US20200085979A1

US20200085979A1 - Precision medicine theranostics and diagnostics a combination thereof

Info

Publication number: US20200085979A1
Application number: US16/568,227
Authority: US
Inventors: Stanley Satz
Original assignee: Individual
Current assignee: Individual
Priority date: 2018-09-11
Filing date: 2019-09-11
Publication date: 2020-03-19
Also published as: EP3923952A1; US11357874B2; EP3923952A4; WO2020056064A1; US20200085981A1; JP2022500464A

Abstract

The component ‘drug’ new molecule for combining metals into complexes through a ring structure (DOTA) or linear structure, and a radionuclidic component, and chelating agent wherein embodiments may include a companion diagnostic, and in which embodiments further include anti-integrin precision medicines for cancers expressing αvβ3 and αvβ5 integrins, second component for combining metals into complexes through a ring or linear structure, and one or more radionuclidic components, a chelating agent for diagnosis and/or therapy (theranostic) in which the embodiments further include a anti-integrin peptidomimetic for precision medicine for cancers expressing αvβ3 and αvβ5 integrins.

Description

CROSS-REFERENCE TO RELATED APPLICATION

This application claims the benefit of priority to U.S. Provisional Application Ser. No. 62/729,959, filed on Sep. 11, 2018, the complete contents of which are herein incorporated by reference.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention relates to the field of cancer diagnosis and therapy, theranostics, and the creation of novel molecules; more particularly an integrin peptidomimetic, an isotope and a chelating agent for treating, diagnosing and staging cancers, and, in particular, cancers overexpressing the human epidermal growth factor positive receptor 2 protein (HER2+) and triple negative breast cancer and relates to the synthesis and reaction of potent and selective small molecule integrin antagonists containing appropriate linkers and functional groups for chemical reaction with other molecules which contain reactive nucleophiles such as thiols such that a covalent linkage is formed between a moiety to be conjugated and the targeting entity. The small molecule targeting antagonist anti αvβ3 binds to cognate receptor systems with high specificity. The covalently linked moiety includes peptidometics preferably but may also be small molecule therapeutics, nanoparticles, polymers, peptides, oligonucleotides, and antineoplastics.

