In Vivo Methods for Enhancing Bioenergetic Status in Female Germ Cells

Abstract

In vivo methods using bioenergetic agents for restoring the quality of aged oocytes, enhancing oogonial stem cells or improving derivatives thereof (e.g., cytoplasm or isolated mitochondria) for use in fertility-enhancing procedures, are described.

1 . A method of improving fertility in a female mammalian subject in need thereof, said method comprising administering to the female mammalian subject one or more bioenergetics agents in an amount effective to enhance the bioenergetic status of an oocyte or an oogonial stem cell (OSC) of the female mammalian subject, thereby improving fertility in the female mammalian subject. 
     
     
         2 . The method of  claim 1 , wherein the bioenergetic agent is selected from the group consisting of a CD38 inhibitor, a nicotinamide adenine dinucleotide (NAD) precursor and combinations thereof. 
     
     
         3 . The method of  claim 2 , wherein the CD38 inhibitor is a compound selected from the group consisting of 1-[(2-Acetoxyethoxy)methyl]-3-(aminocarbonyl)-pyridinium chloride, 1-[(2-Benzyloxyethoxy)methyl]-3-(aminocarbonyl)-pyridinium chloride, 1{[2-(4-Methoxy-phenoxy)ethoxy]methyl}-3-(aminocarbonyl)-pyridinium chloride, 1-{[2-(4-Phenoxy-phenoxy)ethoxy]methyl}-3-(aminocarbonyl)-pyridinium chloride, 1-{[2-(4-Nitro-phenoxy)ethoxy]methyl}-3-(aminocarbonyl)-pyridinium chloride, 1-{[2-(3-Trifluoromethyl-phenoxy)ethoxy]methyl}-3-(aminocarbonyl)-pyridinium chloride, 1-{[2-(8′-Quinolyloxy)ethoxy]methyl}-3-(aminocarbonyl)-pyridinium chloride, 1,2-Dimethoxy-ethylene-bis-N,N′-3-(aminocarbonyl)-pyridinium dichloride, 1,4-Dimethoxy-butylene-bis-N,N′-3-(aminocarbonyl)-pyridinium dichloride, 1,4-Dimethoxy-butyne-bis-N,N′-3-(aminocarbonyl)-pyridinium dichloride, 1,4-Dimethoxy-hexamethylene-bis-N,N′-3-(aminocarbonyl)-pyridinium dichloride, (E)-1-{[4-(8′-Quinolyloxy)but-2-enyloxy]methyl}-3-(aminocarbonyl)-pyridinium chloride, 1-{[2-(4-Phenoxy-phenoxy)ethoxy]methyl}-6-(aminocarbonyl)-quinolinium chloride, 1-{[2-(4-Phenoxy-phenoxy)ethoxy]methyl}-3-(aminocarbonyl)-4-amino-pyridinium chloride, luteolinidin, kuromanin, luteolin, delphinidin, pelargonidin, malvidin, quercetagetinidin, peonidin, myricetin, cyanidin, diosmetinidin, quercetin, robinetin, petunidin, fisetin, fisetinidin, quercetagetin, rac-taxifolin, rac-catechin, piceatannol, resveratrol, apigenin, tyrphostin-8, berberine and SRT-1720. 
     
     
         4 . The method of  claim 2 , wherein the NAD precursor is a compound selected from the group consisting of nicotinamide mononucleotide, nicotinamide riboside and nicotinic acid. 
     
     
         5 . The method of  claim 2 , wherein the bioenergetic agent comprises a CD38 inhibitor and an NAD precursor. 
     
     
         6 . The method of  claim 3 , wherein the CD38 inhibitor is a compound selected from the group consisting of apigenin, luteolin, tyrphostin-8, berberine and SRT-1720, and
 the NAD precursor is a compound selected from the group consisting of nicotinamide mononucleotide, nicotinamide riboside and nicotinic acid.   
     
