KR100549904B1 - A pharmaceutical composition for treating ciliary dyskinesia, containing uridinde triphosphate and related compounds - Google Patents

Abstract

The present invention relates to a method of stimulating the frequency of ciliary pulsations in a subject (patient) in need of treatment. The method of the present invention comprises an amount of trphophosphate nucleotide (eg, uridine 5′-triphosphate (UTP), an analog of UTP, or another analog thereof) sufficient to stimulate the ciliary pulsation frequency. Administering to the airways, eyes, ears or genitourinary system. This method can be used for ciliary dyskinesia, Cartagener's syndrom or other diseases (e.g., male infertility due to impairment of sperm anterior rush, or immune deficiency due to impaired cilia of neutrophils or macrophages). Useful for treating.

The present invention also relates to pharmaceutical preparations (pharmaceutical compositions) and methods for their preparation for stimulating the frequency of ciliary pulsations in a subject (patient). Methods of administering pharmaceutical formulations of the present invention include administration of liquid suspensions (including nasal sprays, nasal drops or eye drops), oral administration, inhalational administration by spray, topical administration, injection, suppository administration, intraoperative red wine (Iv) or in vitro direct application to sperm.

Description

Pharmaceutical composition for treating ciliary dyskinesia containing uridine triphosphate and related compounds {A PHARMACEUTICAL COMPOSITION FOR TREATING CILIARY DYSKINESIA, CONTAINING URIDINDE TRIPHOSPHATE AND RELATED COMPOUNDS}

The present invention stimulates a patient's ciliary beat frequency by administering a particular uridine, adenosine, or cytidine triphosphate, thereby reducing the mucous cilia of the secreted mucus secretion from the patient's lungs, sinuses or ears. Or to promote cough clearance. The present invention also relates to a medicinal composition or pharmaceutical preparation for treating ciliary dyskinesia which contains a specific uridine, adenosine or cytidine triphosphate as an active ingredient.

Mucociliary clearance (MCC) is an important defense mechanism of human airway and middle ear / inner ear. The harmonious pulsations of the cilia in the nose, trachea, bronchus and middle ear propel the mucus layer in the pharyngeal direction, carrying the microorganisms and other particles captured in the mucus. The normal function of such a system depends on the frequency and coordination of ciliary pulsations, and on the nature of the mucus. There are three components in the mucociliary cleaning system: (1) mucus layer formed by the secretion of mucin by goblet cells, and (2) the ciliary layer that transports the mucus layer deposited by synchronizing pulsation. And (3) a periciliary liquid layer that is less viscous than the mucin layer, surrounding the cilia and allowing free movement of the cilia. The concentration of electrolyte and water in the peri-cyliform liquid layer is controlled by luminal epithelial cells (R. Boucher et al., Adenosine and Adenine Nucleotides: From Molecular Biology to Integrative Physiology, p. 525-32 entitled "Mechanisms and Therapeutic Actions of Uridine Triphosphates in the Lung "(L. Belardinelli et al. Alumwer Academic Publishers, Boston 1995).

Primary ciliary dyskinesia (PCD) is a congenital disease characterized by ultrastructural defects and dyskinesia of the cilia, resulting in no or abnormal ciliary movement. The most common clinical manifestations of PCD are chronic respiratory diseases (eg, sinusitis, rhinitis and bronchiectasis) and otitis media. Because PCD patients have no or severely damaged mucociliary clearance (MCC), the only useful mechanism for cleaning (cleaning) or moving secretions is coughing. Cough clearance (cleaning rate) can be measured in a similar manner as described above for MCC. PCD also damages the anterior rush of sperm leading to male infertility (D. Schidlow, Ann Alergy 73 (b), 457-68 (1995)). PCD also impairs cell motility of certain immune system cells, including neutrophils and macrophage (N. Valerius, Eur J Clin Invest 13, 489-94 (1983)). PCD may be the cause of hydrocephalus caused by ciliary dysfunction (M. Greenstone, Arch Dis Child 59, 481-82 (1984)). The incidence of PCD was calculated to be 1 per 16,000 live births, with about 50% of infected individuals also having inversions (right heart). Bronchiectasis, sinusitis and inversion (right heart) are consistently referred to as Cartagener's syndrome (M. Sleigh, Lancet ii, 476 (1981)). It is hypothesized that Cartagener's syndrome is caused by a lack of ciliary movement in the fetus, which randomly rotates the intestine so that one half has an inversion (right heart) and the other has a normal cardia situs. (B. Afzelius Science 193, 317-19 (1976)). The clinical course of PCD is mainly characterized by early life-long sinus and ear infections that progressively change to adult lung / respiratory disease. Chronic respiratory tract infections can lead to chronic disturbance changes in lung tissue, progressive loss of lung function, and eventually death.