2. Brief Description of the Related Art

Integrins are transmembrane receptors that facilitate cell-extracellular matrix (ECM) adhesion. Upon ligand binding, integrins activate signal transduction pathways that mediate cellular signals such as regulation of the cell cycle, organization of the intracellular cytoskeleton, and movement of new receptors to the cell membrane. The presence of integrins allows rapid and flexible responses to events at the cell surface (e.g. signal platelets to initiate an interaction with coagulation factors).
Integrins are obligate heterodimers, meaning that they have two subunits: α (alpha) and β (beta). Integrins in mammals have eighteen α and eight β subunits, in Drosophila five α and two β subunits, and in Caenorhabditis nematodes two α subunits and one β subunit. The α and β subunits each penetrate the plasma membrane and possess several cytoplasmic domains.
Integrins can be categorized in multiple ways. For example, some a chains have an additional structural element (or “domain”) inserted toward the N-terminal, the alpha-A domain (so called because it has a similar structure to the A-domains found in the protein von Willebrand factor; it is also termed the α-I domains.
Integrins carrying this domain either bind to collagen (e.g. integrins α1 β1, and α2 β1), or act as cell-cell adhesion molecules (integrins of the β2 family). This α-I domain is the binding site for ligands of such integrins. Those integrins that don't carry this inserted domain also have an A-domain in their ligand binding site, but this A-domain is found on the β subunit.
Integrins play an important role in cell signaling by modulating the cell signaling pathways of transmembrane protein kinases such as receptor tyrosine kinases (RTK). While the interaction between integrin and receptor tyrosine kinases originally was thought of as unidirectional and supportive, recent studies indicate that integrins have additional, multi-faceted roles in cell signaling. Integrins can regulate the receptor tyrosine kinase signaling by recruiting specific adaptors to the plasma membrane. For example, β1c integrin recruits Gab1/Shp2 and presents Shp2 to IGF1R, resulting in dephosphorylation of the receptor. In the reverse direction, when a receptor tyrosine kinase is activated, integrins co-localise at focal adhesion with the receptor tyrosine kinases and their associated signaling molecules.
The αvβ3 integrin is a receptor for vitronectin [Hermann, P. et al. “The vitronectin receptor and its associated CD47 molecule mediates proinflammatory cytokine synthesis in human monocytes by interaction with soluble CD23” [The Journal of cell biology 144 (1999): 767-75]. It consists of two components, integrin αv and β3 (CD61), and is expressed by platelets as well as other cell types. It has been shown that inhibitors of αvβ3 like etaracizumab may be used as antiangiogenics.
Integrins are members of a family of heterodimeric transmembrane cell surface receptors that play a crucial role in cell-cell and cell-matrix adhesion processes (Hynes, R. O. Cell 1992, 69, 11-25). These receptors consist of an αvβ3 and αvβ5 which non-covalently associate in defined combinations (Eble, J. A. Integrin-Ligand Interaction; Springer: Heidelberg, 1997; pp 1-40). Most of them recognize the Arg-Gly-Asp (RGD) triad found in many extracellular matrix proteins (i.e., vitronectin), Serini, G.; et al. A sticky business. Exp. Cell. Res. 2005, and snake venom disintegrins (Ruoslahti, E.; Pierschbacher, M. Cell 1986, 44, 517-518; D'Souza, S. E.; et al. Trends Biochem. Sci. 1991, 16, 246-250; Gould, R. J.; et al. Proc. Soc. Exp. Biol. Med. 1990, 195, 168-171). Even if different integrins recognize different proteins containing the RGD sequence, several studies have demonstrated that the amino acid residues flanking the RGD sequence of high-affinity ligands modulate their specificity of interaction with integrin complexes. Despite numerous studies reported in the literature, ligand selectivity toward different integrin subtypes is still a challenging problem mainly because most of the 3D-structures of integrin subtypes remain unknown (Marinelli, L.; et al. J. Med. Chem. 2004, 47, 4166-4177).
The αv⊖3.integrin is strongly expressed on activated endothelial and melanoma cells and in contrast, it is weakly expressed in quiescent blood vessels and pre-neoplastic melanomas (Hood, J. D.; Cheresh, D. A. Nat. Rev. Cancer 2002, 2, 91-100). Along with αvβ3 beta.sub.5 integrin, .AVB3 is reported to be involved in physiological processes including angiogenesis and tissue repair as well as pathological conditions such as tumor induced angiogenesis (Eliceiri, B. P.; Cheresh, D. A. J. Clin. Invest. 1999, 103, 1227-1230; Kumar, C. C. Curr. Drug Targets 2003, 4, 123-131), tumor cell migration and invasion (Clezardin, P. Cell. Mol. Life. Sci. 1998, 54, 541-548). Despite the fact that both integrins promote cell attachment to vitronectin and participate in the same processes, they are reported to be structurally designed to respond to different signaling events. Previous studies provided evidence that bFGF-induced angiogenesis is mediated by integrin αvβ3.whereas VEGF-induced angiogenesis is mediated by αvβ3 beta sub.5 (Friedlander, M.; et al. Science 1995, 270, 1500-1502). Melanoma cells expressing integrin αvβ3.migrate in vitro and metastasize in vivo without the need for exogenous cytokine stimulation (Filardo, E. J.; et al. J Cell Biol. 1995, 130, 441-450). Conversely, tumor cells expressing αvβ3 beta.sub.5 integrin require a tyrosine kinase receptor-mediated signaling event for motility on vitronectin and in vivo dissemination (Brooks, P. C.; et al J Clin Invest. 1997, 99, 1390-1398). While αvβ3 beta.sub.5 is widely expressed by many malignant tumor cells, .AVB3 has a relatively limited cellular distribution compared with that of αvβ3 beta.sub.5 (Pasqualini, R.; et al. J. Cell Science 1993, 105, 101-111; Walton, H. L.; et al. J. Cell. Biochem. 2000, 78, 674-680.). Therefore, in order to target αvβ3 beta sub.3-mediated processes for diagnostic or therapeutic purposes, the development of new compounds that can discriminate between .ABV3 and αvβ3 beta.sub.5 is required.
αvβ3 integrin interacts with HER2+ in cancer cells and can regulate HER2+ localization. The combined impacts of αv-integrin and HER2+ influence the invasive phenotype of breast cancer cells. Targeting αv-integrin in HER2+ breast cancer may slow growth and decrease infiltration in the normal brain (Lal 2015).
A need remains for further improved targeting of integrins; as well as substances such as small molecules, peptides, other nucleic acids, fluorescent moieties, polymers, alpha, beta or alpha/beta tandem radionuclides to particularly target cancer cells, destroy vascular endothelial growth factor—new blood vessels that feed tumors, target the cytoplasm of such cells and impair the DNA of cancer cells.

