HK1022099A - Method of treating ciliary dyskinesia with uridine triphosphates and related compounds - Google Patents

Description

Methods for treating ciliary dyskinesia with uridine triphosphates and related compounds are described

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Technical Field

The present invention relates to stimulating ciliary beat frequency to promote mucociliary or cough clearance of mucus secretions remaining in the lungs, sinuses or ears of a patient by administering specific uridine, adenosine triphosphate or cytidine triphosphate.

Background

Mucociliary clearance is an important defense mechanism for the human trachea and middle/inner ear canal. The coordinated beating of cilia in the nose, trachea, bronchi and middle ear propels the mucosal layer and its entrained microorganisms and other particles trapped in the mucosa for endopharyngeal movement. The normal function of the system depends on the frequency and coordination of ciliary beating and the nature of its mucosa. The mucociliary clearance system consists of three components: (1) a mucin layer formed by secretion of mucin by goblet cells, (2) cilia which transport excess mucin layer by synchronous beating, and (3) a liquid layer at the edge of cilia which surrounds the cilia and is less viscous than the mucin layer, which allows free movement of the cilia. The electrolyte and water concentrations of the cilial border layer are regulated by the epithelial cells of the lumen. (R.Boucher et al, adenosine and adenine nucleotides: from molecular biology to integrated physiology, pages 525-32, entitled "mechanism and therapeutic action of uridine triphosphates in the lung" (L.Belladielli et al, eds., Alumwer Academic Publishers,1995, Boston, USA).

Primary Ciliary Dyskinesia (PCD) is a progressive disease characterized by defects in the superstructure of cilia and dyskinesias resulting in the absence of ciliary movement or ciliary movement disorders. The most common clinical symptoms of PCD are chronic respiratory diseases (e.g., sinusitis, rhinitis, and bronchitis) and otitis media. Since PCD patients do not have mucociliary clearance (MCC) or mucociliary clearance (MCC) is severely impaired, the only clearance mechanism or mechanism of secretion removal is cough. Cough clearance can be measured in the same way as for the MCC described above. PCD also impairs the propulsion of sperm, resulting in male infertility. (D.Schidlow, Ann.Alergy 73(b),457-68 (1995)). PCD also causes impairment of the motility of certain cells of the immune system, including nucleophilic cells and macrophage cells. PCD and may be the primary cause of hydrocephalus formation due to ciliary dysfunction. (M.Greenstone, Arch Dis Child 59,481-82 (1984)). It was calculated that PCD occurred in one of 16,000 birth lives, and it was estimated that 50% of patients also had a position flip (right heart). The three-in-one syndrome of bronchitis, sinusitis and inversion of position (right heart) is known as catagen (Kartagener) syndrome. (M.Sleigh, Lancet ii,476 (1981)). It is now assumed that Kartagener's syndrome is due to lack of ciliary movement of the embryo, resulting in random rotation of the gut with a positional reversal (right heart) in half of the cases and a normal heart position in the other half. (B.Afzelius Science 193,317-19 (1976)). The clinical course of PCD is primarily characterized by sinus and ear infections early in life, with progressive changes to lung/lower respiratory tract disease after adulthood. Chronic respiratory infections result in chronic obstructive changes in lung tissue, progressive loss of lung function, and ultimately death.

A secondary and more common form of ciliary dyskinesia is the acquired form of the disease. Chronic inflammation caused by severe viral or bacterial respiratory infections, long-term smoking, severe air pollution, chemical or thermal damage to the trachea, intubation and mechanical ventilation, and near drowning all cause alterations in the cilia structure, including rupture of cell membranes, loss or coalescence of microtubules, and formation of complex cilia, all of which result in abnormal or abolished cilia function (j. balllenger Ann Otol Rhinol Laryngol 97(3pt.1),253-58 (1988); m. pedersen Lung 168suppl, 368-76 (1990)). Respiratory infections that lead to secondary ciliary dyskinesia include colds, adult respiratory distress syndrome, and Ventilator Associated Pneumonia (VAP) in Intensive Care Unit (ICU) patients. Some cases of acquired ciliary dyskinesia may be reversed with appropriate and timely treatment; however, permanent damage and/or continued exposure to the above factors may irreversibly damage the cilia. For chronic lung infections, progressive loss of lung function and obstructive lung disease, the clinical symptoms and progression may be very similar to PCD.

