Compounds and methods for inhibiting cancer metastasis

Abstract

Agents for inhibiting cancer metastasis are provided. The methods comprise administering to a patient an antimetastatic agent that comprises one or more of: (a) a peptide sequence that is at least 50% identical to an OB-cadherin CAR sequence; (b) a non-peptide mimetic of an OB-cadherin CAR sequence; (c) a substance, such as an antibody or antigen-binding fragment thereof, that specifically binds an OB-cadherin CAR sequence; and/or (d) a polynucleotide encoding a polypeptide that comprises an OB-cadherin CAR sequence or analogue thereof

What is claimed is:  
     
         1 . A method for inhibiting metastasis of a cancer in a patient, comprising administering to a patient an antimetastatic agent that comprises an OB-cadherin CAR sequence, or an analogue thereof that is at least 50% identical to an OB-cadherin CAR sequence such that the ability to inhibit OB-cadherin mediated cell adhesion is not substantially diminished relative to an OB-cadherin CAR sequence.  
     
     
         2 . A method according to  claim 1 , wherein the agent is a peptide consisting of 3-50 amino acid residues linked by peptide bonds.  
     
     
         3 . A method according to  claim 1 , wherein the agent is a peptide consisting of 4-16 amino acid residues linked by peptide bonds.  
     
     
         4 . A method according to  claim 2 , wherein the agent comprises one or more OB-cadherin CAR sequences selected from the group consisting DDK, IDDK (SEQ ID NO:37) DDKS (SEQ ID NO:38), VIDDK (SEQ ID NO:39), IDDKS (SEQ ID NO:40), VIDDKS (SEQ ID NO:41), DDKSG (SEQ ID NO:42), IDDKSG (SEQ ID NO:43), VIDDKSG (SEQ ID NO:44), FVIDDK (SEQ ID NO:45), FVIDDKS (SEQ ID NO:46), FVIDDKSG (SEQ ID NO:47), IFVIDDK (SEQ ID NO:48), IFVIDDKS (SEQ ID NO:49), IFVIDDKSG (SEQ ID NO:50), EEY, IEEY (SEQ ID NO:51), EEYT (SEQ ID NO:52), VIEEY (SEQ ID NO:53), IEEYT (SEQ ID NO:54), VIEEYT (SEQ ID NO:55), EEYTG (SEQ ID NO:56), IEEYTG (SEQ ID NO:56), VIEEYTG (SEQ ID NO:58), FVIEEY (SEQ ID NO:59), FVIEEYT (SEQ ID NO:60), FVIEEYTG (SEQ ID NO:61), FFVIEEY (SEQ ID NO:62), FFVIEEYT (SEQ ID NO:63), FFVIEEYTG (SEQ ID NO:64), EAQ, VEAQ (SEQ ID NO:65), EAQT (SEQ ID NO:66), SVEAQ (SEQ ID NO:67), VEAQT (SEQ ID NO:68), SVEAQT (SEQ ID NO:69), EAQTG (SEQ ID NO:70), VEAQTG (SEQ ID NO:71), SVEAQTG (SEQ ID NO:72), FSVEAQ (SEQ ID NO:73), FSVEAQT (SEQ ID NO:74), FSVEAQTG (SEQ ID NO:75), YFSVEAQ (SEQ ID NO:76), YFSVEAQT (SEQ ID NO:77) and YFSVEAQTG (SEQ ID NO:78).  
     
     
         5 . A method according to  claim 4 , wherein the agent comprises a linear peptide having the sequence N—Ac—IFVIDDKSG—NH 2  (SEQ ID NO:50), N—Ac—FFVIEEYTG—NH 2  (SEQ ID NO:64) or N—Ac—YFSVEAQTG—NH 2  (SEQ ID NO:78).  
     
     
         6 . A method according to  claim 1 , wherein the CAR sequence is present within a cyclic peptide.  
     
