Testosterone-inducing peptide compounds and related combinations

Abstract

The present disclosure relates to peptide compounds of formula (I) or (II) and combinations thereof that can be administered orally for promoting endogenous steroid, and particularly testosterone production: A-Xaa1Xaa2-Xaa3-Xaa4-Xaa5-Xaa6-Xaa7-Xaa8-B (I) A-Xaa1-Xaa A -Xaa B -Xaa2-Xaa3-Xaa4-Xaa5-Xaa6-Xaa7-Xaa C -Xaa D -Xaa E -Xaa8-B (II).

Description

[0001] Citations of relevant applications and documents

[0002] This application claims priority to U.S. Provisional Application No. 62 / 756,767, filed on November 7, 2018, which is incorporated herein in its entirety. The sequence listing associated with this application is provided in text format in lieu of a paper copy and is incorporated into this specification by reference. The name of the text file containing the sequence listing is 56080224-2PCT_Sequence listing as submitted. The text file is 12KB, created on November 4, 2019, and submitted electronically via EFS-Web. Technical Field

[0003] The present disclosure relates to a therapeutic compound having an amino acid core capable of inducing endogenous testosterone production. Background Art

[0004] Starting at age 30, testosterone levels in men decrease physiologically at a rate of 0.4 to 2% per year. The hypothalamus-pituitary-gonadal (HPG) axis regulates these testosterone levels through a series of feedback loops. In short, testosterone is produced in the testicles by interstitial cells in response to luteinizing hormone (LH), which is released by gonadotrophic cells of the anterior pituitary. The release of LH is controlled by the release of hypothalamic gonadotropin-releasing hormone (GnRH), which is pulsatile and follows a physiological rhythm.

[0005] Hypogonadism is a reduction in testosterone levels, which is usually accompanied by erectile dysfunction, decreased muscle mass, gynecomastia and osteoporosis, as well as physiological symptoms such as fatigue, mental confusion and decreased libido. Primary hypogonadism occurs when the testicular interstitial cells produce insufficient androgens. However, in most cases (85%), hypogonadism is secondary, in which GnRH or LH signaling is not enough to maintain testosterone levels. There is also a male subgroup with a mixture of central (hypothalamic and / or pituitary) and gonadal deficiency (called late-onset hypogonadism). This group is characterized by reduced testosterone levels, in which symptoms of aging overlap with those of hypogonadism, causing their treatment options to be confused. Other forms of hypogonadism include compensatory hypogonadism, in which LH levels increase to maintain testosterone levels within the low normal range and most commonly develop into primary hypogonadism. Furthermore, testosterone levels may be reduced during trauma and may be a biomarker for surgical outcome.

[0006] Testosterone replacement therapy (TRT) is currently the only approved treatment for hypogonadism, and uses synthetic testosterone analogs to improve patients' symptoms. Side effects of TRT include polycythemia, gynecomastia, and infertility. Other side effects are related to the route of administration, such as cross-contamination in patients using gels and rashes in patients using patches. There are also concerns about inhibiting reproduction and abuse. In addition, retrospective and prospective studies have raised concerns about cardiovascular disease. However, some studies have shown no effect or even no improvement in cardiovascular disease. All these concerns have led the FDA to issue warnings and guidelines aimed at reducing the abuse of TRT, and prescriptions for TRT have tripled in the past decade. Abuse of TRT is also a problem because improper use of TRT may affect the HPG axis for up to 2-3 years, and in some cases permanently. The FDA limits the use of TRT to those individuals with a history of reduced testosterone levels, and excludes those individuals who use TRT to counteract physiological testosterone reduction and those individuals with unknown etiology. The FDA also raises concerns about cardiovascular disease by forcing warning labels on TRT packaging.

[0007] Parenteral administration is an accurate way to dose TRT, but users often miss doses and compliance is minimal. Due to liver toxicity, the oral route, which shows the best compliance, is not a treatment option. New alternatives to TRT that use the oral route and work within the HPG axis are needed. The use of hCG, modulators of estrogen receptors (ER) (such as clomiphene citrate) and aromatase inhibitors (which block the conversion of testosterone into estrogen) have been proposed, but are not currently approved by the FDA. In general, there is an urgent need to redouble efforts to find a viable oral therapy.

[0008] In some embodiments, it would be highly desirable to provide therapeutic agents suitable for oral dosage for treating or alleviating symptoms associated with hypogonadism. Summary of the invention

[0009] The present disclosure relates to peptide compounds and combinations of peptide compounds that are capable of promoting steroid and, in some embodiments, testosterone production when administered orally.

[0010] In a first aspect, the present disclosure provides a peptide compound of formula (I):

[0011] A-Xaa 1 -Xaa 2 -Xaa 3 -Xaa 4 -Xaa 5 -Xaa 6 -Xaa 7 -Xaa8 -B (I)

[0012] wherein: A is present or absent and is a moiety that improves the circulating half-life of the peptide compound; Xaa 1 Exist or not, when present, Xaa 1 is one or more L-amino acid residues or D-amino acid residues; Xaa 2 Exist or not, when present, Xaa 2 is L-serine or D-serine; Xaa 3 is L-lysine, D-lysine, L-arginine or D-arginine; Xaa 4 is L-valine, D-valine, L-isoleucine, D-isoleucine, L-leucine, D-leucine, D-glycine, L-glycine, D-alanine or L-alanine; Xaa 5 is L-serine, D-serine, L-threonine or D-threonine; Xaa 6 is L-glutamine, D-glutamine, L-glutamic acid or D-glutamic acid; Xaa 7 Exist or not, when present, Xaa 7 is L-serine or D-serine; Xaa 8 Exist or not, when present, Xaa 8 Xaa is one or more L-amino acid residues or D-amino acid residues; and B is present or absent and is a moiety that improves the circulating half-life of the peptide compound. 1 In a further embodiment, Xaa 2 In such embodiments, Xaa 3 -Xaa 4 -Xaa 5 -Xaa 6 -Xaa 7 Can have the amino acid sequence of SEQ ID NO:1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 18, 19, 20 or 21. In an alternative embodiment, Xaa 2 In such embodiments, Xaa 2 -Xaa 3 -Xaa 4 -Xaa 5 -Xaa 6 -Xaa 7 It may have the amino acid sequence of SEQ ID NO: 16 or 23. In some embodiments, Xaa 3 is L-lysine or D-lysine. In such embodiments, Xaa 3-Xaa 4 -Xaa 5 -Xaa 6 -Xaa 7 It may have the amino acid sequence of SEQ ID NO: 1, 10, 11, 19 or 20. In some embodiments, Xaa 3 is L-lysine or D-lysine. In such embodiments, Xaa 3 -Xaa 4 -Xaa 5 -Xaa 6 -Xaa 7 It may have the amino acid sequence of SEQ ID NO: 2, 3, 4, 5, 6, 7, 8, 9, 12, 13, 14, 15, 16, 17, 18, 21, 22 or 23. In some embodiments, Xaa 4 is L-valine or D-valine. In such embodiments, Xaa 3 -Xaa 4 -Xaa 5 -Xaa 6 -Xaa 7 It may have an amino acid sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 12, 13, 17, 21, 22 or 23. In some embodiments, Xaa 4 is L-isoleucine or D-isoleucine. In such embodiments, Xaa 3 -Xaa 4 -Xaa 5 -Xaa 6 -Xaa 7 It may have the amino acid sequence of SEQ ID NO: 8, 9, 10, 11, 14, 15, 16, 19 or 20. In some embodiments, Xaa 4 is L-leucine or D-leucine. In such embodiments, Xaa 3 -Xaa 4 -Xaa 5 -Xaa 6 -Xaa 7 Can have the amino acid sequence of SEQ ID NO: 18. In some embodiments, Xaa 5 is L-serine or D-serine. In such embodiments, Xaa 3 -Xaa 4 -Xaa 5 -Xaa 6 -Xaa 7 It may have the amino acid sequence of SEQ ID NO: 1, 5, 6, 7, 16, 19, 20 or 22. In some embodiments, Xaa5 is L-threonine or D-threonine. In such embodiments, Xaa 3 -Xaa 4 -Xaa 5 -Xaa 6 -Xaa 7 It may have an amino acid sequence of SEQ ID NO: 2, 3, 4, 8, 9, 10, 11, 12, 13, 14, 15, 17, 18, 21 or 23. In some embodiments, Xaa 6 is L-glutamine or D-glutamine. In such embodiments, Xaa 3 -Xaa 4 -Xaa 5 -Xaa 6 -Xaa 7 It may have the amino acid sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 22 or 23. In some embodiments, Xaa 6 is L-glutamic acid or D-glutamic acid. In such embodiments, Xaa 3 -Xaa 4 -Xaa 5 -Xaa 6 -Xaa 7 Can have the amino acid sequence of SEQ ID NO: 21. In some embodiments, Xaa 7 In such embodiments, Xaa 3 -Xaa 4 -Xaa 5 -Xaa 6 -Xaa 7 It may have the amino acid sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 16, 17, 18, 19, 20 or 21. In some embodiments, Xaa 7 In such embodiments, Xaa 3 -Xaa 4 -Xaa 5 -Xaa 6 -Xaa 7 It may have the amino acid sequence of SEQ ID NO: 12, 13, 14, 15 or 23. In some embodiments, Xaa 8 Absent. In some further embodiments, A is acetyl terminated or polyethylene glycol. In some further embodiments, B is an amide terminated amino acid residue.

[0013] In a second aspect, the present disclosure provides a peptide compound as described in Table 3.

[0014] In a third aspect, the present disclosure provides a combination of a first peptide compound as defined herein and a second peptide compound as defined herein.

[0015] In a fourth aspect, the present disclosure provides a peptide compound of Formula II:

[0016] A-Xaa 1 -Xaa A -Xaa B -Xaa 2 -Xaa 3 -Xaa 4 -Xaa 5 -Xaa 6 -Xaa 7 -Xaa C -Xaa D -Xaa E -Xaa 8 -B(II)

[0017] wherein: A is present or absent and is a moiety that improves the circulating half-life of the peptide compound; Xaa 1 Exist or not, when present, Xaa 1 is one or more L-amino acid residues or D-amino acid residues; Xaa A Exist or not, when present, Xaa A is L-serine or D-serine; Xaa B Exist or not, when present, Xaa B is L-serine or D-serine; Xaa 2 Exist or not, when present, Xaa 2 is L-serine or D-serine; Xaa 3 is L-lysine, D-lysine, L-arginine or D-arginine; Xaa 4 is L-valine, D-valine, L-isoleucine, D-isoleucine, L-leucine, D-leucine, D-glycine, L-glycine, D-alanine or L-alanine; Xaa 5 is L-serine, D-serine, L-threonine or D-threonine; Xaa 6 is L-glutamine, D-glutamine, L-glutamic acid or D-glutamic acid; Xaa 7 Exist or not, when present, Xaa 7 is L-serine or D-serine; Xaa C Exist or not, when present, Xaa Cis L-asparagine or D-asparagine; Xaa D Exist or not, when present, Xaa D Is L-phenylalanine or D-phenylalanine; Xaa E Exist or not, when present, Xaa E is L-alanine or D-alanine; Xaa 8 Exist or not, when present, Xaa 8 is one or more L-amino acid residues or D-amino acid residues; and B is present or absent and is a moiety that improves the circulation half-life of the peptide compound.

[0018] In a fourth aspect, the present disclosure provides a pharmaceutical composition comprising (i) a peptide compound as defined herein or a combination as defined herein, and (ii) a pharmaceutically acceptable carrier. In one embodiment, the pharmaceutical composition is formulated for oral administration or parenteral administration. In another embodiment, the first peptide compound of the combination is formulated for administration together with the second peptide compound of the combination.

[0019] In a fifth aspect, the present disclosure provides a method for promoting endogenous steroid production in a cell, the method comprising contacting the cell with at least one of the following: a peptide compound defined herein, a combination defined herein, or a pharmaceutical composition defined herein, to promote endogenous steroid production in the cell. The present disclosure also provides a peptide compound defined herein, a combination defined herein, or a pharmaceutical composition defined herein for promoting endogenous steroid production in a cell. The present invention also provides a peptide compound defined herein, or a combination defined herein, for use in the preparation of a drug for promoting endogenous steroid production in a cell. The present disclosure also provides a peptide compound defined herein, a combination defined herein, or a pharmaceutical composition defined herein, for promoting endogenous steroid production in a cell. In one embodiment, the steroid is testosterone. In another embodiment, the cell is in vivo. In another embodiment, the method further comprises administering a therapeutically effective amount of a peptide compound, a combination, or a pharmaceutical composition to a subject in need thereof and including the cell. In one embodiment, the subject is a mammal, such as, for example, a male. In another embodiment, the cell is from the testis, such as, for example, a testicular interstitial cell. In one embodiment, the method, use, peptide compound, combination or pharmaceutical composition is used to prevent, treat and / or alleviate the symptoms of a condition associated with hypogonadism. In one embodiment, the condition associated with hypogonadism is at least one of infertility, aging, decreased libido, sexual dysfunction, mood changes, fatigue, decreased lean body mass, decreased bone mineral density, increased visceral fat, wasting or metabolic syndrome. In another embodiment, hypogonadism is primary hypogonadism, secondary hypogonadism, tertiary hypogonadism or acquired hypogonadism.

[0020] Throughout this application, different terms are used and some of them are more precisely defined herein.

[0021] 14-3-3ε protein. As used in the context of the present disclosure, the 14-3-3ε protein is encoded by the YWHAE gene and is an adaptor protein involved in regulating a wide range of general and specific signal transduction pathways. The 14-3-3ε protein is usually bound to a large number of partners by recognizing phosphoserine or phosphothreonine motifs. It has been shown that the 14-3-3ε protein interacts with the VDAC1 protein, and this interaction regulates (e.g., reduces) endogenous steroid production, such as, endogenous testosterone protein.

[0022] VDAC1 protein. As used in the context of the present disclosure, VDAC1 protein, also known as voltage-dependent anion selective channel protein 1, is encoded by the VDAC1 gene and forms a channel through the mitochondrial outer membrane and plasma membrane. As shown herein, VDAC1 protein can interact with TSPO protein to form a mitochondrial channel for cholesterol transport. VDAC1 has been recorded in humans (Accession No. NP_003365), mice (Accession No. NP_035824) and rats (Accession No. NP_112643). BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Having thus generally described the nature of the present invention, reference will now be made to the accompanying drawings showing by way of illustration preferred embodiments of the present invention and in which:

[0024] Figures 1A to 1C Subcutaneous infusion of a VDAC1-derived tetrapeptide was shown to increase plasma testosterone levels. Figure 1A ) Testosterone levels in 54-day-old Brown-Norway rats implanted with osmotic pumps delivering water (○) or 377 ng / kg / d of N163-166 peptide (●, RVTQ, SEQ ID NO: 2); N=12; results are shown as mean±SD; ***p<0.001. ( Figure 1B ) Protein sequence alignment, identifying amino acid variations (underlined) of RVTQ in other species. Figure 1C ) Testosterone levels in 54- and 65-day-old Brown-Norway rats infused with water (○), RVSQ (●, SEQ ID NO: 3), RITQ (▲, SEQ ID NO: 4), or KITQ (■, SEQ ID NO: 5). N=6; Results are shown as mean±SD; ****p<0.0001, *p<0.05.

[0025] Figure 2A and Figure 2B Modification of the RVTQ (SEQ ID NO: 2) core was shown to increase plasma testosterone levels after oral administration. Figure 2A ) in water (○), RVTQ (●, SEQ ID NO: 2) or RVTQdS-NH 2 (■, SEQ ID NO:6) Testosterone levels in Brown-Norway rats 3 hours after gavage. ( Figure 2B ) Summary data are shown for mice fed with water (○), RVTQ (●, SEQ ID NO: 2) or RVTQdS-NH 2(■, SEQ ID NO: 6) Time course of testosterone levels obtained in the last two rounds (1&3hr and 2&5hr). The age of Brown-Norway rats during the experiment was 60-123 days old; N=7; results are shown as mean±SD; *p<0.05.

[0026] FIG. 3A to FIG. 3I It was shown that amino acid changes in the RVTQ (SEQ ID NO: 2) core with or without modifications increased plasma testosterone levels after oral administration. Figure 3A -H) Testosterone levels 2 hours after gavage with various modified and naked protein nuclei. ( Fig. 3I ) Normalized testosterone levels for all peptides in the order tested. Age of Brown-Norway rats during the experiment was 60-136 days old; N=8; Results are shown as mean±SD; *p<0.05; *p<0.01.

[0027] FIG. 4A to FIG. 4D A biphasic response was shown in some peptides. Two hours after gavage with different concentrations of RdVTQ (SEQ ID NO: 7), Brown-Norway rats ( Figure 4A ) testosterone and ( Figure 4B ) corticosterone levels. Two hours after gavage with different concentrations of RITQ (SEQ ID NO: 4), Brown-Norway rats ( Figure 4C ) testosterone and ( Figure 4D ) Corticosterone levels. The age of Brown-Norway rats during the experiment was 88-138 days old; N=8; results are shown as mean±SD; *p<0.05.

[0028] Figure 5 Confirmation of oral peptides with stable spectra is shown. Heat map depicts the average percentage of plasma testosterone levels in rats 2 hours after treatment with various peptides and concentrations relative to control levels. Rats treated with approximately 10 μg peptide / kg and 220 μg peptide / kg, N=7, and for 420 μg peptide / kg, N=6. The age of the Brown-Norway rats during the experiment was 82-131 days old; *p<0.05.

[0029] FIG. 6A to FIG. 6G A combination of very low doses of the two peptides administered orally was shown to increase plasma testosterone levels. 6A to 6F ) Testosterone levels in 91-134 day old Brown-Norway rats treated with a 2-peptide combination at approximately 10 μg peptide / kg. ( Figure 6G ) Normalized testosterone levels of all peptides in the order tested. N=8; Results are shown as mean±SD; *p<0.05; *p<0.01.

[0030] Figure 7 Modifications to the peptide core were shown to increase plasma stability. Results are shown as % of peptide levels relative to the first time point (1 min). N=3; results are shown as mean±SEM.

[0031] FIG. 8A to FIG. 8C Pharmacokinetics of selected peptides following a single oral dose are shown. Fig. 8A -C) Circulating levels after a single oral administration of various peptides. The age of Brown-Norway rats during the experiments was 71-110 days old; N=4 in two rounds; results are shown as mean±SEM.

[0032] 9A to 9E Modifications to the RVTQ (SEQ ID NO: 2) core were shown not to increase plasma testosterone levels after oral administration. Results are shown as plasma testosterone levels in Brown-Norway rats 3 hours after gavage with water, RVTQ (SEQ ID NO: 2) or various core modifications.

[0033] Fig.10 The doses used during the oral screening experiments are shown for each peptide tested.

[0034] FIG. 11A to FIG. 11J It was shown that some modifications to the RVTQ (SEQ ID NO: 2) core sequence did not increase plasma testosterone levels. Results are shown as plasma testosterone levels in Brown-Norway rats 3 hours after gavage with water or various peptides.

[0035] FIG. 12A to FIG. 12I Some of the dual peptide combinations shown to increase testosterone levels as monotherapy at higher levels did not increase plasma testosterone levels at about 10 μg / kg. Results are shown as plasma testosterone levels in Brown-Norway rats 3 hours after gavage with water or different combinations of peptides.

[0036] FIG. 13A to FIG. 13J Various deletions of TV159-172 were shown to retain androgenic activity. FIG. 13A to FIG. 13I ) Results are shown as plasma testosterone levels in Brown-Norway rats implanted subcutaneously with osmotic pumps delivering increasing concentrations of various TV159-172 derivatives for one week. The peptide consists of a TAT sequence, a glycine linker, and a VDAC1-derived sequence shown in bold. ( Fig.13J ) Pooled testosterone levels for all water-treated rats and each peptide, independent of dose. N=6; results are shown as mean±SD.

