ITMI20011167A1 - CYTOTOXIC OR RADIOACTIVE CONJUGATES CAPABLE OF BINDING TO OXYTOCIN RECEPTORS - Google Patents

CYTOTOXIC OR RADIOACTIVE CONJUGATES CAPABLE OF BINDING TO OXYTOCIN RECEPTORS Download PDF

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ITMI20011167A1
ITMI20011167A1 IT2001MI001167A ITMI20011167A ITMI20011167A1 IT MI20011167 A1 ITMI20011167 A1 IT MI20011167A1 IT 2001MI001167 A IT2001MI001167 A IT 2001MI001167A IT MI20011167 A ITMI20011167 A IT MI20011167A IT MI20011167 A1 ITMI20011167 A1 IT MI20011167A1
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conjugates
cytotoxic
radioactive
oxytocin
binding
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Gianni Bussolati
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Gianni Bussolati
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Priority to PCT/EP2002/005687 priority patent/WO2002098447A1/en
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
    • A61K47/64—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
    • A61K38/08—Peptides having 5 to 11 amino acids
    • A61K38/095—Oxytocins; Vasopressins; Related peptides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
    • A61K51/02—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by the carrier, i.e. characterised by the agent or material covalently linked or complexing the radioactive nucleus
    • A61K51/04—Organic compounds
    • A61K51/08—Peptides, e.g. proteins, carriers being peptides, polyamino acids, proteins
    • A61K51/084—Peptides, e.g. proteins, carriers being peptides, polyamino acids, proteins the peptide being oxytocin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
    • A61K51/02—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by the carrier, i.e. characterised by the agent or material covalently linked or complexing the radioactive nucleus
    • A61K51/04—Organic compounds
    • A61K51/08—Peptides, e.g. proteins, carriers being peptides, polyamino acids, proteins
    • A61K51/088—Peptides, e.g. proteins, carriers being peptides, polyamino acids, proteins conjugates with carriers being peptides, polyamino acids or proteins

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Description

Descrizione dell'invenzione industriale avente per titolo: Description of the industrial invention entitled:

"CONIUGATI CITOTOSSICI O RADIOATTIVI CAPACI DI LEGARSI AI RECETTORI DELL’OSSITOCINA” "CYTOTOXIC OR RADIOACTIVE CONJUGATES ABLE TO BIND TO OXITOCIN RECEPTORS"

La presente invenzione riguarda coniugati di analoghi dell' ossitocina (OT) con agenti citotossici o radioattivi, per la radioterapia e l“imaging” di tumori che sovraesprimono recettori dell' ossitocina (OTR). The present invention relates to conjugates of oxytocin analogues (OT) with cytotoxic or radioactive agents, for radiotherapy and "imaging" of tumors overexpressing oxytocin receptors (OTR).

L’invenzione riguarda in particolare coniugati di analoghi di ossitocina con chelanti di metalli radioattivi o paramagnetici. The invention relates in particular to conjugates of oxytocin analogues with radioactive or paramagnetic metal chelators.

I recettori dell' ossitocina sono espressi in diversi tipi di cellule in organi quali mammella, utero, cervello e sistema vascolare. Gli OTR sono inoltre espressi in tumori quali carcinomi della mammella, neuroblastomi, glioblastomi, carcinoma endometriale, osteosarcomi, coriocarcinomi, sarcoma di Kaposi, carcinomi della prostata e del polmone. Alcuni analoghi dell' ossitocina sono risultati attivi nell' inibire la crescita di neuroblastomi, glioblastomi, carcinomi della mammella e dell’ endometrio in vitro e in vivo (Int.J.Cancer, 66:817-820, 1996; ibidem, 72:340-344, 1997). Un ligando radioattivo o citotossico specifico per OTR potrebbe offrire nuove strategie diagnostiche e terapeutiche per tumori OTR-positivi. Un’analoga strategia è stata provata con successo nel caso di tumori che esprimono recettori per la somatostatina, coniugando il chelante DOTA a un gruppo amminico libero della somatostatina o di suoi analoghi (Cancer Res. 153:1-13, 2000; Clin. CancerRes, 5:1025-1033, 1999). Oxytocin receptors are expressed in different types of cells in organs such as the breast, uterus, brain and vascular system. OTRs are also expressed in tumors such as breast carcinomas, neuroblastomas, glioblastomas, endometrial carcinoma, osteosarcomas, choriocarcinomas, Kaposi's sarcoma, prostate and lung cancers. Some oxytocin analogues have been found to be active in inhibiting the growth of neuroblastomas, glioblastomas, breast and endometrial carcinomas in vitro and in vivo (Int. J. Chaner, 66: 817-820, 1996; ibidem, 72: 340 -344, 1997). A radioactive or cytotoxic ligand specific for OTR could offer novel diagnostic and therapeutic strategies for OTR-positive tumors. A similar strategy has been successfully tried in the case of tumors expressing somatostatin receptors, by conjugating the chelator DOTA to a free amino group of somatostatin or its analogues (Cancer Res. 153: 1-13, 2000; Clin. CancerRes , 5: 1025-1033, 1999).