     
         7 . A method according to  claim 6 , wherein the cyclic peptide has the formula:  
       
         
           
           
               
               
           
         
         wherein W is a tripeptide selected from the group consisting of EEY, DDK and EAQ;  
         wherein X 1 , and X 2  are optional, and if present, are independently selected from the group consisting of amino acid residues and combinations thereof in which the residues are linked by peptide bonds, and wherein X 1  and X 2  independently range in size from 0 to 10 residues, such that the sum of residues contained within X 1  and X 2  ranges from 1 to 12;  
         wherein Y 1  and Y 2  are independently selected from the group consisting of amino acid residues, and wherein a covalent bond is formed between residues Y 1  and Y 2 ; and  
         wherein Z 1  and Z 2  are optional, and if present, are independently selected from the group consisting of amino acid residues and combinations thereof in which the residues are linked by peptide bonds.  
       
     
     
         8 . A method according to  claim 7 , wherein the agent has a sequence selected from the group consisting of:  CDDKC  (SEQ ID NO:79),  CIDDKC  (SEQ ID NO:80),  CDDKSC  (SEQ ID NO:81),  CVIDDKC  (SEQ ID NO:82),  CIDDKSC  (SEQ ID NO:83),  CVIDDKSC  (SEQ ID NO:84),  CDDKSGC  (SEQ ID NO:85),  CIDDKSGC  (SEQ ID NO:86),  CVIDDKSGC  (SEQ ID NO:87),  CFVIDDKC  (SEQ ID NO:88),  CFVIDDKSC  (SEQ ID NO:89),  CFVIDDKSGC  (SEQ ID NO:90),  CIFVIDDKC  (SEQ ID NO:91),  CIFVIDDKSC  (SEQ ID NO:92),  CIFVIDDKSGC  (SEQ ID NO:93),  DDDKK  (SEQ ID NO:94),  DIDDKK  (SEQ ID NO:95),  DVIDDKK  (SEQ ID NO:96),  DFVIDDKK  (SEQ ID NO97),  DIFVfDDKK  (SEQ ID NO:98),  EDDKK  (SEQ ID NO:99),  EIDDKK  (SEQ ID NO:100),  EVIDDKK  (SEQ ID NO:101),  EFVIDDKK  (SEQ ID NO:102),  EIFVIDDKK  (SEQ ID NO:103),  FVIDDK  (SEQ ID NO:104),  FVIDDKS  (SEQ ID NO:105),  FVIDDKSG  (SEQ ID NO:106),  KDDKD  (SEQ ID NO:107),  KIDDKD  (SEQ ID NO:108),  KDDKSD  (SEQ ID NO:109),  KVIDDKD  (SEQ ID NO:110),  KIDDKSD  (SEQ ID NO:111),  KVIDDKSD  (SEQ ID NO:112),  KDDKSGD  (SEQ ID NO:113),  KIDDKSGD  (SEQ ID NO:114),  KVIDDKSGD  (SEQ ID NO:115),  KFVIDDKD  (SEQ ID NO:116),  KFVIDDKSD  (SEQ ID NO:117),  KFVIDDKSGD  (SEQ ID NO:118),  KIFVIDDKD  (SEQ ID NO: 119),  KIFVDDKSD  (SEQ ID NO:120),  KIFVIDDKSGD  (SEQ ID NO:121),  VIDDK  (SEQ ID NO:122),  IDDKS  (SEQ ID NO:123),  VIDDKS  (SEQ ID NO:124),  VIDDKSG  (SEQ ID NO:125),  DDKSG  (SEQ ID NO:126),  IDDKSG  (SEQ ID NO:127),  IFVIDDK  (SEQ ID NO:128),  IFVDDKS  (SEQ ID NO:129),  IFVIDDKSG  (SEQ ID NO:130),  KDDKE  (SEQ ID NO:131),  