MXPA00008537A - NON-PEPTIDE GnRH AGENTS - Google Patents

Abstract

Non-peptide GnRH agents capable of inhibiting the effect of gonadotropin-releasing hormone are of general formula (I), where X1, X2, Y, and Z are defined variables. Such compounds and their pharmaceutically acceptable salts, multimers, prodrugs, and active metabolites are suitable for treating mammalian reproductive disorders and steroid hormone-dependent tumors as well as for regulating fertility, where suppression of gonadotropin release is indicated. Methods for synthesizing the compounds and intermediates useful in their preparation are also described.

Description

NON-PEPTIDE HUMAN GONADOTROPIN AGENTS.

TECHNICAL FIELD AND INDUSTRIAL ID.M3 APPLICATION OF THE INVENTION This invention is generally related to compounds that affect the action of the hormone. that releases gonadotropin human (GnRH). More particularly, it is related to non-peptidic GnRH antagonists or agonists and their preparation. These non-peptidic GnRH agents have physical properties. chemical and biological advantages, and are useful drugs for diseases or conditions mediated by modulation of the pituitary-gonad axis. The compounds of the invention avoid the problems of degradation and biodistribution of peptide agents.

BACKGROUND OF THE INVENTION The release of a hormone (a biochemical substance that is produced by a specific cell or tissue and that causes a change or activity in a cell or tissue located elsewhere in the body) by the anterior lobe of the gland pituitary (which is located at the base of the brain and secretes hormones related to sexual growth and development) normally requires the prior release of another class of hormones produced by the REF .: 122464 hypothalamus (a structure in the lower part of the brain that connects to and controls the pituitary gland). One of the actions of hypothalamic hormones as a factor that activates the release of gonadotropin hormones, particularly LH (luteinization hormone, which is the pituitary hormone that causes the testes in men and ovaries in women to elaborate the sex hormones) and FSH (follicle stimulation hormone, which is the pituitary hormone that stimulates follicle growth in women and the formation of sperm in men). This hormone is referred to herein as "GnRH" (gonadotropin-releasing hormone) and / or "LH-RH" (hormone that releases luteinization hormone). GnRH is a decapeptide hormone produced by the arcuate nuclei of the hypothalamus (an arcuate nucleus is any of the cell masses in the thalamus, hypothalamus, or medulla oblongata) that controls the production and release of the gonadotropin pituitary gland (the hormones include FSH and LH that are produced by the pituitary gland that controls reproductive function). The GnRH (LH-RH) can be represented by the pyro-Glu-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NH2 sequence or, in the code designation - single-letter, pyro- EHWSYGLRPG-NH2. GnRH acts on high affinity pituitary receptors to stimulate the production and release of LH and FSH. The pituitary response to GnRH varies greatly throughout life. GnRH and gonadotropins first appear in the fetus at approximately ten weeks of gestation. The sensitivity to rejection of GnRH, after a brief increase during the first three months after birth, until the beginning of puberty. Before puberty, the response of FSH to GnRH is greater than that of LH. Once puberty starts, sensitivity to GnRH increases, and the secretion of pulsatile LH arises. After puberty and throughout the reproductive years, pulsations occur throughout the day, with sensitivity to LH greater than that of FSH. After menopause, FSH and LH levels rise, and in postmenopause, FSH levels are higher than LH. The release of pulsatile GnRH results in the release of pulsatile LH and FSH. However, sustained infusion of GnRH and its analogues result in inhibition of the release of LH and FSH. This phenomenon has been used in the successful treatment of precocious gonadotropin puberty mediated by the sustained administration of LH-RH or its analogues. Reciprocally, in people with GnRH deficiency, the pulsatile administration of LH-RH can restore a normal menstrual cycle or normal sperm and the production of testosterone. GnRH agonists, which are compounds that stimulate the pituitary gland to release or modulate FSH and LH, has been the mode of choice for treating sex steroid-dependent pathophysiologies, due to the limited number of appropriate antagonists available for clinical evaluation. GnRH antagonists, which are compounds that suppress the pituitary gland of the release of FSH and LH, however, are now considered. GnRH antagonists may be useful for suppressing gonadotropin secretions and preventing ovulation or in female mammals. GnRH antagonists have been investigated for contraception and to regulate periods of conception, as well as to treat infertility, to control the induction of ovulation in women with chronic anovulation, and for in vitro fertilization. GnRH antagonists are also useful for the treatment of precocious puberty, endometriosis (including endometriosis with pain), acne, amenorrhea (eg secondary amenorrhea), uterine myoma, ovarian diseases and mammary cysts (including polyvalent ovarian disease). ), and breast and gynecological cancers. GnRH antagonists may also be useful in the symptomatic release of premenstrual syndrome (PMS). They can also be used to treat ovarian hyperandrogenism and hirsutism. Antagonists have also been found useful in regulating the secretion of gonadotropins in male mammals and can be used for the arrest of spermatogenesis, for example, as male contraceptives for the treatment of male sex offenders, and for the treatment of prostatic hypertrophy. . More specifically, GnRH antagonists can be used to treat steroid-dependent tumors, such as prosthetic and mammary rumors, and to control the ovulation period for in vitro fertilization. GnRH antagonists can also be used to treat patients who have diseases, such as AIDS, where stimulation of the thymus to produce T cells would be beneficial. All uses are related to the ability of the GnRH antagonist to block GnRH activity.Previously, the available GnRH antagonists have been mainly peptide analogs of GnRH. See, for example, International Publication No. WO 93/03058. Peptide antagonists of peptide hormones are often really potent; however, the use of peptide antagonists is usually associated with problems because the peptides are degraded by physiological enzymes and are often poorly distributed within the organism to be treated. Therefore, effectiveness has been limited as medicines. Accordingly, there is currently a need for non-peptide antagonists of the GnRH peptide hormone.

BRIEF DESCRIPTION OF THE INVENTION An object of the invention is therefore to provide non-peptidic compounds that are GnRH- (agonist or antagonist) agents that bind to GnRH receptors and their modulating activity, especially those that are potent GnRH antagonists. Another object of the invention is to provide effective therapies to individuals in need of therapeutic regulation of GnRH and to provide methods for treating diseases and conditions mediated by the regulation of GnRH. - these objectives have been achieved by the non-peptidic GnRH compounds of the invention, which are useful as pharmaceuticals for indications mediated by the regulation of GnRH. The compounds of the invention are pharmaceutically advantageous over peptide compounds since they provide better biodistribution and tolerance to degradation by physiological enzymes. The invention further provides methods for synthesizing the compounds as well as intermediary compounds useful for making the compounds. The GnRH agents of the invention are of the general Formula I: (I) wherein: Z is a group selected from alkyl, alkenyl, alkynyl, cycloalkyl, heterocycle, ary, heteroaryl, CH20R, and C (O) OR, substituted or unsubstituted, wherein R is alkyl, alkenyl, alkynyl, cycloalkyl, aryl, or heteroaryl, substituted or unsubstituted and where the total number of carbon atoms present in Z, optionally does not include substituents, ranging from 1 to 12; Y is a lipophilic group selected from alkyl, alkenyl, alkynyl, cycloalkyl, aryl, and heteroaryl, substituted or unsubstituted, where the total number of carbon atoms present in Y, optionally does not include substituents, ranging from 6 to 20; Xi is a structural unit, or in space, used to join the functional units CH2NC (O), X2, Y, and Z in the three-dimensional space, which is selected from alkyl, alkenyl, alkynyl, cycloalkyl, aryl, and substituted heteroaryl and unsubstituted such that the count of atoms in the chain portion of the unit linking the central nitrogen (the N atom in Formula I is referred to as "central nitrogen" to avoid confusion with any other nitrogen-produced substituent) for X2 goes from 3 to 8, preferably from 4 to 6; and X2 is a basic group having a pKa greater than about 8 which is preferably selected from guanidinyl, amidinyl, acylamidinyl, azetidinyl, and unsubstituted and substituted amino. Additionally for compounds of Formula I, the GnRH agents of the invention include pharmaceutically acceptable salts, multimeric forms, prodrugs, and active metabolites of the compounds of Formula I. Non-peptidic agents are pharmaceutically advantageous over peptidic agents which provide better biodistribution and tolerance to degradation by physiological enzymes. The invention also relates to pharmaceutical compositions comprising a therapeutically effective amount of a GnRH agent of the invention in combination with a pharmaceutically acceptable carrier or diluent.In addition, the invention relates to methods for regulating gonadotropin secretion in mammals, which comprise therapeutically administering effective amounts of GnRH agents of the invention The invention is further related to the processes for synthesizing the compounds as well as for intermediary compounds useful for making the compounds The intermediate compounds useful for making the compounds of the formula I are those encompassed by the following formulas II, III, and IV where: j e s 1 or 2; k e s 1, 2, 3, 4 or 5; Re is H or a substituted or unsubstituted lower alkyl; R9 is H or a substituted or unsubstituted lower alkyl, CN, N02 or CO? R1; R10 is H or lower alkyl, CH2OR1, (CH2) POR1, COcR1, or (CH2) PC (0) R2, where p is an integer from 1 to 6, and R2 is H, OR1, SR1, N (R1 ) 2, or 0 (^) 3 substituted or unsubstituted; R11 is H or lower alkyl, CH20-phenyl, CH20-benzyl, phenyl, or benzyl, substituted or unsubstituted; or any two of R8, R9, R10, and R11 (R8 and R9, or R8 and R11 or R8 and R10, or R9 and R10, or R9 and R11, or R10 and R11) which together form a heterocycle with 5- or 6-members; and wherein each R1 is independently selected from H and lower alkyl, O-lower alkyl, and S-substituted or unsubstituted lower alkyl. Other features, objects, and advantages of the invention will become clear from the following detailed description of the invention and its preferred embodiments.

BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 is a plane of a displacement curve derived from the measurements of the amount of radiolabelled or radiolabeled GnRH bound to the membranes that produce the GnRH receptors against the concentration of unlabelled or unlabelled GnRH. Figure 2 is a plane of percent acidification in human GnRH receptor expression cells in response to the increased concentration of GnRH in the absence and presence of a GnRH antagonist.