Secondary and more common forms of ciliary dyskinesia are acquired forms of the disease. Chronic inflammation caused by severe viral or bacterial respiratory infections, chronic smoking, severe air pollution, chemical or thermal lacerations to the airways, intubation and mechanical ventilation, near drowning, rupture of cell membranes, loss or incorporation of microtubules And ciliary structure, including the formation of compound cilia, which may result in abnormal or absent ciliary function (J. Ballenger Ann Otol Rhinol Laryngol 97 (3Pt. 1), 253-58 ( 1988); M. Pedersen Lung 168 Suppl., 368-76 (1990)). Respiratory infections that often cause secondary ciliary dyskinesia include ventilator-related pneumonia (VAP) in influenza, adult respiratory distress syndrome, and ICU patients. In some cases, acquired ciliary dyskinesia may be reversed by appropriate timely intervention, but permanent damage to these factors and / or long-term exposure renders the ciliary damage irreversible. Clinical manifestations and pathways have been shown to be similar to PCD with respect to chronic lung infections, progressive loss of lung function, and disruptive lung disease.

Typical mammalian respiratory epithelial cells contain about 200 cilia. Each cilia have 9 pairs of peripheral microtubules and two central tubes. Each pair of peripheral microtubules comprises a subunit A and a subunit B, each subunit having a set of bent arms attached to so-called inner dynein arms and outer dynein arms. Have These dynein arms contain ATPase, an enzyme that destroys adenosine triphosphate (ATP), which provides energy for cilia movement. The most common ultrastructural abnormality associated with primary ciliary dyskinesia is the absence of all dynein arms (B. Afzelius et al., J. Cell Biol 66, 225-32 (1975)). Application has begun to investigate whether ATPase can activate immobilized cilia in vitro (J. Forrest et al., Am Rev Resp Dis 120, 511-15 (1979)). Although the results were positive, they were not consistently replicated by others. It was later found that application of Ca ²⁺ and cAMP extracellularly could increase the pulsation frequency of respiratory cilia (A. Lansley et al., Am J. Physiol 263, L232-42 (1992)). It is not clearly defined that specific therapies will stimulate ciliary pulsations when complete ciliary immobility is demonstrated. In this case, it is therapeutically advantageous to increase the hydration of the viscous mucus secretions.

Applicants have found that extracellular triphosphate nucleotides, particularly uridine 5'-triphosphate (UTP), modulate all three components of the mucociliary transport system. UTP stimulates ciliary pulsation frequency (R. Boucher et al., Supra); UTP stimulates mucin secretion by embryonic cells (M. Lethem et al., Am J Respir Cell Mol Biol 9, 315-22 (1993)); UTP stimulates Cl ⁻ secretion in airway epithelial cells to increase hydration of the peri-cyliform liquid layer (M. Knowles et al., N Eng J. Med 325, 533-38 (1991)). In addition, Applicants have demonstrated that UTP is safe for PCD patients and improves cough cleaning (summary submitted to P. Noone et al., US Thoracic Society 1996 International Conference).

Various clinical manifestations of ciliary dyskinesia, such as absent or damaged mucociliary clearance in the respiratory tract and middle ear / inner ear, impaired anterior rush of sperm, and impaired motility of neutrophils and macrophages, are found in UTP and related compounds. Adenosine 5'-triphosphate (ATP); Cytidine 5'-triphosphate (CTP); 1, N ₆ -ethenoadenosine triphosphate; Adenosine 1-oxide triphosphate; 3, N ⁴ -ethenocytidine triphosphate; P ₁ , P ₄ -di (adenosine-5 ') tetraphosphate (A ₂ P ₄ ); Or other nucleoside phosphates, such as P ₁ , P ₄ -di (uridine-5 ′) tetraphosphate (U ₂ P ₄ ), may be ameliorated or alleviated by administering to the site of infection of the body.

