MXPA01005196A - Method of promoting cervical and vaginal secretions - Google Patents

Abstract

The present invention provides a method of stimulating cervical and vaginal secretions in a mammal by treatment with P2Y2, and/or P2Y4 purinergic receptor agonists. Treatment of vaginal dryness associated with menopause, chemotherapy, and various disease states as well as the treatment of vulvar pain is discussed. Suitable purinergic receptor agonists include nucleotide triphosphates such as uridine 5-triphosphate, cytidine 5'-triphosphate and adenosine 5'-triphosphate;dinucleotide polyphosphates such as P1, P4-di(uridine-5') tetraphosphate;and their analogs. These purinergic receptor agonists are useful in stimulating cervical and vaginal secretions and treating vaginal dryness in mammals.

Description

METHOD FOR FAVORING CERVICAL AND VAGINAL SECRETIONS BACKGROUND OF THE INVENTION The mucosa that covers the tracto-reproductive surfaces of women is important in its defense and reproductive function. The mucosal gel, secreted mainly by the endocervical epithelium, offers a barrier to the penetration of sperm and pathogens into the endometrium and a protective covering for the vaginal epithelium. The hydration of the vaginal and cervical mucosa avoids atrophy, provides lubrication during sexual contact, helps the superficial defense against pathogens and modulates the entrance of sperm towards the uterus, etc. (see Gipson I. K. et al., Biology of Reproduction, 60, 58-64 (1999)). The vaginal and ectocervical epithelia are a squamous epithelium (similar to the skin), which does not contain hair cells. The vaginal epithelium is detached during certain phases of the menstrual cycle. The lower endocervical epithelium contains submucosal glands that consist of columnar epithelial cells that secrete mucin, but do not have hair cells. The upper endocervix has the same cells as the lower endocervix and also contains hair cells. The function of the hair cells in the endocervix is not mucociliary clearance. At the time of ejaculation, the sperm are suspended in the vaginal and cervical secretions, which are then actively mixed by the action of ciliary movement and the contraction of the vaginal musculature. This includes movement to and from the cervical mucosal columns so that the sperm can migrate to the mucosa. A function of the hair cells in the endocervix is to expel the non-viable or dead sperm; the cilia move towards the vagina and the viable sperm swim against the gradient. Vaginal dryness is a very common problem that brings physical and emotional discomfort to many women (Key, E., Nurs Stand 5: 24-27 (1991)). This is commonly manifested during sexual contact, which causes dyspareunia and may eventually give rise to apareunia. Although it is traditionally considered to be a condition that affects post-menopausal women, __ can occur during the pre-menopausal and peri-menopausal years. The use of temporary contraceptives can also cause reduction of vaginal moisture in some women (Reginald, W., et al., Br. J. Obstet, Gynecol. 96: 1148-1152 (1989)). Vaginal dryness after childbirth, independent of or as a result of breastfeeding, can be a major complaint (Wisneswki, P., et al., Am. J. Abstet Gynecol. 165: 1249-1254 (1991)). Women who undergo chemotherapy or radiation therapy for malignancies such as leukemia usually experience vaginal dryness as a result of treatment (Cust, M., et al., Br. Med. J. 299: 1494-1497 (1989)). Many disease states such as systemic sclerosis and other systemic autoimmune diseases (Bhadauria, S., et al., Obstet Gynecol. 172: 580-587 (1995)), Ehlersd-Danlos syndrome (Sorokin, Y., et al. , Reprod. Med. 39_: 281-284 (1994)), diabetes mellitus (Strebny, L., et al., Duberes Care 15: 900-904 (1992)), and Sjogren's syndrome (Marchesoni, D., et al., Eur. J. Obstet, Gynecol, Reprod Biol. 63: 49-53 (1995)) report decreased vaginal hydration and lubrication problems as important symptoms associated with the disease. Vulvar pain is defined as excessive sensitivity of the nerves that supply the mucous membrane of the vulva. This permanent burning and sensitivity in the vulvar skin is not caused by infection that can be identified. Nor can it be cured by surgery. Diseases described as "vulvar pain" are also known as vulvodynia / vulvar vestibulitis, vulvitis, vulvar burning syndrome and are often associated with fibromyalgia, irritable bowel syndrome, Sjogren's syndrome, chronic inflammation and Paget's disease as well as in absence of any disease or infection that can be identified. R. Paul St. Armad, MD, an endocrinologist at UCLA, has successfully treated fibromyalgia with uricosuric drugs (gout), especially guaifenesin, a medication used to liquefy mucus (Yount, JJ et al., Omen 's Health Digest 3 (2). 