Typical mammalian respiratory epithelial cells contain about 200 cilia. Each cilium has 9 peripheral microtubule pairs and two central tubes. Each peripheral pair contains an a subunit and a B subunit, each a subunit having a set of flexure arms attached to it, the so-called internal and external motor protein (dynein) arms. These motor protein arms contain atpases, enzymes that break down Adenosine Triphosphate (ATP), providing energy for ciliary movement. Since the most common suprastructure abnormality associated with primary ciliary dyskinesia is the general lack of moving arms (b.afzelius et al, journal of cell biology (j.cell Biol)66,225-32(1975)), researchers began investigating whether extracellular use of ATP and atpase could activate immobile cilia in vitro. (J.Forrest et al, Am Rev RespDispis 120,511-15 (1979)). Although the result was positiveHowever, this finding was not reproduced by others. Later discovered that Ca was used extracellularly²⁺And cAMP increase the beating frequency of respiratory cilia (A. Lansley et al, journal of physiology in the United states (am. J. physiol)263, L232-42 (1992)). It has not been established that any therapy stimulates ciliary beat in the case of complete ciliary immobility. In such cases, a beneficial therapy is to increase the hydration of the viscous mucosal secretions.

Applicants have now found that extracellular nucleoside triphosphates, particularly uridine 5' -triphosphate, regulate three components of the ciliary transport system. UTP stimulates ciliary beat frequency (r. boucher et al, supra); UTP stimulates goblet cells to secrete mucin (M.Lethem et al, Am J Respir Cell Mol Biol 9,315-22 (1993)); UTP stimulation of Cl in tracheal epithelial cells^-Increases the hydration of the peripheral ciliary liquid layer (M.Knowles et al, N Eng J. Med 325,533-38 (1991)). Applicants have shown that UTP is safe and promotes cough clearance in PCD patients. (P.Noone et al, filed in the Abstract of the American thoracic society International conference on communication 1996).

In summary, the various clinical symptoms of ciliary dyskinesia, such as absence of mucociliary clearance or impaired mucociliary clearance in the respiratory tract and middle ear/inner ear canal, impaired sperm propulsion and impaired motility of nucleophilic cells and macrophages, may be determined by administering UTP and its related compounds and other nucleotide phosphates, such as adenosine 5' -triphosphate (ATP); cytidine 5' -triphosphate (CTP); 1, N₆-ethano adenosine triphosphate; adenosine 1-oxide triphosphate; 3, N⁴-ethanocytidine triphosphate; p₁,P₄-bis (adenosine-5') tetraphosphate (A)₂P₄) (ii) a Or P₁,P₄-di (uridine-5') tetraphosphate (U)₂P₄) May be improved or reduced.

Summary of the invention

A method for treating ciliary dyskinesia in a subject in need thereof is disclosed. The method comprises administering to the patient a compound of formula I, or a pharmaceutically acceptable salt thereof, in an amount effective to treat the diseaseStimulating luminal epithelial cells in the lung or middle/inner ear, eye, genitourinary, sperm cells; or beating frequency of cilia in certain cells in the immune system, including nucleophilic cells and macrophages: formula I

Wherein:

X₁、X₂and X₃Each independently is O^-Or S^-. Preferred X₂And X₃Is O^-。

R₁Is O, imido, methylene or dihalomethylene (e.g., dichloromethylene, difluoromethylene). Preferred R₁Is oxygen or difluoromethylene.

R₂Is H or Br. Preferred R₂Is H. Particularly preferred compounds of formula I are uridine 5 '-triphosphate (UTP) and uridine 5' -O- (3-thiotriphosphate) (UTP. gamma.S).