Summary of the Invention

The present invention provides a method of treating ciliary dyskinesia in a subject (patient) in need of treatment. The method comprises administering to a patient an amount of a compound of Formula 1 or a pharmaceutically acceptable salt thereof effective to stimulate the frequency of ciliary pulsations (administrable sites include luminal epithelial cells of the lung or middle ear / inner ear, eye, urogenital; sperm cells; Or specific cells of the immune system, including neutrophils and macrophages.

[Formula 1]

(In Formula 1, X _1, X ₂ and X ₃ are each O ^- A, X ₂ and X ₃ are O ^{- -} or S, and is preferably,

R ₁ is O, imido, methylene, or dihalomethylene (eg, dichloromethylene, difluoromethylene), preferably R ₁ is oxygen or difluoromethylene,

It is preferred that R ₂ is H or Br and R ₂ is H, and particularly preferred compounds of formula 1 are uridine 5'-triphosphate (UTP) and uridine 5'-O- (3-thiotriphosphate) ( UTPγS)).

Formula 1 is a preferred embodiment of the compound, but the method of the present invention also provides a compound of Formula 2 (adenosine 5 'triphosphate [ATP] or 2, N ⁶ -ethenoadenosine triphosphate or adenosine 1-oxide triphosphate) or Compound of formula 3 (cytidine 5 'triphosphate [CTP] or 3, N ⁴ -ethenocytidine triphosphate) or compound of formula 4 (P ¹ , P ⁴ -di (adenosine-5') tetraphosphate (A ₂ P ₄ ) or P ¹ , P ⁴ di (uridine-5 ′) tetraphosphate (U ₂ P ₄ )).

[Formula 2]

(In Formula 2, R ₁ , X ₁ , X ₂ and X ₃ are as defined in Formula 1,

R ₃ and R ₄ are H while R ₂ is absent and there is a double bond between N-1 and C-6 (adenine), or

R ₃ and R ₄ are H while R ₂ is O and there is a double bond between N-1 and C-6 (adenine 1-oxide), or

R ₃ , R ₄ and R ₂ together form —CH═CH to form a ring from N-6 to N-1 with a double bond between N-6 and C-6 (1, N ⁶ -etheno Adenine)

[Formula 3]

(In Formula 3, R ₁ , X ₁ , X ₂ and X ₃ are as defined in Formula 1,

R ₅ and R ₆ are H, while R ₇ is absent and there is a double bond between N-3 and C-4 (cytosine), or

R ₅ , R ₆ and R ₇ together form —CH═CH to form a ring from N-3 to N-4 with a double bond between N-4 and C-4 (3, N ⁴ − Tenositocin))

[Formula 4]

(In Formula 4, B is adenine or uracil)

A second aspect of the present invention relates to luminal epithelial cells of the lung or middle ear / inner ear; Mucous clearance defense system of the eye or genitourinary system; Sperm cells; Ovary or fallopian tubes; Or a pharmaceutical formulation (pharmaceutical composition) comprising a compound of formula 1, 2, 3 or 4 and a pharmaceutically acceptable carrier or excipient in an amount effective to stimulate the frequency of ciliary pulsations in certain cells of the immune system, including neutrophils and macrophages.

A third aspect of the invention relates to luminal epithelial cells of the lung or middle ear / inner ear of a patient in need of treatment; Mucous clearance defense system of the eye or genitourinary system; Sperm cells; Ovary or fallopian tubes; Or the use of active compounds for the preparation of a medicament for therapeutically stimulating the frequency of ciliary pulsations in certain cells of the immune system, including neutrophils and macrophages.