1997). Dr. Armad found that anti-gout medications provide an effective treatment for fibromyalgia, although gout and fibromyalgia are unrelated. Dr. Armad found that 24-hour urine samples taken from patients before and after treatment showed a significant increase in phosphate excretion and a moderate increase in oxalate and calcium after guaifenesin was started. This hypothesis is that an excess of intracellular phosphate, and possibly oxalate, accumulates in the cells of those who suffer fibromyalgia and reduce the formation of energy (ATP) in mitochondria of cells. It should be noted that the role of ATP in Dr. Armad's theory is as a source of energy and not a receptor agonist for P2Y2. Current treatments to increase vaginal moisture are: lubricating agents such as creams or lubricating gels, topical estrogen creams and HRT (hormone replacement therapy). Lubricating gels provide short-term and temporary relief, in view of the fact that these are aqueous preparations that do not contain an active agent for pharmacological use. The effect of estrogen on the maintenance and function of the female genitalia has been well documented. The thickness and folds of the vaginal wall, as well as vaginal lubrication, are dependent on estrogen. Estrogens have also been shown to increase pelvic blood flow in menopausal women and women with surgical or medical oophorectomy. Estrogen deficiency leads to atrophy of the vaginal mucosa and vaginal dryness, which gives rise to symptoms of dyspareunia, sexual dysfunction, as well as a variety of urogenital afflictions. Although estrogen treatment is effective in many women, it may be contraindicated in others for medical reasons such as breast cancer. For example, topical estrogen creams, if used in a regular manner, can be absorbed into the systemic circulation. This can cause stimulation of the endometrium and can cause endometrial hyperplasia and carcinoma (Whirehead, M., et al., N. Eng. J. Med. 305: 1599-1605 (1981)). HRT is effective in relieving symptoms of vaginal atrophy and therefore vaginal dryness but has some contraindications and unwanted risks and side effects. A history of gallbladder disease (N. Eng. J. Med. 290: 15-19 (1974)) or personal or family history of reproductive or breast cancer (Harlap, S., Am. J. Obstet. Gynecol 166: 1986-1992 (1922)) are contraindications for treatment with estrogen. Other contraindications are: history of cerebrovascular accident, cardiovascular disease, deep vein thrombosis, superficial thrombophlebitis, liver disease, smoking, high blood pressure, diabetes, uterine hemorrhage or large fibroids, hyperlipidemia and obesity (Lichtman, R., J. Nurse Mudwifery 36: 30-48 (1991)). An important disadvantage of HRT is the resumption of monthly bleeding due to deprivation, which post-menopausal women will not accept. Some women, although on HRT, still experience a degree of vaginal dryness (Key, E., Nurs, Stand 5: 24-27 (1991)). Uridine 5'-triphosphate (UTP) and polyphosphate dinucleotides such as diuridine tetraphosphate have been shown to be potent agonists of the purinergic receptors for P2Y2 found on the epithelial surface of human respiratory tract. UTP has been found to increase the rate and total amount of mucin secreted by the calciform cells in vi tro (Lethem, M., et al., Am. J. Respir. Cell Mol Biol. 9: 315-322 (1993) ). It has also been shown that UTP increases the secretion of chlorides and, therefore, the secretion of water from the epithelial cells of the respiratory tract in vi tro (Mason, S. et al., Br. J. Pharmacol. 103: 1649-1656 (1991)). Diuridine tetraphosphate has shown beneficial properties in the treatment of different diseases such as chronic obstructive pulmonary disease (COPD). For example, it has been shown that they facilitate the clearance of mucosal secretions from the lungs of an individual such as mammals including humans in need of such treatment for various reasons, including cystic fibrosis, chronic bronchitis, asthma, bronchiectasis, post-operative, pneumonia, primary ciliary dyskinesia (MJ Stwts, III, et al., U.S. Patent No. 5,635,160; PCT International Publication WO 96/40059) and the prevention and treatment of pneumonia in immobilized patients (K.M. Jacobus and H.J. Leighton, US Patent No. 5,763,447). Other therapeutic uses include the treatment of cystic fibrosis, chronic bronchitis, asthma, bronchiectasis, post-operative aletectasia [sic] and Kartagener syndrome (PCT International Publication WO 96/40059), sinusitis (PCT International Publication WO 98/03177) , otitis media (PCT International Publication WO 98/27756), dry eye, retinal detachment, obstruction of the nasolacrimal duct, treatment of female infertility and irradiation due to vaginal dryness through increased mucus secretions and hydration of the epithelial surface, and the improvement of athletes' performance. As a result of the ineffectiveness and risks of current treatments for vaginal dryness, medical researchers have sought to develop alternative treatments. Due to the demonstrated ability of UTP and the polyphosphate dinucleotide, such as diuridine tetraphosphate, to increase the hydration of epithelial secretions of the respiratory tract and stimulate the release of mucins, the applicants were motivated to investigate and the UTP and the dinucleotide polyphosphates can stimulate the hydration and production of mucin in the epithelium and.aginal and cervical.

