Abstract:
A compound having the following structure:  
                         
 
     wherein Z is —CHOR 1 R 2  or —C(O)R 2 ; R 1  is H, an alkyl group, an aryl group, —OC(O)OR a , wherein R a  is an alkyl group, —C(O)R b  wherein R b  is an alkyl group, an aryl group, an alkoxy group, an amino group, an alkylamino group, a dialkylamino group, an aryl amino group, a diarylamino group, an arylalkyl amino group, a protecting group or a fluorous tag; R 2  is an alkyl group, an aryl group or an arylalkyl group; R 3  is H, an alkyl group, hydroxyalkyl group or an aryl group; R 4 —R 8  are independently the same or different and are hydrogen, an alkyl group, an alkenyl group, an alkynyl group, an aryl group, an alkoxy group, an aryloxy group, an acyloxy group, a haloalkyl group, a perfluoroalkyl group, fluorine, chlorine, bromine, a haloalkyloxy group, a carbamoyloxy group, a hydroxy group, a nitro group, a cyano group, a cyanoalkyl group, an azido group, an azidoalkyl group, a formyl group, a hydrazino group, a hydrazinoalkyl group, a hydroxyalkyl group, an alkoxyalkyl group, —NR 1 R m , wherein R 1  and R m  are independently hydrogen, an alkyl group, an aryl group, an arylalkyl group, or —C(O)R b , an aminoalkyl group, an alkylaminoalkyl group, a dialkylaminoalkyl group, an aryl aminoalkyl group, a diarylaminoalkyl group, an arylalkyl aminoalkyl group, —OC(O)OR a , wherein R a  is an alkyl group, —C(O)R b , —SR c , S(O)R c  or S(O 2 )R c  wherein R c  is hydrogen, —C(O)R b , an alkyl group, or an aryl group, (CH 2 ) n SiR d R e R f  wherein n is an integer within the range of 0 through 10 and R d , R e  and R f  are independently a C 1-10  alkyl group, a C 2-10  alkenyl group, a C 2-10  alkynyl group, an aryl group, a haloalkyl group, a cyanoalkyl group, an azidoalkyl group, a hydrazinoalkyl group, a hydroxyalkyl group, an alkoxyalkyl group, an aminoalkyl group, an alkylaminoalkyl group, a dialkylaminoalkyl group, an aryl aminoalkyl group, a diarylaminoalkyl group, an arylalkyl aminoalkyl group. Alternatively R 4  and R 5 , R 5  and R 6 ; R 6  and R 7 ; or R 7  and R 8  can form together a chain of 3 or four groups selected from CH, CH 2 , O, S, N, NH, N-alkyl or N-aryl. Provided that, at least one of R 5 —R 7  is not H, a lower alkyl group, fluorine, a cyano group, a hydroxyl group, hydroxyalkyl group, an alkoxy group, an aminoalkyl group, an alkylaminoalkyl group, a dialkylaminoalkyl group, an amino group, an alkylamino group, a dialkylamino group, a carbamoyloxy group, a formyl group or —C(O)R x  wherein R x  is an alkyl group.

Description:
CROSS-REFERENCE TO RELATED APPLICATION  
       [0001]    This application claims the benefit of U.S. Provisional Patent Application Serial No. 60/360,942, filed Mar. 1, 2002, the disclosure of which is incorporated herein by reference. 
     
    
     GOVERNMENTAL INTERESTS  
       [0002] This invention was made with government support under grant GM33372 and grant GM33678 awarded by the National Institutes of Health. The government has certain rights in this invention. 
     
    
     
       BACKGROUND OF THE INVENTION  
         [0003]    The present invention relates to novel mappicine analogs, to intermediates in the synthesis of mappicine compounds and to methods of synthesis of mappicine analogs and intermediates therefor.  
           [0004]    References set forth herein may facilitate understanding of the present invention or the background of the present invention. Inclusion of a reference herein, however, is not intended to and does not constitute an admission that the reference is available as prior art with respect to the present invention.  
           [0005]    Certain 1H-pyrano[3′,4′:6,7]indolizino[1,2-b]quinolinones, such as camptothecins, have been shown to have anticancer and antiviral activity. Indeed, a number of camptothecins are in use as anticancer agents. Although some camptothecins possess antiviral activity, they exhibit certain characteristics that are undesirable for antiviral agents and have thus not been used as antiviral agents. For example, camptothecins inhibit mammalian topoisomerase I, inhibit host cell DNA replication, and are cytotoxic to mammalian cells. camptothecins inhibit mammalian topoisomerase I, inhibit host cell DNA replication, and are cytotoxic to mammalian cells.  
           [0006]    It has been shown that substituted indolizino[1,2-b]quinolinones (and, in particular, mappicine ketones) that lack the α-hydroxylactone moiety of camptothecin are non-cytotoxic to mammalian cells, while exhibiting antiviral activity. In that regard, such substituted indolizino[1,2-b]quinolinones have been proposed for treating DNA viruses. See, for example, U.S. Pat. No. 5,883,255; Pendrak, I.; Barney, S.; Wittrock, R.; Lambert, D. M.; Kingsbury, W. D. “Synthesis and Anti-Hsv Activity of a-Ring-Deleted Mappicine Ketone Analog”  J. Org Chem.  1994, 59, 2623-2625; Pendrak, I.; Wittrock, R.; Kingsbury, W. D. “Synthesis and anti-HSV activity of methylenedioxy mappicine ketone analogs”  J. Org. Chem.  1995, 60, 2912-2915.  
           [0007]    A number of mappicine analogs are disclosed in U.S. Pat. No. 5,833,255; de Frutos, O.; Curran, D. P. “Solution phase synthesis of libraries of polycyclic natural product analogues by cascade radical annulation: Synthesis of a 64-member library of mappicine analogues and a 48-member library of mappicine ketone analogues”  J. Comb. Chem.  2000, 2, 639-649; Govindachari, T. R.; Ravindranath, K. R.; Viswanathan, N. “Isolation and Structure of Mappicine”  J. Chem. Soc., Perkin  1 1974, 1215-1221; Ome Das, Biswanath; Madhusudhan, Purushotham. “Chemoenzymatic synthesis of (S)- and (R)-mappicines and their analogs,”  Journal of Chemical Research, Synopses,  2000, 10, 476-477; Das, Biswanath; Madhusudhan, P., “Biochemical studies on natural products. V. Enantioselective synthesis of (S)- and (R)-mappicines and their analogues,”  Tetrahedron,  1999, 55(25), 7875-7880; Allaudeen, Hameed Sheik; Berges, David Alan; Hertzberg, Robert Philip; Johnson, Randall Keith; Kingsbury, William Dennis; Petteway, Stephen Robert, Jr. “Preparation of substituted indolizino[1,2-b]quinolinones” Published PCT Int. Appl. WO 9207856 1992; Dodds, Helen M.; Craik, David J.; Rivory, Laurent P., “Photodegradation of Irinotecan (CPT-11) in Aqueous Solutions: Identification of Fluorescent Products and Influence of Solution Composition,”  J. Pharm. Sci.,  1997, 86(12), 1410-1416; Sawada, Seigo; Muraji, Ko.,  Preparation of camptothecin derivatives as antitumor agents , Jpn. Kokai Tokkyo Koho (1992); Fortunak, J. M. D.; Mastrocola, A. R.; Mellinger, M.; Wood, J. L. “Preparation of mappicine ketones from camptothecins: Chemistry of the camptothecin E ring”  Tetrahedron Lett.  1994, 35, 5763-5764, the disclosures of which are incorporated herein by reference.  
           [0008]    Viruses are either DNA viruses or RNA viruses, but never both. DNA viruses can be divided into two groups: (1) those that have their genes on a double-stranded DNA molecule (dsDNA) (for example, smallpox); and (2) those that have their genes on a molecule of single-stranded DNA (ssDNA) (for example, Adeno-Associated Virus).  
           [0009]    RNA viruses can be divided into four groups: (1) those with a genome that consists of single-stranded antisense RNA; that is, RNA that is the complement of the message sense (also called negative-stranded RNA; examples include measles and Ebola); (2) those with a genome that consists of single-stranded sense RNA; that is, the RNA has message sense (can act as a messenger RNA—mRNA) (also called positive-stranded RNA; for example, poliovirus); (3) those with a genome made of several pieces of double-stranded RNA (for example, reovirus), and (4) retroviruses, in which RNA (also single-stranded) is copied by reverse transcriptase into a DNA genome within the host cell (for example, human immunodeficiency virus (HIV)).  
           [0010]    Retroviral Reverse Transcriptase. Retroviruses carry their genetic information as RNA, but must replicate through a double-strand DNA intermediate. Thus, following recognition and entry into a susceptible cell, the retroviral genomic RNA must be converted into viral DNA. Multiple steps are involved in this crucial step of replication, each of which is catalyzed by the retroviral enzyme reverse transcriptase (RT). This enzyme is therefore multifunctional, and possesses three enzymatic activities, RNA-dependent DNA polymerase activity (RDDP), DNA-dependent DNA polymerase activity (DDDP), and ribonuclease H activity (RNase H).  
           [0011]    Ribonuclease H(RNase H) is one of a family of enzymes termed nucleases, which act to hydrolyse nucleic acids. RNase H is unique among nucleases in that it selectively degrades the RNA component of an RNA/DNA duplex molecule, a double-strand nucleic acid comprised of one strand of ribonucleic acid (RNA) bound to a complementary strand of deoxyribonucleic acid (DNA) via Watson-Crick base pairing. Ribonucleases H are ubiquitous, found in virtually all organisms, as well in several types of viruses, including retroviruses and hepadnavirus.  
           [0012]    Ribonuclease H performs critical functions in the replication of several human pathogenic viruses, including retroviruses such as the human immunodeficiency virus (HIV) types 1 and 2, and the human T-cell leukemia viruses (HTLV) types 1 and 2. In addition, ribonuclease H is essential for the replication of the human hepadnavirus, hepatitis B virus (HBV).  
           [0013]    Several retroviruses are human pathogens. These include the human immunodeficiency viruses type 1 and 2 (HIV-1 and HIV-2), and the human T-cell leukemia viruses types 1 and 2 (HTLV-1 and HTLV-2). Of these, HIV-1 is by far the most serious pathogen. HIV-1 infection leads to AIDS, an incurable and inevitably fatal disease. Since identification of the virus in the early 1980&#39;s, it is estimated that more than 58 million individuals have been infected with HIV-1, and of these nearly 25 million have died of AIDS. HIV-1 infection remains one of the most serious infectious disease problems worldwide.  
           [0014]    A variety of biological agents are currently in use for the treatment of HIV-1 infections. HIV-1 RT has been, and remains, an important target for antiviral development. Many inhibitors of HIV-1 RT have been discovered, including nucleoside reverse transcriptase inhibitors (NRTI) such as 3′-azido-3′-deoxythymidine (AZT) and 2′,3′-dideoxy-3′-thiacytidine (3TC) and normucleoside reverse transcriptase inhibitors (NNRTI) such as nevirapine, delavirdine and efavirenz. However, virtually all inhibitors of HIV-1 RT are directed against the RDDP and/or DDDP activity of RT. Very few inhibitors of the ribonuclease H activity of HIV-1 (and HIV-2) reverse transcriptase have been described, and none are in clinical use.  
           [0015]    Although current therapeutics are initially very effective at controlling the course of HIV spread in an infected individual, thereby improving the quality of life and longevity of HIV-infected patients, prolonged therapy inevitably leads to viral resistance to these drugs. Resistance to RT inhibitors correlates with mutations in RT, and resistance to protease inhibitors correlates with mutations in the HIV protease. Clinical appearance of drug-resistant HIV imparts an unfavorable prognosis. In addition, the transmission of drug-resistant HIV variants from an infected treated individual to a previously naive individual is a serious problem. Drug therapies for use by these newly infected patients are restricted because of the infection by drug resistant virus.  
           [0016]    There is therefore an urgent need to identify new inhibitors of HIV replication, especially inhibitors that act on new viral targets, not presently targeted by current chemotherapies. These new targets include the ribonuclease H activity associated with the viral reverse transcriptase. Identification of compounds that inhibit HIV-1 reverse transcriptase associated RNase H has been identified as a research priority by a number of organizations, including the United States National Institutes of Health (NIH).  
           [0017]    Hepadnaviral Reverse Transcriptase. Human hepatitis B virus (HBV) is a major worldwide health threat and is responsible for the majority of the 1 to 2 million deaths annually from hepatitis. HBV is a member of the hepadnavirus family.  
           [0018]    Hepadnaviruses are small enveloped DNA viruses that replicate through an RNA intermediate. This replication mechanism therefore requires reverse transcription, to convert the RNA intermediate into viral DNA, a process carried out by the hepadnaviral P protein or reverse transcriptase. As is the case with retroviral reverse transcriptases, hepadnaviral P protein must be multifunctional to carry out reverse transcription. Thus, the protein possesses RNA-directed DNA polymerase and DNA-directed DNA polymerase activities, and ribonuclease H activity.  
           [0019]    There are very few treatments available for HBV infection. These include interferon therapy or liver transplantation, both of which are expensive and at best only partially successful. Recently, the nucleoside analog 3TC has been approved for treatment of chronic infection and transplant patients. This nucleoside is directed against the DNA polymerase activity of the HBV DNA polymerase (hepadnaviral P protein). Additional therapies need to be developed. The hepadnaviral P protein-associated ribonuclease H provides a target for this development.  
           [0020]    Objects of the present invention thus include development of reverse transcriptase inhibitors, development of RNase H inhibitors and development of improved methods of treatment of retroviruses, including HIV, and hepadnaviruses, including hepatitis B virus.  
           [0021]    Moreover, objects of the present invention also include development of methods of synthesizing large libraries of compounds for screening for such activities as well as other biological activities.  
         SUMMARY OF THE INVENTION  
         [0022]    In general, it has been discovered that analogs of the natural product mappicine (or mappicine analogs) inhibit retroviral reverse transcriptase and/or hepadnaviral reverse transcriptase by, for example, inhibiting the RNA-dependent DNA polymerase activity of reverse transcriptase and/or inhibiting the RNase H activity of reverse transcriptase (for example, HIV reverse transcriptase). Indeed, certain mappicine analogs were found to exhibit inhibition of the enzyme RNase H with a potency comparable to or better than the best currently known inhibitors. In that regard, the mappicine analogs of the present invention are suitable for use in a method of inhibiting retroviral reverse transcriptase in a patient (for example, a person or a mammal) infected with a retrovirus or hepadnavirus including the step of treating the patient with a pharmaceutically effective amount of the biologically active mappicine analog or a pharmaceutically acceptable salt thereof. Likewise, the mappicine analogs of the present invention are also suitable for use in a method of treating a patient infected with a retrovirus or hepadnavirus with a pharmaceutically effective amount of the mappicine analog or a pharmaceutically acceptable salt thereof.  
           [0023]    Highly active compounds of the present invention include, but are not limited to, 7-(1-Hydroxyethyl)-8-methyl-12-phenyl-2-trifluoromethoxy-11H-indolizino[1,2-b]quinolin-9-one, 12-Butyl-7-(1-hydroxyethyl)-8-methyl-2-trifluoromethoxy-11H-indolizino[1,2-b]quinolin-9-one, 7-(1-Hydroxyethyl)-2,8-dimethyl-1 2-phenyl-11H-indolizino[1,2-b]quinolin-9-one, 7-(1-Hydroxybutyl)-8-methyl-2-methylsulfanyl-12-pentyl-11H-indolizino[1,2-b]quinolin-9-one, 7-(1-Hydroxybutyl)-8-methyl- 12-phenyl-11H-indolizino [1,2-b]quinolin-9-one, 7-(1-Hydroxybutyl)-2,8-dimethyl-1 2-phenyl-11H-indolizino[1,2-b]quinolin-9-one, 4-Fluoro-7-(1-hydroxypropyl)-8-methyl-12-pentyl-1 1H-indolizino [1,2-b]quinolin-9-one, 7-(1-Hydroxypropyl)-2,8-dimethyl-1 2-pentyl-11H-indolizino[1,2-b]quinolin-9-one, 7-(1-Hydroxypropyl)-8-methyl-2-methylsulfanyl-1 2-pentyl-11H-indolizino[1,2-b]quinolin-9-one, 2-Ethyl-7-(1-hydroxy-2-methylbutyl)-8-methyl-1 2-pentyl-11H-indolizino[1,2-b]quinolin-9-one, 7-(1-Hydroxy-2-methylbutyl)-8-methyl-1 2-phenyl-11H-indolizino[1,2-b]quinolin-9-one, 7-(1-Hydroxy-2-methylbutyl)-8-methyl-2-methylsulfanyl-1 2-phenyl-11H-indolizino [1,2-b]quinolin-9-one, 1 2-Ethyl-7-(1-hydroxy-2-methylpropyl)-8-methyl-2-trifluoromethyl-11H-indolizino[1,2-b]quinolin-9-one, 12-Butyl-7-(1-hydroxy-2-methylpropyl)-2-methoxy-8-methyl-1H-indolizino[1,2-b]quinolin-9-one, 7-(Cyclohexylhydroxymethyl)-2-fluoro-8-methyl-12-pentyl-11H-indolizino[1,2-b]quinolin-9-one, 7-(Cyclohexylhydroxymethyl)-8-methyl-12-phenyl-2-trifluoromethyl-11H-indolizino[1,2-b]quinolin -9-one, 7-(Cyclohexylhydroxymethyl)-8-methyl-1 2-phenyl-2-trifluoromethoxy-11H-indolizino[1,2-b]quinolin-9-one, 2-Amino-12-ethyl-7-(1-hydroxypropyl)-8-methyl-11H-indolizino[1,2-b]quinolin-9-one, [7-(1-Hydroxypropyl)-8-methyl-9-oxo-12-(trimethylsilanyl)-9, 11-dihydroindolizino[1,2-b]quinolin-2-yl]-carbamic acid tert-butyl ester.  
           [0024]    Examples of retroviral infections of humans that can be treated with the mappicine compounds of the present invention include the human immunodeficiency viruses HIV-1 and HIV-2 and human T-cell leukemia virus (HTLV-1 and HTLV-2). Treatable retroviral infections of nonhumans include, for example, feline immunodeficiency virus, feline leukemia virus (cats), bovine immunodeficiency virus, bovine leukemia virus (cattle), equine infectious anemia virus (horses), caprine arthritis-encephalitis virus (goats), and Rous sarcoma virus infection of chickens.  
           [0025]    Examples of hepadnaviral infections of humans that can be treated with the mappicine compounds of the present invention include human hepatitis B virus (HBV).  
           [0026]    The mappicine analogs of the present invention can also be used in other treatments as, for example, described in U.S. Pat. No. 5,883,255.  
           [0027]    As used herein, the term “mappicine analog” refers generally to a compound possessing the 11H-indolizino[1,2-b]quinolin-9-one ring skeleton. The analog can have substantially any organic substituent or functional group substituted in place of one or more of the hydrogen atoms on the ring skeleton. The analog can also have a maximum of one additional fused ring generated by replacing two hydrogens by a chain of atoms or groups selected from CH, CH 2 , O, S, N, NH, N-alkyl or N-aryl. Preferred sizes of this additional ring are 5, 6, and 7.  
                         