Si è ora trovato che analoghi dell' ossitocina, in particolare analoghi aventi in posizione 8 un amminoacido con gruppi funzionalizzabili, possono essere coniugati a opportuni chelanti di metalli senza perdere l’affinità nei confronti di OTR. L’invenzione fornisce pertanto coniugati di analoghi dell’ossitocina aventi un residuo di amminoacido fimzionalizzabile in posizione 8 con agenti citotossici o radioattivi, per la radioterapia e l’”imaging” di tumori che sovraesprimono recettori dell’ossitocina (OTR). È preferito in particolare, come analogo di ossitocina, il composto noto con la sigla di LVT (lisina- vasotocina), descritto in Helv. Chim. Acta, 43:182-185, 1960, avente una lisina in posizione 8. Come agenti citotossici coniugabili a LVT si possono impiegare anticorpi, tossine, farmaci citotossici, chelanti di metalli radioattivi o paramagnetici, biotina, etc. Sono preferiti i chelanti di metalli quali DOTA, BOPTA, DOTMA, coniugabili mediante legami ammidici con LVT. It has now been found that analogues of oxytocin, in particular analogues having an amino acid with functionalizable groups in position 8, can be conjugated to suitable metal chelators without losing the affinity towards OTR. The invention therefore provides conjugates of oxytocin analogues having a functionalizable amino acid residue in position 8 with cytotoxic or radioactive agents, for radiotherapy and "imaging" of tumors that overexpress oxytocin receptors (OTR). In particular, the compound known by the abbreviation LVT (lysine-vasotocin), described in Helv, is preferred as oxytocin analogue. Chim. Acta, 43: 182-185, 1960, having a lysine in position 8. Antibodies, toxins, cytotoxic drugs, radioactive or paramagnetic metal chelators, biotin, etc. can be used as cytotoxic agents that can be conjugated to LVT. Metal chelators such as DOTA, BOPTA, DOTMA, which can be conjugated by amide bonds with LVT, are preferred.

I coniugati corrispondenti possono chelare metalli o alogeni radioattivi quali The corresponding conjugates can chelate metals or radioactive halogens such as

I chelati radioattivi potranno essere usati come traccianti radioterapici per la localizzazione e il trattamento di tumori che esprimono OTR, mentre i chelati paramagnetici saranno utili per l’imaging di risonanza magnetica degli stessi tumori o di organi e tessuti che esprimono OTR. Il coniugato dell’ invenzione può eventualmente essere marcato con un tracciante quale fluoro per la localizzazione dei tumori con tecniche di PET ( Positron Emission Tomography). Radioactive chelates can be used as radiotherapy tracers for the localization and treatment of tumors that express OTR, while paramagnetic chelates will be useful for magnetic resonance imaging of the same tumors or organs and tissues that express OTR. The conjugate of the invention can possibly be labeled with a tracer such as fluorine for the localization of tumors with PET (Positron Emission Tomography) techniques.

L’invenzione riguarda pertanto anche composizioni farmaceutiche e diagnostiche contenenti quantità efficaci dei coniugati di LVT o di altri analoghi analoghi dell’ ossitocina. The invention therefore also relates to pharmaceutical and diagnostic compositions containing effective amounts of LVT conjugates or other analogues of oxytocin.

Tali composizioni saranno somministrate per via parenterale, a dosi che dipenderanno ovviamente dal tipo di chelante e di metallo, dalla terapia o diagnosi da effettuare e dalle condizioni del paziente. These compositions will be administered parenterally, at doses which will obviously depend on the type of chelant and metal, on the therapy or diagnosis to be carried out and on the patient's condition.