[0037] FIG. 14A to FIG. 14I It was shown that various deletions of TV159-172 could still increase corticosterone levels. FIG. 14A to FIG. 14I ) Results are shown as plasma corticosterone levels in Brown-Norway rats implanted subcutaneously with osmotic pumps delivering increasing concentrations of various TV159-172 derivatives for one week. The peptides consist of a TAT sequence, a glycine linker, and a VDAC1-derived sequence shown in bold. N=6; results are shown as mean ± standard deviation.

[0038] FIG. 15A to FIG. 15C Subcutaneous administration of the tetrapeptide RVTQ was shown to increase testosterone levels. Fig.15A ) Summary of peptides used in the deletion experiments delivered subcutaneously. Results are shown for testosterone in Brown-Norway rats implanted subcutaneously with osmotic pumps delivering increasing concentrations of RVTQ peptide for one week. Fig. 15B ) and corticosterone ( Fig. 15C ) Plasma levels. N = 6; results are shown as mean ± SD. DETAILED DESCRIPTION

[0039] Cholesterol is the precursor of all steroids, and the rate at which cholesterol enters mitochondria directly affects steroid biosynthesis. Voltage-sensitive anion-selective protein 1 (VDAC1) is located in the outer mitochondrial membrane and forms a permeable channel that allows molecules to cross mitochondria. In steroidogenic tissues, VDAC1 is part of a protein complex that regulates cholesterol entry into mitochondria. A fusion peptide carrying a TAT cell-penetrating tag, a glycine linker, and a 14-amino acid VDAC1 sequence is known to increase testosterone levels in vitro and in vivo (Aghazadeh et al., 2014).

[0040] The present disclosure relates to peptide compounds and combinations of peptide compounds, which can promote steroid production when administered orally, and testosterone production in some embodiments. In some embodiments, peptide compounds and / or their related combinations can not change adrenal steroid production, because they can not significantly increase corticosterone levels. In some other embodiments, peptide compounds and / or their related combinations increase the levels of both testosterone and corticosterone. In some other embodiments, peptide compounds and / or their related combinations escape HPG suppression, because they can increase steroid levels during the 2 / 3 hour window. Therefore, peptide compounds and / or related combinations can be easily used for multiple administrations.

[0041] Peptide compounds

[0042] The peptide compounds of the present disclosure include an amino acid core that can be changed at the amino terminus and / or carboxyl terminus to improve the circulation half-life. The amino acid core of the peptide compounds of the present disclosure can be composed of L-amino acid residues, D-amino acid residues and combinations thereof. In some embodiments, the amino acid core of the peptide compounds of the present disclosure can be composed partially or only of peptide mimetics corresponding to the amino acid residues disclosed herein.

[0043] The peptide compounds of the present disclosure may have the following formula (I):

[0044] A-Xaa 1 -Xaa 2 -Xaa 3 -Xaa 4 -Xaa 5 -Xaa 6 -Xaa 7 -Xaa 8 -B(I)

[0045] In the peptide compound of formula I, residues A, Xaa 1 , Xaa 2 , Xaa 7 , Xaa 8 and B is optional, and residue Xaa 3 , Xaa 4 , Xaa 5 and Xaa 6 As used in the context of this disclosure, "amino acid core" refers to "Xaa 1 -Xaa 2 -Xaa 3 -Xaa 4 -Xaa 5 -Xaa 6 -Xaa 7 -Xaa 8 " and excludes A and B residues. The symbol "-" in the amino acid core refers to the binding peptide formed between two consecutive amino acid residues. In addition, the "amino acid core" always includes "Xaa 3 -Xaa 4 -Xaa 5 -Xaa 6 ”.

[0046] As indicated above, at least the amino acid core of the peptide compound comprises "Xaa 3 -Xaa 4 -Xaa 5 -Xaa 6 ".Residue Xaa 3Xaa may be any amino acid residue, and is preferably lysine or arginine (each independently selected as a D-enantiomer or an L-enantiomer). 3 is arginine (D-enantiomer or L-enantiomer). Residue Xaa 4 It can be any amino acid residue, and is preferably valine, isoleucine, leucine, glycine or alanine (each independently selected as a D-enantiomer or an L-enantiomer) or a non-natural amino acid similar in structure to valine. 4 is valine (D-enantiomer or L-enantiomer). Residue Xaa 5 Xaa may be any amino acid residue, and is preferably serine or threonine (each independently selected as a D-enantiomer or an L-enantiomer). 5 is serine (D-enantiomer or L-enantiomer). Residue Xaa 6 Xaa may be any amino acid residue, and is preferably glutamine or glutamic acid (each independently selected as a D-enantiomer or an L-enantiomer). 6 is glutamine (D-enantiomer or L-enantiomer). In some embodiments, in the peptide compounds of the present disclosure, Xaa 5 is serine and Xaa 6 is glutamine (both independently selected from the D-enantiomer or the L-enantiomer).

[0047] In some embodiments, the peptide compounds of the present disclosure may have the following amino acid core consensus sequence, and optionally include additional amino acid residues or modifications:

[0048] XA 3 -Xaa 4 -Xaa 5 -Xaa 6 (Formula Ia, SEQ ID NO: 24)

[0049] XA 2 -Xaa 3 -Xaa 4 -Xaa 5 -Xaa 6 -Xaa 7 (Formula Ib, SEQ ID NO: 25) or

[0050] XA 1 -Xaa 2 -Xaa 3 -Xaa 4 -Xaa 5-Xaa 6 -Xaa 7 -Xaa 8 (Formula Ic, SEQ ID NO:26).

[0051] In Formulas Ia, Ib, and Ic, Xaa 3 , Xaa 4 , Xaa 5 and Xaa 6 As defined above. Residue Xaa 1 Can be present or absent, when present, Xaa 1 It can be any amino acid residue. 2 Can be present or absent, Xaa 2 It can be any amino acid residue, and is preferably serine (D-enantiomer or L-enantiomer). Residue Xaa 7 Can be present or absent, when present, Xaa 7 It can be any amino acid residue, and is preferably serine (D-enantiomer or L-enantiomer). Residue Xaa 8 Can be present or absent, when present, Xaa 8 It can be any amino acid residue.

[0052] Peptide compounds having an amino acid core of Formula Ib include those having one of the following amino acid cores (all derived from SEQ ID NO: 25, √ indicates the presence of an amino acid residue, and × indicates the absence of an amino acid residue):

[0053] <![CDATA[Xaa 2 ]]> <![CDATA[Xaa 3 ]]> <![CDATA[Xaa 4 ]]> <![CDATA[Xaa 5 ]]> <![CDATA[Xaa 6 ]]> <![CDATA[Xaa 7 ]]> √ √ √ √ √ √ × √ √ √ √ √ √ √ √ √ √ × × √ √ √ √ ×

[0054] Peptide compounds having an amino acid core of Formula Ic include those having one of the following amino acid cores (all derived from SEQ ID NO: 26, √ indicates the presence of an amino acid residue, and × indicates the absence of an amino acid residue):

[0055] <![CDATA[Xaa 1 ]]> <![CDATA[Xaa 2 ]]> <![CDATA[Xaa 3 ]]> <![CDATA[Xaa 4 ]]> <![CDATA[Xaa 5 ]]> <![CDATA[Xaa 6 ]]> <![CDATA[Xaa 7 ]]> <![CDATA[Xaa 8 ]]> √ √ √ √ √ √ √ √ × √ √ √ √ √ √ √ √ × √ √ √ √ √ √ √ √ √ √ √ √ × √ √ √ √ √ √ √ √ × × × √ √ √ √ √ √ × √ √ √ √ √ × √ × √ √ √ √ √ √ × × × √ √ √ √ × √ × × √ √ √ √ √ × × × √ √ √ √ × × <![CDATA[Xaa 1 ]]> <![CDATA[Xaa 2 ]]> <![CDATA[Xaa 3 ]]> <![CDATA[Xaa 4 ]]> <![CDATA[Xaa 5 ]]> <![CDATA[Xaa 6 ]]> <![CDATA[Xaa 7 ]]> <![CDATA[Xaa 8 ]]> × √ √ √ √ √ × × √ × √ √ √ √ × √ √ × √ √ √ √ √ × √ × √ √ √ √ × × √ √ √ √ √ √ × ×

[0056] Table 1 summarizes the amino acid cores of the peptide compounds of the Examples. In some embodiments, the peptide compounds of the present disclosure have an amino acid core as described in Table 1 and optionally include additional amino acid residues or modifications (at the amino or carboxyl terminus). In some embodiments, the peptide compounds of the present disclosure have an amino acid core as described in Table 1 and have no additional amino acid residues or modifications (at the amino or carboxyl terminus).

[0057] Table 1. Core amino acid sequences of peptide compounds of the examples. The names in the first row of the table refer to the residues of formula (I). The symbol "--" means that the amino acid residue is not present in the core amino acid sequence. The symbol "*" means that the amino acid residue can be either the L-enantiomer or the D-enantiomer. Compound # refers to the compound name presented in Table 3.

[0058]

[0059] The peptide compound of the present disclosure or the amino acid core of the peptide compound can only include L-amino acid residues. In such embodiments, the peptide compound and the amino acid core can be produced in a host (such as, cell or cell line), and the host has been genetically engineered to express the peptide compound or the amino acid core. The amino acid core or the peptide compound produced in the recombinant host can be substantially purified or further chemically modified before being used to regulate steroid production.

[0060] The peptide compounds of the present disclosure may have the following formula (II):

[0061] A-Xaa 1 -Xaa A -Xaa B -Xaa 2 -Xaa 3 -Xaa 4 -Xaa 5 -Xaa 6 -Xaa 7 -Xaa C -Xaa D -Xaa E -Xaa 8 -B(II)

[0062] In the peptide compound of formula II, residues A, Xaa 1 , Xaa A , Xaa B、 XA 2 , Xaa 7 , Xaa C , Xaa D , Xaa E , Xaa 8 and B is optional, and residue Xaa 3 , Xaa 4 , Xaa 5 and Xaa 6 As used in the context of the present disclosure, in the context of a peptide having formula (II), "amino acid core" refers to "Xaa 1 -Xaa A -Xaa B -Xaa 2 -Xaa3 -Xaa 4 -Xaa 5 -Xaa 6 -Xaa 7 -Xaa C -Xaa D -Xaa E -Xaa 8 " and excludes A residues and B residues. The symbol "-" in the amino acid core refers to the binding peptide formed between two consecutive amino acid residues. In addition, the "amino acid core" always includes "Xaa 3 -Xaa 4 -Xaa 5 -Xaa 6 ”.

[0063] As indicated above, at least the amino acid core of the peptide compound comprises "Xaa 3 -Xaa 4 -Xaa 5 -Xaa 6 ".Residue Xaa 3 Xaa may be any amino acid residue, and is preferably lysine or arginine (each independently selected as a D-enantiomer or an L-enantiomer). 3 is arginine (D-enantiomer or L-enantiomer). Residue Xaa 4 Xaa may be any amino acid residue, and is preferably valine, isoleucine or leucine (each independently selected as a D-enantiomer or an L-enantiomer). 4 is valine (D-enantiomer or L-enantiomer). Residue Xaa 5 Xaa may be any amino acid residue, and is preferably serine or threonine (each independently selected as a D-enantiomer or an L-enantiomer). 5 is serine (D-enantiomer or L-enantiomer). Residue Xaa 6 Xaa may be any amino acid residue, and is preferably glutamine or glutamic acid (each independently selected as a D-enantiomer or an L-enantiomer). 6 is glutamine (D-enantiomer or L-enantiomer). In some embodiments, in the peptide compounds of the present disclosure, Xaa 5 is serine and Xaa 6 is glutamine (both independently selected from the D-enantiomer or the L-enantiomer).

[0064] In some embodiments, the peptide compounds of the present disclosure may have the following amino acid core consensus sequence, and optionally include additional amino acid residues or modifications:

[0065] XA 1 -Xaa A -Xaa B -Xaa 2 -Xaa 3 -Xaa 4 -Xaa 5 -Xaa 6 -Xaa 7 -Xaa 8 (Formula IIa, SEQ ID NO:27);

[0066] XA 1 -Xaa 2 -Xaa 3 -Xaa 4 -Xaa 5 -Xaa 6 -Xaa 7 -Xaa C -Xaa D -Xaa E -Xaa 8 (Formula IIb, SEQ ID NO: 28);

[0067] XA 1 -Xaa A -Xaa B -Xaa 2 -Xaa 3 -Xaa 4 -Xaa 5 -Xaa 6 -Xaa 7 -Xaa C -Xaa D -Xaa E -Xaa 8 (Formula IIc, SEQ ID NO: 29).

[0068] In Formula IIa, IIb, and IIc, Xaa 3 , Xaa 4 , Xaa 5 and Xaa 6 As defined above. Residue Xaa 1 Can be present or absent, when present, Xaa 1 Can be any amino acid residue. Residue Xaa A Can be present or absent, when present, Xaa AIt can be any amino acid residue, preferably serine (D-enantiomer or L-enantiomer). Residue Xaa B Can be present or absent, when present, Xaa B It can be any amino acid residue, preferably lysine (D-enantiomer or L-enantiomer). Residue Xaa 2 Can be present or absent, Xaa 2 It can be any amino acid residue, and is preferably serine (D-enantiomer or L-enantiomer). Residue Xaa 7 Can be present or absent, when present, Xaa 7 It can be any amino acid residue, and is preferably serine (D-enantiomer or L-enantiomer). Residue Xaa C Can be present or absent, when present, Xaa C It can be any amino acid residue, preferably asparagine (D-enantiomer or L-enantiomer). Residue Xaa D Can be present or absent, when present, Xaa D It can be any amino acid residue, preferably phenylalanine (D-enantiomer or L-enantiomer). Residue Xaa E Can be present or absent, when present, Xaa E It can be any amino acid residue, preferably alanine (D-enantiomer or L-enantiomer). Residue Xaa 8 Can be present or absent, when present, Xaa 8 It can be any amino acid residue.

[0069] Peptide compounds having an amino acid core of Formula IIa include those having one of the following amino acid cores (all derived from SEQ ID NO: 27, √ indicates the presence of an amino acid residue, and × indicates the absence of an amino acid residue):

[0070] <![CDATA[Xaa 1 ]]> <![CDATA[Xaa A ]]> <![CDATA[Xaa B ]]> <![CDATA[Xaa 2 ]]> <![CDATA[Xaa 3 ]]> <![CDATA[Xaa 4 ]]> <![CDATA[Xaa 5 ]]> <![CDATA[Xaa 6 ]]> <![CDATA[Xaa 7 ]]> <![CDATA[Xaa 8 ]]> √ √ √ √ √ √ √ √ √ √ √ √ √ √ √ √ √ √ √ × √ √ √ √ √ √ √ √ × √ √ √ √ √ √ √ √ √ × × × √ √ √ √ √ √ √ √ √ × √ √ √ √ √ √ √ √ × × √ √ √ √ √ √ √ × √ × √ √ √ √ √ √ √ × × × × √ √ √ √ √ √ √ √ × × √ √ √ √ √ √ √ × × × √ √ √ √ √ √ × √ × × √ √ √ √ √ √ × × × × × √ √ √ √ √ √ √ × × × √ √ √ √ √ √ × × × × √ √ √ √ √ × √ × × × √ √ √ √ √ × × × × × × √ √ √ √ √ √ × × × × √ √ √ √ √ × × × × × √ √ √ √ × √ × × × × √ √ √ √ × ×

[0071] Peptide compounds having an amino acid core of Formula IIb include those having one of the following amino acid cores (all derived from SEQ ID NO: 28, √ indicates the presence of an amino acid residue, and × indicates the absence of an amino acid residue):

[0072]

[0073]

[0074] Peptide compounds having an amino acid core of Formula IIc include those having one of the following amino acid cores (all derived from SEQ ID NO: 29, √ indicates the presence of an amino acid residue, and × indicates the absence of an amino acid residue):

[0075]

[0076]

[0077]

[0078] Table 2 summarizes the amino acid core of the peptide compounds of the examples. In some embodiments, the peptide compounds of the present disclosure have an amino acid core as described in Table 2 and optionally include additional amino acid residues or modifications (at the amino or carboxyl terminus). In some embodiments, the peptide compounds of the present disclosure have an amino acid core as described in Table 1 and have no additional amino acid residues or modifications (at the amino or carboxyl terminus).

[0079] Table 2. Core amino acid sequences of peptide compounds of the examples. The names in the first row of the table refer to the residues of formula (II). The symbol "--" means that the amino acid residue is not present in the core amino acid sequence. The symbol "*" means that the amino acid residue can be either the L-enantiomer or the D-enantiomer. Compound # refers to the compound name presented in Table 3.

[0080] <![CDATA[Xaa A ]]> <![CDATA[Xaa B ]]> <![CDATA[Xaa 2 ]]> <![CDATA[Xaa 3 ]]> <![CDATA[Xaa 4 ]]> <![CDATA[Xaa 5 ]]> <![CDATA[Xaa 6 ]]> <![CDATA[Xaa 7 ]]> <![CDATA[Xaa C ]]> <![CDATA[Xaa D ]]> <![CDATA[Xaa E ]]> SEQ ID NO: R V T Q 2 S R V T Q 17 R V T Q S 12 K S R V T Q S 30 S R V T Q S N F 31 K S R V T Q S N F A 32 S K S R V T Q S N F 33 S K S R V T Q S N F A 34

[0081] The peptide compound of the present disclosure or the amino acid core of the peptide compound can only include L-amino acid residues. In such embodiments, the peptide compound and the amino acid core can be produced in a host (such as, cell or cell line), and the host has been genetically engineered to express the peptide compound or the amino acid core. The amino acid core or the peptide compound produced in the recombinant host can be substantially purified or further chemically modified before being used to regulate steroid production.

[0082] Alternatively, the peptide compounds of the present disclosure or the amino acid core of the peptide compounds may include at least one and in some embodiments only D-amino acid residues. In such embodiments, the peptide compounds and amino acid cores may be chemically synthesized and substantially purified before being used to modulate steroid production or further chemically modified.

[0083] In some embodiments, the amino acid core can be used as a peptide compound without further modification (change, motification). Alternatively, the amino acid core can be modified at its amino terminal and / or carboxyl terminal to further improve the therapeutic profile of the peptide compound. In the peptide compound of Formula I, these other modifications are labeled as residues "A" and "B". Modifications "A" and "B" can be any modifications performed on the amino acid core, and in some embodiments, these modifications will retain the ability of the peptide compound to promote endogenous testosterone synthesis and increase the circulation half-life of the peptide compound (when compared with the corresponding compound lacking modification). Residues A and B are independently selected from amide end-capping, acetyl end-capping, non-charged, hydrophilic and / or non-toxic polymers (such as polyethylene glycol polymers). In the embodiment where it is desired that the peptide compound is positioned in a cell or an organism, residues A and B can be marks, such as biotin or fluorescent labels or isotope (radioactive or non-radioactive) labels.

[0084] In some embodiments, the peptide compounds of the present disclosure include residue A and may have the following formula:

[0085] A-Xaa 1 -Xaa 2 -Xaa 3 -Xaa 4 -Xaa 5 -Xaa 6 -Xaa 7 -Xaa 8 -B(I)

[0086] A-Xaa 1 -Xaa 2 -Xaa 3 -Xaa 4 -Xaa 5 -Xaa 6 -Xaa 7 -Xaa 8 (Id)

[0087] In such embodiments, residues A, Xaa 1 (when present), Xaa 2 (when present), Xaa 3 , Xaa 4 , Xaa 5 , Xaa 6 , Xaa 7 (when present), Xaa 8 A (when present) and B (when present) are as defined above.