KIDDKE  (SEQ ID NO:132),  KDDKSE  (SEQ ID NO:133),  KVIDDKE  (SEQ ID NO:134),  KIDDKSE  (SEQ ID NO:135),  KVIDDKSE  (SEQ ID NO:136),  KDDKSGE  (SEQ ID NO:137),  KIDDKSGE  (SEQ ID NO:138),  KVIDDKSGE  (SEQ ID NO:139),  KFVIDDKE  (SEQ ID NO:140),  KFVIDDKSE  (SEQ ID NO:141),  KFVIDDKSGE  (SEQ ID NO:142),  KIFVIDDKE  (SEQ ID NO:143),  KIFVVDDKSE  (SEQ ID NO:144),  KIFVIDDKSGE  (SEQ ID NO:145),  CEEYC  (SEQ ID NO:146),  CIEEYC  (SEQ ID NO:147),  CEEYTC  (SEQ ID NO:148),  CVIEEYC  (SEQ ID NO:149),  CIEEYTC  (SEQ ID NO:150),  CVIEEYTC  (SEQ ID NO:151),  CEEYTGC  (SEQ ID NO:152),  CIEEYTGC  (SEQ ID NO:153),  CVIEEYTGC  (SEQ ID NO:154),  CFVIEEYC  (SEQ ID NO:155),  CFVIEEYTC  (SEQ ID NO 156),  CFVIEEYTGC  (SEQ ID NO:157),  CFFVIEEYC  (SEQ ID NO:158),  CFFVIEEYTC  (SEQ ID NO:159),  CFFVIEEYTGC  (SEQ ID NO:160),  KEEYD   0  (SEQ ID NO:161),  KIEEYD  (SEQ ID NO:162),  KEEYTD  (SEQ ID NO:163),  KVIEEYD  (SEQ ID NO:164),  KIEEEYTD  (SEQ ID NO:165),  KVIEEYTD  (SEQ ID NO:166),  KEEYTGCD  (SEQ ID NO:167),  KIEEYTGD  (SEQ ID NO:168),  KVIEEYTGD  (SEQ ID NO:169),  KFVIEEYD  (SEQ ID NO:170),  KFVIEEYTD  (SEQ ID NO:171),  KFVIEEYTGD  (SEQ ID NO:172),  KFFVIEEYD  (SEQ ID NO:173),  KFFVIEEYTD  (SEQ ID NO:174),  KFFVIEEYTGD  (SEQ ID NO:175),  EEEYK  (SEQ ID NO:176),  EIEEYK  (SEQ ID NO:177),  EEEYTK  (SEQ ID NO:178),  EVIEEYK  (SEQ ID NO:179),  EIEEYTK  (SEQ ID NO: 180),  EVIEEYTK  (SEQ ID NO: 181),  EEEYTGK  (SEQ ID NO: 182),  EIEEYTGK  (SEQ ID NO:183),  EVIEEYTGK  (SEQ ID NO:184),  EFVIEEYK  (SEQ ID NO:185),  EFVIEEYTK  (SEQ ID NO: 186),  EFVIEEYTGK  (SEQ ID NO: 187),  EFFVIEEYK  (SEQ ID NO:188),  EFFVIEEYTK  (SEQ ID NO:189),  EFFVIEEYTGK  (SEQ ID NO:190),  DCEEYK  (SEQ ID NO:191),  DIEEYCK  (SEQ ID NO:192),  DEEYTK  (SEQ ID NO:193),  DVIEEYK  (SEQ ID NO:194),  DIEEYTK  (SEQ ID NO:195),  DVIEEYTK  (SEQ ID NO:196),  DEEYTGK  (SEQ ID NO:197),  DIEEYTGK  (SEQ ID NO:198),  DVIEEYTGK  (SEQ ID NO:199),  DFVIEEYK  (SEQ ID NO:200),  DFVIEEYTK  (SEQ ID NO:201),  DFVIEEYTGK  (SEQ ID NO:202),  DFFVIEEYK  (SEQ ID NO:7203),  DFFVIEEYTK  (SEQ ID NO:204),  DFFVIEEYTGK  (SEQ ID NO:205),  KEEYE  (SEQ ID NO:206),  KIEEYE  (SEQ ID NO:207),  KEEYTE  (SEQ ID NO:208),  KVIEEYE  (SEQ ID NO:209),  KIEEYTE  (SEQ ID NO:210),  KVIEEYTE  (SEQ ID NO:211),  KEEYTGE  (SEQ ID NO:212),  KIEEYTGE  (SEQ ID NO:213),  KVIEEYTGE  (SEQ ID NO:214),  KFVIEEYE  (SEQ ID NO:215),  KFVIEEYTE  (SEQ ID NO:216),  KFVIEEYTGE  (SEQ ID NO:217),  KFFVIEEYE  (SEQ ID NO:218),  KFFVIEEYTE  (SEQ ID NO:219),  KFFVIEEYTGE  (SEQ ID NO:220),  VIEEY  (SEQ ID NO:221),  IEEYT  (SEQ ID NO:222),  VIEEYT  (SEQ ID NO:223),  EEYTG  (SEQ ID NO:224),  IEEYTG  (SEQ ID