The method of the present invention can be used to stimulate the frequency of ciliary pulsations in a subject (patient) in need of treatment, including but not limited to increasing mucosal clearance of the lungs, sinuses or suspended secretions in the middle and inner ear. Can be. The methods of the present invention also include dysfunction of the mucociliary clearance defense system of the eye or urogenital tract caused by primary ciliary dyskinesia, secondary ciliary dyskinesia, Cartagener syndrome, otitis media, cystic fibrosis, ciliary motor impairment. Disease, diseases of the immune system caused by impaired ciliary movement of neutrophils and macrophages, hydrocephalus caused by impaired ciliary movement, male infertility caused by impaired ciliary anterior rush of sperm, luminal epithelium of the ovary or fallopian tube It can be used to treat female infertility caused by impaired ciliary movement of cells or other diseases caused by impaired ciliary movement. The present invention increases mucociliary clearance in three ways: (1) to increase the frequency of ciliary pulsations of cilia on the surface of luminal epithelial cells, (2) to increase mucin secretion by germ cells, And (3) increasing the secretion of water into the peri-cyst liquid layer by luminal epithelial cells. Mucins secreted by germ cells form a layer on top of the cilia and capture foreign particles, including viruses and bacteria; The mucin layer is transported by the cilia's undulating action, and the cilia's movement is promoted by the composition (composition) and hydration of the liquid layer around the cilia. Although there is a major aspect of the invention in increasing the frequency of ciliary pulsation in patients who are unable to permanently exercise or suffer from ciliary dyskinesia, regardless of treatment, the active compounds of the present invention are characterized by the secretion of water into the peri And by increasing the secretion of mucin by embryonic cells, it is possible to promote the clearance of suspended mucus secretion.

In addition, because of the well-proven ability of the active compounds of the present invention to increase pulmonary mucus ciliary clearance in normal subjects (patients), the active compounds of the present invention accelerate the cleaning (purification) of certain types of foreign substances inhaled from the airways. You can. This includes a number of context-biological engagements, such as drug ricin for chemical engagement; Smoking inhalation; Industrial exposure to inhaled toxins (eg causing silicosis, carbonosis and other so-called pneumoconiosis in general); And allergic reactions to inhaled particles (eg pollen).

In addition, the ability of the active compounds of the present invention to increase lung clearance has proven to be beneficial in diagnosing lung disease, in particular in improving the quality of radioisotope injections in the lungs by eliminating secretions of obscure visualization of the lung vents. In radioisotope injection of the lungs, inadequate binding of the vented lungs to the perfused lungs is used to identify the site of pulmonary infarction. As a result of improved aeration of the lungs after administration of the active compounds of the invention, the aerated regions of the injections were further distinguished so that the diagnostics were in a better position to clearly identify the inadequate binding.

The present invention relates primarily to the treatment of human subjects, but may also be used for veterinary treatment in other mammals such as dogs and cats.

Examples of the compound of Formula 1 include: (a) uridine 5′-triphosphate (UTP); (b) uridine 5′-O- (3-thiotriphosphate) (UTPγS); And (c) 5-bromo-uridine 5'-triphosphate (5-BrUTP). The compounds may be known or prepared according to known methods and their variants will also be known in the art. Generally N. Cusack and S. Hourani, Annals N.Y. Acad. Sci. 603, 172-81 (named "biological activity of extracellular ATP"). For example, UTP is described in Kenner et al. J. Chem. Soc. 1954, 2288; Or Hall and Khorana, J. Am. Chem. Soc. 76, 5056 (1954). See Merck Index, Monograph No. 9795 (11th Ed. 1989). UTPγS is described by R. S. Goody and F. Eckstein, J. Am. Chem. Soc. 93, 6252 (1971).

For the sake of simplicity, the above Chemical Formulas 1 to 4 represent the active compounds of D-coordination present in nature, but the present invention also provides compounds of L-coordination, and mixtures of compounds of D-coordination and L-coordination compounds (herein Unless otherwise indicated, refers to a mixture of compound L-coordination compounds in the D-configuration. Preference is given to active compounds of D-coordination present in nature.