COMPENDIUM OF THE INVENTION A method for stimulating cervical and vaginal secretions in an individual in need of such treatment is described below. The method of the present invention can be used to augment cervical and vaginal secretions for any reason, including, but not limited to, vaginal dryness treatment and / or treatment of vulvar pain. Vaginal dryness is associated, but not limited to, menopause, delivery, lactation, chemotherapy or radiotherapy, diabetes mellitus, Sjdgren's syndrome, Ehlers-Danlos syndrome, systemic sclerosis and other systemic autoimmune diseases, hysterectomy, urogenital surgery, psychosomatic disorders, anxiety, psychosexual problems and side effects related to pharmacological drugs. The method of the present invention consists of administering a purinergic receptor agonist P2Y2 and / or P2Y: uridine 5'-triphosphate (UTP), cytidine 5'-triphosphate (CTP), adenosine 5'-triphosphate (ATP), P1, P -di (uridine-5'-tetraphosphate or its analogues, in an amount effective to stimulate vaginal and cervical secretions.) Another aspect of the present invention is the JJSO of uridine 5'-triphosphate, cytidine 5'-triphosphate, adenosine 5 '- triphosphate, P1, P4-di (uridine-5 ') tetraphosphate or its analogues for the manufacture of a medicament for carrying out a therapeutic treatment method as mentioned in the foregoing The present invention also discloses pharmaceutical compositions containing uridine 5'-triphosphate, cytidine 5'-triphosphate, adenosine 5'-triphosphate, P1, P-di (uridine-5 ') tetraphosphate or the analogs thereof, with a pharmaceutical carrier for these.

DETAILED DESCRIPTION OF THE INVENTION Applicants have discovered that the 5'-triphosphate nucleotide and the polyphosphate dinucleotides are potent agonists for purinergic receptors found in cervical and vaginal epithelial preparations. The methods of the present invention are an improvement to the currently most used treatments for vaginal dryness, in view of the fact that the compounds of the present invention stimulate the mucin's own production and secretion of the patient as well as the increase in the levels of mucosal hydration, which serve to maintain the protective and lubricating characteristics of the vaginal and cervical mucosa. The methods of the present invention can also be used exclusively, or as an adjuvant for hormone replacement therapy (HRT) or treatment for estrogen replacement (ERT). The present invention provides a method for stimulating cervical and vaginal secretions in a mammal, including a human, in need thereof, by administering the amount of a compound of the formulas I, II, III or IV or a pharmaceutically acceptable ester or salt thereof. effective to increase such secretions. UTP and its analogues are represented in general formula I: Formula I where: Xi, X2 and X3 are each, independent of each other, O "or S ~, preferably X2 and X3 are 0 ~ Ri is O, imido, methylene, or halomethylene, (for example, dichloromethylene or difluorosmethylene); Preferably Ri is oxygen or difluoromethylene, R2 is H or Br, preferably R2 is H, particularly preferably the compounds of formula I are uridine 5'-triphosphate (UTP) and uridine 5'-0- (3 -thiotriphosphate) (UTP? S) A polyphosphate dinucleotide is represented by the general formula II: Formula II wherein: X is oxygen, methylene, dihaluromethylene, imido; n = 0, 1 or 2; m = 0, 1 or 2; n + m = 0, 1, 2, 3 or 4; and B and B 'are each, independent of each other, a purine residue or a pyrimidine residue bound through the 9 or 1 position, respectively; Z = OH or N3; Z '= OH or N3; Y = H or OH; Y '= H or OH; provided that, when Z is N3, Y is H or, when Z 'is N3, Y' is H. The furanose sugar is preferably in the ß configuration. The most preferred furanose sugar is in the ß-D configuration. Preferred compounds of the formula II are the compounds of the formula Formula lia where X = O; n + m = 1 or 2; Z, Z ', Y and Y' = OH; B and B 'are defined in the lie and lid formulas; or X = O; n + m = 3 or 4; Z, Z ', Y and Y' = OH; B = uracil; B 'is defined in the formulas lie and lid; or X = O; n + m = 1 or 2; Z, Y and Z '= OH; Y '= H; B = uracil; B 'is defined in the formulas lie and lid; or X = 0; n + m = 0, 1 or 2; Z and Y = OH; Z '= N3; Y '= H; B = uracil; B '= thymine, or X = 0; n + m = 0, 1 or 2; Z and Z'N3; Y and Y '= H; B and B '= thymine or; X = CH3, CF2 or NH n + m = 1 Z, V, Y and Y '= OH; B and B 'are defined in the formulas lie and lid.