I metodi di preparazione dei coniugati dell’invenzione sono convenzionali, simili ad esempio a quelli descritti per analoghi di somatostatina. The methods of preparation of the conjugates of the invention are conventional, similar for example to those described for somatostatin analogs.

I seguenti Esempi illustrano l’invenzione in maggior dettaglio. The following examples illustrate the invention in greater detail.

ESEMPIO 1 EXAMPLE 1

DOTA fu sciolto in DMSO a 80°C e la soluzione fu raffreddata in atmosfera di argon. Una soluzione di N-idrossi-2,5-pirrolidindione (NHS) in DMSO fu aggiunta goccia a goccia alla soluzione di DOTA sotto agitazione, seguita dall’aggiunta di N,N’-dicicloesilcarbodiimmide (DCC) in DMSO. Il rapporto molare DOTA:NHS:DCC era 1:1.4:0.8. DOTA was dissolved in DMSO at 80 ° C and the solution was cooled in an argon atmosphere. A solution of N-hydroxy-2,5-pyrrolidinedione (NHS) in DMSO was added drop by drop to the DOTA solution under stirring, followed by the addition of N, N-dicyclohexylcarbodiimide (DCC) in DMSO. The DOTA: NHS: DCC molar ratio was 1: 1.4: 0.8.

La miscela fu fatta reagire per una notte e poi filtrata per separare la dicicloesilurea formatasi. La coniugazione fra DOTA e LVT fu effettuata ad un rapporto molare di 50:1 aggiungendo un adeguato volume della soluzione di estere attivato di DOTA a una soluzione di LVT in tampone fosfato =0.1 M, pH 8. Dopo reazione per una notte, il coniugato fu purificato per mezzo di una colonna in fase inversa in un sistema FPLC accoppiato ad un rivelatore UV e a un radiorivelatore. Si applicò un metodo di gradiente lineare impiegando una soluzione di acqua distillata contenente lo 0,1% di TFA (solvente A) e metanolo (solvente B). Gli eluenti furono caricati ad un flusso di 4 ml/min. partendo dallo 0% di solvente A fino al 100% di solvente B in 37 mi. Due picchi corrispondenti ai coniugati di LVT con DOTA erano evidenti nel profilo UV. Il volume di ritenzione era di 8,4 mi per il primo composto (A) e 10,0 mi per il secondo (B), mentre LVT non coniugato eluiva a 7,0 mi nelle stesse condizioni. Un collettore di frazioni integrato effettuò il recupero di ogni composto. Ogni picco fu successivamente analizzato per spettrometria di massa MALDI-TOF. The mixture was reacted overnight and then filtered to separate the dicyclohexylurea formed. The conjugation between DOTA and LVT was carried out at a molar ratio of 50: 1 by adding an adequate volume of the activated ester solution of DOTA to a solution of LVT in phosphate buffer = 0.1 M, pH 8. After reaction overnight, the conjugate it was purified by means of a reversed phase column in an FPLC system coupled to a UV detector and a radio detector. A linear gradient method was applied using a distilled water solution containing 0.1% TFA (solvent A) and methanol (solvent B). The eluents were charged at a flow of 4 ml / min. starting from 0% of solvent A up to 100% of solvent B in 37 ml. Two peaks corresponding to the conjugates of LVT with DOTA were evident in the UV profile. The retention volume was 8.4ml for the first compound (A) and 10.0ml for the second (B), while the unconjugated LVT eluted at 7.0ml under the same conditions. An integrated fraction collector carried out the recovery of each compound. Each peak was subsequently analyzed by MALDI-TOF mass spectrometry.

ESEMPIO 2 - Coniugazione con EXAMPLE 2 - Conjugation with

diluiti in tampone acetato (0,1 M, pH 5.5) furono aggiunti a 0,07 μmol della frazione ottenuta dal picco A. La soluzione fu riscaldata per 25 minuti a 80°C e si valutò la resa di marcatura per FPLC. diluted in acetate buffer (0.1 M, pH 5.5) they were added to 0.07 μmol of the fraction obtained from peak A. The solution was heated for 25 minutes at 80 ° C and the labeling yield was evaluated by FPLC.

ESEMPIO 3 - Studi di affinità. EXAMPLE 3 - Affinity studies.