Formula I is a preferred embodiment of the compounds of the invention, but the method of the invention may also comprise administration of a compound of formula II (adenosine 5' -triphosphate [ ATP ]]Or 1, N⁶-ethano-adenosine triphosphate or adenosine 1-oxide triphosphate) or a compound of formula III (5' cytidine triphosphate [ CTP ]]Or 3, N⁴-ethanocytidine triphosphate), or a compound of formula iv (P)¹,P⁴-di (cytidine-5') tetraphosphate (A)₂P₄) Or P¹,P⁴Di (uridine-5') tetraphosphate (U)₂P₄). Formula II

Wherein:

R₁、X₁、X₂and X₃Are as defined in formula I.

When R is₂In the absence of a double bond (adenine) between N-1 and C-6, R₃And R₄Is H, or

When R is₂When there is a double bond (adenine 1-oxide) between O, N-1 and C-6, R₃And R₄Is H, or

R are linked together₃、R₄And R₂is-CH = CH-, N-6 and N-1 form a ring with the double bond between N-6 and C-6 (1, N)⁶-ethanoadenine). Formula III

Wherein:

R₁、X₁、X₂and X₃Are as defined in formula I.

When R is₇In the absence of a double bond (cytosine) between N-3 and C-4, R₅And R₆Is H, or

R are linked together₅、R₆And R₇is-CH = CH-, and the double bonds between N-3 and N-4 and N-3 and C-4 form a ring (3, N)⁴-ethanocytosine).

Formula IV

Wherein:

b is adenine or uracil.

A second aspect of the invention is a pharmaceutical formulation comprising a compound of formula I, II, III or IV in a pharmaceutically acceptable carrier in an amount effective to stimulate lung or middle and inner ear epithelial cells; the eye or urogenital tract; a sperm cell; an ovary or an oviduct; or ciliary beat frequency in certain cells of the immune system, including nucleophilic cells and macrophages.

In a third aspect of the invention, the active compounds disclosed herein are used in the preparation of a medicament for administration to a patient in need thereof for stimulation of epithelial cells in the lungs or middle and inner ear; the eye or urogenital tract; a sperm cell; an ovary or an oviduct; or ciliary beat frequency in certain cells of the immune system, including nucleophilic cells and macrophages.

Detailed description of specific embodiments

The present invention may be used to increase mucociliary clearance of secretions remaining in the lungs, sinuses or middle and inner ear in subjects in need of such treatment for any reason, including but not limited to. The methods of the invention may also be used to treat primary ciliary dyskinesia, secondary ciliary dyskinesia, catagmatic syndrome, otitis media, cystic fibrosis, disorders of the ciliary clearance barrier system of the eye or genitourinary mucociliary clearance resulting from impairment of ciliary movement, disorders of the immune system resulting from impairment of ciliary movement of nucleophilic cells and macrophages, hydrocephalus resulting from impairment of ciliary movement, male infertility resulting from impaired propulsion of sperm cilia, female infertility resulting from impaired ciliary movement on the luminal epithelial cells of the ovary or fallopian tube, or any other disorder resulting from impaired ciliary movement. The present invention increases mucociliary clearance in three ways: (1) increasing the beating frequency of cilia on the luminal epithelial cell surface, (2) increasing the secretion of goblet cell mucins, and (3) increasing the water secreted into the peripheral cilial liquid layer by the luminal epithelial cell. Mucins secreted by goblet cells form a layer on top of cilia; capture foreign particles (including viruses and bacteria); the mucin layer is transported by the wave-like motion of cilia, which is carried out by the composition and hydration of a peripheral liquid layer of cilia surrounding the cilia. Although the main aspect of the invention is to increase ciliary beat frequency in patients with ciliary dyskinesia where cilia are permanently immobile regardless of treatment, the active compounds of the invention are able to clear residual mucosal secretions by increasing water secretion into the peripheral cilial liquid layer and increasing mucin secretion from goblet cells.

In addition, the active compounds of the present invention promote the removal of any inhaled foreign material from the trachea, due to their ability to increase lung mucociliary clearance in normal humans. This is in many cases-biological weapons (such as the chemical weapon ricin); inhaling smoke; industrial exposure to inhaled toxins (leading to a full range of diseases such as silicosis, anthracosis and so-called pneumoconiosis); and the development of allergic reactions to absorbable particles such as pollen.