Another group of preferred components of formula II are compounds of formula 11b or pharmaceutically acceptable salts thereof.

Formula IIb wherein: X is oxygen, methylene, difluoromethylene or imido; n = 0 or 1; m = 0 or 1; n + m = 0, 1 or 2; and B and B 'each are independent of each other a purine residue, as in the formula lie or a pyrimidine residue, as in the formula lid, linked through position 9 or 1, respectively. In the case where B and B 'are uracil, bound in position NI to the ribosyl portion, then the total of m + n can equal 3 or 4, when X is oxygen. The ribosyl portions are in the D configuration as shown, but may be in the L, or D and L configuration. The D configuration is preferred.

Formula lie wherein: Ri is hydrogen, Ci-Cs alkyl, phenyl or phenyloxy; wherein at least one hydrogen of the Ci-Cß alkyl, phenyl, phenyloxy, is optionally substituted with a portion selected from the group consisting of: halogen, hydroxy, C?-C4 alkoxy, C?-C 4 alkyl, aryl β-C ?o, carboxy, cyano, nitro, sulfonamido, sulfonate, phosphate, sulfonic acid, amino, C?-C alkylamino, C--C 4 -alkylamino, wherein the alkyl groups are optionally linked to form a heterocycle,? -A (C? -C6 alkyl) CONH (C? -C6 alkyl) -, and? -A (C? -C6 alkyl) NHCO (C? -C6 alkyl) -, where A is amine, mercapto, hydroxy or carboxyl; R2 is 0 or is absent; or Ri and R2 taken together take a fused, 5-membered imidazole ring optionally substituted at the 4 or 5 positions of the ethene portion with C?-C4 alkyl, phenyl or phenyloxy, wherein at least one C de alkyl hydrogen ~ C4, phenyl or phenyloxy is optionally substituted with a portion selected from the group in halogen, hydroxy, C? -C4 alkoxy, C? -C4 alkyl, C6-C? Aryl, C7-C12 arylalkyl, carboxy, cyano, nitro, sulfonamido, sulfonate, phosphate, sulfonic acid, amino, C? -C alkylamino, and C? -C4 di-alkylamino, wherein the dialkyl groups are optionally linked to form a heterocycle; and R3 is hydrogen, amino, C? -C8 alkyl, phenyl or phenyloxy; wherein at least one hydrogen of the amino, C? -C8 alkyl, phenyl or phenyloxy, is optionally substituted with a portion selected from the group consisting of: halogen, hydroxy, C? -C4 alkyl, C0-C aryl? 0, C7-C12 arylalkyl, C?-C4 alkoxy, CC ?2 arylalkyloxy, C?-C 4 alkylthio, phenylthio, C 7 -C 12 arylalkylthio, carboxy, cyano, nitro, sulfonamido, sulfonate, phosphate, sulfonic acid , amino, alkylamino of -C? -C, phenylamino, arylalkylamino of C7-C12, dialkylamino of C? ~ C, wherein the di-alkyl groups are optionally linked to form a heterocycle,? -A (C? C6) CONH (C-C6 alkyl) B-, and? -A (C? -C6 alkyl) 'NHCO (C? -C6 alkyl) B-, where A and B are independently amino, mercapto, hydroxy or carboxyl. Substituted adenine derivatives (formula lie) include 1-adenine oxide, 1, N6 adenine (4 or 5 substituted ethene), adenine substituted at position 6, or aminoadenine substituted at 8, [6-aminohexyl] carbamoyl methyladenine; and amino (hydroxy, thiol and carboxyl) alkyl (C2-C10) acylated adenine in?, wherein the acyl group is selected from, but not limited to, acetyl, trifluoroacetyl, benzoyl, substituted benzoyl, etc., or the carboxylic moiety is present as its ester or amide derivative, for example, ethyl or methyl ester or its methyl, ethyl or benzamido derivative: Formula lid wherein: R4 is hydrogen, hydroxy, mercapto, amino, cyano, C7-C2 arylalkoxy, C? -C6 alkylthio, C? -C6 alkoxy, Ci-C? alkylamino or C? -C4 dialkylamino, in wherein the alkyl groups are optionally linked to form a heterocycle; Rs is hydrogen, acetyl, benzoyl, C? -C6 alkyl, phenyloxy, C1-C5 alkanoyl, aroyl or sulfonate; R d is hydroxy, mercapto, C 1 -C 4 alkoxy, C 7 -C 12 arylalkoxy, C 1 -C 4 alkylthio, amino, disubstituted C 1 -C 5 amino, triazolyl, C 1 -C 6 alkylamino or C 4 -C 4 alkylamino , wherein the dialkyl groups are optionally linked to form a heterocycle or linked to N3 to form a substituted ring; or R5 and R6 taken together form a fused imidazole ring, 5 members between positions 3 and 4 of the pyrimidine ring and form a 3, N4-ethenocytosine derivative, wherein the ethene portion is optionally substituted in the 4 or 5 positions with alkyl of C? -C, phenyl or phenyloxy; wherein at least one C? -C alkyl, phenyl, or phenyloxy hydrogen is optionally substituted with a portion selected from the group consisting of halogen, hydroxy, C? -C4 alkoxy, C? -C alkyl, C6 ~ Co, C7-C2 arylalkyl, carboxy, cyano, nitro, sulfonamido, sulfonate, phosphate, sulfonic acid, amino, C? -C alkylamino and C? -C4 dialkylamino, wherein the dialkyl groups are optionally bound to form a heterocycle. R7 is hydrogen, hydroxy, cyano, nitro or C2-C8 alkenyl, wherein the alkenyl portion is optionally linked through an oxygen to form a ring, wherein at least one hydrogen of the alkenyl portion on the carbon adjacent to the oxygen is optionally substituted with C?-C6 alkyl, phenyl, optionally substituted as described below: substituted C2-C8 alkynyl, halogen, C?-C4 alkyl, CF3, C2-C3 alkenyl, C2-C3 alkynyl , alkylamino, bromo vinyl, ethyl propenoate or propenoic acid; or R and R7 together form a saturated or unsaturated 5- or 6-membered ring linked through N or O in Re, such a ring optionally containing substituents which themselves contain functionalities; provided that when R8 is amino or substituted amino, R7 is hydrogen; Y R8 is hydrogen, amino or C? -C dialkylamino, C? -C alkoxy, C7-C? Arylalkoxy, C? -C4 alkylthio, C7-C? Aralkylthio, carboxamidomethyl, carboxymethyl, methoxy, methylthio , phenoxy or phenylthio, In the general structure of the previous lid formula, dotted lines in positions 2 to 6 are proposed to indicate the presence of single or double bonds in these positions, the relative positions of the double or single bonds being determined by: if the substituents R4, R6 and R7 are capable of keto-enol tautomerism. In the general structures of the above formulas lie and lid, the acyl groups advantageously comprise alkanoyl or aroyl groups. The alkyl groups advantageously contain from 1 to 8 carbon atoms, particularly from 1 to 4 carbon atoms, optionally substituted by one or more suitable substituents, as described below. The aryl groups containing the aryl portions of these groups as aryloxy are preferably phenyl groups optionally substituted by one or more suitable substituents, as described below. The aforementioned alkenyl and alkynyl groups advantageously contain from 2 to 8 carbon atoms, particularly from 2 to 6 carbon atoms, for example, ethenyl or ethynyl, optionally substituted by one or more suitable substituents as described below. Suitable substituents in the aforementioned alkyl, alkenyl, alkynyl and aryl groups are advantageously selected from halogen, hydroxy, C 1 -C 4 alkoxy, C 1 -C 4 alkyl, C 6 -C 12 aryl, C 6 -C arylalkoxy? 2, carboxy, cyano, nitro, sulfonamido, sulfonate, phosphate, sulphonic [sic], amino and substituted amino, wherein the amino is uniquely or doubly substituted by a C? -C4 alkyl, and when? Has double substitution, the groups optionally alkyl being bound to form a heterocycle. For the purpose of further clarifying the above descriptions of the lie and lid formulas, the descriptions can be simplified as follows: R2 is O or is absent; or Ri and R2 taken together can form a fused, 5 membered imidazole ring, optionally substituted; or Rx of group 6-HNR? or R x of the group 8-HNR 3 are selected from the group consisting of: (a) hydrogen, (b) arylalkyl groups (of C x -Cß) with the optionally substituted aryl moiety. (c) alkyl, (d) [6- (A) -hexyl] carbamoylmethyl, where A is independently chosen from NH2, OH, SH and C02H, (e)? -aminoalkyl (from C2-C? 0) (f) )? -hydroxyalkyl (from C2-C? o) (g)? -thiol alkyl (from C2-C? o) (h)? -carboxyalkyl (from C2-C? o) (i) the derivatives? acylates of (e), (f), (g) or (h), wherein the acyl group is acetyl, trifluoroacetyl, benzoyl or benzoylalkyl (of C2-C? 0) substituted, and (j)? -carboxyalkyl (de) C2-C? O) as in (f) above, wherein the carboxyl portion is an ester or an amide.