[0088] In some embodiments, the peptide compounds of the present disclosure do not include residue A and may have the following formula:

[0089] XA 1 -Xaa 2 -Xaa 3 -Xaa 4 -Xaa 5 -Xaa 6 -Xaa 7 -Xaa 8 -B(Ie)

[0090] XA 1 -Xaa 2 -Xaa 3 -Xaa 4 -Xaa 5 -Xaa 6 -Xaa 7 -Xaa 8 (If)

[0091] In such embodiments, residue Xaa 1 (when present), Xaa 2 (when present), Xaa 3 , Xaa 4 , Xaa 5 , Xaa 6 , Xaa 7 (when present), Xaa 8 A (when present) and B (when present) are as defined above.

[0092] In some embodiments, the peptide compounds of the present disclosure include residue B and may have the following formula:

[0093] A-Xaa 1 -Xaa 2 -Xaa 3 -Xaa 4 -Xaa 5 -Xaa 6 -Xaa 7 -Xaa 8 -B(I)

[0094] XA 1 -Xaa 2 -Xaa 3 -Xaa 4 -Xaa 5 -Xaa 6 -Xaa 7 -Xaa 8 -B(Ie)

[0095] In such embodiments, residues A (when present), Xaa 1(when present), Xaa 2 (when present), Xaa 3 , Xaa 4 , Xaa 5 , Xaa 6 , Xaa 7 (when present), Xaa 8 (when present) and B are as defined above.

[0096] In some embodiments, the peptide compounds of the present disclosure do not include residue B and may have the following formula:

[0097] A-Xaa 1 -Xaa 2 -Xaa 3 -Xaa 4 -Xaa 5 -Xaa 6 -Xaa 7 -Xaa 8 (Id)

[0098] XA 1 -Xaa 2 -Xaa 3 -Xaa 4 -Xaa 5 -Xaa 6 -Xaa 7 -Xaa 8 (If)

[0099] In such embodiments, residues A (when present), Xaa 1 (when present), Xaa 2 (when present), Xaa 3 , Xaa 4 , Xaa 5 , Xaa 6 , Xaa 7 (when present) and Xaa 8 (when present) is as defined above.

[0100] In some embodiments, the peptide compounds of the present disclosure include residue A and may have the following formula:

[0101] A-Xaa 1 -Xaa A -Xaa B -Xaa 2 -Xaa 3 -Xaa 4 -Xaa 5 -Xaa 6 -Xaa 7 -XaaC -Xaa D -Xaa E -Xaa 8 -B(II)

[0102] A-Xaa 1 -Xaa A -Xaa B -Xaa 2 -Xaa 3 -Xaa 4 -Xaa 5 -Xaa 6 -Xaa 7 -Xaa C -Xaa D -Xaa E -Xaa 8 (IId)

[0103] In such embodiments, residues A, Xaa 1 (when present), Xaa A (when present), Xaa B (when present), Xaa 2 (when present), Xaa 3 , Xaa 4 , Xaa 5 , Xaa 6 , Xaa 7 (when present), Xaa C (when present), Xaa D (when present), Xaa E (when present), Xaa 8 A (when present) and B (when present) are as defined above.

[0104] In some embodiments, the peptide compounds of the present disclosure do not include residue A and may have the following formula:

[0105] XA 1 -Xaa A -Xaa B -Xaa 2 -Xaa 3 -Xaa 4 -Xaa 5 -Xaa 6 -Xaa 7 -Xaa C -Xaa D -Xaa E -Xaa 8 -B(IIe)

[0106] XA1 -Xaa A -Xaa B -Xaa 2 -Xaa 3 -Xaa 4 -Xaa 5 -Xaa 6 -Xaa 7 -Xaa C -Xaa D -Xaa E -Xaa 8 (IIf)

[0107] In such embodiments, residue Xaa 1 (when present), Xaa A (when present), Xaa B (when present), Xaa 2 (when present), Xaa 3 , Xaa 4 , Xaa 5 , Xaa 6 , Xaa 7 (when present), Xaa C (when present), Xaa D (when present), Xaa E (when present), Xaa 8 A (when present) and B (when present) are as defined above.

[0108] In some embodiments, the peptide compounds of the present disclosure include residue B and may have the following formula:

[0109] A-Xaa 1 -Xaa A -Xaa B -Xaa 2 -Xaa 3 -Xaa 4 -Xaa 5 -Xaa 6 -Xaa 7 -Xaa C -Xaa D -Xaa E -Xaa 8 -B(II)

[0110] XA 1 -Xaa A -Xaa B -Xaa 2 -Xaa 3 -Xaa 4 -Xaa5 -Xaa 6 -Xaa 7 -Xaa C -Xaa D -Xaa E -Xaa 8 -B(IIe)

[0111] In such embodiments, residues A (when present), Xaa 1 (when present), Xaa A (when present), Xaa B (when present), Xaa 2 (when present), Xaa 3 , Xaa 4 , Xaa 5 , Xaa 6 , Xaa 7 (when present), Xaa C (when present), Xaa D (when present), Xaa E (when present), Xaa 8 A (when present) and B (when present) are as defined above.

[0112] In some embodiments, the peptide compounds of the present disclosure do not include residue B and may have the following formula:

[0113] A-Xaa 1 -Xaa A -Xaa B -Xaa 2 -Xaa 3 -Xaa 4 -Xaa 5 -Xaa 6 -Xaa 7 -Xaa C -Xaa D -Xaa E -Xaa 8 (IId)

[0114] XA 1 -Xaa A -Xaa B -Xaa 2 -Xaa 3 -Xaa 4 -Xaa 5 -Xaa 6 -Xaa 7 -Xaa C -Xaa D -Xaa E-Xaa 8 (IIf)

[0115] In such embodiments, residues A (when present), Xaa 1 (when present), Xaa A (when present), Xaa B (when present), Xaa 2 (when present), Xaa 3 , Xaa 4 , Xaa 5 , Xaa 6 , Xaa 7 (when present), Xaa C (when present), Xaa D (when present), Xaa E (when present) and Xaa 8 (when present) is as defined above.

[0116] Combinations of peptide compounds

[0117] The present disclosure provides a combination of peptide compounds for increasing endogenous steroid production (including endogenous testosterone production). The combination comprises at least two or more different peptide compounds. In the combination of the present disclosure, the peptide compounds are considered to be different because they are structurally different from each other. If the peptide compounds of the combination are different in at least one amino acid identity in its amino acid core, the length of the continuous amino acid residues in its amino acid core, the amino acid enantiomer for amino, the existence, absence and identity of the modification carried out by its amino terminal, the existence, absence and identity of the modification carried out by its carboxyl terminal, then they are considered to be "different". In some embodiments, the combination comprises no more than two peptide compounds.

[0118] To achieve the desired therapeutic endpoint (eg, increase in endogenous steroid production), the combined peptide compounds may be provided in the same dosage form or in discrete dosage forms.The peptide compounds of the present disclosure may be (or are intended to be) co-administered or sequentially administered.

[0119] The combinations of the present disclosure encompass combinations of at least two different peptide compounds of formula (I), (Ia), (Ib), (Ic), (Id), (Ie), (If), (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf) described in Tables 1, 2 or 3. In some specific embodiments, the combinations of the present disclosure relate to a combination comprising (or consisting essentially of) at least two different peptide compounds as described in Table 3. For example, the combination comprises (or consists essentially of) the following pairs of peptide compounds described in Table 3: 1 and 2, 1 and 3, 1 and 4, 1 and 5, 1 and 6, 1 and 7, 1 and 8, 1 and 9, 1 and 10, 1 and 11, 1 and 12, 1 and 13, 1 and 14, 1 and 15, 1 and 16, 1 and 17, 1 and 18, 1 and 19, 1 and 20, 1 and 21, 1 and 22, 1 and 23, 1 and 24, 1 and 25, 1 and 26, 1 and 27, 1 and 28, 1 and 29, 1 and 30, 1 and 31, 1 and 32, 1 and 33, 1 and 34, 1 and 35, 1 and 36, 1 and 37, 1 and 38, 1 and 39, and 39, 2 and 3, 2 and 4, 2 and 5, 2 and 6, 2 and 7, 2 and 8, 2 and 9, 2 and 10, 2 and 11, 2 and 12, 2 and 13, 2 and 14, 2 and 15, 2 and 16, 2 and 17, 2 and 18, 2 and 19, 2 and 20, 2 and 21, 2 and 22, 2 and 23, 2 and 24, 2 and 25, 2 and 26, 2 and 27, 2 and 28, 2 and 29, 2 and 30, 2 and 31, 2 and 32, 2 and 33, 2 and 34, 2 and 35, 2 and 36, 2 and 37, 2 and 38, 2 and 39, 3 and 4, 3 and 5, 3 and 6, 3 and 7, 3 and 8, 3 and 9, 3 and 10, 3 and 11, 3 and 12, 3 and 13, 3 and 14, 3 and 15, 3 and 16, 3 and 17, 3 and 18, 3 and 19, 3 and 20, 3 and 21, 3 and 22, 3 and 23, 3 and 24, 3 and 25, 3 and 26, 3 and 27, 3 and 28, 3 and 29, 3 and 30 , 3 and 31, 3 and 32, 3 and 33, 3 and 34, 3 and 35, 3 and 36, 3 and 37, 3 and 38, 3 and 39, 4 and 5, 4 and 6, 4 and 7, 4 and 8, 4 and 9, 4 and 10, 4 and 11, 4 and 12, 4 and 13, 4 and 14, 4 and 15, 4 and 16, 4 and 1 and 38, 4 and 39, 5 and 6, 5 and 7, 5 and 8, 5 and 9, 5 and 10, 5 and 11, 5 and 12, 5 and 13, 5 and 14, 5 and 15, 5 and 16, 5 and 17, 5 and 18, 5 and 19, 5 and 20, 5 and 21, 5 and 22, 5 and 23, 5 and 24, 5 and 25,5W26, 5W27, 5W28, 5W29, 5W30, 5W31, 5W32, 5W33, 5W34, 5W35, 5W36, 5W37, 5W38, 5W39, 6W7, 6W8, 6W9, 6W10, 6W11, 6W12, 6W13, 6W14, 6W15, 6W16, 6W17, 6W1 8, 6W19, 6W20, 6W21, 6W22, 6W23, 6W24, 6W25, 6W26, 6W27, 6W28, 6W29, 6W30, 6W31, 6W32, 6W33, 6W34, 6W35, 6W36, 6W37, 6W38, 6W39, 7W8, 7W9, 7W10, 7W11, 7wa12, 7wa13, 7wa14, 7wa15, 7wa16, 7wa17, 7wa18, 7wa19, 7wa20, 7wa21, 7wa22, 7wa23, 7wa24, 7wa25, 7wa26, 7wa27, 7wa28, 7wa29, 7wa30, 7wa31, 7wa32, 7wa33, 7wa34, 7wa35, 7wa36, 7W37, 7W38, 7W39, 8W9, 8W10, 8W11, 8W12, 8W13, 8W14, 8W15, 8W16, 8W17, 8W18, 8W19, 8W20, 8W21, 8W22, 8W23, 8W24, 8W25, 8W26, 8W27, 8W28, 8W29, 8W30, 8 W31, 8W32, 8W33, 8W34, 8W35, 8W36, 8W37, 8W38, 8W39, 9W10, 9W11, 9W12, 9W13, 9W14, 9W15, 9W16, 9W17, 9W18, 9W19, 9W20, 9W21, 9W22, 9W23, 9W24, 9W25, 9 Japanese 26, 9Japanese 27, 9Japanese 28, 9Japanese 29, 9Japanese 30, 9Japanese 31, 9Japanese 32, 9Japanese 33, 9Japanese 34, 9Japanese 35, 9Japanese 36, 9Japanese 37, 9Japanese 38, 9Japanese 39, 10Japanese 11, 10Japanese 12, 10Japanese 13, 10Japanese 14, 10Japanese 15, 10Japanese 16, 10Japanese 17, 10Japanese 18, 10Japanese 19, 10 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 12, 11, 12, 1 1W13, 11W14, 11W15, 11W16, 11W17, 11W18, 11W19, 11W20, 11W21, 11W22, 11W23, 11W24, 11W25, 11W26, 11W27, 11W28, 11W29, 11W30, 11W31, 11W32, 11W33,11 and 34, 11 and 35, 11 and 36, 11 and 37, 11 and 38, 11 and 39, 12 and 13, 12 and 14, 12 and 15, 12 and 16, 12 and 17, 12 and 18, 12 and 19, 12 and 20, 12 and 21, 12 and 22, 12 and 23, 12 and 24, 12 and 25, 12 and 26, 12 and 27, 12 and 28, 12 and 29, 12 and 30, 12 and 31, 12 and 32, 12 and 33, 12 and 34, 12 and 35, 12 and 36, 12 and 37, 12 and 38, 12 and 39, 13 and 14, 13 and 15, 13 and 16, 13 and 17, 13 and 18, 13 and 19, 13 and 20, 13 and 21, 13 and 22, 13 and 23, 13 and 24, 13 and 25, 13 and 26, 13 and 27, 13 and 28, 13 and 29, 13 and 30, 13 and 31, 13 and 32, 13 and 33, 13 and 34, 13 and 35, 13 and 36, 13 and 37, 13 and 38, 13 and 39, 14 and 15, 14 and 16, 14 and 17, 14 and 18, 14 and 19, 14 and 20, 14 and 21, 14 and 22, 14 and 23, 14 and 24, 14 and 25, 14 and 26, 14 and 27, 14 and 28, 14 and 29, 14 and 30, 14 and 31, 14 and 32, 14 and 33, 14 and 34, 14 and 35, 14 and 36, 14 and 37, 14 and 38, 14 and 39, 15 and 16, 15 and 17, 15 and 18, 15 and 19, 15 and 20, 15 and 21, 15 and 22, 15 and 23, 15 and 24, 15 and 25, 15 and 26, 15 and 27, 15 and 28, 15 and 29, 15 and 30, 15 and 31, 15 and 32, 15 and 33, 15 and 34, 15 and 35, 15 and 36, 15 and 37, 15 and 38, 15 and 39, 16 and 17, 16 and 18, 16 and 19, 16 and 20, 16 and 21, 16 and 22, 16 and 23, 16 and 24, 16 and 25, 16 and 26, 16 and 27, 16 and 28, 16 and 29, 16 and 30, 16 and 31, 16 and 32, 16 and 33, 16 and 34, 16 and 35, 16 and 36, 16 and 37, 16 and 38, 16 and 39, 17 and 18, 17 and 19, 17 and 20, 17 and 21, 17 and 22, 17 and 23, 17 and 24, 17 and 25, 17 and 26, 17 and 27, 17 and 28, 17 and 29, 17 and 30, 17 and 31, 17 and 32, 17 and 33, 17 and 34, 17 and 35, 17 and 36, 17 and 37, 17 and 38, 17 and 39, 18 and 19, 18 and 20, 18 and 21, 18 and 22, 18 and 23, 18 and 24, 18 and 25, 18 and 26, 18 and 27, 18 and 28, 18 and 29, 18 and 30, 18 and 31,18-wa-32, 18-wa-33, 18-wa-34, 18-wa-35, 18-wa-36, 18-wa-37, 18-wa-38, 18-wa-39, 19-wa-20, 19-wa-21, 19-wa-22, 19-wa-23, 19-wa-24, 19-wa-25, 19-wa-26, 19-wa-27, 19-wa-28, 19-wa-29, 19-wa-30, 19-wa-31, 19-wa-3 2, 19W33, 19W34, 19W35, 19W36, 19W37, 19W38, 19W39, 20W21, 20W22, 20W23, 20W24, 20W25, 20W26, 20W27, 20W28, 20W29, 20W30, 20W31, 20W32, 20W33, 20W 34, 20W35, 20W36, 20W37, 20W38, 20W39, 21W22, 21W23, 21W24, 21W25, 21W26, 21W27, 21W28, 21W29, 21W30, 21W31, 21W32, 21W33, 21W34, 21W35, 21W36, 2 1W37, 21W38, 21W39, 22W23, 22W24, 22W25, 22W26, 22W27, 22W28, 22W29, 22W30, 22W31, 22W32, 22W33, 22W34, 22W35, 22W36, 22W37, 22W38, 22W39, 23W24, 23W25, 23W26, 23W27, 23W28, 23W29, 23W30, 23W31, 23W32, 23W33, 23W34, 23W35, 23W36, 23W37, 23W28, 23W39, 24W25, 24W26, 24W27, 24W28, 24W29, 24W3 0, 24W31, 24W32, 24W33, 24W34, 24W35, 24W36, 24W37, 24W38, 24W39, 25W26, 25W27, 25W28, 25W29, 25W30, 25W31, 25W32, 25W33, 25W34, 25W35, 25W36, 25W 37, 25W38, 25W39, 26W27, 26W28, 26W29, 26W30, 26W31, 26W32, 26W33, 26W34, 26W35, 26W36, 26W37, 26W38, 26W39, 27W28, 27W29, 27W30, 27W31, 27W32, 2 7W33, 27W34, 27W35, 27W36, 27W37, 27W38, 27W39, 28W29, 28W30, 28W31, 28W32, 28W33, 28W34, 28W35, 28W36, 28W37, 28W38, 28W39, 29W30, 29W31, 29W32,29 and 33, 29 and 34, 29 and 35, 29 and 36, 29 and 37, 29 and 38, 29 and 39, 30 and 31, 30 and 32, 30 and 33, 30 and 34, 30 and 35, 30 and 36, 30 and 37, 30 and 38, 30 and 39, 31 and 32, 31 and 33, 31 and 34, 31 and 35, 31 and 36, 31 and 37, 31 and 38, 31 and 39, 32 and 33, 32 and 34, 32 and 35, 32 and 36, 32 and 37, 32 and 38, 32 and 39, 33 and 34, 33 and 35, 33 and 36, 33 and 37, 33 and 38, 33 and 39, 34 and 35, 34 and 36, 34 and 37, 34 and 38, 34 and 39, 35 and 36, 35 and 37, 35 and 38, 35 and 39, 36 and 37, 36 and 38, 36 and 39, 37 and 38, 37 and 39 or 38 and 39. ,

[0120] In one embodiment, the combination comprises or consists essentially of peptide compound 20 and peptide compound 1. In another embodiment, the combination comprises or consists essentially of peptide compound 20 and peptide compound 10. In a further embodiment, the combination comprises or consists essentially of peptide compound 20 and peptide compound 19. In yet another embodiment, the combination comprises or consists essentially of peptide compound 1 and peptide compound 11. In yet another embodiment, the combination comprises or consists essentially of peptide compound 1 and peptide compound 18. In yet another embodiment, the combination comprises or consists essentially of peptide compound 11 and peptide compound 18. In yet further embodiments, the combination comprises or consists essentially of peptide compound 20 and peptide compound 18. In yet further embodiments, the combination comprises or consists essentially of peptide compound 20 and peptide compound 3. In yet further embodiments, the combination comprises or consists essentially of peptide compound 1 and peptide compound 19. In yet further embodiments, the combination comprises or consists essentially of peptide compound 1 and peptide compound 13. In yet further embodiments, the combination comprises or consists essentially of peptide compound 10 and peptide compound 19. In yet further embodiments, the combination comprises or consists essentially of peptide compound 10 and peptide compound 3. In yet further embodiments, the combination comprises or consists essentially of Peptide Compound 19 and Peptide Compound 3. In yet further embodiments, the combination comprises or consists essentially of Peptide Compound 18 and Peptide Compound 3. In yet further embodiments, the combination comprises or consists essentially of Peptide Compound 1 and Peptide Compound 10. In yet further embodiments, the combination comprises or consists essentially of Peptide Compound 10 and Peptide Compound 18.