Furthermore, the ability of the compounds of the present invention to increase pulmonary clearance is also of benefit in the diagnosis of pulmonary diseases-in particular, it improves the quality of radioisotope scanning by removing secretions from the lungs which would otherwise obscure the vision of the ventilated part of the lungs. In radioisotope scanning, the mismatch of ventilation and perfusing the lung is used to identify areas of lung infarction. As a result, lung ventilation is improved upon administration of the compounds of the invention, and the scanning of the ventilation fraction will be clearer and more favorable for the diagnosis of true mismatches.

The invention is primarily directed to the treatment of human subjects, but may also be used in veterinary medicine for the treatment of other mammalian subjects, such as dogs and cats.

The compounds of formula I above include: (a) uridine 5' -triphosphate (UTP); (b) uridine 5' -O- (3-thiotriphosphate) (UTP γ S); and (c) 5-bromo-uridine 5' -triphosphate (5-BrUTP). These compounds are known or may be prepared by known methods, or by other methods which will be apparent to those skilled in the art. See generally N.Cusack and S.Hournani, Annals N.Y.Acad.Sci.603,172-81 (biological action entitled "extracellular ATP"). For example, UTP can be prepared by the method described by Kenner et al in journal of the chemical society (j.chem.soc.)1954,2288; or Hall and Khorana in journal of the American chemical society 76,5056 (1954). See Merck Index, monograph No. 9795 (11 th edition 1989). UTP γ S can be prepared by the methods disclosed in r.s. goody and f.eckstein, journal of the american chemical society 93,6252 (1971).

For simplicity, the uridine triphosphates active compounds of formulae I-IV herein are shown as naturally occurring in the D configuration, but unless otherwise specified, the invention also encompasses the L configuration and mixtures of the D and L configurations. Naturally occurring compounds of the D configuration are preferred.

The compound of formula II shown above comprises (a) 5'Adenosine Triphosphate (ATP) and (b)1, N⁶-ethano adenosine triphosphate. The compound of the above formula III comprises (a) cytidine 5' -triphosphate and (b)3, N⁴-ethanocytidine triphosphate. These compounds may be prepared according to known methods or other methods apparent to those skilled in the art. For example, standard methods for phosphorylation of nucleosides, such as D.Hoard and D.Ott, journal of the American chemical society (J.Am.chem.Soc.)87,1785-1788 (1965); m.yoshikawa et al, tetrahedron (tetrahedron lett.)5065-68(1967) and idem, bull.chem.soc. (japan) 42,3505-05 (1969); moffatt and H.Khorana, J.Chem.Soc. 83,649-59 (1961); and B.Fischer et al, J.Med.chem., 36,3937-46(1993), which is incorporated herein by reference. The ethanolic derivatives of cytidine and adenosine are prepared by known methods, such as: kotchetkov et al tetrahedron (tetrahedron Lett.)1993 (1971); barrio et al, department of biochemical and biophysical research (biochem. biophysis. res. commun.)46,597 (1972); secreist et al, Biochemistry 11,3499 (1972); bierndt et al, Nucleic Acids research (Nucleic Acids Res.)5,789 (1978); k. Koyasuga-Mikado et al chem.pharm.Bull. (Tokyo) 28,932 (1980). The thiophosphorus groups having α, β, and γ can be derived by the following methods or suitable methods: ludwig and f.eckstein, journal of organic chemistry (j.org.chem.)54,631-35 (1989); eckstein and r.goody, Biochemistry (Biochemistry)15,1685 (1976); goody and f.eckstein, journal of the american chemical society 93,6252 (1971).

R₁Is CCl₂And CF₂The compounds of formula I, II or III are prepared in a similar manner to that described in G.Blackburn et al, J.chem.Soc.Perkin Trans.I,1119-25 (1984). R is prepared by a method similar to that described in T.Myers et al, J.Am.chem.Soc.85,3292-95(1963)₁Is CH₂Compounds of formulae I, II, III.