NO:225),  VIEEYTG  (SEQ ID NO:226),  FVIEEY (SEQ ID NO:227),  FVIEEYT  (SEQ ID NO:228),  FVIEEYTG  (SEQ ID NO:229),  FFVIEEY  (SEQ ID NO:230),  FFVIEEYT  (SEQ ID NO:231),  FFVIEEYTG  (SEQ ID NO:232),  CEAQC  (SEQ ID NO:233),  CVEAQC  (SEQ ID NO:234),  CEAQTC  (SEQ ID NO:235),  CSVEAQC  (SEQ ID NO:236),  CVEAQTC  (SEQ ID NO:237),  CSVEAQTC  (SEQ ID NO:238),  CEAQTGC  (SEQ ID NO:239),  CVEAQTGC  (SEQ ID NO:240),  CSVEAQTGC  (SEQ ID NO:241),  CFSVEAQC  (SEQ ID NO:242),  CFSVEAQTC  (SEQ ID NO:243),  CFSVEAQTGC  (SEQ ID NO:244),  CYFSVEAQC  (SEQ ID NO:245),  CYFSVEAQTC  (SEQ ID NO:246),  CYFSVEAQTGC  (SEQ ID NO:247),  KEAQD  (SEQ ID NO:248),  KVEAQD  (SEQ ID NO:249),  KEAQTD  (SEQ ID NO:250),  KSVEAQD  (SEQ ID NO:251),  KVEAQTD  (SEQ ID NO:252),  KSVEAQTD  (SEQ ID NO:253),  KEAQTGD  (SEQ ID NO:254),  KVEAQTGD  (SEQ ID NO:255),  KSVEAQTGD  (SEQ ID NO:256),  KFSVEAQD  (SEQ ID NO:257),  KFSVEAQTD  (SEQ ID NO:258),  KFSVEAQTGD  (SEQ ID NO:259),  KYFSVEAQD  (SEQ ID NO:260),  KYFSVEAQTD  (SEQ ID NO:261),  KYFSVEAQTGD  (SEQ ID NO:262),  EEAQK  (SEQ ID NO:263),  EVEAQK  (SEQ ID NO:264),  EEAQTK  (SEQ ID NO:265),  ESVEAQK  (SEQ ID NO:266),  EVEAQTK  (SEQ ID NO:267),  ESVEAQTK  (SEQ ID NO:268),  EEAQTGK  (SEQ ID NO:269),  EVEAQTGK  (SEQ ID NO:270),  ESVEAQTGK  (SEQ ID NO:271),  EFSVEAQK  (SEQ ID NO:272),  EFSVEAQTK  (SEQ ID NO:273),  EFSVEAQTGK  (SEQ ID NO:274),  EYFSVEAQK  (SEQ ID NO:275),  EYFSVEAQTK  (SEQ ID NO:276),  EYFSVEAQTGK  (SEQ ID NO:277),  DEAQK  (SEQ ID NO:278),  DVEAQK  (SEQ ID NO:279),  DEAQTK  (SEQ ID NO:280),  DSVEAQK  (SEQ ID NO:281),  DVEAQTK  (SEQ ID NO:282),  DSVEAQTK  (SEQ ID NO:283),  DEAQTGK  (SEQ ID NO:284),  DVEAQTGK  (SEQ ID NO:285),  DSYEAQTGK  (SEQ ID NO:286),  DFSVEAQK  (SEQ ID NO:287),  DFSVEAQTK  (SEQ ID NO:288),  DFSVEAQTGK  (SEQ ID NO:289),  DYFSVEAQK  (SEQ ID NO:290),  DYFSVEAQTK  (SEQ ID NO:291),  DYFSVEAQTGK  (SEQ ID NO:292),  KEAQE  (SEQ ID NO:293),  KVEAQE  (SEQ ID NO:294),  KEAQTE  (SEQ ID NO:295),  KSVEAQE  (SEQ ID NO:296),  KVEAQTE  (SEQ ID NO:297),  KSVEAQTE  (SEQ ID NO:298),  KEAQTGE  (SEQ ID NO:299),  KVEAQTGE  (SEQ ID NO:300),  KSVEAQTGE  (SEQ ID NO:301),  KFSVEAQE  (SEQ ID NO:302),  KFSVEAQTE  (SEQ ID NO:303),  KFSVEAQTGE  (SEQ ID NO:304),  KYFSVEAQE  (SEQ ID NO:305),  KYFSVEAQTE  (SEQ ID NO:306),  KYFSVEAQTGE  (SEQ ID NO:307),  SVEAQ  (SEQ ID NO:308),  VEAQT  (SEQ ID NO:309),  SVEAQT  (SEQ ID NO:310),  EAQTG  (SEQ ID NO:311),  VEAQTG  (SEQ ID NO:312),  SVEAQTG  (SEQ ID NO:313),  FSVEAQ  (SEQ ID NO:314),  FSVEAQT  (SEQ ID NO:315),  FSVEAQTG  (SEQ ID NO:316),  YFSVEAQ  (SEQ ID NO:317),  YFSVEAQT  (SEQ ID NO:318) and  YFSVEAQTG  (SEQ ID NO:319).  
     