Per determinare le costanti di affinità di LVT e di DOTA-LVT furono effettuati esperimenti di competizione eterologa su membrane preparate da cellule renali di scimmia COS7 transientemente trasferiate con cDNA umano di OTR. To determine the affinity constants of LVT and DOTA-LVT, heterologous competition experiments were performed on membranes prepared from COS7 monkey kidney cells transiently transferred with human OTR cDNA.

Brevemente, le cellule elettroporate furono omogeneizzate, lavate due volte e risospese in tampone di legame (50mM Tris H delle proteine di membrana furono incubate con una concentrazione fissa di (1-2 nM) per 30 minuti a 30°C in presenza di concentrazioni crescenti di peptidi non marcati. I legame non specifico fu determinato in presenza di 1 mM OT. La radioattività legata e libera fu separata per filtrazione e le isoterme di legame furono analizzate con il programma iterativo di adattamento della curva Ligand. I dati riportati in figura 1 mostrano come LVT non marcato inibiva il legame a con alta affinità Briefly, the electroporated cells were homogenized, washed twice and resuspended in binding buffer (50mM Tris H of the membrane proteins were incubated with a fixed concentration of (1-2 nM) for 30 minutes at 30 ° C in the presence of increasing concentrations of unlabeled peptides. The non-specific binding was determined in the presence of 1 mM OT. The bound and free radioactivity was separated by filtration and the binding isotherms were analyzed with the iterative program of adaptation of the Ligand curve. The data reported in figure 1 show how unlabeled LVT inhibited high affinity a-binding

DOTA-LVT dal picco A era anche in grado di competere con il legame a con una Ki calcolata di 238 52.36 nM; n = 3. Le curve di spiazzamento erano parallele e le loro inclinazioni non erano significativamente diverse dall’unità. DOTA-LVT from peak A was also able to compete for a-binding with a calculated Ki of 238 52.36 nM; n = 3. The displacement curves were parallel and their inclinations were not significantly different from unity.

ESEMPIO 4 - Studi di legame su cellule tumorali OTR+ e OTR-. EXAMPLE 4 - Binding studies on OTR + and OTR- tumor cells.

Esperimenti di legame furono effettuati su cellule di carcinoma mammario (MCF7), di glioblastoma (MOG-U-V-W) e carcinoma del colon (HT29). 20 x 10<5 >cellule, sospese in 100 μΐ di terreno di cultura furono incubate per 30 minuti a 4°C in presenza di Binding experiments were performed on breast cancer (MCF7), glioblastoma (MOG-U-V-W) and colon cancer (HT29) cells. 20 x 10 <5> cells, suspended in 100 μΐ of culture medium, were incubated for 30 minutes at 4 ° C in the presence of

La sospensione cellulare fu quindi centrifugata per 5 min. a 2000 g, il pellet fu lavato e risospeso in terreno fresco. La centrifugazione e il lavaggio cellulare furono ripetuti 2 volte. L’entità della radiomarcatura fu valutata misurando la radioattività legata alle cellule (cpm/105 cellule) con un contatore gamma. The cell suspension was then centrifuged for 5 min. at 2000 g, the pellet was washed and resuspended in fresh medium. Centrifugation and cell washing were repeated 2 times. The extent of the radiolabelling was evaluated by measuring the radioactivity linked to the cells (cpm / 105 cells) with a gamma counter.