The compound of formula IV comprises P¹,P⁴-bis (adenosine-5') tetraphosphate (A)₂P₄) Or P¹,P⁴-bis (uridine-5') tetraphosphate (U)₂P₄). These compounds can be prepared according to the following known methods or methodsVariants were prepared: zamecnik et al, Proc.Natl.Acad.Sci.USA 89,838-42 (1981); and k.ng and l.e.orgel, Nucleic Acids research (Nucleic Acids Res.)15(8),3572-80 (1987).

UTP, ATP, CTP, A₂P₄、3,N⁴-ethanocytidine triphosphate, 1, N⁶-ethano adenosine triphosphate, adenosine 1-oxide triphosphate, ATP γ S, ATP β S, ATP α S, AMPPCH₂P、AMPPNHP、N⁴-ethanocytidine and 1, N⁶Ethanoadenosines are commercially available, e.g. from Sigma Chemical Company (usa) PO Box 14508, st.

The active compounds of the formulae I to IV can be administered as such or as their pharmaceutically acceptable salts, e.g. alkali metal salts such as sodium or potassium salts, alkaline earth metal salts such as manganese, magnesium and calcium salts or ammonium and tetraalkylammonium salts (NX)₄ ⁺Where X is C_1-4Alkyl) is administered. Pharmaceutically acceptable salts are those which retain the desired biological activity of the parent compound without adverse toxicological effects.

The active compounds disclosed herein may be administered to the lungs, sinuses, ears, eyes, sperm, ovaries or fallopian tubes or genitourinary tracts by any suitable means, but it is preferred to apply the active compound in the form of a spray to the respiratory tract so that the active compound enters the lungs and reaches the affected body position with impaired ciliary movement either directly or via systemic absorption and circulation. The active compound may be aerosolized in various forms, such as, but not limited to, a dry powder inhaler, a metered dose inhaler, or a liquid/liquid suspension. In dry powder delivery systems, the UTP may be formulated alone or in admixture with diluents or carriers such as sugars (lactose, sucrose, trehalose, mannitol) and the like, wherein the compound may be intimately admixed with the matrix by vitrification or simply with a pulmonary or tracheal acceptable carrier or other acceptable excipient. Dry powders may be obtained by methods well known in the art, such as spray drying, milling, lyophilization, supercritical fluid preparation, or by controlled crystallization or precipitation.

The dose of active compound that stimulates ciliary beat frequency will depend on the condition being treated and the condition of the subject to which it is administered, but will generally be an effective amount sufficient to achieve a concentration of about 10 in the lung, sinuses, ear, eye or genitourinary surface of the active compound^-7-10^-1Mole/liter, more preferably about 10^-6To about 10^-1Amount of moles per liter of concentration.

The daily dose to promote fluid drainage may be divided into single unit doses or multiple unit doses depending on the solubility of the particular formulation of active compound administered. Preferably, the daily dose does not exceed four times per day.

Another method of administering an active compound to the lungs, sinuses, ears, eyes, or genitourinary tract of a patient to promote fluid/exudate drainage includes oral dosage forms of the active compound, and administration by pouring a liquid suspension of the active compound into the mouth of the patient or by ingestion of the pill into the patient.

Another method for administering an effective amount of the active compound to the lungs, sinuses, eyes or middle and inner ears involves administering a liquid/liquid suspension containing the active compound (a nasal spray, which is a solution of inhalable particles to be inhaled into the respiratory tract of a subject, or liquid nasal drops, or liquid eye drops) and preparing a nasal spray or nasal drops or liquid pharmaceutical composition of the active compound in eye drops by mixing the active compound with a suitable excipient known in the art, such as sterile, pyrogen-free water or sterile saline.

Another method of administering an active compound to the middle ear or eye involves the topical administration of the active compound in a form such as a cream or gel to the outer ear or eye which penetrates the middle ear or pupil of a human patient through the tympanic membrane or cornea.

Another method for administering active compounds to the middle ear, urogenital tract or eye involves the administration of the active compound in the form of an injectable formulation, either directly from the external ear through the tympanic membrane into the middle ear, into the urogenital tract or into the eye. This also involves the use of a patch containing UTP directly on the tympanic membrane.