 
           [0028]    For example, mappicine analogs of the present invention can have the-following general formulas:  
                         
 
           [0029]    These compounds can also be represented with the following general formula:  
                         
 
           [0030]    In general, the present invention provided compounds of formula (3) wherein Z is —CHOR 1 R 2  or —C(O)R 2    
           [0031]    wherein, R 1  is H, an alkyl group, an aryl group, —OC(O)OR a , wherein R a  is an alkyl group, —C(O)R b  wherein R b  is an alkyl group, an aryl group, an alkoxy group, an amino group, an alkylamino group, a dialkylamino group, an aryl amino group, a diarylamino group, or an arylalkyl amino group;  
           [0032]    R 2  is alkyl, aryl or arylalkyl;  
           [0033]    R 3  is H, alkyl, hydroxyalkyl or aryl;  
           [0034]    R 4 , R 5 , R 6 , R 7 , and R 8  are independently, the same or different, and are hydrogen, an alkyl group, an alkenyl group, an alkynyl group, an alkoxy group, an aryloxy group, an acyloxy group, a haloalkyl group, a perfluoroalkyl group, fluorine, chlorine, bromine, a carbamoyloxy group, a hydroxy group, a nitro group, a cyano group, a cyanoalkyl group, an azido group, an azidoalkyl group, a formyl group, a hydrazino group, a hydrazinoalkyl group, a hydroxyalkyl group, an alkoxyalkyl group, —NR 1 R m , wherein R 1  and R m  are independently hydrogen, an alkyl group, an aryl group, an arylalkyl group, or —C(O)R b , an aminoalkyl group, an alkylaminoalkyl group, a dialkylaminoalkyl group, an aryl aminoalkyl group, a diarylaminoalkyl group, an arylalkyl aminoalkyl group,  
           [0035]    —OC(O)OR a , wherein R a  is an alkyl group,  
           [0036]    —C(O)R b  wherein R b  is an alkyl group, an aryl group, an alkoxy group, an amino group, an alkylamino group, a dialkylamino group, an aryl amino group, a diarylamino group, an arylalkyl amino group,  
           [0037]    —SR c , S(O)R c  or S(O 2 )R c  wherein R c  is hydrogen, —C(O)R b , an alkyl group, or an aryl group, or  
           [0038]    (CH 2 ) n SiR d R e R f  wherein n is an integer within the range of 0 through 10 and R d , R e  and R f  are independently a C 1-10  alkyl group, a C 2-10  alkenyl group, a C 2-10  alkynyl group, an aryl group, a haloalkyl group, a cyanoalkyl group, an azidoalkyl group, a hydrazinoalkyl group, a hydroxyalkyl group, an alkoxyalkyl group, an aminoalkyl group, an alkylaminoalkyl group, a dialkylaminoalkyl group, an aryl aminoalkyl group, a diarylaminoalkyl group, an arylalkyl aminoalkyl group,  
           [0039]    or wherein R 4  and R 5 , R 5  and R 6 , R 6  and R 7 , or R 7  and R 8  form together a chain of three or four atoms or groups selected from CH, CH 2 , O, S, N, NH, N-alkyl or N-aryl.  
           [0040]    In one aspect, the present invention provides novel compounds of formula (3) as described above wherein R 8  is not H.  
           [0041]    In another aspect, the present invention provided compounds of formula (3) as described above, wherein at least one of R 5 —R 7  is not H, a lower alkyl group, fluorine, a cyano group, a hydroxyl group, a nitro group, hydroxyalkyl group, an alkoxy group, an aminoalkyl group, an alkylaminoalkyl group, a dialkylaminoalkyl group, an amino group, an alkylamino group, a dialkylamino group, a carbamoyloxy group, a formyl group or —C(O)R x  wherein R x  is an alkyl group.  
           [0042]    In a further aspect, the present invention provides compounds of formula (3) as described above provided that, when R 2  is an alkyl group and R 3  is a methyl group (or, in another embodiment, when R 3  is an alkyl group), R 4  is not H, an alkyl group, an aryl group, an aryloxy group, a nitro group, a cyano group, a hydroxyalkyl group, an alkoxyalkyl group, an aminoalkyl group, an alkylaminoalkyl group, an arylalkyl aminoalkyl group, —(CH 2 ) n SiR d R e R f  wherein n is an integer within the range of 0 through 10 and R d , R e  and R f  are independently an alkyl group.  
           [0043]    In a further aspect, the present invention provides compounds of formula (3) as described above provided that, when R 2  is an alkyl group and R 3  is a methyl group (or, in another embodiment, when R 3  is an alkyl group), R 5  is not H, an alkoxy group, an acyloxy group, fluorine, chlorine, bromine, a hydroxy group, an alkoxyalkyl group, —NR 1 R m  wherein R 1  and R m  are independently hydrogen or an alkyl group, an aminoalkyl group, an alkylaminoalkyl group or a dialkylaminoalkyl group.  
           [0044]    In a further aspect, the present invention provides compounds of formula (3) as described above provided that, when R 2  is an alkyl group and R 3  is a methyl group (or, in another embodiment, when R 3  is an alkyl group), R 6  is not hydrogen, an alkyl group, an alkoxy group, fluorine, a carbamoyloxy group, a hydroxyl group, a cyano group, a formyl group, a hydroxyalkyl group, an alkoxyalkyl group, —NR 1 R m  wherein R 1  and R m  are independently hydrogen or an alkyl group, an aminoalkyl group, an aminoalkyl group, an alkylaminoalkyl group, a dialkylaminoalkyl group, or —C(O)R b  wherein R b  is an alkyl group.  
           [0045]    In a further aspect, the present invention provides compounds of formula (3) as described above provided that, when R 2  is an alkyl group and R 3  is a methyl group (or, in another embodiment, when R 3  is an alkyl group), R 7  is not hydrogen, an alkoxy group a hydroxyl group or a cyano group.  
           [0046]    In the event that some combination of substituents creates a chiral center or another form of an isomeric center in any compound of the present invention, all forms of such isomer(s) are considered to be aspects of the present inventions. When a compound of the present invention contains a chiral center, the present invention includes the racemic mixture, the pure enantiomers, and any enantiomerically enriched mixture thereof.  
           [0047]    The present invention also provides novel intermediates in the synthesis of the above compounds. For example, the present invention provides compounds having the following structure:  
                         
 
           [0048]    wherein, X is hydrogen, a trialkylsilyl group (—SIR 10 OR 11 R 12 , wherein R 10 , R 11 , and R 12  are independently the same or different an alkyl group) or a radical precursor;  
           [0049]    R 1  is H, an alkyl group, an aryl group, —OC(O)OR a , wherein R a  is an alkyl group, —C(O)R b  wherein R b  is an alkyl group, an aryl group, an alkoxy group, an amino group, an alkylamino group, a dialkylamino group, an aryl amino group, a diarylamino group, an arylalkyl amino group, a protecting group or a fluorous tag;  
           [0050]    R 2  is an alkyl group, an aryl group or an arylalkyl group;  
           [0051]    R 3  is H, an alkyl group, hydroxyalkyl group or an aryl group; and  
           [0052]    R 4  is hydrogen, an alkyl group, an alkenyl group, an alkynyl group, an alkoxy group, an aryloxy group, an acyloxy group, a haloalkyl group, a perfluoroalkyl group, fluorine, chlorine, bromine, a carbamoyloxy group, a hydroxy group, a nitro group, a cyano group, a cyanoalkyl group, an azido group, an azidoalkyl group, a formyl group, a hydrazino group, a hydrazinoalkyl group, a hydroxyalkyl group, an alkoxyalkyl group, an amino group, an alkylamino group, a dialkylamino group, an aryl amino group, a diarylamino group, an arylalkyl amino group, an aminoalkyl group, an alkylaminoalkyl group, a dialkylaminoalkyl group, an aryl aminoalkyl group, a diarylaminoalkyl group, an arylalkyl aminoalkyl group,  
           [0053]    —OC(O)OR a , wherein R a  is an alkyl group,  
           [0054]    —C(O)R b  wherein R b  is an alkyl group, an aryl group, an alkoxy group, an amino group, an alkylamino group, a dialkylamino group, an aryl amino group, a diarylamino group, an arylalkyl amino group,  
           [0055]    —SR c , S(O)R c  or S(O 2 )R c  wherein R c  is hydrogen, —C(O)R b , an alkyl group, or an aryl group, or  
           [0056]    (CH 2 ) n SiR d R e R f  wherein n is an integer within the range of 0 through 10 and Rd, Re and R f  are independently a C 1-10  alkyl group, a C 2-10  alkenyl group, a C 2-10  alkynyl group, an aryl group, a haloalkyl group, a cyanoalkyl group, an azidoalkyl group, a hydrazinoalkyl group, a hydroxyalkyl group, an alkoxyalkyl group, an aminoalkyl group, an alkylaminoalkyl group, a dialkylaminoalkyl group, an aryl aminoalkyl group, a diarylaminoalkyl group, an arylalkyl aminoalkyl group.  
           [0057]    The present invention also provides compounds as set forth above wherein in the case that R 1  is H, R 2  is not an alkyl group or an arylalkyl group. The present invention also provides compounds as set forth above wherein in the case that R 2  is an alkyl group or an arylalkyl group, R 1  is not H.  
           [0058]    The present invention also provides compounds having the following structure:  
                         
 
           [0059]    wherein X is hydrogen, a trialkylsilyl group, or a radical precursor;  
           [0060]    R 1  is H, an alkyl group, an aryl group, —OC(O)OR a , wherein R a  is an alkyl group, —C(O)R b  wherein R b  is an alkyl group, an aryl group, an alkoxy group, an amino group, an alkylamino group, a dialkylamino group, an aryl amino group, a diarylamino group, an arylalkyl amino group, a protecting group or a fluorous tag;  
           [0061]    R 2  is an alkyl group, an aryl group or an arylalkyl group; and  
           [0062]    R 3  is H, an alkyl group, hydroxyalkyl group or an aryl group. The present invention further provided compounds as set forth above wherein in the case that R 1  is H, R 2  is not a C 1  or C 2  alkyl group and R 3  is not —CH 3 . Likewise, the present invention provides such compounds wherein in the case that R 1  is H, R 2  is not an alkyl group and R 1  is not an alkyl group.  
           [0063]    The present invention also provides compounds as set forth above wherein in the case that R 1  is H, R 2  is not an alkyl group or an arylalkyl group. The present invention also provides compounds as set forth above wherein in the case that R 2  is an alkyl group or an arylalkyl group, R 1  is not H.  
           [0064]    In another aspect, the present invention provides a method of synthesizing the following compound:  
                         
 
           [0065]    wherein R 1  is H, an alkyl group, an aryl group, —OC(O)OR a , wherein R a  is an alkyl group, —C(O)R b  wherein R b  is an alkyl group, an aryl group, an alkoxy group, an amino group, an alkylamino group, a dialkylamino group, an aryl amino group, a diarylamino group, an arylalkyl amino group, a protecting group or a fluorous tag;  
           [0066]    R 2  is an alkyl group, an aryl group or an arylalkyl group;  
           [0067]    R 3  is H, an alkyl group, hydroxyalkyl group or an aryl group;  
           [0068]    R 4 -R 8  are independently the same or different and are hydrogen, an alkyl group, an alkenyl group, an alkynyl group, an alkoxy group, an aryloxy group, an acyloxy group, a haloalkyl group, a perfluoroalkyl group, fluorine, chlorine, bromine, a carbamoyloxy group, a hydroxy group, a nitro group, a cyano group, a cyanoalkyl group, an azido group, an azidoalkyl group, a formyl group, a hydrazino group, a hydrazinoalkyl group, a hydroxyalkyl group, an alkoxyalkyl group, an amino group, an alkylamino group, a dialkylamino group, an aryl amino group, a diarylamino group, an arylalkyl amino group, an aminoalkyl group, an alkylaminoalkyl group, a dialkylaminoalkyl group, an aryl aminoalkyl group, a diarylaminoalkyl group, an arylalkyl aminoalkyl group,  
           [0069]    —OC(O)OR a , wherein R a  is an alkyl group,  
           [0070]    —C(O)R 1  wherein R b  is an alkyl group, an aryl group, an alkoxy group, an amino group, an alkylamino group, a dialkylamino group, an aryl amino group, a diarylamino group, an arylalkyl amino group,  
           [0071]    —SR c , S(O)R c  or S(O 2 )R c  wherein R c  is hydrogen, —C(O)R b , an alkyl group, or an aryl group,  
           [0072]    (CH 2 ) n SiR d R e R f  wherein n is an integer within the range of 0 through 10 and R d , R e  and R f  are independently a C 1-10  alkyl group, a C 2-10  alkenyl group, a C 2-10  alkynyl group, an aryl group, a haloalkyl group, a cyanoalkyl group, an azidoalkyl group, a hydrazinoalkyl group, a hydroxyalkyl group, an alkoxyalkyl group, an aminoalkyl group, an alkylaminoalkyl group, a dialkylaminoalkyl group, an aryl aminoalkyl group, a diarylaminoalkyl group, an arylalkyl aminoalkyl group,  
           [0073]    or where R 5 ,R 6 ; R 6 ,R 7 ; or R 7 ,R 8  form together a chain of 3 or four groups selected from CH, CH 2 , O, S, N, NH, N-alkyl or N-aryl.  
           [0074]    The method includes the step of reacting a precursor compound having the formula:  
                         