La specificità del legame fu determinata valutando lo spiazzamento radioattivo. La sospensione cellulare fu incubata per 5 minuti in presenza di LVT o di OT 100 μΜ e 1 mM, oppure in presenza di peptidi di controllo (quali somatostatina 100 nM e 1 μΜ ed esarelina 1 μΜ) seguita da 20 minuti di incubazione con Le cellule furono quindi centrifugate e lavate due volte come sopra riportato. L’attività dei due diversi agonisti e dei peptidi di controllo nel competere con il radioligando fu valutata misurando la radioattività legata alle cellule. Tutti gli esperimenti furono effettuati in triplicato. L’analisi statistica fu effettuata con ANOVA. Il limite di significatività era 0,05. Dopo 30 minuti di incubazione con limmunoconiugato marcato con indio secondo l’invenzione, si osservò la presenza di radiomarcatura specifica in tutte le linee cellulari OTR+ (MCF7, TS/A, MOG-U-V-W), che era invece trascurabile nelle cellule HT29 (OTR-). La quantità di marcatura, espressa come cpm/105 cellule, era 1711 ± 97 per le cellule MCF7, 7252 83 per le cellule TS/A e 1811 ± 101 per le cellule MOG-U-V-W, mentre era solo di 34 6 per le cellule HT29. La specificità di legame fu dimostrata per spiazzamento per radioligando freddo; nelle cellule OTR+, più del 90% del legame specifico fu spiazzato per pre-incubazione di 5 minuti con OT o LVT non radioattivo alle concentrazioni di 100 μΜ e 1 mM (figura 2). Un’ulteriore conferma della specificità di legame era fornita dalla mancanza di spiazzamento di radioligando in seguito ad incubazione con due peptidi di controllo, la somastotatina e l’ esarelina, impiegate a concentrazioni di 100 nM e 1 μΜ. The specificity of the binding was determined by evaluating the radioactive displacement. The cell suspension was incubated for 5 minutes in the presence of LVT or OT 100 μΜ and 1 mM, or in the presence of control peptides (such as somatostatin 100 nM and 1 μΜ and hexarelin 1 μΜ) followed by 20 minutes of incubation with the cells. they were then centrifuged and washed twice as reported above. The activity of the two different agonists and of the control peptides in competing with the radioligand was evaluated by measuring the radioactivity linked to the cells. All experiments were carried out in triplicate. Statistical analysis was carried out with ANOVA. The limit of significance was 0.05. After 30 minutes of incubation with the indium-labeled immunoconjugate according to the invention, the presence of specific radiolabelling was observed in all OTR + cell lines (MCF7, TS / A, MOG-U-V-W), which was negligible in HT29 cells (OTR- ). The amount of labeling, expressed as cpm / 105 cells, was 1711 ± 97 for MCF7 cells, 7252 83 for TS / A cells and 1811 ± 101 for MOG-U-V-W cells, while it was only 34 6 for HT29 cells . The specificity of binding was demonstrated by displacement by cold radioligand; in OTR + cells, more than 90% of the specific binding was displaced by pre-incubation of 5 minutes with non-radioactive OT or LVT at concentrations of 100 μΜ and 1 mM (Figure 2). Further confirmation of the specificity of binding was provided by the lack of displacement of the radioligand following incubation with two control peptides, somastotatin and hexarelin, used at concentrations of 100 nM and 1 μΜ.

ESEMPIO 5 - Studi in vivo. EXAMPLE 5 - In vivo studies.

Per determinare l’entità e la specificità della captazione mediata dal recettore di To determine the extent and specificity of the receptor-mediated uptake of

DOTA-LVT in tumori in paragone a un coniugato non specifico di controllo DOTA-LVT in tumors compared to a non-specific control conjugate

TOC) si è impiegato il modello sperimentale del tumore TS/A, OTR+ e SSTR- in topi Balb/c. Un totale di 8 topi femmina Balb/c del peso di 25 g, furono trattati per via sottocutanea con cellule di carcinoma mammario TS/A (1 x 10<6>, in 0,2 ml di terreno). 20 giorni più tardi, quando il tumore aveva raggiunto una dimensione di circa 2 cm di diametro, gli animali furono trattati per via intraperitoneale con una miscela di 1.1 MBq di e TOC) the experimental model of TS / A, OTR + and SSTR- tumor in Balb / c mice was employed. A total of 8 female Balb / c mice weighing 25 g were subcutaneously treated with TS / A breast cancer cells (1 x 10 <6>, in 0.2 ml of medium). 20 days later, when the tumor had reached a size of approximately 2 cm in diameter, the animals were treated intraperitoneally with a mixture of 1.1 MBq of e