SUMMARY OF THE INVENTION

The invention relates to compositions for treating HER2+ cancers and cancers overexpressing the human epidermal growth factor positive receptor 2 protein (HER2+) and triple negative breast cancer, and includes pharmaceutical preparations and the compositions in a carrier or other or excipient for delivery to a patient. Diagnostic compositions also are provided and are used for detecting the presence of a tumor in a patient. The invention further relates to the synthesis and reaction of potent and selective small molecule integrin antagonists containing appropriate linkers and functional groups for chemical reaction with other molecules which contain reactive nucleophiles such as thiols such that a covalent linkage is formed between a moiety to be conjugated and the targeting entity. The small molecule targeting antagonist anti αvβ3 binds to cognate receptor systems with high specificity. The covalently linked moiety includes peptidometics preferably but may also be small molecule therapeutics, nanoparticles, polymers, peptides, oligonucleotides, and antineoplastics.
Preferred embodiments of the compositions include a peptide and more particularly a peptidomimetic, linked to a spacer and chelated to a radionuclide, wherein the peptide comprises an αvβ3 integrin antagonist.
More particularly, the invention relates to compounds of a formula comprising an αvβ3 integrin antagonist, and more particularly a peptidomimetic, which is radiolabeled with a radionuclide. According to preferred embodiments the radionuclide is [177]Lu or [225]Actinium; and according to other embodiments the radionuclide may be Bismuth-213, Thorium-227, Lead-212, Astatine-211, Yttrium-90, Iodine-131, Gallium-68, Zirconium-68 or other therapeutic radiolabel.
Embodiments of the invention involve the following therapeutic compositions: [177Lu] DOTAGA IAC or a ligand containing the active moiety, the αvβ3 integrin antagonist peptidomimetic, 4-[2-(3,4,5,6-tetrahydropyrimidine-2-ylamino) ethyloxy]benzoyl-2[N-(3-aminoneopenta-1-carbamyl)]-aminoethylsulfonyl-amino-β-alanine (IAC) and 1-(1-carboxy-3-carbotertbutoxymethyl)-1,4,7,10-tetraazacyclododecane (DOTAGA) (tBu)4; or 4-[4,7-Bis-(carboxymethyl)-[1,4,7]triazonan-1-yl]-4-carboxy-butyryl (NODAGA); DOTATATE, or C26H34N4O10S (CHX-A), radiolabeled with Lutetium-177, Actinium-225, Bismuth-213, Thorium-227, Lead-212, Astatine-211, Yttrium-90, Iodine-131, Gallium-68 and Zirconium-68 or a combination of radionuclides for tumor targeted radionuclide therapy (TRNT); and of the formula Ga-68-NODAGA IAC or a ligand containing the active moiety, the αvβ3 integrin antagonist peptidomimetic 4-[2-(3,4,5,6-tetrahydropyrimidine-2-ylamino)ethyloxy]benzoyl-2-[N-(3-aminoneopenta-1-carbamyl)]-aminoethylsulfonyl-amino-β-alanine (IAC) and to 1-(1-carboxy-3-carbo-t-butoxypropyl)-4,7-(carbo-tert-butoxymethyl)-1,4,7-triazacyclononane. The former composition is preferably used for therapeutic treatment, while the latter composition (i.e., Ga-68 composition) is used for diagnostic imaging. The former and the latter compositions are companion compositions so that the therapeutic administration of the former compound (to a patient) may be carried out and imaging of the applied therapeutic results (to detect cancer) may be carried out using the latter Ga-68 composition (which also is administered to a patient prior to or in connection with imaging).
The αvβ3/αvβ5 anti-integrin antagonist peptidomimetic can be used for the diagnosis and treatment of cancers, and comprises a pharmaceutical composition consisting of a therapeutically effective amount of the αvβ3 and/or αvβ5 antagonist peptidomimetic compound, covalently linked by a spacer sequence wherein the compound may be a linear artificial sequence, a chelator such as NODAGA, DOTAGA, DOTATATE, or CHX-A, and a pharmaceutically acceptable excipient for tumor targeted radionuclide therapy (TRNT) using a radionuclide such as Lutetium-177, Actinium-225, Bismuth-213, Thorium-227, Lead-212, Astatine-211, Yttrium-90, Iodine-131, Gallium-68, Zirconium-68, or other therapeutic radiolabel.
The compounds of this invention contain an αvβ3 and αvβ5 anti-integrin antagonist peptidomimetic and unlike cyclical peptides display highly selective affinity for the HER2+ protein, vascular endothelial growth factor, vitronectin, fibronectin, tenascin, reelin, kindlin, talin or combinations thereof for the diagnosis or prognosis of disorders using PET, SPECT and MRI by administering an effective amount of αvβ3/αvβ5 anti-integrin antagonist peptidomimetic thereof to a subject, particularly for the diagnosis of diseases related to metastasis and angiogenesis such as breast cancer, lung cancer, ovarian cancer, gastric cancer, esophageal cancer, blood borne cancers, musculoskeletal tumors, melanoma, head and neck cancer, human glioma, vascular restenosis, osteoporosis, rheumatoid arthritis. The aforementioned compound demonstrates safety and efficacy in animal and mammalian models.
Diagnostic kits or synthesizer cartridges are provided comprising the αvβ3 and αvβ5 peptidomimetic antagonist attached to a chelator such as NODAGA, DOTAGA, DOTATATE or CHX-A or a nanocarrier such as a gold nanoparticle for disease detection, staging, non-invasive determination of treatment efficacy which may be in tissue, bone or in plasma for treatment of pathologies, such as breast cancer, esophageal cancer, gastric cancer, lung cancers, ovarian cancer, pancreatic cancer, musculoskeletal tumors, melanomas, blastomas, sarcomas, cytomas, carcinomas, rhabdoid tumors, head and neck cancer, human glioma, cervical cancer, vascular restenosis, osteoporosis, rheumatoid arthritis and other indications.