 
           [0075]    wherein R 9  is CH 2 CH═CHR 4  or CH 2 C≡CR 4 , with an aryl isonitrile having the formula:  
                         
 
           [0076]    under conditions suitable to generate the corresponding radical from the precursor compound.  
           [0077]    The present invention also provided a method of synthesizing a compound of the formula:  
                         
 
           [0078]    wherein X is a radical precursor;  
           [0079]    R 1  is H, an alkyl group, an aryl group, —OC(O)OR a , wherein R a  is an alkyl group, —C(O)R b  wherein R b  is an alkyl group, an aryl group, an alkoxy group, an amino group, an alkylamino group, a dialkylamino group, an aryl amino group, a diarylamino group, an arylalkyl amino group, a protecting group or a fluorous tag;  
           [0080]    R 2  is an alkyl group, an aryl group or an arylalkyl group;  
           [0081]    R 3  is H, an alkyl group, hydroxyalkyl group or an aryl group; and  
           [0082]    R 9  is CH 2 CH≡CHR 4  or CH 2 C≡CR 4 .  
           [0083]    The method includes the step of reacting a compound of the formula;  
                         
 
           [0084]    with a compound having the formula YCH 2 CH═CHR 4  or YCH 2 C≡CR 4 , wherein Y is a leaving group;  
           [0085]    Still further, the present invention provides a method of synthesizing a library of different compounds having the general formula:  
                         
 
           [0086]    wherein R 1  is H, an alkyl group, an aryl group, —OC(O)OR a , wherein R a  is an alkyl group, —C(O)R b  wherein R b  is an alkyl group, an aryl group, an alkoxy group, an amino group, an alkylamino group, a dialkylamino group, an aryl amino group, a diarylamino group, an arylalkyl amino group or a protecting group;  
           [0087]    R 2  is an alkyl group, an aryl group or an arylalkyl group;  
           [0088]    R 3  is H, an alkyl group, hydroxyalkyl group or an aryl group;  
           [0089]    R 4 —R 8  are independently the same or different and are hydrogen, an alkyl group, an alkenyl group, an alkynyl group, an alkoxy group, an aryloxy group, an acyloxy group, a haloalkyl group, a perfluoroalkyl group, fluorine, chlorine, bromine, a carbamoyloxy group, a hydroxy group, a nitro group, a cyano group, a cyanoalkyl group, an azido group, an azidoalkyl group, a formyl group, a hydrazino group, a hydrazinoalkyl group, a hydroxyalkyl group, an alkoxyalkyl group, an amino group, an alkylamino group, a dialkylamino group, an aryl amino group, a diarylamino group, an arylalkyl amino group, an aminoalkyl group, an alkylaminoalkyl group, a dialkylaminoalkyl group, an aryl aminoalkyl group, a diarylaminoalkyl group, an arylalkyl aminoalkyl group,  
           [0090]    —OC(O)OR a , wherein R a  is an alkyl group,  
           [0091]    —C(O)R b  wherein R b  is an alkyl group, an aryl group, an alkoxy group, an amino group, an alkylamino group, a dialkylamino group, an aryl amino group, a diarylamino group, an arylalkyl amino group,  
           [0092]    —SR c , S(O)R c  or S(O 2 )R c  wherein R c  is hydrogen, —C(O)R b , an alkyl group, or an aryl group,  
           [0093]    (CH 2 ) n SiR d R e R f  wherein n is an integer within the range of 0 through 10 and R d , R e  and R f  are independently a C 1-10  alkyl group, a C 2-10  alkenyl group, a C 2-10  alkynyl group, an aryl group, a haloalkyl group, a cyanoalkyl group, an azidoalkyl group, a hydrazinoalkyl group, a hydroxyalkyl group, an alkoxyalkyl group, an aminoalkyl group, an alkylaminoalkyl group, a dialkylaminoalkyl group, an aryl aminoalkyl group, a diarylaminoalkyl group, an arylalkyl aminoalkyl group,  
           [0094]    or where R 5 ,R 6 ; R 6 ,R 7 ; or R 7 ,R 8  form together a chain of 3 or four groups selected from CH, CH 2 , O, S, N, NH, N-alkyl or N-aryl.  
           [0095]    The method includes the steps of reacting B mixtures of A precursor compounds having the formula:  
                         
 
           [0096]    wherein, in each of the B mixtures, there are A compounds, each having a different fluorous tag Rf. A and B are integers, and R 9  is CH 2 CH═CHR 1  or CH 2 C≡CR 4 . X is a radical precursor as described above.  
           [0097]    Optionally, the method can include the steps of dividing each of the resultant B mixture into C separate mixtures, wherein C is an integer;  
           [0098]    In the case that C separate mixtures are formed, the method further includes the steps of reacting each of the resulting B*C mixtures, with one of C different aryl isonitriles having the general formula:  
                         
 
           [0099]    under conditions suitable to generate the corresponding radical precursor compounds from the precursor compounds. Alternatively, one can react a single isonitrile with the B mixture of A compounds. The method further includes the steps of separating the compounds in each of the resulting mixtures (for example, B mixtures or B*C mixtures) based upon the differences in fluorous content of the different Rf groups and converting Rf in the separated compounds to R 1 .  
           [0100]    The terms “alkyl”, “aryl” and other groups refer generally to both unsubstituted and substituted groups unless specified to the contrary. Unless otherwise specified, alkyl groups are hydrocarbon groups and are preferably C 1 -C 15  (that is, having 1 to 15 carbon atoms) alkyl groups, and more preferably C 1 -C 10  alkyl groups, and can be branched or unbranched, acyclic or cyclic. The above definition of an alkyl group and other definitions apply also when the group is a substituent on another group (for example, an alkyl group as a substituent of an alkylamino group or a dialkylamino group). The term “aryl” refers to phenyl or naphthyl. As used herein, the terms “halogen” or “halo” refer to fluoro, chloro, bromo and iodo.  
           [0101]    The term “alkoxy” refers to —OR g , wherein R g  is an alkyl group. The term “aryloxy” refers to —OR h , wherein R h  is an aryl group. The term acyl refers to —C(O)R i . The term “alkenyl” refers to a straight or branched chain hydrocarbon group with at least one double bond, preferably with 2-15 carbon atoms, and more preferably with 2-10 carbon atoms (for example, —CH═CHR j  or —CH 2 CH═CHR j ). The term “alkynyl” refers to a straight or branched chain hydrocarbon group with at least one triple bond, preferably with 2-15 carbon atoms, and more preferably with 2-10 carbon atoms (for example, —C≡CR k  or —CH 2 —C≡CR k ). The terms “alkylene,” “alkenylene” and “alkynylene” refer to bivalent forms of alkyl, alkenyl and alkynyl groups, respectively.  
           [0102]    The groups set forth above, can be substituted with a wide variety of substituents to synthesize mappicine analogs retaining activity. For example, alkyl groups may preferably be substituted with a group or groups including, but not limited to, a benzyl group, a phenyl group, an alkoxy group, a hydroxy group, an amino group (including, for example, free amino groups, alkylamino, dialkylamino groups and arylamino groups), an alkenyl group, an alkynyl group, a halogen (for example, perfluoroalkyl) and an acyloxy group. In the case of amino groups (—NR 1 R m ), R 1  and R m  are preferably independently hydrogen, an acyl group, an alkyl group, or an aryl group. Acyl groups may preferably be substituted with (that is, R u  is) an alkyl group, a haloalkyl group (for example, a perfluoroalkyl group), an alkoxy group, an amino group and a hydroxy group. Alkynyl groups and alkenyl groups may preferably be substituted with (that is, R j  and R k  are preferably) a group or groups including, but not limited to, an alkyl group, an alkoxyalkyl group, an amino alkyl group and a benzyl group.  
           [0103]    The term “acyloxy” as used herein refers to the group —OC(O)R g .  
           [0104]    The term “alkoxycarbonyloxy” as used herein refers to the group —OC(O)OR g .  
           [0105]    The term “carbamoyloxy” as used herein refers to the group —OC(O)NR 1 R m .  
           [0106]    For purpose of, for example, effecting separation of mappicine analogs prepared as a library in a combinatorial or parallel synthesis, R 1  can also be a or fluorous tag as described above. As used herein, the terms “fluorous tagging” or “fluorous-tagged” refers generally to attaching a fluorous moiety or group (referred to as a “fluorous tagging moiety,” a “fluorous tagging group” or simply a “fluorous tag”) to a compound to create a “fluorous-tagged compound”. Preferably, the fluorous tagging moiety is attached via covalent bond. However, other effective attachments such as ionic bonding, chelation or complexation can also be used. Fluorous tagging moieties facilitate separation of fluorous tagged compounds from other compounds as a result of differences in the fluorous nature of the compounds. Especially useful are fluorous separation methods such as fluorous liquid-liquid extraction, fluorous solid-liquid extraction, and/or fluorous chromatography.  
           [0107]    As used herein, the term “fluorous”, when used in connection with an organic (carbon-containing). molecule, moiety or group, refers generally to an organic molecule, moiety or group having a domain or a portion thereof rich in carbon-fluorine bonds (for example, fluorocarbons, fluorohydrocarbons, fluorinated ethers and fluorinated amines). The terms “fluorous-tagged reagent” or “fluorous reagent,” thus refer generally to a reagent comprising a portion rich in carbon-fluorine bonds. As used herein, the term “perfluorocarbons” refers generally to organic compounds in which all hydrogen atoms bonded to carbon atoms have been replaced by fluorine atoms. The terms “fluorohydrocarbons” and “hydrofluorocarbons” include organic compounds in which at least one hydrogen atom bonded to a carbon atom has been replaced by a fluorine atom.  
           [0108]    Fluorous moieties and/or the attachment of fluorous moieties or tags to organic compounds are discussed for example, in U.S. Pat. Nos. 5,859,247, 5,777,121, and U.S. patent application Ser. Nos. 09/506,779, 09/565,087, 09/602,105, 09/952,188 and 09/877,944, the disclosures of which are incorporated herein by reference. Fluorous tags can include portions that are not rich in carbon-fluorine bonds such as, for example, a spacer group (for example, an alkylene (—(CH) n —) group) via which the tag is attached to a molecule.  
           [0109]    Fluorous tags suitable for use in the present invention include, for example, a perfluoroalkyl group, a hydrofluoroalkyl group, a fluorinated ether group or a fluorinated amine group.  
           [0110]    Perfluoroalkyl groups are preferably of 2 to 20 carbons. Hydrofluoroalkyl groups are preferably of 2 to 20 carbons and include up to one hydrogen atom for each two fluorine atoms. For example, perfluorinated ether groups can have the general formula —[(CF 2 ) n O(CF 2 ) y ] z CF 3 , wherein x, y and z are integers. Perfluorinated amine groups can, for example, have the general formula —[(CF 2 ) x′ (NR a′ )CF 2 ) y′ ] z′ CF 3 , wherein x′, y′ and z′ are integers and wherein R a′  can, for example, be CF 3  or (CF 2 ) n′ CF 3 , wherein n′ is an integer. Fluorinated ether groups and fluorinated amine groups suitable for use in the present invention need not be perfluorinated, however. Fluorinated ether groups are preferably of 3 to 20 carbons. Fluorinated amine groups are preferably of 4 to 20 carbons.  
           [0111]    Certain groups such as hydroxy groups, amino groups and/or other groups of certain compounds of the present invention and certain compounds used in the methods of the present invention can be protected using protective groups as known in the art. Such protective groups include, but are not limited to, —SiR 10 R 11 R 12  wherein R 10 , R 11 , and R 12  are independently the same or different an alkyl group (preferably a lower alkyl group) or an aryl group; CHR x OR y  where R x  is H or alkyl (preferably lower alkyl, and more preferably methyl) and R y  is alkyl (preferably lower alkyl) or CH 2 C 6 H 3 R n R o  wherein R n  and R o  are independently the same or different, ortho, meta or para H, alkyl (preferably lower alkyl), alkoxy, nitro, cyano, halo, phenyl, trifluoromethyl or azido; CH 2 CH 2 OR 13  where R 13  is alkyl, CH 2 CH 2 SiR 10 R 11 R 12  or CH 2 CCl 3 ; 2-tetrahydropyranyl; 4-methoxy-2-tetrahydropyranyl; 2-tetrahydrofuranyl; CH 2 SRP where RP is alkyl (preferably lower alkyl); CH 2 CH 2 Si R 10 R 11 R 12 ; a tert-butyl group; CH 2 C 6 H 3 R q R r  wherein R q  and R r  are independently the same or different, ortho, meta, or para H, alkyl (preferably lower alkyl), alkoxy, nitro, cyano, halo, phenyl, trifluoromethyl or azido; or —C(O)R 14  wherein R 14  is H, alkyl (preferably lower alkyl), haloalkyl, aryl, alkoxy or OCH 2 C 6 H 3 R s R t , wherein R s  and R t  are independently the same or different, ortho, meta, or para H, alkyl (preferably lower alkyl), alkoxy, nitro, cyano, halo, phenyl, trifluoromethyl or azido. Other suitable protecting groups as known to those skilled in the art are disclosed, for example, in Greene, T., Wuts, P. G. M., Protective Groups in Organic Synthesis, Wiley (1991) and other general references set forth below, the disclosures of which is incorporated herein by reference.  
           [0112]    The protecting groups may be present in any precursors and intermediates and should protect the functional groups concerned against unwanted secondary reactions, such as acylations, etherifications, esterifications, oxidations, solvolysis, and similar reactions. In certain cases, the protecting groups may, in addition to this protection, effect a selective course of reactions. It is a characteristic of protecting groups that they lend themselves readily, i.e. without undesired secondary reactions, to removal, typically by solvolysis, displacement, hydrolysis, reduction, photolysis or also by enzyme activity, for example under conditions analogous to physiological conditions, and that they are not present in the end-products. The specialist knows, or can easily establish, which protecting groups are suitable with the reactions mentioned hereinabove and hereinafter.  
           [0113]    The protection of functional groups by protecting groups, the protecting groups themselves, and their cleavage reactions are described for example in standard reference works, such as J. F. W. McOmie, “Protective Groups in Organic Chemistry”, Plenum Press, London and New York 1973, in T. W. Greene, “Protective Groups in Organic Synthesis”, Wiley, New York 1981, in “The Peptides”; Volume 3 (editors: E. Gross and J. Meienhofer), Academic Press, London and New York 1981, in “Methoden der organischen Chemie” (Methods of organic chemistry), Houben Weyl, 4th edition, Volume 15/1, Georg Thieme Vedag, Stuttgart 1974, in H.-D. Jakubke and H. Jescheit, “Aminosauren, Peptide, Proteine” (Amino adds, peptides, proteins), Verlag Chemie, Weinheim, Deerfield Beach, and Basel 1982, and in Jochen Lehmann, “Chemie der Kohlenhydrate: Monosaccharide und Derivate” (Chemistry of carbohydrates: monosaccharides and derivatives), Georg Thieme Verlag, Stuttgart 1974.  
           [0114]    In a preferred embodiment of the present invention, a fluorous tag is present in a protecting group. In general, any protective group can be rendered fluorous by, for example, replacing a hydrogen atom or a group with a fluorous tag. Examples of preferred fluorous protecting groups include, but are not limited, to —C(O)(CH 2 ) N Rf, wherein N is an integer from 2 to 5, —CH 2 C 6 H 4 (CH 2 ) N Rf, meta or para, wherein N is an integer from 0 to 5, —C 6 H 4 (CH 2 ) N Rf, meta or para, wherein N is an integer from 0 to 5 and  
                         
