PATENT DOCUMENT

Abstract:
Disclosed are neuroblastoma tumor-initiating cell inhibiting compositions comprising chemical entities capable of affecting neuroblastoma tumor-initiating cells. Pharmaceutical preparations that include these chemical entities are also provided for the treatment of neuroblastoma. These pharmaceutical preparations are suitable for the treatment of humans, and are particularly suited for the treatment of children of 12 years of age or younger having neuroblastoma. The compositions and pharmaceutical preparations posses reduced normal cell cytotoxicity. The compositions and pharmaceutical preparations may be used alone or together with other conventional neuroblastoma preparations as part of a clinical regimen in the treatment and management of neuroblastoma.

Full Description:
CROSS-REFERENCE TO RELATED APPLICATIONS 
       [0001]    This application makes reference to the following co-pending U.S. patent application. The application is U.S. App. No. 60/739,337, entitled “Cancer Stem Cells and Uses Thereof,” filed Nov. 23, 2005. The entire disclosure and contents of the above application is hereby incorporated by reference. 
     
    
     BACKGROUND 
       [0002]    1. Field of the Invention 
         [0003]    The present invention relates to the field of pharmacologically active chemical compositions useful in affecting neuroblastoma tumor-initiating cells, and the use of such compositions in the treatment of neuroblastoma and related conditions. 
         [0004]    2. Related Art 
         [0005]    Neuroblastoma (NB) is the most common extracranial solid tumor in children, with poor survival rates in children with metastatic disease. NB is estimated to be responsible for about 15% of cancer-related deaths in children (1, 2). The survival rate for metastatic NB is estimated to be less than 30%. In the majority of these cases, conventional cancer therapies have been ineffective. 
         [0006]    Little is reported concerning the precise molecular alterations that give rise to NB, its cell of origin, or why NB cells metastasize and become resistant to chemotherapeutic agents. Unfortunately, genetic mutations that contribute to the origin and progression of 98% of NB cases have not been identified. 
         [0007]    One identifiable hallmark of NB is the appearance of proliferating cells with characteristics of neural crest-derived sympathetic neuronal precursors (neuroblasts). NB tumors also frequently contain other neural crest cell types, including neuroendocrine and Schwann cells. Moreover, NB appears in tissues that developmentally derive from the neural crest including sympathoadrenal precursors which differentiate into both sympathetic neurons and adrenal chromaffin cells, the paravertebral and preaortic sympathetic ganglia, and the adrenal gland. 
         [0008]    The clinical behavior of NB is unique. Tumors that arise in children under one year of age may spontaneously regress by differentiation or apoptosis, even after arising in or metastasizing to liver and skin. In contrast, NB tumors in children over one-year-old often grow aggressively, disseminate to the bone and bone marrow, and are fatal in the vast majority of cases. 
         [0009]    Mass screening of infants showed that NB is much more frequent than previously thought. Many of these tumors regress without clinical diagnosis. Regressing or favorable-prognosis tumors have been reported to express high levels of the TrkA/NGF receptor and display phenotypes of differentiated peripheral neural cells, while malignant or unfavorable-prognosis tumors resemble proliferating sympathoadrenal precursors, often expressing TrkB, amplified N-myc, and many genes involved in neural crest development. 
         [0010]    The only reported germline NB predisposition gene is Phox2b, which is mutated in many familial cases of NB, and is required for proper differentiation of sympathetic neurons from neural crest precursors (NCPs) (3, 4). In the regressive form of the disease, the transformed precursors ultimately differentiate or die, while in older children, these molecular transformations instead result in a population of persistently proliferating and highly migratory transformed neuroblasts. 
         [0011]    The concept of tumor-initiating cells (TIC) (also called tumor or cancer stem cells) postulates that only rare cells in tumors are endowed with tumorigenic potential, and was initially developed to explain why (i) most tumors are comprised of both undifferentiated proliferating progenitors and post mitotic differentiated cells, (ii) only a very small fraction of tumor cells form colonies after plating in vitro, and (iii) large numbers of tumor cells are required to seed the growth of a new tumor in mice (4-10). 
         [0012]    Dick et al. and others reported that clonally-derived tumor cells of acute myelogenous leukemia (AML) patients could be physically separated into tumorigenic and non-tumorigenic fractions (11, 12). Brain and breast tumors have also been reported to contain a subpopulation of TICs (13, 14). Thus, in solid tumors, a rare tumor cell population may fuel tumor growth and seed metastasis. This hypothesis has major implications for treating cancer patients. For example, many current therapies kill the bulk of proliferating tumor cells, but these cells may not be intrinsically tumorigenic, and in many cases the TICs may escape the effects of the therapeutic agents, leading to tumor relapse. Thus, it is essential to identify and characterize TICs from various tumors in order to develop and target therapies against this critical cell type. 
         [0013]    TICs have also been shown to share phenotypic characteristics with stem cells derived from their tissue of origin. For example, for a given tissue, the tissue stem cells and TICs both (i) self-renew, (ii) express common phenotypic markers, (iii) grow in a similar fashion in response to mitogens, and (iv) yield tissue-appropriate progeny (13, 14). However, whereas tissue stem cells generate mature differentiated cell types, differentiation of TICs is generally arrested at the level of one or more tissue progenitor cells resulting in tumors comprising a hierarchy of progenitors and some differentiated progeny (4). 
         [0014]    Many pediatric and adult tissues contain resident stem cells (4). It is currently unknown if TICs originate by transformation of tissue stem cells. Observations have been made that oncogenic mutations commonly affect genes required for normal stem cell renewal and differentiation (4). This may be particularly relevant for children&#39;s tumors, since developing tissues contain a higher proportion of tissue stem cells than do adult tissues. 
         [0015]    Tumor initiating cells from some solid tissue tumors, such as breast and brain tumors, have been described. However, a tumor initiating cell population from tumor tissue in a patient with neuroblastoma has not been isolated. One reported observation in some infantile forms of NB (called stage  4 S) is that large tumors are frequently found in skin (15). It was previously assumed that skin was a preferred metastatic target for NB. However, a population of tumor initiating cells from such solid tumor tissue has not yet been reported. 
         [0016]    The above and other observations in the field reveal a continuing medical need continues to exist in the art to determine why and in which cell type NB arises, and why some neuroblastoma tumors spontaneously regress and others are fatal. In addition, new effective drug targets and therapeutics tailored to identifying and treating specific forms and stages of neuroblastoma are needed. 
       SUMMARY 
       [0017]    The above and other long-felt needs in the art are met in the present invention. 
       Compositions/Pharmaceutical Preparations: 
       [0018]    In one aspect, the invention provides compositions comprising novel chemical entities that are capable of affecting neuroblastoma. In some embodiments, these chemical entities may be described as compounds that specifically kill neuroblastoma tumor-initiating cells, or that arrest the growth of neuroblastoma tumor-initiating cells. In other aspects, these chemical entities and compositions containing one or more of them may be described as having specifically cytostatic or cytotoxic activity toward neuroblastoma tumor-initiating cells. 
         [0019]    In some embodiments, the anti-neuroblastoma composition may be described as comprising one or more active ingredients comprising: 
         [0020]    2.3-Dimethoxy-1.4-naphthoquinone, 
         [0021]    Aklavine Hydrochloride, 
         [0022]    Amodiaquin dihydrochloride dehydrate; 
         [0023]    Amsacrine Hydrochloride; 
         [0024]    Azaguanine-8; 
         [0025]    beta-peltatin; 
         [0026]    Camptothecine (S.+); 
         [0027]    CGP-74514A hydrochloride; 
         [0028]    Chelerythrine chloride; 
         [0029]    Cholestan-3beta.5alpha.6beta-Triol; 
         [0030]    Ciclopirox Olamine; 
         [0031]    Clofazimine; 
         [0032]    Colchicine; 
         [0033]    Convallatoxin; 
         [0034]    Crassin Acetate; 
         [0035]    Crinamine; 
         [0036]    Dequalinium analog. C-14 linker; 
         [0037]    Dequalinium dichloride; 
         [0038]    Digitoxin; 
         [0039]    Digoxigenin; 
         [0040]    Dihydrogambogic acid; 
         [0041]    Dihydroouabain; 
         [0042]    Erysolin; 
         [0043]    Gambogic acid; 
         [0044]    Mechlorethamine; 
         [0045]    Meclizine hydrochloride; 
         [0046]    MG 624; 
         [0047]    Mitoxanthrone Hydrochloride; 
         [0048]    Ouabain; 
         [0049]    Oxybendazole; 
         [0050]    Oxybendazole; 
         [0051]    Paclitaxel; 
         [0052]    Parthenolide; 
         [0053]    Patulin; 
         [0054]    Periplocymarin; 
         [0055]    Peruvoside; 
         [0056]    Primaquine diphosphate; 
         [0057]    Quinacrine dihydrochloride; 
         [0058]    Sanguinarine chloride; or 
         [0059]    Tomatine, 
         [0060]    In some embodiments, the chemical entities of the invention may be described as compounds that possess specific cytostatic or cytotoxic activity toward neuroblastoma tumor-initiating cells. In other embodiments, the neuroblastoma-inhibiting composition further comprises ancitabine hydrochloride, doxorubicin hydrochloride, etoposide, vincristine sulfate, or a combination thereof. In yet other embodiments, the neuroblastoma inhibiting composition may be further described as having reduced non-neuroblastoma tumor-initiating cell cytotoxicity. 
         [0061]    In some embodiments, the chemical entities may be described as possessing potent anti-neuroblastoma tumor-initiating cell activity, and a reduced cytotoxicity to normal, non-neuroblastoma tumor-initiating cells, relative to conventional neuroblastoma treatment preparations. In some embodiments, the compositions are described as essentially free of non-neuroblastoma tumor cell inhibiting activity. 
         [0000]    Methods of Inhibiting Neuroblastoma Tumor initiating Cells/Methods of Treating and/or Inhibiting Neuroblastoma in an Animal 
         [0062]    In yet another aspect, the invention provides methods for inhibiting neuroblastoma tumor-initiating cells. In some embodiments, the method comprises administering an effective amount of a composition comprising a neuroblastoma tumor-initiating cell inhibiting ingredient. In some embodiments, the neuroblastoma tumor-initiating cell inhibiting ingredient comprises one or more active ingredients comprising: 
         [0063]    2,3-Dimethoxy-1,4-naphthoquinone, 
         [0064]    Aklavine Hydrochloride, 
         [0065]    Amodiaquin dihydrochloride dehydrate; 
         [0066]    Amsacrine Hydrochloride; 
         [0067]    Azaguanine-8; 
         [0068]    beta-peltatin; 
         [0069]    Camptothecine (S.+); 
         [0070]    CGP-74514A hydrochloride; 
         [0071]    Chelerythrine chloride; 
         [0072]    Cholestan-3beta.5alpha.6beta-Triol; 
         [0073]    Ciclopirox Olamine; 
         [0074]    Clofazimine; 
         [0075]    Colchicine; 
         [0076]    Convallatoxin; 
         [0077]    Crassin Acetate; 
         [0078]    Crinamine; 
         [0079]    Dequalinium analog. C-14 linker; 
         [0080]    Dequalinium dichloride; 
         [0081]    Digitoxin; 
         [0082]    Digoxigenin; 
         [0083]    Dihydrogambogic acid; 
         [0084]    Dihydroouabain; 
         [0085]    Erysolin; 
         [0086]    Gambogic acid; 
         [0087]    Mechlorethamine; 
         [0088]    Meclizine hydrochloride; 
         [0089]    MG 624; 
         [0090]    Mitoxanthrone Hydrochloride; 
         [0091]    Ouabain; 
         [0092]    Oxybendazole; 
         [0093]    Oxybendazole; 
         [0094]    Paclitaxel; 
         [0095]    Parthenolide; 
         [0096]    Patulin; 
         [0097]    Periplocymarin; 
         [0098]    Peruvoside; 
         [0099]    Primaquine diphosphate; 
         [0100]    Quinacrine dihydrochloride; 
         [0101]    Sanguinarine chloride; or 
         [0102]    Tomatine. 
         [0103]    In some embodiments, the effective amount of the neuroblastoma tumor initiating cell inhibiting ingredient is an amount effective to arrest the growth of and/or kill neuroblastoma tumor-initiating cells, or effective to induce differentiation of said cells to cell types that no longer proliferate. In other embodiments, the method may further comprise administering a composition further comprising ancitabine hydrochloride, doxorubicin hydrochloride, etoposide, vincristine sulfate, or a combination thereof. 
         [0104]    In some embodiments, the composition may further include a pharmaceutically acceptable carrier solution. 
         [0105]    In yet other embodiments, the neuroblastoma tumor-inhibiting cells are in an animal having neuroblastoma. In some embodiments, the animal is a human. In some embodiments, the human is 12 years of age or younger. That is, it is anticipated that the invention is particularly useful in the treatment of children afflicted with neuroblastoma, and will have a profound effect on reducing the high rate of mortality in this population of neuroblastoma patients. 
         [0106]    The method may be further described as administering a composition that has a reduced non-neuroblastoma tumor-initiating cell cytotoxicity. It is expected that the methods and compositions of the present invention will provide fewer and/or less pronounce undesirable side affect in the treatment of a patient as a result. In some embodiments, the composition employed in the method is essentially free of non-neuroblastoma tumor cell inhibiting activity. 
         [0107]    The compositions may be described as comprising a mixture of any or all of the compounds listed below: 
         [0000]    
       
         
               
               
             
           
               
                   
                 TABLE 1 
               
               
                   
                   
               
             
             
               
                   
                 2.3-Dimethoxy-1.4-naphthoquinone 
               
               
                   
                 AKLAVINE HYDROCHLORIDE 
               
               
                   
                 Amodiaquin dihydrochloride dihydrate 
               
               
                   
                 AMSACRINE HYDROCHLORIDE 
               
               
                   
                 ANCITABINE HYDROCHLORIDE 
               
               
                   
                 Azaguanine-8 
               
               
                   
                 beta-PELTATIN 
               
               
                   
                 Camptothecine (S.+) 
               
               
                   
                 CGP-74514A hydrochloride 
               
               
                   
                 Chelerythrine chloride 
               
               
                   