DETAILED DESCRIPTION AND PREFERRED MODALITIES OF THE INVENTION As-'en is of GnRH Some of the compounds of the invention contain one or more centers of asymmetry, and can thus give rise to enantiomers, diastereoisomers, and other stereoisomeric forms. The invention mainly includes all possible stereoisomers as well as their racemic and optically pure forms. When the compounds described herein contain olefinic double bonds, it is thought that they encompass both geometric E Z isomers. The chemical formulas referred to herein may show the phenomenon of tautomerism. As the structural formula shown in this specification only describes one of the possible tautomeric forms, it should be understood however that the invention encompasses all tautomeric forms. The term "alkyl" refers to straight-chain and branched-chain groups having one to twelve carbon atoms. Exemplary alkyl groups include methyl (Me), ethyl, n-propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl (tBu), penryl, isopentyl, tert-pentyl, hexyl, isohexyl, and the like. The term "lower alkyl" refers to an alkyl having 1 to 8 carbon atoms (an alkyl of 1 to 8 carbon atoms). Substituted alkyls include fluoromethyl, trifluoromethyl, 2-fluoroethyl, 3-fluoropropyl, hydroxymethyl, 2-hydroxyethyl, 3-hydroxypropyl, and the like. The term "alkenyl" refers to straight-chain and branched alkenyl groups having from 2 to 12 carbon atoms. Exemplary alkenyl groups include prop-2-enyl, but-2-enyl, but-3-enyl, 2-methyl-prop-2-enyl, hex-2-enyl, and the like. The term "alkynyl" refers to straight-chain and branched alkynyl groups having from 2 to 12 carbon atoms. Exemplary alkynyl include prop-2-ynyl, 3-methylpent-4-ynyl, hex-2-ynyl, and the like. The term "carbocycle" refers to a monocyclic or polycyclic carbon ring structure (without the heteroatoms) having from 3 to 7 carbon atoms in each ring, which may be partially saturated, or unsaturated. Exemplary carbocycles include cycloalkyls and aryls.

The term "heterocycle" refers to a monocyclic or polycyclic ring structure with one or more heteroatoms selected from N, O, and S, and having from 3 to 7 atoms (more carbon atoms than any heteroatom (s)) in Each ring, which can be saturated, partially saturated, or unsaturated. Exemplary hexocycles include tetrahydrofuranyl, tetrahydropyranyl, azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, and the like. The term "cycloalkyls" as used herein refers to saturated carbocycles having 3 to 12 carbons, including bicyclic cycloalkyl and tricyclic structures. Approved cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and the like. The terms "aryls" and "heteroaryls" refer to monocyclic and polycyclic unsaturated or aromatic ring structures, with "aryl" referring to those which are carbocycles and "heteroaryl" which refer to those which are heterocycles. Examples of aromatic ring structures include phenyl, naphthyl, 1,2,3,4-tetrahydronaphthyl, furyl, thienyl, pyrrolyl, pyridyl, pyridinyl, pyrazolyl, imidazolyl, pyrazinyl, pyridazinyl, 1,2,3-triazinyl, , 2,4-oxadiazolyl, 1, 3, 4-oxadiazolyl, lH-tetrazol-5-yl, indolyl, quinolinyl, benzofuranyl, benzothiophenyl (tylphthanyl), and the like. The radicals can be optionally substituted by one more appropriate substituents, for example, a substituent selected from a halogen (F, Cl, Br and I); lower alkyl; OH; DO NOT?; CN; C02H; O-lower alkyl; aril; aryl-lower alkyl; CO- > CH ,; CONH-; OCH? CONH ?; NH ,; SO-NH ,; OCHF ?; CF ,; OCF3; and similar. The radicals can also be optionally substituted by a fused or bridged ring structure, for example OCH2-0. The term "aryl-lower alkyl" means a lower alkyl that produces an aryl. Examples include benzyl, phenethyl, pyridyl ethyl, naphylmethyl, and the like. The aryl-lower alkyl. it can be optionally substituted. In general, the various radicals or functional groups for X], X- > , Y and Z in Formula I may optionally be substituted by one or more appropriate substituents. Exemplary substituents include a halogen (F, Cl, Br, or I), lower alkyl, -OH, -N02, -CN, -CO-H, -O-lower alkyl, -aryl, -aryl-lower alkyl, - CO ^ CH ^, -C0NH2, -OCH2CONH2, -NH2, -S02NH2, haloalkyl (e.g., -CF3, -CH2CF3), -O-haloalkyl (e.g., -OCF3, -OCHF2), and the like. "Protection groups" refer to groups that protect one or more Inherent functional groups from the premature reaction. Appropriate protection groups can be routinely selected by those skilled in the art in clarity of the particular functionality and chemistry used to construct the compounds. Examples of appropriate protection groups are described, for example, in Greene and Wutz, Proteczing Groups in Organi c Synthesi s, 2nd edition, John Wiley i * Sons, New York, New York (1991). Exemplary protecting groups useful in the practice of the invention include tert-butoxycarbonyl (Boc) and the like. The compounds of the invention useful as GnRH agents are of the following Formula I: (I) wherein Z is a group selected from alkyl, alkenyl, alkynyl, cycloalkyl, heterocycle, aryl, heteroaryl, CH20R, and unsubstituted and substituted C (0) OR, where R is alkyl, alkenyl, alkynyl, cycloalkyl, aryl, or substituted or unsubstituted heteroaryl, and wherein the total number of carbon atoms present in Z, does not include optional substituents, in the range of 1 to 12; Y is a lipophilic group selected from alkyl, alkenyl, alkynyl, cycloalkyl, aryl, and unsubstituted and substituted heteroaryl, where the total number of carbon atoms present in Y, does not include optional substituents, ranging from 6 to 20.; Xi is a structural, or more spatial, unit used to connect the functional units CH_NC (0), Xi, Y, and Z in the three-dimensional space, which is selected from alkyl, alkenyl, alkynyl, cycloalkyl, aryl, and substituted heteroaryl and unsubstituted such that the atom is in the chain portion of the unit that binds to the central nitrogen (the N atom in Formula I is referred to as the "central nitrogen" to avoid confusion with any other nitrogen-produced substituent) and X2 ranges from 3 to 8, preferably from 4 to • 6; and X2 is a basic group having a pKa greater than about 8, preferably selected from a guanidinyl, amidinyl, acyloamidinyl, acetidinyl, and unsubstituted and substituted amino group; and pharmaceutically acceptable salts, prodrugs, pharmaceutically active metabolites, and mimomeric forms of the compounds. Additionally, Formula I is thought to cover, where applicable, the solvated as well as insolvated forms of the compounds. Thus, Formula I includes compounds having the indicated structure, including hydrated as well as non-hydrated forms. Preferred Z groups include: substituted and unsubstituted furyl, for example, 2-furyl, 3-furyl, 5-methyl-2-furyl, and 5-nitro-2-furyl; unsubstituted substituted pyrrolyl; unsubstituted substituted naphthyl, for example, 1-naphthyl and 2-naphthyl; substituted and unsubstituted thienyls, for example, 2-thienyl; substituted and unsubstituted pyridyls, for example, 2-chloropyrid-5-yl and 2-methylthiopyrid-3-yl; substituted and unsubstituted cycloalkyls, for example, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl; and substituted and unsubstituted lower alkyls, such as methyl, ethyl, propyl, butyl, and the like, with that substituted fluorine, for example, with CF3 being especially preferred. Preferred Z groups especially include furyl, pyrrolyl, thienyl, and CF5. Preferred Y groups include: unsubstituted substituted phenyl, for example 4-isopropyl phenyl, 4-N, N-dimethylaminopropyloxyphenyl, 2, 5-triethoxyphenyl, 2,3-dibenzyloxyphenyl, and 2- (4'-chlorophenyloxy) phenyl; unsubstituted substituted naphthyl and its "partially or fully saturated derivatives, eg, 1,2,3,4-tetrahydro-1,4,4,6-pentamethylnaphthalene; unsubstituted substituted dibenzylfuryl and its saturated derivatives; saturated, substituted and unsubstituted quinolinyl, isoquinolinyl, quinoxalinyl, and the like, for example, 3-quinolinyl, and substituted and unsubstituted cycloalkyls and fused polycyclic alkyls. Lipophilic groups which are bulky are particularly preferred of the Y groups, especially one of The following groups: Preferred groups Xi include alkylene, alkylene-cycloalkyl-alkylene, and alkylene-aryl-alkylene chains, where the term "alkylene" refers to (CH2) n, where n is an integer ranging from 1 to 6. Especially Preferred groups Xi include the following: (CH2) 4- - (CH2) 5-, Preferred X2 groups are very basic - for example, they have a pKa greater than about 9, more preferably greater than about 10. The X groups? examples include guanidinyl, amidinyl, and amino, which can be unsubstituted or substituted with one or more lower alkyls. Especially preferred for X2 is guanidinyl. Preferred compounds include those comprised by the following sub-generic Formulas I 'and I "(where Y and Z are as defined above): Preferred specific embodiments of compounds of Formula I include Compounds Nos. 1-1Z0 (in the following description, the alkyl and alkylene substituents are shown using a convention of abbreviations, for example, "for a methyl group): 6 7 22 24 34 35 36 37 38 39 101 103 104 105 106 107 108 1 18 119 120 Of the specific compounds identified above, Compounds Nos. 4, 5, 13, 25, 26, 30, 40, 47, 48, 55, and 80 are especially preferred. Other especially preferred GnRH agents of Formula I include the following Compounds Nos. 121-124: 121 122 123 124 As indicated above, the GnRH agents according to the invention also include active tautomeric and stereoisomeric forms of the compounds of Formula I, which can be obtained easily using techniques known in the art. For example, the optically active (R) and (S) isomers can be prepared by means of a stereospecific synthesis, for example, using chiral and chiral synthetic reagents, or racemic mixtures that can be resolved using conventional techniques. The GnRH agents further include multivalent or omitic forms of active forms of the compounds of Formula I. "Multimers" can be made by joining or placing multiple copies of an active compound in close proximity to each other, for example, using a scaffold provided by a carrier radical. Multimers of various dimensions (ie carrying varying numbers of copies of an active compound) can be tested to arrive at a multimer of optimal size with respect to the receptor link. The provision of multivalent forms of the active receptor binding compounds with optimal space between the receptor binding moieties can strengthen the receptor link (see, for example, Lee et al., Biochem., 1984, 23: 4255).

