypoxia, shear stress, circulating hormones, growth factors and cytokines. The endothelins are vasoconstrictors, and indeed is the dominant isoform, ET-1, one of the most potent known Mammalian vasoconstrictors and is with different pathological effects in different Diseases and disorders associated. ET-1 is a 21-amino acid peptide, which vasoconstrictor, Has bronchoconstrictor and mitogenic activities and in many Cell types, e.g. B. neurons, glia cells, smooth vascular muscle, Endothelial and gastric mucosal cells is found.

The ability of ET-1 to control blood pressure to increase depends on its induction of vascular contraction via the Activation of different hormones together. However, this effect works a temporary vasodilator Effect ahead. ET-1 can also act as a mitogen to help proliferation of smooth muscle cells, and thus contributes to vascular proliferation disorders, like restenosis, at.

Two types of ET receptors have been identified, ET _A and ET _B. The ET _A receptors found on smooth muscle cells have high affinity for ET-1 and ET-2, while the ET _B receptors found in endothelial cells have affinity for all ET isoforms. ET _B receptor levels are higher than those of ETA receptors in organs such as the kidney and brain, while ETA receptors predominate in the heart and vascular system.

ET _A and ET _B receptors both contribute to vasoconstriction, while vasodilation appears to depend only on the ET _B receptors in endothelial cells.

In terms of the area of potentially dangerous Effects of the endotheline are various ET receptor antagonists have been developed, e.g. B. for use in improving the Hemodynamics (e.g. B. restoration of blood flow), reduction of inflammation, prevention of mitogenesis, etc. Thus, such antagonists are for use in a big one A variety of treatments have been proposed, e.g. B. of unstable and variant angina, asthma, arteriosclerosis, cerebral ischemia, myocardial ischemia, renal ischemia, Stroke, cerebral vascular spasm, congestive heart failure, vascular complications in diabetics, stomach cancer, gastric mucosal injury, hypertension (e.g. pulmonary hypertension), idiopathic pulmonary Fibrosis, migraines, Nephropathy, Reynaud's disease, subarachnoid hemorrhage and restenosis and other hyperproliferative vascular diseases.

It has now been found that it possible is, endothelin receptor sites in vivo using targeted Contrast agents in which the targeting vector has affinity for endothelin receptor owns. The endothelin receptors are generally within of the cardiovascular System and the gastrointestinal tract localized and are for such contrast media accessible, if administered in the bloodstream or gastrointestinal tract become. Hence, it is more targeted using such Contrast medium possible, Diseases and disorders, such as heart failure, atherosclerosis, restricted blood flow, gastrointestinal tract surface damage as well other vascular and gastric diseases and disorders, to be proven and also the progress of treatment for such diseases and disorders to monitor.

Viewed in one aspect, the invention therefore sees a composition of matter of the formula I. VLR (I) where V is a non-peptide organic group with binding affinity for an endothelin receptor site, L is a linker unit or a binding, and R is a unit that is detectable in in vivo imaging of a human or animal body.

If R is a radionuclide, is it is preferably an iodine, directly or indirectly bound to V, or it is a metal ion complexed by a chelating group in L, and / or it's from V through biodegradation of an organic Linker group L can be separated. (It should be noted that hydrogen radionuclides, although demonstrable in vitro, cannot meet the requirement that R in a is detectable in vivo imaging).

In many cases, the composition of matter or relevant composition of formula (I) be a characterizable connection. In others, it may be a combination of compounds that are bound or otherwise associated, e.g. B. are conjugated. For convenience, the composition in question will hereinafter be referred to as an agent.

From another point of view, The invention provides a pharmaceutical composition comprising an effective amount (e.g., an amount effective to enhance the Image contrast in in vivo imaging) of an agent of the formula I together with at least one pharmaceutically active carrier or Excipient.

Yet another aspect contemplated, the invention contemplates the use of an agent of formula I for the Manufacture of a contrast medium for use in a diagnostic procedure prior to the administration of the contrast medium to a living Subject and creating an image of at least a portion of the Subject includes.

Yet another aspect considered, the invention provides a method of forming an image a living human or non-human (preferably Mammal- or bird) animal subject, which previously had a contrast agent z. B. in the vascular system or the gastrointestinal tract was administered using the procedure creating an image of at least part of the subject, to which the contrast medium has been distributed, e.g. B. by Roentgen-, MR, ultrasound, scintigraphy, PET, SPECT, electrical impedance, Includes light or magnetometry imaging modalities characterized that as the contrast medium an agent of formula I is used.

Yet another aspect considered, the invention provides a method of making an agent of formula I, wherein the method is conjugating (i) one organic non-peptide compound with binding affinity for one Endothelin receptor at (ii) a compound which is used in a diagnostic Imaging process is detectable, or a chelating compound and, if necessary, metallizing chelating groups in the resulting conjugate with a metal ion, which in a diagnostic Imaging process is detectable.

Have the agents of formula (I) three characteristic components: a vector (V); a linker (L); and a reporter (R). The vector must have the ability to connect to direct endothelin receptors, the reporter must be in one be detectable in vivo diagnostic imaging processes; and the linker must Couple the vector to the reporter, at least until the imaging process has been completed.

vectors

Any non-peptide can be used as a vector Connection with affinity use for endothelin receptors.

Non-peptide compounds will be used because peptide vectors are generally low biological stability exhibit and undesirable ET-like responses on the part of the body. With the term non-peptide excludes compounds which have a -CHR-CO-NH-CHR-CO-NH unit. Furthermore the vector may be a compound (such as BMS-182874) which is a more pronounced affinity for one Type of edothelin receptor than for the other type, or a compound (like bosentan) which affinity for both Possesses types of endothelin receptors. Connections with a more pronounced affinity for one particular type of endothelin receptors are generally preferred become.

The agent is preferably one Compound that does not produce an unacceptable biological response, especially compounds that act and do not act as endothelin receptor antagonists evoke the responses associated with endothelin itself are, especially the blood pressure modifying and mitogenic responses. However, you can biological responses, if desired by administration of a therapeutic agent e.g. B. before modified at the same time or after administration of the agent of formula I.

Among the non-peptide vectors become Bosentan, Shionogi's 97-139, Shionogi's 50-235, BMS-182874, Merck's L-744453, PD-155719, PD-155080, PD-156707, Fujisawa's FR 901366, FR 901367, PD-160672, PD-160874, SB 209670, SB 217242, L-749329, L-751281, IRL 2500, IRL 2659, IRL 2796, RO 468443, RO 46-2005, RO 470203, BMS 193884, TBC 11251, CGS-26303, CGS-26393, CGS-27830, L-751281, L-747844 and L-754142 prefers.

Examples of suitable endothelin receptor antagonists are specified in WO-95/03295, US-A-5420123, WO-94/27979, WO-95/03044 and EP-A-405421 and by Sakurawi et al. in Chem. Pharm. Bull. 44: 343-351 (1996). The development of suitable non-peptide endothelin receptor antagonists will discussed for example by Ferro et al. in Drugs 51: 12-27 (1996), Battistini et al. in TiPS 16: 217-222 (1995), and Doherty, Chapter 4, in "Chemical and Structural Approaches to Rational Drug Design ", ed .: Weiner et al., CRC Press, 1995.

Suitable non-peptide vectors will generally be polycyclic compounds holding a phenyl group linked to a 5- or 6-membered carbocyclic or heterocyclic ring by one or two bridging groups, one of which contains or is pending on a heteroatom-interrupted, oxo-substituted unit (e.g. a -SO ₂ -NH- or -CO-S- or -CO-CH ₂ -O group).

Thus the vector compound can be, for example, a compound of formula II ϕ – Bd – Rg (II) wherein ϕ is an optionally substituted phenyl group, optionally part of a fused polycyclic structure with up to 4 fused rings; Bd is a 1 to 5 atom long, optionally unsaturated organic bridge, with backbone atoms selected from C, N, S and O, optionally equipped with side chain substituents and optionally forms part of a condensed ring, and containing one

Unit in which X is C or SO, n Is 0 or 1 and Y is O, S or an amino nitrogen atom; and Rg a five- or six-membered, more saturated or unsaturated, optionally substituted carbocyclic or heterocyclic ring is optionally part of a condensed polycyclic structure with up to 5 condensed rings; the optional substituents selected are made of halogen and oxygen atoms, carbon and sulfur oxy acid groups, Hydroxy, alkyl, aryl, aralkyl, acyl, alkoxy, alkylenedioxy, Alkylthio, alkylamino, acyloxy, aralkylamino, aralkyloxy, acylthio, Amino and acylamino groups and combinations thereof (e.g. haloalkyl), and wherein the ring heteroatoms are selected from O, N and S, aryl groups mono- or bicyclic, carbocyclic or heterocyclic rings with up to 10 ring atoms, and no more than 7 rings and not there are more than 6 condensed rings.

In the compounds of formula II contain alkyl or alkylene groups preferably 1 to 6 carbon atoms, Aryl units are preferably phenyl or naphthyl groups or 5-membered heterocycles, which optionally have a condensed benzene or bear a 6-membered aromatic heterocyclic ring, and Halogen atoms are preferably F, Cl, Br or I.

In such connections, the Amino, hydroxy, carbonyl or oxyacid functions are easily used to conjugate the vector to the reporter.

ein, worin R¹ und R², die gleich oder unterschiedlich sein können, Hydroxy-, Schwefeloxysäure-, Acylamino- oder Carboxyacylaminogruppen sind (worin die Acyleinheit zweckmäßigerweise 1 bis 6 Kohlenstoffatome enthält).Thus, examples of such polycyclic vectors include triterpenoids of Formula III:

one wherein R ¹ and R ² , which may be the same or different, are hydroxy, sulfuroxyacid, acylamino or carboxyacylamino groups (wherein the acyl moiety conveniently contains 1 to 6 carbon atoms).

und

aufweist.Examples of such tripernoids include compounds in which the pendant phenyl group has the formula:

and

having.

In these compounds the hydroxyl and / or OSO ₃ H groups on the pending phenyl groups can expediently be used for the vector attachment.

(worin jedes R³, das gleich oder unterschiedlich sein, Wasserstoff oder eine Acylgruppe ist; R⁴ eine Carboxy- oder Acylgruppe ist; und R⁵ Wasserstoff, Hydroxy- oder eine Acyloxygruppe ist, wobei die Acylgruppen beispielsweise C_1-6-Gruppen, wie Formyl, Acetyl oder Propionyl, sind).Further examples of polycyclic vectors of formula II include anthraquinone compounds of formula IV:

(wherein each R ³ , the same or different, is hydrogen or an acyl group; R ^{4 is} a carboxy or acyl group; and R ^{5 is} hydrogen, hydroxy or an acyloxy group, the acyl groups being, for example, C _1-6 groups, such as formyl, acetyl or propionyl).

In these connections is the bridging group Bg part of the condensed tetracyclic structure.

Examples of vectors of formula IV include compounds wherein R ³ on the tetracyclic ring structure is all hydrogen, the pendant alkylthio group is 2-carboxy-2-acetylamino-ethylthio and R ^{5 is} hydrogen (compound FR 901366) or hydroxy (FR 901367) is.

In these connections, the Amino and / or carbonyl functions of the pendant alkylthio groups expediently for one Vector tack can be used.

(worin R⁶ Wasserstoff oder Alkyl ist, R⁷ Aryl, Aralkyl, Arylcarbonyloxy oder Aralkylcarbonyl ist (z. B. worin die Aryleinheit eine Phenyl-, Naphthyl- oder Indolyl-, z. B. Indol-3-yl-Gruppe ist, und jede Alkyleinheit 1 bis 6 Kohlenstoffatome besitzen kann); Z eine wahlweise Oxa-substituierte C_1-6-Alkylengruppe ist, bei der wahlweise eine CH₂-Einheit ersetzt ist durch ein Aminstickstoff- oder ein Sauerstoffatom (z. B. eine -CH₂-, -OCH₂-,

Z¹⁰ Wasserstoff oder eine Gruppe X₁, wie definiert in US-A-5420123, ist; und R⁸ Wasserstoff oder eine wahlweise substituierte C_1-6-Alkyl; z. B. Carboxyalkyl-, Aminoalkyl-, Sulfonyloxyalkyl-Gruppe etc. ist).Yet other examples of the polycyclic vectors include dibenzodiazepines of Formula V:

(where R ^{6 is} hydrogen or alkyl, R ^{7 is} aryl, aralkyl, arylcarbonyloxy or aralkylcarbonyl (e.g. where the aryl moiety is phenyl, naphthyl or indolyl, e.g. indol-3-yl group, and each alkyl unit can have 1 to 6 carbon atoms); Z is an optionally oxa-substituted C _1-6 alkylene group in which a CH ₂ unit is optionally replaced by an amine nitrogen or an oxygen atom (e.g. a -CH ₂ -, -OCH ₂ -,

Z ^{10 is} hydrogen or a group X ₁ as defined in US-A-5420123; and R ^{8 is} hydrogen or an optionally substituted C _1-6 alkyl; z. B. carboxyalkyl, aminoalkyl, sulfonyloxyalkyl group etc.).

Examples of R ⁷ groups thus include phenyl, naphthyl, indol-3-yl, benzyl, naphthylmethyl, indole-2-carboxyl, indole-3-carboxyl and 3-indolylacetyl. Examples of these dibenzodiazepines of formula V include compounds wherein R ^{6 is} hydrogen or propyl, R ^{8 is} carboxymethyl, and ZR ⁷ indole-3yl-methyl, indole (2 or 3) ylcarbonyloxy, indole (2 or 3) ylcarbonylamino, indole- 3-yl-methylcarbonyloxy, indol-3-ylmethylcarbonylamino, 1-naphthyl-methylamino, 1-naphthylmethoxy, benzyloxy and benzylamino.

With such compounds, the R ⁸ group can conveniently be used for vector attachment.

ein, (worin die Isoxazoylgruppe an den 5- oder 3-Positionen angebracht ist; R⁹ ein gegebenenfalls substituierter, gegebenenfalls kondensierter Benzolring (z. B. Amin- oder Oxysäure-substituiert), ein kondensierter Pyridinring, eine Phenylgruppe oder eine Carboxyl- oder Aminogruppe ist, wobei der Ring, an den sie gebunden ist, ungesättigt ist, oder eine kondensierte 2,2-Propyliden-Gruppe oder ein Sauerstoffatom ist, wobei der Ring, an den sie gebunden ist, gesättigt ist; und R¹⁰ eine geradkettige oder verzweigte C_1-6 Alkylgruppe oder ein Halogenatom ist).Still further examples of polycyclic vectors of formula I include N-isoxazolylsulfonamides, especially sulfonamides of formula VI:

one (where the isoxazoyl group is attached at the 5- or 3-positions; R ^{9 is} an optionally substituted, optionally fused benzene ring (e.g. substituted amine or oxyacid), a fused pyridine ring, a phenyl group or a carboxyl or Amino group, the ring to which it is attached is unsaturated, or a fused 2,2-propylidene group or an oxygen atom, the ring to which it is attached is saturated, and R ^{10 is} a straight chain or is branched C _1-6 alkyl group or a halogen atom).

ein. Bei derartigen Verbindungen kann die Vektoranheftung zweckmäßigerweise unter Verwendung von aktiven Substituenten in R¹⁰ oder R⁹, z. B. Amin- oder Oxysäuregruppen, bewirkt werden.Examples of vectors of the formula VI include compounds of the formulas:

on. In such compounds, vector attachment may conveniently be carried out using active substituents in R ¹⁰ or R ⁹ , e.g. B. amine or oxyacid groups.