CTP and its analogues are represented by the general formula II: Formula III where Ri? Xi X2 and X3 are as defined in formula I; R5 and R6 are H, while R7 is absent and there is a double bond between N3 and C4 (cytosine); or R5, R6 and R7 taken together with -CH = CH-, forming a ring from N3 to N4 with a double bond between N4 and C4 (3, N4-ethenocytosine) optionally substituted at the 4 or 5 position of the ethylene ring.

ATP and its analogues are represented by general formula IV: Formula IV wherein Rx, Xx, X2 and X3 are defined as in formula I; R3 and R are H, while R2 is nothing [sic] and there is a double bond between NI and C6 (adenine); or R3 and R are H, while R2 is O and there is a double bond between NI and C6 (1-adenine oxide), or R3, R4 and R2 taken together are -CH = CH-, forming a ring from N6 to NI with a double bond between N6 and C6 (LN6-ethenoadenine). For simplicity, the formulas I, II, III and IV herein illustrate the active compounds in the D configuration that occurs in nature, but the present invention also comprises the compounds in the L configuration and mixtures of the compounds in the D configurations. and L, unless otherwise specified. The D configuration that occurs in nature is preferred.

The compounds of the invention may be present in the form of their pharmaceutically acceptable salts such as, but not limited to, alkali metal salt such as sodium or potassium; alkaline earth metal salt such as manganese, magnesium or calcium; or an ammonium or tetraalkylammonium salt, that is, NX4 + (where X is C? _4). Salts acceptable for pharmaceutical use are salts which retain their desired biological activity of the parent compound and do not confer undesirable toxicological effects. The compounds of the invention are also present in the form of the prodrugs, usually containing ester or amide moieties in the heterocyclic and furanosyl hydroxyl of the compound. Another aspect of the present invention is a method of treating a mammal with vaginal dryness arising from, but not limited to, menopause, parturition, lactation, chemotherapy or radiotherapy, diabetes mellitus, Sjdgren's syndrome, Ehlers-Danlos syndrome, systemic sclerosis or other systemic autoimmune diseases, hysterectomy, urogenital surgery, psychosomatic disorders, anxiety, psychosexual problems and collateral effects related to pharmacological drugs. It is also contemplated that the method of the present invention can be used to increase vaginal moisture and lubrication in healthy women for purposes of facilitating sexual intercourse. It is further contemplated that the method of the present invention would be particularly useful for a woman who has as sexual partner an individual who is undergoing treatment with Viagra® or another of these drugs used for the treatment of erectile dysfunction. The present invention further provides pharmaceutical compositions comprising a dosage form containing the purinergic receptor agonists for P2Y2 and / or P2Y4 selected from the group consisting of general formula I, for example, uridine 5'-triphosphate (UTP) and its analogues; general formula II, P1, P4-di (uridine) -5 '-tetraphosphate (U2P4) and its analogs, general formula III, for example, cytidine 5'-triphosphate (CTP) and its analogs, and general formula IV, for example, adenosine 5'-triphosphate (ATP) its analogues. The compounds described herein can be administered to the cervical and / or vaginal mucosa of a patient by any convenient means, but administration by a solution, gel, suspension, cream, foam, pessary or tablet containing the active compound is preferred. . Otherwise, the active compounds can be administered by continuous release of a vaginal ring (Stump, P., Obstet, Gynecol 75: 98 (1990)) or an intrauterine device (Anderson, K. et al., 79: 963 (1992)). The topical solution, gel, jelly, ointment, cream, foam, pessary, or tablet contains the active compound in a physiologically compatible vehicle, as those skilled in the art of topical gynecological delivery systems can select using the traditional criteria. Solutions formulated for administration to the vagina are commonly known as irrigations. These are sterile solutions prepared in a common form of sterile injections that are proposed for and prepared as a sterile single-use solution. The gel (water-soluble bases) and creams (bases that can be removed with water) can be developed depending on the characteristics of the transport of the asset. For gel formation, gelling agents (eg, polyethylene glycols, gelatin, tragacanth and cellulose derivatives) and preservatives (antimicrobial agents) can be used. It is possible to add stabilizers, antioxidants and buffer solutions as needed. For example, a convenient gel formulation in this invention includes the active compound, glycerin, a cellulose derivative (e.g., hydroxyethylcellulose), a preservative (e.g., methyl paraffin), and water. For cream formulations it is possible to develop bases for emulsion. Petrolatum and stearyl alcohol can be used in the oil phase. Petrolatum contributes to the ability to retain water from the formulation, and the silyl alcohol can serve as an emulsifier. The aqueous phase of a base for emulsion may contain water-soluble components of the emulsion system, preservatives, stabilizer, anti-oxidant, buffer and emulsifier. It is also possible to use glycerin, propylene glycol or polyethylene glycol to minimize the loss of water in the finished product. Ointments are semi-solid preparations consisting of the active ingredient incorporated in a fatty, waxy or synthetic base. Examples of suitable creams include, but are not limited to, water-in-oil emulsions, and oil-in-water emulsions. Water-in-oil creams can be formulated using a suitable emulsifying agent with similar, but not limited, properties to those of DS fatty alcohols such as cetyl alcohol or cetostearyl alcohol and emulsifying wax. Oil-in-water creams can be formulated using an emulsifying agent such as cetomacrogol emulsifying wax. Suitable properties include the ability to modify the viscosity of the emulsion and physical and chemical stability over a wide pH range. The base for water soluble or miscible cream may contain a preservative system and may also be buffered to maintain an acceptable physiological pH. The foam preparations can be formulated to be supplied from a can for pressurized aerosol, by means of a suitable applicator, using inert propellants. Suitable excipients for foam base formulation include, but are not limited to, propylene glycol, emulsifying wax, cetyl alcohol and glyceryl stearate. Potential preservers include methylparaben and propylparaben. Pessaries are solid forms for unit dosage specially formed for insertion into the vagina and may be composed of a base that melts at body temperature or dissolves when in contact with mucosal secretions. Examples of suitable bases include, but are not limited to, theobroma oil, synthetic fatty bases (e.g., Witepsol), polyethylene glycols (macrogols), and glycerol bases for suppositories. Vaginal tablets are composed of the active ingredient contained within a solid base dosage form which may include, but is not limited to, excipients such as lactose, microcrystalline cellulose, corn starch, magnesium stearate, silicon dioxide, and hydroxypropylmethylcellulose. In addition to the topical administration method described above, there are some methods of administering the compounds of the present invention systemically. Such a medium would involve an aerosol suspension of respirable particles comprising the active compound, which is inhaled by the individual. The active compound would be absorbed into the bloodstream by the lungs and would make contact with the cervical and / or vaginal tissues in an effective amount for pharmaceutical use. The respirable particles can be liquid or solid, with a particle size small enough to pass through the mouth and larynx with inhalation; in general, the particles would have an interval of from about 1 to 10 microns, but more preferably 1-5 microns in size are considered respirable. Other means of systemically administering the active compounds would include oral administration, in which the pharmaceutical compositions containing the compounds of the formulas I, II, III or IV are in the form of tablets, dragees, aqueous or oily suspensions, powders or granules. dispersible, emulsions, hard or soft capsules, syrups, elixirs or transdermal delivery devices. Compositions proposed for oral use may be prepared according to any method known in the art for the manufacture of pharmaceutical compositions and these compositions may contain one or more agents selected from the group consisting of: sweetening agents, flavoring agents, coloring agents and agents preservatives in order to provide elegant and pleasant pharmaceutical preparations. The tablets contain the active ingredient in admixture with pharmaceutically acceptable, non-toxic excipients which are suitable for the manufacture of tablets. These excipients can be, for example, inert diluents such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents, for example corn starch or alginic acid, binding agents, for example, starch, gelatin or acacia; and lubricating agents, for example magnesium stearate, stearic acid or talc. The tablets may be uncovered or they may be covered by known techniques to delay disintegration and absorption in the gastrointestinal tract and thus cause a prolonged action for a longer time. For example, it is possible to employ a material to delay time such as glyceryl monostearate or glyceryl distearate. Formulations for oral use can also be presented as hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate or kaolin, or as soft gelatin capsules wherein the ingredient active is mixed with water or an oily medium, for example, peanut oil, liquid paraffin or olive oil. The means of further systemic administration of the active compound to the cervical and vaginal tissues of the individual would include a suppository form of the active compound, such that an effective amount for therapeutic use of the compound reaches the cervical and vaginal tissues by absorption and systemic circulation. .