In specific embodiments, a tumor targeted radionuclidic agent contains a [68Ga], [18F], [203Pb], [99mTc] or [89Zr] as a diagnostic. For tumor targeted therapy the radionuclide may be [177]Lu, [225 Ac], [90Y], [227Th], more particularly, a [177Lu] DOTAGA IAC or a ligand containing the active moiety, the αvβ3 integrin antagonist peptidomimetic, 4-[2-(3,4,5,6-tetrahydropyrimidine-2-ylamino) ethyloxy]benzoyl-2-[N-(3-aminoneopenta-1-carbamyl)]-aminoethylsulfonyl-amino-β-alanine (IAC) and 1-(1-carboxy-3-carbotertbutoxymethyl)-1,4,7,10-tetraazacyclododecane (DOTAGA) (tBu)4, or 4-[4,7-Bis-(carboxymethyl)-[1,4,7]triazonan-1-yl]-4-carboxy-butyryl (NODAGA), or C26H34N4O10S (CHX-A), to radiolabel with Lutetium-177, Actinium-225, Bismuth-213, Thorium-227, Lead-212, Astatine-211, Yttrium-90, Iodine-131, Gallium-68, Zirconium-68, or a combination of radionuclides for tumor targeted radionuclide therapy (TRNT).
The peptidomimetic compositions may comprise first and second binding regions, including a first region that is an antigen binding region and a second region that is an integrin binding region. The second integrin binding region binds an epitope to a paratope, and the first antigen-binding region binds an epitope on human epidermal growth factor receptor 2 (HER2) and blocks the binding to HER2, optionally soluble HER2, wherein the peptidomimetic comprises an anti-integrin that shows high affinity for integrin αvβ3/αvβ3, vascular endothelial growth factor, vitronectin, fibronectin and tenascin, actin, reelin, talin or kindlin.
Compositions of the invention comprise integrin receptor peptidomimetic antagonists whose molecular structure includes a tetrahydropyridimidinyl aminoethyloxybenzoyl group on a sulfonylamino-β-alanine nucleus exhibit increased binding affinity for the αvβ3/αvβ3 receptor when further substituted on the sulfonyl moiety with an N-amino alkycarbamyl group or a butyloxycarbonylamino alkylcarbamoyl group or similar groups.
In particular, the present invention relates to the compounds of formulas herein for the improved delivery of conjugated moieties such as small molecules, peptidomimetics, peptides, nucleic acids, fluorescent moieties, antineoplastics, nanoparticles and polymers to target cells expressing the .αvβ3 dimer for various therapeutic and other applications. The present invention also relates to methods of manufacturing and using such compounds.
Integrin receptor antagonists whose molecular structure includes a tetrahydropyridimidinyl aminoethyloxybenzoyl group on a sulfonylamino-β-alanine nucleus exhibit increased binding affinity for the αvβ3 receptor when further substituted on the sulfonyl moiety with an N-amino alkycarbamyl group or a butyloxycarbonylamino alkylcarbamoyl group or similar groups. Embodiments of the present invention also utilize the therapeutic compounds of the invention in conjunction with a diagnostic. Compounds containing αvβ3/αvβ5 anti-integrin antagonist peptidomimetic, displaying a selective affinity for the HER2+ protein, vitronectin, fibronectin, tenascin, reelin, kindlin, talin or combination thereof for the diagnosis or prognosis of disorders using PET, SPECT and MRI or combinations by administering an effective amount of activity thereof to a subject, particularly for the diagnosis of diseases related to metastasis and angiogenesis such as breast cancer, lung cancer, ovarian cancer, gastric cancer, esophageal cancer, blood borne cancers, musculoskeletal tumors, melanoma, head and neck cancer, human glioma, cervical cancer, vascular restenosis, osteoporosis, rheumatoid arthritis. The aforementioned compound demonstrates safety and efficacy theranostically in animal and human mammalian models. The compounds referred to may be radiotracers for nuclear medicine, such as the Gallium-68 or Fluorine-18, Zirconium-68, Copper-64, or Technetium-99m, covalently bound to a spacer, chelator, directly to the peptidomimetic, to a nanoparticle or to more than one amino acid unit of the peptidomimetic. Other radiotracers are those reported in the publication Radionuclide Peptide Cancer Therapy, edited by M. Chinol and G. Paganelli, Taylor and Francis CRC Press, 2006. Studies have been performed comparing this cyclical αvβ3 and αvβ5 integrin antagonist peptidomimetic to a linear αvβ3 integrin peptide in nude mice using the same chelator and radionuclide. The results for this αvβ3 and αvβ5 linear peptidomimetic were three to seven times better affinity for the targeted organs. (See LinnebacHER2+M1, Maletzki C. “Tumor-infiltrating B cells: The ignored players in tumor immunology.” Oncoimmunology. 2012 Oct. 1; 1 (7):1 186-1 188. 15. Nelson B H. CD20B cells: the other tumor-infiltrating lymphocytes. J lmmunol. 2010 Nov. 1; 185(9):4977-82.) In human studies comparing this cyclical αvβ3 and αvβ5 integrin antagonist peptidomimetic to a ‘gold standard’ [18F] Fludeoxyglucose PET scan in an HER2+ breast cancer patient, the integrin antagonist peptidomimetic enabled detection of 25 tumors distant from the primary versus only 12 with [18F] Fludeoxyglucose. Detection may be effected using a compound containing the αvβ3 and αvβ5 integrin antagonist peptidomimetic, displaying a selective affinity for the HER2+ protein, vitronectin, fibronectin, tenascin, reelin, kindlin, talin or combination thereof, and may comprise a pharmaceutical composition comprising a therapeutically effective amount of the αvβ3 and αvβ5 antagonist peptidomimetic compound, covalently linked by a spacer sequence, wherein the compound may be a linear artificial sequence, a chelator such as NODAGA, DOTAGA, CHX-A and a pharmaceutically acceptable excipient for targeted radionuclide therapy (TRNT). The compound may also be a radiotracer for nuclear medicine, such as the Gallium-68 or Fluorine-18, Zirconium-68, Copper-64, or Technetium-99m, covalently bound to a spacer, chelator, directly to the peptidomimetic, to a nanoparticle or to more than one amino acid unit of the peptidomimetic. Other radiotracers are those reported in the publication Radionuclide Peptide Cancer Therapy, edited by M. Chinol and G. Paganelli, Taylor and Francis CRC Press, 2006.
The peptide hereinafter referred to as integrin which may be a bifunctional molecule containing a linear RGD motif and a sequence corresponding to a C-terminal connected by a linker.