Examples of the compound of Formula 2 include (a) adenosine 5'-triphosphate (ATP) and (b) 1, N ⁶ -ethenoadenosine triphosphate. Compound examples of the compound of Formula 3 include (a) cytidine 5'-triphosphate and (b) 3, N ⁴ -ethenocytidine triphosphate. The compounds may be prepared according to known methods, variations of which will be apparent to those skilled in the art. For example, D. Hoard and D. Ott, J. Am. Chem. Soc. 87, 1785-1788 (1965); M. Yoshikawa et al., Tetrahedron Lett. 5065-68 (1967) and idem., Bull. Chem. Soc. (Jpn) 42, 3505-08 (1969); J. Moffatt and H. Khorana, J. Am. Chem. Soc. 83, 649-59 (1961); And in B. Fischer et al., J. Med. Chem. 36. Phosphorylation of nucleosides by standard methods such as 3937-46 (1993) and references therein. Etheno derivatives of cytidine and adenosine are described in N. Kotchetkov et al., Tetrahedron Lett. 1993 (1971); J. Barrio et al., Biochem. Biophys. Res. Commun. 46, 597 (1972); J. Secrist et al., Biochemistry 11, 3499 (1972); J. Bierndt et al. Nucleic Acids Res. 5, 789 (1978); K. Koyasuga-Mikado et al., Chem. Pharm. Bull. (Tokyo) 28, 932 (1980). Derivatives having α, β, γ thiophosphate groups are described in J. Ludwig and F. Eckstein, J. Org. Chem. 54, 631-35 (1989); F. Eckstein and R. Goody, Biochemistry 15, 1685 (1976); R. Goody and F. Eckstein, J. Am. Chem. Soc. 93, 6252 (1971), or by applying it.

Compounds of Formula 1, 2 or 3 (R ₁ is CCl ₂ and CF ₂ ) are described in G. Blackburn et al., J. Chem. Soc. Perkin Trans. I, 1119-25 (1984). Compounds of Formulas 1, 2, 3 (R ₁ is CH ₂ ) are described in T. Myers et al. J. Am. Chem. Soc. 85, 3292-95 (1963).

Examples of compounds of formula 4 include (P ¹ , P ⁴ -di (adenosine-5 ') tetraphosphate (A ₂ P ₄ ) or P ¹ , P ⁴ -di (uridine-5') tetraphosphate (U ₂ P ₄ ) The compounds include P. Zamecnik et al., Proc. Natl. Acad. Sci. USA 89, 838-42 (1981); and K. Ng and LE Orgel, Nucleic acid Res. 15 (8), 3572 It may be prepared according to the known method described by -80 (1987), or according to a variant thereof.

And UTP, ATP, CTP, A ₂ P ₄ , 3, N ⁴ -ethenocytidine triphosphate, 1, N ⁶ -ethenoadenine triphosphate, adenosine 1-oxide triphosphate, ATPγS, ATPβS, ATPαS, AMPPCH ₂ P, AMPPNHP, N ⁴ -ethenocytidine and 1, N ⁶ -ethenoadenosine are usually useful, for example, US Missouri ZIP Codes; 63178 Lewis is a product of Mailbox 14508 Sigma Chemical Company.

The active compounds of the formulas 1 to 4 are themselves or in the form of pharmaceutically acceptable salts thereof, for example alkali metal salts such as sodium or potassium, alkaline earth metal salts such as manganese, magnesium and calcium or ammonium and tetraalkyl ammonium salts, NX ₄ ⁺ In which X is a C _1-4 alkyl group. Pharmaceutically acceptable salts are salts which preserve the desired biological activity of the parent compound and do not confer undesired toxin effects.

The active compounds disclosed herein may be administered by various suitable means to the lungs, sinuses, ears, eyes, sperm, ovaries or fallopian tubes, or the genitourinary system, but are preferably administered in the respiratory tract in the form of a nebulized active compound This allows the active compound to enter the lungs and reach parts of the body that suffer from impaired ciliary movement, either directly or through systemic absorption and circulation. The active compound may be sprayed in various forms such as, but not limited to, dry powder inhalants, metered doses of inhalants or liquid / liquid suspensions. In the delivery (delivery) of dry powders, UTP can be combined alone or in combination with a diluent or carrier such as sugars (e.g., lactose, sucrose, trihalose, mannitol), and the active compounds can be glassified. Directly into the matrix or simply mixed with a carrier or other acceptable excipients for pulmonary or airway delivery. Dry powders can be obtained by methods known in the art, such as spray drying, milling, lyophilization, supercritical fluid preparation or controlled crystallization or precipitation.