                 CHOLESTAN-3beta.5alpha.6beta-TRIOL 
               
               
                   
                 CICLOPIROX OLAMINE 
               
               
                   
                 Clofazimine 
               
               
                   
                 Colchicine 
               
               
                   
                 CONVALLATOXIN 
               
               
                   
                 CRASSIN ACETATE 
               
               
                   
                 CRINAMINE 
               
               
                   
                 Dequalinium analog. C-14 linker 
               
               
                   
                 Dequalinium dichloride 
               
               
                   
                 Digitoxin 
               
               
                   
                 Digoxigenin 
               
               
                   
                 Digoxin 
               
               
                   
                 DIHYDROGAMBOGIC ACID 
               
               
                   
                 Dihydroouabain 
               
               
                   
                 ERYSOLIN 
               
               
                   
                 Etoposide 
               
               
                   
                 GAMBOGIC ACID 
               
               
                   
                 Idarubicin 
               
               
                   
                 MECHLORETHAMINE 
               
               
                   
                 MECLIZINE HYDROCHLORIDE 
               
               
                   
                 MG 624 
               
               
                   
                 MITOXANTHRONE HYDROCHLORIDE 
               
               
                   
                 OUABAIN 
               
               
                   
                 OXYBENDAZOLE 
               
               
                   
                 Paclitaxel 
               
               
                   
                 Parthenolide 
               
               
                   
                 PATULIN 
               
               
                   
                 PERIPLOCYMARIN 
               
               
                   
                 PERUVOSIDE 
               
               
                   
                 Podophyllotoxin 
               
               
                   
                 Primaquine diphosphate 
               
               
                   
                 Quinacrine dihydrochloride 
               
               
                   
                 Sanguinarine chloride 
               
               
                   
                 TENIPOSIDE 
               
               
                   
                 TOMATINE 
               
               
                   
                 Vinblastine sulfate salt 
               
               
                   
                 Vincristine sulfate 
               
               
                   
                   
               
             
          
         
       
     
     
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         [0108]      FIG. 1 , according to one embodiment of the invention, presents a diagrammatic flow chart demonstrating the design of the high throughput, dual-cell (Normal or Tumor cells) screening assay employed in the selection of candidate test compounds that target neuroblastoma tumor-initiating cells. Normal or tumor-initiating cell spheres are dissociated; 3,000 single cells/well are plated in 96 well plates; candidate test compound is added; cell proliferation assayed by Alamar Blue signal. Blue/nonfluorescent compound is converted to a red/fluorescent compound under reducing conditions such as those produced by live cells. The magnitude of the fluorescent signal is proportional to the metabolic activity of the cell sample. 
           [0109]      FIG. 2 , according to one embodiment of the invention, presents the results from a study wherein FS90 (normal human SKPs, passage 3) cells were treated with the LOPAC library of chemical compounds. Alamar Blue was added after 30 hours and fluorescence intensity read after an additional 24 hours. The hit cutoff is indicated in the graph by the thick line across the graph at the Y axis value of about 69.00% Control Alamar Blue Signal (which corresponds to 3 standard deviations from the mean of all test samples). Nine compounds whose Alamar Blue signals fall below this line were identified as primary hits in this study. (X axis presents the Compound ID number (n=80×8 plates); Y axis presents the % Control Alamar Blue Signal). 
           [0110]      FIG. 3A-3C , according to one embodiment of the invention, presents the study results from primary screens of the chemical libraries examined.  3 A presents the results of the primary screen in Venn diagram form. The Venn diagrams depict the primary hits from each library. Compounds in the gray-bordered circles (left circle) affected the tumor-initiating cells, while compounds in the black-bordered circles (right circle) affected normal cells. Compounds that affected both cell types lie in the overlap region. Note that there is some compound redundancy between the libraries.  3 B presents the confirmed primary hits in Venn diagram form. Primary hits were retested against NB12, FS90 and FS105 (normal human SKPs). 87% of the primary hits were confirmed in this step, yielding 54 unique compounds that target tumor-initiating cells, 4 unique compounds that target normal cells, and 46 compounds that have activity against both normal and tumor cells (overlap region).  3 C presents in a pie-format the classification of primary hits by mechanism of action. (Solid light gray area=DNA damaging agents/cell cycle inhibitors; Solid dark gray area=Na+/K+ATPase inhibitors; Diagonal striped area=Neuronal receptor effectors; Vertical striped area=Other; Solid white area=Metabolic inhibitor; Checkerboard area=Neuronal channel effectors; Dotted area=Specific protein effectors). 
           [0111]      FIG. 4A-4E , according to one embodiment of the invention, presents the IC50 values that were determined for the 64 selected candidate compounds. Compounds were chosen for further testing based on differential cell type selectivity, mechanism of action, and pharmacological interest. Tumor-initiating cells and normal cells were treated with 10 serial dilutions of compounds (5 μM to 9 nM). Representative graphs are shown in  FIG. 4A  (Complete Response),  4 B (Partial Response), and  4 C (Threshold Effect). Compounds that affected the tumor-initiating cells at a much lower dose than normal cells ( 4 D graph, left) or compounds that had a greater effect on the tumor-initiating cells than normal cells ( 4 E, right graph), were selected for secondary in vitro screens in addition to those compounds that only affected tumor-initiating cells. (FS90=normal cells; NB12=tumor-initiating cells). 
           [0112]      FIG. 5A-5C , according to one embodiment of the invention, presents the results from secondary screens of the candidate compounds. Compounds of interest are being tested against additional normal primary cultures (FS89, FS105), a stage IV neuroblastoma primary culture (NB25), and a neuroblastoma cell line (KCNR) using a sphere formation assay.  5 A presents a flow diagram of the secondary in vitro screen. The candidate compound is added at 0 days and at 3 days. Spheres are counted at 7 days.  FIG. 5B  presents a dose response curve of various cell lines (FS89, FS90, FS105, NB12, NB25 and KCNR) to amsacrine.  FIG. 5C  presents a dose response curve of various cell lines (FS89, FS90, FS105, NB12, NB25 and KCNR) to MG624. 
           [0113]    FIG.  6 A- 6 FF, according to one embodiment of the invention, presents IC 50  values for 32 selected compounds from the LOPAC and Prestwick collections. Tumor-initiating cells (NB12) and normal cells (FS90) were treated with 10 serial dilutions of compounds ranging from 5 μM to 9 nM. Cell survival/growth was assayed using Alamar Blue and the percentage of control Alamar Blue signal was plotted versus log[compound] nM. IC50 values for NB12 are given above each plot. 
           [0114]    FIG.  7 A- 7 FF, according to one embodiment of the invention, represents IC50 values determined for 32 selected compounds from the LOPAC, Prestwick, and Spectrum collections. Tumor-initiating cells (NB12) and normal cells (FS90) were treated with 10 serial dilutions of compounds ranging from 5 μM to 9 nM. Cell survival/growth was assayed using Alamar Blue and the percentage of control Alamar Blue signal was plotted versus log[compound] nM (FS90 in dashed line, NB12 in bolded line). IC50 values for NB12 and FS90 are given beside each plot. 
       
    
    
     DETAILED DESCRIPTION 
     Definitions 
       [0115]    Where the definition of terms departs from the commonly used meaning of the term, applicant intends to utilize the definitions provided below, unless specifically indicated. 
         [0116]    For the purposes of the present invention, “a”, “an” and “the” include reference to the plural unless the context as herein presented clearly indicates other wise. 
         [0117]    For purposes of the present invention, the term “active agent” is defined as a chemical entity, group of chemical entities or compound that is capable of providing an affect on neuroblastoma tumor initiating cells or neuroblastoma cells in vitro or in vivo. The affect of the active agent may be a reduction in cytotoxicity relative to the level of cytotoxicity demonstrated in the absence of the active agent under similar conditions, or a cytostatic affect on neuroblastoma or on neuroblastoma tumor initiating cells that results in a reduced rate of neuroblastoma or neuroblastoma tumor-initiating cell proliferation and/or growth, or a reduction of the rate or occurrence of differentiation into neuroblastoma cell types, precursors, or any other cell type that is related to the progression of a neuroblastoma pathology, or to an increase in the inducement of the differentiation of neuroblastoma tumor-initiating cells into cell types (for example, neurons) that no longer proliferate (for example, retinoic acid is a common differentiation therapy for neuroblastoma that is used as an adjunct therapy after removal of a tumor, differentiation therapy). 
         [0118]    For purposes of the present invention, the term “enriched” is defined as containing at least 50% of the identified biological moiety, such as a cancer stem cell. 
         [0119]    For purposes of the present invention, the term “neuroblastoma tumor initiating cell” (NB TIC) is defined as a cell that is capable of giving rise to neuroblastoma or a tumor cell that is identifiable with a condition of neuroblastoma, such as a tumor cell that may be identified to have particular identifiable cell surface markers associated with neuroblastoma (such as NB84, CD44, TrkA, GD2, CD34, p75NTR, and/or versican) and/or is without cell surface markers that are characteristic of tumor cells that are not from neuroblastoma (such as CD133, TrkB, and/or CD31). 
         [0120]    For purposes of the present invention, the term “neuroblastoma tumor-initiating cell inhibiting activity” is defined as an activity for affecting neuroblastoma tumor-initiating cell survival, proliferation, or that promotes cell differentiation into benign cell types. 
         [0121]    For purposes of the present invention, the term, “effective amount” is defined as an amount of a compound that will inhibit and/or reduce neuroblastoma tumor initiating cell survival, proliferation, or that will promote the differentiation of neuroblastoma tumor-initiating cells into benign cell types. 
       DESCRIPTION 
     Examples 
     Example I Materials and Methods 
       [0122]    The present example provides a description of the screening method used to identify the chemical entities capable of affecting neuroblastoma cells reported in the present series of studies. 
         [0123]    Malignant neuroblastoma (NB) is the most common extra-cranial solid tumor in children. Survival of patients older than 1 year remains less than 30% with conventional therapies. Candidate NB tumor-initiating cells (TICs) were isolated, and it was hypothesized that TICs are related to SKPs (SKin-derived Precursors). Both SKPs and TICs originate from the neural crest, express similar neural crest markers, and differentiate in vitro into similar cell types. The availability of two neural crest stem cell sources, one from the NB tumor and the other from the skin of the same patient, affords us a unique opportunity for therapeutic target discovery. 
       Study 1 Screen: 
     Materials and Methods: 
       [0124]    To identify compounds that suppress the growth and survival of neuroblastoma (NB) tumor-initiating cells and not nontransformed normal cells (SKPs), a cell-based assay was established and used in which NB tumor-initiating cells from a multiple relapse NB patient (NB12, passage 6-17) and normal SKPs (FS90, passage 2-5) were tested in parallel to detect specific alterations of cell viability/proliferation. For each cell type, cells were passaged 5 days prior to screening. Three thousand (3000) cells in 100□L SKPs growth media (B27, bFGF, EGF, P/S, fungizone in DMEM:F12 with 50% hFS conditioned media) were robotically plated in uncoated 96 well plates and treated with test compound for 30 hours, prior to a 24 hour incubation in the presence of Alamar Blue and subsequent fluorometric reading. Under these conditions, the Alamar Blue signal displayed a linear response with time, background was minimal, and the dynamic range satisfactory (i.e. the Alamar Blue reading at 0 hours vs. 24 hours was &gt;10 fold different). 
         [0125]    The robustness of the screen was initially evaluated by using a collection of 1280 bioactive compounds (LOPAC library, Sigma). For both normal SKPs and NB tumor-initiating cells, variability of signals was low, with CV values ranging between 3.5-4.5% across the plates, and the dimensionless, statistical parameters Z′ and Z factors were &gt;0.5, suggesting an excellent assay quality. “Hits” were defined as the compounds whose signals were shifted away by at least 3×standard deviations (99.73% confidence interval) from the mean of the general sample population. 
       Results: 
       [0126]    The screen of the LOPAC library at 5 μM yielded 13 “hits” which were found to affect both normal and NB cells. We also identified 18 compounds that selectively target NB cells. Four compounds selectively targeted normal cells. 
         [0000]    
       
         
               
             
               
               
             
           
               
                 TABLE 2 
               
               
                   
               
               
                 13 compounds that affect both normal and NB cells: 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Ancitabine hydrochloride 
               
               
                   
                 Brefeldin A from Penicillium brefeldianum 
               
               
                   
                 Calmidazolium chloride 
               
               
                   
                 CGP-74514A hydrochloride 
               
               
                   
                 Dihydroouabain 
               
               
                   
                 Diphenyleneiodonium chloride 
               
               
                   
                 Emetine dihydrochloride hydrate 
               
               
                   
                 Idarubicin 
               
               
                   
                 Mitoxantrone 
               
               
                   
                 Ouabain 
               
               
                   
                 Quinacrine dihydrochloride 
               
               
                   
                 Ammonium pyrrolidinedithiocarbamate 
               
               
                   
                 Sanguinarine chloride 
               
               
                   
                   
               
             
          
         
       
     
         [0000]    
       
         
               
             
               
               
             
           
               
                 TABLE 3 
               
               
                   
               
               
                 18 compounds that selectively target NB cells. 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Loratadine 
               
               
                   
                 MG 624 
               
               
                   
                 Melphalan 
               
               
                   
                 Podophyllotoxin 
               
               
                   
                 Ro 25-6981 hydrochloride 
               
               
                   
                 Rotenone 
               
               
                   
                 DL-Stearoylcarnitine chloride 
               
               
                   
                 Taxol 
               
               
                   
                 Vincristine sulfate 
               
               
                   
                 Vinblastine sulfate salt 
               
               
                   
                 Chelerythrine chloride 
               
               
                   
                 Colchicine 
               
               
                   
                 Cytosine-1-beta-D-arabinofuranoside hydrochloride 
               
               
                   
                 Dequalinium dichloride 
               
               
                   
                 (S)-(+)-Camptothecin 
               
               
                   
                 Dequalinium analog, C-14 linker 
               
               
                   
                 2,3-Dimethoxy-1,4-naphthoquinone 
               
               
                   
                 Etoposide 
               
               
                   
                   
               
             
          
         
       
     
         [0000]    
       
         
               
             
               
               
             
           
               
                 TABLE 4 
               
               
                   