 
           [0115]    wherein N is an integer from 2 to 5 and R u  and R v  are independently, the same or different, an alkyl group or an aryl group.  
           [0116]    The term “radical precursor(s)” as used herein and as well known to those skilled in the art refers generally to those atoms or functional groups that cleave to generate radicals under standard conditions of chain or non-chain radical reactions. Common radical precursors include, but are not limited, halogens (typically except fluorine), carboxylic acids and derivatives thereof (such as thiohydroxamates), selenophenyl groups, diazonium salts, and the like. See, for example, Giese, B.  Radicals in Organic Synthesis: Formation of Carbon-Carbon Bonds ; Pergamon, Oxford (1986), the disclosure of which is incorporated herein by reference.  
           [0117]    A used in the present invention, the term “leaving group” refers to a part of a compound or molecule that is cleaved in a substitution reaction. Many different leaving groups suitable for use in the present invention are known to those skilled in the art. For the purposes of this invention, preferred leaving groups can, for example, molecules or ions whose conjugate acids have a pKa of less than about 18. Leaving groups whose conjugate acids have a pKa of less than about 10 are more preferred. Even more preferred are leaving groups whose conjugate acids have a pKa of less than about 5. Suitable leaving groups include, but are not limited to, a halide (for example, Cl, Br or I), alkane sulfonate (for example, mesylate) or arene sulfonate (for example, tosylate).  
           [0118]    For purpose of biological activity, R 1 , R 2 , R 3 , R 6 , R 7  and R 8  are, in general, preferably not excessively bulky to maintain the activity of the resultant mappicine analog. Preferably, therefore, R 1 , R 2 , R 3 , R 6 , R 7  and R 8  independently have a molecular weight less than approximately 350. More preferably R 1 , R 2 , R 3 , R 6 , R 7  and R 8  independently have a molecular weight less than approximately 250. In general, the total molecular weight of the sum of all R 1 , R 2 , R 3 , R 6 , R 7  and R 8  groups preferably does not exceed about 750, and more preferably does not exceed about 600. Certain intermediates, such as fluorous tagged mappicine compounds of the present invention need not satisfy the above criteria.  
           [0119]    Some of the mappicine analogs of the present invention can be prepared for pharmaceutical use as salts with inorganic acids such as, but not limited to, hydrochloride, hydrobromide, sulfate, phosphate, and nitrate. The mappicine analogs can also be prepared as salts with organic acids such as, but not limited to, acetate, tartrate, fumarate, succinate, citrate, methanesulfonate, p-toluenesulfonate, and stearate. Other acids can be used as intermediates in the preparation of the compounds of the present invention and their pharmaceutically acceptable salts. Likewise, for some analogs of the present invention, salts with organic (for example, amine) and inorganic (for example, sodium and potassium) bases can also be prepared.  
           [0120]    The compounds of the present invention can, for example, be administered by any conventional route of administration, including, but not limited to, intravenously, intramuscularly, orally, subcutaneously, intratumorally, intradermally, and parenterally. The pharmaceutically effective amount or dosage is preferably between 0.01 to 250 mg of one of the compounds of the present invention per kg of body weight. More preferably, the pharmaceutically effective amount or dosage is preferably between 0.1 to 40 mg of one of the compounds of the present invention per kg of body weight. In general, a pharmaceutically effective amount or dosage contains an amount of one of the compounds of the present invention effective to display antiretroviral behavior. Pharmaceutical compositions containing as an active ingredient one of the compounds of the present invention or a pharmaceutically acceptable salt thereof in association with a pharmaceutically acceptable carrier or diluent are also within the scope of the present invention.  
           [0121]    The present invention also provides a pharmaceutical composition comprising any of the compounds of the present invention and a pharmaceutically acceptable carrier. The composition may, for example, contain between 0.1 mg and 3 g, and preferably between approximately 0.1 mg and 500 mg of the compounds of the present invention, and may be constituted into any form suitable for the mode of administration. 
       
    
    
     BRIEF DESCRIPTION OF THE DRAWINGS  
       [0122]    [0122]FIG. 1A illustrates a synthetic scheme for synthesis of the 5 (AG) series of mappicine analogs of the present invention.  
         [0123]    [0123]FIG. 1B illustrates other synthetic scheme for synthesis of mappicine analogs.  
         [0124]    [0124]FIGS. 2A through 2M illustrates chemical structures of a number of mappicine analogs of the 5(3-8) series of the present invention.  
         [0125]    [0125]FIG. 3 illustrates a synthetic scheme for fluorous mixture synthesis of mappicine analogs of the present invention.  
         [0126]    FIGS.  4  illustrates a modified synthesis of an iodopyridine intermediate of the present invention.  
         [0127]    [0127]FIG. 5 illustrates Synthesis of propargyl bromides isonitriles intermediates.  
         [0128]    [0128]FIG. 6 illustrates the preparation of tagging alcohols.  
         [0129]    [0129]FIG. 7 illustrates mixture synthesis of tagged mappicines of the present invention.  
         [0130]    [0130]FIG. 8 illustrates HPLC analysis of tagged mappicines.  
         [0131]    [0131]FIG. 9 illustrates peparative demixing of tagged mappicines.  
         [0132]    [0132]FIG. 10 illustrates detagging of tagged mappicines and SPE purification.  
         [0133]    [0133]FIG. 11 illustrates a dose response curve for compound AG2M indicating inhibition of HIV-1 reverse transcriptase (RT).  
         [0134]    [0134]FIG. 12 illustrates a dose response curve for compound AG6M indicating inhibition of HIV-1 reverse transcriptase (RT).  
         [0135]    [0135]FIG. 13 illustrates in vitro dose-response curves for mappicine analogs 4.7.5 and 7.5.6 inhibition of HIV-1 reverse transcriptase associated RNase H activity.  
         [0136]    [0136]FIG. 14 illustrates antiviral and toxicity data for mappicine analog 7.5.6. 
     
    
     DETAILED DESCRIPTION OF THE INVENTION  
       [0137]    Recently, a fluorescence-based assay to perform screens in search of HIV RNase H inhibitors has been developed. The assay, which is carried out in 96-well microplates and is adaptable to robotics, is the first high-throughput screen for RNase H and is described in further detail in the Experimental section. HIV-1 RT and human RNase H were cloned, and thus comparative analysis of inhibitor action could be conducted simultaneously.  
         [0138]    In the course of the present studies, it was speculated that elongated “flat” structures may be desirable for binding to RNase H domain. For example, the compound N-4-(t-butylbenzoyl)-2-hydroxy-1-napthaldehyde hydrazone (or more conveniently BBNH) is one of the most active RNase H inhibiting compounds discovered to date with an IC 50 ≈2 μm (wherein IC 50  refers to the Inhibitory Concentration that provides 50% reduction in target activity). See, for example, Borkow, G. et al., “Inhibition of the Ribonuclease H and DNA Polymerase Activities of HIV-1 Reverse Transcriptase by N-(4-tert-Butylbenzoyl)-2-hydroxy-1-naphthaldehyde Hydrazone,”  Biochemistry  1997, 36, 3179-3185. Mappicine analogs similarly exhibit an elongated flat structure.  
         [0139]    There are many ways to make camptothecin and mappicine analogs and substantially any of these can be readily modified in accordance with the teachings herein to make the compounds of the present invention. Several representative examples of preferred synthetic routes to make the compounds of this invention are summarized below and in FIG. 1B. Compounds of the general formula I with X and R 3  as described above can be subjected to iodine/metal exchange and the resulting organometallic species (for example, a lithium or Grignard reagent) is contact with an aldehyde R 2 CHO to give II with R 1 ═H. Conversion of this compound to the other R 1  groups of this invention uses standard reactions. Also, I can be acylated, for example by Stille reaction with R 2 COSnBu 3 , to give IV, which can be used for onward reactions in a manner substantially similar to II or converted to II by standard reduction. For certain sequences, the conversion of II, X=TMS to II, X=I can be useful and can be accomplished by iododesilylation with, for example, ICl. Demethylation of II, for example with TMSI or HI, followed by alkylation with R 4 CCCH 2 Br, R 4 CH═CHCH 2 Br or related allylating or propargylating agents gives III, R 9 ═CH 2 CCR 4  or CH 2 CH═CHR 4 . See, for example, Liu, H.; Ko, S. B.; Josien, H.; Curran, D. P.  Tetrahedron Lett.  1995, 36, 8917-8920.  
         [0140]    Compounds III can be used in many ways to make the mappicines of the present invention. For example, reaction of III, R 9 ═CH 2 CCR 4  or CH 2 CH═CHR 4  and X=a radical precursor with isonitrile V with R 5 —R 8  as described above under conditions for cascade radical annulation (see FIG. 1B) provides mappicines VI. Preferred radical precursors are iodine and bromine. Reaction of III, X═Br or I, with V to give VI can also be promoted by certain transition metals susch as, for example, salts or complexes of palladium. See, for example, U.S. Patent Provisional Patent Application Serial No. 60/382,292, the disclosure of which is incorporated herein by reference. Reaction of III, R 9 ═H and X=chlorine, bromine, or iodine with VII, X═H and LG=a leaving group under conditions for N-alkylation provides VIII, which can in turn can be converted to VII under either radical conditions (for example, treatment with Bu 3 SnH) or organometallic conditions, for example, treatment with palladium catalysts. Likewise, reaction of III, R 9 ═H and X═H with VII, LG=leaving group and X=chlorine, bromine or iodine as above provides IX, which again can be converted to VII under radical or organometallic conditions. See, for example, Comins, D. L.; Hong, H.; Jianhua, G.  Tetrahedron Lett.  1994, 35, 5331-5334. Comins, D. L.; Hong, H.; Saha, J. K.; Gao, J. H.  J. Org. Chem.  1994, 59, 5120-5121. Comins, D. L.; Saha, J. K.  J. Org. Chem.  1996, 61, 9623-9624.  
         [0141]    Mappicines VII can be converted to mappicine ketones by using standard alcohol oxidations. In turn, if ketones such are IV are used in the synthetic sequence, mappicine ketones result directly, and these can be converted to mappicines by standard reductions.  
         [0142]    Libraries of mappicine analogs were studied in an HW RNase H assay. Mappicine analogs (see, for example, FIGS. 2A through 2M) of the present invention can, for example, be prepared via a parallel library synthesis via a cascade radical annulation method as disclosed in de Frutos, O.; Curran, D. P.  J. Comb. Chem.  2000, 2, 639, the disclosure of which is incorporated by reference. Mappicine analogs can also be prepared in a traditional (non-parallel) fashion as described below. In several of the representative non-parallel syntheses described below, 12- and/or 2-substituted (for example, 2-hydroxy, 2-AcO, 10-BocNH or 2-amino) mappicine analogs were prepared using the synthetic scheme set forth in FIG. 1A. Substitutions at corresponding positions in camptothecin analogs are, for example, known to enhance biological activity in certain camptothecin analogs.  
         [0143]    In the synthesis, intermediate 1 was prepared according to the reported procedure. de Frutos, O.; Curran, D. P.,  J. Comb. Chem.,  2000, 2, 639. Beginning with 1, the synthesis of, for example, 2-hydroxy and 2-amino mappicine analogs followed the sequence set forth below. As illustrated in FIG. 1, N-alkylation of iodopyridone 1 with the corresponding propargyl bromide 2 (R 4  is, for example, Et or TMS) provided the radical precursors 3a and 3b. Second, [4+1] cascade radical reaction of pyridones 3a/b with the corresponding isonitriles 4 (R 6  is, for example BocNH or AcO) gave rise to mappicine analogs 5-AG-2M, 5-AG-SM and 5-AG-7M. Subsequently, 2-AcO and 2-BocNH deprotection yielded 2-hydroxy and 2-amino mappicine analogs 5-AG-3M, 5-AG-6M and 5-AG-8M.  
         [0144]    The compounds of the present invention can also be synthesized via fluorous mixture synthesis, which is a homogeneous solution-phase mixture technique that provides the isolation of the individually pure components at the end of the synthesis.  
         [0145]    One embodiment of a general scheme for A*B*C compounds (for example, 7*8*10 or 560 compounds) is discussed below.  
         [0146]    In a representative fluorous mixture synthesis, seven pyridinyl alcohols bearing different R 2  groups were first attached to seven different tags and then mixed together (see FIG. 3). The mixture M-9 (the prefix “M” designates a mixture) underwent two consecutive one-pot reactions to generate a mixture of tagged pyridones M-10. The new mixture was then divided to eight portions for N-alkylation with eight propargyl bromides bearing different R 4  groups in parallel. The resulting eight mixtures M-12 were each split into ten portions for parallel free radical reactions with ten isonitriles bearing different R 5 —R 8  groups to generate eighty mixtures of tagged mappicines M-13. Each mixture containing seven components was then demixed by HPLC to give a total of 560 individual, pure mappicine analogs after detagging. This 4-step mixture synthetic process included two one-pot and two parallel reactions, and allowed combinatorialization of 3 sets of building blocks.  
         [0147]    An important starting material for the mixture synthesis is known iodopyridine 18, see de Frutos, O.; Curran, D. P. Solution phase synthesis of libraries of polycyclic natural product analogue by cascade radical annulation: Synthesis of a 64-member library of mappicine analogues and a 48-member library of mappicine ketone analogues.  J. Comb. Chem.  2000, 2, 639-649. The previous small-scale synthesis is not preferred because the deoxygenation of aldehyde 17 gave only 30% yield of 18 in gram-scale reactions (see FIG. 4). A new two-step sequence of the present invention involving primary alcohol 17 reliably gave 72% overall yield of iodopyridine 18 from aldehyde 16.  
         [0148]    The three sets of building blocks needed for the diversity plan were easily obtained. All seven aldehydes are commercially available. Eight propargyl bromides 19{1-8} were selected from commercial sources or prepared from the corresponding alcohols (see reaction A of FIG. 5). A total of ten aryl isonitriles 201-10} were prepared from the corresponding anilines (see reaction B of FIG. 5). Fluorous tags were prepared by addition of the lithium reagent derived from RfCH 2 CH 2 I to diisopropylchorosilane to give RfCH 2 CH 2 (iPr) 2 SiH.  
         [0149]    Seven alcohols 21{1-7} were prepared individually by quenching the Grignard reagent derived from iodopyridine 18 with seven different aliphatic aldehydes (see FIG. 6). Silyl triflates were generated from fluorous silanes and used in situ to protect alcohols 21{1-7} to give 9{1-7}. The tags in an order of increasing fluorine content were matched to the corresponding alcohols in an order of decreasing polarity of the R 2  side chain. In this way the best HPLC separation of the mixture will be achieved by matching the primary separation factor (fluorine content) of fluorous silica gel with a secondary separation factor (polarity). Seven alcohols with different R 2  substituents were coded with silyl tags as follows: Me/C 3 F 7 , Pr/C 4 F 9 , Et/C 6 F 13 , s-Bu/C 7 F 15 , i-Pr/C 8 F 17 , cyclohexyl/C 9 F 19 , cyclohexylethyl/C 10 F 21 . The tags for the propyl, ethyl, and s-butyl, isopropyl groups were deliberately mismatched to test if tag dominance overwhelms side chain polarity.  
         [0150]    The mixture synthesis of the 560-member mappicine library is summarized in FIG. 7. The numbering system for the intermediates and final products is that employed by the  Journal of Combinatorial Chemistry  published by the American Chemical Society (see “Instructions to Authors”); n{x,y,z}, where n is the number of the compound scaffold, x is the number of the R 2  substituent and its associated tag Rf {1,7} derived from pyridone 9, y is the number of the R 4  substituent {1-8} derived from propargyl bromide 19, and z is the number of the R 6  or R 8  substituent {1-10} derived from the isonitrile 20. There are accordingly 7*8*10=560 tagged mappicines 13 and 560 corresponding final mappicines 5. By way of example, tagged mappicine 13{4,3,5} has R 2 =s-Bu and Rf=C 7 F 15 {4}, R 4 =Me {3}, and R 6 =Et.  
         [0151]    Equimolar amounts of the seven-tagged compounds 9{1-7} were mixed and subjected to iodonative desilylation with ICI. The crude product mixture 10{1-7} was demethylated with boron tribromide, and the resulting mixture of seven pyridones 11{1-7} was purified by standard silica gel column chromatography. The purified mixtures were then split into eight portions and each portion (2.1 mmol) was subjected to N-alkylation with 1.25 equiv of propargyl bromides 19{1-8}. After flash chromatographic purification on silica gel, each of the eight mixtures of N-propargyl pyridones 12{1-7,1-8} was split into ten portions (0.15 mmol each) and irradiated under a sunlamp with 3.0 equiv of aryl isonitriles 20{1-10} and catalytic amount of hexamethylditin. These eighty crude mixtures were purified by rapid solid-phase extraction (SPE) with normal silica gel. Unreacted N-propargyl pyridones 12{1-7,1-8} and isonitriles 20{1-10} and other byproducts were washed off with 10% EtOAc/hexanes and fluorous-tagged mappicine mixtures 13{1-7,1-8,1-10} were eluted with 15% MeOH/EtOAc. All eighty mixtures were analyzed by automated LC-MS before loading onto a semi-preparative HPLC column for demixing.  
         [0152]    A typical LC trace for the analytical demixing of 13{1-7,4,3} is shown in FIG. 8 (UV detection, upper trace; MS detection, lower trace). Besides the solvent front, seven well-resolved peaks were detected in the UV channel in this and each of the other 79 mixtures. As revealed by the mass spectroscopy, the seven compounds are the expected tagged mappicines, which eluted in the order of increasing fluorine content of the tag despite the deliberate reversal of four of the side chains. The molecular ions of all of the expected 560 fluorous-tagged mappicines were detected by LC-MS. This result is a powerful demonstration that the elution order of the products can be predicted from the initial fluorous tagging of the starting substrates. To illustrate the analytical separation of the mixture of seven compounds, the retention times of 56 out of the 560 tagged mappicines are listed in Table 1. All of the seven peaks of mixtures 13{1-7,1-8,4} and 13{1-7,1-8,7} eluted about 2 min slower than the rest of the library components. This is believed to be a result of the presence of the extra CF 3  group in the A-ring of mappicine in these mixtures.  
                                                                                   TABLE 1                           Retention Times (min) of 56 of the 560 Tagged Mappicines 13. a              13{1-7, y, z}   C 3 F 7     C 4 F 9     C 6 F 13     C 7 F 15     C 8 F 17     C 9 F 19     C 10 F 21                      y, z = {1, 1}   3.5   4.5   6.7   8.7   10.7   12.3   15.7       {2, 2}   4.0   5.3   7.8   10.0   11.8   13.3   16.4       {3, 3}   3.6   4.6   6.7   8.6   10.7   12.2   15.5       {4, 4} b     5.3   7.0   9.8   12.0   13.9   15.2   18.0       {5, 5}   4.6   6.0   8.8   11.0   13.3   14.9   19.7       {6, 6}   4.4   5.7   8.3   10.4   12.4   13.8   16.8       {7, 7} b     6.9   8.7   11.7   13.8   15.6   16.8   19.8       {8, 8}   3.7   4.8   7.1   9.1   11.2   12.7   15.9                                  
 