The dosage of active compound to stimulate the frequency of ciliary pulsations will vary depending on the conditions of treatment and the subject's condition, but generally the effective amount is about 10 ^-7 at the surface of the patient's lungs, sinuses, ears, eyes, or genitourinary tract. To an amount sufficient to achieve a concentration of the active compound of from about 10 ⁻¹ mol / l, more preferably from about 10 ⁻⁶ to about 10 ⁻¹ mol / l.

Depending on the solubility of the particular agent of the active compound being administered, the daily dose that promotes flow release may be divided into one or several unit doses. The daily dose is preferably no more than four times a day.

Other methods of administering the active compound to the patient's lungs, sinuses, ears, eyes or genitourinary system to facilitate fluid / secretory discharge include administering the active compound in an oral dosage form, injecting the active compound into the patient's mouth. Administration to the patient in the form of a liquid suspension or tablets or pills swallowed by the patient.

Other methods of administering an effective amount of the active compound to the lungs, sinuses, eyes or middle ear and inner ear include liquid / liquid suspending agents (nasal sprays or eye drops of respirable particles or breathable combinations of the patient inhaling the active compound). )). Liquid pharmaceutical compositions of nasal sprays, or active compounds for the preparation of nasal drops or eye drops, can be prepared by combining with appropriate excipients (eg, sterile water or sterile salts without pyrogens) by techniques known to those skilled in the art. .

Another method of administering the active compound to the middle ear or eye involves administering a topical dosage form of the active compound administered as a cream or gel to the outer ear or eye, which continues to penetrate the patient's middle ear or lens through the eardrum or cornea. do.

Other methods of administering the active compound to the middle ear, genitourinary or eye include injecting the active compound, ie, directly into the middle ear from the outer ear, through the tympanic membrane, into the urogenital genus, or into the eye. This includes patches containing UTP that are applied directly to the tympanic membrane.

Other methods of administering the active compound to the lungs, sinuses, middle ear and inner ear, eyes or urogenitals include active compounds in which a therapeutically effective amount of the compound reaches through systemic absorption into the lungs, sinuses, middle ear, eyes, urogenital, male or female reproductive system. Administration of suppositories.

Other methods of administering the active compound include intraoperative red wine in the form of gels, creams or liquid suspensions of the active compound such that a therapeutically effective amount reaches the lung, sinus, middle and inner ear, eye or urogenital tract.

Another method of administering the active compound comprises ex vivo administration of the active compound to the sperm in the form of a topical dosage form, injection or acupuncture of the compound such that a therapeutically effective amount of the compound contacts the sperm with impaired cilia.

Another method of administering the active compound involves the administration of the active compound via a subcutaneous patch form in which the active compound is delivered to the affected area via local absorption or systemic absorption and circulation.

UTP and the compounds of formulas 1-4 also include other agents used to treat ciliary dyskinesia (e.g., antibiotics, vaccines, decongestants, mucolytic agents, nonsteroidal serous agents, steroid agents, anti-irritants and bronchodilators). Therapeutically advantageous when used in combination with, but not limited to, those inclusive. UTP can also be used in conjunction with other therapies in development (eg, gene therapy). UTP can also be used in conjunction with the recently discovered therapeutic protein, defensin.

The invention is explained in more detail in the following examples. The following examples are only intended to illustrate the present invention, and should not be construed as limiting the scope of the present invention by the following examples.

Example 1

In Vitro Stimulation of Cilia Pulsation Frequency

To the airway epithelial cells isolated from normal a subject (patient) was added to the UTP 10 ^-4 M. Within 4 minutes, the ciliary pulsation frequency (CBF) increased 76 ± 17% compared to baseline (9.3 ± 0.23 to 16.12 ± 0.92Hz, n = 7, p <0.0001). Similar results were obtained when 10 ⁻⁴ M of UTP was added to airway epithelial cells isolated from patients infected with cystic fibrosis. CBF increased 56 ± 17% compared to baseline (11.25 ± 0.56 to 16.1 ± 1.45 Hz).

