Abstract:
The invention relates to novel piperazine derivatives and their use as active ingredients in the preparation of pharmaceutical compositions. The invention also concerns related aspects including pharmaceutical compositions containing one or more of those compounds and their use as medicaments for the treatment or prevention of protozoal infections, such as especially malaria.

Description:
[0001]    The invention relates to novel compounds of the formula I. The invention also concerns related aspects including processes for the preparation of the compounds, pharmaceutical compositions containing one or more compounds of the formula I and especially their use as medicaments to treat or prevent malaria infections or to treat or prevent other protozoal diseases like sleeping sickness, Chagas disease, amebiasis, giardiasis, trichomoniasis, toxoplasmosis, and leishmaniasis. 
       BACKGROUND OF THE INVENTION 
       [0002]    Numerous serious diseases affecting humans as well as domestic and livestock animal are caused by protozoal organisms such as kinetoplastida, apicomplexa, anaerobic protozoa, microsporidia and plasmodium, for example. The clinically most relevant of these diseases is malaria. 
         [0003]    Malaria is one of the most serious and complex health problems affecting humanity in the 21 st  century. The disease affects about 300 million people worldwide, killing 1 to 1.5 million people every year. Malaria is an infectious disease caused by four species of the protozoan parasite plasmodium,  P. falciparum  being the most severe of the four. All attempts to develop vaccines against  P. falciparum  have failed so far. Therefore, therapies and preventive measures against malaria are confined to drugs. Various classes of antimalarial drugs exist. The most widely used are the quinoline antimalarials, e.g. chloroquine which has been an especially effective drug for both prophylaxis and therapy. However, resistance to many of the currently available antimalarial drugs is spreading rapidly, threatening people in areas where malaria is endemic. Reports of multi-drug resistant strains of malaria parasites render the search for new antimalarial agents especially urgent.  P. falciparum  enters the human body by way of bites of the female anophelino mosquito (it may also be transmitted by blood transfusion from asymptotic donors; almost all infected blood components including red cells, platelet concentrates, white cells, cryoprecipitates and fresh plasma can transmit malaria). The plasmodium parasite initially populates the liver, and during later stages of the infectious cycle reproduces in red blood cells. During this stage, the parasite degrades hemoglobin and uses the degradation products as nutrients for growth. 
         [0004]    The limitations of the current antiprotozoal chemotherapeutic arsenal underscore the need for new drugs in this therapeutic area. The present invention relates to the identification of novel low molecular weight, non-peptidic, non-quinoline compounds of formula I which are useful in the treatment and/or prevention of protozoal infections, especially in the treatment and/or prevention of malaria, in particular  plasmodium falciparum  malaria. 
         [0005]    WO 2007/046075 also discloses piperazine derivatives as antimalarial agents. The compound of WO 2007/046075 which comes closest to some of the presently claimed compounds is the compound of Example 54 which corresponds to reference Example 1 herein. However, the presently claimed compounds which come structurally closest to the compound of Example 54 of WO 2007/046075 exhibit an in vitro activity against erythrocytic stages of the  P. falciparum  strain NF54 in the presence of 50% serum which is significantly higher compared to the compound of Example 54 of WO 2007/046075 (see Table 1 below). 
     
    
     DETAILED DESCRIPTION OF THE INVENTION 
       [0006]    i) The present invention relates to novel compounds of the formula I: 
         [0000]    
       
                 
         
             
             
         
       
     
         [0000]    wherein
       X is CH or N;   R 1  represents —NO 2 , —N(CH 3 ) 2 , or —NCH 3 (CH 2 CH 2 OH); and   R 2  represents hydrogen, methyl, ethyl, n-propyl, isopropyl, tert-butyl, cyano, halogen, methoxy, ethoxy, n-propoxy, isopropoxy, trifluoromethyl, difluoromethoxy, methylsulfonyl, acetyl, or acetylamino; or   X is CH, R 1  is hydrogen, and R 2  is ethyl, isopropyl, tert-butyl, ethoxy, n-propoxy, isopropoxy, methylsulfonyl, acetylamino, or methoxycarbonyl; or   X is CH, R 1  is cyano, and R 2  is ethyl, isopropyl, tert-butyl, ethoxy, n-propoxy, isopropoxy, trifluoromethyl, difluoromethoxy, trifluoromethoxy, methylsulfonyl, or acetylamino; or   X is CH, R 1  is chloro, and R 2  is ethyl, isopropyl, tert-butyl, ethoxy, n-propoxy, isopropoxy, difluoromethoxy, methylsulfonyl, or acetylamino; or   X is CH, R 1  is methoxy or isopropoxy, and R 2  is trifluoromethyl; or   X is CH, R 1  is methylsulfonyl or ethylsulfonyl, and R 2  is trifluoromethyl, ethyl, isopropyl, tert-butyl, ethoxy, n-propoxy, isopropoxy, or difluoromethoxy, such as especially trifluoromethyl, tert-butyl, n-propoxy, or isopropoxy.       
 
         [0015]    ii) A further embodiment of the invention relates to compounds of the formula I according to embodiment i), wherein
       X is CH or N;   R 1  represents —NO 2 , —N(CH 3 ) 2 , or —NCH 3 (CH 2 CH 2 OH); and   R 2  represents hydrogen, methyl, ethyl, n-propyl, isopropyl, tert-butyl, cyano, halogen, methoxy, ethoxy, n-propoxy, isopropoxy, trifluoromethyl, difluoromethoxy, methylsulfonyl, acetyl, or acetylamino; or   X is CH, R 1  is hydrogen, and R 2  is ethyl, isopropyl, tert-butyl, ethoxy, n-propoxy, isopropoxy, methylsulfonyl, acetylamino, or methoxycarbonyl; or   X is CH, R 1  is cyano, and R 2  is ethyl, isopropyl, tert-butyl, ethoxy, n-propoxy, isopropoxy, trifluoromethyl, difluoromethoxy, trifluoromethoxy, methylsulfonyl, or acetylamino; or   X is CH, R 1  is chloro, and R 2  is ethyl, isopropyl, tert-butyl, ethoxy, n-propoxy, isopropoxy, difluoromethoxy, methylsulfonyl, or acetylamino; or   X is CH, R 1  is methoxy or isopropoxy, and R 2  is trifluoromethyl.       
 
         [0023]    iii) A further embodiment of the invention relates to compounds of the formula I according to embodiment i), wherein
       X is CH or N;   R 1  represents —NO 2 , —N(CH 3 ) 2 , or —NCH 3 (CH 2 CH 2 OH); and   R 2  represents ethyl, isopropyl, tert-butyl, methoxy, ethoxy, n-propoxy, isopropoxy, trifluoromethyl, difluoromethoxy, methylsulfonyl, or acetylamino; or   X is CH, R 1  is hydrogen, and R 2  is ethyl, isopropyl, tert-butyl, ethoxy, n-propoxy, isopropoxy, methylsulfonyl, acetylamino, or methoxycarbonyl; or   X is CH, R 1  is cyano, and R 2  is ethyl, isopropyl, tert-butyl, ethoxy, n-propoxy, isopropoxy, trifluoromethyl, difluoromethoxy, trifluoromethoxy, methylsulfonyl, or acetylamino; or   X is CH, R 1  is chloro, and R 2  is ethyl, isopropyl, tert-butyl, ethoxy, n-propoxy, isopropoxy, difluoromethoxy, methylsulfonyl, or acetylamino; or   X is CH, R 1  is methoxy or isopropoxy, and R 2  is trifluoromethyl.       
 
         [0031]    iv) A further embodiment of the invention relates to compounds of the formula I according to embodiment i), wherein 
         [0000]    R 1  represents —NO 2 , —N(CH 3 ) 2 , or —NCH 3 (CH 2 CH 2 OH); and
 
R 2  represents hydrogen, methyl, ethyl, n-propyl, isopropyl, tert-butyl, cyano, halogen, methoxy, ethoxy, n-propoxy, isopropoxy, trifluoromethyl, difluoromethoxy, methylsulfonyl, acetyl, or acetylamino.
 
         [0032]    v) A further embodiment of the invention relates to compounds of the formula I according to embodiment i), wherein 
         [0000]    R 1  represents —NO 2 , —N(CH 3 ) 2 , or —NCH 3 (CH 2 CH 2 OH); and
 
R 2  represents ethyl, isopropyl, tert-butyl, methoxy, ethoxy, n-propoxy, isopropoxy, trifluoromethyl, difluoromethoxy, methylsulfonyl, or acetylamino.
 