               
               
                 4 compounds selectively target normal cells: 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 8-Methoxymethyl-3-isobutyl-1-methylxanthine 
               
               
                   
                 Oligomycin A 
               
               
                   
                 Sphingosine 
               
               
                   
                 Thapsigargin 
               
               
                   
                   
               
             
          
         
       
     
       Study 2 Screen: 
     Materials and Methods 
       [0127]    The Prestwick library was screened at 5 μM using FS90 and NB12 and at 1 μM using NB12 only due to the high number of “hits” at 5M. This screen identified 9 compounds that selectively target NB12 and 15 compounds that affect both NB12 and FS90. 
       Results: 
       [0128]      
         [0000]    
       
         
               
             
               
               
             
           
               
                 TABLE 5 
               
               
                   
               
               
                 9 compounds that selectively target NB12: 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Azaguanine-8 
               
               
                   
                 Paclitaxel 
               
               
                   
                 Camptothecine (S.+) 
               
               
                   
                 Colchicine 
               
               
                   
                 Etoposide 
               
               
                   
                 Doxorubicin hydrochloride 
               
               
                   
                 Lanatoside C 
               
               
                   
                 Podophyllotoxin 
               
               
                   
                 Proscillaridin A 
               
               
                   
                   
               
             
          
         
       
     
         [0000]    
       
         
               
             
               
               
             
           
               
                 TABLE 6 
               
               
                   
               
               
                 15 compounds that affect both NB12 and FS90: 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Disulfiram 
               
               
                   
                 Mitoxantrone dihydrochloride 
               
               
                   
                 Anisomycin 
               
               
                   
                 Cephaeline dihydrochloride heptahydrate 
               
               
                   
                 Digitoxigenin 
               
               
                   
                 Digoxin 
               
               
                   
                 Strophantine octahydrate 
               
               
                   
                 Puromycin dihydrochloride 
               
               
                   
                 Daunorubicin hydrochloride 
               
               
                   
                 Emetine dihydrochloride 
               
               
                   
                 Methyl benzethonium chloride 
               
               
                   
                 Strophanthidin 
               
               
                   
                 Cycloheximide 
               
               
                   
                 Thonzonium bromide 
               
               
                   
                 Sanguinarine 
               
               
                   
                   
               
             
          
         
       
     
       Study 3 Screen: 
     Methods: 
       [0129]    The results from the LOPAC and Prestwick screens were confirmed using FS90, FS105, and NB12. Thirty-six (36) compounds were confirmed that specifically affect NB12 and 29 compounds that affect FS90/105 and NB12. Thirty-two (32) compounds were selected for IC 50  determinations using FS90, FS105, and NB12. IC 50  for the remaining compounds of interest will be tested at a later date (in combination with hits from additional libraries). 
       Results: 
       [0130]      
         [0000]    
       
         
               
             
               
               
             
           
               
                 TABLE 7 
               
               
                   
               
               
                 36 compounds that specifically affect NB12 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 (S)-(+)-Camptothecin 
               
               
                   
                 2.3-Dimethoxy-1.4-naphthoquinone 
               
               
                   
                 Ancitabine hydrochloride 
               
               
                   
                 Antimycin A 
               
               
                   
                 Azaguanine-8 
               
               
                   
                 Benzethonium chloride 
               
               
                   
                 Camptothecine (S.+) 
               
               
                   
                 Chelerythrine chloride 
               
               
                   
                 Ciclopirox ethanolamine 
               
               
                   
                 Clofazimine 
               
               
                   
                 Colchicine 
               
               
                   
                 Colchicine 
               
               
                   
                 Cycloheximide 
               
               
                   
                 Cytosine-1-beta-D-arabinofuranoside 
               
               
                   
                 hydrochloride 
               
               
                   
                 Dequalinium analog. C-14 linker 
               
               
                   
                 Dequalinium dichloride 
               
               
                   
                 Dequalinium dichloride 
               
               
                   
                 Digoxigenin 
               
               
                   
                 Diphenyleneiodonium chloride 
               
               
                   
                 DL-Stearoylcarnitine chloride 
               
               
                   
                 Doxorubicin hydrochloride 
               
               
                   
                 Etoposide 
               
               
                   
                 Etoposide 
               
               
                   
                 MG 624 
               
               
                   
                 Mycophenolic acid 
               
               
                   
                 Paclitaxel 
               
               
                   
                 Parthenolide 
               
               
                   
                 Podophyllotoxin 
               
               
                   
                 Podophyllotoxin 
               
               
                   
                 Primaquine diphosphate 
               
               
                   
                 Quinacrine dihydrochloride 
               
               
                   
                 Quinacrine dihydrochloride dihydrate 
               
               
                   
                 Scoulerine 
               
               
                   
                 Taxol 
               
               
                   
                 Vinblastine sulfate salt 
               
               
                   
                 Vincristine sulfate 
               
               
                   
                   
               
             
          
         
       
     
         [0000]    
       
         
               
             
               
               
             
           
               
                 TABLE 8 
               
               
                   
               
               
                 29 compounds that affect FS90/105 and NB12 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Alexidine dihydrochloride 
               
               
                   
                 Ammonium 
               
               
                   
                 pyrrolidinedithiocarbamate 
               
               
                   
                 Amodiaquin dihydrochloride 
               
               
                   
                 dihydrate 
               
               
                   
                 Anisomycin 
               
               
                   
                 Brefeldin A from Penicillium 
               
               
                   
                 brefeldianum 
               
               
                   
                 Calmidazolium chloride 
               
               
                   
                 Cephaeline dihydrochloride 
               
               
                   
                 heptahydrate 
               
               
                   
                 CGP-74514A hydrochloride 
               
               
                   
                 Daunorubicin hydrochloride 
               
               
                   
                 Digitoxigenin 
               
               
                   
                 Digoxin 
               
               
                   
                 Dihydroouabain 
               
               
                   
                 Disulfiram 
               
               
                   
                 Emetine dihydrochloride 
               
               
                   
                 Emetine dihydrochloride hydrate 
               
               
                   
                 Idarubicin 
               
               
                   
                 Lanatoside C 
               
               
                   
                 Methyl benzethonium chloride 
               
               
                   
                 Mitoxantrone 
               
               
                   
                 Mitoxantrone dihydrochloride 
               
               
                   
                 Ouabain 
               
               
                   
                 Proscillaridin A 
               
               
                   
                 Puromycin dihydrochloride 
               
               
                   
                 Sanguinarine 
               
               
                   
                 Sanguinarine chloride 
               
               
                   
                 Strophanthidin 
               
               
                   
                 Strophantine octahydrate 
               
               
                   
                 Terfenadine 
               
               
                   
                 Thonzonium bromide 
               
               
                   
                   
               
             
          
         
       
     
         [0000]    
       
         
               
             
               
               
             
           
               
                 TABLE 9 
               
               
                   
               
               
                 32 compounds selected for IC50 determinations using FS90, FS105, 
               
               
                 and NB12: 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 (S)-(+)-Camptothecin 
               
               
                   
                 Ammonium pyrrolidinedithiocarbamate 
               
               
                   
                 Amodiaquin dihydrochloride dihydrate 
               
               
                   
                 Antimycin A 
               
               
                   
                 Avermectin B1 
               
               
                   
                 Azaguanine-8 
               
               
                   
                 Chelerythrine chloride 
               
               
                   
                 Clofazimine 
               
               
                   
                 Colchicine 
               
               
                   
                 Dequalinium analog, C-14 linker 
               
               
                   
                 Dequalinium dichloride (LOPAC 
               
               
                   
                 compound) 
               
               
                   
                 Dequalinium dichloride (Prestwick 
               
               
                   
                 compound) 
               
               
                   
                 Digoxin 
               
               
                   
                 Dihydroouabain 
               
               
                   
                 Diphenyleneiodonium chloride 
               
               
                   
                 DL-Stearoylcarnitine chloride 
               
               
                   
                 Etoposide 
               
               
                   
                 Idarubicin 
               
               
                   
                 Loratadine 
               
               
                   
                 MG 624 
               
               
                   
                 Myophenolic Acid 
               
               
                   
                 Paclitaxel 
               
               
                   
                 Parthenolide 
               
               
                   
                 Podophyllotoxin 
               
               
                   
                 Primaquine diphosphate 
               
               
                   
                 Quinacrine dihydrochloride 
               
               
                   
                 Sanguinarine chloride 
               
               
                   
                 Scoulerine 
               
               
                   
                 Strophanthidin 
               
               
                   
                 Terfenadine 
               
               
                   
                 Vinblastine sulfate salt 
               
               
                   
                 Vincristine sulfate 
               
               
                   
                   
               
             
          
         
       
     
       Study 4—Screen Results at 5 μM: 
       [0131]    The Spectrum collection was screened using the same protocol. At 5 μM, the initial screen identified 35 hits that affect NB12 and FS90, no hits that specifically target FS90, and 41 hits that specifically target NB12. The screen was repeated at 5 μM and 1 μM using NB12 and FS90 to confirm these hits and identified 34 hits that affect NB12 and FS90, no hits that specifically target FS90, and 33 hits that specifically target NB12. Following the Spectrum confirmatory screen, IC 50  determinations for an additional 32 compounds were performed. 
         [0000]    
       
         
               
             
               
               
             
           
               
                 TABLE 10 
               
               
                   
               
               
                 34 hits that affect NB12 and FS90: 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 3-METHYLORSELLINIC ACID 
               
               
                   
                 5alpha-CHOLESTAN-3beta-OL-6-ONE 
               
               
                   
                 5-AZACYTIDINE 
               
               
                   
                 AKLAVINE HYDROCHLORIDE 
               
               
                   
                 CETRIMONIUM BROMIDE 
               
               
                   
                 CHELIDONINE (+) 
               
               
                   
                 COLCHICEINE 
               
               
                   
                 COLCHICINE 
               
               
                   
                 CYTARABINE 
               
               
                   
                 DACTINOMYCIN 
               
               
                   
                 DEOXYSAPPANONE B 7.3′-DIMETHYL ETHER 
               
               
                   
                 DIGITOXIN 
               
               
                   
                 DIHYDROGAMBOGIC ACID 
               
               
                   
                 DISULFIRAM 
               
               
                   
                 EMETINE 
               
               
                   
                 GENTIAN VIOLET 
               
               
                   
                 JUGLONE 
               
               
                   
                 LANATOSIDE C 
               
               
                   
                 LYCORINE 
               
               
                   
                 MITOMYCIN C 
               
               
                   
                 OXYPHENBUTAZONE 
               
               
                   
                 PATULIN 
               
               
                   
                 PERIPLOCYMARIN 
               
               
                   
                 PERUVOSIDE 
               
               
                   
                 PHENYLMERCURIC ACETATE 
               
               
                   
                 PUROMYCIN HYDROCHLORIDE 
               
               
                   
                 PYRITHIONE ZINC 
               
               
                   
                 PYRROMYCIN 
               
               
                   
                 RETUSOQUINONE 
               
               
                   
                 SANGUINARINE SULFATE 
               
               
                   
                 SARMENTOGENIN 
               
               
                   
                 STROPHANTHIDIN 
               
               
                   
                 THIMEROSAL 
               
               
                   
                 TOMATINE 
               
               
                   
                   
               
             
          
         
       
     
         [0000]    
       
         
               
             
               
               
             
           
               
                 TABLE 11 
               
               
                   
               
               
                 33 hits that specifically target NB12: 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 10-HYDROXYCAMTOTHECIN 
               
               
                   
                 4′-DEMETHYLEPIPODOPHYLLOTOXIN 
               
               
                   
                 ANDROGRAPHOLIDE 
               
               
                   
                 AMODIAQUINE DIHYDROCHLORIDE 
               
               
                   
                 AMSACRINE HYDROCHLORIDE 
               
               
                   
                 ANCITABINE HYDROCHLORIDE 
               
               
                   
                 BENZALKONIUM CHLORIDE 
               
               
                   
                 BENZETHONIUM CHLORIDE 
               
               
                   
                 BEPRIDIL HYDROCHLORIDE 
               
               
                   
                 beta-PELTATIN 
               
               
                   
                 CAMPTOTHECIN 
               
               
                   
                 CETYLPYRIDINIUM CHLORIDE 
               
               
                   
                 CHOLESTAN-3beta.5alpha.6beta-TRIOL 
               
               
                   
                 CICLOPIROX OLAMINE 
               
               
                   
                 CONVALLATOXIN 
               
               
                   
                 CRASSIN ACETATE 
               
               
                   
                 CRINAMINE 
               
               
                   
                 DIGOXIN 
               
               
                   
                 ERYSOLIN 
               
               
                   
                 GAMBOGIC ACID 
               
               
                   
                 IMIDACLOPRIDE 
               
               
                   
                 LIMONIN 
               
               
                   
                 MECHLORETHAMINE 
               
               
                   
                 MECLIZINE HYDROCHLORIDE 
               
               
                   
                 OUABAIN 
               
               
                   
                 OXYBENDAZOLE 
               
               
                   
                 PACLITAXEL 
               
               
                   
                 PARAROSANILINE PAMOATE 
               
               
                   
                 PARTHENOLIDE 
               
               
                   
                 PODOPHYLLOTOXIN ACETATE 
               
               
                   
                 STROPHANTHIDINIC ACID LACTONE ACETATE 
               
               
                   
                 TENIPOSIDE 
               
               
                   
                 VINBLASTINE SULFATE 
               
               
                   
                   
               
             
          
         
       
     
         [0000]    
       
         
               
             
               
               
             
           
               
                 TABLE 12 
               
               
                   
               
               
                 32 Compounds selected for IC50 Determinations: 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Aklavine hydrochloride 
               
               
                   
                 AMSACRINE HYDROCHLORIDE 
               
               
                   
                 ANCITABINE HYDROCHLORIDE 
               
               
                   
                 ANDROGRAPHOLIDE 
               
               
                   
                 BEPRIDIL HYDROCHLORIDE 
               
               
                   
                 beta-PELTATIN 
               
               
                   
                 CGP-74514A hydrochloride 
               
               
                   
                 CHOLESTAN-3beta.5alpha.6beta-TRIOL 
               
               
                   
                 CICLOPIROX OLAMINE 
               
               
                   