[0121] In the combinations of the present disclosure, the term "consisting essentially of" refers to the character of the combination in that it does not include additional peptide compounds but may include other therapeutic agents or non-pharmaceutical ingredients.

[0122] Therapeutic uses of peptide compounds and combinations

[0123] Peptide compounds and combinations described herein can be used to treat hypogonadism or disorders associated with hypogonadism. Hypogonadism is understood to be a weakening of the functional activity of gonads (e.g., testes and ovaries), resulting in weakening of sex hormones (e.g., testosterone, estradiol, biosynthetic progesterone, DHEA, anti-Müllerian hormone, activin and inhibin). Hypogonadism can be primary (e.g., associated with reduced androgen production in interstitial cells), secondary (e.g., insufficient gonadotropin-releasing hormone or luteinizing hormone signals) or tertiary (e.g., associated with reduced androgen production and insufficient gonadotropin-releasing hormone or luteinizing hormone signals). Hypogonadism can also be an acquired disorder caused by exposure to stress or trauma (e.g., surgery, exposure to toxic chemicals (such as, explosives and chemotherapy), exposure to therapeutic drugs or biological products, post-traumatic stress syndrome, etc.). Hypogonadism can be compensated by increasing luteinizing hormone levels. Hypogonadism can be late-onset hypogonadism and is relevant to central and gonadal insufficiency.In some embodiments, hypogonadism can be caused by trauma.Low androgen (for example, testosterone) level can be referred to as low androgenism, and low estrogen (for example, estradiol) can be referred to as low estrogenism.The disease relevant to hypogonadism includes but is not limited to infertility (due to spermatogenesis or ovulation defect or deficiency), aging, reduced libido, sexual dysfunction, mood changes, fatigue, lean body mass reduction, bone mineral density reduction, visceral fat increase, emaciation (metabolic, relevant to receiving chemotherapy, relevant to the presence of cancer, relevant to the presence of infection (such as, for example HIV infection)) and metabolic syndrome. In some embodiments, the conditions associated with hypogonadism are those listed in the OMIM database and include, but are not limited to, gene mutations or conditions associated with: hypogonadism with mild intellectual disability and microcephaly (OMIM: 241000), cerebellar ataxia and hypergonadotropic hypogonadism (OMIM: 605672), congenital muscular dystrophy with incipient cataracts and hypogonadism (OMIM: 254000), hypogonadism-cataract syndrome (OMIM: 240950), with intellectual disability and Male hypogonadism with skeletal abnormalities (OMIM: 307500), Hypogonadism, including male hypogonadism and testicular atrophy (OMIM: 241100), Retinitis pigmentosa, deafness, intellectual disability, and hypogonadism (OMIM: 268020), Deafness-hypogonadism syndrome (OMIM: 304350), Polynuclear myopathy with intellectual disability, short stature, and hypogonadism (OMIM: 253320), Hypergonadotropic hypogonadism and focal alopecia (OMIM: 241090),Alopecia-mental retardation syndrome with convulsions and hypergonadotropic hypogonadism (OMIM: 601217), Johnson neuroectodermal syndrome (OMIM: 147770), Hypogonadotropic type 9 with or without anosmia; HH9 cytogenetic location: 9q34.3 (OMIM: 614838), Hypergonadotropic type 6 with or without anosmia; HH6 cytogenetic location: 10q24.32 (OMIM: 612702), including ichthyosis and male hypogonadism Rudd syndrome Syndrome (OMIM: 308200), hypogonadotropic type 5 with or without anosmia; HH5 cytogenetic location: 8q12.2 (OMIM: 612370), hypergonadotropic hypogonadism type 19 with or without anosmia; HH19 cytogenetic location: 12q21.33 (OMIM: 615269), hypergonadotropic hypogonadism type 17 with or without anosmia; HH17 cytogenetic location: 5q31.3 (OMIM: 615266), with or without Hypergonadotropic hypogonadism 20 with anosmia; HH20 cytogenetic location: 8p21.3 (OMIM: 615270), hypergonadotropic hypogonadism type 4 with or without anosmia; HH4 cytogenetic location: 3p13 (OMIM: 610628), hypergonadotropic hypogonadism type 21 with or without anosmia; HH21 cytogenetic location: 20p12.1 (OMIM: 615271), hypergonadotropic hypogonadism type 10 with or without anosmia; H H10 cytogenetic location: 12q13.3 (OMIM: 614839), dilated cardiomyopathy with hypergonadotropic hypogonadism (OMIM: 212112), hypergonadotropic hypogonadism type 11 with or without anosmia; HH11 cytogenetic location: 4q24 (OMIM: 614840), hypergonadotropic hypogonadism type 3 with or without anosmia; HH3 cytogenetic location: 20p12.3 (OMIM: 244200), carcinoid tumor syndrome (scholte syndrome); SHLTS (OMIM: 300977), hypergonadotropic hypogonadism type 18 with or without anosmia; HH18 cytogenetic location: 3p14.3 (OMIM: 615267), hypergonadotropic hypogonadism type 13 with or without anosmia; HH13 cytogenetic location: 1q32.1 (OMIM: 614842), hypergonadotropic hypogonadism type 7 with or without anosmia; HH7 cytogenetic location: 4q13.2 (OMIM: 146110), with short stature,Moyamoya disease type 4 with hypergonadotropic hypogonadism and facial dysmorphism; MYMY4 cytogenetic location: Xq28 (OMIM: 300845), hypomyelinating leukodystrophy type 7 with or without oligodontia and / or hypergonadotropic hypogonadism; HLD7 cytogenetic location: 10q22.3 (OMIM: 607694), Woodhouse-Sakati syndrome; cytogenetic location: 2q31.1 (OMIM: 241080), hypergonadotropic hypogonadism type 14 with or without anosmia; HH14 cytogenetic location: 10q26.12 (OMIM: 614858), Gordon Holmes syndrome; GDHS cytogenetic location: 7p22.1 (OMIM: 212840), hypomyelinating leukodystrophy type 8 with oligodontia and / or hypergonadotropic hypogonadism; HLD8 cytogenetic location: 12q23.3 (OMIM: 614381), hypergonadotropic hypogonadism type 12 with or without anosmia; HH12 cytogenetic location: 8p21.2 (OMIM: 614841), Martsolf Syndrome; Cytogenetic location: 1q41 (OMIM: 212720), Boucher-Neuhauser syndrome; BNHS Cytogenetic location: 19p13.2 (OMIM: 215470), Hypergonadotropic hypogonadism type 15 with or without anosmia; HH15 Cytogenetic location: 2q14.3 (OMIM: 614880), Hypergonadotropic hypogonadism type 22 with or without anosmia; HH22 Cytogenetic location: 2q14.3 (OMIM: 614880), Cytogenetic location: 7q31.32 (OMIM: 616030), hypergonadotropic hypogonadism type 1 with or without anosmia; HH1 cytogenetic location: Xp22.31 (OMIM: 308700), hypergonadotropic hypogonadism type 24 without anosmia; HH24 cytogenetic location: 11p14.1 (OMIM: 229070), Bosma anorhinal microphthalmia syndrome; BAMS cytogenetic location: 18p11.32 (OMIM: 229070), Bosma anorhinal microphthalmia syndrome OMIM: 603457), including Leydig cell dysgenesis type I and Leydig cell dysgenesis type II; cytogenetic location: 2p16.3 (OMIM: 238320), hypergonadotropic hypogonadism type 16 with or without anosmia; HH16 cytogenetic location: 7q21.11 (OMIM: 614897), mental retardation syndrome X-linked Cabezas type; MRXSC cytogenetic location: Xq24 (OMIM: 300354),Hypergonadotropic hypogonadism type 23 without anosmia; HH23 cytogenetic location: 19q13.33 (OMIM: 228300), Hypergonadotropic hypogonadism type 8 with or without anosmia; HH8 cytogenetic location: 19p13.3 (OMIM: 614837), Adrenal hypoplasia congenita; AHC cytogenetic location: Xp21.2 (OMIM: 300200), Combined pituitary hormone deficiency type 2; CPHD2 cytogenetic location: 5q35.3 (OMIM: 262600), Hypergonadotropic hypogonadism type 2 with or without anosmia; HH2 cytogenetic location: 8p11. 23 (OMIM: 147950), Na+ / K+ interacting transporting ATPase 2, including the NKAIN2TCBA1 / SUSP1 fusion gene; cytogenetic location: 6q22.31 (OMIM: 609758), MEHMO syndrome; MEHMO cytogenetic location: 1pter-p36.13, Xp22.11 (OMIM: 300148), histiocytosis-lymphadenopathy syndrome; cytogenetic location: 1pter-p36.13, 10q22.1 (OMIM: 602782), Waardenburg syndrome type 2E; WS2E cytogenetic location: 22q13.1 (OMIM: 611584 ), Brachydactyly with characteristic facies and Kallmann syndrome (OMIM: 113480), Testes, Rudimentary (OMIM: 273150), Mental retardation syndrome Belgian type (OMIM: 249599), Spastic paralysis and deafness (OMIM: 312910), Tachykinin receptor 3; TACR3 cytogenetic location: 4q24 (OMIM: 162332), NMDA receptor synaptic nuclear signaling and neuronal migration factor; NSMF cytogenetic location: 9q34.3 (OMIM: 608137), Tachykinin 3; TAC3 cytogenetic location: 12q13.3 ( OMIM: 162330), RNA polymerase III subunit B; POLR3B cytogenetic location: 12q23.3 (OMIM: 614366), KISS1 receptor; KISS1R cytogenetic location: 19p13.3 (OMIM: 604161), RNA polymerase III subunit A; POLR3A cytogenetic location: 10q22.3 (OMIM: 614258), SPROUTY, Drosophila homolog type 4; SPRY4 cytogenetic location: 5q31.3 (OMIM: 607984), WD repeat protein 11; WDR11 cytogenetic location: 10q26.12 (OMIM: 606417),Interleukin 17 receptor D; IL17RD cytogenetic location: 3p14.3 (OMIM: 606807), fibronectin-like domain-containing leucine-rich transmembrane protein 3; FLRT3 cytogenetic location: 20p12.1 (OMIM: 604808), gonadotropin-releasing hormone receptor; GNRHR cytogenetic location: 4q13.2 (OMIM: 138850), prokineticin type 2; PROK2 cytogenetic location: 3p13 (OMIM: 607002), dual-specificity phosphatase 6; DUSP6 cytogenetic location: 12q21.33 (OMIM: 602748), cone-rod dystrophy type 1; CO RD1 cytogenetic location: 18q21.1-q21.3 (OMIM: 600624), fibroblast growth factor 17; FGF17 cytogenetic location: 8p21.3 (OMIM: 603725), Fez family zinc finger protein 1; FEZF1 cytogenetic location: 7q31.32 (OMIM: 613301), heparan sulfate 6-O-sulfate triphosphatase 1; HS6ST1 cytogenetic location: 2q14.3 (OMIM: 604846), KAL1 gene; KAL1 cytogenetic location: Xp22.31 (OMIM: 300836), conductive hearing loss with external ear malformation (OMIM: 221300), follicle-stimulating hormone , beta polypeptide; FSHB cytogenetic location: 11p14.1 (OMIM: 136530), hemochromatosis type 2B; HFE2B cytogenetic location: 19q13.12 (OMIM: 613313), fibroblast growth factor receptor 1; including FGFR1FGFR1 / BCR fusion gene; cytogenetic location: 8p11.23 (OMIM: 136350), luteinizing hormone, beta polypeptide; LHB cytogenetic location: 19q13.33 (OMIM: 152780), prokineticin receptor 2; PROKR2 cytogenetic location: 20p12.3 (OMIM: 607123), KISS1 metastasis suppressor; KISS 1 Cytogenetic location: 1q32.1 (OMIM: 603286), gonadotropin-releasing hormone 1, GNRH1 prolactin release inhibitory factor, including PIF; Cytogenetic location: 8p21.2 (OMIM: 152760), obesity-hypogonadism-mental disability syndrome (OMIM: 601794), Bardet-Bieder syndrome type 19; BBS19 Cytogenetic location: 22q12.3 (OMIM: 615996), chromodomain helicase DNA binding protein 7; CHD7 Cytogenetic location: 8q12.2 (OMIM: 608892), Kallmann syndrome with spastic paralysis (OMIM: 308750),Luteinizing hormone / chorionic gonadotropin receptor; LHCGR cytogenetic location: 2p16.3 (OMIM: 152790), semaphorin 3A; SEMA3A cytogenetic location: 7q21.11 (OMIM: 603961), Wilson-Turner X-linked mental retardation syndrome; WTS cytogenetic location: Xq12 (OMIM: 309585), fibroblast growth factor 8; FGF8 cytogenetic location: 10q24.32 (OMIM: 600483), Bardet-Biedl syndrome type 11; BBS11 cytogenetic location: 9q33.1 (OMIM: 615988), BARDE T-BIEDL syndrome type 20; BBS20 Cytogenetic location: 9p21.2 (OMIM: 617119), Microcephaly with chemotaxis defect and transient hypogammaglobulinemia (OMIM: 251240), Richards-Rundle syndrome; RRNS (OMIM: 245100), Limb defects, ectodermal dysplasia, ear malformations, and other anomalies (OMIM: 273400), Hemophilia type 2A; includes HFE2A Hemochromatosis type 2; includes HFE2; Cytogenetic location: 1q21.1 (OMIM: 602390), Mandibular dysplasia, deafness, progeria, and lipodystrophy syndrome; M DPL cytogenetic location: 19q13.33 (OMIM: 615381), spinocerebellar ataxia, autosomal recessive 16; SCAR16 cytogenetic location: 16p13.3 (OMIM: 615768), progressive external ophthalmoplegia with mitochondrial DNA deletion, autosomal dominant 1; PEOA1 cytogenetic location: 15q26.1 (OMIM: 157640), leukoencephalopathy with dystonia and motor neuropathy; LKDMN cytogenetic location: 1p32.3 (OMIM: 613724), Bardet-Biedl syndrome type 8; BBS8 cytogenetic location: 14q31.3 (OMIM: 615768), progressive external ophthalmoplegia with mitochondrial DNA deletion, autosomal dominant 1 IM: 615985), Bjornstad syndrome; BJS cytogenetic location: 2q35 (OMIM: 262000), Marinesco-Sjogren syndrome; MSS cytogenetic location: 5q31.2 (OMIM: 248800), partial androgen insensitivity; PAIS cytogenetic location: Xq12 (OMIM: 312300), ichthyosis X-linked, including complex XLI ichthyosis X-linked; cytogenetic location: xp22.31 (OMIM: 308100), Bardet-Biedl syndrome type 5; BBS5 cytogenetic location: 2q31.1 (OMIM: 615983),Acrometropia Demirhan type; AMDD cytogenetic location: 4q22.3 (OMIM: 609441), Nephrotic syndrome 14; NPHS14 cytogenetic location: 10q22.1 (OMIM: 617575), Intellectual disability-facial hypotonia syndrome X-linked type 1; MRXHF1 cytogenetic location: Xq21.1 (OMIM: 309580), Dilanger syndrome type 5; CDLS5 cytogenetic location: Xq13.1 (OMIM: 300882), 114.#612079 - Alopecia, neurologic deficits, and endocrinopathy syndrome; ANES cytogenetic location: 7q32.1 (OMIM: 617575). IM: 612079), RING finger protein 216; RNF216 cytogenetic location: 7p22.1 (OMIM: 609948), leptin deficiency or dysfunction; LEPD cytogenetic location: 7q32.1 (OMIM: 614962), Bardet-Biedl syndrome type 17; BBS17 cytogenetic location: 3p21.31 (OMIM: 615994), OLIVER-MCFARLANE syndrome; OMCS cytogenetic location: 19p13.2 (OMIM: 275400), Moebius syndrome; MBS cytogenetic location: 13q12.2-q13 (OMIM: 157900), Bardet-Biedl syndrome type 12; BBS12 cytogenetic location: 4q27 (OMIM: 615989), Microphthalmia syndrome type 3, including MCOPS3 optic nerve hypoplasia and central nervous system abnormalities; cytogenetic location: 3q26.33 (OMIM: 206900), Chromosome Xq27.3-q28 duplication syndrome; cytogenetic location: Xq27.3-q28 (OMIM: 300869), Alström syndrome; ALMS cytogenetic location: 2p13.1 (OMIM: 203800), Galactosemia, including Duarte variant; cytogenetic location: 9p13.3 (OMIM: 23 0400), combined oxidative phosphorylation deficiency type 34; COXPD34 cytogenetic location: 17q25.1 (OMIM: 617872), Bardet-Biedl syndrome type 1; BBS1 cytogenetic location: 1p35.2, 1pter-p36.13, 3q11.2 (OMIM: 209900), congenital cataract, facial dysmorphism, and neuropathy; CCFDN cytogenetic location: 18q23 (OMIM: 604168), microcytic hypochromic anemia with iron excess type 2; AHMIO2 cytogenetic location: 2q14.2 (OMIM: 615234), Prader-Willi syndrome,The PWS Prader-Willi syndrome chromosomal regions included are: PWCR cytogenetic location: 1pter-p36.13, 15q11.2 (OMIM: 176270), hemochromatosis type 1; HFE1 cytogenetic location: 1pter-p36.13, 20p12.3 (OMIM: 235200), Senior-Loken syndrome type 9; SLSN9 cytogenetic location: 2q37.3 (OMIM: 616629), mitochondrial DNA depletion syndrome type 11; MTDPS11 cytogenetic location: 20p11.23 (OMIM: 615084), α-methylacyl-CoA depletion syndrome type 11. Gyrase deficiency; AMACRD cytogenetic location: 5p13.2 (OMIM: 614307), Sifrim-Hitz-Weiss syndrome; SIHIWES cytogenetic location: 12p13.31 (OMIM: 617159), congenital disorder of glycosylation type Ik; CDG1K cytogenetic location: 16p13.3 (OMIM: 608540), congenital disorder of glycosylation type It; CDG1T cytogenetic location: 1p31.3 (OMIM: 614921), multiple endocrine neoplasia-polyneuropathy syndrome; PEPNS cytogenetic location: 15q21.2 (OMIM: 616113 ), proprotein convertase 1 / 3 deficiency; cytogenetic location: 5q15 (OMIM: 600955), Bardet-Biedl syndrome type 2; BBS2 cytogenetic location: 16q13 (OMIM: 615981), MACS syndrome; genetic location: 20p11.23 (OMIM: 613075), mitochondrial DNA depletion syndrome type 7 (hepatocerebral type); MTDPS7 cytogenetic location: 10q24.31 (OMIM: 271245), microcephaly, epilepsy, and diabetes syndrome; MEDS cytogenetic location: 18q21.1 (OMIM: 614231), Culler-J ones syndrome; CJS cytogenetic location: 2q14.2 (OMIM: 615849), Schaaf-Yang syndrome; SHFYNG cytogenetic location: 15q11.2 (OMIM: 615547), hypotonia-cystinuria syndrome, including homozygous 2p21 deletion syndrome; cytogenetic location: 2p21 (OMIM: 606407), autoimmune polyendocrine syndrome type I with or without reversible metaphyseal dysgenesis; including APS1 autoimmune polyendocrine syndrome type I, autosomal dominant inheritance, cytogenetic location: 21q22.3 (OMIM: 240300),Rothmund-Thomson syndrome; RTS cytogenetic location: 8q24.3 (OMIM: 268400), Primrose syndrome; PRIMS cytogenetic location: 3q13.31 (OMIM: 259050), Chromosome 2p16.1-p15 deletion syndrome; cytogenetic location: 2p16.1-p15 (OMIM: 612513), Witteveen-Kolk syndrome; including WITKOS chromosome 15q24 deletion syndrome, including cytogenetic location: 15q24.2 (OMIM: 613406), Myotonic dystrophy type 2; DM2 cytogenetic location: 3q21.3 ... M: 602668), Fanconi anemia, complementation group A; FANCA Fanconi anemia, including the Estren-Dameshek variant, cytogenetic location: 16q24.3 (OMIM: 227650), Congenital disorder of glycosylation type Ia; CDG1A cytogenetic location: 16p13.2 (OMIM: 212065), Myotonic dystrophy type 1; DM1 cytogenetic location: 19q13.32 (OMIM: 160900), Congenital deafness with systemic albinism (OMIM: 220900), Hemochromatosis type 3; HFE3 cytogenetic location: 