         [0033]    vi) A further embodiment of the invention relates to compounds of the formula I according to embodiment iv) or v), wherein X is CH. 
         [0034]    vii) A further embodiment of the invention relates to compounds of the formula I according to embodiment iv) or v), wherein X is N. 
         [0035]    viii) A further embodiment of the invention relates to compounds of the formula I according to any one of embodiments iv) to vii), wherein R 1  represents —NO 2 . 
         [0036]    ix) A further embodiment of the invention relates to compounds of the formula I according to any one of embodiments iv) to vii), wherein R 1  represents —N(CH 3 ) 2 . 
         [0037]    x) A further embodiment of the invention relates to compounds of the formula I according to any one of embodiments iv) to vii), wherein R 1  represents —NCH 3 (CH 2 CH 2 OH). 
         [0038]    xi) A further embodiment of the invention relates to compounds of the formula I according to any one of embodiments iv) to x), wherein R 2  is ethyl, isopropyl, tert-butyl, ethoxy, n-propoxy, or isopropoxy. 
         [0039]    xii) A further embodiment of the invention relates to compounds of the formula I according to embodiment xi), wherein R 2  is isopropoxy. 
         [0040]    xiii) A further embodiment of the invention relates to compounds of the formula I according to any one of embodiments iv) to x), wherein R 2  is trifluoromethyl, difluoromethoxy, methylsulfonyl, or acetylamino. 
         [0041]    xiv) A further embodiment of the invention relates to compounds of the formula I according to any one of embodiments iv) to x), wherein R 2  is methoxy. 
         [0042]    xv) A further embodiment of the invention relates to compounds of the formula I according to any one of embodiments iv) to x), wherein R 2  is hydrogen, methyl, n-propyl, cyano, halogen, or acetyl. 
         [0043]    The term “halogen” as used herein means fluorine, chlorine, bromine or iodine, such as especially fluorine or chlorine. 
         [0044]    Where the plural form is used for compounds, salts, pharmaceutical compositions, diseases and the like, this is intended to mean also a single compound, salt, or the like. 
         [0045]    Any reference hereinbefore or hereinafter to a compound of formula I is to be understood as referring also to salts, especially pharmaceutically acceptable salts, of a compound of formula I, as appropriate and expedient. 
         [0046]    The term “pharmaceutically acceptable salts” refers to non-toxic, inorganic or organic acid and/or base addition salts. Reference can be made to “Salt selection for basic drugs”,  Int. J. Pharm.  1986, 33, 201-17. 
         [0047]    The present invention also includes isotopically labelled, especially  2 H (deuterium) labelled compounds of formula I, which compounds are identical to the compounds of formula I except that one or more atoms have each been replaced by an atom having the same atomic number but an atomic mass different from the atomic mass usually found in nature. Isotopically labelled, especially  2 H (deuterium) labelled compounds of formula I and salts thereof are within the scope of the present invention. Substitution of hydrogen with the heavier isotope  2 H (deuterium) may lead to greater metabolic stability, resulting e.g. in increased in vivo half-life or reduced dosage requirements, or may lead to reduced inhibition of cytochrome P450 enzymes, resulting e.g. in an improved safety profile. In one embodiment of the invention, the compounds of formula I are not isotopically labelled, or they are labelled only with one or more deuterium atoms. In a sub-embodiment, the compounds of formula I are not isotopically labelled at all. Isotopically labelled compounds of formula I may be prepared in analogy to the methods described hereinafter, but using the appropriate isotopic variation of suitable reagents or starting materials. 
         [0048]    Examples of preferred compounds of formula I are selected from the group consisting of:
   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1-benzyl-2-[4-(4-dimethylamino-benzyl)-piperazin-1-yl]-2-oxo-ethyl}-3-(4-trifluoromethyl-phenyl)-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1-benzyl-2-[4-(4-dimethylamino-benzyl)-piperazin-1-yl]-2-oxo-ethyl}-3-(6-trifluoromethyl-pyridin-3-yl)-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1-benzyl-2-[4-(4-dimethylamino-benzyl)-piperazin-1-yl]-2-oxo-ethyl}-3-(6-methoxy-pyridin-3-yl)-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1-benzyl-2-[4-(4-dimethylamino-benzyl)-piperazin-1-yl]-2-oxo-ethyl}-3-(6-ethoxy-pyridin-3-yl)-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1-benzyl-2-[4-(4-dimethylamino-benzyl)-piperazin-1-yl]-2-oxo-ethyl}-3-(4-ethoxy-phenyl)-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1-benzyl-2-[4-(4-dimethylamino-benzyl)-piperazin-1-yl]-2-oxo-ethyl}-3-(4-methanesulfonyl-phenyl)-acrylamide,   (S)-3-(4-Acetylamino-phenyl)-N-[4-(4-acetyl-piperazin-1-yl)-benzyl]-N-{1-benzyl-2-[4-(4-dimethylamino-benzyl)piperazin-1-yl]-2-oxo-ethyl}-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1-benzyl-2-[4-(4-dimethylamino-benzyl)-piperazin-1-yl]-2-oxo-ethyl}-3-(4-propoxy-phenyl)-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1-benzyl-2-[4-(4-dimethylamino-benzyl)-piperazin-1-yl]-2-oxo-ethyl}-3-(4-isopropoxy-phenyl)-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1-benzyl-2-[4-(4-dimethylamino-benzyl)-piperazin-1-yl]-2-oxo-ethyl}-3-(4-ethyl-phenyl)-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1-benzyl-2-[4-(4-dimethylamino-benzyl)-piperazin-1-yl]-2-oxo-ethyl}-3-(4-difluoromethoxy-phenyl)-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1-benzyl-2-[4-(4-dimethylamino-benzyl)-piperazin-1-yl]-2-oxo-ethyl}-3-(4-tert-butyl-phenyl)-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1-benzyl-2-[4-(4-dimethylamino-benzyl)-piperazin-1-yl]-2-oxo-ethyl}-3-(4-isopropyl-phenyl)-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-[1-benzyl-2-(4-{4-[(2-hydroxy-ethyl)-methyl-amino]-benzyl}-piperazin-1-yl)-2-oxo-ethyl]-3-(4-trifluoromethyl-phenyl)-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-[1-benzyl-2-(4-{4-[(2-hydroxy-ethyl)-methyl-amino]-benzyl}-piperazin-1-yl)-2-oxo-ethyl]-3-(6-trifluoromethyl-pyridin-3-yl)-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-[1-benzyl-2-(4-{4-[(2-hydroxy-ethyl)-methyl-amino]-benzyl}-piperazin-1-yl)-2-oxo-ethyl]-3-(6-methoxy-pyridin-3-yl)-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-[1-benzyl-2-(4-{4-[(2-hydroxy-ethyl)-methyl-amino]-benzyl}-piperazin-1-yl)-2-oxo-ethyl]-3-(6-ethoxy-pyridin-3-yl)-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-[1-benzyl-2-(4-{4-[(2-hydroxy-ethyl)-methyl-amino]-benzyl}-piperazin-1-yl)-2-oxo-ethyl]-3-(4-ethoxy-phenyl)-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-[1-benzyl-2-(4-{4-[(2-hydroxy-ethyl)-methyl-amino]-benzyl}-piperazin-1-yl)-2-oxo-ethyl]-3-(4-methanesulfonyl-phenyl)-acrylamide,   (S)-3-(4-Acetylamino-phenyl)-N-[4-(4-acetyl-piperazin-1-yl)-benzyl]-N-[1-benzyl-2-(4-{4-[(2-hydroxy-ethyl)-methyl-amino]-benzyl}-piperazin-1-yl)-2-oxo-ethyl]-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-[1-benzyl-2-(4-{4-[(2-hydroxy-ethyl)-methyl-amino]-benzyl}-piperazin-1-yl)-2-oxo-ethyl]-3-(4-propoxy-phenyl)-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-[1-benzyl-2-(4-{4-[(2-hydroxy-ethyl)-methyl-amino]-benzyl}-piperazin-1-yl)-2-oxo-ethyl]-3-(4-isopropoxy-phenyl)-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-[1-benzyl-2-(4-{4-[(2-hydroxy-ethyl)-methyl-amino]-benzyl}-piperazin-1-yl)-2-oxo-ethyl]-3-(4-ethyl-phenyl)-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-[1-benzyl-2-(4-{4-[(2-hydroxy-ethyl)-methyl-amino]-benzyl}-piperazin-1-yl)-2-oxo-ethyl]-3-(4-difluoromethoxy-phenyl)-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-[1-benzyl-2-(4-{4-[(2-hydroxy-ethyl)-methyl-amino]-benzyl}-piperazin-1-yl)-2-oxo-ethyl]-3-(4-isopropyl-phenyl)-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-[1-benzyl-2-(4-{4-[(2-hydroxy-ethyl)-methyl-amino]-benzyl}-piperazin-1-yl)-2-oxo-ethyl]-3-(4-tert-butyl-phenyl)-acrylamide, and   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1-benzyl-2-[4-(4-nitro-benzyl)-piperazin-1-yl]-2-oxo-ethyl}-3-(4-trifluoromethyl-phenyl)-acrylamide.   
 
         [0076]    Further examples of preferred compounds of formula I are selected from the group consisting of:
   (S)—N-[4-(4-Acetyl-piperazin-1-yl)benzyl]-N-{1-(4-(4-(dimethylamino)benzyl)piperazin-1-yl)-1-oxo-3-phenylpropan-2-yl}cinnamamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)benzyl]-N-{1-[4-(4-(dimethylamino)benzyl)piperazin-1-yl]-1-oxo-3-phenylpropan-2-yl}-3-(p-tolyl)acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)benzyl]-N-{1-(4-(4-(dimethylamino)benzyl)piperazin-1-yl)-1-oxo-3-phenylpropan-2-yl}-3-(4-propylphenyl)acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)benzyl]-N-{1-(4-(4-(dimethylamino)benzyl)piperazin-1-yl)-1-oxo-3-phenylpropan-2-yl}-3-(4-methoxyphenyl)acrylamide,   (S)-3-(4-Acetyl-phenyl)-N-[4-(4-acetyl-piperazin-1-yl)benzyl]-N-{1-(4-(4-(dimethylamino)benzyl)piperazin-1-yl)-1-oxo-3-phenylpropan-2-yl}acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)benzyl]-3-(4-cyanophenyl)-N-{1-(4-(4-(dimethylamino)benzyl)piperazin-1-yl)-1-oxo-3-phenylpropan-2-yl}acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)benzyl]-N-{1-(4-(4-(dimethylamino)benzyl)piperazin-1-yl)-1-oxo-3-phenylpropan-2-yl}-3-(4-fluorophenyl)acrylamide, and   (S)—N-[4-(4-Acetyl-piperazin-1-yl)benzyl]-3-(4-chlorophenyl)-N-{1-(4-(4-(dimethylamino)benzyl)piperazin-1-yl)-1-oxo-3-phenylpropan-2-yl}acrylamide.   
 
         [0085]    Further examples of preferred compounds of formula I are selected from the group consisting of:
   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-[1-benzyl-2-(4-benzyl-piperazin-1-yl)-2-oxo-ethyl]-3-(4-ethyl-phenyl)-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-[1-benzyl-2-(4-benzyl-piperazin-1-yl)-2-oxo-ethyl]-3-(4-tert-butyl-phenyl)-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-[1-benzyl-2-(4-benzyl-piperazin-1-yl)-2-oxo-ethyl]-3-(4-ethoxy-phenyl)-acrylamide,   (S)-4-(2-{[4-(4-Acetyl-piperazin-1-yl)-benzyl]-[1-benzyl-2-(4-benzyl-piperazin-1-yl)-2-oxo-ethyl]carbamoyl}-vinyl)-benzoic acid methyl ester,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-[1-benzyl-2-(4-benzyl-piperazin-1-yl)-2-oxo-ethyl]-3-(4-methanesulfonyl-phenyl)-acrylamide,   (S)-3-(4-Acetylamino-phenyl)-N-[4-(4-acetyl-piperazin-1-yl)-benzyl]-N-[1-benzyl-2-(4-benzyl-piperazin-1-yl)-2-oxo-ethyl]-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-[1-benzyl-2-(4-benzyl-piperazin-1-yl)-2-oxo-ethyl]-3-(4-propoxy-phenyl)acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-[1-benzyl-2-(4-benzyl-piperazin-1-yl)-2-oxo-ethyl]-3-(4-isopropoxy-phenyl)-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-[1-benzyl-2-(4-benzyl-piperazin-1-yl)-2-oxo-ethyl]-3-(4-isopropyl-phenyl)-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1-benzyl-2-[4-(4-cyano-benzyl)-piperazin-1-yl]-2-oxo-ethyl}-3-(4-trifluoromethyl-phenyl)-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1-benzyl-2-[4-(4-cyano-benzyl)-piperazin-1-yl]-2-oxo-ethyl}-3-(4-ethoxy-phenyl)-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1-benzyl-2-[4-(4-cyano-benzyl)-piperazin-1-yl]-2-oxo-ethyl}-3-(4-methanesulfonyl-phenyl)acrylamide,   (S)-3-(4-Acetylamino-phenyl)-N-[4-(4-acetyl-piperazin-1-yl)-benzyl]-N-{1-benzyl-2-[4-(4-cyano-benzyl)-piperazin-1-yl]-2-oxo-ethyl}-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1-benzyl-2-[4-(4-cyano-benzyl)-piperazin-1-yl]-2-oxo-ethyl}-3-(4-propoxy-phenyl)-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1-benzyl-2-[4-(4-cyano-benzyl)-piperazin-1-yl]-2-oxo-ethyl}-3-(4-isopropoxy-phenyl)-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1-benzyl-2-[4-(4-cyano-benzyl)-piperazin-1-yl]-2-oxo-ethyl}-3-(4-ethyl-phenyl)-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1-benzyl-2-[4-(4-cyano-benzyl)-piperazin-1-yl]-2-oxo-ethyl}-3-(4-difluoromethoxy-phenyl)-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1-benzyl-2-[4-(4-cyano-benzyl)-piperazin-1-yl]-2-oxo-ethyl}-3-(4-isopropyl-phenyl)-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1-benzyl-2-[4-(4-cyano-benzyl)-piperazin-1-yl]-2-oxo-ethyl}-3-(4-trifluoromethoxy-phenyl)-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1-benzyl-2-[4-(4-cyano-benzyl)-piperazin-1-yl]-2-oxo-ethyl}-3-(4-tert-butyl-phenyl)-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1-benzyl-2-[4-(4-chloro-benzyl)-piperazin-1-yl]-2-oxo-ethyl}-3-(4-ethoxy-phenyl)-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1-benzyl-2-[4-(4-chloro-benzyl)-piperazin-1-yl]-2-oxo-ethyl}-3-(4-methanesulfonyl-phenyl)acrylamide,   (S)-3-(4-Acetylamino-phenyl)-N-[4-(4-acetyl-piperazin-1-yl)-benzyl]-N-{1-benzyl-2-[4-(4-chloro-benzyl)-piperazin-1-yl]-2-oxo-ethyl}-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1-benzyl-2-[4-(4-chloro-benzyl)-piperazin-1-yl]-2-oxo-ethyl}-3-(4-propoxy-phenyl)-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1-benzyl-2-[4-(4-chloro-benzyl)-piperazin-1-yl]-2-oxo-ethyl}-3-(4-isopropoxy-phenyl)-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1-benzyl-2-[4-(4-chloro-benzyl)-piperazin-1-yl]-2-oxo-ethyl}-3-(4-ethyl-phenyl)-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1-benzyl-2-[4-(4-chloro-benzyl)-piperazin-1-yl]-2-oxo-ethyl}-3-(4-difluoromethoxy-phenyl)-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1-benzyl-2-[4-(4-chloro-benzyl)-piperazin-1-yl]-2-oxo-ethyl}-3-(4-isopropyl-phenyl)-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1-benzyl-2-[4-(4-chloro-benzyl)-piperazin-1-yl]-2-oxo-ethyl}-3-(4-tert-butyl-phenyl)-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1-benzyl-2-[4-(4-methoxy-benzyl)-piperazin-1-yl]-2-oxo-ethyl}-3-(4-trifluoromethyl-phenyl)-acrylamide, and   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1-benzyl-2-[4-(4-isopropoxy-benzyl)-piperazin-1-yl]-2-oxo-ethyl}-3-(4-trifluoromethyl-phenyl)-acrylamide.   
 
