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Small-Molecule Inhibition of BRDT for Male Contraception
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SUMMARY
A pharmacologic approach to male contraception remains a longstanding challenge in medicine. Toward this objective, we explored the spermatogenic effects of a selective small-molecule inhibitor (JQ1) of the bromodomain and extraterminal (BET) subfamily of epigenetic reader proteins. Here, we report potent inhibition of the testis-specific member BRDT, which is essential for chromatin remodeling during spermatogenesis. Biochemical and crystallographic studies confirm that occupancy of the BRDT acetyl-lysine binding pocket by JQ1 prevents recognition of acetylated histone H4. Treatment of mice with JQ1 reduced seminiferous tubule area, testis size, and spermatozoa number and motility without affecting hormone levels. Although JQ1-treated males mate normally, inhibitory effects of JQ1 evident at the spermatocyte and round spermatid stages cause a complete and reversible contraceptive effect. These data establish a new contraceptive that can cross the blood:testis boundary and inhibit
bromodomain activity during spermatogenesis, providing a lead compound targeting the male germ cell for contraception.

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A pharmacologic approach to male contraception remains a longstanding challenge in medicine. Toward this objective, we explored the spermatogenic effects of a selective small-molecule inhibitor (JQ1) of the bromodomain and extraterminal (BET) subfamily of epigenetic reader proteins.
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Here, we report potent inhibition of the testis-specific member BRDT, which is essential for chromatin remodeling during spermatogenesis.
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Biochemical and crystallographic studies confirm that occupancy of the BRDT acetyl-lysine binding pocket by JQ1 prevents recognition of acetylated histone H4.


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Treatment of mice with JQ1 reduced seminiferous tubule area, testis size, and spermatozoa number and motility without affecting hormone levels.
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These data establish a new contraceptive that can cross the blood:testis boundary and inhibit bromodomain activity during spermatogenesis, providing a lead compound targeting the male germ cell for contraception.

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Although ~4% of the mammalian genome encodes genes expressed in male germ cells during spermatogenesis (Schultz et al., 2003), contraceptive drugs for men have remained elusive. To date, the only drugs in clinical trials are testosterone analogs
that alter endogenous androgen production, although there is a short list of other possible targets (e.g., GAPDHS) and drugs (e.g., gamendazole) (Aitken et al., 2008). This lack of contraceptive alternatives for men is partially responsible for the high rate
of unplanned pregnancies, especially in teenagers, and contributes to the maternal mortality, ethical, social, and financial costs associated with abortions and deliveries to single mothers. To approach this dearth of contraceptive alternatives for men, we
have undertaken to develop small molecules that could target spermatogenic-specific proteins that have been shown to be essential for both spermatogenesis and fertility in mammals. One such contraceptive target is the testis-specific and bromodomain-
containing protein BRDT.

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Although ~4% of the mammalian genome encodes genes expressed in male germ cells during spermatogenesis (Schultz et al., 2003), contraceptive drugs for men have remained elusive.
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cell for contraception. ÕªÒª²¿·Ö×îºóÒ»¾ä£©¡£

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To date, the only drugs in clinical trials are testosterone analogs
that alter endogenous androgen production, although there is a short list of other possible targets (e.g., GAPDHS) and drugs (e.g., gamendazole) (Aitken et al., 2008).

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This lack of contraceptive alternatives for men is partially responsible for the high rate
of unplanned pregnancies, especially in teenagers, and contributes to the maternal mortality, ethical, social, and financial costs associated with abortions and deliveries to single mothers.

This lack ofÕâÖÖÓ÷¨Ò²ÊǺܳ£¼ûµÄ£¬Äã¾ø¶ÔÓõÃ×Å£¬lackÕâ¸ö´Ê±È½ÏÆæ¹Ö£¬´ó¼Ò¿ÉÒÔ×Ðϸ²é²é¡£´ËÍâis  responsible for ºÍcontributes toµÄÓ÷¨Ò²ºÜ³£¼û¡£¼Çס¸±´Êpartially¡¢especiallyµÄÓ÷¨£¬³õѧÕßÒªÌØ±ð×¢Ò⸱´ÊºÍÁ¬´ÊµÄÓ÷¨¡£
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To approach this dearth of contraceptive alternatives for men, we
have undertaken to develop small molecules that could target spermatogenic-specific proteins that have been shown to be essential for both spermatogenesis and fertility in mammals. One such contraceptive target is the testis-specific and bromodomain-
containing protein BRDT.

To approach this dearth of contraceptive alternatives for men, ΪÁ˽â¾öȱ·¦ÄÐÐÔ±ÜÔз½·¨µÄÀ§¾³£¬To approach£¬ÖµµÃÄóöС±Ê¼Ç±¾¼ÇÏÂÀ´¡£
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Genetic studies of BRDT have demonstrated that selective deletion of the BRDT(1)-encoding region is sufficient to confer sterility in homozygous hypomorphic male mice (Shang et al., 2007), and a recently published genome-wide association study of idiopathic male infertility identified single-nucleotide polymorphisms of BRDT as significantly associated with oligozoospermia or azoospermia in European men (Aston et al., 2010). These insights establish a compelling rationale to target BRDT for a contraceptive effect.
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These insights establish a compelling rationale to target BRDT for a contraceptive effect.
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Recently, we established the feasibility of targeting human bromodomains with acetyl-lysine competitive small molecules (Filippakopoulos et al., 2010).
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complete and reversible contraceptive effect in males without adversely affecting testosterone levels or mating behaviors and without prompting obvious teratogenic effects in offspring.
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These results indicate that targeting a developmental epigenetic reader protein with
an orally bioavailable small molecule can modulate male fertility for a contraceptive effect.

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JQ1 Selectively Targets Male Germ Cells without
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in JQ1-Treated Testes
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