                 CONVALLATOXIN 
               
               
                   
                 CRASSIN ACETATE 
               
               
                   
                 CRINAMINE 
               
               
                   
                 DIHYDROGAMBOGIC ACID 
               
               
                   
                 ERYSOLIN 
               
               
                   
                 Gambogic Acid 
               
               
                   
                 IMIDACLOPRIDE 
               
               
                   
                 JUGLONE 
               
               
                   
                 LIMONIN 
               
               
                   
                 MECHLORETHAMINE 
               
               
                   
                 MECLIZINE HYDROCHLORIDE 
               
               
                   
                 Mitomycin C 
               
               
                   
                 Mitoxantrone hydrochloride 
               
               
                   
                 OUABAIN 
               
               
                   
                 OXYBENDAZOLE 
               
               
                   
                 PARAROSANILINE PAMOATE 
               
               
                   
                 PERIPLOCYMARIN 
               
               
                   
                 PERUVOSIDE 
               
               
                   
                 Prenyletin 
               
               
                   
                 PYRITHIONE ZINC 
               
               
                   
                 TENIPOSIDE 
               
               
                   
                 Tomatidine hydrochloride 
               
               
                   
                 TOMATINE 
               
               
                   
                   
               
             
          
         
       
     
         [0132]    These results suggest that patient-specific therapeutics, as well as the molecular and biochemical alterations that lead to neuroblastoma, can be identified using this assay. 
       Example 2 
     Identified Compounds that Affect Normal, NB, or NB and Normal Cells 
       [0133]    The present example provides a description of the screening method used to identify and select chemical entities capable of affecting (i.e., reducing and/or inhibiting) neuroblastoma cells. The screening method is used here with the LOPAC compound collection. (LOPAC library, Sigma). 
         [0134]    Candidate NB tumor-initiating cells (TICs) were isolated. These TICs were used in the screening assay for the identification of these kinds of compounds because they are related to SKPs (SKin-derived Precursors). For example, both SKPs and TICs originate from the neural crest, express similar neural crest markers, and differentiate in vitro into similar cell types. The availability of two neural crest stem cell sources, one from the NB tumor and the other from the skin of the same patient, affords an approach for the therapeutic target discovery provided here. 
       Materials and Methods: 
     Methods: 
       [0135]    To identify compounds that specifically target neuroblastoma TICs, a cell-based assay in which TICs from a NB patient and normal SKPs were tested in parallel. Cells were treated with test compound prior to incubation with a cell viability dye. For both cell sources, signal variability was low and the Z′ and Z factors were &gt;0.5, suggesting excellent assay quality. Hits were defined as compounds whose signals were shifted at least 3 standard deviations from the mean. 
       Results: 
       [0136]    Compounds that Affect NB Cells and Normal Cells 
         [0137]    From 3 libraries of compounds, the LOPAC collection, the Prestwick Collection and the Spectrum Collection, 46 compounds were found to affect both normal and NB cells. These 46 compounds are listed in Table 13. 
         [0000]                              TABLE 13               Normal and Neuroblastoma Hits                                    3-METHYLORSELLINIC ACID           5alpha-CHOLESTAN-3beta-OL-6-ONE           5-AZACYTIDINE           AKLAVINE HYDROCHLORIDE           Alexidine dihydrochloride           Ammonium pyrrolidinedithiocarbamate           Anisomycin           Brefeldin A from Penicillium brefeldianum           Calmidazolium chloride           Cephaeline dihydrochloride heptahydrate           CETRIMONIUM BROMIDE           CHELIDONINE (+)           COLCHICEINE           DACTINOMYCIN           Daunorubicin hydrochloride           DEOXYSAPPANONE B 7.3′-DIMETHYL           ETHER           Digitoxigenin           Digoxin           DIHYDROGAMBOGIC ACID           Dihydroouabain           Disulfiram           EMETINE           GENTIAN VIOLET           JUGLONE           LANATOSIDE C           LYCORINE           Methyl benzethonium chloride           MITOMYCIN C           Mitoxantrone           OXYPHENBUTAZONE           PATULIN           PERIPLOCYMARIN           PERUVOSIDE           PHENYLMERCURIC ACETATE           Proscillaridin A           Puromycin dihydrochloride           PYRITHIONE ZINC           PYRROMYCIN           RETUSOQUINONE           Sanguinarine           SARMENTOGENIN           Strophanthidin           Terfenadine           THIMEROSAL           Thonzonium bromide           TOMATINE                        
Table 13: 54 Identified Compounds that Affect NB Cells
 
         [0138]    Fifty-four (54) compounds selected from the LOPAC collection, Prestwick Collection and the Spectrum Collection, were found to selectively target NB cells. These 56 compounds appear in Table 14. 
         [0000]    
       
         
               
             
               
               
             
           
               
                 TABLE 14 
               
               
                   
               
               
                 Neuroblastoma Specific Hits 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 10-HYDROXYCAMTOTHECIN 
               
               
                   
                 2.3-Dimethoxy-1.4-naphthoquinone 
               
               
                   
                 4′-DEMETHYLEPIPODOPHYLLOTOXIN 
               
               
                   
                 Amodiaquin dihydrochloride dihydrate 
               
               
                   
                 AMSACRINE HYDROCHLORIDE 
               
               
                   
                 Ancitabine hydrochloride 
               
               
                   
                 ANDROGRAPHOLIDE 
               
               
                   
                 Antimycin A 
               
               
                   
                 Azaguanine-8 
               
               
                   
                 BENZALKONIUM CHLORIDE 
               
               
                   
                 Benzethonium chloride 
               
               
                   
                 BEPRIDIL HYDROCHLORIDE 
               
               
                   
                 beta-PELTATIN 
               
               
                   
                 Camptothecin (S.+) 
               
               
                   
                 CETYLPYRIDINIUM CHLORIDE 
               
               
                   
                 CGP-74514A hydrochloride 
               
               
                   
                 Chelerythrine chloride 
               
               
                   
                 CHOLESTAN-3beta.5alpha.6beta-TRIOL 
               
               
                   
                 Ciclopirox ethanolamine 
               
               
                   
                 Clofazimine 
               
               
                   
                 CONVALLATOXIN 
               
               
                   
                 CRASSIN ACETATE 
               
               
                   
                 CRINAMINE 
               
               
                   
                 Cycloheximide 
               
               
                   
                 Cytosine-1-beta-D-arabinofuranoside 
               
               
                   
                 hydrochloride 
               
               
                   
                 Dequalinium analog. C-14 linker 
               
               
                   
                 Dequalinium dichloride 
               
               
                   
                 Diphenyleneiodonium chloride 
               
               
                   
                 DL-Stearoylcarnitine chloride 
               
               
                   
                 Doxorubicin hydrochloride 
               
               
                   
                 ERYSOLIN 
               
               
                   
                 Etoposide 
               
               
                   
                 GAMBOGIC ACID 
               
               
                   
                 Idarubicin 
               
               
                   
                 IMIDACLOPRIDE 
               
               
                   
                 LIMONIN 
               
               
                   
                 Loratadine 
               
               
                   
                 MECHLORETHAMINE 
               
               
                   
                 MECLIZINE HYDROCHLORIDE 
               
               
                   
                 MG 624 
               
               
                   
                 Mycophenolic acid 
               
               
                   
                 Ouabain 
               
               
                   
                 OXYBENDAZOLE 
               
               
                   
                 Paclitaxel 
               
               
                   
                 PARAROSANILINE PAMOATE 
               
               
                   
                 Parthenolide 
               
               
                   
                 Podophyllotoxin 
               
               
                   
                 Primaquine diphosphate 
               
               
                   
                 Quinacrine dihydrochloride 
               
               
                   
                 Scoulerine 
               
               
                   
                 Taxol 
               
               
                   
                 TENIPOSIDE 
               
               
                   
                 Vinblastine sulfate salt 
               
               
                   
                 Vincristine sulfate 
               
               
                   
                   
               
             
          
         
       
     
         [0139]    Four (4) compounds selected from the LOPAC collection, Prestwick Collection and the Spectrum Collection, were found to successfully treat a NB patient and were selected as NB specific hits according to the assay criteria provided herein. These compounds serve as positive controls in the selection and screening methods. These results emphasize the validity of the assay in identifying active agents for treating neuroblastoma. These 4 compounds are listed in Table 15. 
         [0000]    
       
         
               
             
               
               
             
           
               
                 TABLE 15 
               
               
                   
               
               
                 4 Identified Compounds that are used to Treat the NB Patient 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Patient Hits (i.e. drugs that were used to treat patient AND 
               
               
                   
                 were selected as NB specific hits) 
               
               
                   
                 Ancitabine hydrochloride (aka cyclocytidine) 
               
               
                   
                 Doxorubicin hydrochloride (aka adriamycin) 
               
               
                   
                 Etoposide 
               
               
                   
                 Vincristine sulfate 
               
               
                   
                   
               
             
          
         
       
     
         [0140]    These results suggest that patient-specific therapeutics as well as novel molecular effectors of neuroblastoma can be identified using this assay. 
       Example 3 
     Cumulative Screening Assay Selection Results 
       [0141]    The present example presents the tabulated data obtained with the various chemical library screens conducted. 
         [0000]    
       
         
               
               
               
               
               
               
               
             
           
               
                 TABLE 16 
               
               
                   
               
               
                   
                   
                   
                 NB 
                   
                   
                   
               
               
                   
                   
                   
                 hit 
                 NB + FS 
                 IC50 
               
               
                 Library 
                 Name 
                 Repeated 
                 only 
                 hit 
                 test? 
                 target/mechanism 
               
               
                   
               
             
             
               
                 S 
                 
                   10-HYDROXYCAMTOTHECIN 
                 
                 X 
                 X 
                   
                   
                 modified camptothecin 
               
               
                 L 
                 2.3-Dimethoxy-1.4-naphthoquinone 
                 X 
                 X 
                   
                   
                 ROS modulator/Redox 
               
               
                   
                   
                   
                   
                   
                   
                 cycling agent used to 
               
               
                   
                   
                   
                   
                   
                   
                 study role of ROS 
               
               
                 S 
                 3-METHYLORSELLINIC ACID 
                 X 
                   
                 X 
                   
                 Aspergillus terreus 
               
               
                   
                   
                   
                   
                   
                   
                 fungal metabolite; 
               
               
                   
                   
                   
                   
                   
                   
                 possible antioxidant 
               
               
                 S 
                 4′- 
                 X 
                 X 
               
               
                   
                 DEMETHYLEPIPODOPHYLLOTOXIN 
               
               
                 S 
                 5alpha-CHOLESTAN-3beta-OL-6-ONE 
                 X 
                 X 
                   
                   
                 Cholesterol oxidation 
               
               
                   
                   
                   
                   
                   
                   
                 product; cytotoxic due to 
               
               
                   
                   
                   
                   
                   
                   
                 oxidative stress or 
               
               
                   
                   
                   
                   
                   
                   
                 cytoskeleton disruption 
               
               
                 S 
                 5-AZACYTIDINE 
                 X 
                   
                 X 
               
               
                 S 
                 
                   ACRIFLAVINIUM 
                 
                   
                   
                   
                   
                 intercalating agent that 
               
               
                   
                 
                   HYDROCHLORIDE 
                 
                   
                   
                   
                   
                 interferes with DNA 
               
               
                   
                   
                   
                   
                   
                   
                 replic/transcription; 
               
               
                   
                   
                   
                   
                   
                   
                 antitumor, 
               
               
                   
                   
                   
                   
                   
                   
                 antiproliferative 
               
               
                 S 
                 ACRISORCIN 
                   
                   
                   
                   
                 topical anti-infective 
               
               
                   
                   
                   
                   
                   
                   
                 from 1960s 
               
               
                 S 
                 AKLAVINE HYDROCHLORIDE 
                 X 
                   
                 X 
                 X 
                 natural product; anti- 
               
               
                   
                   
                   
                   
                   
                   
                 infective; related 
               
               
                   
                   
                   
                   
                   
                   
                 structures have broad 
               
               
                   
                   
                   
                   
                   
                   
                 activity against NIH 
               
               
                   
                   
                   
                   
                   
                   
                 tumor lines 
               
               
                 P 
                 
                   Alexidine dihydrochloride 
                 
                 X 
                   
                 X 
                   
                 phospholipase inh; oral 
               
               
                   
                   
                   
                   
                   
                   
                 gingivitis rinse 
               
               
                 S 
                 ALEXIDINE HYDROCHLORIDE 
               
               
                 P 
                 Alprostadil 
                   
                   
                   
                   
                 vasodilator; erectile 
               
               
                   
                   
                   
                   
                   
                   
                 dysfunction, pallative 
               
               
                   
                   
                   
                   
                   
                   
                 care for neonatal 
               
               
                   
                   
                   
                   
                   
                   
                 congenital heart defects 
               
               
                 L 
                 
                   Ammonium pyrrolidinedithiocarbamate 
                 
                 X 
                   
                 X 
                 X 
                 blocks NOS mRNA 
               
               
                   
                   
                   
                   
                   
                   
                 translation 
               
               
                 P 
                 !Amodiaquin dihydrochloride dihydrate 
                 X 
                 X 
                   
                 X 
                 antimalarial; treatment of 
               
               
                   
                   
                   
                   
                   
                   
                 CNS degeneration 
               
               
                   
                   
                   
                   
                   
                   
                 (Alzheimer, MS) 
               
               
                 S 
                 !AMODIAQUINE 
                 X 
                 X 
                   
                   
                 antimalarial; 4- 
               
               
                   
                 DIHYDROCHLORIDE 
                   
                   
                   
                   
                 aminoquinoline family; 
               
               
                   
                   
                   
                   
                   
                   
                 narrow therapeutic/toxic 
               
               
                   
                   
                   
                   
                   
                   
                 window in children 
               
               
                 S 
                 AMSACRINE HYDROCHLORIDE 
                 X 
                 X 
                   
                 X 
                 topo II inh; used in 
               
               
                   
                   
                   
                   
                   
                   
                 AML; may also be active 
               
               
                   
                   
                   
                   
                   
                   
                 vs malaria 
               
               
                 L 
                 
                   Ancitabine hydrochloride 
                 
                 X 
                 X 
                   
                   
                 cyclocytidine HCl; 
               