(worin R¹¹ Wasserstoff oder C_1-6-Alkyl ist, wahlweise substituiert durch Oxysäure- oder andere reaktive Substituenten zur Vektorverknüpfung (z. B. Carboxyl); R¹² C_1-6-Alkyl ist, wahlweise substituiert durch Oxysäure oder andere reaktive Substituenten für die Vektorverknüpfung; R¹³ Wasserstoff, Halogen oder C_1-6-Allcyl ist oder eine Gruppe R¹⁰ wie definiert in der WO 95/03044; und R¹⁴ Wasserstoff, C_1-6-Alkoxy oder Methylendioxy oder eine Gruppe R¹, R², R^3a R^3b oder R¹⁰ wie definiert in WO 95/03044 ist).Further examples of polycyclic vectors of formula II include phenoxyphenylacetic acid substituted benzimidazolinones of formula VII.

(wherein R ^{11 is} hydrogen or C _1-6 alkyl optionally substituted by oxyacid or other reactive vector linking substituents (e.g. carboxyl); R ^{12 is} C _1-6 alkyl optionally substituted by oxy acid or other reactive Substituents for the vector linkage; R ^{13 is} hydrogen, halogen or C _1-6 alkyl or a group R ¹⁰ as defined in WO 95/03044; and R ^{14 is} hydrogen, C _1-6 alkoxy or methylenedioxy or a group R ¹ , R ² , R ^3a R ^3b or R ^{10 is} as defined in WO 95/03044).

Thus examples of vectors close Formula VII the following:

The vector attachment of such connections on linker reporter units using the carboxyl or other reactive substituents can be achieved.

ein (worin Z'' ausgewählt ist aus Alkylsulfonylaminocarbonyl, Arylsulfonylaminocarbonyl, Aryl, Arylaminocarbonyl und Carboxy; R¹⁵ Wasserstoff, Alkyl, Oxysäure oder eine andere reaktive Gruppe für die Vektorverknüpfung ist; R¹⁶ Wasserstoff, Halogen, Carboxyl, C_1-6-Alkoxy-carbonyl oder C_1-6-Alkyl, Alkoxyl, Hydroxy-alkyl oder Halogenalkyl ist; und R¹⁷ Wasserstoff, Halogen, Nitro, Carboxyl. Aminocarbonyl, Acetylamino, Alkoxycarbonylamino, Alkoxy oder Alkylendioxy ist). Falls Z'' eine Aryl-Funktion enthält, kann diese carbocyclisch (z. B. Phenyl) oder heterocyclisch (z. B. Tetrazolyl) sein und kann gegebenenfalls substituiert sein, z. B. durch Halogenatome, Carboxygruppen oder C_1-6-Alkyl- oder Halogenalkylgruppen.Still further examples of polycyclic vector compounds of formula II include 1,2-diphenylethylene compounds of formula I described in WO 95/03295, e.g. B. Compounds of Formula VIII:

a (wherein Z '' is selected from alkylsulfonylaminocarbonyl, arylsulfonylaminocarbonyl, aryl, arylaminocarbonyl and carboxy; R ^{15 is} hydrogen, alkyl, oxyacid or another reactive group for vector linking; R ^{16 is} hydrogen, halogen, carboxyl, C _1-6 alkoxy carbonyl or C _1-6 alkyl, alkoxyl, hydroxyalkyl or haloalkyl; and R ^{17 is} hydrogen, halogen, nitro, carboxyl, aminocarbonyl, acetylamino, alkoxycarbonylamino, alkoxy or alkylenedioxy). If Z ″ contains an aryl function, this may be carbocyclic (e.g. phenyl) or heterocyclic (e.g. tetrazolyl) and may optionally be substituted, e.g. B. by halogen atoms, carboxy groups or C _1-6 alkyl or haloalkyl groups.

ein (worin jedes R¹⁵¹ unabhängig Wasserstoff, Halogen oder Carboxy oder C_1-6-Alkyl, Alkoxy, Hydroxyalkyl oder Halogenalkyl, oder Alkoxycarbonyl, oder C_3-7-Cycloalkyl ist; R¹⁶ Wasserstoff oder C_1-6-Alkyl ist; und jedes R¹⁷¹ unabhängig Wasserstoff, Halogen, Nitro, Alkyl, Alkoxy, Aminocarbonyl, Alkoxycarbonyl, Alkoxycarbonylamino oder Acetylamino ist).Examples of the vector compounds of Formula VIII include:

a (wherein each R ^{151 is} independently hydrogen, halogen or carboxy or C _1-6 alkyl, alkoxy, hydroxyalkyl or haloalkyl, or alkoxycarbonyl, or C _3-7 cycloalkyl; R ^{16 is} hydrogen or C _1-6 alkyl; and each R ^{171 is} independently hydrogen, halogen, nitro, alkyl, alkoxy, aminocarbonyl, alkoxycarbonyl, alkoxycarbonylamino or acetylamino).

(worin jedes R¹⁵¹ unabhängig Halogen, n-Propyl oder i-Butyl ist; Z'' 4-i-Propyl-phenyl-SO₂NHCO ist; R¹⁶ Wasserstoff oder Methyl ist; und jedes R¹⁷¹ unabhängig Wasserstoff, Chlor oder Methoxy ist),

(worin R¹⁵¹ Wasserstoff, Carboxyl oder Hydroxymethyl ist; jedes R151 unabhängig Wasserstoff oder n-Propyl ist; Z'' Carboxy, 4-i-Propyl-phenyl-SO₂NHCO, 4-t-Butyl-phenyl-SO₂NHCO, Tetrazol-5-yl oder Tetrazol-5-yl-NHCO ist; R16 Wasserstoff oder Methyl ist; und R¹⁷¹ Wasserstoff, Chlor oder Methoxy ist),

(worin R¹⁵² Carboxy oder Hydroxymethyl ist; jedes R¹⁵¹ unabhängig Wasserstoff oder n-Propyl ist; Z" Carboxy, 4-i-Propyl-phenyl-SO₂NHCO, 4-i-Butyl-phenyl-SO₂NHCO, 4-Brom-phenyl-SO₂NHCO, Tetrazol-5-yl, Tetrazol-5-yl-NHCO oder i-Propyl-SO₂-NHCO ist; und R¹⁶ Wasserstoff oder Methyl ist) und

(worin R¹⁵² Carboxyl oder Hydroxymethyl ist; R¹⁵¹ Wasserstoff oder n-Propyl ist; R¹⁶ Wasserstoff oder Methyl ist; und Z'' Carboxy, 4-i-Propylphenyl-SO₂NHCO, 4-i-Butylphenyl-SO₂NHCO, 4-Brom-phenyl-SO₂NHCO, Tetrazol-5-yl, Tetrazol-5-yl-NH-CO oder i-Propyl-SO₂-NHCO ist; und R¹⁶ Wasserstoff oder Methyl ist) ein.For example, such compounds include compounds of the formulas:

(wherein each R ^{151 is} independently halogen, n-propyl or i-butyl; Z " 4-i-propyl-phenyl-SO ₂ NHCO; R ^{16 is} hydrogen or methyl; and each R ^{171 is} independently hydrogen, chlorine or methoxy is)

(wherein R ^{151 is} hydrogen, carboxyl or hydroxymethyl; each R151 is independently hydrogen or n-propyl; Z '' carboxy, 4-i-propyl-phenyl-SO ₂ NHCO, 4-t-butyl-phenyl-SO ₂ NHCO, Tetrazol-5-yl or tetrazol-5-yl-NHCO; R16 is hydrogen or methyl; and R ^{171 is} hydrogen, chlorine or methoxy),

(wherein R ^{152 is} carboxy or hydroxymethyl; each R ^{151 is} independently hydrogen or n-propyl; Z "carboxy, 4-i-propyl-phenyl-SO ₂ NHCO, 4-i-butyl-phenyl-SO ₂ NHCO, 4- Bromophenyl-SO ₂ NHCO, tetrazol-5-yl, tetrazol-5-yl-NHCO or i-propyl-SO ₂ -NHCO; and R ^{16 is} hydrogen or methyl) and

(wherein R ^{152 is} carboxyl or hydroxymethyl; R ^{151 is} hydrogen or n-propyl; R ^{16 is} hydrogen or methyl; and Z " carboxy, 4-i-propylphenyl-SO ₂ NHCO, 4-i-butylphenyl-SO ₂ NHCO , 4-bromophenyl-SO ₂ NHCO, tetrazol-5-yl, tetrazol-5-yl-NH-CO or i-propyl-SO ₂ -NHCO; and R ^{16 is} hydrogen or methyl).

(worin jedes R¹⁸, das unterschiedlich oder gleich sein kann, eine Alkoxy- oder Alkylendioxy-Gruppe ist), z. B. die Verbindungen:Further examples of polycyclic vectors of formula II include 3-benzoyl-2-phenyl-3- (benzyl or cyclohexylmethyl) propenoic acids and the furan analogs of formulas IX and X:

(wherein each R ¹⁸ , which may be different or the same, is an alkoxy or alkylenedioxy group), e.g. B. the connections:

ein (worin jedes R¹⁸, das gleich oder unterschiedlich sein kann, eine Alkoxy- oder Alkyldioxy-Gruppe ist), z. B. die Verbindungen:Still further examples of polycyclic vector compounds of formula II include compounds of formula XI:

one (wherein each R ¹⁸ , which may be the same or different, is an alkoxy or alkyl dioxy group), e.g. B. the connections:

ein (worin R¹⁹ C_1-6-Alkyl ist; R¹⁸ wie obenstehend definiert ist; Z''' O oder CH₂ ist; und R²⁰ Alkoxyaryloxy, Hydroxyalkoxy oder Alkoxyaryl ist), z. B. Verbindungen der Formeln:Still further examples of vector compounds of formula II include phenylsulfonamides of formulas XII and XIII:

a (wherein R ^{19 is} C _1-6 alkyl; R ^{18 is} as defined above; Z '''is O or CH ₂ ; and R ^{20 is} alkoxyaryloxy, hydroxyalkoxy or alkoxyaryl), e.g. B. Connections of the formulas:

ein (worin R¹⁹ wie obenstehend definiert ist; R²¹ eine 5- oder 6-ringgliedrige Arylgruppe ist, z. B. eine Phenyl-, Thiophenyl oder Isoxazolylgruppe; und jedes R²², das gleich oder unterschiedlich sein kann, Wasserstoff oder eine C_1-6-Alkylgruppe ist), z. B. Verbindungen der Formeln:Still further examples of the polycyclic vectors of Formula II include the compounds of Formula XIV

a (wherein R ^{19 is} as defined above; R ^{21 is} a 5- or 6-membered aryl group, e.g., a phenyl, thiophenyl or isoxazolyl group; and each R ²² , which may be the same or different, is hydrogen or a C. _1-6 alkyl group), e.g. B. Connections of the formulas:

ein (worin R¹⁸ wie obenstehend definiert ist; R²³ eine Aralkylgruppe, z. B. eine 3,6-Diazaindol-1-yl-methyl-Gruppe ist; und R²⁴ ein Aralkyl ist), z. B. eine Verbindung der FormelStill further examples of polycyclic vector compounds of Formula II include the phenoxy-phenylacetic acids of Formula XV:

a (wherein R ^{18 is} as defined above; R ^{23 is} an aralkyl group, e.g., 3,6-diazaindol-1-yl-methyl group; and R ^{24 is} an aralkyl), e.g. B. a compound of formula

ein (worin R²⁵ eine Oxysäure oder ein Ester davon ist, z. B. eine Alkoxycarbonylgruppe; R¹⁸ wie obenstehend definiert ist; und R²⁶ ein Wasserstoffatom oder eine wahlweise substituierte C_1-6-Alkylgruppe ist, welche z. B. eine reaktive funktionelle Gruppe trägt, die fähig zur Verwendung für die Vektorverknüpfung ist), z. B. eine Verbindung der Formel:Yet further examples of formula I polycyclic vectors include compounds of formula XVI

a (wherein R ^{25 is} an oxyacid or an ester thereof, e.g. an alkoxycarbonyl group; R ^{18 is} as defined above; and R ^{26 is} a hydrogen atom or an optionally substituted C _1-6 alkyl group, e.g. reactive functional group capable of use for vector linkage), e.g. B. a compound of the formula:

Other compounds used suitable as vectors include compounds such as those which in WO 95/03295, WO 95/03044, WO 94/27979, US-A-5420123, EP-A-405421 and WO 95/08550, and by Schin-ichi et al., in Eur. J. Pharmacol.-Mol. Pharmacol. 247: 219-221 (1993), and by Sakurawi et al. in Chem. Pharm. Bull. 44: 343-351 (1996), have been described.

The vectors of the formulas II to XVI can by techniques described in the patent and others Publications, to which reference is made herein.

Also CAM-D and other candidate identification and evaluation techniques, as mentioned above, can be applied to further Candidate non-peptide vectors to find or check.

So it is also possible to use molecules that bind specifically to endothelin receptors by direct screening of molecule libraries to obtain. Screening peptide libraries can also be used to identify generally effective peptide structures of which non-peptide analogs by conventional or combinatorial Chemistry can be generated. Binding units identified in this way can to a linker molecule be coupled, which is a general tool for connecting a any vector molecule (or molecules) to the reporter.

Vector molecules can be obtained from combinatorial libraries are produced without necessarily having the exact molecular Know the goal by working functionally (in vitro, ex vivo or in vivo) in terms of molecules selects which of the region / structure to be mapped, tie.