         [0117]    Further examples of preferred compounds of formula I are selected from the group consisting of:
   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1-benzyl-2-[4-(4-(methylsulfonyl)-benzyl)-piperazin-1-yl]-2-oxo-ethyl}-3-(4-isopropoxyphenyl)-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1-benzyl-2-[4-(4-(methylsulfonyl)-benzyl)-piperazin-1-yl]-2-oxo-ethyl}-3-(4-tert-butyl-phenyl)-acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1-benzyl-2-[4-(4-(methylsulfonyl)-benzyl)-piperazin-1-yl]-2-oxo-ethyl}-3-(4-propoxyphenyl)-acrylamide,   (S)—N-(4-(4-acetylpiperazin-1-yl)benzyl)-N-(1-(4-(4-(methylsulfonyl)benzyl)piperazin-1-yl)-1-oxo-3-phenylpropan-2-yl)-3-(4-(trifluoromethyl)phenyl)acrylamide,   (S)—N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1-benzyl-2-[4-(4-(ethylsulfonyl)-benzyl)-piperazin-1-yl]-2-oxo-ethyl}-3-(4-propoxyphenyl)-acrylamide, and   (S)—N-(4-(4-acetylpiperazin-1-yl)benzyl)-N-(1-(4-(4-(ethylsulfonyl)benzyl)piperazin-1-yl)-1-oxo-3-phenylpropan-2-yl)-3-(4-(trifluoromethyl)phenyl)acrylamide.   
 
         [0124]    The compounds of formula I and their pharmaceutically acceptable salts can be used as medicaments, e.g. in the form of pharmaceutical compositions for enteral or parenteral administration, and are suitable for the treatment and/or prevention of the diseases mentioned herein, such as especially malaria. 
         [0125]    The production of the pharmaceutical compositions can be effected in a manner which will be familiar to any person skilled in the art (see for example Remington,  The Science and Practice of Pharmacy,  21st Edition (2005), Part 5, “Pharmaceutical Manufacturing” [published by Lippincott Williams &amp; Wilkins]) by bringing the described compounds of formula I or their pharmaceutically acceptable salts, optionally in combination with other therapeutically valuable substances, into a galenical administration form together with suitable, non-toxic, inert, pharmaceutically acceptable solid or liquid carrier materials and, if desired, usual pharmaceutical adjuvants. 
         [0126]    In one embodiment, the invention relates to a method for the treatment or prevention of the diseases mentioned herein, such as especially malaria, said method comprising administering to a subject a pharmaceutically active amount of a compound of formula I. 
         [0127]    The compounds of formula I or the above-mentioned pharmaceutical compositions may also be used in combination with one or more other therapeutically useful substances e.g. with other antimalarials like quinolines (e.g. quinine, chloroquine, amodiaquine, mefloquine, primaquine, and tafenoquine), peroxide antimalarials (e.g. artemisinin, artemether, and artesunate), pyrimethamine-sulfadoxine antimalarials (e.g. Fansidar®), hydroxynaphtoquinones (e.g. atovaquone), acroline-type antimalarials (e.g. pyronaridine), and other antiprotozoal agents like ethylstibamine, hydroxystilbamidine, pentamidine, stilbamidine, quinapyramine, puromycine, propamidine, nifurtimox, melarsoprol, nimorazole, nifuroxime, aminitrozole and the like. 
         [0128]    The present invention also relates to the use of a compound of formula I for the preparation of a pharmaceutical composition, optionally for use in combination with one or more other therapeutically useful substances such as those mentioned in the preceding paragraph, for the prevention and/or treatment of the diseases mentioned herein, such as especially malaria. 
         [0129]    The compounds of the formula I of the present invention may be prepared according to the procedures described herein, especially as described in the experimental part. 
         [0130]    In general, all chemical transformations can be performed according to well-known standard methodologies as described in the literature or as described in the procedures below. 
         [0131]    Preparation of compounds of formula I, except for compounds wherein R 1  is —NCH 3 (CH 2 CH 2 OH): 
         [0000]    
       
                 
         
             
             
         
       
       
                 
         
             
             
         
       
     
         [0132]    The Boc-Phe-OH 1 can be coupled with the benzylpiperazine derivative 2 via a peptidic coupling using activating agents such as TBTU (or PyBOP/HOBt) in the presence of a base such as DIPEA (or NEM) in DCM (or DMF) at RT to afford the intermediate 3. Alternatively, Cbz-Phe-OH can also be used in the initial peptidic coupling step to give 3. Boc-deprotection is usually achieved by reacting 3 with a solution of HCl 4N in dioxane using DCM as solvent, while Cbz-deprotection is achieved by hydrogenation with Pd/C catalyst in MeOH, to give the amine intermediate 4. Reductive amination between the free amine 4 and the aldehyde 5 at reflux in MeOH afforded the unstable imine (not depicted in the scheme), which is further reduced at RT with NaBH 4  to give the secondary amine intermediate 6. Alternatively, the reductive amination can be achieved in a solvent such as CH 3 CN in the presence of a reducing reagent such as NaBH(OAc) 3  to give the expected secondary amine intermediate 6. Compound 6 can then be coupled with a carboxylic acid 7 using a peptidic coupling reagent such as TBTU, PyBOP/HOBt or the Ghosez&#39;s reagent in a solvent such as DCM (or DMF) at RT in the presence of a base such as DIPEA (or NEM). Alternatively, the carboxylic acid 7 can be converted to the corresponding acid chloride (not depicted in the scheme) using oxalyl chloride in DCM to give the final compounds 8 of formula I. 
         [0133]    When R 1 =—NCH 3 (CH 2 CH 2 OH) the compounds of formula I are prepared according to Scheme 2. 
         [0000]    
       
                 
         
             
             
         
       
       
                 
         
             
             
         
       
     
         [0134]    The Boc-Phe-OH 1 is coupled with the benzylpiperazine derivative 9 via a peptidic coupling reaction using activating agents such as TBTU (or PyBOP/HOBt) in the presence of a base such as DIPEA (or NEM) in DCM (or DMF) at RT to afford the intermediate 10. Alternatively, Cbz-Phe-OH can also be used in the initial peptide coupling step to give 10. Boc-deprotection is usually achieved by reacting 10 with a solution of HCl 4N in dioxane using DCM as solvent, while Cbz-deprotection is achieved by hydrogenation with Pd/C catalyst in MeOH, to give the amine intermediate 11. Reductive amination between the free amine 11 and the aldehyde 5 in CH 3 CN at RT in the presence of a reducing agent such as NaBH(OAc) 3  affords the secondary amine intermediate 12. The free hydroxyl group of compound 12 is protected using for instance TBDMSCl as silylating agent to give 13, which is then coupled with a carboxylic acid 7 using a peptide coupling reagent such as TBTU (or PyBOP/HOBt) in a solvent such as DCM (or DMF) at RT in the presence of a base such as DIPEA. Alternatively, the carboxylic acid 7 can be activated by converting it to the corresponding acid chloride (not depicted in the scheme) using oxalyl chloride in DCM, to subsequently give compound 14. Further deprotection under mild acidic conditions such as aqueous 1 M HCl in MeOH or fluorinated reagents such as TBAF yield the final compounds 16 of formula I. 
         [0135]    The compounds of formula I can also be prepared according to the pathway depicted in Scheme 3. 
         [0000]    
       
                 
         
             
             
         
       
       
                 
         
             
             
         
       
     
         [0136]    Reductive amination between the amino acid H-Phe-OMe.HCl 17 and the aldehyde 5 in MeOH under reflux affords the corresponding imine, which is further reduced to the secondary amine 18 in the presence of a reducing reagent such as NaBH 4  at RT. 18 can also be obtained using the conditions described above for compounds 6 and 12. The ester 18 is then coupled with the acid chloride 19 derived from the carboxylic acid 7 using oxalyl chloride in DCM or Ghosez&#39;s reagent. Alternatively, 18 can be directly coupled with the carboxylic acid 7 via a peptide coupling reaction using TBTU (or PyBOP/HOBt) as coupling agents in a solvent such as DCM (or DMF) at RT in the presence of a base such as DIPEA (or NEM). Careful saponification of the ester 20 with aqueous LiOH 0.5 N in THF at 0° C. affords the acid 21. Final peptide coupling with the benzylpiperazine 2 gives the final compounds 8 of formula I. 
         [0137]    Benzylpiperazines 2 and 9 are commercially available and/or can be synthesized according to the following synthetic Scheme 4: 
         [0000]    
       
                 
         
             
             
         
       
     
         [0138]    Cinnamic acids 7 are commercially available or/and can be synthesized according to the following pathways: 
       Pathway A: Knoevenagel Reaction 
       [0139]    
       
                 
         
             
             
         
       
     
         [0140]    The cinnamic acids 7 are obtained by refluxing the aldehyde 25 with malonic acid in a mixture of piperidine/pyridine (WO 00/66566). 
       Pathway B: Horner-Emmons Reaction 
       [0141]    
       
                 
         
             
             
         
       
     
         [0142]    The cinnamic acids 7 are obtained in two steps by reacting the aldehydes 25 with triethyl phosphoacetate 26 in the presence of a base such as NaH in an aprotic solvent such as THF followed by saponification of the resulting ethyl ester with 4 N KOH in EtOH. 
         [0143]    The following examples illustrate the present invention. All temperatures are stated in degrees Celsius and pressures in mbar. Unless mentioned otherwise, the reactions take place at RT. The ratio of amounts of solvents to one another is always stated in parts by volume. Chemical names for final products and intermediates have been generated on the basis of the chemical structural formulae with the aid of ChemDrawPro Automatic Nomenclature program. 
       Analytic HPLC Conditions: 
       [0144]    (I) Agilent 1100 series with UV/Vis and MS detection (MS: Thermo Finnigan single quadrupole). Columns (4.6×50 mm, 5 μm): Waters X-Bridge C18 or Waters Atlantis T3. Basic conditions: eluents: A: MeCN, B: concentrated NH 3  in water (1.0 mL/L). Gradient 5 to 95% A over 1.5 min. Flow rate: 4.5 mL/min. 
         [0145]    Acidic conditions: eluents A: water+0.04% TFA, B: MeCN. Gradient 5 to 95% over 1.5 min. Flow rate 4.5 mL/min. 
       Preparative HPLC Conditions: 
       [0146]    Gilson with UV/Vis+MS or UV/Vis+ELSD detection. Basic conditions: eluents: A: MeCN, B: H 2 O+0.5% NH 3  (25% aqueous). 
         [0147]    (II) Waters X-Bridge column, 19×50 mm, 5 μm. Gradient: 20 to 90% A over 5 min. Flow rate: 40 mL/min. 
         [0148]    (III) Waters X-Bridge column, 30×75 mm, 10 μm. Gradient: 20 to 90% A over 6 min. Flow rate: 75 mL/min. 
         [0149]    The following abbreviations are used herein:
   AcOH acetic acid   Boc tert.-butyloxycarbonyl   Boc-Phe-OH Boc-L-phenylanaline   Cbz benzyloxycarbonyl   Cbz-Phe-OH Cbz-L-phenylanaline   DCM dichloromethane   DIPEA N,N-diisopropylethylamine   DMF N,N-dimethylformamide   Et ethyl   EtOH ethanol   EtOAc ethyl acetate   Et 2 O diethylether   ELSD evaporative light scattering detection   h hour(s)   HOBt hydroxybenzotriazole   H-Phe-OMe.HCl L-phenylalanine methylester hydrochloride   HPLC high performance liquid chromatography   LC-MS liquid chromatography-mass spectroscopy   Me methyl   MeOH methanol   min minute(s)   MS mass spectroscopy   NaBH(OAc) 3  sodium triacetoxyborohydride   NEM N-ethyl morpholine   PBS phosphate buffered saline   Pd/C palladium on carbon   PG protecting group   PyBOP benzotriazol-1-yl-oxy-tris-pyrrolidinophosphonium hexafluorophosphate   quant. quantitative   RT room temperature   Rt retention time of a substance in HPLC (in minutes)   TBAF tetra-n-butylammonium fluoride   TBDMSCl tert-butyldimethyl chlorosilane   TBTU O-benzotriazol-1-yl-N,N,N′,N′-tetramethyluronium tetrafluoroborate   TFA trifluoroacetic acid   THF tetrahydrofuran   UV ultra violet   V is visible   # number   
 
