US2010144671A1PendingUtilityA1

Compositions and Methods for Cancer Treatment

Assignee: WOMEN AND INFANTS HOSPITAL OFPriority: Mar 4, 2005Filed: Mar 3, 2006Published: Jun 10, 2010
Est. expiryMar 4, 2025(expired)· nominal 20-yr term from priority
A61P 35/02A61K 31/59A61P 35/00
48
PatentIndex Score
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Claims

Abstract

Bromoacetoxycalcidiol (B3CD), which is structurally related to calcidiol, exhibits cytotoxic and apoptotic activity toward cancer cells, including highly aggressive neuroblastoma cells. A series of small molecules designed around the structure of B3CD is expected to have growth inhibitory and apoptogenic activities toward a wide range of malignancies. B3CD shows no apparent toxicity in vivo, indicating potential value as a chemotherapeutic agent which will be particularly useful in treating highly aggressive tumors.

Claims

exact text as granted — not AI-modified
1 . A method for inhibiting cancer cell proliferation, comprising administering to said cell an amount of a calcidiol derivative effective to inhibit proliferation of the cancer cell. 
   
   
       2 . The method of  claim 1  wherein the cancer cell is selected from the group consisting of colon, breast, lung, prostate, melanoma, renal, ovarian, bladder, pancreatic, hepatocarcinoma, neuroblastoma, glioblastoma, medulloblastoma, skin, squamous cell carcinoma, acute promyelocytic leukemia, myeloblastic and myelomonocytic cells. 
   
   
       3 . The method of  claim 1  wherein the cancer cell is a neuroblastoma cell. 
   
   
       4 . The method of  claim 3  wherein the neuroblastoma cell is selected from the group consisting of SK-N-SH, SH-SY5Y, SMS-KCN and SMS-KCNR. 
   
   
       5 . The method of  claim 1  wherein the cancer cell is a prostate cancer cell. 
   
   
       6 . The method of  claim 1  wherein the cancer cell is comprised within a mammal. 
   
   
       7 . The method of  claim 6  wherein the mammal is human. 
   
   
       8 . A method of inducing apoptosis in a cancer cell, comprising administering an amount of a calcidiol derivative to a cell effective to induce apoptosis in said cell. 
   
   
       9 . The method of  claim 8 , wherein the calcidiol derivative is bromoacetoxycalcidiol (B3CD). 
   
   
       10 . A method for treating neuroblastoma or prostate cancer, comprising administering to a mammal in need thereof, an effective amount of a calcidiol derivative in a pharmaceutically acceptable vehicle. 
   
   
       11 . The method of  claim 10 , wherein the calcidiol derivative is bromoacetoxycalcidiol (B3CD) in a pharmaceutically acceptable vehicle. 
   
   
       12 . A method for inhibiting TrkB signaling comprising administering to a mammal exhibiting neuroblastoma or prostate cancer an amount of a calcidiol derivative in a pharmaceutically acceptable vehicle effective to suppress BDNF and inhibit angiogenesis. 
   
   
       13 . The method  claim 12 , wherein the calcidiol derivative is bromoacetoxycalcidiol (B3CD) in a pharmaceutically acceptable vehicle effective to suppress BDNF and inhibit angiogenesis. 
   
   
       14 . A method for treating or preventing a disorder associated with uncontrolled cell proliferation, the method comprising administering to a subject in need thereof a composition comprising an effective amount of a calcidiol derivative. 
   
   
       15 . The method of  claim 14 , wherein the calcidiol derivative is bromoacetoxycalcidiol (B3CD). 
   
   
       16 . The method according to  claim 15  wherein the disorder is selected from the group consisting of neuroblastoma, prostate cancer, lymphoma and leukemias. 
   
   
       17 . The method of  claim 15  further comprising administering a second therapeutic agent. 
   
   
       18 . The method of  claim 17  wherein the second therapeutic agent is selected from analgesics, anti-inflammatory agents and antibiotics. 
   