Synthesis and Production of Compounds

This peptidomimetic may be synthesized by a suitable production method, one such exemplary method being the solid-phase method using Fmoc chemistry. Amino acids are coupled according to the HBTU/HOBt/DIPEA procedure (Fields, C. G.; et al Pept. Res. 1991, 4, 95-101). Final deprotection and cleavage from the resin were achieved with TFA and scavengers. The purity and the identity of the peptides are confirmed by analytical RP-HPLC and MALDI-TOF mass spectrometry. Peptidomimetics are synthesized on an automated peptide synthesizer using Fmoc solid-phase strategy (0.25 mmol).
According to a preferred implementation, the α5β3 integrin peptide mimics that bind α5β3 integrin, may be produced by culturing a host cell so that the anti-α5β3 integrin antibody or antibody is produced. The composition may be attached to a suitable spacer sequence and/or a chelator, the radionuclide being attached to or via the chelator.

DETAILED DESCRIPTION OF THE INVENTION

Unless otherwise indicated, the following specific terms and phrases used in the description and claims are defined as follows:
The term “moiety” refers to an atom or group of chemically bonded atoms that is attached to another atom or molecule by one or more chemical bonds thereby forming part of a molecule.
The term “conjugated moiety” refers to moiety which is a therapeutic or useful compound, peptidomimetic, peptide, polymer, small molecule, fluorescent moiety, oligonucleotide or nucleic acid. Examples include drugs, diagnostic peptidomimetics, therapeutic peptidomimetics, therapeutic peptides, antisense oligonucleotides, siRNA, and fluorescein isothiocyanate (FITC).
The term “alkyl” denotes a monovalent linear or branched saturated hydrocarbon group of 1 to 12 carbon atoms. In particular embodiments, alkyl has 1 to 7 carbon atoms, and in more particular embodiments 1 to 4 carbon atoms. Examples of alkyl include methyl, ethyl, propyl, isopropyl, n-butyl, iso-butyl, sec-butyl, or tert-butyl.