       Preparation of Compounds of formula I Via Pathway Depicted in Scheme 1 
     General Procedures and Examples 
     General Method A—Step 1 
       [0189]    
       
                 
         
             
             
         
       
     
         [0190]    To a stirred suspension of 1 mmol of Boc-Phe-OH or Cbz-Phe-OH in 0.6 mL of dry DCM (or DMF) under nitrogen are successively added 1 mmol of TBTU and 2 mmol of NEM. The resulting light yellow suspension is stirred at RT for 1 h before a solution of 1 mmol of benzylpiperazine in 0.25 mL of dry DCM (or DMF) is added. The obtained reaction mixture is further stirred at RT overnight. Upon completion the reaction is diluted with DCM and quenched with a saturated solution of NaHCO 3 . The aqueous phase is extracted with DCM (×3), the combined organic phases are successively washed with H 2 O and brine, dried over Na 2 SO 4 , filtered and concentrated under reduced pressure. The residue is purified by flash chromatography (SiO 2  60F) to afford the title compound. 
       Intermediates 1 
       [0191]      
         [0000]    
       
         
               
               
             
               
               
               
               
               
             
               
               
               
               
               
             
           
               
                   
                   
               
               
                   
                 LC-MS 
               
             
          
           
               
                 R 1 /PG 
                 Chemical name 
                 Yield 
                 Rt (min) 
                 [M + H] +   
               
               
                   
               
             
          
           
               
                 H/Boc 
                 (S)-[1-Benzyl-2-(4-benzyl-piperazin-1-yl)-2- 
                 Quant. 
                 0.98* 
                 424.22 
               
               
                   
                 oxo-ethyl]-carbamic acid tert-butyl ester 
               
               
                 CN/Boc 
                 (S)-{1-Benzyl-2-[4-(4-cyano-benzyl)-piperazin- 
                 94% 
                 0.92* 
                 449.12 
               
               
                   
                 1-yl]-2-oxo-ethyl}-carbamic acid tert-butyl ester 
               
               
                 N(CH 3 ) 2 / 
                 (S)-{1-Benzyl-2-[4-(4-dimethylamino-benzyl)- 
                 Quant. 
                 0.96* 
                 467.23 
               
               
                 Boc 
                 piperazin-1-yl]-2-oxo-ethyl}-carbamic acid tert- 
               
               
                   
                 butyl ester 
               
               
                 Cl/Boc 
                 (S)-{1-Benzyl-2-[4-(4-chloro-benzyl)-piperazin- 
                 Quant. 
                 1.01* 
                 457.79 
               
               
                   
                 1-yl]-2-oxo-ethyl}-carbamic acid tert-butyl ester 
               
               
                 S(O 2 )Me/ 
                 (S)-Benzyl (1-(4-(4- 
                 76% 
                 0.62** 
                 536.16 
               
               
                 Cbz 
                 (methylsulfonyl)benzyl)piperazin-1-yl)-1-oxo-3- 
               
               
                   
                 phenylpropan-2-yl)carbamate 
               
               
                 S(O 2 )Et/ 
                 (S)-Benzyl (1-(4-(4- 
                 47% 
                 0.64** 
                 550.17 
               
               
                 Cbz 
                 (ethylsulfonyl)benzyl)piperazin-1-yl)-1-oxo-3- 
               
               
                   
                 phenylpropan-2-yl)carbamate 
               
               
                   
               
               
                 *Analytic I, X-Bridge column, basic conditions. 
               
               
                 **Analytic, Waters Atlantis T3 column, acidic conditions 
               
             
          
         
       
     
       General Method B—Step 2 
       [0192]    
       
                 
         
             
             
         
       
     
       With PG=Boc: 
       [0193]    To a solution of 1 mmol of the Boc-protected amine in 9 mL of dry DCM at 0° C. are added dropwise 4.5 mL of HCl 4N in dioxane. The resulting reaction mixture is stirred at RT for 4 h under nitrogen atmosphere, cooled down to 0° C. and carefully neutralized to pH=7 with an aqueous solution of NaOH 1N. The aqueous phase is then extracted with DCM (×3). The combined organic phases are successively washed with H 2 O and brine, dried over Na 2 SO 4 , filtered and concentrated under reduced pressure to afford the free primary amine, which is used in the next step without further purification. 
       With PG=Cbz: 
       [0194]    A mixture of 2 mmol of the Cbz-protected amine, Pd—C 10% (100 mg) in dry EtOH (25 mL) is stirred at RT under hydrogen atmosphere for 3 h. The reaction mixture is filtered over Celite and concentrated under reduced pressure to afford the free primary amine, which is used in the next step without further purification. 
       Intermediates 2 
       [0195]      
         [0000]    
       
         
               
               
             
               
               
               
               
               
             
               
               
               
               
               
             
           
               
                   
                   
               
               
                   
                 LC-MS 
               
             
          
           
               
                 R 1   
                 Chemical name 
                 Yield 
                 Rt (min) 
                 [M + H] +   
               
               
                   
               
             
          
           
               
                 H 
                 (S)-2-Amino-1-(4-benzyl-piperazin-1-yl)-3- 
                 Quant. 
                 0.79* 
                 324.29 
               
               
                   
                 phenyl-propan-1-one 
               
               
                 CN 
                 (S)-4-[4-(2-Amino-3-phenyl-propionyl)- 
                 Quant. 
                 0.74* 
                 349.16 
               
               
                   
                 piperazin-1-ylmethyl]-benzonitrile 
               
               
                 N(CH 3 ) 2   
                 (S)-2-Amino-1-[4-(4-dimethylamino- 
                 Quant. 
                 0.78* 
                 367.19 
               
               
                   
                 benzyl)piperazin-1-yl]-3-phenyl-propan-1- 
               
               
                   
                 one 
               
               
                 Cl 
                 (S)-2-Amino-1-[4-(4-chloro-benzyl)- 
                 Quant. 
                 0.82* 
                 358.11 
               
               
                   
                 piperazin-1-yl]-3-phenyl-propan-1-one 
               
               
                 S(O 2 )Me 
                 (S)-2-Amino-1-(4-(4- 
                 89% 
                 0.37** 
                 401.79 
               
               
                   
                 (methylsulfonyl)benzyl)piperazin-1-yl)-3- 
               
               
                   
                 phenylpropan-1-one 
               
               
                 S(O 2 )Et 
                 (S)-2-Amino-1-(4-(4- 
                 88% 
                 0.4** 
                 416.07 
               
               
                   
                 (ethylsulfonyl)benzyl)piperazin-1-yl)-3- 
               
               
                   
                 phenylpropan-1-one 
               
               
                   
               
               
                 *Analytic I, X-Bridge column, basic conditions. 
               
               
                 **Analytic, Waters Atlantis T3 column, acidic conditions 
               
             
          
         
       
     
       General Methods C1 &amp; C2—Step 3 
       [0196]    
       
                 
         
             
             
         
       
     
       General Method C1: 
       [0197]    A solution of 1 mmol of amine and 1 mmol of aldehyde in 5 mL of dry MeOH is refluxed for 24 h under nitrogen. The resulting mixture is then cooled to RT prior to the addition of 1.5 mmol of NaBH 4  in portion. The obtained heterogeneous mixture is further stirred for 2 h at RT, quenched with a saturated aqueous solution of NaHCO 3  and extracted with EtOAc (×3). The combined organic phases are washed with brine, dried over Na 2 SO 4 , filtered and concentrated under reduced pressure. The residue is purified by flash chromatography (SiO 2  60F) to afford the title compound. 
       General Method C2: 
       [0198]    To a solution of 1 mmol of amine and 1 mmol of aldehyde in 5 mL of dry CH 3 CN are added portionwise 1.5 mmol of NaBH(OAc) 3 . The resulting heterogeneous mixture is further stirred for 4 h at RT, quenched with a saturated aqueous solution of NaHCO 3  and extracted with EtOAc (×3). The combined organic phases are washed with brine, dried over Na 2 SO 4 , filtered and concentrated under reduced pressure. The residue is purified by flash chromatography (SiO 2  60F) to afford the title compound. 
       Intermediates 3 
       [0199]      
         [0000]    
       
         
               
               
             
               
               
               
               
               
             
               
               
               
               
               
             
           
               
                   
                   
               
               
                   
                 LC-MS 
               
             
          
           
               
                 R 1   
                 Chemical name 
                 Yield 
                 Rt (min) 
                 [M + H] +   
               
               
                   
               
             
          
           
               
                 H 
                 (S)-2-[4-(4-Acetyl-piperazin-1-yl)- 
                 77% 
                 0.84* 
                 540.30 
               
               
                   
                 benzylamino]-1-(4-benzyl-piperazin-1- 
               
               
                   
                 yl)-3-phenyl-propan-1-one 
               
               
                 CN 
                 (S)-4-(4-{2-[4-(4-Acetyl-piperazin-1- 
                 81% 
                 0.79* 
                 565.16 
               
               
                   
                 yl)-benzylamino]-3-phenyl-propionyl}- 
               
               
                   
                 piperazin-1-ylmethyl)-benzonitrile 
               
               
                 N(CH 3 ) 2   
                 (S)-2-[4-(4-Acetyl-piperazin-1-yl)- 
                 64% 
                 0.83* 
                 583.20 
               
               
                   
                 benzylamino]-1-[4-(4-dimethylamino- 
               
               
                   
                 benzyl)-piperazin-1-yl]-3-phenyl- 
               
               
                   
                 propan-1-one 
               
               
                 Cl 
                 (S)-2-[4-(4-Acetyl-piperazin-1-yl)- 
                 76% 
                 0.88* 
                 574.10 
               
               
                   
                 benzylamino]-1-[4-(4-chloro-benzyl)- 
               
               
                   
                 piperazin-1-yl]-3-phenyl-propan-1-one 
               
               
                 S(O 2 )Me 
                 (S)-2-[4-(4-Acetyl-piperazin-1-yl)- 
                 52% 
                 0.43** 
                 618.22 
               
               
                   
                 benzylamino]-1-[4-(4-methylsulfonyl- 
               
               
                   
                 benzyl)-piperazin-1-yl]-3-phenyl- 
               
               
                   
                 propan-1-one 
               
               
                 S(O 2 )Et 
                 (S)-2-[4-(4-Acetyl-piperazin-1-yl)- 
                 80% 
                 0.45** 
                 632.2 
               
               
                   
                 benzylamino]-1-[4-(4-ethylsulfonyl- 
               
               
                   
                 benzyl)-piperazin-1-yl]-3-phenyl- 
               
               
                   
                 propan-1-one 
               
               
                   
               
               
                 *Analytic I, X-Bridge column, basic conditions. 
               