7q22.1 (OMIM: 604250), Camu rati-Engelmann disease type 2 (OMIM: 606631), Bardet-Biedl syndrome type 10; BBS10 cytogenetic location: 12q21.2 (OMIM: 615987), Bardet-Biedl syndrome type 16; BBS16 cytogenetic location: 1q43-q44 (OMIM: 615993), Perrault syndrome type 5; PRLTS5 cytogenetic location: 10q24.31 (OMIM: 616138), Fanconi anemia, complementation group E; FANCE cytogenetic location: 6p21.31 (OMIM: 600901), Bardet-Bi edl syndrome type 4; BBS4 cytogenetic location: 15q24.1 (OMIM: 615982), Waardenburg syndrome type 4C; WS4C cytogenetic location: 22q13.1 (OMIM: 613266), pseudohypoparathyroidism type IC; PHP1C cytogenetic location: 20q13.32 (OMIM: 612462) xeroderma pigmentosum, complementation group B; including XPB xeroderma pigmentosum B / Cockayne syndrome, cytogenetic location: 2q14.3 (OMIM: 610651), Fanconi anemia, complementation group D2; FANCD2 cytogenetic location: 3p25.3 (OMIM: 227646),Mirage syndrome; MIRAGE cytogenetic location: 7q21.2 (OMIM: 617053), Fanconi anemia, complementation group C; FANCC cytogenetic location: 9q22.32 (OMIM: 227645), acrodysplasia with or without steroid resistance type 1 ACRDYS1 cytogenetic location: 17q24.2 (OMIM: 101800), ectodermal dysplasia and cleft lip and palate syndrome type 1; EEC1 cytogenetic location: 7q11.2-q21.3 (OMIM: 227645). MIM: 129900), ectodermal dysplasia and cleft lip and palate syndrome type 3; EEC3 cytogenetic location: 3q28 (OMIM: 604292), chondrodysplasia punctata type 1, X recessive linked; CDPX1 cytogenetic location: Xp22.33 (OMIM: 302950), progressive external ophthalmoplegia with mitochondrial DNA deletion, autosomal dominant 3; PEOA3 cytogenetic location: 10q24.31 (OMIM: 609286), Cockayne syndrome type A; CSA cytogenetic location: 5q12.1 (OMIM: 216400), Werner syndrome; WRN cytogenetic location: 8p12 (OMIM: 277700), pseudohypoparathyroidism type IA; PHP1A cytogenetic location: 20q13.32 (OMIM: 103580), ataxia telangiectasia; included in complementation group A; includes ATA, cytogenetic location: 11q22.3 (OMIM: 208900), Noonan syndrome type 1; includes NS1 Pterygium syndrome, cytogenetic location: 12q24.13 (OMIM: 163950), OTU domain-containing protein 4; OTUD4 cytogenetic location: 4q31.21 (OMIM: 611744), DDB1 and CUL4-related factor 17; DCAF17 cytogenetic location: 2q31.1 (OMIM: 612515), nuclear receptor subfamily type 0, group B, number 1; NR0B1 cytogenetic location: Xp21.2 (OMIM: 300473), Cullin 4B; CUL4B cytogenetic location: Xq24 (OMIM: 300304), patatin-like phospholipase domain-containing protein 6; PNPLA6 cytogenetic location: 19p13.2 (OMIM: 603197), SRY box 2; SOX2 cytogenetic location: 3q26.33 (OMIM: 184429), PROP paired homeobox 1; PROP1 cytogenetic location: 5q35.3 (OMIM: 601538), semaphorin 3E; SEMA3E cytogenetic location: 7q21.11 (OMIM: 608166), congenital ovarian dysgenesis with short stature and recurrent metabolic acidosis,Hypergonadotropin (OMIM:605756), DMX class type 2; DMXL2 cytogenetic location: 15q21.2 (OMIM:612186), Chinese hamster X-ray repair, complement deficiency type 4; XRCC4 cytogenetic location: 5q14.2 (OMIM:194363), solute carrier family 29 (nucleoside transporter), number 3; SLC29A3 cytogenetic location: 10q22.1 (OMIM:612373), lamin A / C; LMNA including lamin A, cytogenetic location: 1q22 (OMIM:150330), Lin and Getting's craniosynostosis-mental disability syndrome (OMIM:2186 49), isolated congenital anosmia; ANIC cytogenetic location: 18p11.23-q12.2 (OMIM: 107200), Beamon syndrome type II (OMIM: 210350), zincemia with functional zinc depletion (OMIM: 601979), chromosome Xp21 deletion syndrome, cellular location: Xp21 (OMIM: 300679), helical domain protein 141; CCDC141 cytogenetic location: 2q31.2 (OMIM: 616031), Laurence-Moon syndrome; LNMS cytogenetic location: 19p13.2 (OMIM: 245800), congenital ovarian dysgenesis type 5; OD G5 cytogenetic location: 9q34.3 (OMIM: 617690), hypomyelinating leukodystrophy type 11; HLD11 cytogenetic location: 6p21.1 (OMIM: 616494), small nucleolar RNA, C / D box 116-1; SNORD116-1 cytogenetic location: 15q11.2 (OMIM: 605436), X-linked mental retardation, X-linked syndrome type 7; MRXS7 cytogenetic location: Xp11.3-q22 (OMIM: 300218), pineal hyperplasia, insulin-resistant diabetes mellitus, and physical abnormalities; cytogenetic location: 19p13.2 (OMIM: 262190), charge syndrome; Cytogenetic location: 1pter-p36.13, 7q21.11 (OMIM: 214800), Roifman syndrome; RFMN cytogenetic location: 2q14.2 (OMIM: 616651POLO-like kinase 4); PLK4 cytogenetic location: 4q28.1 (OMIM: 605031), aromatase excess syndrome; including familial AEXS gynecomastia due to increased aromatase activity; cytogenetic location: 15q21.2 (OMIM: 139300), Borjeson-Forssman-Lehmann syndrome; BFLS cytogenetic location: Xq26.2 (OMIM: 301900),Histone deacetylase 8; HDAC8 cytogenetic location: Xq13.1 (OMIM: 300269), Robinow syndrome, autosomal dominant 1; DRS1 cytogenetic location: 3p14.3 (OMIM: 180700), STIP1 homologous and U-box containing protein 1; STUB1 cytogenetic location: 16p13.3 (OMIM: 607207), microcephaly type 1, autosomal recessive; MCPH1 cytogenetic location: 8p23.1 (OMIM: 251200), aromatase deficiency cytogenetic location: 15q21.2 (OMIM: IM: 613546), Metaphyseal dysplasia, spondylosis, and hyperplasia (OMIM: 608811), Coenzyme Q10 deficiency type 1; COQ10D1 cytogenetic location: 4q21.22-q21.23 (OMIM: 607426), Retinopathy, pigmentation, and intellectual disability (OMIM: 268050), Chromosome structural maintenance protein containing flexible hinge domain 1; SMRHD1 cytogenetic location: 18p11.32 (OMIM: 614982), Cytochrome P450, family 19, subfamily A, polypeptide 1; includes CYP19A1 CYP19A1 / CGNL1 fusion gene, cytogenetic location: 15q21.2 (OMIM: 107910), N-myristoyltransferase 2; NMT2 cytogenetic location: 10p13 (OMIM: 603801), DNA polymerase gamma; POLG cytogenetic location: 15q26.1 (OMIM: 174763), leptin; LEP cytogenetic location: 7q32.1 (OMIM: 164160), cingulin-like 1; including CGNL1 CGNL1 / CYP19A1 fusion gene, cytogenetic location: 15q21.3 (OMIM: 607856), including BKMDNABKMA2, genetic location: 6q21 (OMIM: 109780), leptin receptor deficiency, genetic location: 1p31.3 (OMIM: 614963), Bardet-Biedl syndrome type 7; BBS7 cytogenetic location: 4q27 (OMIM: 615984), torticollis, keloids, cryptorchidism, and renal Dysgenesis; TKCR cytogenetic location: Xq28 (OMIM: 314300), TAX1-binding protein 3; TAX1BP3 cytogenetic location: 17p13.2 (OMIM: 616484), immunoglobulin superfamily, number 10; IGSF10 cytogenetic location: 3q25.1 (OMIM: 617351), short stature, microcephaly and endocrine dysfunction; SSMED cytogenetic location: 5q14.2 (OMIM: 616541),Perrault syndrome type 3; PRLTS3 cytogenetic location: 19p13.3 (OMIM: 614129), Hyperprolactinemia; HPRL cytogenetic location: 5p13.2 (OMIM: 615555), Chromosome 17q21.31 duplication syndrome; Cytogenetic location: 17q21.31 (OMIM: 613533), Premature ovarian failure 10; Quantitative trait locus for natural age-related menopause 3 including POF10; including MENOQ3, cytogenetic location: 20p12.3 (OMIM: 612885), BRCA1 / BRCA2 complex, subunit 3; BRCC3 cytogenetic location: Xq 28 (OMIM: 300617), large tumor suppressor, Drosophila homolog type 1; LATS1 cytogenetic location: 6q25.1 (OMIM: 603473), Warburg micro syndrome type 4; WARBM4 cytogenetic location: 20p13 (OMIM: 615663), mature T-cell proliferation 1; MTCP1 cytogenetic location: Xq28 (OMIM: 300116), 46, XX sex reversal 1; SRXX146, XX true hermaphroditism containing SRY-positive, cytogenetic location: Yp11.2 (OMIM: 400045), unilateral or bilateral cryptorchidism, cytogenetic location: 19p13.1 1 (OMIM: 219050), double sex and MAB3-related transcription factor 2; DMRT2 cytogenetic location: 9p24.3 (OMIM: 604935), zinc finger protein, X-linked; ZFX cytogenetic location: Xp22.11 (OMIM: 314980), non-photosensitive trichothiodystrophy type 4; TTD4 cytogenetic location: 7p14.1 (OMIM: 234050), Teashirt zinc finger homeobox type 1; TSHZ1 cytogenetic location: 18q22.3 (OMIM: 614427), limb-mammary syndrome; LMS cytogenetic location: 3q28 (OMIM: 603543), Ovarian dysplasia type 4; ODG4 cytogenetic location: 6q22.31 (OMIM: 616185), transporter receptor 2; TFR2 cytogenetic location: 7q22.1 (OMIM: 604720), 247. Early infantile epileptic encephalopathy type 28; EIEE28 cytogenetic location: 16q23.1-q23.2 (OMIM: 616211), Warburg micro syndrome type 3; WARBM3 cytogenetic location: 10p12.1 (OMIM: 614222), Hartsfield syndrome; HRTFDS cytogenetic location: 8p11.23 (OMIM: 615465),250.% 252350-Moyamoya disease type 1; MYMY1 cytogenetic location: 3p26-p24.2 (OMIM: 252350), hepcidin regulatory protein; HJV cytogenetic location: 1q21.1 (OMIM: 608374), Fanconi anemia, complementation group B; FANCB cytogenetic location: Xp22.2 (OMIM: 300514), intrauterine growth retardation, metaphyseal dysplasia and genital malformations; cytogenetic location: 11p15.4 (OMIM: 614732), RAB3 GTPase activating protein, nongenetic subunit; RAB3GAP2 cytogenetic location: 1q41 (OMIM: 609275), Carpenter syndrome type 1; CRPT1 cytogenetic location: 6p12.1-p11.2 (OMIM: 201000), phosphoglycerate dehydrogenase deficiency; PHGDHD cytogenetic location: 1p12 (OMIM: 601815), caseinolytic mitochondrial matrix peptidase proteolytic subunit; CLPP cytogenetic location: 19p13.3 (OMIM: 6011 19), 46, XY sex reversal 4; SRXY4 cytogenetic location: 9p24.3 (OMIM: 154230), leucine zipper transcription factor-like 1; LZTFL1 cytogenetic location: 3p21.31 (OMIM: 606568), pontocerebellar dysgenesis type 7; PCH7 cytogenetic location: 1p34.1 (OMIM: 614969), spermatogenesis impaired type 1; SPGF1 (OMIM: 258150), SIL1, S. cerevisiae homolog; SIL1 cytogenetic location: 5q31.2 (O MIM: 608005), 3MC syndrome type 1; 3MC1 cytogenetic location: 3q27.3 (OMIM: 257920), lipedema (OMIM: 614103), inhibitor, βB; including INHBB activator β-B, cytogenetic location: 2q14.2 (OMIM: 147390), DNA methyltransferase 3-like protein; DNMT3L cytogenetic location: 21q22.3 (OMIM: 606588), Necdin; NDN cytogenetic location: 15q11.2 (OMIM :602117), Hypoparathyroidism-retardation-malformation syndrome; HRDS cytogenetic location: 1q42.3 (OMIM: 241410), Optic atrophy with or without deafness, ophthalmoplegia, myopathy, ataxia, and neuropathy; cytogenetic location: 3q29 (OMIM: 125250), 46,XX sex reversal type 4; SRXX4 cytogenetic location: 9q33.3 (OMIM: 617480), Cerebral-ocular dwarfism, cytogenetic location: 17q22 (OMIM: 253250),Microphthalmia syndrome type 6; MCOPS6 cytogenetic location: 14q22.2 (OMIM: 607932), PHD finger protein 6; PHF6 cytogenetic location: xq26.2 (OMIM: 300414), proprotein convertase, subtilisin / kexin type 1; PCSK1 cytogenetic location: 5q15 (OMIM: 162150), spherocytosis type 1; SPH1 cytogenetic location: 8p11.21 (OMIM: 182900), GAPO syndrome, cytogenetic location: 2p13.3 (OMIM: 230740), inhibin, beta A; including INHBA activating protein A, cytogenetic location: 7p14.1 (OMIM: 182900), IM: 147290), SRY-box 10; SOX10 cytogenetic location: 22q13.1 (OMIM: 602229), Progressive external ophthalmoplegia with mitochondrial DNA deletion, autosomal recessive 1; PEOB1 cytogenetic location: 15q26.1 (OMIM: 258450), Hormone-sensitive lipase; Adipogenetic cytogenetic location: 19q13.2 (OMIM: 151750), Minichromosome maintenance (MCM) complex type 8; MCM8 cytogenetic location: 20p12.3 (OMIM: 608187), FANCM gene; FANCM cytogenetic location: 14q21.2 (OMIM: 609644), Multiple Pterygium syndrome, Escobar variant; EVMPS cytogenetic location: 2q37.1 (OMIM: 265000), Warburg micro syndrome type 1; WARBM1 cytogenetic location: 2q21.3 (OMIM: 600118), trichophalangeal syndrome type I; TRPS1 cytogenetic location: 8q23.3 (OMIM: 190350), microcephalic dysostotic primordial dwarfism type I; MOPD1 cytogenetic location: 2q14.2 (OMIM: 210710), follicle-stimulating hormone receptor; FSHR cytogenetic location: 2p16.3 (OMIM: 136435), Mage-like syndrome type 2; MAGEL2 Cytogenetic location: 15q11.2 (OMIM: 605283), Norrie disease; ND cytogenetic location: Xp11.3 (OMIM: 310600), ATP-dependent DNA ligase; LIG4 cytogenetic location: 13q33.3 (OMIM: 601837), superoxide dismutase 2; SOD2 cytogenetic location: 6q25.3 (OMIM: 147460), Hutchinson-Gilford progeria syndrome; including HGPS progeria syndrome childhood onset, (OMIM: 176670), combined pituitary hormone deficiency type 1; CPHD1 cytogenetic location: 3p11.2 (OMIM: 613038),ALMS1 gene; ALMS1 cytogenetic location: 2p13.1 (OMIM: 606844), photosensitive trichothiodystrophy type 1; TTD1 cytogenetic location: 19q13.32 (OMIM: 601675), including obesity and thinness, cytogenetic location: 5q13.2, 1pter-p36.13, 4q31.1, 1pter-p36.13, 3p25.2, 1pter-p36.13, 3p25.3, 1pter-p36.13, 1p36.11, 1pter-p36.13, 2p23.3, 1pter-p36.13, 18q21.32, 1pter-p36 .13, 16q22.1 (OMIM: 601665), linear cutis defect type 1 with multiple congenital malformations; LSDMCA1, cytogenetic location: Xp22.2 (OMIM: 309801), galanin; GAL, cytogenetic location: 11q13.2 (OMIM: 137035), Jacobsen syndrome; JBS, cytogenetic location: 11q23 (OMIM: 147791), WW domain-containing oxidoreductase; including WWOX fragile site 16q23.2; including FRA16D, cytogenetic location: 16q23.1-q23.2 (OMIM: 605131), FANCG gene; FA NCG cytogenetic location: 9p13.3 (OMIM: 602956), steroid sulfatase; includes STS steroid sulfatase isozyme S, cytogenetic location: Xp22.31 (OMIM: 300747), hepcidin antimicrobial peptide; HAMP cytogenetic location: 19q13.12 (OMIM: 606464), leptin receptor; LEPR cytogenetic location: 1p31.3 (OMIM: 601007), Kearns-Sayre syndrome; KSS (OMIM: 530000), cystinosis, nephrotic; CTNS cystinosis, infantile nephrotic, includes cytogenetic location: 17p13.2 (OMIM: 530000), cystinosis, nephrotic IM: 219800), Wolfram syndrome 1; WFS1 cytogenetic location: 4p16.1 (OMIM: 222300), Chinese hamster excision repair, complement deficiency type 5; ERCC5 cytogenetic location: 13q33.1 (OMIM: 133530), Hirschsprung's disease type 1 susceptible; including HSCR1 preventive Hirschsprung's disease, cytogenetic location: 10q11.21 (OMIM: 142623), androgen receptor; AR cytogenetic location: Xq12 (OMIM: 313700), Spinocerebellar ataxia type 6; SCA6 cytogenetic location: 19p13.13 (OMIM: 183086),312.#106210 - Aniridia type 1; includes AN1 congenital cataract with delayed corneal dystrophy, cytogenetic location: 11p13 (OMIM: 106210), FANCA gene; FANCA cytogenetic location: 16q24.3 (OMIM: 607139), nuclear receptor subfamily 5, group A, number 1; NR5A1 cytogenetic location: 9q33.3 (OMIM: 184757), melanocortin receptor 4; MC4R cytogenetic location: 18q21.32 (OMIM: 155541), tumor protein p63; TP63 cytogenetic location: 3q28 (OMIM: 6032 73), adrenoleukodystrophy; including ALD adrenomyeloneuropathy; AMN, including cytogenetic location: xq28 (OMIM: 300100), cerebellar ataxia and ectodermal dysplasia (OMIM: 212835), nescient helix-loop-helix 2; NHLH2 cytogenetic location: 1p13.1 (OMIM: 162361), spermatogenesis- and oogenesis-specific basic helix-loop-helix protein 1; SOHLH1 cytogenetic location: 9q34.3 (OMIM: 610224), pseudovagina-perineum-hypospadias; including PPSH short penis, cytogenetic location: 2p23 .1 (OMIM: 264600); Adrenal hyperplasia due to isolated 17α-hydroxylase deficiency / 17,20-lyase deficiency including cytogenetic location: 10q24.32 (OMIM: 202110), Eukaryotic translation initiation factor 2, subunit 3; EIF2S3 cytogenetic location: Xp22.11 (OMIM: 300161), Sphingosine-1-phosphate lyase 1; SGPL1 cytogenetic location: 10q22.1 (OMIM: 603729), 17β-hydroxysteroid dehydrogenase type III deficiency Polycystic ovary disease due to 17-ketosteroid reductase deficiency including cytogenetic location: 9q2 2.32 (OMIM: 264300), premature ovarian failure type 1; POF1 cytogenetic location: Xq27.3 (OMIM: 311360), 46, XY sex reversal 1; includes SRXY146, SRY-related XY true hermaphroditism, cytogenetic location: yp11.2 (OMIM: 400044), McCune-Albright syndrome; includes multiple osteofibrous dysplasia; includes PFD; includes POFD, cytogenetic location: 20q13.32 (OMIM: 174800) and / or ATR-X gene; ATRX cytogenetic location: Xq21.1 (OMIM: 300032). ,

[0124] In the therapeutic applications disclosed herein, administration may be in vitro (by contacting a peptide compound or a combination of peptide compounds with cells in culture) or in vivo (by administering a peptide compound or a combination of peptide compounds to a subject comprising cells). The subject may be a mammal, and in a further embodiment, may be a male. In one embodiment, the male is at least 30 years old, or in a further embodiment, at least 50 years old. After the age of 30, male testosterone production declines, with total testosterone levels declining by 1-2% per year. Therefore, testosterone replacement therapy (in this case, inducing endogenous T production) can be applied at any time when testosterone begins to decline and / or symptoms associated with testosterone decline (low libido and erection, low lean body mass, reduced energy, central obesity, lack of stressor response, etc. indicate testosterone decline). These symptoms are more significant with aging and are more common in men over 50, with cardiovascular disease, metabolic syndrome, and depression being added to the phenotypic list associated with testosterone decline. In addition, even at an age of less than 30 years old, the use of therapeutic agents described herein can be helpful in the case of male infertility due to hypogonadism.