The skilled person can control the multivalency and spacing by selection of an appropriate carrier radical or linking units. Useful radicals include molecular supports that contain a multiplicity of functional groups that can react with functional groups associated with the active compounds of the invention. A variety of carrier radicals can be used to make highly active multimers, including proteins such as BSA (bovine serum albumin) or HAS, peptides such as pentapeptides, decapeptides, pentadecapeptides, and the like, as well as non-biological compounds selected for their beneficial effects, in absorbability, tra.usporte, and persistence within the target organism. The functional groups in the carrier radical, such as a.ro.inlo, sulfh.id.rilo, hydroxy "lo, -ya] quila-ni groups, can be selected to obtain stable bonds for the compounds of the invention, optimal space between the immobilization compounds, and optimal biological properties.Additionally, the GnRH agents of the invention include pharmaceutically acceptable salts of the compounds of the Formula I. The term "pharmaceutically acceptable" refers to the salts forms which are pharmacologically acceptable and substantially non-toxic to the subject being administered the GnRH agent .. Pharmaceutically acceptable salts include conventional acid addition salts or base addition salts formed of non-toxic organic or inorganic acids or appropriate inorganic bases. Exemplary acid addition salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, iodhydride acid ico, sulfuric acid, sulfamic acid, phosphoric acid, and nitric acid, and those derived from organic acids COJDO p-toluensujón fónj acid, methanesulfónic acid, ethane-disulfonic acid, isethionic acid, oxalic acid, p-bromophenylosulfonic acid, acid carbonic, succinic acid, citric acid, benzoic acid, 2-acetoxybenzoic acid, acetic acid, phenylacetic acid, propionic acid, glycolic acid, stearic acid, lactic acid, malic acid, tartaric acid, ascorbic acid, maleic acid, hydroxymalonic acid, acid glutamic acid, salicylic acid, sulphanilic acid, and fumaric acid. Exemplary base addition salts include those derived from ammonium hydroxides (for example, a quaternary ammonium hydroxide such as tetramethylammonium hydroxide), those derived from inorganic bases such as alkaline earth metal or alkaline hydroxides (eg, sodium, potassium, lithium, calcium or magnesium), and those derived from organic bases such as amines, benzylamines, piperidines, and pyrrolidines. The term "prodrug" refers to a metabolic precursor of a compound of Formula I (or a salt thereof) that is pharmaceutically acceptable. A prodrug may be inactive when administered to a subject but converted in vivo into an active compound of Formula I. The term "active metabolite" refers to a metabolic product of a compound of Formula I that is pharmaceutically acceptable. and cash. The prodrugs and active metabolites of the compounds of Formula I can be determined using techniques known in the art. A variety of assays and known techniques can be employed to determine the level of activity of various forms of the compounds in the GnRH system. Ligand-binding assays are used to determine the interaction with the receptor of interest. Where the link is of interest, a labeled receptor may be used, where the label is a fluorescer, an enzyme, a radioisotope, or the like, which records a quantifiable change in the receptor binding. Alternatively, the skilled person can provide an antibody to the receptor, where the antibody is labeled, which can be allowed for signal amplification. The link can also be determined by competitive displacement of a ligand bond to the receptor, where the ligand is labeled. with a detectable label. Where the agonist and / or antagonist activity is of interest, an intact organism or cell can be studied, and the change in an organismic or cellular function can be measured in response to the binding of the compound of interest. Various devices are available to detect the cellular response or a microphysiometer available from Molecular-Devices, Redwood City, California. In vi tro and in vi vi assays useful in measuring GnRH antagonist activity are known in the art. See, for example, Bowers et al., "LH supression in cultured rat pituitary cells trated whith 1 ng of LHRH," Endocrinol ogy, 1980, 1 6: 675-683 (in vi t ro,) and Corbin et al., "Antiovulatory active ty (AOA) in rats, "Endocr. Res. Commun. 1975, 2: 1-23 (in vi vo). The particular test protocols that can be used are described below. For example, GnRH receptor antagonists can be evaluated functionally by measuring the change in extracellular acidification rates as indicated below. The ability of the compounds to block the extracellular rate of acidification mediated by GnRH in HEK 293 cells of expression of human GnRH receptors is determined as a measure of the antagonist activity of the compound in vi tro. Approximately 100, 000 cells / chamber are immobilized in an agarose suspension medium (Molecular Devices) and perfused with a MEM medium without buffer using the Cytosensor® Microfiometer (Molecular Devices). The cells are allowed to equilibrate to the rate of basal acidification remaining stable (approximately one hour). The control dose response curves are performed at GnRü (10_11M to 10 ~ 7M). The compounds are allowed to incubate for 15 minutes prior to stimulation with GnRH, and are emitted for the antagonist activity. After incubation with the test compounds, the repetition of the dose response curves to GnRH is obtained. in the presence or absence of various concentrations of the test compounds. The Schild regression analysis was performed on the compounds to determine that if the compounds are opposed to GnRH-mediated increases in extracellular acidification rates through a competitive interaction with the GnRH receptorSee, for example, the acidification response using Antide peptide antagonist (CAS No. 11258-12-4) shown in Figure 2 (Antide was purchased from Sigma, Product No. A8802 (acetyl-B- (2 -naphthyl) -D-Ala-Dp-chloro-Phe-B- (3-pyridyl) -D-Ala-Ser-Ne- (Nicotinoyl) -Lys-Ne- (Nicotinoyl) -D-Lys-Leu-Ne- (I.soprnp-il) -Ty -Pro-D-? L a-NH ..)). The acidification response of the cells to GnRH is shown in the curves in Figure 2, where curve A is for the first exposure, curve B is for the second exposure, and curve C is for the third exposure. The GnRH responsive cell in the presence of 30nM Antide is shown in curve D of Figure 2. In another test, the accumulation of total inositol phosphates can be measured by the extraction of formic acid from the cells, followed by separation of the phosphates in the Dowex columns. The cells are divided using trypsin in two dishes of i 9 n or -Tiis G G? H? C: H-ioinositol (0.5 c-2 roCi per ml) for 16-18 hours in an inositol-free medium. The medium is then aspirated and the cells rinsed with either IX HBSS, 20 mm HEPES (pH 7.5), or serum free DMEM, IX HBSS, 20 mM HEPES (pH 7.5) containing the agonist, and 20 mM LiCl is then added and the cells are incubated for the desired time. The medium is aspirated and the reaction is stopped by the addition of lOmM of ice-cold formic acid, which also serves to extract cellular lipids. The inositol phosphates were separated by ion exchange chromatography on the Dowex columns, which are washed then with 5 ml of lOmM of myoinositol and lOmM of formic acid. The columns are then washed with 10 ml of 60 mM sodium format and 5 mM borax, and the total inositol phosphates are eluted with 4.5 ml of 1M ammonium formate, O. formic acid. Preferred GnR.H agents of the invention include those having a value of £ 7 of about 10 μM or less.The especially preferred GnRH agents are those which have a KA value in the nanomolar range.