The aforementioned administration means can be applied during or after surgical procedures, for example, by intra-operative and post-operative [sic] installation. The amount of the active compound included in the pharmaceutical composition is an amount sufficient to reach the concentrations of the active compound in the cervical and / or vaginal mucosa of the individual from about 10"7 to about 10 ~ 3 moles / liter, and more preferably from about 10 ~ 6 to about 10"1 moles / liter. Depending on the solubility of the particular formulation of the active compound that is administered, the daily dose to favor the production of mucin and / or cervical and / or vaginal hydration can be divided between one or several unit dose administrations. The total daily dose for UTP (e.g., may be in the range of 1 to 1000 milligrams, depending on the age and condition of the individual, provided in a scheme up to 4 times a day or as necessary to cope with exacerbations Some compounds of formulas I, II, III and IV can be prepared by methods well known to those skilled in the art and according to known procedures (Zamecnik, P. C, et al., - Proc. Nati. Acad Sci. USA 89: 2370-2373 (1992), Ng, K. et al., Nucleic Acids, Res. 15: 3572-3580 (1998), Jacobus, KM et al., U.S. Patent No. 5,789,391; and Pendergast, W., et al., U.S. Patent No. 5,837,861, and International Patent Application WO 98/34942)). Some are commercially available, for example, from Sigma Chemical Company, PO Box 14508, St. Louis, MO 63178. The synthetic methods of U.S. Patent No. 5,789,391 and of International Patent Application WO 98/34942 are incorporated herein by reference. they are incorporated herein by reference.

EXAMPLES Example 1: In vitro short-circuit (Isc) measurements The UTP compound is a potent agonist of the P2Y2 and P2Y4 purinergic receptors in cervical and vaginal tissue preparations by in vitro evaluation by administering UTP to sufficient tissue culture to reach UTP concentrations in the mucosa from about 10 ~ 7 to about 10_1 moles / liter. (Rojanasakul, Y., et al., Pharm. Res. 9_ 1029-34 (1992)); Bachgaard, E., et al., Int. J. Pharm. 106: 237-242 (1994); Gipson, I., et al., Biol. Reprod. 56: 999-1011, (1997)). Specifically, New Zealand albino rabbits, white, female, ovariectomized are sacrificed and removed from the vaginal tissue. The fabric is mounted on a support ring and fastened in a Ussing chamber. The Isc is measured as it flows from the epithelial side to the serous side of the tissue. Approximately half of this current corresponds to the movement of chlorides through the membrane and therefore, this is an accurate measure of the movement of the corresponding fluid.

Example 2: Cellular localization of gene expression of the receptor for nucleotide P2Y2 in endocervical and vaginal epithelial tissues of monkey by non-isotopic in situ hybridization Fabrics The tissues for the study were obtained from the Tulane Regional Primate Research Center (Covington, LA). The tissues included in this study were vagina and cervix. The tissues were removed from the 3.25-year-old Indian rhesus monkey immediately after death and instantaneous freezing in OCT medium. The frozen tissues were shipped overnight in dry ice and stored at -80 ° C before cryosection. The tissues were cut into sections of 5 microns and mounted on microscope slides for staining with hematoxylin and eosin (H &E) staining with. alciano blue / periodic acid displacement (AB / PAS), and hybridization in si tu (ISH).

Evaluation of tissue sections. Tissue sections stained with H & E were prepared to assess the amount and orientation of the tissues in the study. Examination of the slides with H & And he indicated that all tissues were suitable for ISH.

Synthesis of ribonsonda. A PCR product containing 253-651 nucleotides of a human P2Y2-R cDNA was prepared. Nucleotides 272-627 of P2Y2-R were reamplified with PCR primers designed to eliminate flanking plasmid sequences and incorporate a T3 upstream promoter or a downstream T7 promoter. The resulting PCR products were used to synthesize riboprobes labeled with digoxigenin by in vitro transcription (IVT). The antisense and sense riboprobes were synthesized using T7 and T3 RNA polymerases, respectively, in the presence of digoxigenin-11-UTP (Boehringer-Mannheim) using a MEGAscrip IVT (Ambion) equipment according to the manufacturer. After IVT, the template DNA was degraded with DNase-1 and the unincorporated digoxigenin was removed by ultrafiltration. The integrity of the riboprobe was evaluated by electrophoresis through a denaturing polyacrylamide gel. The apparent molecular size was estimated by comparison with the electrophoretic mobility of an RNA ladder of 100-1000 base pairs (Ambion). The performance and labeling of the probe was evaluated by immunochemical blot. The riboprobes were dosed in 5 μl aliquots and stored at -80 ° C until used for ISH.

Hybridization ip situ. The frozen tissues were cut into sections of 5 microns, mounted on SuperFrost Plus slides (Fisher Scientific), and then fixed for 15 minutes in 4% formaldehyde in PBS at pH 7.4. The sections were pre-hybridized in the absence of the probe, then incubated overnight in buffer for hybridization containing 400 ng / ml of the antisense or sense probe. After hybridization, the slides were subjected to a series of rigorous post-hybridization washes to reduce non-specific staining. Hybridization was visualized by immunohistochemistry using anti-digoxigenin Fab conjugated with alkaline phosphatase and tetrazolium chloride nitro blue [sic] -bromo-chloro indolyl phosphate (Boheringer-Mannheim) according to the manufacturer. The tissue sections were counterstained with nuclear fast red. Negative controls included tissues of the cervix and vagina stained with the P2Y2-R sense probe.

Results / Both endocervical and vaginal epithelial tissues show positive staining using the antisense probe, but negative staining using the sense control probe. The results demonstrate that the endocervical epithelial cells (including the calciform cells) and the stratified squamous epithelium lining the vaginal canal contain the message for the P2Y2 receptors. The results support the reasoning of the present invention, that the activation of the receptors P2Y2 found in the vaginal and cervical epithelium will stimulate vaginal and cervical secretions in mammals.

Example 3. Effects of P2Y2 agonists on the health of the vaginal mucosa in oophorectomized rabbits The purpose of this experiment is to investigate the effect of U2P4 and dCP4U on the health of the vaginal mucosa as assessed by clinical and pathological indicators in a female animal model under conditions of estrogen deprivation and estrogen replacement.