[0125] In the therapeutic applications described herein, the treated cells may be from or located in the testis, and in further embodiments, the treated cells may be Leydig cells. In another embodiment, the cells may be from or located in the ovary, adrenal gland, and / or brain.

[0126] Therapeutic applications described herein can be used to prevent, treat and / or alleviate the symptoms of a disease associated with a decline in steroid levels. An exemplary disease associated with a decline in steroid levels is hypogonadism. Such diseases include, but are not limited to, infertility, aging, decreased libido, sexual dysfunction, mood changes, fatigue, decreased lean body mass, decreased bone mineral density, increased visceral fat, emaciation or metabolic syndrome.

[0127] Alternatively or in combination, the therapeutic applications described herein can be used to prevent, treat and / or alleviate symptoms associated with disorders associated with decreased steroid levels (e.g., decreased neurosteroid levels). Such disorders include, but are not limited to, anxiety disorders and depression, such as, for example, post-traumatic stress disorder.

[0128] Alternatively or in combination, the therapeutic applications described herein can be used to prevent, treat and / or alleviate symptoms associated with disorders associated with a decrease in steroid levels. Such disorders include, but are not limited to, depression, organ failure, myocardial stiffness, low energy, abnormal hematocrit, sepsis, and coping with stressors.

[0129] In the context of the present disclosure, a peptide compound or combination is provided to a subject (such as a mammal, including a human) in a pharmaceutically or therapeutically effective amount. The expressions "pharmaceutically effective amount" and "therapeutically effective amount" collectively refer to an amount (dose) effective in mediating a therapeutic benefit (e.g., preventing, treating and / or alleviating the symptoms of hypogonadism) to a subject. It should also be understood herein that a "pharmaceutically effective amount" can be interpreted as an amount that gives the desired therapeutic effect, taken in one dose or in any dose or route, alone or in combination with other therapeutic agents.

[0130] The therapeutically effective amount or dosage of the peptide compounds of the present disclosure may be in the range of about 0.001 to 1000 μg / kg body weight, other ranges of the invention include about 0.001 to 900 μg / kg body weight, about 0.001 to 800 μg / kg body weight, about 0.001 to 700 μg / kg body weight, about 0.001 to 600 μg / kg body weight, about 0.001 to 500 μg / kg body weight, about 0.001 to 450 μg / kg body weight, about 0.001 to 400 μg / kg body weight, about 0.001 to 350 μg / kg body weight, about 0.001 to 300 μg / kg body weight, about 0.001 to 250 μg / kg body weight, about 0.001 to 200 μg / kg body weight. , about 0.001 to 150 μg / kg body weight, about 0.001 to 100 μg / kg body weight, about 0.001 to 90 μg / kg body weight, about 0.001 to 80 μg / kg body weight, about 0.001 to 70 μg / kg body weight, about 0.001 to 60 μg / kg body weight, about 0.001 to 50 μg / kg body weight, about 0.001 to 40 μg / kg body weight, about 0.001 to 30 μg / kg body weight, about 0.001 to 20 μg / kg body weight, about 0.001 to 10 μg / kg body weight, about 0.01 to 900 μg / kg body weight, about 0.01 to 800 μg / kg body weight, about 0.01 to 700 μg / kg body weight, about 0.01 to 600 μg / kg body weight, about 0.01 to 500 μg / kg body weight, about 0.01 to 450 μg / kg body weight, about 0.01 to 400 μg / kg body weight, about 0.01 to 350 μg / kg body weight, about 0.01 to 300 μg / kg body weight, about 0.01 to 250 μg / kg body weight, about 0.01 to 200 μg / kg body weight, about 0.01 to 150 μg / kg body weight, about 0.01 to 100 μg / kg body weight, about 0.01 to 90 μg / kg body weight, about 0.01 to 80 μg / kg body weight, about 0.01 to 70 μg / kg body weight, about 0.01 to 60 μg / kg body weight, about 0.01 to 50 μg / kg body weight, about 0.01 to 40 μg / kg body weight. g / kg body weight, about 0.01 to 30 μg / kg body weight, about 0.01 to 20 μg / kg body weight, about 0.01 to 10 μg / kg body weight, about 0.1 to 900 μg / kg body weight, about 0.1 to 800 μg / kg body weight, about 0.1 to 700 μg / kg body weight, about 0.1 to 600 μg / kg body weight, about 0.1 to 500 μg / kg body weight, about 0.1 to 450 μg / kg body weight, about 0.1 to 400 μg / kg body weight, about 0.1 to 350 μg / kg body weight, about 0.1 to 300 μg / kg body weight, about 0.1 to 250 μg / kg body weight, about 0.1 to 200 μg / kg body weight, about 0.1 to 150 μg / kg body weight, about 0.1 to 100 μg / kg body weight, about 0.1 to 90 μg / kg body weight, about 0.1 to 80 μg / kg body weight, about 0.1 to 70 μg / kg body weight, about 0.1 to 60 μg / kg body weight, about 0.1 to 50 μg / kg body weight, about 0.1 to 40 μg / kg body weight, about 0.1 to 30 μg / kg body weight, about 0.1 to 20 μg / kg body weight, about 0.1 to 10 μg / kg body weight, about 1 to 900 μg / kg body weight, about 1 to 800 μg / kg body weight, about 1 to 700 μg / kg body weight, about 1 to 600 μg / kg body weight, about 1 to 500 μg / kg body weight, about 1 to 450 μg / kg body weight, about 1 to 400 μg / kg body weight, about 1 to 350 μg / kg body weight, about 1 to 300 μg / kg body weight, about 1 to 250 μg / kg body weight, about 1 to 200 μg / kg body weight, about 1 to 150 μg / kg body weight, about 1 to 100 μg / kg body weight, about 1 to 90 μg / kg body weight, about 1 to 80 μg / kg body weight, about 1 to 70 μg / kg body weight, about 1 to 60 μg / kg body weight, about 1 to 50 μg / kg body weight, about 1 to 40 μg / kg body weight, about 1 to 30 μg / kg body weight, about 1 to 20 μg / kg body weight, or about 1 to 10 μg / kg body weight.

[0131] When more than one peptide compound is used, the therapeutically effective amount of the total weight of the peptide compounds may be in the range of about 0.001 to 1000 μg / kg body weight, other ranges of the invention include about 0.001 to 900 μg / kg body weight, about 0.001 to 800 μg / kg body weight, about 0.001 to 700 μg / kg body weight, about 0.001 to 600 μg / kg body weight, about 0.001 to 500 μg / kg body weight, about 0.001 to 450 μg / kg body weight, about 0.001 to 400 μg / kg body weight, about 0.001 to 350 μg / kg body weight, about 0.001 to 300 μg / kg body weight, about 0.001 to 250 μg / kg body weight, about 0.001 to 300 μg / kg body weight, about 0.001 to 450 μg / kg body weight, about 0.001 to 400 μg / kg body weight, about 0.001 to 500 μg / kg body weight, about 0.001 to 600 μg / kg body weight, about 0.001 to 700 μg / kg body weight, about 0.001 to 800 μg / kg body weight, about 0.001 to 900 μg / kg body weight, about 0.001 to 1000 μg / kg body weight, about 0.001 to 1100 μg / kg body weight, about 0.001 to 1200 μg / kg body weight, about 0.001 to 1300 μg / kg body weight, about 0.001 to 1400 μg / kg body weight, about 0.001 to 1500 μ 1 to 200 μg / kg body weight, about 0.001 to 150 μg / kg body weight, about 0.001 to 100 μg / kg body weight, about 0.001 to 90 μg / kg body weight, about 0.001 to 80 μg / kg body weight, about 0.001 to 70 μg / kg body weight, about 0.001 to 60 μg / kg body weight, about 0.001 to 50 μg / kg body weight, about 0.001 to 40 μg / kg body weight, about 0.001 to 30 μg / kg body weight, about 0.001 to 20 μg / kg body weight, about 0.001 to 10 μg / kg body weight, about 0.01 to 900 μg / kg body weight, about 0.01 to 800 μg / kg body weight, about 0.01 to 700 μg / kg g body weight, about 0.01 to 600 μg / kg body weight, about 0.01 to 500 μg / kg body weight, about 0.01 to 450 μg / kg body weight, about 0.01 to 400 μg / kg body weight, about 0.01 to 350 μg / kg body weight, about 0.01 to 300 μg / kg body weight, about 0.01 to 250 μg / kg body weight, about 0.01 to 200 μg / kg body weight, about 0.01 to 150 μg / kg body weight, about 0.01 to 100 μg / kg body weight, about 0.01 to 90 μg / kg body weight, about 0.01 to 80 μg / kg body weight, about 0.01 to 70 μg / kg body weight, about 0.01 to 60 μg / kg body weight, about 0.01 to 50 μg / kg body weight, about 0.01 to 40 μg / kg body weight, about 0.01 to 30 μg / kg body weight, about 0.01 to 20 μg / kg body weight, about 0.01 to 10 μg / kg body weight, about 0.1 to 900 μg / kg body weight, about 0.1 to 800 μg / kg body weight, about 0.1 to 700 μg / kg body weight, about 0.1 to 600 μg / kg body weight, about 0.1 to 500 μg / kg body weight, about 0.1 to 450 μg / kg body weight, about 0.1 to 400 μg / kg body weight, about 0.1 to 350 μg / kg body weight, about 0.1 to 300 μg / kg body weight, about 0.1 to 250 μg / kg body weight, about 0.1 to 200 μg / kg body weight, about 0.1 to 150 μg / kg body weight, about 0.1 to 100 μg / kg body weight, about 0.1 to 90 μg / kg body weight, about 0.1 to 80 μg / kg body weight, about 0.1 to 70 μg / kg body weight, about 0.1 to 60 μg / kg body weight, about 0.1 to 50 μg / kg body weight, about 0.1 to 40 μg / kg body weight, about 0.1 to 30 μg / kg body weight, about 0.1 to 20 μg / kg body weight, about 0.1 to 10 μg / kg body weight, about 1 to 900 μg / kg body weight, about 1 to 800 μg / kg body weight, about 1 to 700 μg / kg body weight, about 1 to 600 μg / kg body weight, about 1 to 500 μg / kg body weight about 1 to 450 μg / kg body weight, about 1 to 400 μg / kg body weight, about 1 to 350 μg / kg body weight, about 1 to 300 μg / kg body weight, about 1 to 250 μg / kg body weight, about 1 to 200 μg / kg body weight, about 1 to 150 μg / kg body weight, about 1 to 100 μg / kg body weight, about 1 to 90 μg / kg body weight, about 1 to 80 μg / kg body weight, about 1 to 70 μg / kg body weight, about 1 to 60 μg / kg body weight, about 1 to 50 μg / kg body weight, about 1 to 40 μg / kg body weight, about 1 to 30 μg / kg body weight, about 1 to 20 μg / kg body weight, or about 1 to 10 μg / kg body weight.

[0132] In some embodiments, the peptide compounds or combinations of the present disclosure are provided as pharmaceutical compositions comprising a carrier. According to the present invention, a "carrier" or "pharmaceutical carrier" is a pharmaceutically acceptable solvent, suspending agent, or any other pharmacologically inert medium (carrier, vehicle) for delivering one or more peptide compounds to a subject. The carrier is typically a liquid or solid. Taking into account the intended mode of administration, when combined with the components of a given pharmaceutical composition, the pharmaceutical carrier is typically selected to provide the desired volume, consistency, etc. Typical pharmaceutical carriers include, but are not limited to, binders (e.g., pregelatinized corn starch, polyvinyl pyrrolidone or hydroxypropyl methylcellulose, etc.); fillers (e.g., lactose and other sugars, microcrystalline cellulose, pectin, gelatin, calcium sulfate, ethyl cellulose, polyacrylates or calcium hydrogen phosphate, etc.); lubricants (e.g., magnesium stearate, talc, silicon dioxide, colloidal silicon dioxide, stearic acid, metallic stearates, hydrogenated vegetable oils, corn starch, polyethylene glycol, sodium benzoate, sodium acetate, etc.); disintegrants (e.g., starch, sodium starch glycolate, etc.); and wetting agents (e.g., sodium lauryl sulfate, etc.).

[0133] The peptide compounds of the present disclosure can be administered together with one or more unit dosage forms of pharmaceutical diluents, carriers or excipients. Conventional pharmaceutical practices can be used to provide suitable formulations or compositions to administer such compositions to subjects. Although oral administration is preferred, any suitable route of administration can be used, such as intravenous, parenteral, subcutaneous, intramuscular, intracranial, intraorbital, ocular, intraventricular, intracapsular, intraspinal, intrathecal, epidural, intracisternal, intraperitoneal, intranasal, transdermal or aerosol administration. The formulation can be in the form of a liquid solution or suspension, tablet, capsule, powder for reconstruction, etc.

[0134] For example, methods for preparing formulations well known in the art are identified in Remington: The Science and Practice of Pharmacy, (19th ed.) ed. A. R. Gennaro A. R., 1995, Mack Publishing Company, Easton, PA. Formulations for parenteral administration may, for example, contain excipients, sterile water or saline, polyalkylene glycols (such as polyethylene glycol), oils of plant origin, or hydrogenated naphthalene. Biocompatible biodegradable lactide polymers, lactide / glycolide copolymers, or polyoxyethylene-polyoxypropylene copolymers may be used to control the release of these compounds. Other potentially useful parenteral delivery systems for the agonists of the present invention include ethylene vinyl acetate copolymer particles, osmotic pumps, implantable infusion systems, and liposomes. Formulations for inhalation may include excipients (e.g., lactose), or may be aqueous solutions containing, for example, polyoxyethylene-9-lauryl ether, glycocholate, and deoxycholate, or may be oily solutions for administration in the form of nasal drops or as a gel.

[0135] The peptide compounds, combinations and pharmaceutical compositions disclosed herein are useful for preventing, treating or alleviating symptoms of conditions associated with hypogonadism. The expression "prevention, treatment or alleviating of symptoms" refers to the ability of the peptide compounds, combinations and pharmaceutical compositions to limit the development, progression and / or symptoms of hypogonadism. Broadly, the prevention, treatment and / or alleviating of symptoms encompasses the reduction of symptoms associated with hypogonadism, such as, for example, infertility (due to defects or deficiencies in spermatogenesis or ovulation), aging, decreased libido, sexual dysfunction, mood changes, fatigue, decreased lean body mass, decreased bone mineral density, increased visceral fat, emaciation and / or metabolic syndrome.

[0136] The peptide compounds and combinations of the present disclosure can be administered once, twice, three times or more than once a day to achieve the desired treatment endpoint. Since the maximum effect of the peptide compounds of the present disclosure occurs between 2 and 3 hours after administration, the peptide compounds of the present disclosure can be administered once, twice, three times or even four times a day. In some embodiments, the peptide compounds of the present disclosure are administered once a day. In some embodiments, the peptide compounds of the present disclosure are administered twice a day. In some embodiments, the peptide compounds of the present disclosure are administered three times a day. In some embodiments, the peptide compounds of the present disclosure are administered four times a day. In some other embodiments, the peptide compounds or combinations can be administered in the evening or at night. Alternatively or in combination, the peptide compounds can be administered in the morning.

[0137] The present invention will be more readily understood by reference to the following examples, which are intended to illustrate the invention but not to limit its scope.

[0138] Example I - Testosterone-Inducing Peptides

[0139] Peptides. Peptides were obtained from CanPeptide (Montreal, Quebec) with a purity of >95%. All peptides were dissolved in molecular grade, sterile double distilled water to obtain 1 mM stock solutions and stored at -20°C. Peptides for oral screening were prepared from 1 mM peptide stock solutions and diluted in sterile tap water in a total volume of 1 mL. Peptides for pharmacokinetic experiments were diluted at 10 mg / ml using sterile tap water. Some of these peptides were chemically modified after synthesis. The following table summarizes the peptides that have been tested:

[0140] Table 3. Description of the peptides and peptide derivatives synthesized in this example. The symbol "d" in the amino acid nucleus refers to the D-enantiomer of the residue following the symbol used. The symbol "Ac" used throughout this specification refers to acetyl end capping (cap). The symbol "NH 2 " refers to amide end-capping. The notation "miniPEG" used throughout this specification refers to the presence of a miniPEGylated chain (NH 2 -CH 2 -CH 2 -O-CH 2 -CH 2 -O-CH 2 -CO-C 26 H 49 N 9 O 10 (647.80Da)).

[0141]

[0142]

[0143] Osmotic pumps. Model 2006 osmotic pumps, infusion 0.15 μL / hr, were purchased from Alzet (Cupertino, CA). The peptide concentration loaded into the pump was adjusted to deliver the dose indicated in each figure. The pump infusion rate varied with the production batch and was adjusted accordingly. The pump was loaded 24 hr prior to surgical implantation under sterile conditions and maintained in PBS at 37°C.

[0144] Animal handling, blood sampling, surgical procedures, and oral administration of peptides. Brown-Norway rats aged 22-28, 36-42, or 50-56 days were purchased from Charles River Laboratories (Senneville, Quebec) and aged until required. Rats were maintained on a 12L / 12D daily cycle with light on at 7AM and free access to food and water.

[0145] For all experiments, plasma samples were obtained by percutaneous jugular vein puncture and collected in EDTA KE / 1.3 tubes (Sarstedt, Numbrecht, Germany, Cat#5072511). Blood collection time points are indicated in each figure legend. Samples were centrifuged at 1300 RCF for 10 min, and plasma was stored in 2 mL Wheaton glass vials (Fisher Scientific, Hampton, NH, Cat#03337 21A, and maintained at -20°C until needed.