As mentioned above, the Formula I agents include vector, linker and reporter units. A linker unit can serve to link a vector to a reporter; alternatively, it can connect more than one vector and / or more than one reporter. In the same way, a reporter or a vector can be linked to more than one linker. Such use of a plurality of reporters (e.g., multiple linker-reporter units linked to a vector, or multiple reporters linked to a linker, which in turn is linked to a vector) can allow the detectability of the contrast agent to be increased (e.g., by increasing its radio capacity, echogenicity or relaxivity), or may allow it to be detected in more than one imaging modality. Such use of a plurality of vectors may increase the targeting efficiency of the contrast agent or may render the contrast agent capable of targeting more than one site, e.g. B. different receptors for an agent which has a receptor heterogeneity. For example, the agent of Formula I may include vector units with affinity sites other than endothelin receptors, e.g. B. with affinities for cell surfaces on wall surfaces of body ducts. Thus, the agent can use vectors such as antibody fragments and oligopeptides, e.g. B. containing RGD or analog cell surface binding peptide motifs (e.g. as described in EP-A-422937 and EP-A-422938 (Merck)), or other vectors as described in GB 9700699.3. Such extravectors can also be chosen from any of the molecules that naturally concentrate in vivo in a selected target organ, tissue, cell or cell group or other location in a mammalian body. These can include amino acids, oligopeptides (e.g. hexapeptides), molecular recognition units (MRU's), single chain antibodies (SCA's), proteins, organic non-peptide molecules, Fab fragments and antibodies. Examples of site-directed molecules include polysaccharides (e.g. CCK and hexapeptides), proteins (such as lectins, asialofetuin, polyclonal IgG, blood coagulation proteins (e.g. hirudin), lipoproteins and glycoproteins), hormones, Growth factors, coagulation factors (such as PF4), polymerized Firbrin fragments (e.g. E ₁ ), serum amyloid precursors (SAP) proteins, low-density lipoprotein (LDL) precursors, serum albumin, surface proteins from 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 Include fibrinogen. The site-directed protein can also be an antibody. The selection of the antibody, in particular the antigen specificity of the antibody, will depend on the target site specifically intended for the agent. Monoclonal antibodies are preferred over polyclonal antibodies. The 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 is particularly preferred that such Extravectors should bind so that the movement of the agent slows down in the bloodstream but does not prevent it and puts it in place and site is anchored when it is at an endothelin receptor site is bound.

left

The linker component of the contrast medium is in its simplest form a bond between the vector and the reporter unit. But the linker becomes even more general provide a mono- or multimolecular skeleton that is covalent or non-covalent linked one or more vectors to one or more reporters, e.g. B. a linear, cyclic, branched or reticular molecular Skeleton or a molecular aggregate with built-in or attached groups, which covalent or non-covalent, z. B. coordinate, bind with the vector and reporter units or which encapsulate, include, or such units anchor.

Therefore, linking a Reporter unit to a desired one Vector by covalent or non-covalent means, usually including the interaction with one or more functional groups, which are located on the reporter and / or vector become. Examples of chemically reactive functional groups, which for this purpose can be used conclude Amino, hydroxyl, sulfhydryl, carboxyl and carbonyl groups as well Carbohydrate groups, vicinal diols, thioethers, 2-amino alcohols, 2-aminothiols, guanidinyl, imidazolyl and phenolic groups on.

Covalent coupling of the reporter and vector can therefore be effected using linkers which contain reactive units which are capable of reacting with such functional groups. Examples of reactive units capable of reacting with sulfhydryl groups include α-haloacetyl compounds of the type X-CH ₂ CO- (where X = Br, Cl or I), which show particular reactivity for sulfhydryl groups, but which 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 to be selective for sulfhydryl groups, but may additionally be useful in coupling to amino groups under certain conditions. Reagents such as 2-iminothiolane e.g. B. as described by Traut, R., et al., In Biochemistry (1973) 12, 3266, which introduce a thiol group by converting an amino group, can be regarded as sulfhydryl reagents if the linkage takes place via the formation of disulfide bridges. Thus, reagents which introduce reactive disulfide bonds into either the reporter or the vector can be useful since linkage can be established by disulfide exchange between the vector and the reporter; Examples of such reagents include Ellman's reagent (DTNB), 4,4'-dithiodipyridine, methyl 3-nitro-2-pyridyl disulfide and methyl 2-pyridyl disulfide (described by Kimura, T., et al. In Analyt. Biochem. (1982) 122, 271).

• i) α-Halogenacetyl-Verbindungen, welche Spezifität gegen Aminogruppen in Abwesenheit von reaktiven Thiolgruppen zeigen und von dem Typ X-H₂CO- (worin X = Cl, Br oder I) sind, z. B. wie beschrieben von Wong, Y.-H.H. in Biochemistry (1979), 24, 5337;
• ii) N-Maleimid-Derivate, welche mit Aminogruppen entweder durch eine Reaktion vom Michael-Typ oder durch Acylierung durch Addition an die Ring-Carbonylgruppe, wie beschrieben von Smyth, D. G. et al., in J. Am. Chem. Soc. (1960) 82, 4600, und Biochem. J. (1964) 91, 589, reagieren können;
• iii) Arylhalogenide, wie reaktive nitrohalogenaromatische Verbindungen;
• iv) Alkylhalogenide, wie beschrieben von McKenzie, J. A. et al., in J. Protein Chem. (1988) 7, 581;
• v) Aldehyde und Ketone, fähig zur Schiff-Basen-Bildung mit Aminogruppen, wobei die gebildeten Addukte üblicherweise durch Reduktion unter Erhalt eines stabilen Amins stabilisiert werden;
• vi) Epoxid-Derivate, wie Epichlorhydrin und Bisoxirane, welche mit Amino-, Sulfhydryl- oder phenolischen Hydroxylgruppen reagieren können;
• Vii) Chlorhaltige Derivate von s-Triazinen, welche sehr reaktiv gegenüber Nukleophilen, wie Amino-, Sulfhydryl- und Hydroxygruppen, sind;
• viii) Aziridine, basierend auf den obenstehend ausführlich angegebenen s-Triazin-Verbindungen, z. B. wie beschrieben von Ross, W. C. J. in Adv. Cancer Res. (1954) 2, 1, welche mit Nukleophilen wie Aminogruppen durch Ringöffnung reagieren;
• ix) Quadratsäurediethylester, wie beschrieben von Tietze, L. F. in Chem. Ber. (1991) 124, 1215; und
• x) α-Halogenalkylether, welche reaktivere Alkylierungsmittel als normale Alkylhalogenide sind, aufgrund der Aktivierung, verursacht durch das Ethersauerstoffatom, 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 which show specificity against amino groups in the absence of reactive thiol groups and of the type XH ₂ CO- (where X = Cl, Br or I), e.g. B. as described by Wong, Y.-HH in Biochemistry (1979), 24, 5337;
• ii) N-Maleimide derivatives which are reacted with amino groups either by 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 nitrohaloaromatic 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 are usually stabilized by reduction to obtain a stable amine;
• vi) epoxy 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 very reactive towards nucleophiles such as amino, sulfhydryl and hydroxy groups;
• viii) aziridines based on the s-triazine compounds detailed above, e.g. B. as described by Ross, WCJ in Adv. Cancer Res. (1954) 2, 1, which react with nucleophiles such as amino groups by ring opening;
• ix) diethyl squarate, 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, due to the activation caused by the ether oxygen atom, e.g. B. as described by Benneche, T. et al., In Eur. J. Med. Chem. (1993) 28, 463.

• i) Isocyanate und Isothiocyanate, insbesondere aromatische Derivate, welche stabile Harnstoff- bzw. Thioharnstaff-Derivate bilden und für Protein-Vernetzung verwendet worden sind, wie beschrieben von Schick, A. F. et al. in Biol. Chem. (1961) 236, 2477;
• ii) Sulfonylchloride, welche beschrieben worden sind von Herzig, D. J., et al. in Biopolymers (1964) 2, 349, und welche für die Einführung einer fluoreszenten Reportergruppe in den Linker nützlich sein können;
• iii) Säurehalogenide;
• iv) aktive Ester, wie Nitrophenylester oder N-Hydroxysuccinimidylester;
• v) Säureanhydride, wie gemischte, symmetrische oder N-Carboxyanhydride;
• vi) andere nützliche Reagenzien für die Bildung von Amidbindungen, wie beschrieben von Bodansky, M., et al. in Principles of Peptide Synthesis' (1984) Springer-Verlag;
• vii) Acylazide, z. B. worin die Azidgruppe aus einem vorgeformten Hydrazid-Derivat unter Verwendung von Natriumnitrit erzeugt wird, z. B. wie beschrieben von Wetz, K., et al. in Anal. Biochem. (1974) 58, 347;
• viii) Azlactone, angeheftet an Polymere, wie Bisacrylamid, z. B. wie beschrieben von Rasmussen, J. K., in Reactive Polymers (1991) 16, 199; und
• ix) Imidoester, welche bei Reaktion mit Aminogruppen stabile Amidine bilden, z. B. wie beschrieben von Hunter, M. J., und Ludwig, M. L., in J. Am. Chem. Soc. (1962) 84, 3491.

Representative amine-reactive acylating agents include:

• i) isocyanates and isothiocyanates, especially aromatic derivatives, which form stable urea or thiourea derivatives and have been used for protein crosslinking, as described by Schick, AF et al. in Biol. Chem. (1961) 236, 2477;
• ii) sulfonyl chlorides which have been described by Herzig, DJ, et al. in Biopolymers (1964) 2, 349, and which may be useful for the introduction of a fluorescent reporter group into the linker;
• iii) acid halides;
• iv) active esters, such as nitrophenyl ester or N-hydroxysuccinimidyl ester;
• v) acid anhydrides such as mixed, symmetrical or N-carboxy anhydrides;
• vi) other useful reagents for the formation of amide bonds as described by Bodansky, M., et al. in Principles of Peptide Synthesis' (1984) Springer-Verlag;
• vii) acyl azides, e.g. B. wherein the azide group is generated from a preformed hydrazide derivative using sodium nitrite, e.g. B. as described by Wetz, K., et al. in anal. Biochem. (1974) 58, 347;
• viii) azlactones attached to polymers such as bisacrylamide, e.g. B. as described by Rasmussen, JK, in Reactive Polymers (1991) 16, 199; and
• ix) imido esters which form stable amidines when reacted with amino groups, e.g. B. as described by Hunter, MJ, and Ludwig, ML, in J. Am. Chem. Soc. (1962) 84, 3491.

Carbonyl groups, such as aldehyde functions, can be implemented with weak protein bases at a pH value, so that nucleophile Protein side chain functions are protonated. Weak bases include 1,2-aminothiols such as those found in N-terminal cysteine residues which selectively stable 5-membered thiazolidine rings with aldehyde groups form, e.g. B. as described by Ratner, S., et al. in J. Am. Chem. Soc. (1937) 59, 200. Other weak bases, such as phenylhydrazones, can are used, e.g. B. as described by Heitzman, H. et al., in proc. Natl. Acad. Sci. USA (1974) 71, 3537.

Aldehydes and ketones can too with amines to form Schiff bases, which can be advantageously stabilized by reductive amination. Alkoxylamino units react easily with ketones and aldehydes to produce more stable Alkoxamines, e.g. B. as described by Webb, R., et al. in Bioconjugate Chem. (1990) 1, 96.

Examples of reactive units, capable of Reaction with carboxyl groups, include diazo compounds such as Diazoacetatester and Diazoacetamide, one, which with high specificity under Generation of ester groups react, e.g. B. as described by Herriot, R.M., in Adv. Protein Chem. (1947) 3, 169. Carboxylic acid modifying reagents, like carbodiimides, which about O-acylurea formation, followed by amide bond formation, can also in useful Way to be used; the link can be created by adding a Amines can be facilitated or can be used for direct vector receptor coupling to lead. helpful water-soluble Close carbodiimides 1-Cyclohexyl-3- (2-morpholinyl-4-ethyl) carbodiimide (CMC) and 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide (EDC) a, e.g. B. as described by Zot, H.G., and Puett, D., in J. Biol. Chem. (1989) 264, 15552. Other useful carboxylic acid modifying reagents conclude Isoxazolium derivatives such as Woodwards reagent K; chloroformates, such as p-nitrophenyl chloroformate; Carbonyldimidazoles such as 1,1'-carbonyldiimidazole; and N-carbalkoxy dihydroquinolines, such as N- (ethoxycarbonyl) -2-ethoxy-1,2-dihydroquinoline, on.

Other potentially useful reactive moieties include vicinal diones, such as p-phenylene diglyoxal, which can be used to react with guanidinyl groups, e.g. B. as described by Wagner et al. in Nucleic Acid Res. (1978) 5, 4065; and diazonium salts, which may undergo electrophilic substitution reactions, e.g. B. as described by Ishizaka K., and Ishizaka, T., in J. Immunol. (1960) 85, 163, a. Bisdiazonium compounds are readily prepared by treating aryl diamines with sodium nitrite in acidic solutions. It is believed that functional groups in the reporter and / or vector can be converted to other functional groups as needed prior to the reaction, e.g. B. to impart additional reactivity or selectivity. Examples of methods useful for this purpose include converting amines to carboxylic acids using reagents such as dicarboxylic acid anhydrides; Conversion of amines to thiols using reagents such as N-acetylhomocysteine thiolactone, S-acetylmercapto-succinic anhydride, 2-iminothilane 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.

The vector reporter coupling can also using enzymes as cross-linking agents with zero length become; for example, transglutaminase, peroxidase and Xanthine oxidase has been used to make cross-linked products. Reverse proteolysis can also be used for crosslinking via amide formation become.

A non-covalent vector reporter coupling can, for example, by electrostatic charge interactions, via chelation in the form of stable metal complexes or via high affinity binding interaction be effected.

A vector attached to a peptide, Lipooligosaccharide or lipopeptide linker is coupled, which is an element contains that capable to mediate a membrane insertion can also be used become. An example is given by Leenhouts, J.M., et al. in Febs Letters (1995) 370 (3), 189-192, described.

Coupling can also be done using by avidin or streptavidin, which have four high affinity binding sites for biotin exhibit. Avidin can therefore be used to attach the vector to the To conjugate reporters when both vector and reporter biotinylate are. Examples are described by Bayer, E.A., and Wilchek, M., in Methods Biochem. Anal. (1980) 26, 1. This method can also be extended to the link from reporter to reporter, a process that may promote association of the agent and subsequent potentially increased effectiveness. Alternatively, you can Avidin or streptavidin attached directly to the surface of reporter particles become.

Non-covalent coupling can also use the bifunctional nature of bispecific immunoglobulins. These molecules can specifically bind two antigens, thereby linking them. For example, either bispecific IgG or chemically constructed bispecific F (ab) ' ₂ fragments can be used as linking agents. Heterobifunctional bispecific antibodies have also been reported to link two different antigens, e.g. B. 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 (e.g., as described by Chen, Aa et al. In Am. J. Pathol. (1988) 130 , 216), are crosslinked by antibody molecules and lead to the formation of crosslinked assemblies of agents of the formula I with potentially increased effectiveness.

So-called zero-length connecting means, which is a direct covalent linkage of two reactive chemical groups without the introduction of additional linking material induce (e.g. as in the amide bond formation, which under Use of carbodiimides or induced enzymatically will) as required according to the invention be used, as well as agents such as biotin / avidin systems, which induce a non-covalent reporter-vector linkage and agents that induce electrostatic interactions.

The most common is the linking agent however comprise two or more reactive units, e.g. B. how described above, which by a spacer element are connected. The presence of such a spacer or Spacers allows bifunctional Linker with specific functional groups within a molecule or react between two different molecules, resulting in one Binding between these two components results and being extrinsic Linker-derived material is introduced into the reporter-vector conjugate. The reactive units in a linking agent can be the same (homobifunctional Means) or different (heterobifunctional means or where several dissimilar reactive units are present, heteromultifunctional means) be creating a diversity of potential reagents is provided which is a covalent Binding between any chemical species, either intramolecular or intermolecular can.

The nature of the through the linking agent introduced, extrinsic material can have a critical impact on the Goal direction assets and overall stability of the ultimate product. So it can be desirable be to introduce unstable bonds, which z. B. contain spacer arms that are biodegradable or are chemically sensitive or the enzymatic cleavage sites include. Alternatively, the spacer can be polymeric components lock in, z. B. to act as surfactants and to increase the stability of the agent. The Spacer can also contain reactive units, e.g. B. as described above, about surface networking to reinforce.