Another method of administering active compounds to the lungs, sinuses, middle and inner ear, eye or urogenital tract involves suppositories of the active compound such that a therapeutically effective amount of the compound is systemically absorbed to the lungs, sinuses, middle ear, eye, urogenital tract or the male or female reproductive system.

Another method of administering the active compound involves instilling a gel, cream or liquid suspension of the active compound into the surgical site, such that a therapeutically effective amount of the compound reaches the lungs, sinuses, middle ear, eyes or urogenital tract.

Another method of administering an active compound involves administering the compound to sperm cells by topical, injection, or immersion, such that a therapeutically effective amount of the compound is in contact with sperm cells having impaired ciliary movement.

Another method of administering active compounds involves administering the active compound via a transdermal patch, wherein the active compound is delivered to the site of infection by local absorption or systemic absorption and circulation.

UTP and compounds of formulae i-iv are also therapeutically effective when used in combination with other agents useful in the treatment of ciliary dyskinesia, including, but not limited to: (ii) an antibiotic; a vaccine; decongestants, mucolytics; non-steroidal anti-inflammatory agents; a steroid; an antiviral agent; and bronchodilators. UTP can also be used in conjunction with gene therapy in development. UTP can also be combined with a recently discovered protein defensin (defensin) for therapeutic use

The following examples illustrate the invention in more detail. These examples are given for illustration only and are not intended to be limiting.

Experiment of

Example 1

Beating frequency of external excitated cilia

Will 10^-4M UTP was used for tracheal epithelial cells isolated from normal subjects. In four minutes, the beating frequency (CBF) of cilia increased by 76 ± 17% compared to the base (from 9.3 ± 0.23 to 16.12 ± 0.92Hz, n =7, p < 0.0001). Will 10^-4Similar results were obtained with M UTP on tracheal epithelial cells isolated from patients with gallbladder fibrosis. The CBF increased by 56 + -17% compared to the substrate (from 11.25 + -0.56H 2 to 16.1 + -1.45 Hz).

Example 2

Treatment of primary ciliary dyskinesia

Uridine 5' -triphosphate (UTP) was administered to patients diagnosed with Primary Ciliary Dyskinesia (PCD) as confirmed by electron microscopy analysis of the ultrastructure of cilia in living tissue from the nose. The effect of UTP was determined by measuring clearance of inhaled radiolabeled particles from the lungs by radionuclide scanning techniques with a gamma camera. (ii) technetium 99 m-labeled iron oxide for inhalation per subject: (⁹⁹Tc-Fe₂O₃) An aerosol formulation. The subject inhaled aerosol for approximately 5 minutes, then sat in front of a gamma camera, followed by 20 minutes for the subject to inhale either saline control (0.12% saline) or 10^-2M UTP lasted about 20 minutes. After this inhalation, subjects were still sitting in front of the gamma camera for 30 minutes to measure clearance of radiolabeled iron oxide. Aerosolized UTP improved cough clearance with technetium 99m compared to saline vehicle alone, showing the effect of aerosolized UTP in treating primary ciliary dyskinesia.

In the same study, the amount of sputum produced by inhalation of UTP control placebo was measured. Subjects were allowed to inhale placebo or UTP for 20 minutes using the method described above. During the next 30 minutes, subjects were subjected to 60-90 controlled coughs and matched sputum samples were collected from 8 out of 12 patients. The total volume of sputum was measured.

Monitoring heart rate, ECG rhythm strip before, during and after inhalation of full doseLead ii, blood pressure, oxyhemoglobin saturation measured by a pulse oximeter. All patients were monitored for any adverse reactions during all study periods, and the scans ended after 30-min at 24 hours from the start of inhalation of the study drug. As a result: cough clearance (%/min)Time interval (minutes)

TABLE 1

Cough clearance (%/min), whole lung PCD

Excipient pair UTP

This was a randomized, double-blind, placebo-controlled, crossover study on 12 patients with PCD, aged over 10 years old. The data reviewed in table i above show a significant increase in clearance of UTP to cough at 60-minute (p < 0.05) and 120-minute (p < 0.05) compared to placebo (excipient = V); the improvement was also significant at the 30-minute time point. These data are particularly striking in that effects are observed only with a single dose of UTP. For Table II, these data suggest a strong correlation between cough clearance and expectoration. Production of sputum (ml/min)