Pharmaceutical Compositions The pharmaceutical compositions according to the invention comprise a suppressive amount of GnRH effective of at least one GnRH agent according to the invention and a pharmaceutically acceptable carrier or diluent or i e. These compositions can be prepared in a single dosage form appropriate for the desired mode of administration, for example, ? * > parenteral n oral To treat diseases or conditions mediated by GnPTH agonism or antagonism, a pharmaceutical composition of the invention is administered in an appropriate prepared formulation by combining a therapeutically effective amount (i.e., an effective GnRH modulating amount to achieve therapeutic efficiency). ) of at least one GnRH agent of the invention (as an active ingredient) with one or more pharmaceutically appropriate carriers or diluents. The formulations may be prepared according to conventional procedures, for example, by mixing, granulating, and properly compressing or dissolving the ingredients in known manners. Optionally, one or more different active ingredients, such as different GnRH antagonists, can be employed in a pharmaceutical composition. The pharmaceutical carrier can be a solid or liquid. Exemplary solid carriers include lactose, sucrose, talc, gelatin, agar, pectin, acacia. magnesium stearate, stearic acid, and the like. Illustrative of liquid carriers are syrup, peanut oil, olive oil, water, and the like. Simultaneously, the carrier or diluent may include materials of time delay or release time known in the art, such as glyceryl monostearate or glyceryl distearate, alone or in combination with a wax, ethylcellulose, hydroxypropylmethylcelluloses, methylmethacrylate, or Similar . A variety of pharmaceutical forms can be used. For example, if a solid carrier is used, the preparation may be in the form of a tablet, hard gelatin capsule, powder, pellet, troche, or pellets. The amount of solid carrier can vary widely, with an exemplary amount ranging from about 25 mg to about 1 g. If a liquid carrier is used, the preparation can be in. the form of a syrup, emulsion, soft gelatin capsule, sterile injectable solution, suspension in an ampoule or vial, or non-aqueous liquid suspension. "- To obtain a stable, water soluble dosage form, a pharmaceutically acceptable salt of a compound of Formula I can be dissolved in an aqueous solution of an organic or inorganic acid, such as 0.3M of a succinic acid solution or, more preferably, citric acid If "a soluble salt form is not available, the agent can be dissolved in one or more appropriate cosolvents. Examples of suitable cosolvents include alcohol, propylene glycol, polyethylene glycol 300, polysorbate 80, glycerin, and the like in concentrations ranging from 0% to 60% of the total volume. In an exemplary embodiment, a compound of Formula I is dissolved in DMSO and diluted with water. The composition may also be in the form of a solution of a salt form of a compound of Formula I in an appropriate aqueous vehicle, such as water, or isotonic or dextrose saline solutions. The pharmaceutical compositions of the present invention can be made using conventional techniques, for example, mixing, dissolving, granulating, dragee-making, pulverizing, emulsifying, encapsulating, entrapping, or lyophilizing processes. Pharmaceutical compositions can be formulated in a conventional manner using one or more physiologically acceptable carriers comprising excipients or auxiliaries selected to facilitate the process of the active compounds in the pharmaceutical preparations. An appropriate formulation is selected in view of the selected administration route. To prepare injectable preparations, the agents of the invention can be formulated in aqueous solutions, preferably in physiologically compatible buffers such as Han's solution, Ringer's solution, or saline-physiological buffer. For transmucosal administration, the appropriate penetrants for the barrier to be penetrated are used in the formulation and can be selected from those known in the art. For oral administration, the agents can be formulated rapidly by combining the active ingredient (s) with pharmaceutically acceptable carriers known in the art. The carriers allow the compounds of the invention to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, watered pastes, suspensions, and the like, for oral ingestion through a patient to be treated. Pharmaceutical preparations for oral use can be obtained by combining one or more agents with a solid excipient, optionally grinding the resulting mixture into granules, and processing the granule mixture after adding the appropriate auxiliaries, if desired, to get centers of tablets or dragees. Suitable excipients include fillers such as sugars (e.g., lactose, sucrose, mannitol, or sorbitol) and cellulose preparations (e.g., corn starch, wheat starch, rice starch, potato starch, gelatin, gum, cellulose methyl, hydroxypropylmethylcellulose, sodium carboxymethylcellulose, and / or polyvinylpyrrolidone (PVP)). If desired, disintegrating agents may be added, such as cross-linked PVP, agar, or alginic acid or a salt thereof such as sodium alginate. Dragee centers are provided with the appropriate coatings. For this purpose, concentrated sugar solutions can be used, which may optionally contain gum arabic, PVP, Carbopol ™ gel, polyethylene glycol, titanium dioxide, lacquer solutions, and / or one or appropriate organic solvents. Dyestuffs or pigments may be added for the coatings of the tablets or dragees for identification or to characterize the different combinations of the doses of the active compound. Pharmaceutical forms that are suitable for oral administration include soft-fit capsules made of gelatin, as well as sealed, soft capsules made of gelatin and a plasticizer, such as glycerol or sorbitol. The soft-fit capsules may contain the active ingredient (s) in a mixture with one or more fillers such as lactose, binders such as starches, and / or lubricant such as talc or magnesium stearate, and, optionally, stabilizers. In soft capsules, the active compound can be dissolved or suspended in a suitable liquid, such as fatty oil, liquid paraffin, or liquid polyethylene glycol. In addition, stabilizers can be added. For buccal administration, the compositions may take the form of tablets or lozenges formulated in a conventional manner. For administration by inhalation, the compounds to be used according to the present invention can be appropriately released in the form of an aerosol spray presentation of pressurized packings or an atomizer, with the use of an appropriate impeller, for example, dichlorodifluoromethane, trichlorofluoromethane , dichlorotetrafluoroethane, carbon dioxide, or other appropriate gas. In the case of a pressurized aerosol, the dosage unit can be determined to provide a valve to release a measured quantity. Capsules and cartridges, for example, gelatin for use in an inhaler or. insufflator can be formulated containing a powder mixture of the agent and a suitable powder base such as lactose or starch. The agents can be formulated for parenteral administration by injection, for example, by bolus injection or continuous infusion. Formulations for injection may be prepared in unit dosage form, for example, in ampoules, or in multi-dose containers, with an additional condom. The compositions may take the forms as suspensions, solutions, or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing, and / or dispersing agents. Pharmaceutical formulations for parenteral administration include aqueous solutions of the active compounds in water soluble form. Additionally, suspensions of the active compounds may be prepared as appropriate oily injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, or synthetic fatty acid esters such as ethyl oleate or triglycerides, or liposomes. Aqueous injectable suspensions may contain substances that increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol, or dextran. Optionally, the suspension may also contain stabilizers of appropriate agents that increase the solubility of the compounds to allow the preparation of highly concentrated solutions. Alternatively, the active ingredient may be in powder form for constitution with an appropriate vehicle, eg, sterile pyrogen-free water, before use. The compounds can also be formulated as rectal compositions, such as suppositories or retention enemas, for example, containing conventional suppository bases such as cocoa butter or other glycerides. In addition to the formulations described above, the compounds can also be formulated as a warehouse preparation. Long-acting formulations can be administered by implantation (e.g., subcutaneous or intramuscular) or by intramuscular injection. For example, the compounds can be formulated with suitable polymeric or hydrophobic materials (for example, as an emulsion in an acceptable oil) or ion exchange resins, or as sparingly soluble derivatives, for example, as a sparingly soluble salt. A pharmaceutically exemplary carrier for the hydrophobic compounds of the invention is a cosolvent system comprising benzyl alcohol, a non-polar surfactant, a water-miscible organic polymer, and an aqueous phase. The co-solvent system can be from the VPD co-solvent system (VPD is a solution of 3% by weight of benzyl alcohol, 8% by weight of the non-polar surfactant of polysorbate 80, and 65% by weight of polyethylene glycol 300, prepared for the volume in absolute ethanol). The VPD co-solvent system (VPD: 5W) is comprised of VPD diluted 1: 1 with 5% of a water solution in dextrose. This cosolvent system dissolves the hydrophobic compounds well, and the resulting formulation produces low toxicity in the systemic administration. As will be clear, the proportions of an appropriate co-solvent system can be varied in the clarity of the solubility and toxicity characteristics. In addition, the identity of the co-solvent components can be varied: for example, other non-polar low-toxic surfactants can be used instead of the polysorbate 80; the size of the polyethylene glycol fragment can be varied; one or more of the biocompatible polymers (eg, PVP) can be added or replaced by polyethylene glycol; and other sugars or polysaccharides can be replaced by dextrose. Alternatively, other delivery systems for the hydrophobic pharmaceutical compounds may be employed. Liposomes and emulsions are known examples of vehicles or release carriers for hydrophobic drugs and can be used to formulate the appropriate preparations. Certain organic solvents such as dimethyl sulfoxide can also be used, although this can cause an increase in toxicity. Additionally, the release can be achieved using a sustained release system, such as semipermeable matrices of solid hydrophobic polymers containing the therapeutic agent. The various sustained release materials are available and known to those skilled in the art. Sustained-release capsules can, and depend on their chemical nature, release the compounds for a lasting period of a few weeks or more than 100 days. Depending on the chemical nature and the biological stability of the therapeutic agent, additional techniques can be easily employed for. the stabilization of the protein. The pharmaceutical compositions may also comprise suitable solid or gel phase carriers or excipients. Examples of the carriers or excipients include calcium carbonate, calcium phosphate, sugars, starches, cellulose derivative, gelatin, and polymers such as polyethylene glycols. Some of the compounds of the invention can be provided as salts with pharmaceutically sharable ionic counters. Pharmaceutically acceptable salts can be formed with various acids, including hydrochloric, sulfuric, acetic, lactic, tartaric, malic, succinic acids, and the like. The salts tend to be more soluble in aqueous or other protic solvents, which are the corresponding free base forms. It will be appreciated that the actual dosages of agents used in the compositions of this invention will vary according to the particular complex to be used, the particular composition formulated, the mode of administration, and the particular site, host, and disease being treated. Optimal dosages for a given group of conditions can be determined by those skilled in the art using conventional tests of dosage determination in view of the experimental data for a given compound. For oral administration, an exemplary daily dose generally employed will be from about 0.001 to about 1000 mg / kg per body weight, with courses of treatment repeated at appropriate intervals. The administration of the prodrugs can be dosed at weight levels that are chemically equivalent to the weight levels of the fully active compounds. Examples of the specific pharmaceutical preparations according to the invention are given below.

Parenteral Composition: To prepare a pharmaceutical composition of this invention suitable for administration by injection, 100 mg of a pharmaceutically acceptable water soluble salt of a compound of Formula I are dissolved in DMSO and then mixed with 10 ml of 0.9% sterile saline solution. The resulting mixture is incorporated into a unit dosage form suitable for administration by injection. Oral composition: To prepare an orally administrable pharmaceutical composition, 100 mg of a compound of Formula I was mixed with 750 mg of lactose. The resulting mixture was incorporated in a single dosage form suitable for oral administration, such as a hard gelatin capsule.

Synthesis of Intermediate GnRH Agents The compounds of Formula I that can be advantageously processed use intermediates according to the invention. In particular, the intermediates of the invention are of the following Formulas II, III, and IV: where: j is 1 or 2; k is 1, 2, 3, 4, or 5; Rs is H or substituted or unsubstituted lower alkyl; R9 is H or lower alkyl, CN, NO, or CO2R1 substituted or unsubstituted; R10 is H or lower alkyl, CH-2OR1; (CHzJpOR1; CO2R1; or (CH2) C (0) R2 substituted or unsubstituted, where p is an integer from 1 to 6, and R2 is H, OR1, SR1, NIR1) -;, or C (R1) 3, R11 is H or lower alkyl, CH20-phenyl, CH20-benzyl, phenyl or benzyl substituted or unsubstituted; or any two of R8, R9, R10, and R11 together form a 5- or 6-membered heterocycle; and each R1 is independently selected from H and substituted or unsubstituted lower alkyl, O-lower alkyl, and S-lower alkyl, with tBu which is in the preferred R1 group. Preferred intermediaries include: Intermediate compounds have utility in forming the X? ~ X2 portion of the compounds of the present invention, by reacting the amine group with an aldehyde to form a secondary amine, which can then be treated with acyl to form the compounds of the Formula I general.

Scheme of reaction. The compounds of Formula 1 can be prepared according to the following general reaction scheme, wherein the spherical symbol represents the radical (CH) j, of cyclopentyl, cycloexyl, or benzyl of Formula II, III, or IV: To illustrate, the synthesis described in the reaction scheme below employs the intermediates of the Formula. II to prepare the compounds of Formula I wherein the unit of X1-X2 is 1-guanidinomethyl-4-aminomethylcyclohexyl. As described in more detail below, stage 1 involves the protection of the basic group, using ter-methoxycarbonyl (Boc) as a protection group. In the illustrated scheme, the basic group is guanidinyl, but, as it will be readily apparent to experts, this reaction is also applicable to other basic groups such as amines and amidines. Step 2 involves the formation of a secondary amine by reaction with the ZCHO aldehyde, where Z is as defined above. Stage 3 involves acylation of the secondary amine with YCOC1, where Y is as defined above. Stage 4 involves the deprotection of the basic group by acid hydrolysis. The product can be isolated as the corresponding salt.

Step 1-Preparation of the Protected Compound by 1- (N, N'-diBoc) -guanidinomethylation: Alternative steps 1 (A) and 1 (B) below provide two general procedures of 1- (N, N '-diBoc) -guanidinomethylation. Step 1 (A): For a solution of diamine (2.00 mmol equivalent) in THF (0.7M) is added a solution of 1-H-pyrazole-1 - (N, -bis (terbutoxycarbonyl) carboxamidine) (1.00 mmol equivalent) in THF (0.7M). The solution is stirred at room temperature for 3 hours (h), or until no further transformation can be observed by TLC (thin layer chromatography). The solvent is removed under reduced pressure to provide a syrupy residue, which is started in ethyl acetate (-1.5 times the amount of the volume of THF used in the reaction or the volume of solvent needed to dissolve the amount of residue obtained) and washed with water to neutral pH. The organic layer is washed with brine, dried over MgSO4, and concentrated. The product is purified by column chromatography on silica gel and eluted or eluted with an appropriate elution solvent (which can be determined rapidly, for example, using 5% MeOH in dichloromethane as a starting point). The solvents are removed in vacuo to allow 1- (N, N '-diBoc) -guanidino ethyl-linked-amine. In addition, other reagents can be used to place a N, N'-diBoc-guanidine unit protected from diamines, such as 1,3-bis (tert-butoxycarbonyl) -2-methyl-2, thiopseudourea (CAS No. 107819-90-0). ). Alternatively, 1-H-pyrazole-1- (N, N-bis (terbutoxycarbonyl) carboxamidine) can be added directly as a solid, rather than as a solution as described above. Step 1 (B): For a solution of diamine (1.00 mmol equivalent) in THF (0.07M) is added its own portion as a solid (for a period of time of 10 minutes) 1-H-pyrazole-1- (N, N-bis (tert-butoxycarbonyl) carboxamidine) (1.00 mmol equivalent). The solution is stirred at room temperature for 0.5 hour. The solvent is removed under reduced pressure to provide a syrupy residue that is initiated in ethyl acetate (0.5 times the volume amount of THF used in the reaction, or the volume of solvent needed to dissolve the amount of residue obtained) and washed two times with water.

The layers are separated, and the product is purified by column chromatography on silica gel and levigated with 100% ethyl acetate to remove any non-polar impurities and then with 100% isopropyl alcohol to provide the pure product. The solvents are removed in vacuo to leave the desired product. Normal TLC conditions are 15: 85: 0.1 methanol / chloroform / acetic acid. The normal range of yield ranges from 40% to 44% of the desired protected compound.