   
       19 . The method of  claim 1 , wherein the calcidiol derivative is a compound of formula I: 
     
       
         
         
             
             
         
       
       wherein: 
       A 1  is single or double bond; 
       A 2  is a single, double or triple bond; 
       X 1  and X 2  are each independently H 2  or CH 2 , provided X 1  and X 2  are not both ═CH 2 ;
 and X 1  can be substituted with CH 2 OR 11    
 wherein R 11  is C(O)alkyl, C(O)aryl, or C(O)aralkyl; 
 
       R 1  is H, OH, or OSi(R 10 ) 3 ;
 wherein R 10  is alkyl, aryl, alkenyl, or aralkyl; 
 
       R 2  is O(CH 2 ) n R 12 ; 
       wherein R 12  is halogen, haloalkyl, amino, alkyl amino, thiol, alkyl thio, hydroxyl, alkoxy, or alkenyl; 
       n is an integer from 1-6; 
       and any of the CH 2  groups may be replaced by CO; 
       R 3  and R 4  independently H, C 1 -C 4  alkyl, hydroxyalkyl, or haloalkyl, or R 3  and R 4  taken together with C 20  form C 3 -C 6  cycloalkyl; 
       R 5  is H, C 1 -C 4  alkyl, hydroxyalkyl, haloalkyl, or carbonyl; 
       R 6  and R 7  are each independently alkyl or haloalkyl;
 R 8  is H or biotin; 
 R 9  is H, hydroxyl, or halogen; and 
 
       pharmaceutically acceptable esters, salts, solvates, clathrates, stereoisomers, enantiomers prodrugs thereof. 
     
   
   
       20 . The method of  claim 19 , wherein the calcidiol derivative is a compound of formula II 
     
       
         
         
             
             
         
       
       wherein X is F, Cl, Br, I, N 3 , N 2 , NH 2 , CN, S—CH 3 , or —N═C═S; 
       R is 
     
     
       
         
         
             
             
         
       
       or a pharmaceutically acceptable salt, solvate, clathrate, stereoismoer, enantiomer or prodrug thereof. 
     
   
   
       21 . The method of  claim 20 , wherein the calcidiol derivative is bromoacetoxycalcidiol (B3CD): 
     
       
         
         
             
             
         
       
       or a pharmaceutically acceptable salt, solvate, clathrate, stereoisomer, enantiomer, or prodrug thereof. 
     
   
   
       22 . The method of  claim 19 , wherein the calcidiol derivative is selected from the group consisting of 
     
       
         
               
               
             
                   
               
                 Bromoacetoxycalcidiol (B3CD) Bromo-acetic acid 3-{2-[1-(5-hydroxy-1,5- dimethyl-hexyl)-7a-methyl-octahydro-inden-4- ylidene]-ethylidene}-4-methylene-cyclohexyl ester 
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
                   
               
                 25-Biotin-Bromoacetoxycalcidiol 
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
                   
               
                 6-(2,4-Dinitro-phenylamino)-hexanoic acid 2- (4-(2-bromo-acetoxy)-2-{2-[1-(5-hydroxy-1,5- dimethyl-hexyl)-7a-methyl-octahydro-inden-4- ylidene]-ethylidene}-cyclohexylidene)-ethyl ester 
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
                   
               
                 Bromo-acetic acid 3-hydroxy-5-{2-[1-(5- hydroxy-1,5-dimethyl-hexyl)-7a-methyl- octahydro-inden-4-ylidene]-ethylidene}-4- methylene-cyclohexyl ester 
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
                   
               
                 Bromo-acetic acid 3-(tert-butyl-dimethyl- silanyloxy)-5-{2-[1-(5-hydroxy-1,5-dimethyl- hexyl)-7a-methyl-octahydro-inden-4-ylidene]- ethylidene}-4-methylene-cyclohexyl ester 
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
                   
               
                 Trifluoro-acetic acid 3-{2-[1-(5-hydroxy-1,5- dimethyl-hexyl)-7a-methyl-octahydro-inden-4- ylidene]-ethylidene}-4-methylene-cyclohexyl ester 
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
                   
               
                 4-Bromo-butyric acid 3-{2-[1-(5-hydroxy-1,5- dimethyl-hexyl)-7a-methyl-octahydro-inden-4- ylidene]-ethylidene}-4-methylene-cyclohexyl ester 
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
                   
               
                 4-Cyano-butyric acid 3-{2-[1-(5-hydroxy-1,5- dimethyl-hexyl)-7a-methyl-octahydro-inden-4- ylidene]-ethylidene}-4-methylene-cyclohexyl ester 
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
                   
               
                 Acrylic acid 3-{2-[1-(5-hydroxy-1,5-dimethyl- hexyl)-7a-methyl-octahydro-inden-4-ylidene]- ethylidene}-4-methylene-cyclohexyl ester 
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
                   
               
                 6-{4-[2-(5-Bromomethoxymethoxy-2- methylene-cyclohexylidene)-ethylidene]-7a- methyl-octahydro-inden-1-yl}-2-methyl-heptan- 2-ol 
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
                   