Spacer elements can typically consist of aliphatic chains, which are the reactive ones Separate units of the linker effectively by distances of between 5 and 30 Å. They can also include macromolecular structures, such as poly (ethylene glycols). Such polymeric structures, referred to herein as PEGs, are simple, neutral polyethers which have received great attention in biotechnical and biomedical applications (see, for example, Milton Harris, J. (ed.) "Poly (ethylene glycol) chemistry, biotechnical and biomedical applications ", Plenum Press, New York 1992). PEGs are soluble in most solvents, including water, and are highly hydrated in aqueous environments, with two or three water molecules attached to each ethylene glycol segment; this has the effect of preventing the adsorption of either other polymers or proteins on PEG-modified surfaces. PEGs are known to be non-toxic and damage inactive proteins or cells, while covalently linked PEGs are known to be non-immunogenic and non-antigenic. In addition, PEGs can be easily modified and bound to other molecules with little impact on their chemistry. Their advantageous solubility and biological properties are evident from the many possible uses of PEGs and copolymers thereof, including block copolymers such as PEG polyurethanes and PEG polypropylenes.

Suitable molecular weights for according to the invention PEG spacers used can for example between 120 Dalton and 20 kDalton.

The main mechanism for recording of particles through the cells of the reticuloendothelial system (RES) is opsonization by plasma proteins in the blood; this mark foreign particles, which are then taken up by the RES. The biological properties of PEG spacer elements which according to the invention can be used serve to reduce the cycle time of the agent to a similar one to that Way to increase which for PEGylated liposomes are observed (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 areas of interest can also be achieved using antibodies which are bound to the ends of PEG spacers (See, e.g., Maruyama, K., et al. in Biochim. Biophys. 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, such as polygalacuturonic acid, glycosaminoglycans, heparinoids, Cellulose and marine polysaccharides such as alginates, chitosans and carrageenans; Storage type polysaccharides such as starch, glycogen, Dextran and aminodextrans; Polyamino acids and methyl and ethyl esters thereof, as in homo- and copolymers of lysine, glutamic acid and aspartic acid; and polypeptides, oligosaccharides and oligonucleotides, which enzyme cleavage sites can contain or not, a.

In general, spacer elements can contain cleavable groups such as vicinal glycol, azo, sulfone, ester, thioester or disulfide groups. Spacers containing biodegradable methylene diester or diamide groups of the formula - (Z) _m .YXC (R ¹ R ² ) .XY (Z) _n - [wherein X and Z are selected from -O-, -S- and -NR- (where 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 zero or 1; and R ¹ and R ^{2 are} each hydrogen, an organic group or a group -XY (Z) _m - or together form a divalent organic group] may also be useful; such as discussed in WO-A-92/17436, such groups are easily grown in the presence of esterases, e.g. B. in vivo, biodegraded, but are stable in the absence of such enzymes. Therefore, they can advantageously be linked to therapeutic agents to allow slow release thereof.

Poly [N] - (2-hydroxyethyl) methacrylamides] are potentially useful Spacer materials thanks to its small size interaction with cells and tissues (see e.g. Volfová, I., Ríhová, B. and V. R., and Vetvicka, P., in J. Bioact. Comp. Polymers (1992) 7, 175-190). Working on a similar Polymer consisting mainly from the closely related 2-hydroxypropyl derivative, showed that it was from the mononuclear Phagocyte system endocytized to a fairly small extent (see Goddard, P., Williamson, I., Bron, J., Hutchinson, L.E., Nicholls, J., and Petrak, K., in J. Bioct. Compatl. Polym. (1991) 6: 4-24).

• i) Copolymere von Methylmethacrylat mit Methacrylsäure; diese können erodierbar sein (siehe Lee, P. I., in Pharm. Res. (1993) 10, 980), und die Carboxylat-Substituenten können einen höheren Grad an Quellung als bei neutralen Polymeren verursachen;
• ii) Blockcopolymere von Polymethacrylaten mit biologisch abbaubaren Polyestern (siehe z. B. San Roman, J., und Guillen-Garcia, P., in Biomaterials (1991) 12, 236–241);
• iii) Cyanacrylate, d. h. Polymere von Estern von 2-Cyanacrylsäure – diese sind biologisch abbaubar und sind in der Form von Nanoteilchen für selektive Arzneimittelzuführung verwendet worden (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, welche wasserlöslich sind und im allgemeinen als biologisch kompatibel angesehen werden (siehe z. B. Langer, R., in J. Control. Release (1991) 16, 53–60);
• v) Copolymere von Vinylmethylether mit Maleinsäureanhydrid, welche angegebenermaßen biologisch erodierbar bzw. abbaubar sind (siehe Finne, U., Hannus, M., und Urtti, A., in Int. J. Pharm. (1992) 78, 237–241);
• vi) Polyvinylpyrrolidone, z. B. mit einem Molekulargewicht von weniger als etwa 25 000, welche rasch von den Nieren filtriert werden (siehe Hespe, W., Meier, A. M., und Blankwater, Y. M., in Arzneim.-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 Copolymeren, welche aromatische Hydroxysäuren beinhalten, um ihre Abbaurate zu erhöhen (siehe Imasaki, K., Yoshida, M., Fukuzaki, H., Asano, M., Kumakura, M., Mashimo, T., Yamanaka, H., und Nagai, T. in Int. J. Pharm. (1992) 81, 31–38);
• viii) Polyester, bestehend aus abwechselnden Einheiten von Ethylenglykol und Terephthalsäure, z. B. Dacron^®, welche nicht-abbaubar aber in hohem Maße biokompatibel sind;
• ix) Blockcopolymere, umfassend biologisch abbaubare Segmente von aliphatischen Hydroxysäure-Polymeren (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), beispielsweise in Verbindung mit Polyurethanen (siehe Kobayashi, H., Hyon, S. H., und Ikada, Y., in "Water-curable and biodegradable prepolymers" – J. Biomed. Mater. Res. (1991) 25, 1481–1494);
• x) Polyurethane, welche bekanntermaßen in Implantaten gut toleriert werden und welche mit flexiblen "weichen" Segmenten, z. B. umfassend Poly(tetramethylenglykol), Poly(propylenglylcol) oder Poly(ethylenglykol)), und aromatischen "harten" Segmenten, z. B. umfassend 4,4'-Methylenbis(phenylenisocyanat), kombiniert werden können (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) 80, 445–452, und Affrossman, S., et al. in Clinical Materials (1991) 8, 25–31);
• xi) Poly(1,4-dioxan-2-one), welche als biologisch abbaubare Ester in Hinsicht auf ihre hydrolysierbaren Esterbindungen angesehen werden können (siehe z. B. Song, C. X., Cui, X. M., und Schindler, A., in Med. Biol. Eng. Comput. (1993) 31, S147–150) und welche Glycolid-Einheiten einschließen können, um ihre Absorbierbarkeit zu verbessern (siehe Bezwada, R. S., Shalaby, S. W., und Newman, H. D. J., in Agricultural and synthetic polymers: Biodegradability and utilization (1990) (Hrsg.: Glass, J. E., und Swift, G.), 167–174 – ACS-Symposium-Serie, #433, Washington D.C., U.S.A, – American Chemical Society);
• xii) Polyanhydride, wie Copolymere von Sebacinsäure (Octandisäure) mit Bis(4-carboxyphenoxy)propan, von welchen in Kaninchenversuchen (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 in Rattenversuchen (siehe Tamargo, R. J., Epstein, J. I., Reinhard, C. S., Chasin, M., und Brem, H., in J. Biomed. Mater. Res. (1989) 23, 253–266) gezeigt worden ist, für die gesteuerte Freisetzung von Arzneimitteln im Gehirn ohne offensichtliche toxische Effekte nützlich zu sein;
• xiii) Biologisch abbaubare Polymere, enthaltend Ortho-Estergruppen, welche für die gesteuerte Freisetzung in vivo verwendet worden sind (siehe Maa, Y. F., und Heller, J., in J. Control. Release (1990) 14, 21–28); und
• xiv) Polyphosphazene, welche anorganische Polymere sind, bestehend aus abwechselnden Phosphor- und Stickstoffatomen (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 can be erodible (see Lee, PI, in Pharm. Res. (1993) 10, 980), and the carboxylate substituents can cause a higher degree of swelling than with neutral polymers;
• ii) block copolymers of polymethacrylates with biodegradable polyesters (see e.g. 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 delivery (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 to be biologically compatible (see, e.g., Langer, R., in J. Control. Release (1991) 16, 53-60);
• v) Copolymers of vinyl methyl ether with maleic anhydride, which are stated to be biodegradable or biodegradable (see Finne, U., Hannus, M., and Urtti, A., in Int. J. Pharm. (1992) 78, 237-241) ;
• vi) polyvinyl pyrrolidones, e.g. B. with 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 Arzneimittel.Forsch./Drug Res. (1977) 27, 1158-1162);
• vii) Polymers and copolymers of short-chain aliphatic hydroxy acids, such as glycolic, lactic, butteric, Valerian and Capronic acids (see e.g. Carli, F., in Chim. Ind. (Milan) (1993), 75, 494 -9), including copolymers containing aromatic hydroxy acids to increase their degradation rate (see Imasaki, K., Yoshida, M., Fukuzaki, H., Asano, M., Kumakura, M., Mashimo, T., Yamanaka , H., and Nagai, T. in Int. J. Pharm. (1992) 81, 31-38);
• viii) polyester consisting of alternating units of ethylene glycol and terephthalic acid, e.g. B. Dacron ^® , which are non-degradable but highly biocompatible;
• ix) block copolymers comprising biodegradable segments of aliphatic hydroxy acid polymers (see e.g. Younes, H., Nataf, PR, Cohn, D., Appelbaum, YJ, Pizov, G., and Uretzky, G., in Biomater Artif. Cells Artif. Organs (1988) 16, 705-719), for example in connection with polyurethanes (see Kobayashi, H., Hyon, SH, and Ikada, Y., in "Water-curable and biodegradable prepolymers" - J Biomed. Mater. Res. (1991) 25, 1481-1494);
• x) Polyurethanes, which are known to be well tolerated in implants and which have flexible "soft" segments, e.g. B. comprising poly (tetramethylene glycol), poly (propylene glycol) or poly (ethylene glycol)), and aromatic "hard" segments, e.g. B. comprising 4,4'-methylenebis (phenylene isocyanate) can be combined (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., et al. In J. Coll. Interface Sci. (1981) 80, 445-452, and Affrossman, S., et al. In Clinical Materials (1991) 8, 25-31);
• xi) poly (1,4-dioxan-2-one), which can be regarded as biodegradable esters with regard to their hydrolyzable ester linkages (see e.g. Song, CX, Cui, XM, and Schindler, A., in Med. Biol. Eng. Comput. (1993) 31, S147-150) and which glycolide units can include to improve their absorbability (see Bezwada, RS, Shalaby, SW, and Newman, HDJ, in Agricultural and synthetic polymers : Biodegradability 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-carboxyphenoxy) propane, of which in rabbit experiments (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 in rat experiments (see Tamargo, RJ, Epstein, JI, Reinhard, CS, Chasin, M., and Brem, H ., in J. Biomed. Mater. Res. (1989) 23, 253-266) has been shown to be useful for the controlled release of drugs in the brain with no apparent 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 following tables list linking agents which are useful in targetable agents according to the invention can.

(Continuation)

(Continuation)

In addition to the straight-chain and branched PEG-like linker (e.g. Polyethylene glycols and others with 2 to 100 repeating units of ethylene oxide) linkers in the VLR system can independently be a chemical bond or the rest of a linking group. The term "residue of a linking group" as used herein refers to a unit that remains, results, 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 minimally affects the ability of the vector to bind to its receptor. However, it is specifically considered that such vector reactive groups can also react with functional groups that are typically found on relevant protein molecules. Thus, in one aspect, the linkers useful in the practice of this invention derive from those groups that can react with any relevant molecule that includes a vector, as described above, that contains a reactive group, regardless of whether there is one relevant molecule is a protein or not, thereby forming a linking group.

Preferred linking groups are derived from vector-reactive groups, which, without being restricted thereto, are selected from:
a group which is directly linked to carboxy, aldehyde, amine (NHR), alcohols, sulfhydryl groups, activated methylenes and the like on the vector, for example active halogen-containing groups, including e.g. B. chloromethylphenyl groups and chloroacetyl [ClCH ₂ C (= O) -] groups, activated 2- (leaving group-substituted) 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 which can easily react with modified vector molecules containing a vector reactive group, ie vectors with a reactive group, modified to contain reactive groups, such as those mentioned in (1) above, e.g. B. 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 can contain 1 to about 20 carbon atoms. The aryl moieties of the linking groups can contain from about 6 to about 20 carbon atoms; and
a group which can be linked to the vector containing a reactive group or to the modified vector as noted above by using a crosslinking agent. The residues of certain useful crosslinking agents, such as homobifunctional and heterobifunctional gelatin hardeners, bisepoxides or bisisocyanates, can become part of a linking group during the crosslinking reaction. However, other useful crosslinking agents can facilitate crosslinking, such as self-consuming 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 crosslinking agents, 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. When an amide bond is formed, the crosslinking agent first reacts selectively with the carboxyl group and is then cleaved out during the reaction of the "activated" carboxyl group with an amine to form an amide bond therebetween, thus covalently bonding the two units. An advantage of this procedure is that the cross-linking of similar molecules, e.g. B. vector to vector, is avoided, whereas the reaction of, for example, homobifunctional crosslinking agents is non-selective and undesired crosslinked molecules are obtained.

Preferred useful linking groups are derived from various heterobifunctional cross-linking reagents such as those listed in the Pierce Chemical Company's Immunotechnology Catalog - Section: Protein Modification (1995 and 1996). Useful, non-limiting examples of such reagents include: Sulfo-SMCC Sulfosuccimindiyl-4- (N-maleimidomethyl) cyclohexane-1-carboxylate. Sulfo-SIAB Sulfosuccinimidyl (4-iodoacetyl) aminobenzoate. Sulfo-SMPB Sulfosuccinimidyl 4- (p-maleimidophenyl) buryrat. 2-IT 2-iminothiolane. SATA N-succinimidyl-S-acetylthioacetate.

In addition to the foregoing description, the linking groups, total or partly, also constructed or derived from complementary nucleotide sequences and nucleotide residues, both naturally occurring and modified, preferably non-self-associating oligonucleotide sequences. Particularly useful, non-limiting reagents for incorporating modified nucleotide units, which contain reactive functional groups, such as amine and sulfhydryl groups, into an oligonucleotide sequence are commercially available, for example, from Clontech Laboratories Inc. (Palo Alto California) and include Uni-Link AminoModifier (catalog # 5190), Biotin-ON phosphoramidite (catalog # 5191), N-MNT-C6-AminoModifier (catalog # 5202), AminoModifier II (catalog # 5203), DMT-C6-3'Amine-ON (catalog # 5222), C6-ThiolModifier (Catalog # 5211) and the like. In one aspect, the linking groups of this invention are from the reaction of a reactive functional group, such as an amine or sulfhydryl group, as available in the Clontech reagents above, one or more of which have been incorporated into an oligonucleotide sequence, for example with one or more the vector-reactive groups described above, such as a heterobifunctional group on the vector.