TABLE II sputum volume produced by PCD patients per unit time

Table II summarizes the increased volume of sputum produced per unit time by PCD patients by inhaling UTP. The study was double blind and involved 8 patients. Column 1 shows that about 0.2 ml of sputum is expelled per minute after saline inhalation. In contrast, column 2 showed that inhaled UTP produced a doubling of the amount of sputum excreted over the same time period (p < 0.01). UTP facilitates expectoration compared to vehicle based on questionnaires completed from the patient (done in blind rolls). Some patients state that sputum is thinner and more prone to expectoration following administration of UTP than following administration of vehicle. These data are particularly striking given that the only observed effect is after the present dose of UTP is administered. From the data in fig. 2, these data indicate a large correlation between cough clearance and expectoration amount.

Other technical embodiments

Although the invention has been shown and described with respect to illustrative embodiments, other embodiments will be apparent to those skilled in the art, and it should be understood that changes, omissions and additions may be made to the foregoing without departing from the spirit of the invention and the scope of the claims.

Claims (23)

1. A method of treating ciliary dyskinesia in a subject in need thereof, said method comprising:

administering to a subject a compound of formula I, II, III or IV, or a pharmaceutically acceptable salt thereof, in a pharmaceutical carrier in an amount effective to increase ciliary beat frequency in an affected area of the body: formula IWherein: x₁、X₂And X₃Each independently selected from OH or SH; r₁Selected from O, imido, methyleneAlkyl and dihalomethylene R₂Selected from H and Br; formula IIWherein: r₁、X₁、X₂And X₃Are as defined in formula I. When R is₂In the absence of a double bond (adenine) between N-1 and C-6, R₃And R₄Is H, or when R₂When there is a double bond (adenine 1-oxide) between O, N-1 and C-6, R₃And R₄Is H, or

R are linked together₃、R₄And R₂is-CH = CH-, N-6 and N-1 form a ring with the double bond between N-6 and C-6 (1, N)⁶-ethanoadenine). Formula III

Wherein:

R₁、X₁、X₂and X₃Are as defined in formula I.

When R is₇In the absence of a double bond (cytosine) between N-3 and C-4, R₅And R₆Is H, or

R are linked together₅、R₆And R₇is-CH = CH-, and the double bonds between N-3 and N-4 and N-3 and C-4 form a ring (3, N)⁴-ethanocytosine). Formula IV

Wherein: b is adenine or uracil.

2. The method of claim 1, wherein the compound is administered to a patient in need of such treatment and is delivered to the eye to treat a disorder of the mucociliary clearance system of the eye resulting from impaired ciliary movement.

3. The method of claim 1 wherein said compound is administered to a patient in need of such treatment for delivery to the urogenital tract for treatment of a mucociliary clearance system disorder of the urogenital tract resulting from impaired ciliary movement.

4. The method of claim 1 wherein said compound is administered to a patient in need of such treatment, released into the blood to treat immune disorders caused by impaired ciliary movement of nucleophilic cells and macrophages.

5. The method of claim 1, wherein said compound is administered to a patient in need of such treatment and is released from sperm to treat male infertility caused by impaired sperm ciliary movement.

6. The method of claim 1 wherein said compound is administered to a patient in need of such treatment and is delivered to the ovary or fallopian tube to treat infertility in women due to impaired ciliary movement of ovarian epithelial cells.

7. The method of claim 1, wherein said compound is administered to said subject by delivering to the naso-pharyngeal airways of said subject a liquid/liquid suspension of said compound, including nasal drops or sprays, such that a therapeutically effective amount of said compound contacts the lungs, sinuses, middle or inner ear, eyes, urogenital tract, sperm, ovaries and fallopian tubes, or damaged ciliary movement sites in nucleophilic cells and macrophages of said subject either directly or through systemic absorption and circulation.