               
                 6-{4-[2-(5-Aminomethoxymethoxy-2- methylene-cyclohexylidene)-ethylidene]-7a- methyl-octahydro-inden-1-yl}-2-methyl-heptan- 2-ol 
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
                   
               
                 6-{4-[2-(5-Bromomethoxy-2-methylene- cyclohexylidene)-ethylidene]-7a-methyl- octahydro-inden-1-yl}-2-methyl-heptan-2-ol 
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
                   
               
                 2-Methyl-6-{7a-methyl-4-[2-(2-methylene-5- ethylsulfanylmethoxy-cyclohexylidene)- ethylidene]-octahydro-inden-1-yl}-heptan-2-ol 
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
                   
               
                 Bromo-acetic acid 3-(2-{1-[2-(2-hydroxy-2- methyl-propoxy)-1-methyl-ethyl]-7a-methyl- octahydro-inden-4-ylidene}-ethylidene)-4- methylene-cyclohexyl ester 
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
                   
               
                 Bromo-acetic acid 3-{2-[1-(5-hydroxy-1,5- dimethyl-hex-3-ynyl)-7a-methyl-octahydro- inden-4-ylidene]-ethylidene}-4-methylene- cyclohexyl ester 
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
                   
               
                 Bromo-acetic acid 3-{2-[1-(5-hydroxy-1,5- dimethyl-4-oxo-hexyl)-7a-methyl-octahydro- inden-4-ylidene]-ethylidene}-4-methylene- cyclohexyl ester 
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
                   
               
                 Bromo-acetic acid 3-{2-[1-(5-ethyl-5-hydroxy- 1-methyl-hept-3-ynyl)-7a-methyl-octahydro- inden-4-ylidene]-ethylidene}-4-methylene- cyclohexyl ester 
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
                   
               
                 Bromo-acetic acid 3-{2-[1-(3,5-dihydroxy-1,5- dimethyl-4-oxo-hexyl)-7a-methyl-octahydro- inden-4-ylidene]-ethylidene}-4-methylene- cyclohexyl ester 
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
                   
               
                 Bromo-acetic acid 3-{2-[1-(5-hydroxy-1,5- dimethyl-hex-3-ynyl)-7a-methyl-3,3a,5,6,7,7a- hexahydro-inden-4-ylidene]-ethylidene}-4- methylene-cyclohexyl ester 
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
                   
               
                 Bromo-acetic acid 3-{2-[1-(5-ethyl-5-hydroxy- 1-methyl-4-oxo-heptyl)-7a-methyl-octahydro- inden-4-ylidene]-ethylidene}-4-methylene- cyclohexyl ester 
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
                   
               
                 Bromo-acetic acid 3-(2-{1-[1-(4-hydroxy-4- methyl-3-oxo-pentyl)-cyclopropyl]-7a-methyl- octahydro-inden-4-ylidene}-ethylidene)-4- methylene-cyclohexyl ester 
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
                   
               
                 Bromo-acetic acid 4-methylene-3-{2-[7a- methyl-1-(6,6,6-trifluoro-5-hydroxy-1-methyl- 5-trifluoromethyl-hexyl)-octahydro-inden-4- ylidene]-ethylidene}-cyclohexyl ester 
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
                   
               
                 3-(4-{2-[5-(2-Bromo-acetoxy)-2-methylene- cyclohexylidene]-ethylidene}-7a-methyl- octahydro-inden-1-yl)-butyric acid 
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
                   
               
           
              
             
             
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
             
          
         
       
     
   
   
       23 . The method of  claim 1  further comprising obtaining the calcidiol derivative. 
   
   
       24 . A novel calcidiol derivative of formula I: 
     
       
         
         
             
             
         
       
       wherein: 
       A 1  is single or double bond; 
       A 2  is a single, double or triple bond; 
       X 1  and X 2  are each independently H 2  or CH 2 , provided X 1  and X 2  are not both ═CH 2 ;
 and X 1  can be substituted with CH 2 OR 11    
 wherein R 11  is C(O)alkyl, C(O)aryl, or C(O)aralkyl; 
 
       R 1  is H, OH, or OSi(R 10 ) 3 ;
 Wherein R 10  is alkyl, aryl, alkenyl, or aralkyl; 
 