Tumor Targeted Radionuclide Therapy (TRNT)

An objective of TRNT is to selectively deliver cytotoxic radiation to cancer cells causing minimal toxicity to surrounding healthy tissues. This is achieved by delivering a radioactive payload into the tumor cells. Examples of this approach are Gallium-68 (68Ga), Yttrium-90 (90Y), Lutetium-177 (177Lu), Iodine-131, (177Lu), Zirconium-68, Actinium-225 (225Ac), Bismuth-213 (213Bi) and Lead-212 (212Pb) as radiolabeled peptides to treat tumors overexpressing specific proteins. In addition, monoclonal antibodies (abs) are also used as vehicles to target tumor-associated antigens providing internal radiotherapy. Regulatory-agency approved radiolabeled agents are 90Y-ibritumomab and 90I, and 177Lu Lutathera for treatment of gastroenteropancreatic tumors. Thus, a therapeutic radiopharmaceutical can consist of two parts: a targeting biomolecule that specifically determines the localization of the radiopharmaceutical and a radionuclide that delivers the mechanism of action through its decay. Radiopharmaceuticals are used as diagnostics for non-invasive imaging by detection of γ-rays using positron emission tomography (PET) or single-photon emission computerized tomography (SPECT), and/or as therapeutics for TRNT to deliver cancer cell killing radiation to the targeted tumor cells.
The crucial roles of integrin αvβ3 in tumor angiogenesis have led to a promising strategy to block signaling by antagonists, as this would theoretically inhibit tumor angiogenesis or enhance the efficacy of other tumor therapeutics. In addition, the high expression of integrin αvβ3 on tumor new-blood vessels and some tumor cells makes the anti-integrin αvβ3 suitable marker for cancer-targeted drug delivery. According to a preferred embodiment, the therapeutic compound comprises Ga-68 DOTAGA and Lu-177 DOTAGA: A ligand containing the active moiety, the αvβ3 integrin antagonist peptidomimetic, 4-[2-(3,4,5,6-tetrahydropyrimidine-2-ylamino) ethyloxy]benzoyl-2-[N-(3-aminoneopenta-1-carbamyl)]-aminoethylsulfonyl-amino-β-alanine (IAC) and 1-(1-carboxy-3-carbon tert butoxymethyl)-1,4,7,10-tetraazacyclododecane (DOTAGA) (tBu)4 to radiolabel Lutetium-177 for peptide receptor Lutetium therapy.
The preferred molecule used for tumor targeted radiotherapy in the invention consists of an integrin antagonist peptidomimetic isotope preferably [177]Lutetium.
According to a preferred embodiment, a compound of the invention is comprised of three components. The first component is the integrin antagonist peptidomimetic. This component is the peptidomimetic that targets the HER2+ cells. The second component is a chelator, a compound able to combine radiometals (such as Lu-177) into complexes through a ring-like structure; and the third optional component is the radionuclide to enhance uptake to the tumor and extend biological half life.
A potent αvβ3 integrin antagonist peptide mimic targeting agent specific for the HER2+ protein was developed, modified for use in humans and radiolabeled with Gallium-68, Fluorine-18 for diagnosis using positron emission tomography, or Lutetium-177 or Actinium-225 for use in humans enabling cancerous tumor detection using positron emission tomography and for tumor targeted radionuclide therapy (TRNT) alone or in combination with standard-of-care; immunotherapy and chemotherapy in the neoadjuvant setting. The radioligands enable non-invasive diagnosis and treatment of a wide range of cancers, while also reducing harmful side effects such as cardiovascular disease, peripheral neuropathy, and improving survival and quality of life.
The methods and therapeutic compositions of the present invention may be used in connection with a diagnostic. The diagnostic may be administered to the patient to identify areas of tumor or cancer cells or growth, and may be used to determine one or more subsequent treatment parameters for a treatment protocol, such as the treatment dosage and extent (e.g., where the treatment involves administering an a treatment compound according to the invention, such as the preferred composition, Lu-177 DOTAGA IAC. According to some embodiments, a preferred diagnostic composition may comprise Ga-68-NODAGA IAC: A ligand containing the active moiety, the αvβ3 integrin antagonist peptidomimetic 4-[2-(3,4,5,6-tetrahydropyrimidine-2-ylamino)ethyloxy]benzoyl-2-[N-(3-aminoneopenta-1-carbamyl)]-aminoethylsulfonyl-amino-β-alanine (IAC) and attached (e.g., chelated) to 1-(1-carboxy-3-carbo-t-butoxypropyl)-4,7-(carbo-tert-butoxymethyl)-1,4,7-triazacyclononane.
The diagnostic and the therapeutic compositions preferably are prepared from components of a kit, where the composition is prepared and radiolabeled with the respective radionuclide prior to administering the composition to a patient/subject, which preferably is done intravenously.