Evaluations Daily evaluations. The health of the mucosa is evaluated by determining the vaginal pH, vaginal lubrication and vaginal smear according to Hubbard et al., (Lab. Anim. Sci. 47: 36-39 (1997)); Bachman et al., (Clinical Sexual Practice 8: 11-17 (1992)) at the same time each day one hour after dosing. Initial vaginal pH measurements are recorded using a digital pH meter (Sandhill Scientific). The lubricacicr. vaginal is evaluated using specially designed tampons, previously weighed, and vaginal smears by fixation, staining and visual examination by pat: logos.

Periodic evaluations. An objective measure of vaginal lubrication is determined by inserting a previously weighed tampon and recording the difference in weight over time. A vaginal smear for cytology (vacuolation, basal, parabasal and superficial cells) is performed at the same time on day 0 (initial data), 4, 9 and 14.1 hours after dosing. The vaginal smear of the baseline is performed, the visual examination of the external genitalia. The vaginal smears are fi xed; n, stain with pap smear. A pathologist classifies by number and evaluates the vacuolation and percentages of the basal, parabasal and superficial cells.

Printing cytology. The animals of each group are then sacrificed using pentobarbitol (8 ml IV). Only after sacrifice is cytology performed by endocervix impression to evaluate the density of calciform cells. The vagina in its integrity (upper and lower segment) as well as the cervix is removed and opened lengthwise). The impression cytology of the vaginal and cervical samples is collected with strips of polyvinylidene difluoride from the vaginal wall and the endocervical surface for evaluation of mucin by PAS staining. The vaginal samples are fixed in 10% neutral buffer with formalin, and then embedded in paraffin, cut at 5 microns and stained with hematoxylin and eosin. Blind subjective classification is performed on vaginal samples using a vaginal atrophy index (Hubbard, 1997, Bachman, 1992). The data is collected and analyzed by the appropriate statistical methods to find out the importance.

Experimental protocol _ To evaluate the effect of P1, P4-di (uridine 5 ') - tetraphosphate (U2P4) and [P1- [5' - (2 '- (deoxycytidine)] P4- (5' - (uridine)] - tetraphosphate (dCP4U) in the density of vaginal calciform cells in oophorectomized animals with and without estrogen replacement therapy, the following experimental protocol will be performed.

Ovariectomy and estrogen replacement treatment in animals. White female rabbits, New Zealand (4 kg) are divided into 6 groups of 6 animals per group. (See Table 1). Group 1 remains intact and without treatment (without surgery, without placebo, without treatment with medication); This is used as a reference to obtain baseline information. The remaining groups of animals (groups 2-6) are bilaterally ovariectomized with anesthesia, as described (Hansen et al., Am. J. Obstet, Gynecol., 175: 1272-1280 (1996), Zandberg et al., Arterioscler. Thromb. Vasc. Biol. 18: 1844-1854 (1998) Two weeks after ovariectomy, group 2 does not receive treatment, group 3 receives placebo, group 4 receives P1, P4-di (uridin 5 ') - tetraphosphate (U2P4) and group 5 receives (P1 [5 '- (2'- (deoxycytidine)] P4- (5' - (uridine)) -tetraphosphate (dCP4U) Group 6 receives an implant with Azat pump supplying estradiol (200 μg / day, for 14 days) Group 6 is further subdivided into 3 subgroups, group 6A receives placebo, group 6B receives U2P, and group 6C receives dCP4U.The concentration of estrogen in the blood of each animal is determined by radioimmunoassay, pre-oophorectomy, 14 days after oophorectomy, and 14 days after estrogen replacement therapy Table 1 summarizes the experimental protocol ntal.

Table 1. Experimental protocol to study the effects of P2Y2 agonists on the health of the vaginal mucosa in oophoreticized rabbits Eight study groups of 6 rabbits per group = 48 animals minimum.

The invention, and the form and process of preparing and using it, will now be described in complete, clear, concise and accurate terms to enable any expert in the art to which it belongs, to prepare and use it. It should be understood that the foregoing describes the preferred embodiments of the present invention and that the modification may be made therein without departing from the spirit or scope of the present invention as set forth in the clauses.

Claims (10)

1. A method for stimulating cervical and vaginal secretions in a mammal in need thereof, by administering an effective amount for stimulating the secretion, of a compound of the formulas I, II, III or IV: Formula I wherein: Xi, X2 and X3 are each, independent of each other, 0"or SJ Rx is 0, imido, methylene, or halomethylene; R2 is H or Br, preferably R2 is H; Formula II wherein: X is oxygen, methylene, dihaluromethylene, imido; n = 0, 1 or 2; m = 0, 1 or 2; n + m = 0, 1, 2, 3 or 4; and B and B 'are each, independent of each other, a purine residue or a pyrimidine residue linked through position 9 or 1, respectively; Z = OH or N3; Z '= OH or N3; Y = H or OH; Y '= H or OH; provided that, when Z is N3, Y is H or, when Z 'is N3 Y' is H; or Formula III wherein Ri / i / X2 and 3 are as defined in formula I; R5 and Re are H, while R7 is absent and there is a double bond between N3 and C4 (cytosine); or R5, Re and R taken together with -CH = CH-, forming a ring from N3 to N4 with a double bond between N4 and C4 (3, N4-ethenocytosine) optionally substituted at the 4 or 5 position of the ethylene ring; or Formula IV wherein Ri, Xi, X2 and X3 are defined as in formula I; R3 and R4 are H, while R2 is absent and-there is a double bond between NI and C6; or R3 and R are H, while R2 is 0 and there is a double bond between NI and C6, or R3, R4 and R2 taken together are -CH = CH-, forming a ring from N6 to Ni with a double bond between N6 and C6; or the esters or salts acceptable for pharmaceutical use thereof.