       R 2  is O(CH 2 ) n R 12 ; 
       Wherein R 12  is halogen, haloalkyl, amino, alkyl amino, thiol, alkyl thio, hydroxyl, alkoxy, or alkenyl; 
       n is an integer from 1-6; 
       and any of the CH 2  groups may be replaced by CO; 
       R 3  and R 4  independently H, C 1 -C 4  alkyl, hydroxyalkyl, or haloalkyl, or R 3  and R 4  taken together with C 20  form C 3 -C 6  cycloalkyl; 
       R 5  is H, C 1 -C 4  alkyl, hydroxyalkyl, haloalkyl, or carbonyl; 
       R 6  and R 7  are each independently alkyl or haloalkyl;
 R 8  is H or biotin; 
 R 9  is H, hydroxyl, or halogen; and 
 
       pharmaceutically acceptable esters, salts, solvates, clathrates, stereoisomers, enantiomers prodrugs thereof. 
     
   
   
       25 . The novel calcidiol derivative of  claim 24  having formula II 
     
       
         
         
             
             
         
       
       wherein X is F, Cl, Br, I, N 3 , N 2 , NH 2 , CN, S—CH 3 , or N═C═S; 
       R is 
     
     
       
         
         
             
             
         
       
       or a pharmaceutically acceptable salt, solvate, clathrate, stereoismoer, enantiomer or prodrug thereof. 
     
   
   
       26 . The novel calcidiol derivative of  claim 24 , which is bromoacetoxycalcidiol (B3CD)(bromo-acetic acid 3-{2-[1-(5-hydroxy-1,5-dimethyl-hexyl)-7a-methyl-octahydro-inden-4-ylidene]-ethylidene}-4-methylene-cyclohexyl ester): 
     
       
         
         
             
             
         
       
       or a pharmaceutically acceptable salt, solvate, clathrate, stereoisomer, enantiomer, or prodrug thereof. 
     
   
   
       27 . The novel calcidiol derivative of  claim 24 , which is selected from the group consisting of: 
     
       
         
               
               
             
                   
               
                 Bromoacetoxycalcidiol (B3CD) Bromo-acetic acid 3-{2-[1-(5-hydroxy-1,5- dimethyl-hexyl)-7a-methyl-octahydro-inden-4- ylidene]-ethylidene}-4-methylene-cyclohexyl ester 
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
                   
               
                 25-Biotin-Bromoacetoxycalcidiol 
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
                   
               
                 6-(2,4-Dinitro-phenylamino)-hexanoic acid 2- (4-(2-bromo-acetoxy)-2-{2-[1-(5-hydroxy-1,5- dimethyl-hexyl)-7a-methyl-octahydro-inden-4- ylidene]-ethylidene}-cyclohexylidene)-ethyl ester 
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
                   
               
                 Bromo-acetic acid 3-hydroxy-5-{2-[1-(5- hydroxy-1,5-dimethyl-hexyl)-7a-methyl- octahydro-inden-4-ylidene]-ethylidene}-4- methylene-cyclohexyl ester 
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
                   
               
                 Bromo-acetic acid 3-(tert-butyl-dimethyl- silanyloxy)-5-{2-[1-(5-hydroxy-1,5-dimethyl- hexyl)-7a-methyl-octahydro-inden-4-ylidene]- ethylidene}-4-methylene-cyclohexyl ester 
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
                   
               
                 Trifluoro-acetic acid 3-{2-[1-(5-hydroxy-1,5- dimethyl-hexyl)-7a-methyl-octahydro-inden-4- ylidene]-ethylidene}-4-methylene-cyclohexyl ester 
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
                   
               
                 4-Bromo-butyric acid 3-{2-[1-(5-hydroxy-1,5- dimethyl-hexyl)-7a-methyl-octahydro-inden-4- ylidene]-ethylidene}-4-methylene-cyclohexyl ester 
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
                   
               
                 4-Cyano-butyric acid 3-{2-[1-(5-hydroxy-1,5- dimethyl-hexyl)-7a-methyl-octahydro-inden-4- ylidene]-ethylidene}-4-methylene-cyclohexyl ester 
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
                   
               
                 Acrylic acid 3-{2-[1-(5-hydroxy-1,5-dimethyl- hexyl)-7a-methyl-octahydro-inden-4-ylidene]- ethylidene}-4-methylene-cyclohexyl ester 
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
                   
               
                 6-{4-[2-(5-Bromomethoxymethoxy-2- methylene-cyclohexylidene)-ethylidene]-7a- methyl-octahydro-inden-1-yl}-2-methyl-heptan- 2-ol 
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
                   