               
                 **Analytic, Waters Atlantis T3 column, acidic conditions 
               
             
          
         
       
     
       General Methods D1 and D2—Step 4 
       [0200]    
       
                 
         
             
             
         
       
     
       General Method D1: 
       [0201]    To a solution of 1 mmol of cinnamic acid in 5 mL of dry DCM under nitrogen are added 1.4 mmol of 1-chloro-N,N-2-trimethylpropenylamine (Ghosez&#39;s reagent). The resulting mixture is stirred at RT for 1 h before a solution of 1 mmol of amine and 3 mmol of DIPEA in 4 mL of dry DCM is added. The reaction mixture is further stirred at RT overnight. Upon completion a saturated aqueous solution of NaHCO 3  is added and the mixture extracted with DCM (×3). The combined organic phases are washed with brine, dried over Na 2 SO 4 , filtered and concentrated under reduced pressure. The residue is purified either by flash chromatography (SiO 2  60F) or by preparative HPLC to afford the final compound. 
       General Method D2: 
       [0202]    To a solution (or suspension) of 1.05 mmol of cinnamic acid in 3.5 mL of dry DCM under nitrogen at 0° C. are added 1.1 mmol of oxalyl chloride and 3 drops of DMF. The resulting mixture is stirred at RT for 1 h, cooled down to 0° C. before a solution of 1 mmol of amine and 2 mmol of DIPEA in 3 mL of dry DCM is added. The reaction mixture is further stirred at RT overnight. Upon completion a saturated aqueous solution of NaHCO 3  is added and the mixture extracted with DCM (×3). The combined organic phases are washed with brine, dried over Na 2 SO 4 , filtered and concentrated under reduced pressure. The residue is purified either by flash chromatography (SiO 2  60F) or by preparative HPLC to afford the final compound. 
         [0000]    
       
         
               
               
               
             
               
               
               
               
             
               
               
               
               
             
           
               
                   
               
               
                 Compound of 
                   
                 LC-MS 
               
             
          
           
               
                 Example # 
                 Chemical name 
                 Rt (min) 
                 [M + H] +   
               
               
                   
               
             
          
           
               
                   1** 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-[1- 
                 1.00* 
                 738.37 
               
               
                   
                 benzyl-2-(4-benzyl-piperazin-1-yl)-2-oxo-ethyl]-3- 
               
               
                   
                 (4-trifluoromethyl-phenyl)-acrylamide 
               
               
                  2 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-[1- 
                 1.02* 
                 698.45 
               
               
                   
                 benzyl-2-(4-benzyl-piperazin-1-yl)-2-oxo-ethyl]-3- 
               
               
                   
                 (4-ethyl-phenyl)-acrylamide 
               
               
                  3 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-[1- 
                 1.06* 
                 726.46 
               
               
                   
                 benzyl-2-(4-benzyl-piperazin-1-yl)-2-oxo-ethyl]-3- 
               
               
                   
                 (4-tert-butyl-phenyl)-acrylamide 
               
               
                  4 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-[1- 
                 0.98* 
                 714.26 
               
               
                   
                 benzyl-2-(4-benzyl-piperazin-1-yl)-2-oxo-ethyl]-3- 
               
               
                   
                 (4-ethoxy-phenyl)-acrylamide 
               
               
                  5 
                 (S)-4-(2-{[4-(4-Acetyl-piperazin-1-yl)-benzyl]-[1- 
                 0.94* 
                 728.39 
               
               
                   
                 benzyl-2-(4-benzyl-piperazin-1-yl)-2-oxo-ethyl]- 
               
               
                   
                 carbamoyl}-vinyl)-benzoic acid methyl ester 
               
               
                  6 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-[1- 
                 0.88* 
                 748.36 
               
               
                   
                 benzyl-2-(4-benzyl-piperazin-1-yl)-2-oxo-ethyl]-3- 
               
               
                   
                 (4-methanesulfonyl-phenyl)-acrylamide 
               
               
                  7 
                 (S)-3-(4-Acetylamino-phenyl)-N-[4-(4-acetyl- 
                 0.86* 
                 727.37 
               
               
                   
                 piperazin-1-yl)benzyl]-N-[1-benzyl-2-(4-benzyl- 
               
               
                   
                 piperazin-1-yl)-2-oxo-ethyl]-acrylamide 
               
               
                  8 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-[1- 
                 1.02* 
                 728.40 
               
               
                   
                 benzyl-2-(4-benzyl-piperazin-1-yl)-2-oxo-ethyl]-3- 
               
               
                   
                 (4-propoxy-phenyl)-acrylamide 
               
               
                  9 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-[1- 
                 1.00* 
                 728.43 
               
               
                   
                 benzyl-2-(4-benzyl-piperazin-1-yl)-2-oxo-ethyl]-3- 
               
               
                   
                 (4-isopropoxy-phenyl)-acrylamide 
               
               
                 10 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-[1- 
                 1.05* 
                 712.28 
               
               
                   
                 benzyl-2-(4-benzyl-piperazin-1-yl)-2-oxo-ethyl]-3- 
               
               
                   
                 (4-isopropyl-phenyl)-acrylamide 
               
               
                 11 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1- 
                 0.99* 
                 781.18 
               
               
                   
                 benzyl-2-[4-(4-dimethylamino-benzyl)-piperazin-1- 
               
               
                   
                 yl]-2-oxo-ethyl}-3-(4-trifluoromethyl-phenyl)- 
               
               
                   
                 acrylamide 
               
               
                 12 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1- 
                 0.94* 
                 782.19 
               
               
                   
                 benzyl-2-[4-(4-dimethylamino-benzyl)-piperazin-1- 
               
               
                   
                 yl]-2-oxo-ethyl}-3-(6-trifluoromethyl-pyridin-3-yl)- 
               
               
                   
                 acrylamide 
               
               
                 13 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1- 
                 0.91* 
                 744.20 
               
               
                   
                 benzyl-2-[4-(4-dimethylamino-benzyl)-piperazin-1- 
               
               
                   
                 yl]-2-oxo-ethyl}-3-(6-methoxy-pyridin-3-yl)- 
               
               
                   
                 acrylamide 
               
               
                 14 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1- 
                 0.94* 
                 758.22 
               
               
                   
                 benzyl-2-[4-(4-dimethylamino-benzyl)-piperazin-1- 
               
               
                   
                 yl]-2-oxo-ethyl}-3-(6-ethoxy-pyridin-3-yl)-acrylamide 
               
               
                 15 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1- 
                 0.97* 
                 757.17 
               
               
                   
                 benzyl-2-[4-(4-dimethylamino-benzyl)-piperazin-1- 
               
               
                   
                 yl]-2-oxo-ethyl}-3-(4-ethoxy-phenyl)-acrylamide 
               
               
                 16 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1- 
                 0.87* 
                 791.15 
               
               
                   
                 benzyl-2-[4-(4-dimethylamino-benzyl)-piperazin-1- 
               
               
                   
                 yl]-2-oxo-ethyl}-3-(4-methanesulfonyl-phenyl)- 
               
               
                   
                 acrylamide 
               
               
                 17 
                 (S)-3-(4-Acetylamino-phenyl)-N-[4-(4-acetyl- 
                 0.85* 
                 770.22 
               
               
                   
                 perazin-1-yl)-benzyl]-N-{1-benzyl-2-[4-(4- 
               
               
                   
                 dimethylamino-benzyl)-piperazin-1-yl]-2-oxo-ethyl}- 
               
               
                   
                 acrylamide 
               
               
                 18 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1- 
                 1.00* 
                 771.27 
               
               
                   
                 benzyl-2-[4-(4-dimethylamino-benzyl)-piperazin-1- 
               
               
                   
                 yl]-2-oxo-ethyl}-3-(4-propoxy-phenyl)-acrylamide 
               
               
                 19 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1- 
                 0.99* 
                 771.25 
               
               
                   
                 benzyl-2-[4-(4-dimethylamino-benzyl)-piperazin-1- 
               
               
                   
                 yl]-2-oxo-ethyl}-3-(4-isopropoxy-phenyl)-acrylamide 
               
               
                 20 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1- 
                 1.00* 
                 741.28 
               
               
                   
                 benzyl-2-[4-(4-dimethylamino-benzyl)-piperazin-1- 
               
               
                   
                 yl]-2-oxo-ethyl}-3-(4-ethyl-phenyl)-acrylamide 
               
               
                 21 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1- 
                 0.95* 
                 779.15 
               
               
                   
                 benzyl-2-[4-(4-dimethylamino-benzyl)-piperazin-1- 
               
               
                   
                 yl]-2-oxo-ethyl}-3-(4-difluoromethoxy-phenyl)- 
               
               
                   
                 acrylamide 
               
               
                 22 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1- 
                 1.05* 
                 769.26 
               
               
                   
                 benzyl-2-[4-(4-dimethylamino-benzyl)-piperazin-1- 
               
               
                   
                 yl]-2-oxo-ethyl}-3-(4-tert-butyl-phenyl)-acrylamide 
               
               
                 23 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1- 
                 1.03* 
                 755.28 
               
               
                   
                 benzyl-2-[4-(4-dimethylamino-benzyl)-piperazin-1- 
               
               
                   
                 yl]-2-oxo-ethyl}-3-(4-isopropyl-phenyl)-acrylamide 
               
               
                 24 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1- 
                 0.96* 
                 763.11 
               
               
                   
                 benzyl-2-[4-(4-cyano-benzyl)-piperazin-1-yl]-2-oxo- 
               
               
                   
                 ethyl}-3-(4-trifluoromethyl-phenyl)-acrylamide 
               
               
                 25 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1- 
                 0.93* 
                 739.22 
               
               
                   
                 benzyl-2-[4-(4-cyano-benzyl)-piperazin-1-yl]-2-oxo- 
               
               
                   
                 ethyl}-3-(4-ethoxy-phenyl)-acrylamide 
               
               
                 26 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1- 
                 0.84* 
                 773.00 
               
               
                   
                 benzyl-2-[4-(4-cyano-benzyl)-piperazin-1-yl]-2-oxo- 
               
               
                   
                 ethyl}-3-(4-methanesulfonyl-phenyl)-acrylamide 
               
               
                 27 
                 (S)-3-(4-Acetylamino-phenyl)-N-[4-(4-acetyl- 
                 0.82* 
                 752.02 
               
               
                   
                 piperazin-1-yl)-benzyl]-N-{1-benzyl-2-[4-(4-cyano- 
               
               
                   
                 benzyl)piperazin-1-yl]-2-oxo-ethyl}-acrylamide 
               
               
                 28 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1- 
                 0.97* 
                 753.15 
               
               
                   
                 benzyl-2-[4-(4-cyano-benzyl)-piperazin-1-yl]-2-oxo- 
               
               
                   
                 ethyl}-3-(4-propoxy-phenyl)-acrylamide 
               
               
                 29 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1- 
                 0.96* 
                 753.18 
               
               
                   
                 benzyl-2-[4-(4-cyano-benzyl)-piperazin-1-yl]-2-oxo- 
               
               
                   
                 ethyl}-3-(4-isopropoxy-phenyl)-acrylamide 
               
               
                 30 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1- 
                 0.97* 
                 723.19 
               
               
                   
                 benzyl-2-[4-(4-cyano-benzyl)-piperazin-1-yl]-2-oxo- 
               
               
                   
                 ethyl}-3-(4-ethyl-phenyl)-acrylamide 
               
               
                 31 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1- 
                 0.92* 
                 761.06 
               
               
                   
                 benzyl-2-[4-(4-cyano-benzyl)-piperazin-1-yl]-2-oxo- 
               
               
                   
                 ethyl}-3-(4-difluoromethoxy-phenyl)-acrylamide 
               
               
                 32 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1- 
                 0.99* 
                 737.23 
               
               
                   
                 benzyl-2-[4-(4-cyano-benzyl)-piperazin-1-yl]-2-oxo- 
               
               
                   
                 ethyl}-3-(4-isopropyl-phenyl)-acrylamide 
               
               
                 33 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1- 
                 0.97* 
                 778.97 
               
               
                   
                 benzyl-2-[4-(4-cyano-benzyl)-piperazin-1-yl]-2-oxo- 
               
               
                   
                 ethyl}-3-(4-trifluoromethoxy-phenyl)-acrylamide 
               
               
                 34 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1- 
                 1.02* 
                 751.21 
               
               
                   
                 benzyl-2-[4-(4-cyano-benzyl)-piperazin-1-yl]-2-oxo- 
               
               
                   
                 ethyl}-3-(4-tert-butyl-phenyl)-acrylamide 
               
               
                 35 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1- 
                 1.00* 
                 748.13 
               
               
                   
                 benzyl-2-[4-(4-chloro-benzyl)-piperazin-1-yl]-2-oxo- 
               
               
                   
                 ethyl}-3-(4-ethoxy-phenyl)-acrylamide 
               
               
                 36 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1- 
                 0.90* 
                 782.11 
               
               
                   
                 benzyl-2-[4-(4-chloro-benzyl)-piperazin-1-yl]-2-oxo- 
               
               
                   
                 ethyl}-3-(4-methanesulfonyl-phenyl)-acrylamide 
               
               
                 37 
                 (S)-3-(4-Acetylamino-phenyl)-N-[4-(4-acetyl- 
                 0.88* 
                 761.18 
               