Stage 2- Reductive Mining: Reductive amination can be achieved in an appropriate manner. For the reductive amination of aldehydes and ketones with sodium triacetoxyborohydride, see generally: Abdel-Magid et al., J. Org. Chem; 1996, 61: 3849. Two alternative procedures of reductive inactions are described below.

Step 2 (A): 3, 5, 5, 8, 8-Pentamethyl-5, 6, 7, 8-tetrahydro-2-naphth-aldehyde (1.00 mmol equivalent) and 1- (N, N '-diBoc) - guanidinomethyl-union-amine (1.00 mmol equivalent) is dissolved in methanol (0.09M). Then, 1% glacial acetic acid is added to the methanol solution. (10% of the volume of methanol used) followed by NaCNBH3 (1.00 mmol equivalent), and the reaction contents are stirred overnight. The reaction is assayed by TLC to reveal three components (aldehyde, desired product, and starting guanidine derivative). The reaction is terminated by adding water (50% of the volume of methanol used), extracted with dichloromethane (10 times the volume of methanol used), and washing with saturated sodium bicarbonate. The organic layer is dried over magnesium sulfate, filtered, and concentrated. The product is purified by column chromatography on silica gel and levigadp with an appropriate elution solvent (eg, 3: 1 ethyl acetate in hexanes to remove the aldehyde without reaction, followed by elution with 1: 1 acetate. of ethyl in hexanes), obtaining the desired reductive amination product. In some cases, heating to reflux for 2 hours will facilitate the imine formation reaction. Also see, A. Abdel-Magid et al., J. Org. Chem. , 1996, 61: 3849, which describes the reductive amination of aldehydes and ketones with sodium triacetoxyborohydride. ~~ Stage 2 (3): 3, 5, 5, 8, 8-Pentamethyl-5, 6, 7, 8-tetrahydro-2-naphth-aldehyde (1.00 mmol equivalent) and 1- (N, N '-diBoc) ) -guanidinomethyl-union-amine (1.00 mmol equivalent) is dissolved in methanol (0.09M). Then, NaBH4 (1.00 mmol equivalent) is added (in ethanol by means of the additional small-scale procedures provided below, or carefully as a solid) and the reaction contents are stirred overnight. The reaction is assayed by TLC to reveal three components (aldehyde, desired product, and starting guanidine derivative). The reaction is terminated by adding water (50% of the volume of methanol used), extracted with dichloromethane (10 vec &s the volume of methanol used), and washing with saturated sodium bicarbonate. The organic layer is dried over magnesium sulfate, filtered, and concentrated. The product is purified by column chromatography on silica gel and eluted with an appropriate elution solvent or levigation (as can be easily determined by the skilled person, for example, with 3: 1 ethyl acetate in hexanes to remove the aldehyde without reaction followed by elution with 1: 1 ethyl acetate in hexanes), to obtain the desired reductive amination product. In some cases, heating under reflux for 2 hours will facilitate the reaction of imine formation.

Stage 3 - Acylation: The products of reductive amination (1.00 mmol equivalent) are dissolved in dichloromethane (-0.2 to 0 05M, depending on the solubilities of the substrates), followed by the addition of triethylamine (2.00 mmol equivalent) and sodium chloride. 2-furoyl (1.00 mmol equivalent). The reaction contents are stirred overnight at room temperature (RT). The mixed reaction is diluted with dichloromethane (5 times the amount of dichloromethane used) and washed with saturated sodium bicarbonate. The organic layer is dried over magnesium sulfate and filtered. The product is purified by column chromatography on silica gel and eluted with an appropriate elution solvent (e.g., 3: 1 ethyl acetate rhexanes). The solvents are removed in vacuo for the yield of the acylated product.

Step 4 - Deprotection of the Basic Group: The product of the acylation step (1.00 mmol equivalent) is dissolved in a solution of 25-50% TFA in dichloromethane (0.02M), and the reaction contents are stirred at room temperature (15-20 minutes, the solution becomes slightly orange-red). The reaction contents are stirred for an additional 1 hour- and 20 minutes or until BOC deprotection is complete. The reaction is terminated by concentration in vacuo, followed by the addition of water / acetonitrile (0.006M) and lyophilization overnight. The final compound is purified by liquid chromatography methodology. high performance (HPLC). The solvents are removed in va cuo (yields ranging from 30% to 50%) to give the product. An alternative procedure for the removal of the N, N'-bis-BOC guanidines using tin tetrachloride, which can provide the corresponding guanidinium chloride salts, is described in Miel et al., Tetrahedron Letters, 1997, 38: 7865- 7866 Preparation of Reagents: Reagents useful for synthesizing the compounds can be obtained or prepared according to techniques known in the art. For example, amine-free preparation from the common salt forms and stored reagent solutions may be useful for small scale reactions. Also see Abdel-Magid • et al., "Reductive ination of aldehydes and ketones with sodium triacetoxyborohydride," J. Org. Chem. , 1996, 61: 3849. The methanolic solutions of the free bases may be prepared from chlorhydride, chlorhydride, hydroperoxide, or other salts when the free base is soluble in methanol. In this procedure, once sodium methoxide is added, care must be taken to prevent exposure to air, since amine-free bases, particularly primary amines, absorb carbon dioxide from the air to form salts. An amount of 10-ml of a 0.1 M solution of a free base in methanol can be prepared as indicated below. The weight of 1.0 mmol of a monochlorhydride salt in an untapped Erlenmeyer flask containing a stirred bar, and add 7 ml of methanol. To the stirred suspension, add 229 ml (1.0 mmol equivalent) of sodium methoxide in methanol (25 weight percent, 4.37M), cap the flask, and stir the mixture vigorously for 2 hours. The suspension will sometimes change appearance as a precipitated, thinner sodium chloride milk forms. Filter the suspension through a 15-ml fragmented glass funnel medium, wash the filter case with 1-2 ml of methanol, transfer the filtrate to a 20-ml flask, and dilute to 10 ml with methanol. The theoretical yield of sodium chloride is approximately 59 mg, but the recovery is usually not quantitative, leading to a slight solubility in methanol. For a hydrochloride salt, a second equivalent of sodium methoxide (458 ml) is required. A 0.5M solution of sodium borohydride in etarrol can be prepared as follows. Sodium borohydride (520 mg, 13.8 mmol) is stirred pure anhydrous (non-denatured) ethanol (25 ml) for ~ 2-3_ minutes. The suspension is filtered through a fragmented glass funnel medium to remove a small amount of undissolved solid (typically about 5% or 25 mg of the total borohydride mass). The filtrate must appear as a discolored solution that only evolves a little hydrogen. This solution must be used immediately, since it decomposes significantly over a period of a few hours, producing the formation of a gelatinous precipitate. Sodium borohydride is hygroscopic, to avoid exposure to air the solution is made immediately after weighing the solid. Sodium borohydride has a solubility of about 4% in ethanol at room temperature. This corresponds to a little more than 0.8M. However, sometimes a small percentage of the solid remains undissolved without taking into account the concentration to be prepared, even after being stirred during >; 5 minutes. To perform the small scale synthesis of compounds of Formula I, the reactions described below can be performed to prepare several reactants useful in the reaction scheme described above. As with the rest of the specification, all temperatures in the following description are in degrees Celsius and all parts and percentages are by weight, unless otherwise indicated. The starting materials and other reagents used in the following examples of commercial suppliers, such as Áldrich Chemical Company or Lancaster Synthesis Ltd., may be purchased and used without further purification, unless otherwise indicated. Tetrahydrofuran (THF) and N, N-dimethylformamide (DMF) are obtained from Aldrich in SureSeals ® bottles and used as received. All solvents are purified using standard methods in the art, unless otherwise indicated. The reactions indicated below are carried out under a positive nitrogen pressure or with a drying tube, at room temperature (unless otherwise indicated), in anhydrous solvents, and the reaction flasks are fixed with a rubber septum. the introduction of substrates and reagents by means of a syringe. The glassware is kiln dried and / or heat dried. Analytical thin-layer chromatography (TLC) is performed on plates 254 to 60 ° F of glass silica gel with backing (Analtech (0.25 mm)) and levigated with the appropriate proportions of solvents (v / v). The reactions are assayed by TLC and terminated as judged by the consumption of starting material. The tip plates are visualized with a p-anisaldehyde spray reagent or phosphomolybdic acid reagent (Aldrich Chemical, 20 weight percent ethanol) and heat activated. The hard work is usually done by doubling the reaction volume with the reaction solvent or extraction solvent and then washing with the indicated aqueous solutions using 25% by volume of the extraction volume (unless otherwise indicated). The product solutions are dried over anhydrous NaSO 4 before filtration, and the evaporation of the solvents is under reduced pressure in a rotary evaporator and noting how the solvents are removed in vacuo. Column chromatography at the moment. { Still and collaborators, A. J. Org. Chem. , 1978, 43: 2923) is directed using Baker's immediate grade silica gel (47-61mm) and a ratio of raw material: silica gel from about 20: 1 to 50: 1, unless otherwise noted. indicate Hydrogenolysis is done at the indicated pressure or at ambient pressure. The 1 H-NMR spectra are recorded on a Bruker instrument operating at 300 MHz, and the 13 C-NMR spectra are recorded operating at 75 MHz. The NMR spectra are obtained as well as the CDC13 solutions. (reported in ppmj using chloroform as a standard reference (7.25 ppm and 77.00 ppm) or CD3OD (3.4 and 4.8 ppm and 49.3 ppm), or a standard internal tetramethylsilane (0.00 ppm) when appropriate Other NMR solvents are used as appropriate. When the peak multiplicities are reported, the following abbreviations are used: s = single, d = doublet, t = "triplet, m = manifold, br = broad, dd = doublet of doublets, dt = doublet of triplets. of coupling, when given, - are reported in Hertz.The infrared spectra are recorded on a Perkin-Elmer FT-IR Spectrometer as pure oils, as pellets of KBr, or as solutions of - CDCI3, and when reported are in numbers Wavelength (cm-1) The mass spectra are obtained using LSIMS or electroroated, all the fusing points are not correct.

Preparation of the Link Block. 1-H-pyrazole-l-carboxamidine: _ ___ 1-H-pyrazole-1-carboxamidine is prepared according to Bernatowicz et al., J. Org. Chem. , 1992, 57: 24U7-2502 (and references therein), and protected with di-tert-butyl dicarbonate to give 1-H-pyrazole-1- (N, N-bis (tert-butoxycarbonyl) carboxamidine) of-agreement to Drake and collaborators, Syn th. , 1994, 579-582.

