Conjugates of therapeutic or cytotoxic agents and biologically active peptides

Abstract

The invention features conjugates of therapeutic or cytotoxic agents and biologically active peptides and methods of use thereof.

Description

FIELD OF THE INVENTION[0001]The invention relates to conjugates of therapeutic or cytotoxic agents and biologically active peptides and uses thereof.BACKGROUND OF THE INVENTION[0002]The use of carbamate compounds as prodrugs is well known. Carbamate compounds are well suited for prodrug design because they can be used to regenerate the parent drug whether the point of connection to a vector molecule is a hydroxyl group or an amine. Carbamate prodrugs based on intramolecular cyclizations have been reported since the late 1980s. For example, U.S. Pat. No. 4,812,590 (which corresponds to EP 0 296 811) discloses derivatives of 4-hydroxyanisole carbamate as prodrugs for the delivery and concentration of 4-hydroxyanisole in melanomas. Vigroux et al. (J. Med. Chem. 38, 3983-3994, 1995) disclose prodrugs of acetaminophen which incorporate N-(substituted 2-hydroxyphenyl)- and N-(substituted 2-hydroxypropyl)carbamates.[0003]An optimal prodrug design is one in which the drug is conjugated to a...

AI Technical Summary

Benefits of technology

[0011]The invention features conjugates of a therapeutic agent or a cytotoxic agent and a targeting moiety. In particular, the invention features conjugates having a cleavable chemical linker which reduces the release of therapeutic or cytotoxic agent in circulation, thereby rendering the active agent component of the conjugate more readily internalized by a cell, and more readily subject to active control of release rate inside the cell.

[0012]Accordingly, in one aspect, the present invention features biologically active peptides conjugated to chemical compounds through a carbamate linkage. Conjugates of the present invention provide numerous advantages, such as retention of the biological activity of the peptide, enhanced stability of the conjugate in plasma, and intracellular release of the attached compound. Preferred peptides for use with conjugates of the invention include somatostatin, somatostatin analogs, bombesin, bombesin analogs, KiSS peptides and analogs, urotensin II peptides and analogs, GnRH I and II peptides and analogs, octreotide, depreotide, vapreotide, vasoactive intestinal peptide (VIP), cholecystokinin (CCK), insulin-like growth factor (IGF), RGD-containing peptides, melanocyte-stimulating hormone (MSH) peptide, neurotensin, calcitonin, peptides from complementarity determining regions of an antitumor antibody, glutathione; YIGSR (leukocyte-avid peptides, e.g., P483H, which contain the heparin-binding region of platelet factor-4 (PF-4) and a lysine-rich sequence), atrial natriuretic peptide (ANP), β-amyloid peptides, delta-opioid antagonists (e.g., [125I]ITIPP(psi) [H-Tyr(3′I)-Ticpsi[CH2NH]Phe-Phe-OH]; ITIPP(psi)), annexin-V, endothelin, interleuking (IL)-1, IL-1ra, IL-2, and IL-8, leukotriene B4 (LTB4), chemotactic peptides (e.g., N-formyl-methionyl-leucyl-phenylalanine-lysine (fMLFK)), GP IIb / IIIa receptor antagonists (e.g., DMP444), epidermal growth factor, human neutrophil elastase inhibitor (e.g., EPI-HNE-2 and EPI-HNE-4), plasmin inhibitor, antimocrobial peptides, apticide (e.g., P280 and P274), thrombospondin receptor (including analogs such as TP-1300), bitistatin, pituitary adenylyl cyclase type I receptor (PAC1), fibrin α-chain, and derivatives and analogs thereof. Also included are peptides derived from a phage display library, and conservative substitutions thereof, that are targeted to a cell or tissue in the body of a mammal (e.g., diseased tissue, such as a tumor or a proliferative angiogenic blood vessel; see, e.g., Aina et al., Biopolymers 66:184-199, 2002). Such peptides are useful for specifically targeting therapeutic agents and cytotoxic agents to a cell, such as a cancer cell, or to a tissue in the body of a mammal (e.g., the delivery of anti-apoptotic drugs to cardiac or brain tissue), or to selectively target white blood cells or tubercles infected with tuberculosis. For example, when somatostatin or bombesin is used as the biologically active peptide in a conjugate of the invention, the therapeutic or cytotoxic agent may be targeted to a cancer cell that expresses a somatostatin receptor or a bombesin receptor. The invention also features antibodies (e.g., a monoclonal antibody) or a fragment thereof, that can be linked to the conjugate of the invention. As discussed above with respect to peptides, when an antibody is incorporated in a conjugate of the invention, the therapeutic or cytotoxic agent may be targeted to specific antibody binding sites.

Problems solved by technology

Commonly used ester derivatives are a particularly poor choice of linker because peripherally circulating compound is rapidly attacked by ubiquitous esterases.

However, this linker strategy is not optimal for peptide conjugates because most of the work-up and purification of synthetic peptides is done in acidic media.

The toxic side effects of many of these therapies, as well as standard treatments of neoplastic disease, effectively limit the amount of active agent that may be administered to a patient.

Additionally, many active agents cause organ-specific toxicities, further limiting the dose that may be delivered to the target tissue.

For instance, the cardiotoxicity of many anthracycline family members reduces the maximum therapeutic dose available for this group of chemotherapeutic agents.

The physical and chemical properties of many cytotoxic agents make drug conjugation to biologically active peptides, such as somatostatin and bombesin, problematic.

For example, the drug may reduce the specificity of binding or the biological activity of the peptide analog, limiting its effectiveness as a targeting agent.

Images