               
                 6-{4-[2-(5-Aminomethoxymethoxy-2- methylene-cyclohexylidene)-ethylidene]-7a- methyl-octahydro-inden-1-yl}-2-methyl-heptan- 2-ol 
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
                   
               
                 6-{4-[2-(5-Bromomethoxy-2-methylene- cyclohexylidene)-ethylidene]-7a-methyl- octahydro-inden-1-yl}-2-methyl-heptan-2-ol 
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
                   
               
                 2-Methyl-6-{7a-methyl-4-[2-(2-methylene-5- ethylsulfanylmethoxy-cyclohexylidene)- ethylidene]-octahydro-inden-1-yl}-heptan-2-ol 
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
                   
               
                 Bromo-acetic acid 3-(2-{1-[2-(2-hydroxy-2- methyl-propoxy)-1-methyl-ethyl]-7a-methyl- octahydro-inden-4-ylidene}-ethylidene)-4- methylene-cyclohexyl ester 
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
                   
               
                 Bromo-acetic acid 3-{2-[1-(5-hydroxy-1,5- dimethyl-hex-3-ynyl)-7a-methyl-octahydro- inden-4-ylidene]-ethylidene}-4-methylene- cyclohexyl ester 
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
                   
               
                 Bromo-acetic acid 3-{2-[1-(5-hydroxy-1,5- dimethyl-4-oxo-hexyl)-7a-methyl-octahydro- inden-4-ylidene]-ethylidene}-4-methylene- cyclohexyl ester 
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
                   
               
                 Bromo-acetic acid 3-{2-[1-(5-ethyl-5-hydroxy- 1-methyl-hept-3-ynyl)-7a-methyl-octahydro- inden-4-ylidene]-ethylidene}-4-methylene- cyclohexyl ester 
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
                   
               
                 Bromo-acetic acid 3-{2-[1-(3,5-dihydroxy-1,5- dimethyl-4-oxo-hexyl)-7a-methyl-octahydro- inden-4-ylidene]-ethylidene}-4-methylene- cyclohexyl ester 
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
                   
               
                 Bromo-acetic acid 3-{2-[1-(5-hydroxy-1,5- dimethyl-hex-3-ynyl)-7a-methyl-3,3a,5,6,7,7a- hexahydro-inden-4-ylidene]-ethylidene}-4- methylene-cyclohexyl ester 
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
                   
               
                 Bromo-acetic acid 3-{2-[1-(5-ethyl-5-hydroxy- 1-methyl-4-oxo-heptyl)-7a-methyl-octahydro- inden-4-ylidene]-ethylidene}-4-methylene- cyclohexyl ester 
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
                   
               
                 Bromo-acetic acid 3-(2-{1-[1-(4-hydroxy-4- methyl-3-oxo-pentyl)-cyclopropyl]-7a-methyl- octahydro-inden-4-ylidene}-ethylidene)-4- methylene-cyclohexyl ester 
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
                   
               
                 Bromo-acetic acid 4-methylene-3-{2-[7a- methyl-1-(6,6,6-trifluoro-5-hydroxy-1-methyl- 5-trifluoromethyl-hexyl)-octahydro-inden-4- ylidene]-ethylidene}-cyclohexyl ester 
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
                   
               
                 3-(4-{2-[5-(2-Bromo-acetoxy)-2-methylene- cyclohexylidene]-ethylidene}-7a-methyl- octahydro-inden-1-yl)-butyric acid 
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
                   
               
           
              
             
             
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
              
             
          
         
       
     
   
   
       28 . A pharmaceutical composition comprising the calcidiol derivative of  claim 24  and a pharmaceutically acceptable carrier. 
   
   
       29 . The pharmaceutical composition  claim 28  further comprising a biologically active agent selected from the group consisting of an immunosuppressant, analgesic and antibiotic. 
   
   
       30 . The pharmaceutical composition of  claim 29 , wherein the calcidiol derivative is bromoacetoxycalcidiol (B3CD). 
   
   
       31 . The pharmaceutical composition of  claim 28  comprising a tablet, troche or liquid. 
   
   
       32 . The composition of  claim 28  comprising an effective amount of a compound of formula I: 
     
       
         
         
             
             
         
       
       or a pharmaceutically acceptable salt, solvate, clathrate, stereoisomer, enantiomer, or prodrug thereof. 
     
   
   
       33 . A packaged formulation for use in inhibiting cancer cell proliferation comprising the pharmaceutical composition of  claim 28  and instructions for use in accordance with the method of  claim 1 .

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