8. The method of claim 1, wherein said compound is administered to said subject by delivering an oral form of said compound such that a therapeutically effective amount of said compound contacts damaged ciliary movement sites in the lungs, sinuses, middle or inner ear, eye, genitourinary tract, sperm, ovaries and fallopian tubes, or nucleophilic cells and macrophages of said subject either directly or through systemic absorption and circulation.

9. The method of claim 1, wherein said compound is administered to said subject by delivering an aerosol suspension of said compound such that a therapeutically effective amount of said compound contacts the damaged ciliary transport sites in the lungs, sinuses, middle or inner ear, eyes, genitourinary tract, sperm, ovaries and fallopian tubes, or nucleophilic cells and macrophages of said subject either directly or through systemic absorption and circulation.

10. The method of claim 1, wherein said compound is administered by delivery of a topical form of said compound to the middle ear or eye of said subject via the tympanic membrane or cornea such that a therapeutically effective amount of said compound contacts the locus of impaired ciliary movement in the luminal epithelial system or pupil of the middle or inner ear of said subject.

11. The method of claim 1, wherein said compound is administered to said subject by delivery of an injectable form of said compound such that a therapeutically effective amount of said compound contacts damaged ciliary motility sites in the lungs, sinuses, middle or inner ear, eyes, genitourinary tract, sperm, ovaries, and fallopian tubes, or nucleophilic cells and macrophages of said subject either directly or through systemic absorption and circulation.

12. The method of claim 1, wherein said compound is administered to said subject by delivery of a suppository form of said compound such that a therapeutically effective amount of said compound contacts the damaged ciliary transport sites in the lungs, sinuses, middle or inner ear, eyes, genitourinary tract, sperm, ovaries and fallopian tubes, or nucleophilic cells and macrophages of said subject either directly or through systemic absorption and circulation.

13. The method of claim 1, wherein said compound is administered to said subject by applying intra-operatively a release of said compound in the form of a gel, cream or liquid suspension, such that a therapeutically effective amount of said compound is in direct contact or through systemic absorption and circulation with the damaged ciliary movement sites in the lungs, sinuses, middle or inner ear, eye, genitourinary tract, sperm, ovary and fallopian tubes, or nucleophilic cells and macrophages of said subject.

14. The method of claim 1, wherein the compound is administered to the impregnate in vivo by local injection or impregnation of the compound such that a therapeutically effective amount of the compound is contacted with ciliated motility-impaired sperm.

15. The method of claim 1, wherein said compound is administered by transdermal patch delivery such that a therapeutically effective amount of said compound is in direct contact or in contact by local absorption and circulation with the impaired ciliary transport region of the ear, eye, lung or genitourinary tract of said subject.

16. The method of claim 1, wherein the compound is present at a concentration sufficient to achieve a concentration of about 10 on the surface of the lungs, sinuses, middle or inner ear, eye, genitourinary tract, sperm, ovary and fallopian tube, or nucleophilic cells and macrophages of the subject^-7To about 10^-1The amount of moles/liter.

17. The method of claim 1, wherein X₂And X₃Is OH.

18. The method of claim 1, wherein R₁Is oxygen.

19. The method of claim 1, wherein R₂Is H.

20. The method of claim 1, wherein the compound of formula i is selected from the group consisting of uridine 5 ' -triphosphate, uridine 5 ' -O- (3-thiotriphosphate), 5-bromo-uridine 5 ' -triphosphate and pharmaceutically acceptable salts thereof.

21. The method of claim 1, wherein the compound of formula ii is selected from the group consisting of adenosine 5' -triphosphate, 1, N⁴-ethano adenosine triphosphate, adenosine 1-oxide triphosphate and pharmaceutically acceptable salts thereof.

22. The method of claim 1, wherein the compound of formula iii is selected from cytidine 5' -triphosphate (CTP), 3, N⁴-ethanocytidine triphosphate and pharmaceutically acceptable salts thereof.

23. The method of claim 1, wherein the compound of formula iv is selected from P¹,P⁴-di (cytidine-5') tetraphosphate (A)₂P₄) And P¹,P⁴Di (uridine-5') tetraphosphate (U)₂P₄) And pharmaceutically acceptable salts thereof.