         [0153]    A semi-preparative Fluophase-RP column (20×250 mm, 5 μm) was used for demixing of eighty tagged mappicine mixtures. Samples containing about 0.07 mmol (45-65 mg) of 7-component mixtures were dissolved in 300-350 μL of THF, injected onto the column and demixed into their seven individual components. A typical chromatogram of demixing along with conditions for the separation of 13{1-7,6,2} are shown in FIG. 9.  
         [0154]    The fluorous silyl protecting groups were cleaved with HF-pyridine in THF (see FIG. 10). The short tags (C 3 F 7  and C 4 F 9 ) can be easily detached within 1 h at 60° C., while longer tags required extended heating times up to 10 h. The crude products were partitioned between AcOEt and H 2 O. The concentrated organic layers were loaded onto reverse phase SPE cartridges and eluted with MeOH/H 2 O (80/20) or THF/H 2 O (50/50). Mappicines 5 eluted first. Cleaved tags (silanols) followed when the cartridges were washed with MeOH or THF.  
         [0155]    Finally, the products were quantified by weighing, and the amount of 560 mappicines was distributed as follows: 315 samples (56%) were between 1-2 mg, 180 samples (32%) were less than 1 mg, and 65 samples (12%) were greater than 2 mg.  
         [0156]    Since the structures of all 560 compounds were characterized by LC-MS before demixing, the final analyses were focused on the assessment of product purity. Several  19 F NMR analyses were carried out to ensure that there were no tag residues after detagging and SPE. HPLC analysis with UV detection of 112 (20% of the library) randomly selected samples indicated that the product purity were greater than 90%. Additional product characterizations included MS/LC-MS and  1 H NMR/LC-NMR analyses (see Examples section).  
         [0157]    All the mappicine samples had the expected substituents with one exception. HPLC analysis of those 56 mappicine analogs 13{1-7,1-8,10} with the MeS-substituent at the A ring gave more than one peak. MS analysis revealed that one of the peaks was the expected sulfide and the others (usually one peak, sometimes two) were the sulfoxides as a mixture of diastereomers.  
         [0158]    Assay results for a number of mappicine analogs synthesized as described above are set forth in Tables 2 and 3 below. The designations set forth in FIG. 2 for the corresponding chemical structures are used in Table 2. The assay results provided in Table 2 were obtained at 10 μM inhibitor concentrations. The structures of the analogs in Table 2 are illustrated in FIGS. 1A and 2A through  2 M. The designations set forth in Table 3 use the convention employed by the  Journal of Combinatorial Chemistry  as set forth above (n{x,y,z}). The assay results provided in Table 3 were obtained at 20 μM inhibitor concentrations.  
                                                               TABLE 2                                       Residual activity (%)                    Compounds   Polymerase   RNase H                            5 (3Aε)   34   99           5 (3Aδ)   32   104           5 (3Aβ)   54   95           5 (3Aα)   90   97           5 (3Bε)   28   91           5 (3Bδ)   81   99           5 (3Bβ)   78   75           5 (3Bα)   54   83           5 (3Cε)   76   101           5 (30δ)   74   93           5 (30β)   70   84           5 (3Cα)   64   96           5 (3Dε)   61   79           5 (3Dδ)   60   67           5 (3Dβ)   85   70           5 (3Dα)   69   78           5 (4Aε)   83   93           5 (4Aδ)   45   95           5 (4Aβ)   71   101           5 (4Aα)   67   104           5 (4Bε)   61   89           5 (4Bδ)   44   90           5 (4Bβ)   49   113           5 (4Bα)   98   108           5 (4Cε)           5 (4Cδ)   61   109           5 (4Cβ)   67   94           5 (4Cα)   94   91           5 (4Dε)   49   76           5 (4Dδ)   51   87           5 (4Dβ)   47   91           5 (4Dα)   77   79           5 (5Aε)   54   93           5 (5Aδ)   83   78           5 (5Aβ)   103   77           5 (5Aα)   32   93           5 (5Bε)   49   88           5 (5Bδ)   78   103           5 (5Bβ)   74   102           5 (5Bα)   82   105           5 (5Cε)   87   93           5 (5Cδ)   41   103           5 (5Cβ)   72   80           5 (5Cα)   68   62           5 (5Dε)   48   70           5 (5Dδ)   57   96           5 (5Dβ)   76   102           5 (5Dα)   88   78           5 (8Aε)   58   71           5 (8Aδ)   72   89           5 (8Aβ)   103   91           5 (8Aα)   42   83           5 (8Bε)   92   84           5 (8Bδ)   90   112           5 (8Bβ)   50   97           5 (8Bα)   57   80           5 (8Cε)   76   76           5 (8Cδ)   73   72           5 (8Cβ)   111   93           5 (8Cα)   84   65           5 (8Dε)   73   82           5 (8Dδ)   81   63           5 (8Dβ)   68   67           5 (8Dα)   85   66           5 (3Aεκ)           5 (3Aδκ)   66   73           5 (3Aβκ)   64   94           5 (3Aακ)   64   87           5 (3Bεκ)           5 (3Bδκ)   59   93           5 (3Bβκ)   61   95           5 (3Bακ)   100   94           5 (3Cεκ)           5 (3Cδκ)   65   93           5 (3Cβκ)   63   96           5 (3Cακ)   71   99           5 (3Dεκ)   73   88           5 (3Dδκ)   81   97           5 (3Dβκ)           5 (3Dακ)   76   104           5 (4Aεκ)           5 (4Aδκ)   39   102           5 (4Aβκ)   79   70           5 (4Aακ)   43   76           5 (4Bεκ)           5 (4Bδκ)   56   96           5 (4Bβκ)   82   79           5 (4Bακ)   42   77           5 (4Cεκ)           5 (4Cδκ)   52   96           5 (4Cβκ)   67   65           5 (4Cακ)   52   90           5 (4Dεκ)           5 (4Dδκ)   50   83           5 (4Dβκ)   64   97           5 (4Dακ)   80   77           5 (5Aεκ)           5 (5Aδκ)   90   75           5 (5Aβκ)   47   97           5 (5Aακ)   91   103           5 (5Bεκ)           5 (5Bδκ)   46   78           5 (5Bβκ)   63   100           5 (5Bακ)   74   89           5 (5Cεκ)           5 (5Cδκ)   59   99           5 (5Cβκ)   76   92           5 (5Cακ)   93   109           5 (5Dεκ)           5 (5Dδκ)   40   92           5 (5Dβκ)   64   86           5 (5Dακ)   50   86           5 (8Aεκ)   59   92           5 (8Aδκ)   63   97           5 (8Aβκ)   76   96           5 (8Aακ)   53   77           5 (8Bεκ)           5 (8Bδκ)   80   95           5 (8Bβκ)   44   81           5 (8Bακ)   64   103           5 (8Cεκ)   66   86           5 (8Cδκ)   79   95           5 (8Cβκ)   66   95           5 (8Cακ)   31   92           5 (8Dεκ)           5 (8Dδκ)   65   90           5 (8Dβκ)   90   88           5 (8Dακ)   51   84           5 (AG 1M)   89   73           5 (AG 2M)   80   47           5 (AG 3M)   123   63           5 (AG 4M)   100   74           5 (AG 5M)   87   68           5 (AG 6M)   76   57           5 (AG 7M)   99   63           5 (AG 8M)   69   87                      
 
