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.

1 - 114 . (canceled) 
     
     
         115 . A compound represented by the following formula: 
       
         
           
           
               
               
           
         
         wherein: 
         X is a cytotoxic agent or therapeutic agent; 
         n is an integer from 0 to 6, wherein (CH 2 ) n  is an alkyl or a cyclic group; 
         R is N(R 1 R 2 ), OR 1 , or SR 1 , wherein R 1  and R 2  are, independently, hydrogen or a straight or branched chain lower alkyl group having fewer than 11 carbon atoms, and wherein R 3  is a NH(CH 2 ) m SH group and m=2 to 6, D or L cysteine, a benzophenone, an OH group, or is NH—Y-Z-Q 
         wherein: 
         Y is a hydrophilic peptide, a hydrophilic polymer, or is omitted; 
         Z is a linking peptide that is bonded to Q at the N-terminus or at a compatible side-chain amino group of Q and has the formula: 
         A-B-C-E-F, wherein:
 A is D-Lys, D-Tyr, D-Ser, or L-Ser, or is deleted; 
 B is D-Lys or D-Tyr, or is deleted; 
 C is Lys, Ser, hSer, Thr, Nle, Abu, Nva, 2-pyridyl-Ala, 3-pyridyl-Ala, 4-pyridyl-Ala, Orn, Dab, Dap, 4-NH 2 -Phe, D-4-OH-Pro, or L-4-OH-Pro, or is deleted; 
 E is D-Lys, D-Tyr, D-Ser, D-4-OH-Pro, L-4-OH-Pro, 3-iodo-D-Tyr, 3-5 diiodo-D-Tyr, 3-astatine-D-Tyr, 3-5 astatine-D-Tyr, 3-bromo-D-Tyr, 3-5 dibromo-D-Tyr, D-Asn, L-Asn, D-Asp, L-Asp, D-Glu, L-Glu, D-Gln, or L-Gln; and 
 F is D-Lys, D-Tyr, D-Ser, L-Ser, D-4-OH-Pro, L-4-OH-Pro, 3-iodo-D-Tyr, 3-5 diiodo-D-Tyr, 3-astatine-D-Tyr, 3-5 astatine-D-Tyr, 3-bromo-D-Tyr, 3-5 dibromo-D-Tyr, D-Asn, L-Asn, D-Asp, L-Asp, D-Glu, L-Glu, D-Gln, or L-Gln; 
 
         provided that when A, B, C, and E are Tyr, Tyr, Lys, and Tyr, respectively, F is not Lys; and when A, B, C, and E are Lys, Tyr, Lys, and Tyr, respectively, F is not Tyr or Lys; and when A and B are deleted, and C and E are Lys and Tyr, respectively, F is not Tyr or Lys; and 
         Q is a targeting moiety. 
       
     
     
         116 . The compound of  claim 115 , wherein R 3  is NH—Y-Z-Q and the compound is a conjugate compound represented by the following formula: 
       
         
           
           
               
               
           
         
       
     
     
         117 . The compound of  claim 115 , wherein X is a cytotoxic agent. 
     
     
         118 . The compound of  claim 117 , wherein said cytotoxic agent is an alkylating agent, an antibiotic, an antimetabolite, a tubulin inhibitor, a topoisomerase I or II inhibitor, an hormonal agonist or antagonists, an apoptotic agent, or an immunomodulator, or is camptothecin, homocamptothecin, colchicine, thiocolchicine, combretastatin, dolistatin, doxorubicin, methotrexate, podophyllotoxin, rhizoxin, rhizoxin D, a taxol, paclitaxel, CC1065, or a maytansinoid. 
     
     
         119 . The compound of  claim 115 , wherein Y is said hydrophilic peptide. 
     
     
         120 . The compound of  claim 119 , wherein at least one V is a D-amino acid. 
     
     
         121 . The compound of  claim 119 , wherein V is independently D-Ser or L-Ser. 
     
     
         122 . The compound of  claim 115 , wherein Y is said hydrophilic polymer. 
     
     
         123 . The compound of  claim 115 , wherein X is somatostatin, bombesin, a KiSS peptide, a urotensin II peptide, gonadotropin-releasing hormone (GnRH) I and II peptides, octreotide, depreotide, vapreotide, vasoactive intestinal peptide (VIP), cholecystokinin (CCK), insulin-like growth factor (IGF), RGD-containing peptides, melanocyte-stimulating hormone (MSH) peptide, neurotensin, calcitonin, a peptide comprising the complementarity determining region of an antitumor antibody, glutathione, a leukocyte-avid peptide comprising the amino acid sequence YIGSR, the heparin-binding region of platelet factor-4 (PF-4), and a lysine-rich sequence, atrial natriuretic peptide (ANP), a β-amyloid peptide, a delta-opioid antagonist, annexin-V, endothelin, interleukin (IL)-1, IL-1ra, IL-2, IL-8, leukotriene B4 (LTB4), a chemotactic peptide, a GP IIb/IIIa receptor antagonist, epidermal growth factor, a human neutrophil elastase inhibitor, plasmin inhibitor, an antimicrobial peptide, apticide P280, apticide P274, a thrombospondin receptor, bitistatin, pituitary adenylyl cyclase type I receptor (PAC1), fibrin α-chain, or derivatives or analogs thereof. 
     