Example 2

Treatment of Primary Cilia

Uridine 5'-triphosphate (UTP) was administered to a patient diagnosed with primary ciliary dyskinesia (PCD) (identified by electron microscopic analysis of the cilia's microscopic defects from the nasal biopsy). The efficacy of UTP was verified by measuring the cleanup rate (cleaning rate) of radio-labeled particles inhaled from the lungs by a radionuclide injection technique using a gamma camera. Each patient inhaled an aerosol of iron oxide labeled with technetium 99m ( ⁹⁹ Tc-Fe ₂ O ₃ ). Patients inhaled the aerosol for about 5 minutes. The patients then sat in front of the gamma camera and randomly inhaled a salt control (0.12% saline) or 10 ⁻² M UTP for about 20 minutes. After the inhalation, patients remained and sat in front of the gamma camera for the next 30 minutes to measure the cleanup rate (cleaning rate) of the radio-labeled iron oxide. The efficacy of nebulized UTP in treating primary ciliary dyskinesia was demonstrated by an improvement in cough clearance (cleaning rate) of technitium 99m compared to saline excipient alone.

In the same study, the amount of sputum induced by inhalation of UTP for placebo was measured. Patients inhaled placebo or UTP for 20 minutes according to the method above. Thereafter, the patients had a controlled cough 60 to 90 times in 30 minutes, and suitable sputum samples were collected from 8 of 12 patients. The total volume of the sputum was measured.

Safety data was collected by monitoring pre-inhalation, inhalation, post-inhalation heart rate, ECT rhythm strip-lead II, blood pressure, and oxygen hemoglobin saturation by pulse oximeter for all dosing periods. During all phases of the study all patients were examined for adverse reactions beginning with inhalation during drug study during each dosing period and ending after 30 minutes injection for 24 hours.

The results are shown in Figure 1, Table 1 shows the cough purification rate (cleaning rate) (% / min) [V is the case when the PCD excipient, U is administered UTP].

result:

This was a randomized, double-blind, placebo-controlled, crossover study in 12 PCD patients over 10 years of age. Table I above shows that UTP significantly improves cough clearance (cleaning rate) at 60-minute (p <0.05) and 120-minute (p <0.05) time points compared to placebo (excipient = V), thereby providing 30-minute time Data demonstrating significant improvement in this respect is summarized. This data is of particular note when the effects appear only after a single dose of UTP. With regard to the data in Table II, the data suggested a strong correlation between cough clearance (cleaning rate) and measures of sputum production.

Table 2 in FIG. 2 shows the volume of sputum produced per minute in PCD patients.

Table 2 summarizes the data demonstrating that the volume of sputum produced per unit time also increased in PCD patients by UTP inhalation. The study was blinded crossover study and included 8 patients. Column 1 shows that about 0.2 ml of sputum is produced per minute after inhalation of the salt excipient. In contrast, column 2 shows that the amount of sputum produced in the same time period (p <0.01) was about twice the amount of UTP inhalation. Based on data from a patient-complete questionnaire (completed in a blinded format), UTP has been shown to increase the ease of sputum generation compared to excipients. Some patients reported less sputum and eased after UTP inhalation than after excipient inhalation. These data are particularly noteworthy when the effect is seen only after administration of UTP alone. The data together with the data of FIG. 2 indicate that there is a strong correlation between the cough clearance (cleaning rate) and the measure of sputum production.

Another embodiment

While embodiments of the invention are illustrated in the above embodiments, other embodiments will be apparent to those skilled in the art. Accordingly, those skilled in the art should understand that the above embodiments can be variously changed, changed, and improved within the scope of the technical spirit described in the appended claims.