               
                   
                 piperazin-1-yl)-benzyl]-N-{1-benzyl-2-[4-(4-chloro- 
               
               
                   
                 benzyl)-piperazin-1-yl]-2-oxo-ethyl}-acrylamide 
               
               
                 38 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1- 
                 1.03* 
                 762.18 
               
               
                   
                 benzyl-2-[4-(4-chloro-benzyl)-piperazin-1-yl]-2-oxo- 
               
               
                   
                 ethyl}-3-(4-propoxy-phenyl)-acrylamide 
               
               
                 39 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1- 
                 1.02* 
                 762.13 
               
               
                   
                 benzyl-2-[4-(4-chloro-benzyl)-piperazin-1-yl]-2-oxo- 
               
               
                   
                 ethyl}-3-(4-isopropoxy-phenyl)-acrylamide 
               
               
                 40 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1- 
                 1.04* 
                 732.25 
               
               
                   
                 benzyl-2-[4-(4-chloro-benzyl)-piperazin-1-yl]-2-oxo- 
               
               
                   
                 ethyl}-3-(4-ethyl-phenyl)-acrylamide 
               
               
                 41 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1- 
                 0.98* 
                 770.15 
               
               
                   
                 benzyl-2-[4-(4-chloro-benzyl)-piperazin-1-yl]-2-oxo- 
               
               
                   
                 ethyl}-3-(4-difluoromethoxy-phenyl)-acrylamide 
               
               
                 42 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1- 
                 1.06* 
                 745.96 
               
               
                   
                 benzyl-2-[4-(4-chloro-benzyl)-piperazin-1-yl]-2-oxo- 
               
               
                   
                 ethyl}-3-(4-isopropyl-phenyl)-acrylamide 
               
               
                 43 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1- 
                 1.08* 
                 760.13 
               
               
                   
                 benzyl-2-[4-(4-chloro-benzyl)-piperazin-1-yl]-2-oxo- 
               
               
                   
                 ethyl}-3-(4-tert-butyl-phenyl)-acrylamide 
               
               
                 44 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1- 
                 0.73*** 
                 806.37 
               
               
                   
                 benzyl-2-[4-(4-(methylsulfonyl)-benzyl)-piperazin-1- 
               
               
                   
                 yl]-2-oxo-ethyl}-3-(4-isopropoxyphenyl)-acrylamide 
               
               
                 45 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1- 
                 0.78*** 
                 804.37 
               
               
                   
                 benzyl-2-[4-(4-(methylsulfonyl)-benzyl)-piperazin-1- 
               
               
                   
                 yl]-2-oxo-ethyl}-3-(4-tert-butyl-phenyl)-acrylamide 
               
               
                 46 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1- 
                 0.76*** 
                 806.37 
               
               
                   
                 benzyl-2-[4-(4-(methylsulfonyl)-benzyl)-piperazin-1- 
               
               
                   
                 yl]-2-oxo-ethyl}-3-(4-propoxyphenyl)-acrylamide 
               
               
                 47 
                 (S)-N-(4-(4-acetylpiperazin-1-yl)benzyl)-N-(1-(4-(4- 
                 0.75*** 
                 816.23 
               
               
                   
                 (methylsulfonyl)benzyl)piperazin-1-yl)-1-oxo-3- 
               
               
                   
                 phenylpropan-2-yl)-3-(4- 
               
               
                   
                 (trifluoromethyl)phenyl)acrylamide 
               
               
                 48 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1- 
                 0.77*** 
                 820.43 
               
               
                   
                 benzyl-2-[4-(4-(ethylsulfonyl)-benzyl)-piperazin-1- 
               
               
                   
                 yl]-2-oxo-ethyl}-3-(4-propoxyphenyl)-acrylamide 
               
               
                 49 
                 (S)-N-(4-(4-acetylpiperazin-1-yl)benzyl)-N-(1-(4-(4- 
                 0.76*** 
                 829.85 
               
               
                   
                 (ethylsulfonyl)benzyl)piperazin-1-yl)-1-oxo-3- 
               
               
                   
                 phenylpropan-2-yl)-3-(4- 
               
               
                   
                 (trifluoromethyl)phenyl)acrylamide 
               
               
                   
               
               
                 *Analytic I, X-Bridge column, basic conditions. 
               
               
                 **Reference Example 
               
               
                 ***Analytic, Waters Atlantis T3 column, acidic conditions 
               
             
          
         
       
     
       Preparation of Compounds of formula I Via Pathway Depicted in Scheme 2 
     General Procedures and Examples 
     Step 1: (S)-[1-Benzyl-2-(4-{4-[(2-hydroxy-ethyl)-methyl-amino]-benzyl}-piperazin-1-yl)-2-oxo-ethyl]-carbamic acid tert-butyl ester 
       [0203]    
       
                 
         
             
             
         
       
     
         [0204]    According to general Method A, 60 mmol of Boc-Phe-OH are used to provide the title compound in 50% yield. Rt=0.87; [M+H] + =497.42 (Analytic I, X-Bridge column, basic conditions). 
       Step 2: (S)-2-Amino-1-(4-{4-[(2-hydroxy-ethyl)-methyl-amino]-benzyl}-piperazin-1-yl)-3-phenyl-propan-1-one 
       [0205]    
       
                 
         
             
             
         
       
     
         [0206]    According to general Method B, 12 mmol of Boc-protected amine 10 are used to provide the title compound in quantitative yield. Rt=0.69; [M+H] + =397.18 (Analytic I, X-Bridge column, basic conditions). 
       Step 3: (S)-2-[4-(4-Acetyl-piperazin-1-yl)-benzylamino]-1-(4-{4-[(2-hydroxy-ethyl)-methyl-amino]-benzyl}-piperazin-1-yl)-3-phenyl-propan-1-one 
       [0207]    
       
                 
         
             
             
         
       
     
         [0208]    According to general Method C2, 11.4 mmol of free amine 11 are used to provide the title compound in quantitative yield. Rt=0.74; [M+H] + =613.24 (Analytic I, X-Bridge column, basic conditions). 
       General Method E—step 4: (S)-2-[4-(4-Acetyl-piperazin-1-yl)-benzylamino]-1-[4-(4-{[2-(tert-butyl-dimethyl-silanyloxy)-ethyl]-methyl-amino}-benzyl)-piperazin-1-yl]-3-phenyl-propan-1-one 
       [0209]    
       
                 
         
             
             
         
       
     
         [0210]    3 mmol of TBDMSCl are added portionwise to a solution of 1 mmol of hydroxyl 12 and 3 mmol of imidazole in 5 mL of dry DMF at RT. The yellow solution is stirred at RT for 16 h, quenched with H 2 O and extracted with EtOAc (×3). The combined organic phases are washed with brine, dried over Na 2 SO 4 , filtered and concentrated under reduced pressure. The residue is purified by flash chromatography (SiO 2  60F; DCM/MeOH 92:8 to 95:5) to afford the title compound as yellow foam in 81% yield. Rt=1.12 min; [M+H] + =727.31. (Analytic I, X-Bridge column, basic conditions). 
       Step 5 
       [0211]    
       
                 
         
             
             
         
       
     
         [0212]    The compounds of formula 28 are obtained according to general Methods D1 or D2. 
         [0000]    
       
         
               
               
             
               
               
               
               
             
               
               
               
               
             
           
               
                   
                   
               
               
                   
                 LC-MS* 
               
             
          
           
               
                 Intermediate # 
                 Chemical name 
                 Rt (min) 
                 [M + H] +   
               
               
                   
               
             
          
           
               
                 4 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1- 
                 1.21 
                 925.21 
               
               
                   
                 benzyl-2-[4-(4-{[2-(tert-butyl-dimethyl-silanyloxy)- 
               
               
                   
                 ethyl]-methyl-amino}-benzyl)-piperazin-1-yl]-2-oxo- 
               
               
                   
                 ethyl}-3-(4-trifluoromethyl-phenyl)-acrylamide 
               
               
                 5 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1- 
                 1.15 
                 926.27 
               
               
                   
                 benzyl-2-[4-(4-{[2-(tert-butyl-dimethyl-silanyloxy)- 
               
               
                   
                 ethyl]-methyl-amino}-benzyl)-piperazin-1-yl]-2-oxo- 
               
               
                   
                 ethyl}-3-(6-trifluoromethyl-pyridin-3-yl)-acrylamide 
               
               
                 6 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1- 
                 1.14 
                 888.29 
               
               
                   
                 benzyl-2-[4-(4-{[2-(tert-butyl-dimethyl-silanyloxy)- 
               
               
                   
                 ethyl]-methyl-amino}-benzyl)-piperazin-1-yl]-2-oxo- 
               
               
                   
                 ethyl}-3-(6-methoxy-pyridin-3-yl)-acrylamide 
               
               
                 7 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1- 
                 1.19 
                 902.33 
               
               
                   
                 benzyl-2-[4-(4-{[2-(tert-butyl-dimethyl-silanyloxy)- 
               
               
                   
                 ethyl]-methyl-amino}-benzyl)-piperazin-1-yl]-2-oxo- 
               
               
                   
                 ethyl}-3-(6-ethoxy-pyridin-3-yl)-acrylamide 
               
               
                 8 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1- 
                 1.20 
                 901.29 
               
               
                   
                 benzyl-2-[4-(4-{[2-(tert-butyl-dimethyl-silanyloxy)- 
               
               
                   
                 ethyl]-methyl-amino}-benzyl)-piperazin-1-yl]-2-oxo- 
               
               
                   
                 ethyl}-3-(4-ethoxy-phenyl)-acrylamide 
               
               
                 9 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1- 
                 1.09 
                 935.21 
               
               
                   
                 benzyl-2-[4-(4-{[2-(tert-butyl-dimethyl-silanyloxy)- 
               
               
                   
                 ethyl]-methyl-amino}-benzyl)-piperazin-1-yl]-2-oxo- 
               
               
                   
                 ethyl}-3-(4-methanesulfonyl-phenyl)-acrylamide 
               
               
                 10 
                 (S)-3-(4-Acetylamino-phenyl)-N-[4-(4-acetyl- 
                 1.08 
                 914.29 
               
               
                   
                 piperazin-1-yl)-benzyl]-N-{1-benzyl-2-[4-(4-{[2-(tert- 
               
               
                   
                 butyl-dimethyl-silanyloxy)-ethyl]-methyl-amino}- 
               
               
                   
                 benzyl)-piperazin-1-yl]-2-oxo-ethyl}-acrylamide 
               
               
                 11 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1- 
                 1.24 
                 915.28 
               
               
                   
                 benzyl-2-[4-(4-{[2-(tert-butyl-dimethyl-silanyloxy)- 
               
               
                   
                 ethyl]-methyl-amino}-benzyl)-piperazin-1-yl]-2-oxo- 
               
               
                   
                 ethyl}-3-(4-propoxy-phenyl)-acrylamide 
               
               
                 12 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1- 
                 1.22 
                 915.32 
               
               
                   
                 benzyl-2-[4-(4-{[2-(tert-butyl-dimethyl-silanyloxy)- 
               
               
                   
                 ethyl]-methyl-amino}-benzyl)-piperazin-1-yl]-2-oxo- 
               
               
                   
                 ethyl}-3-(4-isopropoxy-phenyl)-acrylamide 
               
               
                 13 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1- 
                 1.25 
                 885.35 
               
               
                   
                 benzyl-2-[4-(4-{[2-(tert-butyl-dimethyl-silanyloxy)- 
               
               
                   
                 ethyl]-methyl-amino}-benzyl)-piperazin-1-yl]-2-oxo- 
               
               
                   
                 ethyl}-3-(4-ethyl-phenyl)-acrylamide 
               
               
                 14 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1- 
                 1.15 
                 923.21 
               
               
                   
                 benzyl-2-[4-(4-{[2-(tert-butyl-dimethyl-silanyloxy)- 
               
               
                   
                 ethyl]-methyl-amino}-benzyl)-piperazin-1-yl]-2-oxo- 
               
               
                   
                 ethyl}-3-(4-difluoromethoxy-phenyl)-acrylamide 
               
               
                 15 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1- 
                 1.29 
                 899.32 
               
               
                   
                 benzyl-2-[4-(4-{[2-(tert-butyl-dimethyl-silanyloxy)- 
               
               
                   
                 ethyl]-methyl-amino}-benzyl)-piperazin-1-yl]-2-oxo- 
               
               
                   
                 ethyl}-3-(4-isopropyl-phenyl)-acrylamide 
               
               
                 16 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-{1- 
                 1.32 
                 913.29 
               
               
                   
                 benzyl-2-[4-(4-{[2-(tert-butyl-dimethyl-silanyloxy)- 
               
               
                   
                 ethyl]-methyl-amino}-benzyl)-piperazin-1-yl]-2-oxo- 
               
               
                   
                 ethyl}-3-(4-tert-butyl-phenyl)-acrylamide 
               
               
                   
               
               
                 *Analytic I, X-Bridge column, basic conditions. 
               