     
         124 . The compound of  claim 123 , wherein Q is said somatostatin peptide. 
     
     
         125 . The compound of  claim 123 , wherein Q is said bombesin peptide. 
     
     
         126 . The compound of  claim 115 , wherein Z is D-Ser-Nle-D-Ser-D-Ser, D-Ser-Lys-D-Ser-D-Ser, D-Ser-Lys-D-Tyr-D-Tyr, D-Ser-Lys-D-Tyr-D-Ser, D-Ser-Ser-D-Lys-D-Ser, D-Ser-Ser-D-Lys-Ser, D-Ser-Nle-D-Tyr-D-Ser, D-Ser-Pal-D-Tyr-D-Ser, D-Ser-Thr-D-Tyr-D-Ser, Lys-D-Ser-D-Ser, Ser-D-Lys-D-Ser, Ser-D-Lys-Ser, Nle-D-Tyr-D-Ser, Lys-D-Tyr-D-Ser, Pal-D-Lys-D-Ser, Thr-D-Tyr-D-Ser, D-Ser-D-Lys, D-Ser-D-Tyr, D-Lys-D-Lys, D-Lys-D-Tyr, or D-Tyr-D-Lys, or wherein Z has the formula:
 E-F, wherein:
 E is D-Lys, D-Tyr, D-Ser, D-4-OH-Pro, L-4-OH-Pro, 3-iodo-D-Tyr, 3-5 diiodo-D-Tyr, 3-astatine-D-Tyr, 3-5 astatine-D-Tyr, 3-bromo-D-Tyr, 3-5 dibromo-D-Tyr, D-Asn, L-Asn, D-Asp, L-Asp, D-Gln, or L-Gln; and 
 F is D-Lys, D-Tyr, D-Ser, L-Ser, D-4-OH-Pro, L-4-OH-Pro, 3-iodo-D-Tyr, 3-5 diiodo-D-Tyr, 3-astatine-D-Tyr, 3-5 astatine-D-Tyr, 3-bromo-D-Tyr, 3-5 dibromo-D-Tyr, D-Asn, L-Asn, D-Asp, L-Asp, D-Glu, L-Glu, D-Gln, or L-Gln. 
   
     
     
         127 . The compound of  claim 115 , wherein R comprises 1 to 8 carbon atoms. 
     
     
         128 . The compound of  claim 115 , wherein the R 3  group is used to attach a peptide, protein, or antibody to said compound. 
     
     
         129 . The compound of  claim 128 , wherein R 3  is NH(CH 2 )mSH and m=2 to 6, and wherein said peptide, protein, or antibody is attached to said compound by a thiol reaction. 
     
     
         130 . The compound of  claim 128 , wherein R 3  is a benzophenone and said peptide, protein, or antibody is attached to said compound by a photochemical reaction. 
     
     
         131 . The compound of  claim 130 , wherein said benzophenone is p-benzoyl-phenylalanine. 
     
     
         132 . The compound of  claim 115 , wherein n=2 or 3. 
     
     
         133 . The compound of  claim 116  admixed with a pharmaceutically acceptable carrier, excipient, or salt. 
     
     
         134 . The compound of  claim 124 , wherein said somatostatin peptide is cyclo[Cys-Phe-D-Trp-Lys-Thr-Cys]-Thr-NH 2 . 
     
     
         135 . The compound of  claim 134 , wherein said compound comprises camptothecin as X, D-Ser-Nle-D-Tyr-D-Ser as Z, N(R 1 R 2 ) as R, wherein R 1  and R 2  are hydrogen and methyl, respectively, and wherein Y is omitted. 
     
     
         136 . A method of treating a disease comprising administering to a subject suffering from said disease a therapeutically effective amount of the compound of  claim 116 , wherein the disease is inflammatory bowel disease, rheumatoid arthritis, acromegaly, tuberculosis, a tumor of the lung, breast, brain, eye, prostate or colon, a tumor of neuroendocrine origin, an angiogenesis disease that causes inappropriate proliferation of blood vessels, retinal macular degeneration or diabetic neuropathy. 
     
     
         137 . The method of  claim 136 , comprising administering the compound of  claim 135 . 
     
     
         138 . The method of  claim 136 , wherein said tumor of neuroendocrine origin is carcinoid syndrome. 
     
     
         139 . The method of  claim 136 , wherein said blood vessels are in the eye.