               
                   
                   
                   
                   
                   
                   
                 DNA-synthesis inhibitor 
               
               
                   
                   
                   
                   
                   
                   
                 (cytosine analog); 
               
               
                   
                   
                   
                   
                   
                   
                 antileukemic 
               
               
                 S 
                 
                   ANCITABINE HYDROCHLORIDE 
                 
                 X 
                 X 
                   
                 X 
               
               
                 S 
                 
                   ANDROGRAPHOLIDE 
                 
                 X 
                 X 
                   
                 X 
                 Chinese herbal medicine; 
               
               
                   
                   
                   
                   
                   
                   
                 anti-inflamm; immune 
               
               
                   
                   
                   
                   
                   
                   
                 boosting?; anti-cancer vs 
               
               
                   
                   
                   
                   
                   
                   
                 HL60, MCF7, others 
               
               
                   
                   
                   
                   
                   
                   
                 through G0/G1 block 
               
               
                   
                   
                   
                   
                   
                   
                 and apoptosis induction 
               
               
                 P 
                 
                   Anisomycin 
                 
                 X 
                   
                 X 
                   
                 protein synthesis inh thru 
               
               
                   
                   
                   
                   
                   
                   
                 peptidyl transferase of 
               
               
                   
                   
                   
                   
                   
                   
                 80S ribosome; treatment 
               
               
                   
                   
                   
                   
                   
                   
                 activates p54, MAPK, 
               
               
                   
                   
                   
                   
                   
                   
                 SAPK 
               
               
                 P 
                 #Antimycin A 
                 X 
                 X 
                   
                 X 
                 antifungal, antimicrobial; 
               
               
                   
                   
                   
                   
                   
                   
                 blocks e-transport 
               
               
                   
                   
                   
                   
                   
                   
                 between cytochrome B 
               
               
                   
                   
                   
                   
                   
                   
                 and cytochrome C; bind 
               
               
                   
                   
                   
                   
                   
                   
                 the BH3 domain of Bcl- 
               
               
                   
                   
                   
                   
                   
                   
                 xL and induce apoptosis 
               
               
                   
                   
                   
                   
                   
                   
                 in cells overexpressing 
               
               
                   
                   
                   
                   
                   
                   
                 Bcl-2 and Bcl-xL 
               
               
                 P 
                 !Avermectin B1 
                   
                   
                   
                 X 
                 antiworm/insecticide 
               
               
                 P 
                 
                   Azaguanine-8 
                 
                 X 
                 X 
                   
                 X 
                 purine analog 
               
               
                 S 
                 BENZALKONIUM CHLORIDE 
                 X 
                 X 
                   
                   
                 cationic detergent; v 
               
               
                   
                   
                   
                   
                   
                   
                 common antiseptic and 
               
               
                   
                   
                   
                   
                   
                   
                 preservative 
               
               
                 P 
                 +Benzethonium chloride 
                 X 
                 X 
                   
                   
                 topical antimicrobial 
               
               
                   
                   
                   
                   
                   
                   
                 used in cosmetics as 
               
               
                   
                   
                   
                   
                   
                   
                 preservative 
               
               
                 S 
                 +BENZALKONIUM CHLORIDE 
                 X 
                 X 
               
               
                 S 
                 !BEPRIDIL HYDROCHLORIDE 
                 X 
                 X 
                   
                 X 
                 nonselective Ca channel 
               
               
                   
                   
                   
                   
                   
                   
                 blocker used for 
               
               
                   
                   
                   
                   
                   
                   
                 treatment of chronic 
               
               
                   
                   
                   
                   
                   
                   
                 angina pectoris; alters 
               
               
                   
                   
                   
                   
                   
                   
                 potential dep and 
               
               
                   
                   
                   
                   
                   
                   
                 receptor-operated Ca 
               
               
                   
                   
                   
                   
                   
                   
                 channels and inhibits fast 
               
               
                   
                   
                   
                   
                   
                   
                 Na inward currents 
               
               
                 S 
                 beta-PELTATIN 
                 X 
                 X 
                   
                 X 
                 extracted from Mayapple 
               
               
                   
                   
                   
                   
                   
                   
                 rhizome (like 
               
               
                   
                   
                   
                   
                   
                   
                 podophyllotoxin); some 
               
               
                   
                   
                   
                   
                   
                   
                 evidence of in vitro anti- 
               
               
                   
                   
                   
                   
                   
                   
                 tumor f/x but vague 
               
               
                 L 
                 Brefeldin A from Penicillium 
                 X 
                   
                 X 
                   
                 fungal metabolite that 
               
               
                   
                 brefeldianum 
                   
                   
                   
                   
                 disrupts Golgi structure 
               
               
                   
                   
                   
                   
                   
                   
                 and function 
               
               
                 L 
                 
                   Calmidazolium chloride 
                 
                 X 
                   
                 X 
                   
                 Potent inhibitor of 
               
               
                   
                   
                   
                   
                   
                   
                 calmodulin activation of 
               
               
                   
                   
                   
                   
                   
                   
                 phosphodiesterase; 
               
               
                   
                   
                   
                   
                   
                   
                 strongly inhibits 
               
               
                   
                   
                   
                   
                   
                   
                 calmodulin-dependent 
               
               
                   
                   
                   
                   
                   
                   
                 Ca2+-ATPase 
               
               
                 S 
                 CAMPTOTHECIN 
                 X 
                 X 
               
               
                 L 
                 
                   Camptothecin (S.+) 
                 
                 X 
                 X 
                   
                   
                 topo 1 inh 
               
               
                 P 
                 
                   Camptothecine (S.+) 
                 
                 X 
                 X 
                   
                 X 
               
               
                 P 
                 Cephaeline dihydrochloride heptahydrate 
                 X 
                   
                 X 
                   
                 ipecac alkaloid 
               
               
                 S 
                 CETRIMONIUM BROMIDE 
                 X 
                   
                 X 
                   
                 cationic detergent; 
               
               
                   
                   
                   
                   
                   
                   
                 quaternary ammonium 
               
               
                   
                   
                   
                   
                   
                   
                 compound used in hair 
               
               
                   
                   
                   
                   
                   
                   
                 conditioner and as a 
               
               
                   
                   
                   
                   
                   
                   
                 antimicrobial; tested as a 
               
               
                   
                   
                   
                   
                   
                   
                 lavage during colon 
               
               
                   
                   
                   
                   
                   
                   
                 resections...no benefit 
               
               
                   
                   
                   
                   
                   
                   
                 and potentially toxic 
               
               
                 S 
                 CETYLPYRIDINIUM CHLORIDE 
                 X 
                 X 
                   
                   
                 active ingredient in 
               
               
                   
                   
                   
                   
                   
                   
                 Scope; antiseptic used in 
               
               
                   
                   
                   
                   
                   
                   
                 oral rinses 
               
               
                 L 
                 
                   CGP-74514A hydrochloride 
                 
                 X 
                 X 
                   
                 X 
                 Cdk1 inh 
               
               
                 L 
                 
                   Chelerythrine chloride 
                 
                 X 
                 X 
                   
                 X 
                 PKC inhibitor; affects 
               
               
                   
                   
                   
                   
                   
                   
                 translocation of PKC 
               
               
                   
                   
                   
                   
                   
                   
                 from cytosol to plasma 
               
               
                   
                   
                   
                   
                   
                   
                 membrane 
               
               
                 S 
                 
                   CHELIDONINE (+) 
                 
                 X 
                 X 
                   
                   
                 G2/M arrest associated 
               
               
                   
                   
                   
                   
                   
                   
                 with increased cycB1 
               
               
                   
                   
                   
                   
                   
                   
                 levels, cdc2 activity and 
               
               
                   
                   
                   
                   
                   
                   
                 SAPK/JNK activity; 
               
               
                   
                   
                   
                   
                   
                   
                 weak tubulin interaction; 
               
               
                   
                   
                   
                   
                   
                   
                 induced apoptosis at 
               
               
                   
                   
                   
                   
                   
                   
                 1uM in Jurkat cells 
               
               
                 S 
                 CHOLESTAN-3beta.5alpha.6beta- 
                 X 
                 X 
                   
                 X 
                 Cholesterol oxidation 
               
               
                   
                 TRIOL 
                   
                   
                   
                   
                 product; cytotoxic due to 
               
               
                   
                   
                   
                   
                   
                   
                 oxidative stress or 
               
               
                   
                   
                   
                   
                   
                   
                 cytoskeleton disruption 
               
               
                 P 
                 Ciclopirox ethanolamine 
                 X 
                 X 
                   
                   
                 topical antifungal, anti- 
               
               
                   
                   
                   
                   
                   
                   
                 inflammatory 
               
               
                 S 
                 CICLOPIROX OLAMINE 
                 X 
                 X 
                   
                 X 
               
               
                 P 
                 Clofazimine 
                 X 
                 X 
                   
                 X 
                 leprosy treatment; anti- 
               
               
                   
                   
                   
                   
                   
                   
                 inflammatory f/x; 
               
               
                   
                   
                   
                   
                   
                   
                 disrupts cc by binding 
               
               
                   
                   
                   
                   
                   
                   
                 DNA, may bind K+ 
               
               
                   
                   
                   
                   
                   
                   
                 transporters 
               
               
                 S 
                 
                   COLCHICEINE 
                 
                 X 
                   
                 X 
                   
                 metabolite of colchicine; 
               
               
                   
                   
                   
                   
                   
                   
                 less toxic to hepatocytes; 
               
               
                   
                   
                   
                   
                   
                   
                 less binding to tubulin 
               
               
                   
                   
                   
                   
                   
                   
                 but presumably has 
               
               
                   
                   
                   
                   
                   
                   
                 similar modeof action 
               
               
                 L 
                 Colchicine 
                 X 
                 X 
                   
                   
                 binds tubulin/blocks 
               
               
                   
                   
                   
                   
                   
                   
                 mitosis by preventing 
               
               
                   
                   
                   
                   
                   
                   
                 spindle formation; 
               
               
                   
                   
                   
                   
                   
                   
                 bioactive doses would be 
               
               
                   
                   
                   
                   
                   
                   
                 toxic 
               
               
                 P 
                 Colchicine 
                 X 
                 X 
                   
                 X 
               
               
                 S 
                 
                   COLCHICINE 
                 
               
               
                 S 
                 #CONVALLATOXIN 
                 X 
                 X 
                   
                 X 
                 derived from lily of the 
               
               
                   
                   
                   
                   
                   
                   
                 valley; digitalis-like 
               
               
                   
                   
                   
                   
                   
                   
                 action 
               
               
                 S 
                 CRASSIN ACETATE 
                 X 
                 X 
                   
                 X 
                 antineoplastic vs P388 
               
               
                   
                   
                   
                   
                   
                   
                 leukemia and HT29 
               
               
                   
                   
                   
                   
                   
                   
                 colon cancer cells in 
               
               
                   
                   
                   
                   
                   
                   
                 vitro; extracted from 
               
               
                   
                   
                   
                   
                   
                   
                 marine invertebrates 
               
               
                 S 
                 CRINAMINE 
                 X 
                 X 
                   
                 X 
               
               
                 P 
                 
                   Cycloheximide 
                 
                 X 
                 X 
                   
                   
                 protein synthesis inh 
               
               
                 S 
                 
                   CYCLOHEXIMIDE 
                 
               
               
                 S 
                 CYMARIN 
                   
                   
                 X 
               
               
                 S 
                 
                   CYTARABINE 
                 
                 X 
                   
                   
                   
                 Ara-C; DNA damage, S- 
               
               
                   
                   
                   
                   
                   
                   
                 phase block; inh DNA/ 
               
               
                   
                   
                   
                   
                   
                   
                 RNA pol 
               
               
                 L 
                 
                   Cytosine-1-beta-D-arabinofuranoside 
                 
                 X 
                 X 
                   
                   
                 Ara-C; selective inh of 
               
               
                   
                 
                   hydrochloride 
                 
                   
                   
                   
                   
                 DNA synthesis 
               
               
                 S 
                 DACTINOMYCIN 
                 X 
                   
                 X 
               
               
                 P 
                 
                   Daunorubicin hydrochloride 
                 
                 X 
                   
                 X 
                   
                 DNA intercalator; 
               
               
                   
                   
                   
                   
                   
                   
                 neuroblastoma treatment 
               
               
                 S 
                 DEOXYSAPPANONE B 7.3′- 
                 X 
                   
                 X 
                   
                 flavanoid derived from 
               
               
                   
                 DIMETHYL ETHER 
                   
                   
                   
                   
                 Caesalpinia sappan tree; 
               
               
                   
                   
                   
                   
                   
                   
                 Chinese med treatment 
               
               
                   
                   
                   
                   
                   
                   
                 for tumor, diarrhea; 
               
               
                   
                   
                   
                   
                   
                   
                 aldose reductase 
               
               
                   
                   
                   
                   
                   
                   
                 inhibitor?; one study 
               
               
                   
                   
                   
                   
                   
                   
                 suggesting activity vs 
               
               
                   
                   
                   
                   
                   
                   
                 head and neck cancer 
               
               
                   
                   
                   
                   
                   
                   
                 cell line 
               
               
                 L 
                 
                   Dequalinium analog. C-14 linker 
                 
                 X 
                 X 
                   
                 X 
                 Protein kinase C-alpha 
               
               
                   
                   
                   
                   
                   
                   
                 (PKC-alpha) inhibitor 
               
               
                 P 
                 !Dequalinium dichloride 
                 X 
                 X 
                   
                 X 
                 Selective blocker of 
               
               
                   
                   
                   
                   
                   
                   
                 apamin-sensitive K+ 
               
               
                   
                   
                   
                   
                   
                   
                 channels 
               
               
                 L 
                 !Dequalinium dichloride 
                 X 
                 X 
                   
                 X 
                 Member of delocalized 
               
               
                   
                   
                   
                   
                   
                   
                 lipophilic cations 
               
               
                   
                   
                   
                   
                   
                   
                 (DLCs), a family of 
               
               
                   
                   
                   
                   
                   
                   
                 compounds that 
               
               
                   
                   
                   
                   
                   
                   
                 accumulate in 
               
               
                   