PROPOSED EXAMPLES

Proposed Example 1

Example 1: a preferred compound of the invention, comprising Lu-177 DOTAGA IAC, is administered to a human patient at a fixed dose of 7.4 GBq (every 12 weeks) up to a cumulative dose which is tolerated by the patient (maximum 29.6 GBq). The preferred compound preferably is administered intravenously. Nivolumab is administered twice for each treatment of the preferred Lu-177 DOTAGA IAC compound, at a dose of 3 mg/Kg: one administration seven days before (d−7) and the other administration seven days after (d−positive7) administration of the compound of the invention, with the aim of achieving an effective PD-1/PD-L1 blockade, but also in the need not to overlap the anticipated lymphocyte nadir related to the lymphocytopenia-induced effect of the compound of the invention.
Studies have shown that intravenous administration of amino acids has a renal protective effect. An infusion of amino acids (containing lysine and arginine) could be done 30 to 45 minutes before the administration of 177Lu-DOTATATE IAC and last for 3 to 4 hours.

Proposed Example 2

Example 2: In a proposed example, the compound of the present invention was administered to a patient suffering from HER2+ breast cancer. The patient was given the compound of the invention (e.g., in an amount of from about 3.7-7.4 GBq) administered intravenously, and representing a dosage. One or more, and preferably a plurality of subsequent treatments of a similar amount are dosed to the patient intravenously, a couple to a few weeks from the first dosage. In this example, an additional therapeutic may be administered on either side of a window based on when the patient receives the inventive compound doses, e.g., such as seven days prior to a dose and seven days after a dose.

Proposed Example 3

Example 3: The patient was treated as in proposed example 1, above, however, prior to treatment with the Lu-177 DOTAGA IAC, the patient was diagnosed using the preferred diagnostic composition, comprising Ga-68-NODAGA IAC.

Proposed Example 4

Example 4: The patient was treated as in proposed example 2, above, however, prior to treatment with the Lu-177 DOTAGA IAC, the patient was diagnosed using the preferred diagnostic composition, comprising Ga-68-NODAGA IAC.

Proposed Example 5

Example 5: The patient was treated as in Example 1, but with the radionuclide consisting of one or more of: Actinium-225, Bismuth-213, Thorium-227, Lead-212, Astatine-211, Yttrium-90, and Iodine-131, Gallium-68, and Zirconium-68.

Proposed Example 6

Example 6: The patient was treated as in Example 5, but with the chelator comprising one or more of: 4-[2-(3,4,5,6-tetrahydropyrimidine-2-ylamino) ethyloxy]benzoyl-2-[N-(3-aminoneopenta-1-carbamyl)]-aminoethylsulfonyl-amino-β-alanine (IAC), and (ii) at least one of the following:

- (a) 1-(1-carboxy-3-carbotertbutoxymethyl)-1,4,7,10-tetraazacyclododecane (DOTAGA) (tBu).sub.4; or
- (b) 4-[4,7-Bis-(carboxymethyl)-[1,4,7]triazonan-1-yl]-4-carboxy-butyryl (NODAGA); or
- (c) DOTATATE; or
- (d) C26H34N4O10S (CHX-A).

Claims

What is claimed is:

1. A therapeutic and diagnostic pharmaceutical composition for targeting and treating human epidermal growth factor receptor 2-Positive, hereinafter referred to as HER2+, or other cancer that overexpresses integrin receptors, the composition comprising an αvβ3/αvβ5 anti-integrin antagonist peptidomimetic that includes an integrin whose molecular structure includes a tetrahydropyridimidinyl-aminoethyloxybenzoyl group on a sulfonylamino-β-alanine nucleus, exhibiting selective, high binding affinity for α5β3; at least one radionuclide; a spacer sequence, wherein said integrin is covalently linked by a spacer sequence; a chelator selected from the group consisting of NODAGA, DOTAGA, DOTATATE, and CHX-A; and a pharmaceutically acceptable excipient.

2. The therapeutic and diagnostic composition of claim 1, wherein said spacer sequence is a linear artificial sequence.

3. The therapeutic and diagnostic composition of claim 1, wherein the radionuclide is selected from the group consisting of Lutetium-177, Actinium-225, Bismuth-213, Thorium-227, Lead-212, Astatine-211, Yttrium-90, Gallium-68, Zirconium-69 and Iodine-131.