             
          
         
       
     
       General Method F—Step 6 
       [0213]    
       
                 
         
             
             
         
       
     
         [0214]    To a solution of 1 mmol of protected alcohol 28 in 3.5 mL of dry THF under nitrogen at 0° C. are added 3.5 mmol of TBAF (1 M solution in THF). The resulting mixture is stirred at RT for 4 h and cooled down to 0° C. Upon completion H 2 O is added and the mixture extracted with DCM (×3). The combined organic phases are washed with brine, dried over Na 2 SO 4 , filtered and concentrated under reduced pressure. The residue is purified either by flash chromatography (SiO 2  60F) or by preparative HPLC to afford the final compound. 
         [0000]    
       
         
               
               
               
             
               
               
               
               
             
           
               
                   
               
               
                 Compound of 
                   
                 LC-MS* 
               
             
          
           
               
                 Example # 
                 Chemical name 
                 Rt (min) 
                 [M + H] +   
               
               
                   
               
               
                 50 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-[1- 
                 0.91 
                 811.21 
               
               
                   
                 benzyl-2-(4-{4-[(2-hydroxy-ethyl)-methyl-amino]- 
               
               
                   
                 benzyl}-piperazin-1-yl)-2-oxo-ethyl]-3-(4- 
               
               
                   
                 trifluoromethyl-phenyl)-acrylamide 
               
               
                 51 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-[1- 
                 0.86 
                 812.24 
               
               
                   
                 benzyl-2-(4-{4-[(2-hydroxy-ethyl)-methyl-amino]- 
               
               
                   
                 benzyl}-piperazin-1-yl)-2-oxo-ethyl]-3-(6- 
               
               
                   
                 trifluoromethyl-pyridin-3-yl)-acrylamide 
               
               
                 52 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-[1- 
                 0.82 
                 774.25 
               
               
                   
                 benzyl-2-(4-{4-[(2-hydroxy-ethyl)-methyl-amino]- 
               
               
                   
                 benzyl}-piperazin-1-yl)-2-oxo-ethyl]-3-(6- 
               
               
                   
                 methoxy-pyridin-3-yl)-acrylamide 
               
               
                 53 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-[1- 
                 0.85 
                 788.24 
               
               
                   
                 benzyl-2-(4-{4-[(2-hydroxy-ethyl)-methyl-amino]- 
               
               
                   
                 benzyl}-piperazin-1-yl)-2-oxo-ethyl]-3-(6-ethoxy- 
               
               
                   
                 pyridin-3-yl)-acrylamide 
               
               
                 54 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-[1- 
                 0.88 
                 787.19 
               
               
                   
                 benzyl-2-(4-{4-[(2-hydroxy-ethyl)-methyl-amino]- 
               
               
                   
                 benzyl}-piperazin-1-yl)-2-oxo-ethyl]-3-(4-ethoxy- 
               
               
                   
                 phenyl)-acrylamide 
               
               
                 55 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-[1- 
                 0.80 
                 821.18 
               
               
                   
                 benzyl-2-(4-{4-[(2-hydroxy-ethyl)-methyl-amino]- 
               
               
                   
                 benzyl}-piperazin-1-yl)-2-oxo-ethyl]-3-(4- 
               
               
                   
                 methanesulfonyl-phenyl)-acrylamide 
               
               
                 56 
                 (S)-3-(4-Acetylamino-phenyl)-N-[4-(4-acetyl- 
                 0.77 
                 800.21 
               
               
                   
                 piperazin-1-yl)-benzyl]-N-[1-benzyl-2-(4-{4-[(2- 
               
               
                   
                 hydroxy-ethyl)-methyl-amino]-benzyl}-piperazin- 
               
               
                   
                 1-yl)-2-oxo-ethyl]-acrylamide 
               
               
                 57 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-[1- 
                 0.92 
                 801.22 
               
               
                   
                 benzyl-2-(4-{4-[(2-hydroxy-ethyl)-methyl-amino]- 
               
               
                   
                 benzyl}-piperazin-1-yl)-2-oxo-ethyl]-3-(4-propoxy- 
               
               
                   
                 phenyl)-acrylamide 
               
               
                 58 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-[1- 
                 0.91 
                 801.25 
               
               
                   
                 benzyl-2-(4-{4-[(2-hydroxy-ethyl)-methyl-amino]- 
               
               
                   
                 benzyl}-piperazin-1-yl)-2-oxo-ethyl]-3-(4- 
               
               
                   
                 isopropoxy-phenyl)-acrylamide 
               
               
                 59 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-[1- 
                 0.92 
                 771.29 
               
               
                   
                 benzyl-2-(4-{4-[(2-hydroxy-ethyl)-methyl-amino]- 
               
               
                   
                 benzyl}-piperazin-1-yl)-2-oxo-ethyl]-3-(4-ethyl- 
               
               
                   
                 phenyl)-acrylamide 
               
               
                 60 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-[1- 
                 0.87 
                 808.87 
               
               
                   
                 benzyl-2-(4-{4-[(2-hydroxy-ethyl)-methyl-amino]- 
               
               
                   
                 benzyl}-piperazin-1-yl)-2-oxo-ethyl]-3-(4- 
               
               
                   
                 difluoromethoxy-phenyl)-acrylamide 
               
               
                 61 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-[1- 
                 0.94 
                 787.34 
               
               
                   
                 benzyl-2-(4-{4-[(2-hydroxy-ethyl)-methyl-amino]- 
               
               
                   
                 benzyl}-piperazin-1-yl)-2-oxo-ethyl]-3-(4- 
               
               
                   
                 isopropyl-phenyl)-acrylamide 
               
               
                 62 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N-[1- 
                 0.97 
                 799.22 
               
               
                   
                 benzyl-2-(4-{4-[(2-hydroxy-ethyl)-methyl-amino]- 
               
               
                   
                 benzyl}-piperazin-1-yl)-2-oxo-ethyl]-3-(4-tert- 
               
               
                   
                 butyl-phenyl)-acrylamide 
               
               
                   
               
               
                 *Analytic I, X-Bridge column, basic conditions. 
               
             
          
         
       
     
       Preparation of Compounds of formula I Via Pathway Depicted in Scheme 3 
     General Procedures and Examples 
     Step 1: (S)-2-[4-(4-Acetyl-piperazin-1-yl)-benzylamino]-3-phenyl-propionic acid methyl ester 
       [0215]    
       
                 
         
             
             
         
       
     
         [0216]    The secondary amine 18 is obtained according to general Methods C1 or C2 and used in the next step without further purification. Rt=0.82; [M+H] + =396.20 (Analytic I, X-Bridge column, basic conditions). 
       Step 2 
       [0217]    
       
                 
         
             
             
         
       
     
         [0218]    The compounds of formula 20 are obtained according to general Methods D1 or D2. 
         [0000]    
       
         
               
               
             
               
               
               
               
               
               
             
           
               
                   
                   
               
               
                   
                 LC-MS 
               
             
          
           
               
                 Intermediate # 
                 Chemical name 
                 Yield 
                 t R  (min) 
                 [M + H] +   
                 conditions 
               
               
                   
               
               
                 17 
                 (S)-2-{[4-(4-Acetyl-piperazin-1- 
                 70% 
                 0.96 
                 594.15 
                 Analytic I, 
               
               
                   
                 yl)-benzyl]-[3-(4-trifluoromethyl- 
                   
                   
                   
                 X-Bridge 
               
               
                   
                 phenyl)-acryloyl]-amino}-3- 
                   
                   
                   
                 column, 
               
               
                   
                 phenyl-propionic acid methyl 
                   
                   
                   
                 basic 
               
               
                   
                 ester 
                   
                   
                   
                 conditions 
               
               
                   
               
             
          
         
       
     
       General Method G—Step 3 
       [0219]    
       
                 
         
             
             
         
       
     
         [0220]    To a solution of 1 mmol of methyl ester 20 in 8 mL of Et 2 O and 2 mL of H 2 O at 0° C. are added dropwise 10 mmol of a 2M aqueous NaOH. The reaction mixture is further stirred at RT for 2-3 h. Upon completion the aqueous phase is acidified to pH=2-3 with HCl 1N and then extracted with EtOAc (×3). The combined organic phases are washed with brine, dried over Na 2 SO 4 , filtered and concentrated under reduced pressure. The residue is used in the next step without further purification. 
         [0000]    
       
         
               
               
             
               
               
               
               
               
               
             
           
               
                   
                   
               
               
                   
                 LC-MS 
               
             
          
           
               
                   
                   
                   
                 t R   
                   
                   
               
               
                 Intermediate # 
                 Chemical name 
                 Yield 
                 (min) 
                 [M + H] +   
                 conditions 
               
               
                   
               
               
                 18 
                 (S)-2-{[4-(4-Acetyl-piperazin-1- 
                 78% 
                 0.97 
                 580.12 
                 Analytic I, 
               
               
                   
                 yl)-benzyl]-[3-(4-trifluoromethyl- 
                   
                   
                   
                 X-Bridge 
               
               
                   
                 phenyl)-acryloyl]-amino}-3- 
                   
                   
                   
                 column, 
               
               
                   
                 phenyl-propionic acid 
                   
                   
                   
                 basic 
               
               
                   
                   
                   
                   
                   
                 conditions 
               
               
                   
               
             
          
         
       
     
       Step 4 
       [0221]    
       
                 
         
             
             
         
       
     
         [0222]    The compounds of formula 8 are obtained according to general Method A. The residue is purified either by flash chromatography (SiO 2  60F) or by preparative HPLC to afford the final compound. 
         [0000]    
       
         
               
               
               
             
               
               
               
               
             
           
               
                   
               
               
                 Compound of 
                   
                 LC-MS* 
               
             
          
           
               
                 Example # 
                 Chemical name 
                 Rt (min) 
                 [M + H] +   
               
               
                   
               
               
                 63 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N- 
                 0.99 
                 768.24 
               
               
                   
                 {1-benzyl-2-[4-(4-methoxy-benzyl)-piperazin- 
               
               
                   
                 1-yl]-2-oxo-ethyl}-3-(4-trifluoromethyl-phenyl)- 
               
               
                   
                 acrylamide 
               
               
                 64 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N- 
                 0.99 
                 783.22 
               
               
                   
                 {1-benzyl-2-[4-(4-nitro-benzyl)-piperazin-1-yl]- 
               
               
                   
                 2-oxo-ethyl}-3-(4-trifluoromethyl-phenyl)- 
               
               
                   
                 acrylamide 
               
               
                 65 
                 (S)-N-[4-(4-Acetyl-piperazin-1-yl)-benzyl]-N- 
                 1.02 
                 796.24 
               
               
                   
                 {1-benzyl-2-[4-(4-isopropoxy-benzyl)- 
               
               
                   
                 piperazin-1-yl]-2-oxo-ethyl}-3-(4- 
               
               
                   
                 trifluoromethyl-phenyl)-acrylamide 
               
               
                   
               
               
                 *Analytic I, X-Bridge column, basic conditions. 
               