     
         9 . A method according to  claim 1 , wherein the agent is linked to a drug.  
     
     
         10 . A method according to  claim 1 , wherein the agent is linked to a targeting moiety.  
     
     
         11 . A method according to  claim 1 , wherein the agent is linked to: 
 (a) a CAR sequence that is specifically recognized by an adhesion molecule other than OB-cadherin; and/or    (b) an antibody or antigen-binding fragment thereof that specifically binds to a CAR sequence that is specifically recognized by an adhesion molecule other than OB-cadherin.    
     
     
         12 . A method according to  claim 10 , wherein the adhesion molecule is selected from the group consisting of cadherins, integrins, occludin, claudins and members of the immunoglobulin superfamily.  
     
     
         13 . A method according to  claim 1 , wherein the agent is present within a pharmaceutical composition comprising a physiologically acceptable carrier.  
     
     
         14 . A method according to  claim 12 , wherein the composition further comprises a drug.  
     
     
         15 . A method according to  claim 12 , wherein the composition further comprises a modulator of cell adhesion comprising: 
 (a) a CAR sequence that is specifically recognized by an adhesion molecule other than OB-cadherin; and/or    (b) an antibody or antigen-binding fragment thereof that specifically binds to a CAR sequence that is specifically recognized by an adhesion molecule other than OB-cadherin.    
     
     
         16 . A method according to  claim 14 , wherein the adhesion molecule is selected from the group consisting of cadherins, integrins, occludin, claudins and members of the immunoglobulin superfamily.  
     
     
         17 . A method for inhibiting metastasis of a cancer in a patient, comprising administering to a patient a polynucleotide encoding an antimetastatic agent that comprises an, OB-cadherin CAR sequence, or an analogue thereof that is at least 50% identical to an OB-cadherin CAR sequence such that the ability to inhibit OB-cadherin mediated cell adhesion is not substantially diminished relative to an OB-cadherin CAR sequence.  
     
     
         18 . A method according to  claim 16 , wherein the agent is a peptide consisting of 3-50 amino acid residues linked by peptide bonds.  
     
     
         19 . A method for inhibiting metastasis of a cancer in a patient, comprising administering to a patient an antimetastatic agent comprising an antibody or antigen-binding fragment thereof that specifically binds to an OB-cadherin CAR sequence.  
     
     
         20 . A method according to  claim 19 , wherein the OB-cadherin CAR sequence is selected from the group consisting of IFVIDDKSG (SEQ ID NO:50), FFVIEEYTG (SEQ ID NO:64) and YFSVEAQTG (SEQ ID NO:78).  
     
     
         21 . A method according to  claim 19 , wherein the agent is linked to a drug.  
     
     
         22 . A method according to  claim 19 , wherein the agent is linked to a targeting moiety.  
     
     
         23 . A method according to  claim 19 , wherein the agent is linked to: 
 (a) a CAR sequence that is specifically recognized by an adhesion molecule other than OB-cadherin; and/or    (b) an antibody or antigen-binding fragment thereof that specifically binds to a CAR sequence that is specifically recognized by an adhesion molecule other than OB-cadherin.    
     