     
         7 . The method of  claim 1 , wherein the one or more bioenergetic agents are administered systemically to the female mammalian subject. 
     
     
         8 . The method of  claim 1 , wherein the one or more bioenergetic agents are administered locally to an ovary of the female mammalian subject. 
     
     
         9 . The method of  claim 1 , wherein the one or more bioenergetic agents are administered in a composition comprising a therapeutically effective amount of the one or more bioenergetic agents and a pharmaceutically acceptable carrier. 
     
     
         10 . The method of  claim 1 , wherein the pregnancy outcome of the female mammalian subject is improved compared to a reference standard. 
     
     
         11 . The method of  claim 1 , wherein the OSC is a non-embryonic stem cell, and is mitotically competent and expresses Vasa, Oct-4, Dazl and Stella and, optionally, a stage-specific embryonic antigen. 
     
     
         12 . The method of  claim 4 , wherein the NAD precursor is nicotinamide riboside. 
     
     
         13 . The method of  claim 4 , wherein the NAD precursor is nicotinic acid. 
     
     
         14 . The method of  claim 1 , wherein the female mammalian subject is a human female selected from the group consisting of females of advanced maternal age, females suffering from oocyte-related infertility, females with low ovarian reserve, females suffering from premature ovarian failure and females suffering from infertility due to chemotherapy. 
     
     
         15 . The method of  claim 1 , wherein the one or more bioenergetic agents are administered in an amount effective to increase the number of functional mitochondria in the oocyte or OSC of the female mammalian subject. 
     
     
         16 . The method of  claim 1 , wherein the one or more bioenergetic agents are administered in an amount effective to increase the mitochondrial energy of the oocyte or the OSC of the female mammalian subject. 
     
     
         17 . The method of  claim 1 , wherein the one or more bioenergetic agents are administered in an amount effective to increase the cellular energy of the oocyte or OSC of the female mammalian subject. 
     
     
         18 . The method of  claim 1 , wherein the one or more bioenergetic agents are administered in an amount effective to improve oocyte quality, OSC quality, de novo production or ovulated oocyte yield. 
     
     
         19 . The method of  claim 9 , wherein the CD38 inhibitor is present in the composition at a concentration ≥25 μM. 
     
     
         20 . The method of  claim 9 , wherein the NAD precursor is present in the composition at a concentration ≥100 μM. 
     
     
         21 . The method of  claim 11 , wherein the OSC expresses a stage-specific embryonic antigen. 
     
     
         22 . A method of in vitro fertilization, said method comprising the steps of:
 a) administering to a female mammalian subject one or more bioenergetic agents in an amount effective to enhance the bioenergetic status of an oocyte or an oogonial stem cell (OSC) of the female mammalian subject;   b) obtaining an oocyte from the female mammalian subject; and   c) fertilizing the oocyte in vitro to form a zygote.   
     
     
         23 - 48 . (canceled) 
     
     
         49 . A method of sustaining embryonic development in a pregnant female subject in need thereof, said method comprising administering to the subject a therapeutically effective amount of one or more bioenergetic agents in an amount effective to enhance the bioenergetic status of an embryo of the female mammalian subject, thereby sustaining embryonic development in the pregnant female subject. 
     
     
         50 - 51 . (canceled) 
     
     
         52 . A method of restoring or increasing ovarian function in a female mammalian subject in need thereof, comprising administering a therapeutically effective amount of one or more bioenergetics agents in an amount effective to enhance the bioenergetic status of an oocyte or an oogonial stem cell (OSC) of the female mammalian subject, thereby restoring ovarian function in the female mammalian subject. 
     
     
         53 - 54 . (canceled) 
     
     
         55 . A method of preparing a tissue or cell thereof from a female mammalian subject for harvest, comprising administering to the female mammalian subject one or more bioenergetics agents in an amount effective to enhance the bioenergetic status the tissue or cell thereof, thereby preparing the tissue or cell thereof from the female mammalian subject for harvest. 
     
     
         56 - 58 . (canceled)