By attaching two complementary oligonucleotide sequences, one to the vector and the other to the reporter, includes the resulting double The oligonucleotide then hybridized 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, e.g. B. Poly (L-lactide / DL-lactide) poly (glycolide)
L-lactide / glycolide copolymers
Polycaprolactone and its copolymers
polyanhydrides
Poly (orthoester)
polyphosphazenes
Long chain straight or branched lipids (& phospholipids), sugars and carbohydrates
Oligonucleotides (see above)
as well as mixtures of the above.

Used according to the invention linking agent are generally a shortcut from vector to reporter or reporter to reporter with a certain Degree of specificity cause and can also used to have one or more therapeutically active agents to adhere.

The present invention thus sees a tool for the therapeutic drug delivery in combination with the Vector-mediated steering of the product to the desired location in front. By "therapeutic" or "drug" is meant a remedy which has a beneficial effect on a specific disease in a living human or non-human animal.

According to the present invention Therapeutic compounds used can be found inside a molecular Aggregate or particulate Encapsulated linkers or on the encapsulating walls of a vesicular linker attached or inserted into it. So the therapeutic Connection to part of the surface, for example via covalent or ionic bonds are or can be physically mixed into an encapsulating material especially if the drug is similar to the material polarity or solubility has so that it is prevented from exiting the product before it is intended to work in the body should. The release of the drug can only be done by the wetting contact with blood after administration or initiated as a result of other internal or external influences be, e.g. B. Dissolution processes, which be catalyzed by enzymes or use if the linker reporter is a gas-containing vesicle.

The therapeutic substance can covalently to the encapsulating membrane surface of a vesicular linker using a suitable linking agent, e.g. B. as herein described, linked. So you can, for example, initially produce a phospholipid derivative to which the medicinal product is biodegradable bond (s) or linker is bound, and then bring this derivative into the material used to manufacture it the vesicle membrane as described above. Alternatively, the agent may initially be without the therapeutic Substance are produced, which are then used on particulate (z. B. vesicular) Means can be coupled or coated thereon. For example a therapeutic substance into a suspension of microbubbles aqueous media added and shaken to bind the therapeutic substance to the microbubbles or pin.

The therapeutic substance can z. B. be a drug or pro-drug known for use in fighting of restenosis, hyptertension, stroke, congestive heart failure or another cardiovascular Therapy, or any other drug or pro-drug with ET-fighting Effects.

By targeting an agent according to the invention containing a pro-drug activating enzyme in pathological regions, one can map the targeting of the enzyme, which is possible makes it possible to visualize when the remedy is directed correctly and when the remedy has disappeared from non-target areas. In this way one can determine the optimal time for injecting prodrugs into individual patients.

Another alternative is a pro-drug, a pro-drug activating enzyme and a vector in the same particulate Linker reporter to bring in such a way that the pro-drug only after a certain external stimulus will be activated. Such a stimulus can e.g. B. a burst of vesicles by external ultrasound, light stimulation of a chromophoric reporter or magnetic heating of a superparamagnetic Be reporters after the one you want Target steering has been achieved.

So-called pro-drugs can also in agents according to the invention be used. So can Medicinal products are derivatized or modified to their physicochemical Change properties and adapt to the means of the invention; such derivatized drugs can are considered pro-medicines and are usually inactive until the Cleavage of the derivatizing group the active form of the drug restores.

Therapeutics can easily be attached to the Heart and vascular system in general, and to the liver, spleen and kidneys and other regions, like the lymphatic system, body cavities or the gastrointestinal system.

If the reporter chelated or chelated metal species (e.g. a paramagnetic metal ion or a metal radionuclide), the linker can, for example a chain attached to a metal chelating group is a polymer chain with a variety of metal chelating Groups, attached of the molecular backbone or built into the molecular backbone, a branched polymer with metal chelating groups at the branching ends (e.g. a dendrimeric polychelant), etc. What of that Linker is only required to have the vector and reporter units while of a reasonable duration. With reasonable duration is meant a sufficient duration for the contrast medium to be desired Has effects z. B. to contrast in vivo during to reinforce a diagnostic imaging process.

Thus, it may be desirable in certain circumstances be that the Linker is biodegradable after administration. By selecting one Suitable biodegradable linkers, it is possible to use the pattern of biological Distribution and biological elimination for the vector and / or reporter to modify. If vector and / or reporter is biologically active are or are able to have undesirable effects if keep them back After the image acquisition process is over, it may be desirable to plan a biodegradability of the linker or by design provide adequate biological elimination or metabolic decomposition of the vector and / or reporter units ensures. So a linker can have a biodegradable function contain, which decomposition products with modified Patterns of the biological distribution, which results from the release of the Reporters from the vector or from the fragmentation of a macromolecular Structure result. As an example of linkers that chelated Wear metal ion reporter, it is possible to involve the linker to provide a biodegradable function, which after the Decomposition releases an excretable chelate compound that the Contains reporter. Hence can biodegradable functions, if desired, within the linker structure are involved, preferably at locations which (a) branch locations (b) are at or near vector or reporter attachment points, or (c) are such that the biodegradation is physiological tolerable or rapidly excreted fragments.

Examples of suitable biological close degradable functions Ester, amide, double ester, phosphoester, ether, thioether, Guanidyl, acetal and ketal functions on.

As discussed above, the linker group, if desired have groups built into their molecular backbone, which influence the biological distribution of the contrast medium or which is a suitable spatial Conformation for ensure the contrast medium, z. B. to access water to chelated paramagnetic metal ion reporters to allow. As an example, the basic linker framework can be partial or essentially complete consist of one or more polyalkylene oxide chains.

Thus, the linker can be regarded as a composite of optionally biodegradable vector binding (V _b ) and reporter binding (R _b ) groups, linked via linker backbone (L _b ) groups, the linker backbone groups linking linker side chain groups ( L _sc ) can wear to modify the biological distribution, etc., and in turn may include biodegradable functions. The R _b and V _b binding groups can be attached to the linker backbone or can be present at linker backbone ends, for example with an R _b or V _b group at an L _b terminus, with R _b - or V _b groups which bind two L _b termini to one another, or wherein an L _b term carries two or more R _b or V _b groups. The L _b and L _sc groups are expediently oligomeric or polymeric structures (e.g. polyesters, polyamides, polyethers, polyamines, oligopeptides, polypeptides, oligo- and polysaccharides, oligonucleotides, etc.), preferably structures with at least partially hydrophilic or lipophilic in nature, e.g. B. hydrophi le, amphiphilic or lipophilic structures.

The linker can be a low, medium or high molecular weight e.g. B. up to 2MD. In general become linkers with higher Molecular weight preferred when used with a plurality of vectors or reporters should be loaded, or if it is necessary Vector and reporter spatially to separate, or if it is the linker himself who plays a role in modification of the biological distribution. In general linkers, however, have a molecular weight of 100 to 100,000 D, especially 120 D to 20 kD.

Linker to vector conjugation and linker to reporter can be through any suitable chemical Conjugation technique done, e.g. B. covalent bond (e.g. ester or amide formation), metal chelation or another metal coordinative or ionic bond, again as described above.

Examples of suitable linker systems conclude the enhancer polychelant structures from US-A-5 364 613 and WO 90/12050, polyamino acids (e.g. polylysine), functionalized PEG, polysaccharides, glycosaminoglycans, dendritic polymers, as described in WO 93/06868 and by Tomalia et al. in Angew. Chem. Int. Ed. Engl. 29: 138-175 (1990), PEG chelating agent 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 include the chelating moiety. Alternatively, the chelated metal can be carried on or in a particulate reporter. In any event, conventional metal chelating groups, as are well known in the radiopharmaceutical and MRI contrast media art, can be used, e.g. B. linear, cyclic and branched polyamino-polycarboxylic acids and phosphorus oxyacid equivalents and other sulfur and / or nitrogen ligands known in the art, e.g. B. DTPA, DTPA-BMA, EDTA, DO3A, TMT (see for example US-A-5 367 080), BAT and analogs (see e.g. Ohmono et al., J. Med. Chem. 35: 157- 162 (1992) and Kung et al., J. Nucl. Med. 25: 326-332 (1984)), Neurolite's N ₂ S ₂ chelating agent ECD; MAG (see Jurisson et al. Chem. Rev. 93: 1137-1156 (1993)), HIDA, DOXA (1-oxa-4,7,10-triazacyclododecanetriacetic acid), NOTA (1,4,7-triazacyclononetriacetic acid), TETA (1,4,8,11-tetraazacyclotetradecanetetraacetic acid), THT (4 '- (3-amino-4-methoxyphenyl) -6.6''- bis (N', N'-dicarboxymethyl-N-methylhydrazino) - 2.2 ': 6', 2 '' - terpyridine) etc. In this regard, the reader is referred to the patent literature by Sterling Winthrop, Nycomed (including Nycomed Imaging and Nycomed Salutar), Schering, Mallinckrodt, Bracco and Squibb, which chelating agents for the diagnostic metals, e.g. B. in MR, X-ray and radiodiagnostic means. See, for example, US-A-4-4 647 447, EP-A-71564, US-A-4 687 659, WO 89/00557, US-A-4 885 363 and EP A-232751.

reporter

The reporter units in the contrast media of the invention be any unit, capable either for direct or indirect detection in an in vivo diagnostic imaging procedures taking place, e.g. B. units, what detectable radiation (e.g. from radioactive decay, Fluorescence excitation, spin resonance excitation, etc.) or emit causes can be emit same, units which local electromagnetic Influence fields (e.g. paramagnetic, superparamagnetic, ferrimagnetic or ferromagnetic species), units which radiant energy absorb or scatter (e.g. chromophores, particles (including gas or more liquid Vesicles), heavy elements and connections thereof etc.), and units, which produce a detectable substance (e.g. gas microbubble generators), etc.

A very wide variety of materials that are detectable by diagnostic imaging modalities 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 a material that can produce an echogenic material will normally be chosen. For X-ray imaging, the reporter will generally be or contain a heavy atom (e.g., with an atomic weight of 38 or above), for MR imaging, the reporter will either be an isotope with a non-zero nuclear spin (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 (e.g. a colored or uncolored particle), a light absorber or a light emitter, for magnetometric imaging the reporter will be have detectable magnetic properties, for electrical impedance imaging, the reporter will affect electrical impedance, and for scintigraphy, SPECT, PET, etc., the reporter will be a radionuclide.

Examples of suitable reporters are widely known from the diagnostic imaging literature, e.g. B. magnetic iron oxide parts, gas-containing vesicles, chelated paramagnetic metals (such as Gd, Dy, Mn, Fe etc.); see for example US-A-4 647 447, WO 97/25073, US-A-4 863 715, US-A-4 770 183, WO 96/09840, WO 85/02772, WO 92/17212, WO 97/29783 , EP A-554213, US-A-5 228 446, WO 91/15243, WO 93/05818, WO 96/23524, WO 96/17628, US-A-5 387 080, 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 agent groups (for example tetraazacyclododecane chelating agents, such as DOTA, DO3A, HP-DO3A and analogues thereof) or by Linker chelating groups such as DTPA, DTPA-BMA, EDTA, DPDP, etc .; ^{Metal radionuclide} 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 300-1400 nm, especially 600 nm to 1200 nm, in particular 650 to 1000 nm; Vesicles containing fluorinated gases (ie containing materials in the gas phase at 37 ° C which contain fluorine, e.g. SF ₆ or perfluorinated C _1-6 hydrocarbons or other gases and gas precursors, enumerated in WO 97/29783); chelated heavy metal cluster ions (e.g. W or Mo polyoxo anions or the sulfur or mixed oxygen / sulfur analogs); covalently bonded non-metal atoms, which either have a high number of atoms (e.g. iodine) or are radioactive, e.g. B. ¹²³ I, ¹³¹ I atoms etc .; Vesicles containing an iodinated compound; Etc.

Generally speaking, the reporter (1) a chelatable metal or polyatomic, metal-containing ion (e.g. TcO, etc.) wherein the metal is a metal with a high atomic number (e.g. atomic number greater than 37), a paramagnetic species (e.g. a transition metal or lanthanide) or is a radioactive isotope, (2) a covalently bound non-metal species, which is an unpaired electron site (e.g. an oxygen or Carbon in a persistent free radical), a non-metal of high atomic number or a radioisotope, (3) a multi-atomic Cluster or crystal containing atoms of high atomic numbers, showing cooperative magnetic behavior (e.g. superparamagnetism, ferrimagnetism or ferromagnetism), or containing radionuclides, (4) a gas or a gas precursor (i.e. a material or a mixture of materials which) at 37 ° C gaseous is) (5) a chromophore (whereby with this term species which are fluorescent or phosphorescent, are included), z. B. an inorganic or organic structure, in particular a complexed metal ion or an organic group with an extended one delocalized electron system, or (6) a structure or group with the electrical impedance varying features, for. B. thanks of an extensive delocalized electron system.

Examples of particularly preferred ones Reporter groups are described in more detail below.

Chelated metal reporters: Metal radionuclides, paramanetic 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, ⁶⁷ Cu, ¹⁶⁷ Tm, ⁹⁷ Ru, ¹⁸⁸ Re, ¹⁷⁷ Lu, ¹⁹⁹ Au, ²⁰³ Pb and ¹⁴¹ Ce.

Preferred paramagnetic metal ions conclude Ions of transition and lanthanide metals (e.g. metals with atomic numbers from 6 to 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, especially from Mn, Cr, Fe, Gd and Dy, more specifically Gd.

Preferred fluorescent metal ions conclude Lanthanides, in particular La, Ce, Pr, Nd, Pm, Sm, Eu, Gd, Tb, Dy, Ho, Er, Tm, Yb and Lu a. Eu is especially preferred.

Preferred heavy metal containing Reporters can Include atoms of Mo, Bi, Si and W, and in particular can be multi-atom 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 unit or in or on a particle (e.g., 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 chelating groups are disclosed in US-A-4 647 447, WO 89/00557, US-A-5 367 080, US-A-5 364 613 etc.

The linker unit or the particle can contain one or more such chelating groups, if required metalized by more than one metal species (e.g. so that reporters are provided, which can be demonstrated in different imaging modalities are).

Especially if the metal is non-radioactive , it is preferred that a Polychelant linker or a particulate reporter can be used.

A chelating or chelating or complexing group, as specified herein, the rest one or more of a wide variety of chelating agents which comprise a metal ion or a polyatomic ion (e.g. TcO) can complex.

As is well known, is a chelating agent a compound containing donor atoms, which by coordinative Bond with a metal atom to form a cyclic structure, referred to as a chelating complex or chelate can. This class of compounds is published in the Kirk-Othmer Encyclopedia of Chemical Technology, Vol. 5, 339-368.