                   
                   
                   
                   
                   
                 mitochondria driven by 
               
               
                   
                   
                   
                   
                   
                   
                 the negative 
               
               
                   
                   
                   
                   
                   
                   
                 transmembrane potential; 
               
               
                   
                   
                   
                   
                   
                   
                 inhibitor of NADH- 
               
               
                   
                   
                   
                   
                   
                   
                 ubiquinone reductase; A 
               
               
                   
                   
                   
                   
                   
                   
                 novel mitochondria 
               
               
                   
                   
                   
                   
                   
                   
                 delivery system is based 
               
               
                   
                   
                   
                   
                   
                   
                 on dequalinium. This 
               
               
                   
                   
                   
                   
                   
                   
                 DLC forms liposome- 
               
               
                   
                   
                   
                   
                   
                   
                 like aggregates termed 
               
               
                   
                   
                   
                   
                   
                   
                 ‘DQAsomes’. DQAsomes 
               
               
                   
                   
                   
                   
                   
                   
                 are being tested as 
               
               
                   
                   
                   
                   
                   
                   
                 mitochondria drug 
               
               
                   
                   
                   
                   
                   
                   
                 delivery systems for 
               
               
                   
                   
                   
                   
                   
                   
                 small molecules such as 
               
               
                   
                   
                   
                   
                   
                   
                 paclitaxel 
               
               
                 P 
                 #Digitoxigenin 
                 X 
                   
                 X 
                   
                 Digitalis derivative; 
               
               
                   
                   
                   
                   
                   
                   
                 blocks Na+/K+ pump 
               
               
                 S 
                 #DIGITOXIN 
               
               
                 P 
                 #Digoxigenin 
                 X 
                 X 
                   
                   
                 Digitalis derivative; 
               
               
                   
                   
                   
                   
                   
                   
                 blocks Na+/K+ pump 
               
               
                 P 
                 #Digoxin 
                 X 
                   
                 X 
                 X 
                 Digitalis derivative; 
               
               
                   
                   
                   
                   
                   
                   
                 blocks Na+/K+ pump 
               
               
                 S 
                 #DIGOXIN 
                 X 
                   
                 X 
               
               
                 S 
                 DIHYDROGAMBOGIC ACID 
                 X 
                   
                 X 
                 X 
               
               
                 L 
                 !Dihydroouabain 
                 X 
                   
                 X 
                 X 
                 Na+/K+ pump inhibitor 
               
               
                 L 
                 
                   Diphenyleneiodonium chloride 
                 
                 X 
                 X 
                   
                 X 
                 eNOS inh (endothelial 
               
               
                   
                   
                   
                   
                   
                   
                 NOS) 
               
               
                 S 
                 DISULFIRAM 
                 X 
                 X 
               
               
                 P 
                 Disulfiram 
                 X 
                   
                 X 
                   
                 antabuse, rxn with 
               
               
                   
                   
                   
                   
                   
                   
                 alcohol use 
               
               
                 L 
                 
                   DL-Stearoylcarnitine chloride 
                 
                 X 
                 X 
                   
                 X 
                 PKC inh 
               
               
                 P 
                 
                   Doxorubicin hydrochloride 
                 
                 X 
                 X 
                   
                   
                 DNA synthesis inh; 
               
               
                   
                   
                   
                   
                   
                   
                 stabilizes topo II 
               
               
                   
                   
                   
                   
                   
                   
                 complex after strand 
               
               
                   
                   
                   
                   
                   
                   
                 cleavage 
               
               
                 S 
                 
                   EMETINE 
                 
                 X 
                   
                 X 
               
               
                 P 
                 
                   Emetine dihydrochloride 
                 
                 X 
                   
                 X 
                   
                 ipecac alkaloid; inh 
               
               
                   
                   
                   
                   
                   
                   
                 protein synthesis by 
               
               
                   
                   
                   
                   
                   
                   
                 blocking Rb movement 
               
               
                   
                   
                   
                   
                   
                   
                 on mRNA; inhibit DNA 
               
               
                   
                   
                   
                   
                   
                   
                 replication in S phase 
               
               
                 L 
                 
                   Emetine dihydrochloride hydrate 
                 
                 X 
                   
                 X 
                   
                 Apoptosis inducer; 
               
               
                   
                   
                   
                   
                   
                   
                 RNA-Protein translation 
               
               
                   
                   
                   
                   
                   
                   
                 inhibitor 
               
               
                 S 
                 ERYSOLIN 
                 X 
                 X 
                   
                 X 
                 organic isothiocyanate 
               
               
                   
                   
                   
                   
                   
                   
                 found in cruciferous 
               
               
                   
                   
                   
                   
                   
                   
                 veggies; increases 
               
               
                   
                   
                   
                   
                   
                   
                 accumulation of chemo 
               
               
                   
                   
                   
                   
                   
                   
                 drugs in PANC-1, MCF- 
               
               
                   
                   
                   
                   
                   
                   
                 7, NCI-H460 cell lines 
               
               
                 P 
                 
                   Etoposide 
                 
                 X 
                 X 
                   
                 X 
                 topo II inh 
               
               
                 L 
                 
                   Etoposide 
                 
                 X 
                 X 
               
               
                 P 
                 Fosfosal 
                   
                   
                   
                   
                 salicylic acid derivative/ 
               
               
                   
                   
                   
                   
                   
                   
                 anti-inflammatory 
               
               
                 S 
                 
                   GAMBOGIC ACID 
                 
                 X 
                 X 
                   
                 X 
                 principle pigment of 
               
               
                   
                   
                   
                   
                   
                   
                 gambage resin (bright 
               
               
                   
                   
                   
                   
                   
                   
                 orange); caspase 
               
               
                   
                   
                   
                   
                   
                   
                 activator (not well 
               
               
                   
                   
                   
                   
                   
                   
                 characterized); growth/ 
               
               
                   
                   
                   
                   
                   
                   
                 tumor inhibitory vs 
               
               
                   
                   
                   
                   
                   
                   
                 HeLa, HEL, gastic 
               
               
                   
                   
                   
                   
                   
                   
                 cancer, lung carcinoma 
               
               
                   
                   
                   
                   
                   
                   
                 cell lines 
               
               
                 S 
                 GENTIAN VIOLET 
                 X 
                   
                 X 
               
               
                 L 
                 
                   Idarubicin 
                 
                 X 
                 X 
                   
                 X 
                 antineoplastic, DNA 
               
               
                   
                   
                   
                   
                   
                   
                 metabolism 
               
               
                 S 
                 !IMIDACLOPRIDE 
                 X 
                 X 
                   
                 X 
                 a4b2 nAChR agonist; 
               
               
                   
                   
                   
                   
                   
                   
                 activates ERK pathway; 
               
               
                   
                   
                   
                   
                   
                   
                 insecticide 
               
               
                 S 
                 JUGLONE 
                 X 
                   
                 X 
                 X 
                 Pin1 inh; alkylates 
               
               
                   
                   
                   
                   
                   
                   
                 thioredoxin reductase; 
               
               
                   
                   
                   
                   
                   
                   
                 PI3K inh?; inhibits 
               
               
                   
                   
                   
                   
                   
                   
                 growth of HCT-15, 
               
               
                   
                   
                   
                   
                   
                   
                 HeLa, HL60 cell lines 
               
               
                 P 
                 Kaempferol 
                   
                   
                   
                   
                 antioxidant/flavenoid 
               
               
                 P 
                 #Lanatoside C 
                 X 
                   
                 X 
                   
                 Digitalis derivative; 
               
               
                   
                   
                   
                   
                   
                   
                 blocks Na+/K+ pump 
               
               
                 S 
                 #LANATOSIDE C 
                 X 
                   
                 X 
               
               
                 S 
                 LIMONIN 
                 X 
                 X 
                   
                 X 
                 isolated from citrus fruit 
               
               
                   
                   
                   
                   
                   
                   
                 seeds; inhibits HIV1 
               
               
                   
                   
                   
                   
                   
                   
                 protease activity; 
               
               
                   
                   
                   
                   
                   
                   
                 antinociceptive, inhibits 
               
               
                   
                   
                   
                   
                   
                   
                 MCF7 growth but not 
               
               
                   
                   
                   
                   
                   
                   
                 other cancer cell lines 
               
               
                 L 
                 *Loratadine 
                 X 
                 X 
                   
                 X 
                 H1 Histamine R 
               
               
                   
                   
                   
                   
                   
                   
                 antagonist 
               
               
                 S 
                 LYCORINE 
                 X 
                   
                 X 
               
               
                 P 
                 +Mebendazole 
                   
                   
                   
                   
                 anthelmintic; blocks 
               
               
                   
                   
                   
                   
                   
                   
                 glucose/nutrient uptake 
               
               
                   
                   
                   
                   
                   
                   
                 in adult worm intestine; 
               
               
                   
                   
                   
                   
                   
                   
                 reported to be a mitotic 
               
               
                   
                   
                   
                   
                   
                   
                 spindle poison (resulting 
               
               
                   
                   
                   
                   
                   
                   
                 in chromosomal 
               
               
                   
                   
                   
                   
                   
                   
                 nondisjunction) 
               
               
                 S 
                 
                   MECHLORETHAMINE 
                 
                 X 
                 X 
                   
                 X 
                 mustard gas derivative; 
               
               
                   
                   
                   
                   
                   
                   
                 polyfunctional alkylating 
               
               
                   
                   
                   
                   
                   
                   
                 agent = DNA breaks and 
               
               
                   
                   
                   
                   
                   
                   
                 crosslinks; non cc phase 
               
               
                   
                   
                   
                   
                   
                   
                 specific 
               
               
                 S 
                 *MECLIZINE HYDROCHLORIDE 
                 X 
                 X 
                   
                 X 
                 antivert/bonine; motion 
               
               
                   
                   
                   
                   
                   
                   
                 sickness/vertigo 
               
               
                   
                   
                   
                   
                   
                   
                 treatment; piperazine 
               
               
                   
                   
                   
                   
                   
                   
                 class of antihistamines 
               
               
                 L 
                 
                   Melphalan 
                 
                   
                   
                   
                   
                 Antineoplastic; forms 
               
               
                   
                   
                   
                   
                   
                   
                 DNA intrastrand 
               
               
                   
                   
                   
                   
                   
                   
                 crosslinks by 
               
               
                   
                   
                   
                   
                   
                   
                 bifunctional alkylation in 
               
               
                   
                   
                   
                   
                   
                   
                 5′-GGC sequences; used 
               
               
                   
                   
                   
                   
                   
                   
                 in NB megatherapy 
               
               
                 P 
                 Menadione 
                   
                   
                   
                   
                 vitamin K3 (vitK2 
               
               
                   
                   
                   
                   
                   
                   
                 precursor); reacts with — 
               
               
                   
                   
                   
                   
                   
                   
                 SH/soaks up GSH = high 
               
               
                   
                   
                   
                   
                   
                   
                 ROS = altered Ca2+ = Ca- 
               
               
                   
                   
                   
                   
                   
                   
                 dep DNA fragmentation; 
               
               
                   
                   
                   
                   
                   
                   
                 toxic at high doses so 
               
               
                   
                   
                   
                   
                   
                   
                 vitK2 currently being 
               
               
                   
                   
                   
                   
                   
                   
                 used in cancer trials 
               
               
                 P 
                 +Methiazole 
                   
                   
                   
                   
                 anthelmintic 
               
               
                 P 
                 +Methyl benzethonium 
                 X 
                   
                 X 
                   
                 topical antimicrobial 
               
               
                 S 
                 +METHYLBENZETHONIUM 
                 X 
                   
                 X 
               
               
                   
                 CHLORIDE 
               
               
                 L 
                 !MG 624 
                 X 
                 X 
                   
                 X 
                 Nicotinic acetylcholine 
               
               
                   
                   
                   
                   
                   
                   
                 receptor antagonist; 
               
               
                   
                   
                   
                   
                   
                   
                 selectively inhibits 
               
               
                   
                   
                   
                   
                   
                   
                 alpha-bungarotoxin 
               
               
                   
                   
                   
                   
                   
                   
                 sensitive receptors that 
               
               
                   
                   
                   
                   
                   
                   
                 contain the alpha7 
               
               
                   
                   
                   
                   
                   
                   
                 subunit 
               
               
                 S 
                 MITOMYCIN C 
                 X 
                   
                 X 
                 X 
               
               
                 S 
                 
                   MITOXANTHRONE 
                 
                   
                   
                   
                 X 
               
               
                   
                 
                   HYDROCHLORIDE 
                 
               
               
                 L 
                 
                   Mitoxantrone 
                 
                 X 
                   
                 X 
                   
                 topo II inh; used in ALL, 
               
               
                   
                   
                   
                   
                   
                   
                 breast cancer, non- 
               
               
                   
                   
                   
                   
                   
                   
                 hodgkin&#39;s lymphoma 
               
               
                 P 
                 
                   Mitoxantrone dihydrochloride 
                 
                 X 
                   
                 X 
               
               
                 P 
                 
                   Mycophenolic acid 
                 
                 X 
                 X 
                   
                 X 
                 immunosuppressant; 
               
               
                   
                   
                   
                   
                   
                   
                 blocks de novo purine 
               
               
                   
                   
                   
                   
                   
                   
                 biosynthesis 
               
               
                 S 
                 NERIIFOLIN 
               
               
                 L 
                 #Ouabain 
                 X 
                   
                 X 
                   
                 Blocks movement of the 
               
               
                   
                   
                   
                   
                   
                   
                 H5 and H6 
               
               
                   
                   
                   
                   
                   
                   
                 transmembrane domains 
               
               
                   
                   
                   
                   
                   
                   
                 of Na+-K+ ATPases 
               
               
                 S 
                 #OUABAIN 
                 X 
                   
                 X 
                 X 
               
               
                 S 
                 +OXYBENDAZOLE 
                 X 
                 X 
                   
                 X 
                 benzimidazole 
               
               
                   
                   
                   
                   
                   
                   
                 anthelmintic used in 
               
               
                   
                   
                   
                   
                   
                   
                 horses and other 
               
               
                   
                   
                   
                   
                   