     
         24 . A method according to  claim 23 , wherein the adhesion molecule is selected from the group consisting of cadherins, integrins, occludin, claudins and members of the immunoglobulin superfamily.  
     
     
         25 . A method according to  claim 19 , wherein the agent is present within a pharmaceutical composition comprising a physiologically acceptable carrier.  
     
     
         26 . A method according to  claim 25 , wherein the composition further comprises a drug.  
     
     
         27 . A method according to  claim 25 , wherein the composition further comprises a modulator of cell adhesion comprising: 
 (a) a CAR sequence that is specifically recognized by an adhesion molecule other than OB-cadherin; and/or    (b) an antibody or antigen-binding fragment thereof that specifically binds to a CAR sequence that is specifically recognized by an adhesion molecule other than OB-cadherin.    
     
     
         28 . A method according to  claim 27 , wherein the adhesion molecule is selected from the group consisting of cadherins, integrins, occludin, claudins and members of the immunoglobulin superfamily.  
     
     
         29 . A method for removing metastatic cells from a biological sample, comprising the steps of: 
 (a) contacting a biological sample with an antimetastatic agent that comprises: 
 (i) an OB-cadherin CAR sequence or an analogue thereof that is at least 50% identical to an OB-cadherin CAR sequence such that the ability to inhibit OB-cadherin mediated cell adhesion is not substantially diminished relative to an OB-cadherin CAR sequence; or  
 (ii) an antibody or antigen-binding fragment thereof that specifically binds to an OB-cadherin CAR sequence; 
 wherein the contact takes place under conditions and for a time sufficient to allow OB-cadherin-expressing cells within the biological sample to bind to the antimetastatic agent; and  
 
   (b) separating OB-cadherin expressing cells bound to the antimetastatic agent from the biological sample.    
     
     
         30 . A method according to  claim 29 , wherein the antimetastatic agent is linked to a support material.  
     
     
         31 . A method according to  claim 29 , wherein the biological sample is further contacted with a modulator of cell adhesion comprising: 
 (a) a CAR sequence that is specifically recognized by an adhesion molecule other than OB-cadherin; and/or    (b) an antibody or antigen-binding fragment thereof that specifically binds to a CAR sequence that is specifically recognized by an adhesion molecule other than OB-cadherin.    
     
     
         32 . A method according to  claim 31 , wherein the adhesion molecule is selected from the group consisting of cadherins, integrins, occludin, claudins and members of the immunoglobulin superfamily.  
     
     
         33 . A method according to  claim 29 , wherein the biological sample is blood or bone marrow obtained from a patient.  
     
     
         34 . A method for removing metastatic cells from a patient, comprising the steps of: 
 (a) obtaining a biological sample from a patient;    (b) contacting the biological sample with an antimetastatic agent that comprises: 
 (i) an OB-cadherin CAR sequence or an analogue thereof that is at least 50% identical to an OB-cadherin CAR sequence such that the ability to inhibit OB-cadherin mediated cell adhesion is not substantially diminished relative to an OB-cadherin CAR sequence; or  
 (ii) an antibody or antigen-binding fragment thereof that specifically binds to an OB-cadherin CAR sequence; 
 wherein the contact takes place under conditions and for a time sufficient to allow OB-cadherin-expressing cells within the biological sample to bind to the antimetastatic agent; and  
 
   (c) separating OB-cadherin expressing cells bound to the antimetastatic agent from the remainder of the biological sample; and    (d) returning the remainder of the biological sample to the patient.    
     
     
         35 . A method according to  claim 34 , wherein the antimetastatic agent is linked to a support material.  
     
     
         36 . A method according to  claim 34 , wherein the biological sample is further contacted with a modulator of cell adhesion comprising: 
 (a) a CAR sequence that is specifically recognized by an adhesion molecule other than OB-cadherin; and/or    (b) an antibody or antigen-binding fragment thereof that specifically binds to a CAR sequence that is specifically recognized by an adhesion molecule other than OB-cadherin.    
     
     
         37 . A method according to  claim 36 , wherein the adhesion molecule is selected from the group consisting of cadherins, integrins, occludin, claudins and members of the immunoglobulin superfamily.  
     
     
         38 . A method according to  claim 34 , wherein the biological sample is blood or bone marrow.