         [0159]    [0159]                                                                       TABLE 3                               5                                                                          % inhibition of                           RNase H       Compound 5   R 2     R 4     R 6     R 8     at 20 μM                                                4.1.7   —CH 2 CH 2 (CH 3 )CH 3     —H   —OCF 3     —H   55       6.1.9   —cC 6 H 12     —H   —CH 3     —H   65       6.1.10   —cC 6 H 12     —H   —SOCH 3     —H   65       4.4.9   —CH 2 CH 2 (CH 3 )CH 3     —CH 2 CH 3     —CH 3     —H   60       5.4.4   —CH 2 (CH 3 )CH 3     —CH 2 CH 3     —CF 3     —H   87       6.4.1   —cC 6 H 12     —CH 2 CH 3     —H   —H   65       6.4.2   —cC 6 H 12     —CH 2 CH 3     —F   —H   75       6.4.3   —cC 6 H 12     —CH 2 CH 3     —OCH 3     —H   60       6.4.4   —cC 6 H 12     —CH 2 CH 3     —CF 3     —H   65       7.4.8   —cCH 2 CH 2 C 6 H 12     —CH 2 CH 3     —H   —F   95       4.5.8   —CH 2 CH 2 (CH 3 )CH 3     —CH 2 CH 2 CH 3     —H   —F   55       7.5.6   —cCH 2 CH 2 C 6 H 12     —CH 2 CH 2 CH 3     —Cl   —H   100       7.5.8   —cCH 2 CH 2 C 6 H 12     —CH 2 CH 2 CH 3     —H   —F   82       1.6.7   —CH 3     —CH 2 CH 2 CH 2 CH 3     —OCF 3     —H   75       2.6.3   —CH 2 CH 2 CH 3     —CH 2 CH 2 CH 2 CH 3     —OCH 3     —H   67       3.6.3   —CH 2 CH 3     —CH 2 CH 2 CH 2 CH 3     —OCH 3     —H   66       3.6.4   —CH 2 CH 3     —CH 2 CH 2 CH 2 CH 3     —CF 3     —H   62       5.6.3   —CH 2 (CH 3 )CH 3     —CH 2 CH 2 CH 2 CH 3     —OCH 3     —H   90       5.6.5   —CH 2 (CH 3 )CH 3     —CH 2 CH 2 CH 2 CH 3     —CH 2 CH 3     —H   100       2.7.3   —CH 2 CH 2 CH 3     —CH 2 CH 2 CH 2 CH 2 CH 3     —OCH 3     —H   60       2.7.10   —CH 2 CH 2 CH 3     —CH 2 CH 2 CH 2 CH 2 CH 3     —SOCH 3     —H   86       3.7.8   —CH 2 CH 3     —CH 2 CH 2 CH 2 CH 2 CH 3     —H   —F   82       3.7.9   —CH 2 CH 3     —CH 2 CH 2 CH 2 CH 2 CH 3     —CH 3     —H   75       3.7.10   —CH 2 CH 3     —CH 2 CH 2 CH 2 CH 2 CH 3     —SOCH 3     —H   85       4.7.2   —CH 2 CH 2 (CH 3 )CH 3     —CH 2 CH 2 CH 2 CH 2 CH 3     —F   —H   58       4.7.3   —CH 2 CH 2 (CH 3 )CH 3     —CH 2 CH 2 CH 2 CH 2 CH 3     —OCH 3     —H   60       4.7.4   —CH 2 CH 2 (CH 3 )CH 3     —CH 2 CH 2 CH 2 CH 2 CH 3     —CF 3     —H   55       4.7.5   —CH 2 CH 2 (CH 3 )CH 3     —CH 2 CH 2 CH 2 CH 2 CH 3     —CH 2 CH 3     —H   100       4.7.10   —CH 2 CH 2 (CH 3 )CH 3     —CH 2 CH 2 CH 2 CH 2 CH 3     —SOCH 3     —H   69       6.7.1   —cC 6 H 12     —CH 2 CH 2 CH 2 CH 2 CH 3     —H   —H   71       6.7.2   —cC 6 H 12     —CH 2 CH 2 CH 2 CH 2 CH 3     —F   —H   76       6.7.3   —cC 6 H 12     —CH 2 CH 2 CH 2 CH 2 CH 3     —OCH 3     —H   65       6.7.4   —cC 6 H 12     —CH 2 CH 2 CH 2 CH 2 CH 3     —CF 3     —H   70       7.7.4   —cCH 2 CH 2 C 6 H 12     —CH 2 CH 2 CH 2 CH 2 CH 3     —CF 3     —H   75       1.8.7   —CH 3     -phenyl   —OCF 3     —H   80       1.8.9   —CH 3     -phenyl   —CH 3     —H   75       1.8.10   —CH 3     -phenyl   —SOCH 3     —H   55       2.8.1   —CH 2 CH 2 CH 3     -phenyl   —H   —H   83       2.8.9   —CH 2 CH 2 CH 3     -phenyl   —CH 3     —H   78       2.8.10   —CH 2 CH 2 CH 3     -phenyl   —SOCH 3     —H   55       3.8.1   —CH 2 CH 3     -phenyl   —H   —H   74       3.8.10   —CH 2 CH 3     -phenyl   —SOCH 3     —H   64       4.8.1   —CH 2 CH 2 (CH 3 )CH 3     -phenyl   —H   —H   80       4.8.4   —CH 2 CH 2 (CH 3 )CH 3     -phenyl   —CF 3     —H   74       4.8.9   —CH 2 CH 2 (CH 3 )CH 3     -phenyl   —CH 3     —H   55       4.8.10   —CH 2 CH 2 (CH 3 )CH 3     -phenyl   —SOCH 3     —H   83       5.8.9   —CH 2 (CH 3 )CH 3     -phenyl   —CH 3     —H   69       5.8.10   —CH 2 (CH 3 )CH 3     -phenyl   —SOCH 3     —H   55       6.8.4   —cC 6 H 12     -phenyl   —CF 3     —H   81       6.8.7   —cC 6 H 12     -phenyl   —OCF 3     —H   87       6.8.10   —cC 6 H 12     -phenyl   —SOCH 3     —H   85       7.8.5   —cCH 2 CH 2 C 6 H 12     -phenyl   —CH 2 CH 3     —H   65       7.8.6   —cCH 2 CH 2 C 6 H 12     -phenyl   —Cl   —H   90       7.8.9   —cCH 2 CH 2 C 6 H 12     -phenyl   —CH 3     —H   61       7.8.10   —cCH 2 CH 2 C 6 H 12     -phenyl   —SOCH 3     —H   68                    
         [0160]    HIV reverse transcriptase is multifunctional, possessing both DNA polymerase and RNase H activities. In separate representative assays, mappicine analogs were tested for their ability to inhibit the RNA-dependent DNA polymerase activity of HIV reverse transcriptase and for their ability to inhibit the RNase H activity of HIV reverse transcriptase. The assay for RNA-dependent DNA polymerase activity is discussed in Borkow, G. et al., “Inhibition of the Ribonuclease H and DNA Polymerase Activities of HIV-1 Reverse Transcriptase by N-(4-tert-Butylbenzoyl)-2-hydroxy-1-naphthaldehyde Hydrazone,”  Biochemistry  1997, 36, 3179-3185, a copy of which is attached hereto and the disclosure of which is incorporated herein by reference. Assays were carried out in the absence and in the presence of mappicine analogs (10 μM final concentration). The results are reported as % residual activity, which is the RNA-dependent DNA polymerase activity of the enzyme in the presence of the mappicine analog divided by the RNA-dependent activity of the enzyme in the absence of the mappicine analog, multiplied by 100.  
         [0161]    The assay for RNase H activity of HIV reverse transcriptase was the fluorescence-based assay described in detail in the Experimental section and discussed in U.S. Provisional Patent Application Serial No. 60/318,359. Assays were carried out in the absence and in the presence of mappicine analogs (10 μM or 20 μM final concentration). The results are reported as % residual activity, which is the RNase H activity of the enzyme in the presence of the mappicine analog divided by the RNase H activity of the enzyme in the absence of the mappicine analog, multiplied by 100. Alternatively, the results are reported as % inhibition, which is calculated as the ratio of the RNase H activity of the enzyme in the presence of the mappicine analog to the enzyme activity in the absence of the mappicine analog, multiplied by 100, and then subtracting this number from 100.  
         [0162]    The results of biological testing of the mappicine analogs of the present invention were quite surprising. At a concentration of 10 μM or 20 μM , the mappicine analogs tested showed inhibitory activity against HIV reverse transcriptase via inhibitory activity against the RNA-dependent DNA polymerase activity and/or against the RNase H activity of HIV reverse transcriptase. In general, mappicine analogs that very closely resembled the natural product were less active than more distant analogs, differing, for example, in at least two substituents. Some of these more distant analogs were, however, found to be quite active. While many mappicine analogs tested surprisingly exhibited an appreciable level of RNase H inhibition at the concentration level of the studies, mappicine ketone analogs (illustrated in formula (2) above, and previously shown to be active against DNA viruses) were somewhat less active than other mappicine analogs (for example, mappicine alcohols, in which R 1  of formula (1) above is H).  
         [0163]    Two quite active mappicine analogs of the present invention, 5-AG 2M and 5-AG 6M, are shown below. Dose-response curves for mappicine alcohols 5-AG 2M and 5-AG 6M are illustrated in FIGS. 11 and 12, respectively. Both of these mappicine possess inhibitory activity against HIV-1 RNase H (IC 50 &lt;10 μM; see Table 4) comparable to the known RNase H inhibitor BBNH. The results have been confirmed in cell culture viral growth assays (Table 4), thereby supporting the postulate that RNase H binding and anti-viral activity are linked.  
                         
 
         [0164]    Based on the above results, mappicine analogs exhibit strong potential to provide extremely potent RNase H inhibitors. Such potent inhibitors are a welcome addition to the current arsenal for treatment of AIDs or other retroviral diseases, either alone or in combination with existing drugs.  
                                     TABLE 4                           IC 50  (μM) against HIV RNase H   EC 50  (μM) against       COMPOUND   in vitro   HIV-1 replication                                5-AG 2M   8   ≈5       5-AG 6M   10   ≈5       BBNH   1.8   1.5                  
 