                   
                 ruminants 
               
               
                 S 
                 OXYPHENBUTAZONE 
                 X 
                   
                 X 
                   
                 Anti-inflammatory 
               
               
                   
                   
                   
                   
                   
                   
                 (Tandearil); binds 
               
               
                   
                   
                   
                   
                   
                   
                 phospholipase A2, 
               
               
                   
                   
                   
                   
                   
                   
                 human neutrophil 
               
               
                   
                   
                   
                   
                   
                   
                 elastase 
               
               
                 P 
                 
                   Paclitaxel 
                 
                 X 
                 X 
                   
                 X 
                 taxol 
               
               
                 S 
                 
                   PACLITAXEL 
                 
                 X 
                 X 
                   
                   
                 taxol 
               
               
                 S 
                 PARAROSANILINE PAMOATE 
                 X 
                   
                 X 
                 X 
               
               
                 P 
                 
                   Parthenolide 
                 
                 X 
                 X 
                   
                 X 
                 feverfew extract; NFkB 
               
               
                   
                   
                   
                   
                   
                   
                 inh, p53 activ, increased 
               
               
                   
                   
                   
                   
                   
                   
                 ROS, JNK activ (indep 
               
               
                   
                   
                   
                   
                   
                   
                 of NFkB and ROS), inh 
               
               
                   
                   
                   
                   
                   
                   
                 of MAPK/ERK pathway 
               
               
                 S 
                 
                   PARTHENOLIDE 
                 
                 X 
                 X 
                   
                   
                 seems to work best as a 
               
               
                   
                   
                   
                   
                   
                   
                 chemosensitizer...studies 
               
               
                   
                   
                   
                   
                   
                   
                 in breast, skin, 
               
               
                   
                   
                   
                   
                   
                   
                 pancreatic, thoracic cell 
               
               
                   
                   
                   
                   
                   
                   
                 lines 
               
               
                 S 
                 PATULIN 
                 X 
                   
                 X 
               
               
                 S 
                 #PERIPLOCYMARIN 
                 X 
                   
                 X 
                 X 
                 digoxin relative 
               
               
                 S 
                 #PERUVOSIDE 
                 X 
                   
                 X 
                 X 
                 inhibitor of Na+K+- 
               
               
                   
                   
                   
                   
                   
                   
                 ATPase; cardiac 
               
               
                   
                   
                   
                   
                   
                   
                 glycoside class 
               
               
                 S 
                 PHENYLMERCURIC ACETATE 
                 X 
                   
                 X 
               
               
                 P 
                 
                   Podophyllotoxin 
                 
                 X 
                 X 
                   
                 X 
                 etoposide precursor/ 
               
               
                   
                   
                   
                   
                   
                   
                 Antineoplastic glucoside; 
               
               
                   
                   
                   
                   
                   
                   
                 inhibitor of microtubule 
               
               
                   
                   
                   
                   
                   
                   
                 assembly; G2/M cc 
               
               
                   
                   
                   
                   
                   
                   
                 arrest 
               
               
                 L 
                 
                   Podophyllotoxin 
                 
                 X 
                 X 
               
               
                 S 
                 
                   PODOPHYLLOTOXIN ACETATE 
                 
                 X 
                   
                 X 
               
               
                 P 
                 !Primaquine diphosphate 
                 X 
                 X 
                   
                 X 
                 antimalarial/inh of 
               
               
                   
                   
                   
                   
                   
                   
                 DNA, RNA, protein 
               
               
                   
                   
                   
                   
                   
                   
                 synthesis/muscarinic 
               
               
                   
                   
                   
                   
                   
                   
                 AChR inh 
               
               
                 P 
                 #Proscillaridin A 
                 X 
                   
                 X 
                   
                 Na+/K+ ATPase inh; 
               
               
                   
                   
                   
                   
                   
                   
                 digitalis related 
               
               
                 P 
                 
                   Puromycin dihydrochloride 
                 
                 X 
                   
                 X 
                   
                 protein synthesis inh, 
               
               
                   
                   
                   
                   
                   
                   
                 premature strand 
               
               
                   
                   
                   
                   
                   
                   
                 termination 
               
               
                 S 
                 
                   PUROMYCIN HYDROCHLORIDE 
                 
                 X 
                   
                 X 
               
               
                 S 
                 PYRITHIONE ZINC 
                 X 
                   
                 X 
                 X 
               
               
                 S 
                 PYRROMYCIN 
                 X 
                   
                 X 
                   
                 anthracycline derivative; 
               
               
                   
                   
                   
                   
                   
                   
                 monosaccharide; induces 
               
               
                   
                   
                   
                   
                   
                   
                 erythroid diff in K562 
               
               
                 P 
                 +Pyrvinium pamoate 
                   
                   
                   
                   
                 pinworm treatment; 
               
               
                   
                   
                   
                   
                   
                   
                 prevents gluccose 
               
               
                   
                   
                   
                   
                   
                   
                 uptake; antitumor 
               
               
                   
                   
                   
                   
                   
                   
                 activity vs pancreatic cell 
               
               
                   
                   
                   
                   
                   
                   
                 line in SCID model, see 
               
               
                   
                   
                   
                   
                   
                   
                 decrease Akt phos 
               
               
                 L 
                 !Quinacrine dihydrochloride 
                 X 
                 X 
                   
                 X 
                 Monoamine oxidase 
               
               
                   
                   
                   
                   
                   
                   
                 (MAO) inhibitor; 
               
               
                   
                   
                   
                   
                   
                   
                 antimalarial 
               
               
                 P 
                 !Quinacrine dihydrochloride dihydrate 
                 X 
                 X 
                   
                   
                 Antimalarial, causes 
               
               
                   
                   
                   
                   
                   
                   
                 female sterility 
               
               
                 S 
                 RETUSOQUINONE 
                 X 
                   
                 X 
                   
                 ? 
               
               
                 P 
                 !Sanguinarine 
                 X 
                   
                 X 
                   
                 Inhibitor of Mg2+ and 
               
               
                   
                   
                   
                   
                   
                   
                 Na+/K+-ATPase; 
               
               
                   
                   
                   
                   
                   
                   
                 isolated from the leaves 
               
               
                   
                   
                   
                   
                   
                   
                 and stems of Macleaya 
               
               
                   
                   
                   
                   
                   
                   
                 cordata and microcarpa 
               
               
                 L 
                 !Sanguinarine chloride 
                 X 
                   
                 X 
                 X 
               
               
                 S 
                 !SANGUINARINE SULFATE 
                 X 
                 X 
               
               
                 S 
                 SARMENTOGENIN 
                 X 
                   
                 X 
               
               
                 P 
                 !Scoulerine 
                 X 
                 X 
                   
                 X 
                 opium intermediate/ 
               
               
                   
                   
                   
                   
                   
                   
                 alkaloid; a1- 
               
               
                   
                   
                   
                   
                   
                   
                 adrenoreceptor inh (G- 
               
               
                   
                   
                   
                   
                   
                   
                 protein coupled R found 
               
               
                   
                   
                   
                   
                   
                   
                 on PNS sympathetic 
               
               
                   
                   
                   
                   
                   
                   
                 nerve terminals, CNS 
               
               
                   
                   
                   
                   
                   
                   
                 postsynaptically; target 
               
               
                   
                   
                   
                   
                   
                   
                 of catecholamines) 
               
               
                 P 
                 !Strophanthidin 
                 X 
                   
                 X 
                 X 
                 blocks Na+/K+ ATPase 
               
               
                   
                   
                   
                   
                   
                   
                 at high conc; opposite f/x 
               
               
                   
                   
                   
                   
                   
                   
                 at low dose (Quabain) 
               
               
                 S 
                 !STROPHANTHIDIN 
                 X 
                   
                 X 
               
               
                 S 
                 !STROPHANTHIDINIC ACID 
                 X 
                 X 
               
               
                   
                 !LACTONE ACETATE 
               
               
                 P 
                 !Strophantine octahydrate 
                 X 
                   
                 X 
               
               
                 L 
                 
                   Taxol 
                 
                 X 
                 X 
                   
                   
                 Antitumor agent; 
               
               
                   
                   
                   
                   
                   
                   
                 promotes assembly of 
               
               
                   
                   
                   
                   
                   
                   
                 microtubules and inhibits 
               
               
                   
                   
                   
                   
                   
                   
                 tubulin disassembly 
               
               
                   
                   
                   
                   
                   
                   
                 process 
               
               
                 S 
                 
                   TENIPOSIDE 
                 
                 X 
                 X 
                   
                 X 
                 common NB treatment; 
               
               
                   
                   
                   
                   
                   
                   
                 semisynthetic 
               
               
                   
                   
                   
                   
                   
                   
                 podophyllotoxin 
               
               
                   
                   
                   
                   
                   
                   
                 derivative related to 
               
               
                   
                   
                   
                   
                   
                   
                 etoposide; topo II inh; 
               
               
                   
                   
                   
                   
                   
                   
                 induced single strand 
               
               
                   
                   
                   
                   
                   
                   
                 DNA breaks; activity in 
               
               
                   
                   
                   
                   
                   
                   
                 late S and G2 phases 
               
               
                 P 
                 *Terfenadine 
                 X 
                   
                 X 
                 X 
                 nonsedating 
               
               
                   
                   
                   
                   
                   
                   
                 antihistimineoff market 
               
               
                   
                   
                   
                   
                   
                   
                 due to cardiac f/x 
               
               
                 S 
                 THIMEROSAL 
                 X 
                   
                 X 
               
               
                 S 
                 THIRAM 
               
               
                 P 
                 Thonzonium bromide 
                 X 
                   
                 X 
                   
                 cationic detergent 
               
               
                 S 
                 TOMATINE 
                 X 
                   
                 X 
                 X 
               
               
                 P 
                 Verteporfin 
                   
                   
                   
                   
                 photoreactive dye used 
               
               
                   
                   
                   
                   
                   
                   
                 in treatment of macular 
               
               
                   
                   
                   
                   
                   
                   
                 generation; anti- 
               
               
                   
                   
                   
                   
                   
                   
                 angiogenic 
               
               
                 S 
                 
                   VINBLASTINE SULFATE 
                 
                 X 
                 X 
               
               
                 L 
                 
                   Vinblastine sulfate salt 
                 
                 X 
                 X 
                   
                 X 
                 Inhibitor of microtubule 
               
               
                   
                   
                   
                   
                   
                   
                 assembly 
               
               
                 L 
                 
                   Vincristine sulfate 
                 
                 X 
                 X 
                   
                 X 
                 Inhibitor of microtubule 
               
               
                   
                   
                   
                   
                   
                   
                 assembly 
               
               
                   
                   
                 132/151 
               
               
                   
                   
                 repeated 
               
               
                   
                   
                 (87%) 
               
               
                   
               
               
                 BOLD: DNA damage/protein synthesis inhibitor/cell cycle block 
               
               
                 italics: protein inhibitor/activator 
               
               
                 *antihistamine 
               
               
                 #digoxin derivative 
               
               
                 +metabolic f/x 
               
               
                 !ion channel inhibitor/neuro R inhibitor 
               
             
          
         
       
     
       Example 5 
     Selected Compounds of Interest 
       [0142]    The present example demonstrates the utility of the present invention for providing a composition suitable for the inhibition of neuroblastoma, and for the treatment of neuroblastoma. 
         [0143]    Forty-seven (47) compounds were selected based on differential cell toxicity and compound mechanism of action. Forty are novel compounds for the treatment of neuroblastoma. None of these 40 compounds have been used clinically in neuroblastoma therapy nor have they been examined in clinical trials. Seven compounds have been previously used for neuroblastoma treatment (marked with asterisk), and serve as positive controls in the selection and screening process of new chemical entities that may be used in the treatment of neuroblastoma according the present invention. 
         [0000]    
       
         
               
               
               
             
           
               
                 TABLE 17 
               
               
                   
               
               
                   
                 NB12 
                   
               
               
                   
                 IC50 
               
               
                 Compounds of Interest: 
                 (nM) 
                 Notes: 
               
               
                   
               
             
             
               
                 2.3-Dimethoxy-1.4- 
                 nd 
                 ROS modulator/Redox cycling agent used 
               
               
                 naphthoquinone 
                   
                 to study role of ROS 
               
               
                 AKLAVINE 
                 778.5 
                 natural product; anti-infective; related 
               
               
                 HYDROCHLORIDE 
                   
                 structures have broad activity against NIH 
               
               
                   
                   
                 tumor lines 
               
               
                 Amodiaquin dihydrochloride 
                 790 
                 antimalarial; treatment of CNS degeneration 
               
               
                 dihydrate 
                   
                 (Alzheimer, MS); 4-aminoquinoline family; 
               
               
                   
                   
                 narrow therapeutic/toxic window in 
               
               
                   
                   
                 children; 4-Aminoquinolines depress 
               
               
                   
                   
                 cardiac muscle, impair cardiac conductivity, 
               
               
                   
                   
                 and produce vasodilatation with resultant 
               
               
                   
                   
                 hypotension 
               
               
                 AMSACRINE 
                 1214 
                 topo II inh; used in AML; may also be active 
               
               
                 HYDROCHLORIDE 
                   
                 vs malaria 
               
               
                 *ANCITABINE 
                 519.7 
                 cyclocytidine HCl; DNA-synthesis inhibitor 
               
               
                 HYDROCHLORIDE 
                   
                 (cytosine analog); antileukemic 
               
               
                 Azaguanine-8 
                 331 
                 purine analog 
               
               
                 beta-PELTATIN 
                 1949 
                 extracted from Mayapple rhizome (like 
               
               
                   
                   
                 podophyllotoxin); some evidence of in vitro 
               
               
                   
                   
                 anti-tumor f/x but vague 
               
               
                 Camptothecine (S.+) 
                 183.3 
                 topoisomerase 1 inh 
               
               
                 CGP-74514A hydrochloride 
                   
                 Cdk1 inh 
               
               
                 Chelerythrine chloride 
                 2553 
                 PKC inhibitor; affects translocation of PKC 
               
               
                   
                   
                 from cytosol to plasma membrane 
               
               
                 CHOLESTAN- 
                 2410 
                 Cholesterol oxidation product; cytotoxic due 
               