2. The method of claim 1, wherein the compounds of the formula II are those of the formula lia: Formula bundle where X = O; n + m = 1 or 2; Z, Z ', Y and Y' = OH; B and B 'are defined in the lie and lid formulas; or X = O; n + m = 3 or 4; Z, Z ', Y and Y' = OH; B = uracil; B 'is defined in the formulas lie and lid; or X = O; n + m 1 or 2; Z, Y and Z '= OH; Y '= H; B = uracil; B 'is defined in the formulas líe and lid; or X = 0; n + m = 0, 1 or 2; Z and Y = OH; Z '= N3; Y '= H; B = uracil; B '= thymine, or X = 0; n + m = 0, 1 OR 2; Z and Z'N3; Y and Y '= H; B and B '= thymine or; _ X = CH3, CF2 or NH n + m = 1 Z, V, Y and Y '= OH; B and B 'are defined in the formulas lie and lid. Formula líe wherein: Ri is hydrogen, C? -C8 alkyl, phenyl or phenyloxy; wherein at least one hydrogen of C?-C8 alkyl, phenyl, phenyloxy, is optionally substituted with a portion selected from the group consisting of: halogen, hydroxy, C?-C4 alkoxy, C?-C4 alkyl, aryl of C6-C? o, carboxy, cyano, nitro, sulfonamido, sulfonate, phosphate, sulphonic acid, amino, C1-C4 alkylamino, C? ~ C4 di-alkylamino, wherein the alkyl groups are optionally linked to form a heterocycle,? -A (C? -C6 alkyl) CONH (C? -C6 alkyl) -, and? -A (C? -C6 alkyl) NHCO (C? -C6 alkyl) -, where A is -amino, mercapto, hydroxy or carboxyl; R2 is O or is absent; or Ri and R2 taken together take a fused, 5-membered imidazole ring optionally substituted at the 4 or 5 positions of the ethene portion with C? ~C alkyl, phenyl or phenyloxy, wherein at least one C de alkyl hydrogen -C 4, phenyl or phenyloxy is optionally substituted with a portion selected from the group in halogen, hydroxy, C 1 -C 4 alkoxy, C 1 -C 4 alkyl, C 6 -C 6 aryl, or C 7 -C 2 arylalkyl, carboxy, cyano, nitro, sulfonamido, sulfonate, phosphate, sulfonic acid, amino, C? -C4 alkylamino, and C? -C-dialkylamino, wherein the dialkyl groups are optionally linked to form a heterocycle; and R3 is hydrogen, amino, C? -C8 alkyl, phenyl or phenyloxy; wherein at least one amino hydrogen, C? -C8 alkyl, phenyl or phenyloxy, is optionally substituted with a portion selected from the group consisting of: halogen, hydroxy, C? -C4 alkyl, Co-Cio aryl, C 1 -C 2 arylalkyl, C 1 -C alkoxy, CC 2 arylalkyloxy, C 1 -C 4 alkylthio, phenylthio, C 7 -C 2 arylalkylthio, carboxy, cyano, nitro, sulfonamido, sulfonate, phosphate, sulfonic acid, amino, C? -C alkylamino, phenylamino, C7-C12 arylalkylamino, C? -C4 dialkylamino, wherein the dialkyl groups are optionally linked to form a heterocycle,? -A (C-C6 alkyl) CONH (C -C6 alkyl) B-, and? -A (C? -C6 alkyl) NHCO (C? -C6 alkyl) B-, where A and B are independently amino, mercapto, hydroxy or carboxyl. Formula lid wherein: R4 is hydrogen, hydroxy, mercapto, amino, cyano, aralkoxy [sic], C-C6 alkylthio, C-Cd alkoxy, C? -Ce alkylamino or dialkylamino [sic], wherein the alkyl groups of the dialkylamino are optionally linked to form the heterocycle; Rs is hydrogen, acyl, C 1 -C 6 alkyl, aroyl, C 1 -C 5 alkanoyl, benzoyl or sulfonate; R6 is hydroxy, mercapto, alkoxy, aralkoxy, Ci-C5 disubstituted Ci-Cß amino alkyl, triazolyl, alkylamino or alkylamino alkylthio, wherein dialkylamino alkyl groups are optionally linked to form a heterocycle or linked to N3 to optionally form a ring replaced; or R5-R6 together form a saturated or unsaturated ring of 5 or 6 members linked through N u 0 in R 6, wherein the ring is optionally substituted; R7 is selected from the group consisting of: a) hydrogen b) hydroxy c) cyano d) nitro e) alkenyl, wherein the alkenyl portion is optionally bonded through oxygen to form a ring optionally substituted with alkyl or aryl groups on the carbon adjacent to oxygen f) substituted alkynyl g) halogen h) alkyl i) substituted alkyl j) CF3 k) C2-C3 alkenyl, m) substituted ethenyl n) C-C3 alkynyl, oo) substituted alkynyl; R8 is selected from the group consisting of: a) Hydrogen _ b) Alkoxy c) Arylalkoxy d) Alkylthio e) Arylalkylthio f) Carboxamidomethyl g) Carboxymethyl h) Methoxy i) Methyl j) Phenoxy k) Phenylthio 1) Amino m) Alkylamino, and n) Dialkylamino

3. The method of claim 1, wherein the compounds of the formula II are those of the formula Ilb: Formula IIb wherein: X is oxygen, methylene, difluoromethylene or imido; n = 0 or 1; m = 0 or 1; n + m = 0, 1 or 2; and B and B 'each are independent of each other a purine residue, as in the formula lie as described in clause 2, or a pyrimidine residue, as in the formula lid, as described in clause 2, linked to through position 9 or 1, respectively provided that when B and B 'are uracil, linked at the position Nl to the ribosyl portion, then the total of m + n is equal to 3 or 4, when X is oxygen.

4. The method of claim 1, wherein R2 of formula I is H.

5. The method of claim 1, wherein the furan sugar of the formula II is in the ß-D configuration.

6. A method of treating a mammal with vaginal dryness by administering an effective amount for the treatment of vaginal dryness of a compound of formulas I, II, III or IV as described in clauses 1-5.

7. A pharmaceutical composition containing a compound of the formulas I, II, III or IV as described in clauses 1-5 together with a pharmaceutically acceptable carrier for it in the form of a liquid suspension or gel.

8. The method of claim 6, wherein the amount of the compound of the formulas I, II, III or IV administered to the mammal is sufficient to achieve a concentration in the cervical and / or vaginal mucosa from about 10"7 moles / liter to about 10"1 moles / liter.

9. The method of claim 6, wherein the amount of the compound of the formulas I, II, III or IV administered to the mammal is sufficient to achieve a daily dose of between 1 to 1000 milligrams.

10. A method of treating a mammal with vulvar pain by administering an amount effective for the treatment of vulvar pain of a compound of formulas I, II, III or IV as described in clauses 1-5.