[0146] An osmotic infusion pump was implanted in the interscapular region of Brown-Norway rats approximately 29 days old under general anesthesia with isoflurane after iodine disinfection. The surgical wound was closed with two to three staples, which were removed one week after surgery. Carprofen (Zoetis, Parsippany, NJ) was injected subcutaneously before and two days after the surgical procedure for pain management.

[0147] For all oral experiments, dosing via gavage began at 8:30-8:50am with a 3 minute delay between rats. Animals used for pharmacokinetic studies were fasted for 4 hours prior to gavage beginning at 1pm with a 3 minute delay between rats.

[0148] Animals were handled according to protocols approved by the McGill University Animal Care and Use Committee, which included standard operating procedures for repetitive jugular vein harvesting.

[0149] Plasma steroids are measured. Testosterone levels are measured using Cayman (Ann Arbor, MI) EIA kit Cat#582701, and corticosterone levels are measured using Cayman EIA kit Cat#501320. All samples are measured in duplicate according to the manufacturer's instructions. Samples are diluted until the average control level meets the center of the standard curve. The number of animals used for each experiment is indicated in the corresponding legend.

[0150] Peptide stability, oral pharmacokinetics and quantification in plasma. For peptide stability experiments, plasma from Brown-Norway rats aged 90-140 days was collected and stored at -20°C until further use. Plasma was thawed, rotated at 12000RPM for 10 min, and kept at 4°C. 4 μM of various peptides were incubated with 100 μL plasma, which was warmed at 37°C for 10 min before use. After 1, 15, 30, 60, 90 or 120 minutes, the plasma / peptide samples were crushed with 300 μL of 100% methanol, rotated at 14000RPM for 10 minutes, and the supernatant was collected. The methanol in the supernatant was evaporated and the sample was extracted again. The final particles were dissolved in 5 μL methanol, followed by the addition of 100 μL sterile double distilled water. The extracted samples were transferred to plastic tubes and placed in the automatic loader of the mass spectrometer.

[0151] For oral pharmacokinetic experiments, jugular blood samples were collected after a single oral administration of 10 mg peptide in 1 ml of vehicle after 5, 15, 30, 45, 60, 90 and 120 minutes. A protein inhibitor cocktail (ROCHE, Total Protein Inhibitor, Cat# 11697498001) added to an EDTA collection tube was used for KVSQ and RVTQ peptides. Plasma was immediately extracted and 100 μL of plasma was triturated with 300 μL of 100% methanol, spun at 14 000 RPM for 10 min, and the supernatant was harvested. Re-extraction of the precipitant and dissolution of the precipitate were as described above.

[0152] The analysis was performed using a Thermo-Scientific ISQ Quantiva triple quadrupole mass spectrometer (QQQ) with a thermal electrospray ionization (HESI) source and a Thermo-Scientific UltiMate TM 3000UHPLC System (including UltiMate TM 3000RS autosampler). The analytical conditions were developed using reference standards in solutions containing 5 μM of the corresponding peptides. The elution was performed by isocratic elution combining a binary solvent system of (A) 0.1% formic acid (aqueous) and (B) ACN + 0.1% FA and using an Agilent Eclipse PlusTM The chromatogram was resolved using a C18 analytical column (100 mm x 2.1 mm ID, 1.8 μm particles). A 5 μL injection volume and a solvent flow rate of 150 μL / min were used. The MS / MS acquisition time was 7 minutes and the total run time was 8.3 minutes / injection. The triple quadrupole MS / MS instrument conditions were as follows: HESI source voltage was 3200 V, sheath gas 30 L / min, auxiliary gas 20 L / min, purge gas 2.0 L / min, ion transfer temperature was 350°C, evaporator temperature was 350°C, CID gas was 1.5 (mTorr), and residence time was 100 milliseconds. The Q1 resolution (FWHM) was 0.4 (unitless), and the Q3 resolution (FWHM) was 0.7 (unitless). The instrument was operated in selected reaction monitoring (SRM), positive ion detection mode at various collision energies (CE) for the following mass transitions:

[0153] Ac-RITQdS-NH2: p84(m / z 645→m / z 295, m / z 396)

[0154] RVTQ: p64(m / z 503→m / z 211);

[0155] Ac-RdITQ-NH2: p87 (m / z 558→m / z 391, m / z 267)

[0156] RVTQ: p64 (m / z 503→m / z 211; m / z 503→m / z 357 and m / z 503→m / z 256)

[0157] KVSQ: p95(m / z 461→m / z 129) and (m / z 461→m / z 234)

[0158] Statistical analysis GraphPad Prism 7.04 (GraphPad Software, La Jolla, CA) and Excel 2006 (Microsoft Corporation, Redmond, WA) were used to generate graphs, heat maps, curve fitting, and statistical analysis. ANOVA or one-tailed t-test was used to determine significant changes. The number of animals used in each experiment was recorded in each corresponding figure legend. Data are shown as mean ± standard deviation unless otherwise stated in the figure legend.

[0159] Subcutaneous infusion of RVTQ and its evolutionarily related sequences increases circulating testosterone levels

[0160] Brown-Norway rats were implanted with subcutaneous infusion pumps at approximately 29 days of age. Blood samples were collected when the rats were approximately 56 days old, and those receiving the 377 ng / kg / day peptide concentration showed a significant increase in testosterone levels ( Figure 1A ).

[0161] The UniProt database and Clustal Omega were used to search for sequence variants of the N163-166 core found in nature. S Q, R I TQ and K ITQ (underlined amino acid changes) identified as variants of nuclear sequences found primarily in fish and worms ( Figure 1B ). Sequence alignment also showed that the N-terminal amino acid flanking the N163-166 core was occupied by serine or asparagine, while the amino acid flanking the C-terminal was serine, which was conserved in all analyzed sequences. In addition, all amino acid changes identified were substituted for structurally similar amino acids.

[0162] To test whether these RVTQ-related sequences increase testosterone levels, the three variants, RVSQ, RITQ, and KITQ, were delivered using a subcutaneous infusion pump and implanted in approximately 27-day-old Brown-Norway rats. Figure 1C Samples collected at 54 days of age, where the dose of KITQ reached about 370 ng / kg / day, showed a significant increase in circulating testosterone levels. Samples collected 11 days later, at 65 days of age, when the delivered concentration was about 330 ng / kg / day, also showed a significant increase in RITQ. These results show that nuclear RVTQ allows for modification and can allow for modulation of target sites.

[0163] Oral delivery of small peptides increases testosterone levels

[0164] To determine whether RVTQ or its modified derivatives can be delivered orally, an animal model was developed in which various modifications can be effectively screened and those with orally active properties determined. Six derivatives of RVTQ were tested in Brown-Norway rats aged 60 to 123 days, which were gavaged in the morning followed by a jugular blood sample collected 3 hours later. RVTQ was used as a control, and the modifications tested included the addition of a pendant D-serine at the carboxyl terminus, amide capping, and the addition of a miniPEG or acetyl capping at the N terminus. These modifications were randomly tested with doses ranging from 160 μg / kg to 1200 μg / kg. Figure 2A RVTQdS-NH consisting of the core RVTQ (SEQ ID NO: 2) sequence plus D-serine at the carboxyl terminus and an amide cap was shown to be administered at about 580 μg / kg. 2(SEQ ID NO: 6) is a combination of doses and modifications that significantly increase testosterone levels. 2 (SEQ ID NO:6) increased testosterone levels, but the naked RVTQ (SEQ ID NO:2) sequence showed no effect at 3 h sampling time.

[0165] The optimal sampling time was then determined to confirm a significant increase in testosterone after oral administration. 2 Rats were treated in two rounds and blood samples were collected at 1 and 3 hours or 2 and 5 hours ( Figure 2B ). The pooled data showed a significant time-dependent increase in testosterone levels at 2 and 3 hours after treatment. The data also showed that treatment with RVTQdS-NH 2 Treated rats showed the best significant increase in testosterone levels after 2 hours. Based on these data, the dose and collection time point for further screening were adjusted to approximately 550 μg / kg and 2 hours, respectively.

[0166] Oral treatment with modified derivatives of RVTQ and evolutionarily related nuclear sequences increases circulating testosterone levels

[0167] To identify additional sequences / modifications with oral activity, a series of peptides consisting of modifications of the core RVTQ sequence and its evolved parent were designed. In addition, core sequences including R>K changes to the first amino acid, V>I or L changes to the second position, T>S substitutions to the third position, and Q>E to the fourth position were tested. This resulted in 10 core peptide sequences. The core sequences were further modified to include the addition of miniPEG or acetyl groups to the N-terminus, the use of D-amino acids, and capping the C-terminus with an amide group.

[0168] Brown-Norway rats aged 60 to 136 days were used and gavaged twice a week with various peptides. Each test consisted of three groups: control and two peptide combinations, for a total of 34 peptides screened. Each peptide was gavaged with a dose ranging from 550 to 620 μg / kg, followed by blood sampling at two hours ( Fig.10 The screen identified 11 nuclear / peptide modifications that increased testosterone levels ( Figure 3A -G), while the rest showed no change or non-significant decrease ( Fig.11A -J). In the final test, a significant increase in testosterone was observed, confirming that the animals still responded to the oral peptide using RITQ ( Figure 3H ). Fig. 3I Data are summarized, where peptide treatment data were normalized to their respective control testosterone levels and presented in ordered tests.

[0169] Dose-response studies and low doses

[0170] To gain further insight into the dose-response profile of the identified peptides, we tested RdVTQ and RITQ in rats aged 88 to 138 days that were gavaged daily at increasing concentrations. The results showed that RdVTQ had a significant increase ( Figure 4A ). Although RITQ showed a biphasic profile of testosterone, characterized by a significant increase at a low dose of 10 μg / kg, a non-significant decrease at about 220 μg / kg, and a significant increase at about 520 μg / kg ( Figure 4C ), but no corresponding significant changes in corticosterone levels were observed ( Figure 4B and D).

[0171] Previous dose-response experiments using RITQ indicated that the mechanism of action that induces steroidogenic effects may be subject to a biphasic response. To determine whether 11 orally active candidates showed a biphasic response at a dose of about 220 μg / kg, an experiment consisting of four groups of rats aged 82-131 days was set up. This oral assay tested the steroidogenic effects of 13 peptides administered at about 10, 220 or 420 μg / kg after two hours. The results showed that Ac-RITQdS-NH 2 , KVSQ, RdVTQ and Ac-RdITQ-NH 2 Increased mean testosterone levels, with some doses showing significant increases ( Figure 5 ). Several other peptides were identified where testosterone levels were reduced at doses of 10 μg / kg, 220 μg / kg, or 420 μg / kg (downward arrows). In addition, these results identified candidate peptides that increased testosterone levels at doses of about 420 μg / kg and at low doses of 10 μg / kg, although not significantly.

[0172] A mixture of two peptides at low doses increases plasma testosterone levels after oral administration

[0173] Previous results showed that some peptides that increased circulating testosterone levels at about 420 μg / kg had a biphasic profile characterized by suppression of testosterone levels around the 220 μg / kg dose. Results also showed that the 10 μg / kg dose increased (although not significantly) the mean testosterone levels of the six peptides ( Figure 5 ). It was hypothesized that a mixture of two peptides could result in a significant increase in testosterone levels, thereby avoiding the potential negative effects observed with some peptides. To test this hypothesis, Brown-Norway rats aged 91-134 days were treated with 15 possible permutations generated from a mixture of six candidate peptides administered at 10 μg / kg each. The results showed that five of the two-peptide combinations showed a significant increase in testosterone levels ( Fig. 6A, CF), while the rest showed no change in testosterone levels ( Fig. 12A -I). Figure 6G Peptide treatment data normalized to their respective control testosterone levels are summarized and presented in ordinal tests.

[0174] Modification of the nucleopeptide increases the half-life in serum

[0175] To determine the stability of lead candidate peptides in plasma, all peptides were incubated at 4 μM in 100 μL of plasma (except KVSQ which was incubated at 12.5 μM). The results showed that the half-life of the various peptides was as follows: Ac-RITQdS-NH 2 >RdVTQ>Ac-RdITQ-NH 2 >RITQ>RVTQ>KVSQ( Figure 7 As expected, peptide modifications with capping or introduction of D-amino acids had the longest half-lives, while peptides including K showed the fastest degradation.

[0176] Further in vivo analysis of Ac-RITQdS-NH 2 、RdVTQ、Ac-RdITQ-NH 2 , RVTQ, and KVSQ. The results showed that the peak concentrations of most peptides in the blood reached a range of 70 to 230 ng / mL within 15 to 30 minutes ( Fig. 8A -C). Bare RVTQ and KVSQ cores were not detected (data not shown).

[0177] RVTQ (N163-166) was first shown to be active in Brown-Norway rats when delivered subcutaneously. We found a significant increase in plasma testosterone levels of approximately 2-fold in Brown-Norway rats at approximately 54 days of age. We then used an evolutionary approach to investigate whether substitutions of the RVTQ core with related amino acids retained steroidogenic activity. Sequence analysis of all VDAC1 entries found in the UniProt portal identified RITQ, KITQ, and RVSQ as core variants. These substitutions were structurally related amino acids. The data showed that infusions of RITQ and KITQ significantly increased testosterone levels, indicating that changes in the protein core are feasible. Interestingly, in our first round of measurements, RITQ showed a non-significant increase, which became significant during our second round of measurements. Inhibition of peptide activity through the HPG axis is a unique "safety feature" that can prevent extended periods of supraphysiological testosterone levels.

[0178] Small molecule derivatives of the RVTQ core were designed by introducing modifications known to increase the half-life of proteins. Increased half-life was sought because rapid peptide degradation is a major challenge in using peptides as oral therapeutics. To screen for oral candidates, an animal model was developed in which Brown-Norway rats were gavaged with various RVTQ core modifications and blood was subsequently collected percutaneously from the jugular vein. The native RVTQ sequence was used as a control and the modified core derivatives were given at arbitrary doses. Blood collection was also arbitrarily set at three hours after gavage to allow for the absorption of the peptide and the time it takes for Leydig cells to increase androgen production. The data show that addition of D-serine and administration of NH 2 Capping the carboxyl terminus resulted in a significant increase in testosterone levels. This finding was confirmed in two subsequent experiments characterizing the optimal sampling time point, which was 2 hours after gavage with approximately 580 μg / kg of RVTQdS-NH 2 Stable. The data show that RVTQdS-NH 2 is rapidly absorbed, and the peptide reaches the Leydig cells in sufficient quantities to effectively increase steroidogenesis. Furthermore, these findings are substantial because the increase in steroid levels during the 2 to 3 hour window may escape HPG suppression and allow for multiple dosing. The kinetics of this peptide also facilitate the development of dose simulations that mimic the physiological peaks in testosterone production found at specific times of the day. Furthermore, the oral route provides the best compliance and allows for dosing flexibility.

[0179] The various peptides were then tested using Brown-Norway rats, and blood collection was established 2 hours after gavage with peptide doses ranging from an average of 530 to 620 μg / kg. This screen included molecules with amino acid modifications to the RVTQ core where R was substituted with K, V was substituted with I or L, T was substituted with S, and Q was substituted with E. These amino acid modifications generated 10 core sequences, which when combined with D-amino acids to cap the peptide ends, and using miniPEG, generated 34 candidate small molecules. This screen identified 11 small molecules that significantly increased testosterone levels. To confirm that the effect was still present at the end of our screen, we treated the rats with RITQ at the end of the experiment, identified the first molecule, and observed a significant increase in testosterone. Again, this screen was initiated when the rats were approximately 60 days old; however, the data showed that rats approximately 80-90 days old were a better starting point because younger rats exhibited higher testosterone levels, which could mask the peptide effect. Unexpectedly, the range of amino acid changes and protein modifications that increased testosterone levels were noted, including unmodified core R I TQ, K V S Q, R IS Q. KI TQ, RV S Q. andS RVTQ (underlined letters indicate amino acid changes relative to the original RVTQ (SEQ ID NO: 2) core). In general, modifications to V were most permissive, with the exception of L where the additional carbon could cause steric hindrance. The data also suggest that Q is essential, as any substitution abolished peptide activity. This is consistent with the analysis guided by the initial tuning, which showed that Q was conserved across all sequences treated ( Figure 1B ). Several other peptides were identified that showed a non-significant trend to increase testosterone levels, and it is hypothesized that some of these peptides may be active if the dose is adjusted. In addition, several other peptides were identified that had a non-significant trend to decrease testosterone levels, which may indicate a dose-dependent inhibitory state.

[0180] Dose-response experiments were performed using RdVTQ, which showed the best performance, and RITQ, which showed the second best statistical significance. The results showed that RdVTQ increased the average testosterone levels at all tested doses, reaching significance at doses higher than 400 μg / kg. RITQ showed a biphasic response, in which testosterone levels increased significantly at 10 and 510 μg / kg, and did not significantly decrease at about 210 μg / kg. Interestingly, RdVTQ also showed an increasing trend at a low dose of about 10 μg / kg, followed by a slight decrease in testosterone levels. It is important to note that testosterone levels were collected 2 hours after gavage, and when testosterone levels increased, the integral of the area under the curve for each rat during 3 hours may show a significant increase. In short, these results show that the intracellular mechanism triggered by the administration of peptides can have a self-regulatory state, resulting in the biphasic effect shown. These dose-response experiments also show that the small molecules tested do not affect adrenal steroidogenesis because there is no significant change in corticosterone levels.

[0181] We then proceeded to identify peptides that robustly increased testosterone levels at doses that confirmed the biphasic effect of RITQ. Testing conditions were improved by using rats older than 80 days, collecting blood samples 2 hours after gavage, and dosing at approximately 10, 220, and 420 μg of peptide / day. Eleven (11) molecules that significantly increased testosterone were tested, including Ac-RITQdS-NH 2 , i.e., the first peptide that was excluded from the inclusion criteria but showed a trend toward increased testosterone levels. This screen identified Ac-RITQdS-NH 2 , KVSQ, RdVTQ and Ac-RdITQ-NH 2The peptides were identified as those that consistently increased testosterone levels in the doses tested. Several other peptides were identified that showed a reduction in steroid levels at about 220 μg / kg, including RITQ, consistent with the first observation of a biphasic effect. In summary, the third screen identified four peptides with robust spectra, and in addition, the data showed an increase (although not significant) in Ac-RITQdS-NH at a dose of about 10 μg / kg. 2 、RdVTQ、Ac-RdITQ-NH 2 , RITQ, KVSQ, and Ac-KdITQ-NH 2 Average testosterone levels.

[0182] We then sought to improve the efficacy of the selected peptides to reduce the dosage used while increasing the safety margin. A combination of two of the six peptides (due to their modifications, which may have different pharmacokinetic profiles) was tested to determine if they could work together to exhibit a significant effect. The peptide Ac-RITQdS-NH was specifically selected 2 、KVSQ、RdVTQ、Ac-RdITQ-NH 2 , RITQ, and the original RVTQ core tetrapeptide. The permutations of these 6 small molecules consisting of 3 natural cores and 3 modified cores resulted in 15 possible 2-peptide combinations tested. The testing conditions were set as follows: Brown-Norway rats were >90 days old, blood was collected 2 hours after gavage, and 10 μg / kg of each of the 2 peptides was used for testing. The results confirmed 5 combinations that significantly increased testosterone levels. This finding is significant because it is a 55-fold increase in the dose compared to the first and second screenings that we set at approximately 550 μg / kg. The very low doses used in the oral 2-peptide mixture may avoid side effects, be quickly cleared, facilitate repeated dosing, and reduce manufacturing costs.