               
                 3beta.5alpha.6beta-TRIOL 
                   
                 to oxidative stress or cytoskeleton disruption 
               
               
                 CICLOPIROX OLAMINE 
                 2048 
                 topical antifungal, anti-inflammatory via 
               
               
                   
                   
                 inhibition of 5-lipoxygenase and cyclo- 
               
               
                   
                   
                 oxygenase; hydroxypyridone family; Loprox 
               
               
                 Clofazimine 
                 1417 
                 leprosy treatment; anti-inflammatory f/x; 
               
               
                   
                   
                 disrupts cc by binding DNA, may bind K+ 
               
               
                   
                   
                 transporters 
               
               
                 Colchicine 
                 29.3 
                 binds tubulin/blocks mitosis by preventing 
               
               
                   
                   
                 spindle formation; bioactive doses would 
               
               
                   
                   
                 probably be toxic 
               
               
                 CONVALLATOXIN 
                 73.17 
                 derived from lily of the valley; digitalis-like 
               
               
                   
                   
                 action 
               
               
                 CRASSIN ACETATE 
                 1947 
                 antineoplastic vs P388 leukemia and HT29 
               
               
                   
                   
                 colon cancer cells in vitro; cembranolides 
               
               
                   
                   
                 (14-member ring diterpenoid lactones) 
               
               
                   
                   
                 derived 
               
               
                   
                   
                 from Caribbean gorgonians (marine 
               
               
                   
                   
                 invertebrates) 
               
               
                 CRINAMINE 
                 1735 
                 HIF-1alpha inhibitor; affinity to the 
               
               
                   
                   
                 serotonin reuptake transport protein 
               
               
                 Dequalinium analog. C-14 linker 
                 1112 
                 Protein kinase C-alpha (PKC-alpha) 
               
               
                   
                   
                 inhibitor 
               
               
                 Dequalinium dichloride 
                 3617 
                 Selective blocker of apamin-sensitive K+ 
               
               
                   
                   
                 channels; mitochondria toxicity 
               
               
                 Digitoxin 
                 nd 
                 Na+/K+ pump inhibitor 
               
               
                 Digoxigenin 
                 nd 
                 Na+/K+ pump inhibitor 
               
               
                 Digoxin 
                 542.2 
                 Digitalis derivative; blocks Na+/K+ pump 
               
               
                 DIHYDROGAMBOGIC ACID 
                 1687 
               
               
                 Dihydroouabain 
                 1540 
                 Na+/K+ pump inhibitor 
               
               
                 ERYSOLIN 
                 3276 
                 organic isothiocyanate found in cruciferous 
               
               
                   
                   
                 veggies; increases accumulation of chemo 
               
               
                   
                   
                 drugs in PANC-1, MCF-7, NCI-H460 cell 
               
               
                   
                   
                 lines 
               
               
                 *Etoposide 
                 693.7 
                 topoisomerase II inh 
               
               
                 GAMBOGIC ACID 
                 1695 
                 principle pigment of gambage resin (bright 
               
               
                   
                   
                 orange); caspase activator (not well 
               
               
                   
                   
                 characterized); growth/tumor inhibitory vs 
               
               
                   
                   
                 HeLa, HEL, gastic cancer, lung carcinoma 
               
               
                   
                   
                 cell lines 
               
               
                 *Idarubicin 
                 203.7 
                 antineoplastic, DNA metabolism 
               
               
                 MECHLORETHAMINE 
                 438.2 
                 mustard gas derivative; polyfunctional 
               
               
                   
                   
                 alkylating agent = DNA breaks and 
               
               
                   
                   
                 crosslinks; non cell cycle phase specific 
               
               
                 MECLIZINE 
                 2537 
                 “antivert/bonine”; motion sickness/vertigo 
               
               
                 HYDROCHLORIDE 
                   
                 treatment; piperazine class of antihistamines 
               
               
                 MG 624 
                 848 
                 Nicotinic acetylcholine receptor antagonist; 
               
               
                   
                   
                 selectively inhibits alpha-bungarotoxin 
               
               
                   
                   
                 sensitive receptors that contain the alpha7 
               
               
                   
                   
                 subunit 
               
               
                 MITOXANTHRONE 
                 60.46 
                 topo II inh; used in ALL, breast cancer, non- 
               
               
                 HYDROCHLORIDE 
                   
                 hodgkin&#39;s lymphoma 
               
               
                 OUABAIN 
                 122.6 
                 Blocks movement of the H5 and H6 
               
               
                   
                   
                 transmembrane domains of Na+-K+ 
               
               
                   
                   
                 ATPases 
               
               
                 OXYBENDAZOLE 
                 nd 
                 benzimidazole anthelmintic used in horses 
               
               
                   
                   
                 and other ruminants 
               
               
                 Paclitaxel 
                 nd 
                 aka taxol; Antitumor agent; promotes 
               
               
                   
                   
                 assembly of microtubules and inhibits 
               
               
                   
                   
                 tubulin disassembly process 
               
               
                 Parthenolide 
                 2261 
                 feverfew extract; NFkB inh, p53 activ, 
               
               
                   
                   
                 increased ROS, JNK activ (indep of NFkB 
               
               
                   
                   
                 and ROS), inh of MAPK/ERK pathway; 
               
               
                   
                   
                 seems to work best as a 
               
               
                   
                   
                 chemosensitizer...studies in breast, skin, 
               
               
                   
                   
                 pancreatic, thoracic cell lines 
               
               
                 PATULIN 
                 nd 
                 polyketide lactone, produced by certain 
               
               
                   
                   
                 fungal species of Penicillium, Aspergillus 
               
               
                   
                   
                 and Byssochlamys growing on fruit, 
               
               
                   
                   
                 including apples, pears, grapes; crosslinks 
               
               
                   
                   
                 DNA, causes p38 and JNK phosphorylation 
               
               
                   
                   
                 in HEK cells 
               
               
                 PERIPLOCYMARIN 
                 2703 
                 digoxin relative 
               
               
                 PERUVOSIDE 
                 222.5 
                 inhibitor of Na+K+-ATPase; cardiac 
               
               
                   
                   
                 glycoside class 
               
               
                 *Podophyllotoxin 
                 135 
                 etoposide precursor/Antineoplastic 
               
               
                   
                   
                 glucoside; inhibitor of microtubule 
               
               
                   
                   
                 assembly; G2/M cc arrest 
               
               
                 Primaquine diphosphate 
                 nd 
                 antimalarial/inh of DNA, RNA, protein 
               
               
                   
                   
                 synthesis/muscarinic AChR inh 
               
               
                 Quinacrine dihydrochloride 
                 2556 
                 Monoamine oxidase (MAO) inhibitor; 
               
               
                   
                   
                 antimalarial 
               
               
                 Sanguinarine chloride 
                 1795 
                 Inhibitor of Mg2+ and Na+/K+-ATPase; 
               
               
                   
                   
                 isolated from the leaves and stems of 
               
               
                   
                   
                 Macleaya cordata and microcarpa 
               
               
                 *TENIPOSIDE 
                 705.5 
                 common NB treatment; semisynthetic 
               
               
                   
                   
                 podophyllotoxin derivative related to 
               
               
                   
                   
                 etoposide; topo II inh; induced single strand 
               
               
                   
                   
                 DNA breaks; activity in late S and G2 
               
               
                   
                   
                 phases 
               
               
                 TOMATINE 
                 nd 
                 alkaloid found in leaves of tomato and 
               
               
                   
                   
                 unripe fruit; tetrasaccharide tomato 
               
               
                   
                   
                 glycoalkaloid alpha-tomatine, trisaccharide 
               
               
                   
                   
                 beta(1)-tomatine, disaccharide gamma- 
               
               
                   
                   
                 tomatine, monosaccharide delta-tomatine, 
               
               
                   
                   
                 and their common aglycon tomatidine; 
               
               
                   
                   
                 inhibit the growth of human colon (HT29) 
               
               
                   
                   
                 and liver (HepG2) cancer cells 
               
               
                 *Vinblastine sulfate salt 
                 113 
                 Inhibitor of microtubule assembly 
               
               
                 *Vincristine sulfate 
                 61.95 
                 Inhibitor of microtubule assembly 
               
               
                   
               
             
          
         
       
     
       Example 6 
     Reduced Cytotoxicity to Non-Neuroblastoma Cells 
       [0144]    The standard of care for poor prognosis neuroblastoma tumors is intensive induction chemotherapy with cisplatin, etoposide, cyclophosphamide, and doxorubicin, high-dose myeloablative therapy with bone marrow transplant, surgery, radiation therapy, and biologic or maintenance therapy to eradicate minimal residual disease. The chemotherapy regimen is designed to induce massive genomic damage and subsequent cell death in proliferating cells. This strategy results in the death of both tumor and normal cells and is extremely debilitating to young patients. Additionally, this strategy does not target the tumor-initiating cells (TICs). The survival of NB TICs may contribute to tumor relapse. 
         [0145]    The compounds identified in the present invention target cellular pathways specific to neuroblastoma tumor-initiating cells while having little or no effect on normal cells ( FIG. 5 ). Therefore, the compositions and pharmaceutical preparations of the present invention will provide a treatment method for neuroblastoma having fewer and/or less intense or pronounced toxic side effects in patients. 
         [0146]    All documents, patents, journal articles and other materials cited in the present application are hereby incorporated by reference. 
         [0147]    Although the present invention has been fully described in conjunction with several embodiments thereof with reference to the accompanying drawings, it is to be understood that various changes and modifications may be apparent to those skilled in the art. Such changes and modifications are to be understood as included within the scope of the present invention as defined by the appended claims, unless they depart therefrom. 
       BIBLIOGRAPHY 
       [0148]    The references listed below as well as the references cited throughout the specification are incorporated herein by reference to the extent that they supplement, explain, provide a background for or teach methodology, techniques and/or compositions employed herein.
   1. Brodeur G M. (2003),  Nat Rev Cancer,  3:203-16.   2. Maris J M. (2005),  Curr Opin Pediatr,  17:7-13.   3. van Limpt V., et al. (2005),  Cancer Lett,  228:59-63.   4. Pardal, R., et al. (2003),  Nat Rev Cancer,  3:895-902.   5. Beachy, P. A., et al. (2004),  Nature,  432:324-31.   6. Warner, J. K., et al. (2004),  Oncogene,  23:7164.   7. Hamburger, A. &amp; Salmon, S. E. (1997),  J Clin Invest,  60:846-54.   8. Heppner, G. H. (1984),  Cancer Res , 44:2259-65.   9. Singh, S. K., et al. (2004),  Oncogene,  23:7267-73.   10. Al-Hajj, M. &amp; Clarke, M. F. (2004),  Oncogene,  23:7274-82.   11. Lapidot T, et al. (1994),  Nature,  367:645-8.   12. Bonnet D, Dick J E. (1997),  Nat Med,  3:730-7.   13. Al-Hajj, M., (2003),  Proc Natl Acad Sci USA,  100:3983-8.   14. Singh, S. K. et al. (2004),  Nature,  432:396-401.   15. van Noesel M M, et al. (1997),  Cancer,  80:834-43.   16. Toma, J. G. et al. (2001),  Nat Cell Biol,  3:778-84.   17. Fernandes K J, et al. (2004),  Nat Cell Biol,  6:1082-93.   18. Toma J G, et al. (2005),  Stem Cells,  23:727-37.   19. Christiansen J H, (2000),  Curr Opin Cell Biol,  12:719-24.   20. Ambros P F, et al. (2003),  Cancer Lett,  197:29-34.   21. Miettinen M, et al. (1998),  Am J Surg Pathol,  22:327-32.   22. LaBrosse E H, et al. (1976),  J Natl Cancer Inst,  57:633-8.   23. Barnabe-Heider F, Miller FD. (2003),  J Neurosci,  23:5149-60.   24. Marsh H N, et al. (2003),  J Cell Biol,  163:999-1010.   25. Lo Piccolo M S, Cheung N K, Cheung I Y. (2001),  Cancer,  92:924-31.   26. Fernandes K J, et al. (2006), Exp Neurol, 201:32-48.   27. Hafer R, et al. (1999),  J Neuroimmunol,  96:201-6.   28. Khanna C, et al. (2002),  In Vivo,  16:77-85.   29. Nakagawara A, Ohira M. (2004),  Cancer Lett,  204:213-24.   30. Ohira M, et al. (2005),  Cancer Cell,  7:337-50.   31. Weiss W A, et al. (1997),  EMBO J.  16:2985-95.   32. ElShamy W M, Fridvall L K, Ernfors P. (1998),  Neuron,  21:1003-15.   33. Lasorella A, et al. (2002)  Cancer Res , 62:301-6.   34. Valsesia-Wittmann S, et al. (2004),  Cancer Cell,  6:625-30.   35. Dubreuil V, et al. (2000),  Development,  127:5191-201.   36. Pattyn A, (2000),  Mol Cell Neurosci,  15:235-43.   37. Pozniak C D, (2000),  Science,  289:304-6.   38. Casciano I, et al. (2002),  Cell Death Differ,  9:246-51.   39. Matsumoto K, et al. (1995),  Cancer Res , 55:1798-806   40. Jaboin J, et al. (2002),  Cancer Res , 62:6756-63.   41. Kaplan D R, et al. (1993),  Neuron,  11:321-31.   42. Lucarelli E, et al. (1997),  Eur J Cancer,  33:2068-70.   43. Lavoie J F, et al. (2005),  J Biol Chem.,  280:29199-207.   44. Wartiovaara K, et al. (2002),  J Neurosci,  22:815-24.   45. Atwal J K, et al. (2000),  Neuron,  27:265-77.   46. Toma J G, et al. (2000),  J Neurosci,  20:7648-56.   47. Ellis J, Yao S. (2005),  Curr Gene Ther,  5:367-73.   48. Ellis J., (2005),  Human Gene Ther,  16:1241-6.   49. Mckenzie T A, et al. (2006),  J Neurosci,  26:6651-60.   50. Torkin R, et al. (2005),  Mol Cancer Ther,  4: 1-11.   51. Bamabe-Heider F, et al. (2005),  Neuron,  48: 253-65.   52. U.S. Pat. No. 6,787,355—Miller, et al. (2004).   53. Guzman, M L., et al. (2005),  Blood,  105(11): 4163-9.