Preparation of 1- (N, N '-diBoc) -guanidinomethyl-4- to inomethylcyclohexane: To a solution of 1,4-bis-aminomethyl-cyclohexane (20 g, 0.14 mol) in THF (200 ml) is added a solution of 1-H-pyrazole-1 - (N, N-bis (tert-butoxycarbonyl) carboxamidine) (22.0 g, 0.07 mol) in THF (100 ml). (Note that 1-H-pyrazole-1- (N, N-bis (tert-butoxycarbonyl) carboxamidine) does not need to be dissolved in THF, rather it can be added pure as a solid to the process.) The solution is stirred at room temperature during 3 hours. The solvent is removed under reduced pressure to provide a syrupy residue, which is started in ethyl acetate (500 ml) and washed with water until neutral pH. The organic layer is washed with brine, dried over MgSO4, and concentrated. The product is purified by column chromatography on silica gel and eluted with 5% MeOH in dichloromethane. The solvents are removed in vacuo to leave 11.6 g (43% yield) of 1- (N, N'-diBoc) -guanidinomethyl-4-aminomethyl cyclohexane (Compound (a)). 1H-NMR (CDC13) d 11.5 (broad s, 1H), 8.35 (broad s, 1H), 3.26 (dt, 2H), 2.52 (dd, 2H), 1.82-0.97 (m, 28H, with a single to 1.5 ). An alternative preparation of 1- (N, N'-diBoc) -guanidinomethyl-3-aminomethylcyclohexane is as indicated below. For a solution of cis / trans 1,4-bis-aminsmethyl-cyclohexane (9.0 g, 63.3 mmol) in THF (903 ml, 0.07M) a portion of its own is added as a solid (for a period of 10 minutes) 1-H-Pyrazole-1- (N, N-bis (tert-butoxycarbonyl) carboxamidine) (19.6 g, 63.3 mmol.) The solution is stirred at room temperature for 0.5 hour. it is removed under reduced pressure to provide a syrupy residue, which is started in ethyl acetate (500 ml) and washed twice with water.The layers are separated and the product is purified by column chromatography on silica gel and levigated with 100% ethyl acetate to remove any non-polar impurities, followed by elution with 100 ° or isopropyl alcohol, to provide the pure product.The solvents are imidates in vacuo to leave 10.2 g (42% yield) of 1 - (N, N'-diBoc) guanidinomethyl-4-a-inomethylcyclohexane, 1H-NMR (CDC13) d 11.5 (broad s, 1H), 8.35 (broad s, 1H), 3.26 (dt, 2H), 2.52 (dd, 2H), 1.82-0.97 (m, 28H, with a single to 1.5).

ReductiveAmination: _. _. . " 3, 5, 5, 8, 8-Pentamethyl-5,6,7,8-tetrahydro-2-naphth-aldehyde (0.2021 g, 0.88 mmol) and 1- (N, N '-diBoc) -guanidinomethyl-4- aminomethylcyclohexane (Compound (a), 0.337 g, 0.88 mmol) is dissolved in methanol (10 ml). Then, the solution of 1% glacial acetic acid-in methanol (100 μl) is added followed by NaCNBH3 (55.4 mg, 0.88 mmol, 1.0 equivalent), and the reaction contents are stirred overnight. The reaction is assayed by TLC to reveal three components (aldehyde, desired product, and starting guanidine derivative). The reaction is terminated by the addition of water (-5 ml), extracted with dichloromethane (-100 ml), and washed with saturated sodium bicarbonate. The organic layer is dried over magnesium sulfate, filtered, concentrated, and eluted by column chromatography with 3: 1 ethyl acetate in hexanes to remove the unreacted aldehyde, followed by elution with 1 ml. 1: ethyl acetate in hexanes, yielding the desired product (Compound (b), cyclohexyl, cis / trans mixture). The solvents are removed in va cuo (normal range of normal yields of 50 to 80%).

Preparation of the Acylated Derivative. Followed by TO. Guanidine Deprotection: (d) The product of reductive amination (320 mg, 0.54 mmol, 1.0 equivalent) is dissolved in dichloromethane (10-15 ml), followed by the addition of triethylamine (129 mg, 1.08 mmol, 2 equivalent), and 2-furoyl (0.53 mmol, 52 μl, 1.0 equivalent). The reaction contents are stirred overnight at room temperature. The reaction is diluted with dichloromethane (50 ml) and washed with saturated sodium bicarbonate. The organic layer is dried over magnesium sulfate, filtered, and purified by column chromatography and eluted using 3: 1 hexanes in ethyl acetate. The solvents are removed in vacuo (the normal reaction generally yields a range of 50 to 80%) to give Compound (c). The product of the -acrylation reaction (Compound (c), 220 mg, 0.318 mmol, 1.0 equivalent) is dissolved in a 50% solution of TFA in dichloromethane (20-25 ml), and the reaction contents are stirred at room temperature (15-20 minutes, the solution becomes slightly orange-red). The reaction contents are stirred for an additional 1 hour and 20 minutes until deprotection is complete. The reaction is terminated by concentration in vacuo, followed by the addition of water / acetonitrile (-50 ml) and lyophilization overnight. The final compound is purified by HPLC methods. The solvents are removed in vacuo (yield range 30 to 50%) to give Compound (d). The above analysis is related to the preparation of Compounds (a) - (d). The following analysis relates to the preparation of Exemplary Compounds (e) - (k). The compounds (e) - (k) can be used as described above - to produce the corresponding deprotected compounds (guanidinyl free), through hydrolysis under acidic conditions.

Preparation of 1- (N, N '-diBoc) -gua.nidinomet Ll-3-aminomethylcyclohexane: (e) For a solution of cis / trans-l, 3-bis-aminomethylcyclohexane (7.5 g, 52.8 mmoi) in THF (30 ml) is added a solution of 1,3-bis (tert-butoxycarbonyl) -2- methyl-2-thiopseudourea (7.65 g, 26.3 mmol) in THF (40 mL) within 0.5 hour. The solution is stirred at room temperature for 5 hours. The solvent is removed under reduced pressure, and the product is purified by column chromatography on silica gel using a mixture of methylene chloride / methanol as eluent, to leave 2.2 g (22% yield) of 1- (N, N '-diBoc) -guanidinomethyl-3-aminomethylcyclohexane (Compound (e)). NMR-: H (CDC1 d 11.53 (broad s, 1H), 8.40 (broad s, 1H), 3.28-3.30 (m, 2H), 2.54-2.61 (m, 2H), 1.81 (broad s, 2H), 1.27 -1.58 (m, 26H), 0.89 (m, 1H) 0.65 (m, 1H) Alternatively, Compound (e) can be prepared as follows For a solution of ci s / trans 1,3-bis-aminomethylcyclohexane (10.0 g, 70.3 mmol) in THF (1000 mL, 0.07M) an own portion is added as a solid (over a period of 10 minutes) 1-H-Pyrazole-1- (N, N-bis (tert-butoxycarbonyl) carboxamidine) (21.8 g, 70.3 mmol ). The solution is stirred at room temperature for 0.5 hour. The solvent is removed under reduced pressure to provide a syrupy residue which is started in ethyl acetate (500 ml) and washed twice with water. The layers are separated, and the product is purified by column chromatography on silica gel and levigated with 100% ethyl acetate to remove any non-polar impurities, followed by elution with 100% isopropyl alcohol to provide the pure product. . The solvents are removed in vacuo to leave 11.4 g (41% yield) of 1- (N, N'-diBoc) -guanidinomethyl-3-aminomethylcyclohexane. 1H NMR (CDClj) d 11.53 (broad s, 1H), 8.40 (broad s, 1H), 3.28-3.30 (m, 2H), 2.54-2.61 (m, 2H), 1.81 (broad s, 2H), 1.27 -1.58 (m, 26HJ, 0.89 (m, 1H), 0.65 (mLH).

Preparation of 1- (N, N '-diBoc) -guanidinomethyl-4-aminomethylbenzene: For a solution of p-xylylenediamine (6.44 g, 47.4 mmol) in THF (30 ml) is added a solution of 1,3-bis (tert-butoxycarbonyl) -2-methyl-2-thiopseudourea (6.63 g, 22.9 mmol) in THF (40 ml) within 0.5 hour . The solution is stirred at room temperature for 5 hours. The solvent is removed under reduced pressure, and the product is purified by column chromatography on silica gel using a mixture of methylene chloride / methanol as the eluent, to leave 8.0 g (92% yield) of 1- (N , N '-diBoc) -guanidinomethyl-4-aminomethyl benzene (Compound (f)). NMR-XH (CDC13) d 11.54 (broad s, 1H), 8.56 (broad s, 1H), 7.29 (s, 4H), 4.60 (d, 2H), 3.86 (s, 2H), 1.64 (broad s, 2H) ), 1.52 (s, 9H), 1.48 (s, 9H).

Preparation of 1- (N, N'-diBoc) -guanidinomethyl-3-aminomethylbenzene: _,.

To a solution of m-xylylenediamine (7.14 g, 52.5 mmol) in THF (30 ml) is added a solution of 1,3-bis (tert-butoxycarbonyl) -2-methyl-2-thiopseudourea (7.57 g, 26.1 mmol) in THF (40 ml) within 0.5 hour. The solution is stirred at room temperature for 5 hours. The solvent is removed under reduced pressure, and the product is purified by column chromatography on silica gel using a methylene chloride / methanol mixture as the eluent, to leave 7.9 g (80% yield) of 1- (N , N '-diBoc) -guanidinomet il-3-aminsmethylbenzene (Compound (g)). 1H-NMR (CDCY) d 11.54 (broad s, 1H) 7 8.58 (broad s, 1H), 7.19-7.34 (m, 4H), 4.62 (d, 2H), 3.86 (s, 2H), 1.83 (s broad , 2H), 1.52 - (s, 9H), 1.48 (s, 9H).

Preparation = of, _ i- (N, N'-diBoc) -guanidine-4-aminobutane: ". (h) To a solution of 1,4-diaminobutane (4.15 gt ~ 47.1 g, 47.9 mmol) in THF (30 ml) is added a solution of 1,3-bis (tert-butoxycarbonyl) -2-methyl-2- tiopseudourea (6.83 g, 23.6 mmol) in THF (40 ml) within 0.5 hour. The solution is stirred at room temperature for 5 hours. The solvent is removed under reduced pressure, and the product is purified by column chromatography on silica gel using a mixture of methylene chloride / methanol as the eluent, to leave 3.0 g (40% yield) of 1- (N , N '-diBoc) -guanidino-4-aminobutane (Compound (h)). Y-NMR (CDC1) d 11.49 (broad s, lp), 8.35 (broad s, 1H), 3.42-3.47 (m, 2H), 2.72-2.76 (t, 2H), 0.86-1.65 (m, 24H). "~" An alternative procedure for preparing Compound (h) is as indicated below. For a solution of 1,4-diaminobutane (6.0 g, 68.1 mmol) in THF (972 mL, 0.07M) a portion of it is added as a solid (over a period of 10 minutes) 1-H-pyrazole-1- ( N, N-bis (tert-butoxycarbonyl) carboxamidine) (21.5 g, 68.1 mmol). The solution is stirred at room temperature for 0.5 hour. The solvent is removed under reduced pressure to give a syrupy residue which is started in ethyl acetate (500 ml) and washed twice with water. The layers are separated- and the product is purified by column chromatography on silica gel and eluted with 100% ethyl acetate to remove any non-polar impurities and then with 100% isopropyl alcohol to provide the pure product. The solvents are removed in vacuo to leave 10.0 g (44% yield) of 1- (N, N '-diBoc) -guanidino-4-aminobutane. NMR-: H (CDC1 d 11.49 (broad s, 1H), 8.35 (broad s, 1H), 3.42-3.47 (m, 2H)? 2.72-2.76 (t, 2H), 0.86-1.65 (, 24H).