         [0165]    Other very active mappicine analogs include the 5{4.5.7} and 5{7.5.6} analogs. The structure of analog 5{7.5.6} is shown below. Dose-response curves for mappicine alcohols 5{4.5.7} and 5{7.5.6} are illustrated in FIG. 13. Both of these mappicine possess inhibitory activity against HIV-1 RNase H. Antiviral and toxicity data for 5{7.5.6} is set forth in FIG. 14.  
         [0166]    Other than the mappicine analogs described herein, there have been only three published reports of compounds able to inhibit HW-1 reverse transcriptase associated RNase H activity with IC 50 &lt;10 μM. These compounds are N-4-(t-butylbenzoyl)-2-hydroxy-1-naphthaldehyde hydrazone (BBNH, see for example Borkow, G. et al., “Inhibition of the Ribonuclease H and DNA Polymerase Activities of HIV-1 Reverse Transcriptase by N-4-(tert-butylbenzoyl)-2-hydroxy-1-naphthaldehyde hydrazone,”  Biochemistry  1997, 36, 3179-3185), 2-[(4-chlorophenyl)-hydrazono]-malonic acid (CPHM, see for example Gabbara, S. et al., “Inhibitors of DNA Strand Transfer Reactions Catalyzed by HIV-1 Reverse Transcriptase,”  Biochemistry  1999, 38, 13070-13076), and 4-[5-(benzoylamino)thien-2-yl]-2,4-dioxobutanoic acid (BTOBA, see for example Shaw-Reid, C. et al., “Inhibition of HIV-1 ribonuclease H by a novel diketo acid, 4-[5-(benzoylamino)thien-2-yl]-2,4-dioxobutanoic acid,”  Journal of Biological Chemistry  2003, 278, 2777-2780). Of these three, only BBNH has demonstrated antiviral activity against HIV-1 replication in cultured cells. However, BBNH is highly toxic to cells at concentrations only slightly above those that inhibit virus replication. Neither CPHM nor BTOBA are able to penetrate cells, and thus they cannot inhibit HIV-1 replication in cultured cells. Thus, BBNH, CPHM and BTOBA do not have therapeutic potential for the treatment of HIV-1 infection in humans. In contrast to BBNH, CPHM and BTOBA, the mappicine analogs described herein are capable of inhibiting HIV-1 replication in cultured cells and show little toxicity to cells. Thus, they provide the first example of an RNase H inhibitor with potential therapeutic utility.  
         [0167]    Moreover, the mappicine analogs of the present invention may provide potent inhibitors and methods of inhibition and/or treatment of even higly resistant strains of HIV-1-. For example, mappicine 5{7.5.6} is a potent inhibitor of the replication of wild-type 1HV-1 in cultured cells, inhibiting this replication with an EC 50  of approximately 3 μM. Mappicine 5{7.5.6} also potently inhibits the replication of two mutant HIV-1 strains with high-level resistance to nevirapine, delavirdine and efavirenz, the three normucleoside reverse transcriptase inhibitors (NNRTI) approved for clinical treatment of HIV-1 infection. These two strains possess the mutations K103N+Y181C or V106A+Y181C in the viral reverse transcriptase, which result in more than 1000-fold resistance to the clinically used NNRTI. Mappicine 5{7.5.6} inhibited the replication of these two resistant virus strains with EC 50  values of 3.5 and 4.2 μM, respectively.  
         [0168]    Pharmaceutical Compositions  
         [0169]    The present invention provides a broad variety of compositions prepared from compounds of the present invention. Such compositions have utility for human and veterinary antiviral use, and for treating viral infections in plants, e.g., agricultural or ornamental seeds and plants. Such compositions comprise a carrier which is acceptable for the intended end use together with at least one inventive compound. For example, in veterinary use, the carrier may be a liquid, or spray, or may be formulated in a solid, non-degradable or degradable form for insertion in the rumen. For agricultural use, the compound can be mixed with a fertilizer, other microbiocides such as fungicides, or insecticides and the like. The present compounds may also be formulated in powders or sprays for application to plant surfaces.  
         [0170]    The pharmaceutical compositions of this invention comprise one or more compounds of the present invention in admixture with an inert pharmaceutically acceptable carrier or diluent. Compositions may contain an effective amount of the inventive compound in one unit, such as in a single pill, capsule, or pre-measured intravenous dose or pre-filled syringe for injection, or, as is frequently the case, the composition may be prepared in individual dose forms where one unit, such as a pill, contains a sub-optimal dose with the user being instructed to take two or more unit doses per treatment. When the composition is presented as a cream, it contains a discrete amount of drug and the user applies an effective amount of the cream one or more times until the disease is in remission or has been effectively treated. Concentrates for later dilution by the end user may also be prepared, for instance for IV formulations and multi-dose injectable formulations.  
         [0171]    Carriers or diluents contemplated for use in these compositions are generally known in the pharmaceutical formulary arts. Reference to useful materials can be found in well known compilations such as Remington&#39;s Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa., 18042, U.S.A.  
         [0172]    The nature of the composition and the pharmaceutically acceptable carrier or diluent will, of course, depend upon the intended route of administration, for example, by intravenous and intramuscular injection, parenterally, topically, orally, or by inhalation.  
         [0173]    For parenteral administration the pharmaceutical composition may be in the form of a sterile injectable liquid such as an ampule or an aqueous or nonaqueous liquid suspension.  
         [0174]    For topical administration the pharmaceutical composition may be in the form of a cream, ointment, liniment, lotion, paste, spray or drops suitable for administration to the skin, eye, ear, nose or genitalia.  
         [0175]    For oral administration the pharmaceutical composition may be in the form of a tablet, capsule, powder, pellet, atroche, lozenge, syrup, liquid, or emulsion.  
         [0176]    The pharmaceutically acceptable carrier employed may be either a solid or liquid. Exemplary of solid carriers are lactose, kaolin, terra alba, sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate, mannitol, stearic acid and the like.  
         [0177]    Examples of appropriate pharmaceutically acceptable liquid carriers or diluents include: for aqueous systems, water; for non-aqueous systems, ethanol, glycerin, propylene glycol, corn oil, cottonseed oil, peanut oil, sesame oil, liquid paraffins and mixtures thereof with water. For aerosol systems, pharmaceutically acceptable carriers include dichlorodifluoromethane, chlorotrifluoroethylene and compressed carbon dioxide. Also, in addition to the pharmaceutical carrier or diluent, the instant compositions may include other ingredients such as stabilizers, antioxidants, preservatives, lubricants, suspending agents, viscosity modifiers and the like, provided that the additional ingredients do not have a detrimental effect on the therapeutic action of the instant compositions. Similarly, the carrier or diluent may include time delay materials well known to the art, such as glyceryl monostearate or glyceryl distearate alone or with a wax, ethylcellulose, hydroxypropylmethylcellulose, methylmethacrylate and the like.  
         [0178]    To obtain a stable water soluble dose form, a pharmaceutically acceptable salt of a compound of the present invention is dissolved in an aqueous solution of an organic or inorganic acid or base. If a soluble salt form is not available, the inventive compound may be dissolved in a suitable co-solvent or combinations thereof. Examples of such suitable cosolvents include, but are not limited to, alcohol, propylene glycol, polyethylene glycol 300, polysorbate 80, glycerin and the like in concentrations ranging from 0-60% of the total volume.  
         [0179]    It will be appreciated that the actual preferred dosages of the compounds of the present invention used in the pharmaceutical and other compositions of this invention will vary according to the particular complex being used, the particular composition formulated, the mode of administration and the particular site, host and disease being treated. These compounds are active in the concentration ranges of two commercial antiviral drugs, Cytovene (ganciclovir) and Zovirax (acyclovir). For example, the latter is manufactured in 200 mg capsules with instructions for treating herpes simplex viruses by taking one capsule every 4 hours, but not to exceed 5 capsules per day.  
       EXAMPLES  
       [0180]    Assay for Ribonuclease H Activity Based on Fluorescence Resonance Energy Transfer (FRET)  
         [0181]    Preparation of the Fluorophore-RNA/Quencher-DNA Hybrid Duplex Substrate.  
         [0182]    The RNA/DNA hybrid duplex substrate for use in the assay comprises an RNA oligonucleotide of sequence 5′-GAU CUG AGC CUG GGA GCU-3′, modified at the 3′-end with Aminolink-2 and derivatized with fluorescein isothiocyanate, to provide a modified RNA oligonucleotide of the sequence 5′-GAU CUG AGC CUG GGA GCU-fluorescein-3′, annealed to a complementary DNA oligonucleotide of the sequence 5′-AGC TCC CAG GCT CAG ATC-3′ modified at the 5′-end with Aminolink-2 and derivatized with the FRET acceptor DABCYL succinimidyl ester, to provide a modified DNA oligonucleotide of the sequence 5′-DABCYL-AGC TCC CAG GCT CAG ATC-3′.  
         [0183]    To prepare the 3′-fluorescein-RNA/5′-DABCYL-DNA hybrid duplex substrate, a known amount of 3′-fluorescein-RNA was dissolved in 20 mM Tris buffer (pH 8.0, 37° C.) to provide a final concentration of 5 μM. Two equivalents of the 5′-DABCYL-DNA oligonucleotide were added, and the mixture was heated to 90° C. for 5 min and cooled slowly to room temperature to allow duplex formation. The positioning of the fluorescein donor at the 3′-end of the RNA oligonucleotide and the DABCYL acceptor at the 5′-end of the DNA oligonucleotide provides a very close proximity of the donor and acceptor that results in a very intense quenching of the fluorescein emission in the intact RNA/DNA hybrid duplex substrate due to the spectral overlap of the fluorescence emission of fluorescein with the absorption spectrum of DABCYL. In addition, DABCYL is non-fluorescent, and thus cannot contribute any light emission. Both of these factors result in a very low background and provide a high signal-to-noise in the assay measurements. The ratio of the donor fluorescence in the absence and in the presence of its quencher is approximately fifteen-fold.  
         [0184]    Microplate Assay Protocol for the Measurement of RNase H Activity Using the RNA/IDNA Hybrid Duplex Substrate.  
         [0185]    Reaction assay mixtures contained 5 μl of a stock solution of 2.5 μM RNA/DNA hybrid duplex substrate added to 85 μl of assay buffer (50 mM Tris, pH 8.0, 37° C., containing 60 mM KCl and 2.5 mM MgCl 2 ), prepared in the wells of a 96-well fluorescence microtiter plate, and warmed to 37° C. using the temperature control of the SpectraMax Gemini XS microplate spectrofluorometer (Molecular Devices). Reactions were started by the addition of 5 μl of a solution of recombinant HIV-1 reverse transcriptase (usually providing a final concentration of 2.5 nM of the p51/p66 RT heterodimer in the assay), and mixing using the automatic mixing function of the microplate spectrofluorometer. The increase in fluorescence signal resulting from the loss of FRET due to the enzymatic hydrolysis of the RNA strand was measured over suitable time intervals (ranging from 3 minutes to 60 minutes), at an excitation wavelength of 490 nm and an emission wavelength of 528 nm, using a cut-off filter of 515 nm. Data analysis and curve fitting were carried out using the appropriate transform functions of the software SigmaPlot 2000 (SPSS Inc.).  
         [0186]    Synthesis  
         [0187]    4-(1-Hydroxypropyl)-6-iodo-3-methyl-1-pent-2-ynyl-1H-pyridin-2-one (3a).  
         [0188]    To a solution of 4-(1-hydroxypropyl)-6-iodo-3-methyl-1H-pyridin-2-one (1) (prepared as set forth in de Frutos, O.; Curran, D. P.  J. Comb. Chem.  2000, 2, 639) (123 mg, 0.42 mmol) in DME (4.0 mL) and DMF (1.0 mL) at 0° C. was added portionwise NaH (20 mg, 0.505 mmol, 60% in mineral oil). After 15 min, LiBr (124 mg, 0.84 mmol) was added and the cooling bath removed. 2-Pentynyl bromide (73 mg, 0.84 mmol) was added 15 min later and the mixture was heated in the dark at 65° C. for 20 h. After cooling, the reaction was diluted with EtOAc, washed with brine, dried over Na 2 SO 4  and evaporated. The residue was purified by flash chromatography (gradient CH 2 Cl 2  to CH 2 Cl 2 /EtOAc 80:20) to yield 3a (99 mg, 66%) as a white foam:  1 H NMR (300 MHz, CDCl 3 ) δ 0.91 (t, J=7.3 Hz, 3H), 1.1 (t, J=7.4 Hz, 3H), 1.53-1.67 (m, 2H), 1.89 (s, 3H), 2.16 (q, J=7.4 Hz, 2H), 3.49 (bs, 1H), 4.63 (t, J=6.5 Hz, 1H), 4.94 (d, J=16.8 Hz, 1H), 5.02 (d, J=16.8 Hz, 1H), 7.07 (s, 1H);  13 C NMR (75 MHz, CDCl 3 ). δ 10.09, 12.14, 12.59, 13.58, 29.89, 44.75, 70.37, 73.20, 86.75, 94.95, 118.64, 124.04, 153.29, 162.62; IR (film, NaCl, cm-1) 3398, 2977, 1629, 1519, 1177; LRMS (70 eV, El) m/z (rel int %) 359 (M+), 344 (100), 254, 128, 93, 77, 67, 59; HRMS m/z calcd for C 14 H 18 NO 2 I (M+) 359.0382, found 359.0393.  
         [0189]    General Procedure for Cascade Radical Reaction.  
         [0190]    To a solution of iodopyridone (15 mg) in benzene (0.5 mL) was added the corresponding isonitrile (3.0 equiv) and hexamethylditin (2 equiv). The mixture was irradiated at room temperature with a 275W GE sunlamp for 4 h and 30 min. The solvent was evaporated and the residue purified by flash chromatography.  
         [0191]    2-tert-Butyloxycarbonylamino-7-(1-hydroxypropyl)-8-methyl-12-trimethylsilanyl-11H-indolizino [1,2-b]quinolin-9-one (5-AG-2M).  
         [0192]    Treatment of 4-(1-hydroxypropyl)-6-iodo-3-methyl-1-[3-(trimethylsilanyl)prop-2-ynyl]-1H-pyridin-2-one (3b) (15.0 mg, 0.037 mmol) according to the cascade radical reaction general (see, de Frutos, O.; Curran, D. P.  J. Comb. Chem.  2000, 2, 639) procedure afforded 5-AG-2M (12.3 mg, 67%) as a pale brown solid, after purification of the crude residue by flash chromatography (gradient CH 2 Cl 2  to CH 2 Cl 2 /acetone 1:1):  1 H NMR (300 MHz, CDCl 3 ) δ 0.69 (s, 9H), 0.97 (t, J=7.3 Hz, 3H), 1.61 (s, 9H), 1.78-1.88 (m, 2H), 2.16 (s, 3H), 3.89 (bs, 1H), 4.87 (t, J=6.9 Hz, 1H), 5.09 (d, J=18.8 Hz, 1H), 6.73 (s, 1H), 7.19 (d, J=7.0 Hz, 1H), 7.68 (d, J=7.0 Hz, 1H), 8.29 (s, 1H); LRMS (70 eV, El) m/z (rel int %) 493 (M+), 437 (100), 419, 404, 393, 378, 73; HRMS m/z calcd for C 27 H 35 N 3 O 4 Si (M+) 493.2397, found 493.2388.  
         [0193]    2-Amino-7-(1-hydroxypropyl)-8-methyl-12-trimethylsilanyl-1 1H-indolizino[1,2-b]quinolin-9-one (5-AG-3M).  
         [0194]    To a solution of 5-AG-2M (9.8 mg, 0.02 mmol) in CH 2 Cl 2  (0.5 mL) was added TFA (0.3 mL). The mixture was stirred at room temperature for 5 h and then concentrated under reduced pressure. The residue was purified by flash chromatography (gradient CH 2 Cl 2  to CH 2 Cl 2 /acetone 1:1) to yield 5-AG-3M (5.1 mg, 65%) as an orange solid:  1 H NMR (300 MHz, CD 3 OD) δ 0.63 (s, 9H), 1.02 (t, J=7.3 Hz, 3H), 1.71-1.77 (m, 2H), 2.13-2.20 (m, 3H), 5.28 (s, 2H), 7.29 (dd, J=2.1 and 8.9 Hz, 1H), 7.39 (d, J=2.1 Hz, 1H), 7.54 (s, 1H), 7.88 (d, J=8.9 Hz, 1H;  13 C NMR (125 MHz, CDCl 3 ) δ 1.86, 10.21, 12.15, 29.99, 51.87, 71.14, 99.32, 124.86, 126.51, 127.89, 128.88, 130.31, 131.29, 134.61, 142.14, 143.46, 147.00, 151.01, 154.32, 161.39; LRMS (70 eV, EI) m/z (rel int %) 393 (M+), 378, 364, 73, 57; HRMS m/z calcd for C 22 H 27 N 3 O 2 Si (M+) 393.1872, found 393.1883.  
         [0195]    [0195] 2 -tert-Butyloxycarbonylamino-12-ethyl-7-(1-hydroxypropyl)-8-methyl-11H-indolizino[1,2-b]quinolin-9-one (5-AG-5M).  
         [0196]    Treatment of 3a (15.0 mg, 0.04 mmol) according to the cascade radical reaction general procedure afforded 5-AG-5M (13.2 mg, 70%) as a brown solid, after purification of the crude residue by flash chromatography (gradient CH 2 Cl 2  to CH 2 Cl 2 /acetone 1:1):  1 H NMR (300 MHz, CD 3 OD) δ 0.97 (t, J=7.3 Hz, 3H), 1.34 (t, J=7.5 Hz, 3H), 1.63 (s, 9H), 1.79-1.87 (m, 2H), 2.18 (s, 3H), 2.64 (s, 1H), 2.91-3.09 (m, 2H), 4.87 (t, J=6.9 Hz, 1H), 4.96 (d, J=18.5 Hz, 1H), 5.17 (d, J=18.5 Hz, 1H), 7.32 (d, J=9.0 Hz, 1H), 7.74 (d, J=9.1 Hz, 1H), 7.88 (s, 1H); LRMS (70 eV, E1) m/z (rel int %) 449 (M+), 393, 375, 358, 349 (100), 332, 320, 91, 57; HRMS m/z calcd for C 26 H 31 N 3 O 4  (M+) 449.2314, found 449.2312.  
         [0197]    2-Amino-12-ethyl-7-(1-hydroxypropyl)-8-methyl-1 1H-indolizinol[1,2-b]quinolin-9-one (5-AG-6M).  
         [0198]    To a solution of 5-AG-5M (10.4 mg, 0.023 mmol) in CH 2 Cl 2  (0.5 mL) was added TFA (0.3 mL). The mixture was stirred at room temperature for 5 h and then concentrated under reduced pressure. The residue was purified by flash chromatography (gradient CH 2 Cl 2  to CH 2 Cl 2 /acetone 1:1) to yield 5-AG-6M (6.4 mg, 79%) as an orange solid:  1 H NMR (300 MHz, CD 3 OD) δ 1.02 (t, J=7.4 Hz, 3H), 1.35 (t, J=7.5 Hz, 3H), 1.71-1.89 (m, 2H), 2.20 (s, 3H), 3.11 (q, J=7.5 Hz, 2H), 5.14 (s, 2H), 7.21 (d, J=2.2 Hz, 1H), 7.30 (dd, J=2.2 and 9.1 Hz, 1H), 7.53 (s, 1H), 7.84 (d, J=9.1 Hz, 1H);  13 C NMR (125 MHz, CD 3 OD) δ 10.58, 12.27, 13.9, 23.94, 31.36, 50.89, 72.23, 104.58, 124.77, 128.93, 130.29, 130.6, 143.6, 144.28, 145.21, 148.69, 156.91, 163.32; LRMS (70 eV, EI) m/z (rel int %) 349 (M+), 322, 101, 91, 81, 69, 57 (100); HRMS m/z calcd for C 21 H 23 N 3 O 2  (M+) 349.1790, found 349.1800.  
         [0199]    2-Acetoxy-12-ethyl-7-(1-hydroxypropyl)-8-methyl-1H-indolizino [1,2-b]quinolin-9-one (5-AG-7M).  
         [0200]    Treatment of 3a (15.0 mg, 0.04 mmol) according to the cascade radical reaction general procedure afforded 5-AG-7M (16.1 mg, 98%) as a pale brown solid, after purification of the crude residue by flash chromatography (gradient CH 2 Cl 2  to CH 2 Cl 2 /acetone 1:1):  1 H NMR (300 MHz, CDCl 3 ) δ 0.98 (t, J=7.4 Hz, 3H), 1.26 (t, J=7.6 Hz, 3H), 1.65-1.72 (m, 1H), 1.79-1.89 (m, 1H), 2.17 (s, 3H), 2.41 (s, 3H), 277 (q, J=7.6 Hz, 2H), 4.86-4.97 (m, 3H), 5.17 (d, J=18.6 Hz, 1H), 7.35-7.41 (m, 3H), 7.86 (d, J=8.9 Hz, 1H)); LRMS (70 eV, EI) m/z (rel int %) 392 (M+), 350, 333, 167, 149 (100), 129, 99, 91, 71, 59; HRMS m/z calcd for C 23 H 24 N 2 O 3  (M+) 392.1736, found 392.1744.  
         [0201]    12-Ethyl-2-hydroxy-7-(1-hydroxypropyl)-8-methyl-11H-indolizino[1,2-b]quinolin-9-one (5-AG-8M).  
         [0202]    To a solution of 5-AG-7M (13 mg, 0.033 mmol) in MeOH (0.4 mL) and H 2 O (0.4 mL) was added K 2 CO 3  (13.0 mg, 0.09 mmol). The mixture was stirred at room temperature for 2 h and then concentrated under reduced pressure. The residue was purified by flash chromatography (gradient CH 2 Cl 2  to CH 2 Cl 2 /acetone 1:1) to yield 5-AG-8M (9.1 mg, 78%) as a pale solid:  1 H NMR (300 MHz, CD 3 OD) δ 1.02 (t, J=7.3 Hz, 3H), 1.38 (t, J=7.5 Hz, 3H), 1.72-1.80 (m, 2H), 2.22 (s, 3H), 3.10-3.17 (m, 2H), 5.21 (s, 2H), 7.35-7.38 (m, 2H), 7.57 (s, 1H), 7.98 (d, J=9.8 Hz, 1H));  13 C NMR (125 MHz, CD 3 OD) δ 10.58, 12.26, 14.03, 22.5, 31.38, 72.24, 100.99, 106.19, 119.32, 123.76, 124.94, 128.85, 129.93, 131.91, 144.63, 145.17, 145.61, 150.82, 156.94, 158.46, 163.40; LRMS (70 eV, E1) m/z (rel int %) 350 (M+, 100), 333, 317, 292, 166, 69; HRMS m/z calcd for C 21 H 22 N 2 O 3  (M+) 350.1630, found 350.1632.  
         [0203]    Fluorous Mixture Synthesis of the Mappicine Library  
         [0204]    General LC-MS analysis conditions: Fluofix colum (4.6×250 mm, Keystone Scientific, Inc.), gradient 90% MeOH—H 20  to 100% MeOH in 15 min, then maintain 100% MeOH for 5-10 min. Mass spectrometer detection with positive APCI ionization source. Similar conditions were applied to F-HPLC analyses of intermediates in the mixture synthesis.  
         [0205]    Modified procedure for the preparation of 18. To a solution of aldehyde 16 (3.69 g, 111.0 mmol) in ethanol (20 mL) at −40° C. was added NaBH4 (419 mg, 11.0 mmol). The reaction mixture was further stirred for 1 h at −40° C. and then quenched with water. The crude product was purified by silica gel column chromatography (10% EtOAc/hexanes) to give alcohol 17 (2.60 g, 70%) as a colorless oil. To a solution of alcohol 17 (18.40 g, 54.44 mmol) in 1,2-dichloroethane (75 mL) was added triethylsilane (63.15 g, 0.54 mol) followed by slow addition of boron trifluoride etherate (34.5 ml, 0.27 mmol) at room temperature. The reaction mixture was then heated at 75° C. for 2 h before quenching with aq. NaHCO 3 . After extraction with diethyl ether, the organic layer was dried and passed through a silica plug with hexanes to give the pure product 18 (15.8 g, 90%).  
         [0206]    General procedure for tagging alcohols 21{1-7} with perfluoroalkylsilanes (RfCH 2 CH 2 (iPr) 2 SiH). Preparation of 9{3}. To C 6 F 13 CH 2 CH 2 (iPr) 2 SiH (9.88 g, 21.4 mmol) was added trifluoromethanesulfonic acid (2.05 mL, 16.4 mmol) at 0° C. The reaction mixture was then stirred at room temperature for 15 h. A solution of 21{3} (4.16 g, 16.4 mmol) and 2,6-lutidine (3.8 mL, 32.8 mmol) in dry CH 2 Cl 2  (40 mL) was added. After stirring at room temperature for 2 h, the reaction mixture was quenched with aq. NH 4 Cl and extracted with CH 2 Cl 2  and ether. The combined organic layers were dried and chromatography on silica gel with EtOAc/hexanes (5/95) gave 9{3} (9.97 g, 85%) as a colorless clear oil.  1 H NMR (300 MHz, CDCl 3 ) □ 0.25 (s, 9H), 0.77 (dd, 2H), 0.88 (t, 3H), 0.97-1.08 (m, 14H), 1.67 (m, 2H), 1.72-1.98 (m, 2H), 2.10 (s, 3H), 4.91 (s, 1H), 7.21 (s, 1H).  
         [0207]    General procedure for TMS-Iodo exchange. Preparation of 10{1-7}. A mixture of seven tagged alcohols 9{1-7} (1.5 mmol each, total 10.5 mmol, 8.2 g,) in CH 2 Cl 2 /CCl 4  (3:1, 80 mL ) was sonicated at 15° C. To this mixture was added a solution of ICl (5.1 g, 31.5 mmol) in CH 2 Cl 2 /CCl 4  (3:1, 55 mL) over 30 min via an addition funnel. The reaction was followed by F-HPLC analysis for completion. The mixture was washed with aq. Na 2 S 2 O 3 , the aqueous layer was extracted with ether, and the organic layer was concentrated to give 10{1-7} (8.9 g) as a clear yellow oil. This crude product was used for the next reaction without further purification.  
         [0208]    General procedure for demethylation. Preparation of 11{1-7}. To a solution of 10{1-7} (8.7 g crude) in CHCl 3  (150 mL) was added BBr 3  (7.9 g, 31.5 mmol) at room temperature. The reaction mixture was then refluxed for 2.5 h. The reaction was followed by F-HPLC analysis for completion. The cooled reaction mixture was slowly poured into aq. NaHCO 3 . The organic layer was separated, the aqueous layer was extracted with ether, and the combined organic layers were washed with brine and concentrated. The crude product was purified by chromatography on silica gel with hexanes followed by hexanes/AcOEt (9:1) gave 11{1-7} (6.50 g, 76% from 10{1-7}) as a clear brown oil. The purity was checked by F-HPLC.  
         [0209]    General procedure for N-propargylation. To a mixture of seven pyridones 11{1-7} (0.30 mmol each, total 2.1 mmol) in DME (6 mL) and DMF (2 mL) was added NaH (60% in mineral oil, 0.095 g, 2.40 mmol) at 0° C. followed by LiBr (0.37 g, 4.2 mmol) after 10 min. The reaction mixture was warmed up to room temperature. Propargyl bromide 19{1-8} (3.1 mmol) was added. The mixture was heated at 75° C. for 7 h. The cooled reaction mixture was extracted with ether and washed with brine. The concentrated organic layer was purified by chromatography on silica gel with hexanes followed by hexanes/AcOEt (8:2) to give 12{1-7,1-8} in an average yield of 70%. The purity was checked by F-HPLC.  
         [0210]    General procedure for radical annulation. To a mixture of seven N-propargyl pyridones 12{1-7,1-8} (0.024 mmol each, total 0.17 mmol) was added hexamethyl ditin (15 μL, 0.069 mmol) and a solution of aryl isonitrile (1.0 M in benzene, 0.5 ml, 0.5 mmol). The mixture was purged with nitrogen for 5 min and sealed in a vial. The mixture was irradiated with a sunlamp for 5 h and then loaded onto a SPE cartridge packed with 2.5 g silica. The cartridge was eluted with 10% EtOAc/hexanes (20 mL) followed by 15% MeOH/EtOAc (10 mL). The MeOH/EtOAc fraction was evaporated to dryness to give a mixture of seven tagged mappicines 13{1-7,1-8,1-10}. All eighty mixtures were analyzed by LC-MS before the next step of demixing.  
         [0211]    General procedure for demixing of tagged mappicines 13{1-7,1-8,1-10}. Demixing of 80 tagged mappicines mixtures was carried out on a Waters HPLC system (manually) or a Gilson serial HPLC with an automatic fraction collector. The separation conditions were as follows: Fluophase-RP column (20×250 mm, 5 μm, Keystone Scientific, Inc.), gradient 88% MeOH-12% H 2 O to 100% MeOH in 28 min, then to 100% THF in 7 min with a flow rate of 12 mL/min. A mixture sample dissolved in a minimum amount of THF was injected. The demixed fractions were collected by following the UV detector signal (manually) or by an automatic fraction collector. The desired fractions were concentrated to give 560 individually pure tagged mappicines 13{1-7,1-8,1-10} (3-6 mg each).  
         [0212]    General procedure for detagging. Preparation of individually pure mappicines 5{1-7,1-8,1-10}. Detagging of 560 13{1-7,1-8,1-10} was accomplished in parallel. To each vial containing a tagged mappicine (3-6 mg) in THF (0.2 mL) was added 10 drops of HF-pyridine at room temperature and then heated at 60° C. The reactions were followed by TLC for completion. Reactions of shorter-tagged (C 4 F 9  to C 6 F 13 ) mappicine analogs took 1 h to complete whereas longer reaction times (up to 10 h) for long-tagged (C 7 F 15  to C 10 F 21 ) mappicine analogs were necessary for the reaction to go to completion. Upon completion of the reaction, the mixture was diluted with EtOAc, washed with aq. NaHCO 3  and the organic layers air-dried. Upon solvent evaporation, the residue from each vial was subjected to solid-phase extraction on reverse-phase silica gel (0.5 g) packed into syringe cartridges of 2.5 ml volume. The residue was dissolved in a minimum amount of 80:20 MeOH:H 2 O (several drops of THF were sometimes added to aid dissolve the residue) and loaded onto the pre-wet (80:20 MeOH:H 2 O)SPE cartridge which was set on a 12×2 SPE manifold. The first fraction (5 to 8 mL) eluted with 80:20 MeOH:H 2 O was collected, transferred into a vial, and air-dried giving the mappicine analog in an average amount of 1-2 mg. The purity was assessed by HPLC analysis (Novapak C 18  column, MeOH—H 2 O gradient) of 20% of randomly selected library samples. Additional structure characterizations including MS, LC-MS,  1 H NMR, and LC-NMR analyses were also carried out.  
         [0213]    Table 5 below sets forth liquid chromatography-mass spectrometry (LC-MS) data of eight mixtures of seven N-propargylation products 12{1-7,1-8}. Table 6 sets forth LC-MS data of eighty mixtures of seven tagged-mappicines 13{1-7,1-8,1-10}, and Table 7 sets forth MS data of selected mappicine analogs 5.  
                                                                                   TABLE 5                       MS with positive APCI                                12{1-7, 1}   {1, 1}   {2, 1}   {3, 1}   {4, 1}   {5, 1}   {6, 1}   {7, 1}                    RT (min)   3.4   4.5   6.7   8.9   11.0   12.4   15.6       MS   628   706   792   870   906   996   1074               12{1-7, 2}   {1, 2}   {2, 2}   {3, 2}   {4, 2}   {5, 2}   {6, 2}   {7, 2}               RT (min)   3.2   4.3   6.3   8.3   10.3   11.7   14.9       MS   734   812   898   976   1012   1102   1180               12{1-7, 3}   {1, 3}   {2, 3}   {3, 3}   {4, 3}   {5, 3}   {6, 3}   {7, 3}               RT (min)   3.5   4.7   6.9   9.1   11.2   12.6   15.8       MS   642   720   806   884   920   1010   1088               12{1-7, 4}   {1, 4}   {2, 4}   {3, 4}   {4, 4}   {5, 4}   {6, 4}   {7, 4}               RT (min)   3.6   4.9   7.3   9.5   11.7   13.0   16.1       MS   656   734   820   898   934   1025   1102               12{1-7, 5}   {1, 5}   {2, 5}   {3, 5}   {4, 5}   {5, 5}   {6, 5}   {7, 6}               RT (min)   3.8   5.2   7.7   10.0   12.2   13.5   16.5       MS   670   748   834   912   948   1038   1117               12{1-7, 6}   {1, 6}   {2, 6}   {3, 6}   {4, 6}   {5, 6}   {6, 6}   {7, 6}               RT (min)   4.0   5.5   8.1   10.3   12.6   13.8   16.8       MS   684   762   848   926   962   1052   1131               12{1-7, 7}   {1, 7}   {2, 7}   {3, 7}   {4, 7}   {5, 7}   {6, 7}   {7, 7}               RT (min)   4.2   5.7   8.4   10.7   12.9   14.2   17.1       MS   698   777   863   940   976   1066   1144               12{1-7, 8}   {1, 8}   {2, 8}   {3, 8}   {4, 8}   {5, 8}   {6, 8}   {7, 8}               RT (min)   3.2   4.4   6.5   8.5   10.6   12.0   15.2       MS   704   782   868   946   982   1073   1150                  
 