The remainder of a suitable chelating agent can be composed of polyphosphates such as sodium tripolyphosphate and hexametaphosphoric acid; Amino carboxylic 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 thenoyl trifluoroacetone; Hydroxy carboxylic 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 1,0-phenanthroline; Phenols such as salicylaldehyde, disulfopyrocatechol and chromotropic acid; Aminophenols such as 8-hydroxyquinoline and oxime sulfonic acid; Oximes, such as dimethylglyoxime and salicylaldoxime, peptides containing a proximal chelating functionality such as polycysteine, polyhistidine, polyaspartic acid, polyglutamic acid or combinations of such amino acids; Schiff bases such as disalicylaldehyde, 1,2-propylene diimine; Tetrapyrroles such as tetraphenylporphin and phthalocyanine; Sulfur compounds such as toluenedithiol, meso-2,3-dimercaptosuccinic acid, dimercaptopropanol, thioglycolic acid, potassium ethyl xanthate, 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 hydroxyethylidene diphosphonic acid, or combinations of two or more of the above agents can be selected. The remainder 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" - diethylene-triaminepentaacetic 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) -cyclohexanediethylene-triamine-pentaacetic acid (CDTPA).

Other suitable residues of chelating agents include proteins modified for the chelation of metals such as technetium and rhenium as described in U.S. Patent No. 5,078,985.

Suitable residues of chelating agents can also be derived from compounds containing N3S and N2S2, such as B. those disclosed in U.S. Patent Nos. 4,444,690; 4,670,545; 4,673,562; 4,897,255; 4,965,392; 4,980,147; 4,988,496; 5 021 556 and 5 075 099.

Other suitable residues of chelating agents are described in PCT / US 91/08253.

Preferred chelating group will be chosen from the group, consisting of 2-aminomethylpyridine, imino acetic acid, iminodiacetic acid, ethylenediaminetetraacetic (EDTA), diethylenetriaminepentaacetic acid (DTPA), 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA), Carbonyliminodiacetic acid, methyleniminoacetic acid, methyleniminodiacetic acid, ethylenethioethyleniminoacetic acid, ethylenethioethyleniminodiacetic acid, TMT, a terpyridinyl group, a chelating agent a terpyridyl group and a carboxymethylamino group, or a salt of any of the above acids. Especially preferred chelating groups are DTPA, DTPA-BMA, DPDP, TMT, DOTA and HPDO3A.

Representative chelating groups are also described in US 5,559,214 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.

Process for metallizing anyone Existing chelating agents are within the level of knowledge in the field. Metals can into any chelating unit by any of three general methods are introduced: direct introduction, matrix synthesis and / or Transmetallation. Direct contribution is preferred.

Therefore, it is desirable that the metal ion is easily complexed to the chelating agent, for example only by exposing or mixing an aqueous solution of the chelating agent-containing ones Unit with a metal salt in an aqueous solution, preferably with a pH in the range of about 4 to about 11. The salt can be any Salt, but preferably the salt is a water soluble salt of the metal, such as a halogen salt, and more preferred Salts chosen so that she do not interfere with the binding of the metal ion with the chelating agent. The The chelating agent-containing unit is preferably included in aqueous solution a pH of between about 5 and about 9, more preferably a pH between about 6 to about 8. The chelating agent-containing unit can mixed with buffer salts such as citrate, acetate, phosphate and borate to establish the optimal pH. Preferably be the buffer salts selected that she not the subsequent one Disrupt the binding of the metal ion to the chelating agent.

In diagnostic imaging contains the vector linker reporter (VLR) construct preferably a ratio from metal radionuclide ion to chelating agent contained in such Diagnostic imaging applications is effective. In preferred embodiments amounts the molar ratio of Metal ion per chelating agent to about 1: 1000 to about 1: 1.

In radiotherapeutic applications, the VLR preferably contains a metal-radi ratio onuclide ion to chelating agent, which 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 T1. 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 especially preferred. These radioisotopes can be atomic or, preferably, ionic.

The following isotopes or pairs of isotopes can be used for both imaging and therapy without the need to change the radiolabeling method or the chelating agent: 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

If the linker unit contains a single chelating agent, this chelating agent can be linked directly to the vector unit, e.g. B. via one of the metal coordinating groups of the chelating agent, which can form an ester, amide, thioester or thioamide bond with an amine, thiol or hydroxyl group on the vector. Alternatively, the vector and the chelating agent can be linked directly via a functionality which is bound to the chelating agent backbone, e.g. B. a CH ₂ phenyl NCS group attached to a ring carbon of DOTA as suggested by Meares et al. In JACS 110: 6266-6267 (1988), or indirectly via a homo- or heterobifunctional linker, e.g. B. a bis-amine, bis-epoxide, diol, diacid, difunctionalized PEG, etc. In this case, the bifunctional linker will expediently provide a chain of 1 to 200, preferably 3 to 30 atoms between the vector and the chelating agent residue.

worin Ch eine Chelatbildner-Einheit ist und Li eine Linkergrundgerüst-Komponente ist, d. h. der Linker weist vorzugsweise anhängende Chelatbildner, im Grundgerüst befindliche Chelatbildner oder terminale Chelatbildner oder eine Kombination hiervon auf. Die anhängenden und im Grundgerüst befindlichen polymeren Strukturen können verzweigt sein, sind weiter vorzugsweise aber linear, und die wiederkehrenden Einheiten (LiCh) oder andere wiederkehrende Einheiten in dem Polymer können im Grundgerüst befindliche oder anhängende Gruppen zur Modifikation der biologischen Verteilung aufweisen, z. B. Polyalkylengruppen, wie in WO 94/08629, WO 94/09056 und WO 96/20754.If the linker unit contains a plurality of chelating groups, the linker is or preferably contains sections of the formula

wherein Ch is a chelating agent unit and Li is a linker backbone component, ie the linker preferably has attached chelating agents, chelating agents in the backbone or terminal chelating agents or a combination thereof. The pendant and backbone polymeric structures may be branched, but are more preferably linear, and the repeating units (LiCh) or other repeating units in the polymer may have backbone or pendant groups in the backbone to modify the biodistribution, e.g. B. polyalkylene groups, as in WO 94/08629, WO 94/09056 and WO 96/20754.

The terminal chelating structures Li (Ch) _n , which can be dendritic polymers as in WO 93/06868, can have groups for modifying the biological distribution which are bound to terms which are not occupied by chelating agents and can have sites for promotion of biodegradation within the linker structure, as in WO 95/28966.

The chelating units within of the polychelant linker can be functionalized via the framework the chelating agent or by using one or more of the metal coordinating groups of the chelating agent or by Amide or ether bond formation between acid chelator and one Amine- or hydroxyl-bearing linker backbone are bound, e.g. B. how in polylysine-polyDTPA, polylysine-polyDOTA and in the so-called Magnifier polychelants from WO 90/12050. Such polychelant linkers can one or more vector groups either directly (e.g. using of amine, acid or hydroxyl groups in the polychelant linker) or via one bifunctional linker connection as above for monochelate linker discussed was conjugated.

If the chelated species of a particulate (or molecular aggregate e.g. vesicular) linker is carried, the chelate can be, for example, an unbound mono- or polychelate be (such as Gd DTPA-BMA or Gd HP-DO3A) which is within the particle is included, or it can be a mono- or polychelate, which is attached to the particle either by 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 e.g. B. WO 96/11023) is conjugated.

Non-metal atomic reporters

Preferred non-metal atom 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 reporters, preferably one Majority of them, e.g. B. 2 to 200, can be covalently attached to a linker backbone either directly using conventional ones chemical synthesis techniques or via a carrier group, e.g. B. a triiodophenyl group, be bound.

In one embodiment of this invention the use of radioisotopes of iodine is specifically considered drawn. For example, if the vector or linker consists of substituents which is built up chemically by iodine in a reaction to form a covalent bond can be substituted, such as. B. substituents with a hydroxyphenyl functionality, such substituents by radioisotope methods well known in the art be marked by iodine. The iodine species can be used in therapeutic and diagnostic imaging applications are used. however can, at the same time, contain a metal in a chelating agent the same vector linker also in either therapeutic or diagnostic imaging applications.

As with those discussed above Metal chelators, can such metal-atom reporters bound to the linker or in or on a particular linker, z. B. in a vesicle (see WO 95/26205 and GB 9624918.0) worn become.

Linker of the above related The type described with the metal reporters can be used for non-metal atom reporters be used, the non-metal atom reporter or groups which such Reporters wear the place of some or all of the chelating groups take in.

organic chromophore or fluorophore reporter

Preferred organic chromophores and close fluorophore reporters Groups with an extensive delocalized electron system a, e.g. B. cyanines, merocyanines, phthalocyanines, naphthalocyanines, triphenylmethines, Porphyrins, pyrilium dyes, Thiapyrilium dyes, squarylium dyes, croconium dyes, Azulenium dyes, indoanilines, benzophenoxazinium dyes, Benzothiaphenothiazinium dyes, anthraquinones, naphthoquinones, Indathrenes, phthaloylacridones, trisphenoquinones, azo dyes, intramolecular and intermolecular charge transfer dyes and Dye complexes, tropones, tetrazines, bis (dithiolene) complexes, Bis (benzenedithiolate) complexes, iodaniline dyes, bis (S, O-dithiolene) complexes, etc. Examples of suitable organic or metallized organic Chromophores can can be found "Topics in Applied Chemistry: Infrared absorbing dyes ", ed. M. Matsuoka, Plenum, NY 1990," Topics in Applied Chemistry: The Chemistry and Application of Dyes ", Waring 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., Heterocyclic 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) and WO 96/17628. Specific examples of chromophores that can be used include xylene cyanol, Fluorescein, Dansyl, NBD, Indocyanine Green, DODCI, DTDCI, DOTCI and DDTCI a.

Groups are particularly preferred, which have absorption maxima between 600 and 1000 nm in order to Interference with hemoglobin absorption to avoid (e.g. xylene cyanole).

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

Chalcogenpyrylomethin-Farbstoffe, z. B. die Verbindungen 12 auf Seite 31 von Matsuoka (siehe oben), d. h.

worin Y = Te, Se, O oder NR;
Monochalcogenopyrylomethin-Farbstoffe, z. B. die Verbindungen 13 auf der Seite 31 von Matsuoka (siehe oben), d. h.

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

worin X = O, S oder Se;
Thiapyrilium-Farbstoffe, z. B, die Verbindungen 15 auf der Seite 32, und die Verbindung I auf Seite 167 von Matsuoka (siehe oben), d. h.

worin n = 1 oder 2;
Squarylium-Farbstoffe, z. B. die Verbindung 10 und Tabelle IV auf Seite 30 von Matsuoka siehe oben, d. h.

worin X = CH=CH, Y = H und R = Et,
X = S, Y = H und R = Et, und
X = CMe₂, Y = H, und R = Me,
und Verbindung 6, Seite 26, von Matsuoka (siehe oben), d. h.

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

worin 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 (siehe oben), d. h.

worin X = CH=CH, Y = H, und R = Et;
Azulenium-Farbstoffe, z. B. die Verbindung 8 auf Seite 27 von Matsuoka (siehe oben), d. h.

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

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

worin 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 der Seite 201 von Matusoka (siehe oben), d. h.

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

Indanthren-Pigmente, z. B.

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

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

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

worin 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 (siehe oben), d. h.

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

aromatische Nicht-Benzenoid-Farbstoffe, z. B. Verbindung 8, ein Tropon, auf der Seite 92 von Matsuoka (siehe oben), d. h.

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

worin X = p-Phenylen oder
X = p-Terphenylen, sowie Verbindung 10 auf Seite 92 von Matsuoka (siehe oben), d. h.

worin X = p-Biphenyl;
kationische Salze von Tetrazin-Radikal-Farbstoffen, z. B. Verbindung 11 auf Seite 92 von Matsuoka (siehe oben)

worin X = p-Phenylen;
Donor-Akzeptor-Intermolekular-Ladungstransfer-Farbstoffe, z. B. CT-Komplexe der Verbindungen 13b und 14a bis 14c auf Seite 93 von Matsuoka (siehe oben), d. h.

worin 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 (siehe oben), d. h.

worin X = S oder Se, und Y = Tetrachlor, Tetrabrom, 2,3-Dicarbonsäure, 2,3-Dicarbonsäureanhydrid oder 2,3-Dicarbonsäure-N-phenylimid;
Naphthochinon-Farbstoffe, z. B. die Verbindungen 2, 3 und 4 auf Seite 37 von Matsuoka (siehe oben), d. h.

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

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

Metallphthalocyanine, wie Phthalocyanine, enthaltend Aluminium, Silicium, Nickel, Zink, Blei, Cadmium, Magnesium, Vanadium, Cobalt, Kupfer und Eisen, z. B. Verbindung 1 in Tabelle III auf Seite 52 von Matsuoka (siehe oben), z. B.

worin, zum Beispiel, M = Mg;
Metallnaphthalocyanine, wie Naphthalocyanine, enthaltend Aluminium, Zink, Cobalt, Magnesium, Cadmium, Silicium, Nickel, Vanadium, Blei, Kupfer und Eisen, siehe Verbindung 3 in Tabelle III auf Seite 52 von Matsuoka (siehe oben), z. B.

worin, zum Beispiel, M = Mg;
Bis(dithiolen)-Metallkomplexe, umfassend ein Metallion, wie Nickel, Cobalt, Kupfer und Eisen, koordiniert an vier Schwefelatome in einem bis(S,S'-zweizähnigen) Ligandenkomplex, z. B. siehe Tabelle I auf Seite 59 von Matsuoka (siehe oben)

worin
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-CH₃-O-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)₂-N-Phenyl, R₂ = Phenyl, M = Ni, und
R₁ = p-(CH₃)₂-N-Phenyl, R₂ = p-H,N-Phenyl, M = Ni;
Bis(benzoldithiolat)-Metall-Komplexe, umfassend ein Metallion, wie Nickel, Cobalt, Kupfer und Eisen, koordiniert an vier Schwefelatome in einem Ligandenkomplex, z. B. siehe Tabelle III auf Seite 62 von Matsuoka (siehe oben), d. h.

worin
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-zweizähnige Indoanilin-Farbstoffe, umfassend ein Metallion, wie Nickel, Cobalt, Kupfer und Eisen, koordiniert an zwei Stickstoff- und zwei Sauerstoffatome von zwei N,O-zweizähnigen Indoanilin-Liganden, z. B. Verbindung 6 in Tabelle N auf Seite 63 von Matsuoka (siehe oben), z. B.

worin 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)-Metallkomplexe, umfassend ein Metallion, wie Nickel, Cobalt, Kupfer und Eisen, koordiniert an zwei Schwefelatome und zwei Sauerstoffatome in einem bis(S,O-zweizähnigen) Ligandenkomplex, z. B. siehe U.S.-Patent 3 806 462, z. B.

a-Diimin-dithiolen-Komplexe, umfassend ein Metallion, wie Nickel, Cobalt, Kupfer und Eisen, koordiniert an zwei Schwefelatome und zwei Imino-Stickstoffatome in einem gemischten S,S- und N,N-zweizähnigen Diliganden-Komplex, z. B. siehe Tabelle II auf Seite 180, zweite von unten, von Matsuoka (siehe oben) (siehe auch japanische Patente: 62/39 682, 63/126 889 und 63/139 303), z. B.

und
Tris(a-Diimin)-Komplexe, umfassend ein Metallion, koordiniert an sechs Stickstoffatome in einem Triliganden-Komplex, z. B. siehe Tabelle II auf Seite 180 von Matsuoka (siehe oben), 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 dye, e.g. B. the compounds 4a to 4g, Table II on page 26 of Matsuoka (see above).