Preparation of 1-N, N-dimethylaminpmethyl-4-amino-ethylbenzene For a solution of 1-N, -dimethylaminomethyl-4-carbonitrilbenzene (4.8 g, 30 mmol) in THF, a solution of 1 M borane tetrahydrofuran complex (90 ml) is added. The mixture is heated at reflux temperature for 16 hours under nitrogen. After cooling to room temperature, a solution of 1M HCl in methanol (100 mL) is added. The reaction mixture is refluxed for 3 hours. The product, which is precipitated, is collected by filtration, washed with diethyl ether, and dried in vacuo to give 5.9 g (83% yield) of the product as the hydrochloride salt (Compound (i)): NMR-XH ( DMSO-d d 8.65 (s broad 3H), 7.55 (dd, 4H), 4.25 (s, 2H), 3.98 (s, 2H), 2.62 (s, 6H). repair of 1- (N, N '-diBoc) -guanidinpmethyl-2- ^ __ aminemethylbenzene or) To a solution of o-xylylenediamine (7.14 g, 52.5 mmol) in THF (30 ml) is added a solution of 1,3-bis (tert-butoxycarbonyl) -2-metii-2-thiopseudourea _ (7.57 g, 26.1 mmol ) in THF (40 ml) within 0.5 hour. The solution is stirred at room temperature for 5 hours. The solvent is removed under reduced pressure, and the product is purified by column chromatography on silica gel using a mixture of methylene chloride / methanol as the eluent, to leave the (Compound (j)) 1- (N, N 'diBoc) -guanidinomethyl-3-aminomethylbenzene. Alternatively, Compound (j) can be prepared in a manner analogous to the alternative preparation described above for the Compound (e) Preparation of 1- N, N * -diBoc) -guanidinomethyl-2-aminomethylcyclohexane:. .

() For a solution of cis / trans-1,2-bis-aminomethylcyclohexane (7.5 g, 52.8 mmol) in THF (30 mL) is added a solution of 1,3-bis (tert-butoxycarbonyl) -2-methyl-2- tiopseudourea (7.65 g, 26.3 mmol) in THF (40 ml) within 0.5 hour. The solution is stirred at room temperature for 5 hours. The solvent is removed under reduced pressure, and the product is purified by column chromatography on silica gel using a methylene chloride / methanol mixture as the eluent, to leave (Compound (k)) 1- (N, N '-diBoc) -guanidinomethyl-2-aminomethylcyclohexane. Alternatively, Compound (k) can be prepared in a manner analogous to the alternative preparation described above for Compound (e). Preparation of Benzylamines: The following general reaction procedure provides a method for the preparation of benzylamines for use with aldehydes containing protected guanidines as an alternative to diamines. The reductive aminations proceed under the same reaction conditions for the protected guanidine amines described above.

To a solution of 6-claromethyl-1,3,4-tetrahydro-1,4,4-tetramethylnaphthalene (10 g, 42.2 mmol, 1 00 mmol, equivalent) in DMF (84 mL) is added sodium azide ( 13.7 g, 211.2 mmol, 5.00 mmol equivalent) at room temperature and the reaction contents are stirred overnight. The reaction is checked by TLC (5% ethyl acetate in hexanes;) The reaction is terminated by purifying the reaction contents in water and extracting with ethyl acetate.The organic solvents are removed in vacuo and the crude product is filtered through. from a plug of silica gel (ratio-10: 1) and extracted with hexanes.The solvent is removed in vacuo and stored under vacuum to leave 10 grams of the desired product of 6-azidomethyl-1,2,3. 4-Tetrahydro-1,2,4-tetramethiinaf alene (97% yield) For a solution of 6-azidomethyl-1,2,4,4-tetrahydro-1,1,1-tetramethylnaphthalene (9.0 g) , 37 mmol, 1.00 ml equivalent) in water / THF (1: 5 ratio, 0.16M, 231 me) is added polymeric triphenylphosphine (14.8 g, 44.38 mmol, 1.20 mmol equivalent, -3 mmol / g resin) at room temperature, and the reaction contents are stirred for 18-24 hours.The reaction is checked by TLC (5% ethyl acetate in hexanes). It is woven by filtering through a filter paper and washing the resin with methanol, and the product is verified by NMR ^ H. The organic solvents are removed in vacuo, and the product is filtered through a plug of silica gel (ratio of 10: 1) and eluted with 25% isopropyl alcohol in chloroform to remove a minor impurity of higher performance. The organic solvents are removed in vacuo, yielding a thick oil that solidifies upon standing (7.5 g, 93% yield). The chemical reactions described above have a general applicability to the preparation of GnRH agents of the invention. Thus, other GnRH agents can be similarly prepared by an appropriate modification as will be readily appreciated by those skilled in the art, for example, through the protection of interference groups, adapted for use with other conventional reagents, and / or by routine modifications of the reaction conditions.

Biological verification The compounds are tested for activity according to the following protocol.

Stable transfection of GnRH receptors Human . The cDNA for the human GnRH receptor is cloned into the vector of the expression plasmid, cDNA 3 (In Vitrogen), and stably transfected into HEK 293 cells (a cell line developed by Dr.

Stuart Sealfon, Mount Sinai Medica School, New York, New York), The human GnRH receptor (hGnRH) is cloned into cDNA 3 (In Vitrogen), which provides high level expression and marked ~ -omycin resistance. Lipofectamine (GIBCO BRL) is used to transfect HEK 293 cells with plasmid DNA. This method involves mixing the plasmid and the reagent in a serum free medium, allowing to form the DNA liposome complexes, and coating the cells with this solution. HEK 293 cells are electrodeposited (germinated) in a 10 cm tissue culture dish at a density of 3 x 10 e cells / dish the day before transfection. For each plate, 8 μg DNA plus 0.8 ml of serum-free medium is mixed with 48 μl of lipofectamine plus 0.8 ml of serum-free medium, followed by incubation for 30 minutes at room temperature to allow to form the DNA liposome complexes. The serum free medium is added to give a final volume of 6 mL / plate. The cells are rinsed twice with the serum free medium and covered with the DNA liposome mixture. Five hours after transfection, "an equal volume (6 mL) of medium containing 20% FBS (fetal bovine serum) is added." The cells are fed fresh complete medium for 18-24 hours after transfection. They are divided at 72 hours following transfection, at densities of 1:10 and 1:20.The stable transformants are selected by the growth of the cells in the antibiotic containing the medium (G418, Sigma Chemical Company (also known as Geneticin)). The colonies are isolated and tested for the expression receptor by radioligand binding and again measuring the phospho-inositol.

Preparation of Cellular Membrane and Cell Culture. HEK 293 cells stably transfected with human GnRH receptors as described above are grown in MEM (minimal essential medium, available from Sigma Chemical Company) supplemented with 0.1% G418 (Sigma Chemical Company). The cells are homogenized in a buffer A containing 50 mM Tris (pH 74.), 0.32 M sucrose, 2 mM ethylene glycol bis (β-ammoethyl) -N, N, Nr, N '-tetraacetic acid ( EGTA), ~~ 1 mM phenylmethylsulfonyl fluoride (PMSF), 5 μg / ml aprotinen, 5 μg / ml Pepstatin * A, and 1 μg / ml ieupeptin. The homogenized cells are centrifuged at 4 ° C at 20,000 x g for 25 minutes, resuspended in buffer A, and centrifuged again at 4 ° C at 20,000 x g for an additional 25 minutes. The protein of the total membrane is determined with a BCA device (Pierce, Rockford, Illinois). The membranes are stored at -70 ° C at a final membrane protein concentration of 5 mg / ml. Preparation of 125 I-GnRH-A. The analog radioiodinated agonist of GnRH, etiiamide of [des-Gly10, D-Ala9] GnRH (GnRHA), is used as the labeled ligand. One μg of GnRH-A diluted in O.lM acetic acid is added to a borosilicate glass tube coated with Iodogen® (Pierce) containing 35 μL of 0.05M phosphate buffer (pH 7.4-7.6) and 1 mCi of Na ["5I] The reaction mixture is divided and incubated for 1 minute at room temperature, and one minute later the mixture is re-vortexed and allowed to incubate for an additional minute.Two ml of 0.5M acetic acid / 1% BSA is added to the reaction tube, and the mixture is added to a C18 Sep-Pak cartridge.The cartridge is washed with subsequent 5 LH washes, 0 and 5 ml of 0.5M acetic acid, and -then eluted with 5 x 1 ml of 60% CH3CN / 40% acetic acid 0.5M The eluent is diluted with 3 x volume of HPLC buffer A (0.1% TFA in H0) and loaded onto a C18 column. The iodinated product is "eluted at more than 20-25 minutes with a gradient of 25-100% CH3CN containing 0.1% TFA. The radioactive fractions (750 μl / fraction) are collected in clean polypropylene tubes containing 10 μl of 10% BSA. Fractions are evaluated for biological activity by radioligand binding. Active fractions are used to protect the compounds for competitive binding assays. Competitive Radioligand Linkage Assays. Membranes of HEK 293 cells that stably express hGnRHR (human GnRH receptors) are diluted to 0.5-1.0 mg / mL with a test buffer containing 50 mM HEPES (pH 7.4), 1 mM of ethylenediaminetetraacetic acid (EDTA), 2. mM of MgCl_, and 0.1% of BSA. The membranes are incubated with 50,000 CPM (approximately 0.05-0 InM) of 125 I-GnRH-A in a final volume of 200 μi in the presence or absence of competitive agents for 1 hour at room temperature. The tests are stopped by rapid filtration in 96 well GF / C filters soaked in 0.1% polyethylenimine. The filters are washed three times with ice-cold PBS (50 mM NaP0, 0.9% NaCl, 2 mM MgCl, and 0.02% dN NaN i '".4) u i.