         [0214]    [0214]                                                                                                                                                                                                                                                                                                                                         TABLE 6                           13{1-7, 1, 1-10}, MS with positive APCI            13{x, 1, y}   x = {1}   {2}   {3}   {4}   {5}   {6}   {7}               Y = {1}   603   681   767   845   881   971   1049       {2}   621   699   785   863   899   989   1067       {3}   633   711   797   875   911   1001   1079       {4}   671   749   835   913   949   1039   1117       {5}   631   709   795   873   909   999   1077       {6}   637   715   801   879   915   1005   1083       {7}   687   765   851   929   965   1055   1133       {8}   621   699   785   863   899   989   1067       {9}   617   695   781   859   895   985   1063       {10}    649   727   813   891   927   1017   1095                    13{1-7, 2, 1-10}, MS with positive APCI            13{x, 2, y}   x = {1}   {2}   {3}   {4}   {5}   {6}   {7}               Y = {1}   719   787   873   953   987   1077   1155       {2}   727   805   891   969   1005   1095   1173       {3}   739   817   903   981   1017   1107   1185       {4}   777   855   941   1019    1055   1145   1223       {5}   737   815   901   979   1015   1105   1183       {6}   743   821   907   985   1021   1111   1189       {7}   793   871   957   1035    1071   1161   1239       {8}   727   805   891   909   1005   1095   1173       {9}   723   801   887   965   1001   1091   1169       {10}    755   833   919   997   1033   1123   1201                    13{1-7, 3, 1-10}, MS with positive APCI            13{x, 3, y}   x = {1}   {2}   {3}   {4}   {5}   {6}   {7}               Y = {1}   617   695   781   859   895   985   1063       {2}   635   713   799   877   913   1003   1081       {3}   647   725   811   889   925   1015   1093       {4}   685   763   849   927   963   1053   1131       {5}   645   723   809   887   923   1013   1091       {6}   651   729   815   893   929   1019   1097       {7}   701   781   865   943   981   1069   1147       {8}   635   712   799   877   913   1003   1081       {9}   631   709   795   873   909   999   1077       {10}    663   741   827   905   941   1031   1109                    13{1-7, 4, 1-10}, MS with positive APCI            13{x, 4, y}   x = {1}   {2}   {3}   {4}   {5}   {6}   {7}               Y = {1}   631   709   795   873   909   999   1077       {2}   649   727   813   891   927   1017   1095       {3}   661   739   825   903   939   1029   1107       {4}   699   777   863   941   977   1067   1145       {5}   659   737   823   901   937   1027   1105       {6}   665   743   829   907   943   1033   1111       {7}   715   793   879   957   993   1083   1161       {8}   649   727   813   891   927   989   1067       {9}   645   723   809   887   923   1013   1095       {10}    677   755   841   919   955   1045   1123                    13{1-7, 5, 1-10}, MS with positive APCI            13{x, 5, y}   x = {1}   {2}   {3}   {4}   {5}   {6}   {7}               Y {= {1}   645   723   809   887   923   1013   1091       {2}   663   741   827   905   941   1031   1109       {3}   675   753   839   917   953   1043   1121       {4}   713   791   877   955   991   1081   1159       {5}   673   751   837   915   951   1041   1119       {6}   679   757   843   921   957   1047   1125       {7}   729   807   893   971   1007   1097   1175       {8}   663   741   827   905   941   1031   1109       {9}   659   737   823   901   937   1027   1105       {10}    691   769   855   933   969   1059   1137                    13{1-7, 6, 1-10}, MS with positive APCI            13{x, 6, y}   x = {1}   {2}   {3}   {4}   {5}   {6}   {7}               Y = {1}   659   737   823   901   937   1027   1105       {2}   677   755   841   919   955   1045   1123       {3}   689   767   853   931   967   1057   1135       {4}   727   805   891   969   1005   1095   1173       {5}   687   765   851   929   965   1055   1133       {6}   693   771   857   935   971   1061   1111       {7}   743   821   907   985   1021   1111   1189       {8}   677   755   841   919   955   1045   1123       {9}   673   751   837   915   951   1041   1119       {10}    705   783   869   947   983   1073   1151                    13{1-7, 7, 1-10}, MS with positive APCI            13{x, 7, y}   x = {1}   {2}   {3}   {4}   {5}   {6}   {7}               Y = {1}   673   751   837   915   951   1041   1119       {2}   691   769   855   933   969   1059   1137       {3}   703   781   867   945   981   1071   1149       {4}   741   819   905   983   1019   1109   1187       {5}   701   779   865   943   979   1069   1147       {6}   707   784   871   949   985   1075   1153       {7}   757   835   921   999   1035   1125   1203       {8}   691   769   855   933   969   1059   1137       {9}   687   765   851   929   965   1055   1133       {10}   719   797   883   961   997   1087   1165                    13{1-7, 8, 1-10}, MS with positive APCI            13{x, 8, y}   x = {1}   {2}   {3}   {4}   {5}   {6}   {7}               Y = {1}   679   757   843   921   957   1047   1125       {2}   697   775   861   939   975   1065   1143       {3}   709   787   873   951   987   1077   1155       {4}   747   825   911   989   1025   1115   1193       {5}   707   785   871   949   985   1075   1153       {6}   713   791   877   955   991   1081   1159       {7}   763   841   927   1005    1041   1131   1209       {8}   697   775   861   939   975   1065   1143       {9}   693   771   857   935   971   1061   1139       {10}    725   803   889   967   1003   1093   1171                    
         [0215]    [0215]                                         TABLE 7                       LC-MS Data for Mappicine 5                                {5, 1, 2}   338   {2, 5, 1}   362   {7, 4, 6}   450       {3, 5, 9}   362   {5, 3, 7}   418   {3, 4, 9}   319       {4, 7, 3}   434   {2, 8, 5}   424   {4, 4, 3}   392       {1, 4, 7}   404   {3, 2, 1}   412   {1, 3, 3}   336       {2, 4, 8}   366   {1, 1, 3}   322   {3, 4, 2}   352       {5, 4, 4}   416   {4, 3, 2}   366   {3, 6, 10} a, b     425       {6, 4, 5}   416   {2, 4, 1}   348   {4, 6, 10} a, b     453       {5, 6, 1}   376   {4, 3, 6}   382   {7, 6, 10} a, b     507       {5, 2, 4}   494   {2, 3, 4}   402   {2, 1, 10} a     366       {3, 3, 5}   348   {6, 3, 8}   392   {5, 2, 10} a     473       {7, 3, 9}   416   {4, 4, 10} a     408   {7, 4, 10} a     462                                    
         [0216]    Although the present invention has been described in detail in connection with the above examples, it is to be understood that such detail is solely for that purpose and that variations can be made by those skilled in the art without departing from the spirit of the invention except as it may be limited by the following claims.