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

Chalcogen pyrylomethine dyes, e.g. B. the connections 12 on page 31 of Matsuoka (see above), ie

wherein Y = Te, Se, O or NR;
Monochalcogenopyrylomethine dyes, e.g. B. the connections 13 on page 31 of Matsuoka (see above), ie

where n = 1 or 2;
Pyrilium dyes, e.g. B. the connections 14 (X = O) on page 32 of Matsuoka (see above), ie

where X = O, S or Se;
Thiapyrilium dyes, e.g. B, compounds 15 on page 32, and compound I on page 167 from Matsuoka (see above), ie

where n = 1 or 2;
Squarylium dyes, e.g. For example, see compound 10 and Table IV on page 30 of Matsuoka above, ie

where X = CH = CH, Y = H and 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 (see above), ie

where X = CH = CH, Y = H and R = Et;
Croconium dyes, e.g. B. Compound 9 and Table IV on page 30 of Matsuoka (see above), 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 (see above), ie

where X = CH = CH, Y = H, and R = Et;
Azulenium dyes, e.g. B. Connection 8 on page 27 of Matsuoka (see above), ie

Merocyanine dyes, e.g. B. Compound 16, R = Me, on page 32 of Matsuoka (see above), ie

Indoaniline dyes, such as copper and nickel complexes of indoaniline dyes, e.g. B. Connection 6 on page 63 of Matsuoka (see above), 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, e.g. B. as shown on page 201 of Matusoka (see above), ie

where X = O or S;
1,4-diaminoanthraquinone (N-alkyl) -3'-thioxo-2,3-dicarboximide, e.g. B. Compound 20, on page 41 of Matusoka (see above)

Indanthrene pigments, e.g. B.

see connection 21 on page 41 of Matsuoka (see above);
2-arylamino-3,4-phthaloylacridone dyes, e.g. B. Connection 22 on page 41 of Matsuoka (see above)

Trisphenoquinone dyes, e.g. B. Connection 23 on page 41 of Matsuoka (see above)

Azo dyes, e.g. B. the monoazo dye, compound 2 on page 90 of Matsuoka (see above), 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) nBU, R ₃ = OMe, and R ₄ = NHAc;
Azo dyes, e.g. B. the polyazo dye, compound 5 on page 91 of Matsuoka (see above), ie

intramolecular charge transfer donor acceptor infrared dyes, e.g. B. Compounds 6 and 7 on page 91 of Matsuoka (see above), ie

aromatic non-benzenoid dyes, e.g. B. Compound 8, a tropon, on page 92 of Matsuoka (see above), ie

Tetrazine radical dyes, e.g. B. Compound 9 on page 92 of Matsuoka (see above), ie

where X = p-phenylene or
X = p-terphenylene, and compound 10 on page 92 of Matsuoka (see above), ie

where X = p-biphenyl;
cationic salts of tetrazine radical dyes, e.g. B. Connection 11 on page 92 of Matsuoka (see above)

where X = p-phenylene;
Donor-acceptor intermolecular charge transfer dyes, e.g. B. CT complexes of compounds 13b and 14a to 14c on page 93 of Matsuoka (see above), 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, e.g. B. Compounds 12 (X = S or Se) on page 38 of Matsuoka (see above), ie

wherein X = S or Se, and Y = tetrachlor, tetrabromo, 2,3-dicarboxylic acid, 2,3-dicarboxylic acid anhydride or 2,3-dicarboxylic acid-N-phenylimide;
Naphthoquinone dyes, e.g. B. Compounds 2, 3 and 4 on page 37 of Matsuoka (see above), ie

metallized azo dyes such as azo dyes containing nickel, cobalt, copper, iron and manganese;
Phthalocyanine dyes, e.g. B. Compound 1 in Table II on page 51 of Matsuoka (see above), e.g. B.

Naphthalocyanine dyes, e.g. B. Compound 3 in Table II on page 51 of Matsuoka (see above), e.g. B.

Metal phthalocyanines such as phthalocyanines containing aluminum, silicon, nickel, zinc, lead, cadmium, magnesium, vanadium, cobalt, copper and iron, e.g. B. Compound 1 in Table III on page 52 of Matsuoka (see above), e.g. B.

where, for 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 (see above), e.g. B.

where, for 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, e.g. B. see Table I on page 59 of Matsuoka (see above)

wherein
R ₁ = R ₂ = CF ₃ , M = Ni,
R ₁ = R ₂ = phenyl, M = Pd,
R ₁ = R ₂ = phenyl, M = Pt,
R ₁ = C4 to C10 alkyl, R ₂ = H, M = Ni,
R ₁ = C4 to C10 alkyl, R ₂ = H, M = Pd,
R ₁ = C4 to C10 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 = pH, 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, e.g. B. see Table III on page 62 of Matsuoka (see above), ie

wherein
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 from two N, O-bidentate indoaniline ligands, e.g. B. Compound 6 in Table N on page 63 of Matsuoka (see above), e.g. B.

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, e.g. See, e.g., U.S. Patent 3,806,462, e.g. B.

a-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, e.g. For example, see Table II on page 180, second from below, from Matsuoka (see above) (see also Japanese patents: 62/39 682, 63/126 889 and 63/139 303), e.g. B.

and
Tris (a-diimine) complexes comprising a metal ion coordinated to six nitrogen atoms in a triligand complex, e.g. B. see Table II on page 180 of Matsuoka (see above), last compound, (see also Japanese patents: 61/20 002 and 61/73 902), e.g. B.

Representative examples of visible Close dyes Fluorescein derivatives, Rhodamine derivatives, coumarins, azo dyes, metallizable dyes, Anthraquinone dyes, benzodifuranone dyes, polycyclic aromatic carbonyl dyes, indigoid dyes, polymethine dyes, Azacarbocyanine dyes, hemicyanine dyes, barbiturates, diazahemicyanine dyes, Styryl dyes, diarylcarbonium dyes, triarylcarbonium dyes, Phthalocyanine dyes, quinophthalone dyes, triphenodioxazine dyes, Formazan dyes, phenothiazine dyes such as methylene blue, azure A, Azur B and Azur C, oxazine dyes, thiazine dyes, naphtholactam dyes, Diazahemicyanin dyes, azopyridone dyes, azobenzene dyes, "Mordent" dyes, acidic Dyes, basic dyes, metallized and pre-metallized Dyes, xanthene dyes, direct dyes, leuco dyes, which for Manufacture of bathochromatic dyes from those of the precursor leuco dyes shifted shades can be oxidized, and other dyes, such as those listed by Waring, D.R., and Hallas, G., in "The Chemistry and Application of Dyes ", Topics in Applied Chemistry, Plenum Press, New York, N.Y., 1990, on.

Additional dyes can be listed in Haugland, R. P., "Handbook of Fluorescent Probes and Research Chemicals ", sixth edition, Molecular Probes, Inc., Eugene OR, 1996.

Such chromophores and fluorophores can covalently either directly to the vector or to or within one Linker structure to be bound. Again, linkers of the above described type in connection with the metal reporters for organic Chromophores or fluorophores are used, the chromophores / fluorophores take the place of some or all of the chelating agent groups.

As with those discussed above Metal chelating agents can Chromophores / fluorophores carried in or on particulate linker units be, e.g. B. in or on a vesicle, or covalently to inert matrix particles which also act as a light scattering reporter can.

Particulate reporter or link reporter

The particulate reporter and linker reporter are generally divided into two categories - those in which the particle comprises a matrix or shell that carries or contains the reporter, and those those in which the particle matrix itself is the reporter. Examples of the first category are: vesicles (e.g. micelles, liposomes, microballoons and microbubbles) containing a liquid, gas or solid phase which contains the contrast-effective reporter, e.g. B. an echogenic gas or a precursor thereof (see for example GB 9700699.3), a chelated paramagnetic metal or radionuclide, or a water-soluble iodinated X-ray contrast agent; porous particles loaded with the reporter z. B. loaded with paramagnetic metal molecular sieve particles; and solid particles e.g. B. from an inert biologically tolerable polymer to which the reporter is bound or coated, for. B. dye-loaded polymer particles.

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

Preferred particulate reporter or close reporter linker superparamagnetic particles (see US-A-4770183, WO 97/25073, WO 96/09840, etc.), echogenic vesicles (see WO 92/17212, WO 97/29783, etc.), vesicles containing death (see WO 95/26205 and GB 96249180) and Dye-loaded polymer particles (see WO 96/23524).

The particulate reporters can do one or several vectors in direct or indirect connection their surfaces exhibit. In general, it is preferred to have a plurality (e.g. 2 to 50) of vector units per particle. Especially becomes appropriate one, next to the one you want Targeting vector, including flow retardant vectors attach to the particles, d. H. Vectors that have an affinity for the capillary lumen or other organ surfaces possessed, which is sufficient to prevent the passage of the contrast medium through the capillaries or the target organ but slow down is not sufficient to immobilize the contrast medium. Such flow retarding vectors (for example, described in GB 9700699.3) can also serve the Anchoring contrast media once it's bound to its target location Has.

The path by which to attach Vector obtained on particles will depend on the nature of the particle surface. For inorganic Particles can link to the particle, for example, through the interaction between a metal-binding group (e.g. a phosphate, phosphonate or Oligo- or polyphosphate group) on the vector or on a linker, which is bound to the vector. For organic (e.g. polymer) Particles can be attached directly by a covalent vector Binding between groups on the particle surface and reactive groups in the vector, e.g. B. amide or ester bond, or by covalent The vector and particles are bound to a linker. left of the above in connection with chelated metal reporters discussed Type can used, although the linker is generally not used to couple particles together.

For non-solid particles, e.g. B. droplets (for example from water-insoluble iodinated liquids, as described in US-A-5318767, US-A-5451393, US-A-5352459 and US-A-5569448) and vesicles, the linker can conveniently contain hydrophobic "anchors" - Groups, for example saturated or unsaturated C _12-30 chains, which penetrate the particle surface and bind the vector to the particle. For example, the linker, for phospholipid vesicles, can serve to covalently bind the vector to a phospholipid that is compatible with the vesicle membrane. Examples of a linker which binds to vesicles and inorganic particles are described in GB 9622368.0 and WO 97/25073.

In addition to the vectors, others can Groups to the particle surface be bound, e.g. B. Stabilizers (to prevent aggregation) and modifiers for the biological distribution, like PEG. Such groups become Example in WO 97/25073, WO 96/09840, EP-A-284549 and US-A-4904479 described.

Preferably the V-L-R agents of the invention the non-peptide endothelin receptor targeting vectors (like Bosentan or BMS 182874) directly or indirectly to a reporter have coupled, e.g. B. with covalently bound iodine radioisotopes with Metal chelates, either directly or via an organic linker group be linked or to a particulate Reporter or linker reporter, e.g. B. superparamagnetic crystals (optionally coated, e.g. as in WO 97/25073), or a Vesicles, e.g. B. a (e) gas) or iodized contrast medium containing) micelle, liposome or micro hollow pearl.

The means of the invention can Patients for imaging in sufficient quantities to be administered the wished To provide contrast with the respective imaging technology.

If the reporter is a metal, doses of 0.001 to 5.0 mmol of chelated imaging metal ion per kilogram of the patient's body weight are generally effective to achieve adequate contrast enhancements. For most MRI applications, preferred dosages of the imaging metal ion will be in the range of 0.02 to 1.2 mmol / kg body weight, whereas for X-ray applications, dosages of 0.05 to 2.0 mmol / kg will generally be effective around one Achieve X-ray attenuation. Preferred dosages for most X-ray applications are 0.1 to 1.2 mmol of the lanthanide or heavy metal compound / kg body weight. Where the reporter is a radionuclide, doses of 0.01 to 100 mCi, preferably 0.1 to 50 mCi, are normally per 70 kg body weight should be sufficient. Where the reporter is a superparamagnetic particle, the dosage will normally be 0.5 to 30 mg Fe / kg body weight. Where the reporter is a gas or gas producer, e.g. B. in a micro hollow bead, the dosage will normally be 0.05 to 100 ul gas per 70 kg body weight.

The dosage of the compounds of the Invention for A therapeutic application is dependent on the finding to be treated depend, but is generally in the order of 1 pmol / kg to 1 mmol / kg body weight lie.

The compounds of the present Invention can with conventional pharmaceutical or veterinary excipients formulated such as emulsifiers, fatty acid esters, gelling agents, Stabilizers, antioxidants, osmolality adjusters, buffers, pH adjusting agents, etc., and can in a form suitable for parenteral or enteral administration Form are present, for example for injection or infusion or for direct administration into a body cavity with an external one Emptying, for example the gastrointestinal tract, the bladder or the uterus. So you can the compounds of the present invention in conventional pharmaceutical forms of administration, such as tablets, capsules, powders, Solutions, Suspensions, dispersions, syrups, suppositories etc. are available. However, solutions Suspensions and dispersions in physiologically acceptable carrier media, for example water for Injections, in general, are preferred.

The compounds according to the present Invention can therefore for administration using physiologically acceptable carrier or excipients can be formulated in a manner that is entirely within the level of knowledge in the field. For example, the Compounds, optionally with the addition of pharmaceutically acceptable ones Excipients, in an aqueous medium suspended or dissolved the resulting solution or suspension is then sterilized becomes.

For the imaging of some areas of the body is the most preferred Administration of contrast media the parenteral, z. B. intravenous administration. Parenterally administrable forms, e.g. B. intravenous solutions should be sterile and free of physiologically unacceptable agents and should be low osmolality exhibit irritation or other adverse effects after the To minimize administration, and therefore the contrast medium preferably isotonic or slightly hypertensive. suitable Close vehicle aqueous vehicles one, which is usually for the Administration of parenteral solutions used, such as sodium chloride injection, Ringer injection, dextrose injection, Dextrose and sodium chloride injection, lactylated Ringer injection and other solutions, as described in Remington's Pharmaceutical Sciences, 15th edition, Easton: Mack Publishing Co., pp. 1405-1412 and 1461-1487 (1975) and The National Formulary XIV, 14th Edition, Washington: American Pharmaceutical Association (1975). The solutions can contain preservatives, antimicrobial agents, buffers and antioxidants which conventionally for parenteral solutions used, contain excipients and other additives, which are compatible with the chelates and which are in production, Storage or use of products will not interfere.

The agents of formula I can both therapeutically effective in the treatment of disease states as well as in in vivo imaging. So can the vector on the reporter units for example therapeutic Efficacy be z. B. thanks to the radiotherapeutic effect a radionuclide reporter, the effectiveness of a chromophore (or Fluorophore) reporter in a photodynamic therapy or the Vector unit chemotherapeutic effect.

The use of the means of formula I in the manufacture of therapeutic compositions and in Method for the therapeutic or prophylactic treatment of the body of man or a non-human animal are thus considered representative for further Viewed aspects of the invention.