Packard of 96 wells. Subsequently, 35 μl of combined scintillation is added to each well of the filter, and the filters are counted with a Packard Top counter. The GnRH link. Sensitive curves are generated for the control dose for GnRH (0.1 nM-100 NM) in each experiment. A dose-sensitive curve is shown in Figure 1, which describes the amount of labeled radio-labeled GnRH limited by cell membranes (CPM) against the concentration of unlabeled GnRH. It will be appreciated by experts that a variety of compounds according to the invention can be prepared. For example, compounds of Formula I having the combination of the radicals X? -X_, Y, and Z shown in Table A below can be prepared, where the radicals Xi-X ^, Y, and Z are identified in the Tables 1, 2, and 3. Table A Exemplary Compounds Table A Exemplary Compound (continued) Table A Exemplary Compound (continued) Table A Exemplary Compounds (Continued) Other Exemplary compounds of Formula I identified in Tables 4 and 5 below were synthesized according to the reaction scheme described above. Crude compounds were tested using the competitive radioligand binding assay described above. The results of the GnRH competitive binding assay are shown in Tables 4 and -5. The radicals X? ~ X2, Y, and Z for the compounds listed in Table 4 are shown in Tables 1, 2, and 3, respectively. The results were obtained for the compounds identified in Table 4 in a single concentration of 10 μM. The specific binding of the diluent controls (vehicle) were determined as the total amount of radioligand binding minus the amount of binding in the presence of lOOnM of GnRH. The percentage of the specific link that remains ("%" in Table 4) in the presence of each compound was calculated according to the following equation:. { Specific link in the presence of the compound / specific link in the presence of the diluent (vehicle)} x 100% In Table 4, a yield of the compound that remains - at 50% could yield a K3 of about 10 μM. Note that the lowest value is for the% link, the highest is the inhibition of the percentage.

Table 4 Results of the Competitive Binding Assay of the GnRH Receptor for the Compound of Formula I Table 4 Results of the Competitive Binding Assay of the GnRH Receptor for the Compound of Formula I (Continued) Table 4 Results of the Competitive Binding Assay of the GnRH Receptor for the Compound of Formula I (Continued) Table 4 Results of the Competitive Binding Assay of the GnRH Receptor for the Compound of Formula I (Continued) Table 4 Results of the Competitive Binding Assay of the GnRH Receptor for the Compound of Formula I (Continued) ki.

I heard i 01 (?? op?: nu? juo3) ¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡ 80 L Table 4 Results of the Competitive Binding Assay of the GnRH Receptor for the Compound of Formula I (Continued) Table 4 Results of the Competitiyo Linkage Assay of the GnRH Receptor for the Compound of Formula I (Continued) fifteen f, Table 4 Results of the GnRH Receptor Competitive Binding Assay for Compounds of Formula I (continued) Table 4 Results of the Competitive Binding Assay of the GnRH Receptor for the Compound of Formula 1 (Continued) Table 5 Results of the Competitive Binding of the GnRH Receptor for the Compound (Each Test at 10 μM) Table 5 Competitive Binding of the GnRH Receptor for the Compound Test Results (Each Test at 10 uM) (Continued) Table 5 Receptor Competitive Binding Assay Results for Compounds (Each Test at 10 μM) (Continued) The invention has been illustrated with reference to the preferred embodiments and exemplary aspects of the invention. Various modifications and adaptations will be clear to the skilled person through the routine practice of the invention in view of the knowledge and developments in the art. Therefore, the invention should be understood to not be limited by the detailed description above, as defined by the appended claims and their equivalents.

It is noted that in relation to this date, the best method known by the applicant to carry out the aforementioned invention, is the conventional one for the manufacture of the objects or products to which it refers. - Having described the invention as above, the content of the following is claimed as property:

Claims (24)

1. CLAIMS 1. A compound of Formula I (I) characterized in that: Z is a group selected from alkyl, alkenyl, alkynyl, cycloalkyl, heterocycle, aryl, heteroaryl, CH_OR, and C (O) OR, substituted or unsubstituted, where R is alkyl, alkenyl, alkynyl, cycloalkyl, aryl, or heteroaryl, substituted or unsubstituted and where the total number of carbon atoms present in Z, optionally does not include substituents, ranging from I to 12; Y is a lipophilic group selected from alkyl, alkenyl, alkynyl, cycloalkyl, aryl, and heteroaryl, substituted or unsubstituted, wherein the total number of carbon atoms - present in Y, optionally does not include substituents, ranging from 6 to 20; Xi is a structural unit, or in space, used to join the functional units CH_NC (0), X_, Y, and Z in the three-dimensional space, which is selected from alkyl, alkenylene, alkynyl, cycloalkyl, aryl, and substituted heteroaryl and unsubstituted such that the count of ~ atoms in the chain portion of the unit linking the central nitrogen (the N atom in Formula I is referred to as "central nitrogen" to avoid confusion with any other substituent produced with nitrogen) for X_ It goes from 3 to 8. Preferably from 4 to 6; and X_ is a basic group qup having a pK- greater than about 8 which is preferably selected tjp g? an id ini] o, ami ini lo, acyllaroidi n i 1, 7i, ln, and am not pu fi tiji n and substituted. ? . A compound, salt, polymer, prodrug, or active metabolite according to claim 1, wherein Z is a furyl, pyrrolyl, naphthyl, thienyl, pyridyl, cycloalkyl of a R carbon aromos, liter d < ^ 1 to R carbon atoms, CH_CFJ; or CF.-. substituted or unsubstituted, R. 3. A compound, sai, multimer, prodrug, or active metabolite according to claim 1, characterized in that Z is furyl, pyrrolyl-phenyl, or CF. 4. A compound, salt, muimeric, prodrug, or active metabolite according to claim 1, characterized in that? is phenyl, naphthyl, substituted or unsubstituted, saturated or partially saturated naphthyl derivative, dibenzyl furyl, saturated or partially saturated dibenzyl ester derivative, quinolinyl, isoquinolinyl, quinoxal inyl, saturated or partially saturated rirloalkyl, or fused polycyclic alkyl. 5. A compound, salt, multimer, prodrug, or active metabolite according to claim 1, characterized in that Y is 4-i? Oppy 1 and i, 4-N, N-dimet i! ami uopropi l xifen lo, 2, 4, -r) -triethoxyphenyl, 2,3-dibenzyloxyphenyl, 2- (4'-rh 1 orofen i 1 x i) fen i 1 or a narcial or totally saturated with them, or 1,2,3,4-tetrahydro-1, 1,4,4,6-pent: amethi Inaftalen-7-yl. 6. A compound, salt, multimer, prodrug, or active metabolite according to claim 1, characterized in that Y is 7. A compound, salt, muitimer, prodrug, or active metabolite according to claim 1, characterized in that - Xi is alkylene, alkylene- (Cs-g) -alkylene, or alkylene- (C = .-?) Aryl. -acykylene. 8. A compound, salt, multimer, prodrug, or active metabolite according to claim 7, characterized in that Xi is selected from the group consisting of: - (CH2) -, - (CH2) 5 ~, 9. A compound, salt, multimer, prodrug, or active metabolite according to claim 1, characterized in that the group X_ has a pK, greater than about 9. 10. A compound, salt, multimer, prodrug, or active metabolite according to claim 1, characterized in that Xi is guanidinyl, amidinyl, or amino, substituted or unsubstituted with one or more lower alkyls. 11- A compound, salt, multimer, prodrug, or active metabolite according to claim 10, characterized in that X_ is guanidinyl. 12. A compound, salt, multimer, prodrug, or active metabolite according to claim 1, characterized in that the compound is of the following formula: or .NH. "? NH 13. A compound, salt, multimer, prodrug, or active metabolite according to claim 1, characterized in that the compound is of the following formula: 14. A compound according to claim 1, characterized in that it is selected from the group consisting of: or a pharmaceutically acceptable salt, multimer, prodrug, or active metabolite thereof. 15. A pharmaceutical composition characterized in that it comprises: a therapeutically effective amount of a GnRH agent selected from the group consisting of the compounds of Formula I and salts, multimers, prodrugs, and pharmaceutically acceptable active metabolites thereof according to claim 1; and a pharmaceutically acceptable carrier or diluent. 16. A pharmaceutical composition according to claim 15, characterized in that the compounds of the Formula I are selected from the compounds of the formula: 17. A pharmaceutical composition according to claim 15, characterized in that the compounds of the Formula I are selected from the "compounds of the formula: 18. A method for regulating the secretion of gonadotropins in mammals, characterized in that it comprises administering a therapeutically effective amount of a GnRH agent selected from the group consisting of the compounds of Formula I and pharmaceutically acceptable multimeric salts, prodrugs, and active metabolites, according to claim 1. 19. A method according to claim 18, characterized in that the compounds of the Formula I are selected from the compounds of the formula: 20. A method according to claim 18, characterized in that the compounds of the Formula I are selected from the compounds of the formula: 21. A compound of one of the following formulas il, III, and IV: = 7 charact eri zed because: j e s 1 or 2; k is 1, 2, 3, 4 or 5; Ru is H or substituted or unsubstituted lower alkyl; RJ is H or a lower alkyl, CN, NO; or CO ^ R1 substituted or unsubstituted; R1"is H or lower alkyl, CH-OR1, (CH2) pOR1, CO_R1, or (CH_) LC (O) R2, where p is an integer from 1 to 6, and R- is H, OR1, SR1 , N (RX) _, or CÍR1.) Substituted or unsubstituted, R11 is H or lower alkyl, CH_0-phenyl, CH_0-benzyl, phenyl, or benzyl, substituted or unsubstituted, or any two of R ", R ?, R1", and R11 taken together form a 5- or 6-membered heterocycle; and wherein each R1 is independently selected from H and lower alkyl, O-lower alkyl, and S-substituted or unsubstituted lower alkyl. 22. A compound according to claim 21, characterized in that the -group is selected from: .NH2 H ^ J '? NH T NBoc 23. A process for developing a compound of Formula I: (i) characterized in that: Z is a group selected from alkyl, alkenyl, alkynyl, cycloalkyl, heterocycle, aryl, heteroaryl, CH_OR, and unsubstituted and substituted C (0) OR, where R is alkyl, alkenyl, alkynyl, cycloalkyl, aryl, or substituted or unsubstituted heteroaryl, and wherein the total number of carbon atoms present in Z, ranging from 1 to 12; Y is a lipophilic group selected from alkyl, alkenyl, alkynyl, cycloalkyl, aryl, and unsubstituted and substituted heteroaryl, where the total number of carbon atoms present in Y, ranging from 6 to 20; Xi is a structural unit containing CHNC (0), Xi, Y, and Z in the three-dimensional space, which is selected from alkyl, alkenyl, alkynyl, cycloalkyl, aryl, and unsubstituted and substituted heteroaryl tai that the atom is in the chain portion of the unit that joins the central nitrogen to X; that goes from 3 to 8; and Xz is a basic group having a pK, greater than about 8, the process is characterized in that it comprises the steps of: (i) reducing the amination in the aldehyde of the formula Y-CHO, where Y is as defined above, with a compound selected from: oc ; And (ii) the acylation of the product of step (i) by reaction with an acyl halide of the formula Z-C0C1, where Z is as defined above. 24. A process according to claim 23, characterized in that it additionally comprises the step of hydrolyzing the product of step (ii) under conditions of acids.