[0183] The pharmacokinetic profiles of the six lead peptides were characterized in plasma and in vivo. The data showed that Ac-RITQdS-NH 2 is the most stable peptide, followed by RdVTQ and Ac-RdITQ-NH 2 . The unmodified core tetrapeptides RITQ and RVTQ showed rapid time-dependent degradation. The data for KVSQ (the input peptide was 4 times larger than all other peptides tested) indicated that the presence of lysine significantly increased peptide degradation in the in vitro assay. The pharmacokinetic profiles of these drugs were characterized in vivo. The data showed that most peptides reached peak concentrations between 15 and 30 minutes after gavage, with Ac-RITQdS-NH 2Absorption is fastest in about 15 minutes. These data (showing rapid loss of the peptide) are consistent with the finding that testosterone levels reach peak induction levels at 2 hours. Moreover, the rapid degradation of KVSQ in both in vitro and in vivo assays is unexpected because this tetrapeptide appears to consistently activate steroidogenesis. The biological activity and rapid degradation of KVSQ suggest that only very few molecules are needed to activate the molecular target and elicit an effect.

[0184] Example II - TV159-172 Derivatives that Regulate Testosterone Levels

[0185] Peptides. Peptides were obtained from CanPeptide (Montreal, Quebec) with a purity of >95%. All peptides were dissolved in molecular grade double distilled water to obtain 1 mM stock solutions and stored at -20°C.

[0186] Animal Handling, Serum, Plasma and Intratesticular Sample Collection Brown-Norway rats were purchased from Charles River at 36-42 days of age and aged until approximately 60 days of age. Rats were maintained on a 12L / 12D daily cycle with lights on at 7AM and ad libitum access to food and water.

[0187] Jugular vein samples were collected by percutaneous puncture and collected in EDTA-coated tubes (plasma) and centrifuged at 1,300 RCF for 10 minutes and stored in 2 mL Wheaton glass vials (Fisher Scientific, Hampton, NH, Cat#03 337 21A) and stored at -20° C. Animals were handled according to protocols approved by the McGill University Animal Care and Use Committee, which included standard operating procedures for the amount and frequency of blood sample collection.

[0188] Hormone Measurements. Testosterone levels were measured using Cayman (Ann Arbor, MI) EIA Kit Cat# 582701, and corticosterone levels were measured using Cayman EIA Kit Cat# 501320. Plasma fluid samples were diluted until the control matched the middle of the standard curve used.

[0189] Osmotic pumps. Alzet 2006 osmotic pumps were purchased from Alzet (Cupertino, CA) with an infusion rate of 0.15 μL / hr. The peptide concentration loaded into the pumps accounted for body weight gain at the time of collection and was adjusted to deliver the dose indicated in each figure. Pump infusion rates varied with production batches and were adjusted accordingly. Pumps were loaded under sterile conditions 24 hours prior to surgical implantation and maintained in PBS at 37°C until use.

[0190] Surgical Procedure. After disinfection with iodine, the infusion pump was implanted in the interscapular region under general anesthesia. The surgical wound was closed with staples, which were removed one week after surgery. Carprofen (Zoetis, Parsippany, NJ) was injected subcutaneously for pain management before and two days after the surgical procedure.

[0191] Statistical analysis GraphPad Prism 7.03 (GraphPad Software, La Jolla, CA) and Excel 2006 (Microsoft Corporation, Redmond, WA) were used to generate graphs, linear regressions, and statistical analyses. Analysis of variance or one-tailed t-tests were used to determine significant changes. The number of animals used in each experiment is given in each corresponding figure legend. Data are shown as mean ± standard deviation unless otherwise stated in the figure legend.

[0192] Shorter derivatives of TV159-172 are biologically active in inducing steroidogenesis

[0193] The various experimental groups of Brown-Norway rats were implanted with subcutaneous pumps delivering increasing doses of TV-peptides for 42 days. Single amino acid deletions were performed stepwise from the N-terminal and C-terminal ends until the shortest peptides TV163-167 and TV162-166 were reached, which comprised 5 amino acids from the original 14 amino acid VDAC1 derived sequences contained in TV159-172. Testosterone measurements in the plasma collected 1 week after implantation were shown to significantly increase testosterone levels in TV156-172, TV160-169, TV161-170, TV162-169, TV161-167, TV162-166 ( Figures 13A-13I Pooling of all control data and doses from all rats treated with TV-peptides (independent of dose) showed increased dispersion in all TV-peptides ( Fig.13J ). The pooled results showed that while the highest control reached 2.6 times the control mean, rats receiving TV-peptide often had levels higher than 2.6 times, and in some cases, higher than 4 times the control mean.

[0194] Plasma corticosterone levels collected on day 40 of infusion were significantly increased by exposure to TV160-171, TV160-170, and TV162-169, but significantly decreased in response to TV161-167 treatment ( Figures 14A-14I ).

[0195] Tetrapeptide N163-166 increases testosterone levels

[0196] Deletion experiments showed that the shorter peptides TV163-167 and TV162-166 had steroid activity. Fig.15A It was shown that TV163-167 and TV162-166 overlapped at amino acids 163-166 of their RVTQ sequence corresponding to a small molecule weight of 503 daltons. Brown-Norway rats were subcutaneously injected with increasing concentrations of N163-166, where N represents the "naked" and TAT-free nature of this molecule. Testosterone and corticosterone levels were measured 7 and 40 days after injection, respectively. The results obtained showed that N163-166 significantly increased testosterone levels ( Fig. 15B ). Corticosterone levels were not affected by treatments that induced testosterone formation and were reduced at a lower dose of 23 ng / kg / day of the tetrapeptide N163-166 ( Fig. 15C ).

[0197] Based on the VDAC1 sequence, the confirmation of the minimum biologically active sequence is sought in the confirmed TV peptide. The biologically active core retaining steroid activity is found in the TV159-172 sequence using Brown-Norway rats. The progressive deletion of the TV159-172 sequence shows that the steroidogenic activity can be maintained with a short 5-amino acid sequence. The testosterone level data induced by different peptides are summarized (independent of the dosage used) to confirm the overall activity of IV deletion. The summary analysis of the control rats composed of 60 samples obtains a rat with a summary control average level of 2.6 times the control rat with the highest testosterone level. In contrast, all peptides tested have rats with several testosterone levels exceeding the highest control rats, some of which are up to 5 times more than the summary control average. Although the results of the summary are not meant for statistical analysis, they show that rat separation (i.e. TV160-169 and TV162-169) is a low-level and high-level responder, indicating that some rats reach a testosterone level that triggers the regulation of the HPG axis. Testosterone levels have circadian rhythms and seasonal variations, and the impact of these factors on the effect of TV peptides remains to be studied. In summary, the results show that shorter sequences derived from TV159-172 are active in stimulating testosterone biosynthesis in Leydig cells.

[0198] The measurement of plasma cortisol level in the same Brown-Norway rats processed with various peptides shows peptide-specific activity.Longer TV160-171, TV160-170 and TV162-169 peptides promote the significant increase of plasma corticosterone levels, and the shortest TV163-167 and TV162-166 do not show significant changes.Some corticosterone levels (as seen in TV160-170) are higher than those seen in testosterone measurement, and up to 15 times of control mean value.These results show that some rats show very high corticosterone levels, because different from testosterone, corticosterone is not subject to strict regulation.It is important to note that the percutaneous collection of blood samples is not the best method for evaluating adrenal effect.This is because the collection process affects the stress level in these rats, and therefore affects the release of corticosterone.Nevertheless, the effect observed in TV160-171 and TV160-170 shows a dose-response effect. In conclusion, the data obtained suggest that the length of TV159-172-derived sequences has differential effects on testicular and adrenal steroid production, with longer sequences favoring corticosterone production, whereas shorter sequences target testosterone biosynthesis in Leydig cells.

[0199] Since the shorter TV163-167 and TV162-166 peptides showed androgenic activity and reduced corticosterone induction, we wondered whether the RVTQ sequence (which corresponds to the shared amino acids 163-166) had steroidogenic effects. Results obtained using the same subcutaneous infusion model showed that N163-166 was biologically active. A significant increase in plasma testosterone was observed, but no change in corticosterone levels at the highest dose tested, and a significant reduction in corticosterone levels at the lowest dose tested.

[0200] In a word, N163-166 increases testosterone production without affecting adrenal steroid production. As indicated by testosterone production, subcutaneous infusion of TV- and N (naked)-peptides arrives at Leydig cells, although these peptides span various compartments, with enough levels. Although subcutaneous infusion is feasible for therapeutic purposes, here, it is used as a means to help develop original TV159-172 peptides. The discovery that core N163-166 has biological activity will significantly help develop this molecule as a therapeutic agent and the method for selectively increasing the treatment delivery of androgen levels.

[0201] While the present invention has been described in conjunction with specific embodiments thereof, it will be understood that the scope of the claims should not be limited by the preferred embodiments set forth in the examples, but should be given the broadest interpretation consistent with the description as a whole.

[0202] References

[0203] Aghazadeh,Y.,et al.,Induction of Androgen Formation in the Male byaTAT-VDAC1 Fusion Peptide Blocking 14-3-3varepsilon Protein Adaptor andMitochondrial VDAC1 Interactions. Mol Ther, 2014.22(10):p.1779-91. Sequence Listing <110> The Royal Institute for the Advancement of Learning / McGill University LEARNING / MCGILL UNIVERSITY) <120> Testosterone-inducing peptide compounds and related combinations <130> 56080224-2PCT <150> 62 / 756767 <151> 2018-11-07 <160> 34 <170> PatentIn version 3.5 <210> 1 <211> 4 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptides <400> 1 Lys Val Ser Gln 1 <210> 2 <211> 4 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptides <400> 2 Arg Val Thr Gln 1 <210> 3 <211> 4 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptides <220> <221> Variants <222> (2)..(2) <223> Xaa is D-valine <400> 3 Arg Xaa Thr Gln 1 <210> 4 <211> 4 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptides <220> <221> Variants <222> (3) <223> Xaa is D-threonine <400> 4 Arg Val Xaa Gln 1 <210> 5 <211> 4 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptides <400> 5 Arg Val Ser Gln 1 <210> 6 <211> 4 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptides <220> <221> Variants <222> (2)..(2) <223> Xaa is D-valine <400> 6 Arg Xaa Ser Gln 1 <210> 7 <211> 4 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptides <220> <221> Variants <222> (3) <223> Xaa is D-serine <400> 7 Arg Val Xaa Gln 1 <210> 8 <211> 4 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptides <400> 8 Arg Ile Thr Gln 1 <210> 9 <211> 4 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptides <220> <221> Variants <222> (2)..(2) <223> Xaa is D-isoleucine <400> 9 Arg Xaa Thr Gln 1 <210> 10 <211> 4 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptides <400> 10 Lys Ile Thr Gln 1 <210> 11 <211> 4 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptides <220> <221> Variants <222> (2)..(2) <223> Xaa is D-isoleucine <400> 11 Lys Xaa Thr Gln 1 <210> 12 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptides <400> 12 Arg Val Thr Gln Ser 1 5 <210> 13 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptides <220> <221> Variants <222> (5) <223> Xaa is D-serine <400> 13 Arg Val Thr Gln Xaa 1 5 <210> 14 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptides <400> 14 Arg Ile Thr Gln Ser 1 5 <210> 15 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptides <220> <221> Variants <222> (5) <223> Xaa is D-serine <400> 15 Arg Ile Thr Gln Xaa 1 5 <210> 16 <211> 4 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptides <400> 16 Arg Ile Ser Gln 1 <210> 17 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptides <400> 17 Ser Arg Val Thr Gln 1 5 <210> 18 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptides <400> 18 Ser Arg Val Thr Gln 1 5 <210> 19 <211> 4 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptides <400> 19 Lys Ile Ser Gln 1 <210> 20 <211> 4 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptides <220> <221> Variants <222> (2)..(2) <223> Xaa is D-isoleucine <400> 20 Lys Xaa Ser Gln 1 <210> twenty one <211> 4 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptides <400> twenty one Arg Val Thr Glu 1 <210> twenty two <211> 4 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptides <220> <221> Variants <222> (2)..(2) <223> Xaa is D-valine <220> <221> Variants <222> (3) <223> Xaa is D-threonine <400> twenty two Arg Xaa Xaa Gln 1 <210> twenty three <211> 6 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptides <220> <221> Variants <222> (1) <223> Xaa is D-serine <220> <221> Variants <222> (6) <223> Xaa is D-serine <400> twenty three Xaa Arg Val Thr Gln Xaa 1 5 <210> twenty four <211> 4 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptides <220> <221> Variants <222> (1) <223> Xaa is lysine or arginine <220> <221> Variants <222> (2)..(2) <223> Xaa is valine, isoleucine, or leucine <220> <221> Variants <222> (3) <223> Xaa is serine or threonine <220> <221> Variants <222> (4) <223> Xaa is glutamine or glutamic acid <400> twenty four Xaa Xaa Xaa Xaa 1 <210> 25 <211> 6 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptides <220> <221> Variants <222> (1) <223> Xaa may be present or absent, and when present may be serine <220> <221> Variants <222> (2)..(2) <223> Xaa is lysine or arginine <220> <221> Variants <222> (3) <223> Xaa is valine, isoleucine, or leucine <220> <221> Variants <222> (4) <223> Xaa is serine or threonine <220> <221> Variants <222> (5) <223> Xaa is glutamine or glutamic acid <220> <221> Variants <222> (6) <223> Xaa may be present or absent, and when present may be serine <400> 25 Xaa Xaa Xaa Xaa Xaa Xaa 1 5 <210> 26 <211> 8 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptides <220> <221> Variants <222> (1) <223> Xaa may be present or absent, and when present may be any amino acid <220> <221> Variants <222> (2)..(2) <223> Xaa may be present or absent, and when present may be serine <220> <221> Variants <222> (3) <223> Xaa is lysine or arginine <220> <221> Variants <222> (4) <223> Xaa is valine, isoleucine, or leucine <220> <221> Variants <222> (5) <223> Xaa is serine or threonine <220> <221> Variants <222> (6) <223> Xaa is glutamine or glutamic acid <220> <221> Variants <222> (7) <223> Xaa may be present or absent, and when present may be serine <220> <221> Variants <222> (8) <223> Xaa may be present or absent, and when present may be any amino acid <400> 26 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 1 5 <210> 27 <211> 10 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptides <220> <221> Variants <222> (1) <223> Xaa may be present or absent, and when present may be any amino acid <220> <221> Variants <222> (2)..(2) <223> Xaa may be present or absent, and when present may be serine <220> <221> Variants <222> (3) <223> Xaa may be present or absent, and when present may be proline <220> <221> Variants <222> (4) <223> Xaa may be present or absent, and when present may be serine <220> <221> Variants <222> (5) <223> Xaa is lysine or arginine <220> <221> Variants <222> (6) <223> Xaa is valine, isoleucine, or leucine <220> <221> Variants <222> (7) <223> Xaa is serine or threonine <220> <221> Variants <222> (8) <223> Xaa is glutamine or glutamic acid <220> <221> Variants <222> (9)..(9) <223> Xaa may be present or absent, and when present may be serine <220> <221> Variants <222> (10)..(10) <223> Xaa may be present or absent, and when present may be any amino acid <400> 27 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 1 5 10 <210> 28 <211> 11 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptides <220> <221> Variants <222> (1) <223> Xaa may be present or absent, and when present may be any amino acid <220> <221> Variants <222> (2)..(2) <223> Xaa may be present or absent, and when present may be serine <220> <221> Variants <222> (3) <223> Xaa is lysine or arginine <220> <221> Variants <222> (4) <223> Xaa is valine, isoleucine, or leucine <220> <221> Variants <222> (5) <223> Xaa is serine or threonine <220> <221> Variants <222> (6) <223> Xaa is glutamine or glutamic acid <220> <221> Variants <222> (7) <223> Xaa may be present or absent, and when present may be serine <220> <221> Variants <222> (8) <223> Xaa may be present or absent, and when present may be arginine <220> <221> Variants <222> (9)..(9) <223> Xaa may be present or absent, and when present may be phenylalanine <220> <221> Variants <222> (10)..(10) <223> Xaa may be present or absent, and when present may be alanine <220> <221> Variants <222> (11)..(11) <223> Xaa may be present or absent, and when present may be any amino acid <400> 28 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 1 5 10 <210> 29 <211> 13 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptides <220> <221> Variants <222> (1) <223> Xaa may be present or absent, and when present may be any amino acid <220> <221> Variants <222> (2)..(2) <223> Xaa may be present or absent, and when present may be serine <220> <221> Variants <222> (3) <223> Xaa may be present or absent, and when present may be proline <220> <221> Variants <222> (4) <223> Xaa may be present or absent, and when present may be serine <220> <221> Variants <222> (5) <223> Xaa is lysine or arginine <220> <221> Variants <222> (6) <223> Xaa is valine, isoleucine, or leucine <220> <221> Variants <222> (7) <223> Xaa is serine or threonine <220> <221> Variants <222> (8) <223> Xaa is glutamine or glutamic acid <220> <221> Variants <222> (9)..(9) <223> Xaa may be present or absent, and when present may be serine <220> <221> Variants <222> (10)..(10) <223> Xaa may be present or absent, and when present may be arginine <220> <221> Variants <222> (11)..(11) <223> Xaa may be present or absent, and when present may be phenylalanine <220> <221> Variants <222> (12)..(12) <223> Xaa may be present or absent, and when present may be alanine <220> <221> Variants <222> (13) <223> Xaa may be present or absent, and when present may be any amino acid <400> 29 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 1 5 10 <210> 30 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptides <400> 30 Lys Ser Arg Val Thr Gln Ser 1 5 <210> 31 <211> 8 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptides <400> 31 Ser Arg Val Thr Gln Ser Asn Phe 1 5 <210> 32 <211> 10 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptides <400> 32 Lys Ser Arg Val Thr Gln Ser Asn Phe Ala 1 5 10 <210> 33 <211> 10 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptides <400> 33 Ser Lys Ser Arg Val Thr Gln Ser Asn Phe 1 5 10 <210> 34 <211> 11 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptides <400> 34 Ser Lys Ser Arg Val Thr Gln Ser Asn Phe Ala 1 5 10

Claims

1. Choose from RVTQ, RISQ, RVSQ, RITQ, RdVTQ, Ac-KITQ-NH 2 、Ac-RdITQ-NH 2 、KVSQ、RdITQ、Ac-RISQ-NH 2 and KITQ-NH 2 Use of the peptides in the group consisting of for preparing a medicament for promoting endogenous steroid production in cells.

2. The use according to claim 1, in, The steroid is testosterone.

3. The use according to claim 1 or 2, in, The cell is in vivo.

4. The use according to claim 3, further comprising administering a therapeutically effective amount of the peptide or the drug to a subject in need thereof and comprising the cell.

5. The use according to claim 4, in, The subject is a mammal.

6. The use according to claim 4, in, The subject is male.

7. The use according to claim 1 or 2, in, The cells are from the testes.

8. The use according to claim 7, in, The cells are Leydig cells.

9. Use according to claim 1 or 2, for preventing, treating and / or alleviating the symptoms of a condition associated with hypogonadism.

10. The use according to claim 9, in, The condition associated with hypogonadism is at least one of infertility, aging, decreased libido, sexual dysfunction, mood changes, fatigue, decreased lean body mass, decreased bone mineral density, increased visceral fat, wasting, or metabolic syndrome.

11. The use according to claim 9, in, The hypogonadism is primary hypogonadism, secondary hypogonadism, tertiary hypogonadism or acquired hypogonadism.

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