             
          
         
       
     
         [0223]    The compounds of Examples 66 to 73 are obtained according to general Method D2. 
         [0000]                                                      Compound of       LC-MS*            Example #   Chemical name   Rt (min)   [M + H] +                 66   (S)-N-[4-(4-Acetyl-piperazin-1-yl)benzyl]-N-{1-(4-   0.94   713.13           (4-(dimethylamino)benzyl)piperazin-1-yl)-1-oxo-           3-phenylpropan-2-yl}cinnamamide       67   (S)-N-[4-(4-Acetyl-piperazin-1-yl)benzyl]-N-{1-[4-   0.97   727.45           (4-(dimethylamino)benzyl)piperazin-1-yl]-1-oxo-3-           phenylpropan-2-yl}-3-(p-tolyl)acrylamide       68   (S)-N-[4-(4-Acetyl-piperazin-1-yl)benzyl]-N-{1-(4-   1.04   755.45           (4-(dimethylamino)benzyl)piperazin-1-yl)-1-oxo-           3-phenylpropan-2-yl}-3-(4-           propylphenyl)acrylamide       69   (S)-N-[4-(4-Acetyl-piperazin-1-yl)benzyl]-N-{1-(4-   0.93   743.46           (4-(dimethylamino)benzyl)piperazin-1-yl)-1-oxo-           3-phenylpropan-2-yl}-3-(4-           methoxyphenyl)acrylamide       70   (S)-3-(4-Acetyl-phenyl)-N-[4-(4-acetyl-piperazin-   0.91   755.43           1-yl)benzyl]-N-{1-(4-(4-           (dimethylamino)benzyl)piperazin-1-yl)-1-oxo-3-           phenylpropan-2-yl}acrylamide       71   (S)-N-[4-(4-Acetyl-piperazin-1-yl)benzyl]-3-(4-   0.92   738.44           cyanophenyl)-N-{1-(4-(4-           (dimethylamino)benzyl)piperazin-1-yl)-1-oxo-3-           phenylpropan-2-yl}acrylamide       72   (S)-N-[4-(4-Acetyl-piperazin-1-yl)benzyl]-N-{1-(4-   0.95   731.42           (4-(dimethylamino)benzyl)piperazin-1-yl)-1-oxo-           3-phenylpropan-2-yl}-3-(4-           fluorophenyl)acrylamide       73   (S)-N-[4-(4-Acetyl-piperazin-1-yl)benzyl]-3-(4-   0.98   747.28           chlorophenyl)-N-{1-(4-(4-           (dimethylamino)benzyl)piperazin-1-yl)-1-oxo-3-           phenylpropan-2-yl}acrylamide               *Analytic I, X-Bridge column, basic conditions.            
In Vitro Antimalarial Activity:  Plasmodium falciparum  In Vitro Assay
 
         [0224]    In vitro activity against erythrocytic stages of  P. falciparum  in human red blood cells is determined using a [ 3 H] hypoxanthine incorporation assay. One strain sensitive to all known drugs ( P. falciparum  NF54) is used in this assay and all tested compounds are compared for activity with the standard drugs chloroquine (sigma C6628) and artesunate (sigma 36, 159-3). Compounds, tested in duplicates, are serially diluted with screening medium [RPMI 1640 medium, supplemented with HEPES (5.94 g/L), NaHCO 3  (2.1 g/L), neomycin (100 U/mL), and Albumax (5 g/L) or human serum (50% final concentration)] in 96-well microtiter plates within an appropriate concentration range. Thereafter, the parasite cultures incubated in screening medium containing washed human red blood cells at 2.5% hematocrit (0.3% parasitemia) are added to the serially diluted compounds and incubated in a humidifying atmosphere at 37° C., 4% CO 2 , 3% O 2 , and 93% N 2 . After 48 h, [ 3 H] hypoxanthine (0.5 μCi) is added to each well of a plate. The plates are incubated for a further 24 h under the same conditions then harvested with a Betaplate cell harvester (Wallac) and washed with distilled water. The dried filters are inserted into a plastic foil with 10 mL of scintillation fluid, and counted in a Betaplate liquid scintillation counter. IC 50  values are calculated from sigmoidal inhibition curves using Microsoft Excel. 
         [0000]    
       
         
               
             
               
               
               
             
               
               
               
             
           
               
                 TABLE 1 
               
             
             
               
                   
               
               
                 IC 50  values (nM) for compounds of formula I: 
               
             
          
           
               
                   
                 IC 50  on NF54 
                 IC 50  on NF54 
               
               
                 Compound of Example # 
                 (with Albumax) 
                 (with 50% Serum) 
               
               
                   
               
             
          
           
               
                  1 
                 5.1 
                 27.6 
               
               
                 (Reference Example) 
               
               
                  2 
                 2.0 
                 10 
               
               
                  3 
                 1.9 
                 8.5 
               
               
                  4 
                 1.9 
                 10 
               
               
                  5 
                 3.9 
                 12 
               
               
                  6 
                 9.4 
                 32 
               
               
                  7 
                 3.1 
                 40 
               
               
                  8 
                 1.8 
                 13.4 
               
               
                  9 
                 2.4 
                 7.3 
               
               
                 10 
                 3.0 
                 8.6 
               
               
                 11 
                 4.0 
                 21 
               
               
                 12 
                 17 
                 74 
               
               
                 13 
                 12 
                 46 
               
               
                 14 
                 3.7 
                 20 
               
               
                 15 
                 1.7 
                 6.8 
               
               
                 16 
                 5.5 
                 13.3 
               
               
                 17 
                 4.3 
                 39 
               
               
                 18 
                 2.5 
                 11.8 
               
               
                 19 
                 2.6 
                 9.3 
               
               
                 20 
                 1.2 
                 7.1 
               
               
                 21 
                 1.9 
                 7 
               
               
                 22 
                 1.3 
                 10.9 
               
               
                 23 
                 1.3 
                 9.5 
               
               
                 24 
                 2.8 
                 13.9 
               
               
                 25 
                 1.2 
                 4.7 
               
               
                 26 
                 18 
                 33 
               
               
                 27 
                 1.8 
                 12.9 
               
               
                 28 
                 1.1 
                 3.6 
               
               
                 29 
                 1.1 
                 3.7 
               
               
                 30 
                 0.9 
                 5.0 
               
               
                 31 
                 1.7 
                 3.8 
               
               
                 32 
                 0.5 
                 4.4 
               
               
                 33 
                 3.3 
                 10.9 
               
               
                 34 
                 0.5 
                 3.8 
               
               
                 35 
                 2.9 
                 9 
               
               
                 36 
                 7.8 
                 28 
               
               
                 37 
                 3.2 
                 28 
               
               
                 38 
                 1.8 
                 9 
               
               
                 39 
                 2.3 
                 11 
               
               
                 40 
                 1.7 
                 10.5 
               
               
                 41 
                 2.9 
                 12.5 
               
               
                 42 
                 1.7 
                 13.7 
               
               
                 43 
                 1.1 
                 13.5 
               
               
                 44 
                 0.4 
                 2.8 
               
               
                 45 
                 0.2 
                 1.8 
               
               
                 46 
                 0.4 
                 1.1 
               
               
                 47 
                 0.5 
                 3.5 
               
               
                 48 
                 0.5 
                 1.6 
               
               
                 49 
                 0.8 
                 3.1 
               
               
                 50 
                 1.6 
                 9.3 
               
               
                 51 
                 10.7 
                 29.8 
               
               
                 52 
                 10 
                 33 
               
               
                 53 
                 4.3 
                 12.4 
               
               
                 54 
                 1.6 
                 5.9 
               
               
                 55 
                 7.5 
                 14.8 
               
               
                 57 
                 1.7 
                 5.1 
               
               
                 58 
                 1.4 
                 3.9 
               
               
                 59 
                 1.6 
                 3.8 
               
               
                 60 
                 3 
                 8.1 
               
               
                 61 
                 1.2 
                 6.7 
               
               
                 62 
                 0.7 
                 3.2 
               
               
                 63 
                 3.2 
                 17.8 
               
               
                 64 
                 3.5 
                 43 
               
               
                 65 
                 2.8 
                 14 
               
               
                 66 
                 24.6 
                 113 
               
               
                 67 
                 &lt;3.1 
                 17.5 
               
               
                 68 
                 0.7 
                 8 
               
               
                 69 
                 &lt;3.1 
                 17 
               
               
                 70 
                 &lt;3.1 
                 16 
               
               
                 71 
                 18 
                 62 
               
               
                 72 
                 10 
                 91 
               
               
                 73 
                 3.3 
                 26 
               
               
                 Chloroquine 
                 4.2 
                 3.8 
               
               
                 Artesunate 
                 2.7 
                 1 
               
               
                   
               
             
          
         
       
     
       In Vivo Antimalarial Efficacy Studies 
       [0225]    In vivo antimalarial activity is assessed for groups of three female NMRI mice (20-22 g) intravenously infected on day 0 with  P. berghei  strain GFP-ANKA (0.2 mL heparinized saline suspension containing 2×10 7  parasitized erythrocytes). In control mice, parasitemia typically rises to approximately 40% by day 3 after infection. Compounds are formulated in Tween 80/ethanol (7%/3%) usually at concentrations of 10 mg/mL. Compounds are administered in a volume of 10 mL/kg orally as single doses (1×100 mg/kg, 24 h after infection). 48 h after drug treatment (day 3 post-infection), 1 μl tail blood is taken, resuspended in 1 mL PBS buffer and parasitemia determined with a FACScan (Becton Dickinson) by counting 100′000 red blood cells. Activity is calculated as the difference between the mean value of the control group and treated groups expressed as a percent relative to the control group. 
         [0000]    
       
         
               
             
               
               
               
             
               
               
               
             
           
               
                 TABLE 2 
               
             
             
               
                   
               
               
                 Activity (inhibition of parasitemia) of compounds of formula I after single 
               
               
                 oral dose of 100 mg/kg expressed as a percent relative to the control group 
               
             
          
           
               
                   
                 Compound of 
                   
               
               
                   
                 Example # 
                 Activity [%] 
               
               
                   
                   
               
             
          
           
               
                   
                  1* 
                 &lt;40 
               
               
                   
                  2 
                 94.1 
               
               
                   
                  3 
                 98.7 
               
               
                   
                  4 
                 97.4 
               
               
                   
                  5 
                 &lt;40 
               
               
                   
                  6 
                 80.4 
               
               
                   
                  7 
                 44.7 
               
               
                   
                  8 
                 99.4 
               
               
                   
                  9 
                 99.1 
               
               
                   
                 10 
                 96 
               
               
                   
                 15 
                 99 
               
               
                   
                 16 
                 85.9 
               
               
                   
                 18 
                 99.1 
               
               
                   
                 19 
                 99.2 
               
               
                   
                 20 
                 98.5 
               
               
                   
                 21 
                 98.8 
               
               
                   
                 22 
                 99.3 
               
               
                   
                 23 
                 99.1 
               
               
                   
                 24 
                 79.2 
               
               
                   
                 25 
                 99.1 
               
               
                   
                 27 
                 &lt;40 
               
               
                   
                 28 
                 99.3 
               
               
                   
                 29 
                 99.2 
               
               
                   
                 30 
                 97.7 
               
               
                   
                 31 
                 88.9 
               
               
                   
                 32 
                 98.5 
               
               
                   
                 33 
                 97.4 
               
               
                   
                 34 
                 99.2 
               
               
                   
                 35 
                 98.7 
               
               
                   
                 36 
                 46.8 
               
               
                   
                 38 
                 98.5 
               
               
                   
                 39 
                 99 
               
               
                   
                 40 
                 93.6 
               
               
                   
                 41 
                 71 
               
               
                   
                 42 
                 97.3 
               
               
                   
                 43 
                 98.7 
               
               
                   
                 44 
                 99.0 
               
               
                   
                 45 
                 99.4 
               
               
                   
                 46 
                 98.9 
               
               
                   
                 47 
                 98.4 
               
               
                   
                 48 
                 99 
               
               
                   
                 49 
                 98.6 
               
               
                   
                 50 
                 98 
               
               
                   
                 52 
                 &lt;40 
               
               
                   
                 53 
                 52.3 
               
               
                   
                 54 
                 93.7 
               
               
                   
                 55 
                 &lt;40 
               
               
                   
                 57 
                 98.2 
               
               
                   
                 58 
                 98.2 
               
               
                   
                 59 
                 96.5 
               
               
                   
                 60 
                 78.3 
               
               
                   
                 61 
                 98.8 
               
               
                   
                 62 
                 99.4 
               
               
                   
                   
               
               
                   
                 *Reference Example