In one embodiment, the contrast medium comprises the invention expediently a gas-containing or gas-generating material, preferably in suspension in an aqueous sluggish material or conjugated to one or more vectors, at least of which one is a vector V as defined above, e.g. B. an endothelin antagonist, is. The gas can take the form of microbubbles, stabilized stabilized by a monolayer of a film-forming surfactant through a matrix material that is different from a surfactant. The vectors can for example coupled to such a surfactant or matrix be and can be bioactive or non-bioactive. The vectors can be different Targeting specificities and, in a preferred embodiment, are of such a nature that that they interact with their receptors, but not the gas ones Tie vesicles tightly.

The present invention will now further referring to the following non-limiting Illustrated examples. Unless otherwise stated are all percentages based on weight.

Example 1 - Endothelin Receptor Binding Contrast Agent for the MR imaging

Connection 1

Lysine (0.1 g, 0.7 mmol) is added to a solution of 27-0-3- [2- (3-carboxyacryloylamino) -5-hydroxyphenyl] acryloyloxymycerone (prepared according to EP 628569 , 0.5 g, 0.7 mmol) and DCC (N, N'-dicyclohexylcarbodiimide} in dry DMF (N, N-dimethylformamide) are added and the reaction mixture is stirred at ambient temperature and TLC or thin layer chromatography follows Dispersion is left overnight at + 4 ° C. The dispersion is filtered and the solvent is rotary evaporated before the substance is purified by chromatography.

Connection 2

Diethylenetriamine pentaacetic acid dianhydride (17.9 g, 50 mmol) is dissolved in dry DMF and compound 1 (0.45 g, 0.5 mmol), dissolved in dry DMF is added. The reaction mixture is at an elevated temperature under nitrogen atmosphere touched. Connects to the reaction a TLC. The solvent is rotary evaporated and the substance is purified by chromatography.

Gd (III) chelate of compound 2

Gadolinium oxide Gd ₂ O ₃ (0.1 g, 0.2 mmol) is added to a solution of compound 2 (0.5 g, 0.4 mmol) in water and the mixture is heated at 95 ° C. After filtration, the solution is evaporated and dried in vacuo at 50 ° C.

Example 2 - Endothelin Receptor Binding Contrast Agent for the nuclear medicine

^99m Tc chelate from compound 2

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

Example 3 - Endothelin Receptor Binding Contrast Agent for nuclear medicine

An aqueous solution of ¹³¹ I ₂ (2 equivalents) and sodium perchlorate (1 equivalent) becomes an aqueous solution of 2-benzo (1,3) -dioxol-5-yl-3-benzyl-4- (4-methoxyphenyl) - 4-oxobut-2-enoate (prepared according to Example 351 of WO 95/05376, 1 equivalent) was added. The solution is rotary evaporated and the substance is purified by chromatography.

Example 4 - Endothelin Receptor Binding Contrast Agent for ultrasound and stray light imaging

Connection 3

DSPE (distearoylphosphatidylethanolamine) (0.5 g, 0.7 mmol) is added to a solution of 27-O-3- [2- (3-carboxyacryloylamino) 5-hydroxyphenyl] acryloyloxymycerone (prepared according to EP 628569 , 0.5 g, 0.7 mmol) and DCC in dry DMF were added. The reaction mixture is stirred at ambient temperature and TLC follows. The dispersion is left overnight at + 4 ° C. The dispersion is filtered and the solvent is rotary evaporated before the substance is purified by chromatography.

Gaseous microparticles comprising phosphatidylserine and compound 3

Phosphatidylserine (5 mg) becomes 5% propylene glycol glycerol in water (1 ml) added. The dispersion is warmed to not more than 80 ° C for 5 minutes and then to ambient temperature cooled. The dispersion (0.8 ml) is transferred to a vessel (1 ml) and compound 1 (0.1-1.0 mg) is added. The vessel becomes a few Brought to a roller table for hours. The headspace of the vessel is rinsed with perfluorobutane. The jar turns 45 In a "cap" or vessel mixer for seconds shaken, after which the sample is placed on a roller table. After centrifugation the lower phase is exchanged with water. The washing process becomes 2-3 repeated times.

Example 5 - Gas-filled microbubbles of distearoylphophatidylserine comprising a lipopeptide containing a vector with affinity for endothelin receptors, for targeted Ultrasound imaging

a) Synthesis of 4 '- [3,4-dimethyl-5-isoxazolyl) sulfamoyl] -succinanilinic acid

Succinic anhydride (1.00 g, 10.0 mmol) and 4-dimethylaminopyridine (122 mg, 1.00 mmol) were added to a solution of sulfisoxazole (Aldrich, 267 mg, 1.00 mmol) in DMF (10 ml). The reaction mixture was stirred at 80 ° C for 2 hours and then concentrated. The residue was taken up in 5% aqueous sodium bicarbonate solution and extracted with ethyl acetate. The aqueous solution was acidified with dilute hydrochloric acid and the organic material was extracted into ethyl acetate. The organic phase was washed with dilute hydrochloric acid, water and brine, treated with activated carbon and dried (MgSO ₄ ). The solution was filtered and concentrated to give 280 mg (76%) of a white solid. The structure was confirmed by ¹ H (300 MHz) and ¹³ C (75 MHz) NMR spectroscopy. Further characterization was carried out using MALDI mass spectrometry (ACH matrix), whereby an M + Na peak at m / z 390 and an M + K peak at m / z 406 were obtained as expected.

b) Synthesis of a lipopeptide which is functionalized with sulfisoxazole

The structure shown above was synthesized on a manual nitrogen wash bottle or "bubbler" device, with Fmoc-protected "Rink-Amide" MBHA resin (Novabiochem) on a 0.125 mmol scale was started using amino acids from Novabiochem, palmitic acid from Fluka and the connection from a) above. The coupling was using standard TBTU / HOBt / DIEA protocols carried out. Simultaneous removal of the peptide from the resin and deprotection of the Side chain protecting groups were contained in TFA for 2 hours 5% EDT and 5% water. Raw material was precipitated from ether. The product was analyzed by analytical HPLC (gradient 70-100% B over 20 minutes, A = 0.1% TFA / water and B 0.1% TFA / acetonitrile, flow rate 1 ml / min, column: Vydac 218TP54, detection UV 214 nm, retention time 27 minutes). Another characterization was done using MALDI mass spectrometry carried out, wherein m + H was obtained at m / z 1359; expected: 1356.

c) production of gas-filled microbubbles, comprising the compound from b) above

A solution of 1.4% propylene glycol / 2.4% Glycerin (1.0 ml) was added to a mixture of DSPS (Avanti, 4.5 mg) and the product from b) above (0.5 mg) added in a vessel. The mixture was sonicated for 5 minutes and then at 80 ° C for 5 minutes heated (whereby the vessel during the warming shaken was) and cooled. The headspace was flushed with perfluorobutane gas and the vessel was Shaken in a "cap" mixer for 45 seconds, followed of extensive washing with deionized water. A MALDI mass stack trometry showed no detectable level of the compound b) above in the last washing solution.

The incorporation of isoxazole-containing Lipopeptide in the blisters was confirmed by MALDI-MS as follows. Approximately 50 ul microbubbles were transferred to a clean jar, which approx. 100 μl Contained 90% methanol. The mixture was sonicated for 30 seconds and analyzed by MALDI-MS (ACH matrix), showing an M + H peak at m / z 1359 was obtained, which corresponded to the lipopeptide b).

Example 6 ¹³¹ I-Labeled Sulfisoxazole (Compound 4):

Sulfisoxazole was labeled ¹³¹ I according to the procedure provided with Pierce's IODO-BEADS iodine. Marking yield: 91%.

Example 7 - Tc Chelate of N- (MAG-3} sulfisoxazole

The title compound was, as in Scheme 1 outlined below, as described in Nuclear Medicine Communications 16 (1995) 942-957.

Claims (21)

Composition of formula I. VLR (I) wherein V is a non-peptide organic group with binding affinity for an endothelin receptor site, L is a linker unit or a bond, and R is a unit that is detectable by in vivo imaging of a human or animal body. The composition of claim 1, wherein V is the residue of a compound of formula II ∅-Bd-Rg (II) wherein ∅ is an optionally substituted phenyl group, optionally part of a fused polycyclic structure with up to 4 fused rings; Bd is a 1 to 5 atom long, optionally unsaturated organic bridge, with backbone atoms selected from C, N, S and O, optionally equipped with side chain substituents and optionally forms part of a condensed ring, and containing one

Unit wherein X is C or SO, n is 0 or 1 and is YO, S or an amino nitrogen atom; and Rg is a five or six membered, saturated or unsaturated, optionally substituted carbocyclic or heterocyclic ring, optionally part of a fused polycyclic structure with up to 5 fused rings; in which the optional substituents are selected from halogen and oxygen atoms, carbon and sulfur oxy acid groups, hydroxyl, alkyl, aryl, aralkyl, acyl, alkoxy, alkylenedioxy, alkylthio, alkylamino, acyloxy, aralkylamino, aralkyloxy -, acylthio, amino and acylamino groups and combinations thereof, and in which the ring heteroatoms are selected from O, N and S, aryl groups are mono- or bicyclic, carbocyclic or heterocyclic rings with up to 10 ring atoms, and no more than 7 rings and there are no more than 6 condensed rings. The composition of claim 1, wherein V is the residue of a compound of formula III

wherein R ¹ and R ² , which may be the same or different, are hydroxy, sulfuroxyacid, acylamino or carboxyacylamino groups. The composition of claim 1, wherein V is the residue of a compound of formula IV

wherein each R ³ , which may be the same or different, is hydrogen or an acyl group; R ^{4 is} a carboxy or acyl group; and R ^{5 is} hydrogen, hydroxy or an acyloxy group. The composition of claim 1, wherein V is the residue of a compound of formula V.

(wherein R ^{6 is} hydrogen or alkyl; R ^{7 is} aryl, aralkyl, arylcarbonyloxy or aralkylcarbonyl and each alkyl moiety can have 1 to 6 carbon atoms; Z is an optionally oxa-substituted C _1-6 alkylene group, optionally having a CH ₂ unit is replaced by an amine nitrogen or an oxygen atom

Z ^{10 is} hydrogen and R ^{8 is} hydrogen or an optionally substituted C _1-6 alkyl group). The composition of claim 1, wherein V is the residue of a compound of formula VI

(wherein the isoxazoyl group is attached at the 5- or 3-positions; R ^{9 is} an optionally substituted, optionally fused benzene ring, a fused pyridine ring, a phenyl group, or a carboxyl or amino group, the ring to which it is attached unsaturated or is a fused 2,2-propylidene group or an oxygen atom, the ring to which it is attached is saturated; and R ^{10 is} a straight or branched C _1-6 alkyl group or a halogen atom). The composition of claim 1, wherein V is the residue of a compound of formula VII

(wherein R ^{11 is} hydrogen or C _1-6 alkyl optionally substituted by oxyacid or other reactive substituents for vector linkage; R ^{12 is} C _1-6 alkyl optionally substituted by oxyacid or other reactive substituents for vector linkage; R ^{13 is} hydrogen, halogen or C _1-6 alkyl and R ^{14 is} hydrogen, C _1-6 alkoxy or methylenedioxy. The composition of claim 1, wherein V is the residue of a compound of formula VIII

(wherein Z '' is selected from alkylsulfonylaminocarbonyl, arylsulfonylaminocarbonyl, aryl, arylaminocarbonyl and carboxy; R ^{15 is} hydrogen, alkyl, oxyacid or another reactive group for vector linking; R ^{16 is} hydrogen, halogen, carboxyl, C _1-6 alkoxy- Is carbonyl or C _1-6 alkyl, alkoxy, hydroxyalkyl or haloalkyl; and R ^{17 is} hydrogen, halogen, nitro, carboxyl, aminocarbonyl, acetylamino, alkoxycarbonylamino, alkoxy or alkylenedioxy). The composition of claim 1, wherein V is the residue of a compound of formula IX or X.

(wherein each R ¹⁸ , which may be different or the same, is an alkoxy or alkylenedioxy group). The composition of claim 1, wherein V is the residue of a compound of formula XI

(wherein each R ¹⁸ , which may be the same or different, is an alkoxy or alkylenedioxy group). The composition of claim 1, V is the residue of a compound of Formula XII or Formula XIII

(wherein R ^{19 is} C _1-6 alkyl; R ^{18 is} as defined in claim 10; Z '''is O or CH ₂ ; and R ^{20 is} alkoxyaryloxy, hydroxyalkoxy or alkoxyaryl). The composition of claim 1, wherein V is the residue of a compound of formula XIV

(wherein R ^{19 is} as defined in claim 11; R ^{21 is} a 5- or 6-membered aryl group; and each R ²² , which may be the same or different, is hydrogen or a C _1-6 alkyl group). The composition of claim 1, wherein V is the residue of a compound of formula XV

(wherein R ^{18 is} as defined in claim 10; R ^{23 is} an aralkyl group; and R ^{24 is} an aralkyl group). The composition of claim 1, wherein V is the residue of a compound of formula XVI

(wherein R ^{25 is} an oxyacid or an ester; R ^{18 is} as defined in claim 10; and R ^{26 is} a hydrogen atom or an optionally substituted C _1-6 alkyl group). The composition of claim 1 selected from: i) a Gd (III) chelate complex of a DTPA conjugate of a lysine conjugate of 27-0-3- [2- (3-carboxyacryloylamino) -5-hydroxyphenyl] acryloyloxymycerone; ii) a Tc chelate complex of a DTPA conjugate of a lysine conjugate of 27-O-3- [2- (3-carboxyacryloylamino) -5-hydroxyphenyl] acryloyloxymycerone; iii) I ¹³¹ -labeled 2-benzo (1,3) dioxol-5-yl-3-benzyl-4- (4-methoxyphenyl) -4-oxobut-2-enoate; iv) a gas-containing vesicle bearing 27-O-3- [2- (3-carboxyacryloyl-amino) -5-hydroxyphenyl] acryloyloxymycerone; v) a gas-containing vesicle carrying a lipopeptide functionalized with sulfisoxazole; vi) I ¹³¹ labeled sulfisoxazole; and vii) a Tc chelate complex of N- (MAG-3) sulfisoxazole. A pharmaceutical composition comprising a Composition of formula I according to any one of claims 1 to 15, together with at least one pharmaceutically active carrier or Excipient. Use of a composition of formula I according to any of the claims 1 to 15 for the production of a contrast medium for use in a diagnostic procedure which involves the administration of the contrast medium to a living subject and creating an image of at least one Part of the subject included. Process for creating an image of a living human or non-human animal subject, the previous one a contrast agent was administered and an image was formed at least part of the subject in which the contrast agent was distributed, characterized in that as the contrast agent a composition of formula I according to any one of claims 1 to 15 is used. The method of claim 18, wherein the image is a nuclear imaging image. The method of claim 18, wherein the image is an ultrasound image is. Method of making a composition of formula I according to any one of claims 1 to 15, which method conjugating (i) an organic compound with binding affinity for one Endothelin receptor to (ii) a compound used in a diagnostic Imaging method is detectable, or a chelate compound and, if necessary, metalating chelate groups in the resulting conjugate with a metal ion used in a diagnostic imaging process is detectable.