4. The therapeutic composition of claim 1, wherein the radionuclide is selected from the group consisting of Lutetium-177.

5. The therapeutic composition of claim 3, wherein the radionuclide is selected from the group consisting of Actinium-225.

6. The therapeutic composition of claim 1, wherein said radionuclide is Lu-177, and wherein said chelator is DOTAGA, the active moiety consisting of: [177Lu] DOTAGA IAC.

7. The therapeutic composition of claim 1, wherein said spacer sequence is provided between said chelator and said tetrahydropyridimidinyl-aminoethyloxybenzoyl group on the sulfonylamino-β-alanine nucleus, exhibiting selective, high binding affinity for α5β3.

8. The therapeutic and diagnostic composition of claim 1, wherein said chelator is DOTAGA.

9. The therapeutic and diagnostic composition of claim 1, wherein said chelator is NODAGA.

10. The therapeutic and diagnostic composition of claim 1, wherein said chelator is DOTATATE.

11. The therapeutic and diagnostic composition of claim 1, wherein said chelator is CHX-A.

12. The therapeutic and diagnostic composition of claim 1, wherein said chelator also includes IAC.

13. A composition for tumor targeted radionuclide therapy (TRNT) for targeting and treating human epidermal growth factor receptor 2-Positive (HER2+) or other cancer that overexpresses integrin receptors, the composition comprising:

(1) a ligand containing the active moiety, said ligand comprising an αvβ3 integrin antagonist peptidomimetic, and

(i) 4-[2-(3,4,5,6-tetrahydropyrimidine-2-ylamino) ethyloxy]benzoyl-2-[N-(3-aminoneopenta-1-carbamyl)]-aminoethylsulfonyl-amino-β-alanine (IAC), and

(ii) at least one of the following:

(a) 1-(1-carboxy-3-carbotertbutoxymethyl)-1,4,7,10-tetraazacyclododecane (DOTAGA) (tBu).sub.4; or

(b) 4-[4,7-Bis-(carboxymethyl)-[1,4,7]triazonan-1-yl]-4-carboxy-butyryl (NODAGA); or

(c) DOTATATE;

(d) C26H34N4O10S (CHX-A); and

(2) a radionuclide selected from the group consisting of Lutetium-177, Actinium-225, Bismuth-213, Thorium-227, Lead-212, Astatine-211, Yttrium-90, Iodine-131, Gallium-68, Zirconium-68, or a combination of radionuclides.

14. The composition of claim 13, having a binding region with binding affinity for α5β3, and including a tetrahydropyridimidinyl-aminoethyloxybenzoyl group on a sulfonylamino-β-alanine nucleus.

15. The composition of claim 13, including a spacer sequence covalently linking said αvβ3 integrin antagonist peptidomimetic to said at least one of (ii)(a), or (ii)(b), or (ii)(c), or (iii)(d).

16. The composition of claim 14, wherein said composition comprises [177Lu] DOTAGA IAC.

17. A composition for tumor targeted radionuclide therapy (TRNT) for targeting and treating human epidermal growth factor receptor 2-Positive (HER2+) or other cancer that overexpresses integrin receptors, the composition comprising:

a) an αvβ3 integrin antagonist peptidomimetic; and

b) a radionuclide;

c) wherein said radionuclide is covalently bound to a spacer, chelator, directly to the peptidomimetic, to a nanoparticle or to more than one amino acid unit of the peptidomimetic; and

d) wherein said composition comprises one or more of the following to which said radionuclide is chelated:

(ii) at least one of the following:

(iii) 1-(1-carboxy-3-carbotertbutoxymethyl)-1,4,7,10-tetraazacyclododecane (DOTAGA) (tBu).sub.4; or

(iv) 4-[4,7-Bis-(carboxymethyl)-[1,4,7]triazonan-1-yl]-4-carboxy-butyryl (NODAGA); or

(v) DOTATATE; or

(vi) C26H34N4O10S (CHX-A).

18. The therapeutic composition of claim 1, wherein said integrin antagonist is a peptide mimetic chelated to a radionuclide.

19. The therapeutic composition of claim 1, containing the active moiety 4-[2-(3,4,5,6-tetrahydropyrimidine-2-ylamino) ethyloxy]benzoyl-2-[N-(3-aminoneopenta-1-carbamyl)]-aminoethylsulfonyl-amino-β-alanine (IAC).

20. The therapeutic composition of claim 1, containing the active moiety acetylsulfanyl-ethoxy}-ethoxy)ethox-y]ethoxy}-propionylamino)-propoxy]-benzyl}-ureido]-thiazole-4-carbonyl)-amino}-acetylamino]-phenyl-3-yl-propionic acid.