Gli animali furono divisi in gruppi e sacrificati a 3 e 24 ore The animals were divided into groups and sacrificed at 3 and 24 hours

dopo il trattamento, rispettivamente. Il tumore, sangue, fegato, reni e cervello furono rimossi e pesati. La radioattività fu misurata in un rivelatore a raggi gamma assieme agli standard della miscela di iniezione a due diversi punti di tempo in modo da calcolare il contributo di ogni isotopo: immediatamente dopo la rimozione dei tessuti e dopo 2 settimane, corrispondenti a 5 emivite fisiche di L’attività fu espressa come percentuale di dosi iniettate per mg di tessuto e i rapporti di captazione fra tumore e cervello rispetto al sangue vennero calcolati. Si effettuò un test statistico (T-test) per determinare la differenza di significatività fra i gruppi. Il limite per la significatività era di 0,05. I risultati dello studio di biodistribuzione in vivo hanno dimostrato che i composti in esame non si accumulavano nel cervello. I rapporti di captazione del tumore rispetto al sangue e del cervello rispetto al sangue per entrambi i peptidi radiomarcati sono riportati nella tabella successiva: after treatment, respectively. The tumor, blood, liver, kidney and brain were removed and weighed. The radioactivity was measured in a gamma-ray detector together with the standards of the injection mixture at two different time points in order to calculate the contribution of each isotope: immediately after tissue removal and after 2 weeks, corresponding to 5 physical half-lives of Activity was expressed as a percentage of injected doses per mg of tissue, and tumor-brain-to-blood uptake ratios were calculated. A statistical test (T-test) was performed to determine the difference in significance between the groups. The limit for significance was 0.05. The results of the in vivo biodistribution study showed that the test compounds did not accumulate in the brain. Tumor versus blood and brain versus blood uptake ratios for both radiolabeled peptides are shown in the following table:

TABELLA TABLE

Rapporti di captazione: tumore/sangue e cervello/sangue Uptake ratios: tumor / blood and brain / blood

Claims (9)

RIVENDICAZIONI 1. Coniugati di sostanze capaci di legarsi ai recettori dell’ossitocina con agenti citotossici o radioattivi. CLAIMS 1. Conjugates of substances capable of binding to oxytocin receptors with cytotoxic or radioactive agents. 2. Coniugati secondo la rivendicazione 1 in cui l’analogo di ossitocina ha un residuo di amminoacido funzionalizzabile in posizione 8 della sequenza peptidica dell’ ossitocina. 2. Conjugates according to claim 1 in which the oxytocin analog has a functionalizable amino acid residue in position 8 of the peptide sequence of oxytocin. 3. Coniugati secondo la rivendicazione 2, in cui l’analogo di ossitocina è LVT (lisina- vasotocina) . 3. Conjugates according to claim 2, in which the oxytocin analog is LVT (lysine-vasotocin). 4. Coniugati secondo la rivendicazione 1, 2 o 3 in cui gli agenti citotossici o antiproliferativi sono anticorpi, tossine, farmaci citotossici, chelanti di metalli radioattivi o paramagnetici, biotina. 4. Conjugates according to claim 1, 2 or 3 wherein the cytotoxic or antiproliferative agents are antibodies, toxins, cytotoxic drugs, radioactive or paramagnetic metal chelators, biotin. 5. Coniugati secondo la rivendicazione 4 in cui gli agenti citotossici sono chelanti di metalli o alogeni radioattivi o di metalli paramagnetici. 5. Conjugates according to claim 4 wherein the cytotoxic agents are metal or radioactive halogen or paramagnetic metal chelators. 6. Coniugati secondo la rivendicazione 5 in cui i chelanti sono scelti fra DOTA, BOPTA, DOTMA. 6. Conjugates according to claim 5 wherein the chelators are selected from DOTA, BOPTA, DOTMA. 7. I coniugati della rivendicazione 6 chelati con metalli o alogeni radioattivi scelti 7. The conjugates of claim 6 chelated with selected radioactive metals or halogens 8 Coniugati della rivendicazione 7 come traccianti o agenti radioterapici per la localizzazione o il trattamento di tumori che esprimono OTR. 8 Conjugates of claim 7 as tracers or radiotherapeutic agents for the localization or treatment of tumors expressing OTR. 9. Composizioni farmaceutiche e diagnostiche contenenti quantità efficaci dei coniugati delle rivendicazioni 1-8. 9. Pharmaceutical and diagnostic compositions containing effective amounts of the conjugates of claims 1-8.
IT2001MI001167A 2001-06-01 2001-06-01 CYTOTOXIC OR RADIOACTIVE CONJUGATES CAPABLE OF BINDING TO OXYTOCIN RECEPTORS ITMI20011167A1 (en)

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PCT/EP2002/005687 WO2002098447A1 (en) 2001-06-01 2002-05-24 Cytotoxic or radioactive conjugates able to bind to oxytocin receptors

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