Method and pharmacological composition for the diagnosis and treatment of male

Abstract

Pharmaceutical compositions for treating male sub-fertility that include an agent that causes a reduction in an effect of extracellular DNA on sperm cells, are provided. The agent may be, for example, an enzyme that degrades DNA such as DNase, a substance that blocks the interaction between cell free DNA and sperm cell surface receptors, a substance that binds to DNA, a substance that inhibits endogenous sperm cell DNase, a substance that inhibits a member of a signal transduction pathway mediated by DNA binding to sperm cell surface receptors, or an agent that stimulates production of an endogenous substance that causes a reduction in an antifertility effect of cell free DNA on sperm cells. Also provided are methods for treating male sub-fertility by administering the pharmaceutical composition to a subject in need thereof. Further provided are methods for determining a fertility status in a male subject, methods for assisted reproduction, methods for selecting an assisted reproduction technique (ART), and methods for selecting sperm cells in a sperm cell population for use in an assisted reproduction technique.

1 - 48 . (canceled) 
     
     
         49 . A pharmaceutical composition for use in the treatment of male sub-fertility comprising:
 an active ingredient selected from the group consisting of: a deoxyribonuclease (DNase), a substance that blocks the interaction between cell free DNA and sperm cell surface receptors, a substance that binds to DNA, a substance that inhibits endogenous sperm cell Dnase, a substance that inhibits a member of a signal transduction pathway mediated by DNA binding to sperm cell surface receptors, an agent that stimulates production of an endogenous substance that causes a reduction in an antifertility effect of cell free DNA on sperm cells, an agent that causes a reduction in an effect of extracellular DNA on sperm cells, and an agent that stimulates production of an endogenous substance that causes a reduction in an effect of extracellular DNA on sperm cells; and   a physiologically acceptable carrier.   
     
     
         50 . The composition according to  claim 49 , wherein the active ingredient is DNase I. 
     
     
         51 . The composition according to  claim 49 , wherein the enzyme is of animal, plant, bacterial, viral, yeast, or protozoan origin or is a recombinant enzyme or a human recombinant enzyme. 
     
     
         52 . The composition according to  claim 49 , in a dosage form suitable for oral administration. 
     
     
         53 . The composition according to  claim 49 , in a dosage form suitable for administration by inhalation. 
     
     
         54 . The composition according to  claim 49 , in a dosage form suitable for administration by injection. 
     
     
         55 . The composition according to  claim 49 , wherein the active ingredient is glycoprotein IF-1. 
     
     
         56 . The pharmaceutical composition according to  claim 49 , wherein the receptor is selected from the group consisting of CD4 and MHC II. 
     
     
         57 . The composition according to  claim 49 , wherein the active ingredient is selected from the group consisting of aurintricarboxylic acid (ATA), citric acid, and a salt thereof. 
     
     
         58 . The composition according to  claim 49 , wherein the active ingredient is a caspase inhibitor. 
     
     
         59 . A method for treating male sub-fertility, comprising administering to an individual in need thereof, a therapeutically effective amount of a pharmaceutical composition according to  claim 49 . 
     
     
         60 . The method according to  claim 59 , wherein the duration of the administration is acute, semi-chronic, or chronic. 
     
     
         61 . A method for determining a fertility status in a male subject, comprising:
 obtaining from the subject a sample of a body tissue or fluid;   measuring in the sample a cell-free DNA level or a level of an agent that causes a reduction in the effect of cell-free DNA on sperm cells;   comparing the measured level to one or more predetermined threshold values; and   determining the fertility status based upon the comparison.   
     
     
         62 . The method according to  claim 61 , wherein the agent is DNase 
     
     
         63 . A method for determining a fertility status in a male subject, comprising:
 obtaining from the subject a sample containing sperm cells;   measuring in the sperm cells a level of a substance indicative of apoptosis;   comparing the measured level to one or more predetermined threshold values; and   determining the fertility status based upon the comparison.   
     
     
         64 . The method according to  claim 63 , wherein the substance is selected from the group consisting of Fas receptor, caspas 3, and sperm cell internal DNase. 
     
     
         65 . A method for assisted reproduction, comprising:
 obtaining a sample containing sperm cells from a male treated by the method of  claim 59 ; and   utilizing the sample in an assisted reproduction technique (ART).   
     
     
         66 . A method for selecting an assisted reproduction technique (ART), comprising:
 determining a fertility status in a male subject by the method of  claim 63 ; and   determining an ART based upon the fertility status.   
     
     
         67 . A method for selecting sperm cells in a sperm cell population for use in an assisted reproduction technique, comprising:
 obtaining a semen sample containing sperm cells; and   removing from the semen sample sperm cells that express an apoptotic marker so as to obtain a sub-population of sperm cells.   
     
     
         68 . The method according to  claim 67 , wherein the marker is cell surface Fas receptor. 
     
     
         69 . The method according to  claim 67 , wherein the marker is the complex DNA-sperm cell surface receptor. 
     
     
         70 . The method according to  claim 67 , wherein the step of removing sperm cells comprises binding of the marker by an antibody; and removing from the semen sample the antibody-bound sperm cells. 
     
     
         71 . The method according to  claim 70 , wherein the antibody is fluorescently labeled and sperm cells bound to the antibodies are removed using fluorescence-activated cell sorting. 
     
     
         72 . The method according to  claim 70 , further comprising binding the antibody to resin beads; and removing resin-bound sperm cells from unbound sperm cells using a column, centrifugation or magnets. 
     
     
         73 . The method according to  claim 65 , wherein the assisted reproduction technique is selected from the group consisting of intra-uterine insemination, in vitro fertilization, intra-cytoplasmic sperm injection, and intra-cytoplasmatic morphologically-selected injection.