Claims (22)

A pharmaceutical composition for treating a primary ciliary dyskinesia of a subject (patient), comprising: (a) a compound of Formula I or Formula IV or a pharmaceutically acceptable salt thereof, and (b) a pharmaceutically acceptable carrier or Pharmaceutical compositions comprising excipients: (Formula 1) [In Formula I, X ₁ , X ₂ and X ₃ are each independently selected from the group consisting of OH and SH; R ₁ is selected from the group consisting of O (oxygen), imido, methylene, and dihalomethylene; R ₂ is selected from the group consisting of H and Br] (IV) [In Formula IV, B is uracil]. A pharmaceutical composition for treating dysfunction of a mucociliary clearance system of an eye of a subject, comprising: (a) a compound of formula (I) or formula (IV) or a pharmaceutical thereof; A pharmaceutical composition comprising an acceptable salt and (b) a pharmaceutically acceptable carrier or excipient: Formula I [In Formula I, X ₁ , X ₂ and X ₃ are each independently selected from the group consisting of OH and SH; R ₁ is selected from the group consisting of O (oxygen), imido, methylene, and dihalomethylene; R ₂ is selected from the group consisting of H and Br] (IV) [In Formula IV, B is uracil]. A pharmaceutical composition for treating dysfunction of a mucociliary cleaning system of a subject's urogenital tract, comprising: (a) a compound of formula (I) or formula (IV) or a pharmaceutically acceptable salt thereof; and (b) Pharmaceutical compositions comprising a pharmaceutically acceptable carrier or excipient: (Formula 1) [In Formula I, X ₁ , X ₂ and X ₃ are each independently selected from the group consisting of OH and SH; R ₁ is selected from the group consisting of O (oxygen), imido, methylene, and dihalomethylene; R ₂ is selected from the group consisting of H and Br] (IV) [In Formula IV, B is uracil]. A pharmaceutical composition for treating immune deficiency due to a disorder of ciliary movement of neutrophils and macrophages in a subject (patient) in need of treatment, comprising: (a) a compound of formula (I) or formula (IV) A pharmaceutical composition comprising a pharmaceutically acceptable salt and (b) a pharmaceutically acceptable carrier or excipient: (Formula 1) [In Formula I, X ₁ , X ₂ and X ₃ are each independently selected from the group consisting of OH and SH; R ₁ is selected from the group consisting of O (oxygen), imido, methylene, and dihalomethylene; R ₂ is selected from the group consisting of H and Br] (IV) [In Formula IV, B is uracil]. A pharmaceutical composition for treating male infertility due to a disorder of ciliary movement of sperm in a subject (patient) in need of treatment, comprising: (a) a compound of Formula I or Formula IV or a pharmaceutically acceptable salt thereof; and (b) a pharmaceutical composition comprising a pharmaceutically acceptable carrier or excipient: (Formula 1) [In Formula I, X ₁ , X ₂ and X ₃ are each independently selected from the group consisting of OH and SH; R ₁ is selected from the group consisting of O (oxygen), imido, methylene, and dihalomethylene; R ₂ is selected from the group consisting of H and Br] (Formula 4) [In Formula IV, B is uracil]. A pharmaceutical composition for treating female infertility due to a disorder of ciliary movement of ovarian or fallopian epithelial cells in a subject in need of treatment, comprising: (a) a compound of formula (I) or (IV) A pharmaceutical composition comprising a compound or a pharmaceutically acceptable salt thereof, and (b) a pharmaceutically acceptable carrier or excipient: (Formula 1) [In Formula I, X ₁ , X ₂ and X ₃ are each independently selected from the group consisting of OH and SH; R ₁ is selected from the group consisting of O (oxygen), imido, methylene, and dihalomethylene; R ₂ is selected from the group consisting of H and Br] (IV) [In Formula IV, B is uracil]. A pharmaceutical composition for stimulating sputum production in a subject, comprising: (a) a compound of formula (I) or formula (IV) or a pharmaceutically acceptable salt thereof, and (b) a pharmaceutically acceptable carrier or excipient: (Formula 1) [In Formula I, X ₁ , X ₂ and X ₃ are each independently selected from the group consisting of OH and SH; R ₁ is selected from the group consisting of O (oxygen), imido, methylene, and dihalomethylene; R ₂ is selected from the group consisting of H and Br] (IV) [In Formula IV, B is uracil]. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition is a formulation of a liquid / liquid suspending agent. The pharmaceutical composition according to any one of claims 1 to 7, wherein the pharmaceutical composition is a formulation of an oral dosage form. The pharmaceutical composition according to any one of claims 1 to 7, wherein the pharmaceutical composition is a formulation of an aerosol suspending agent. The pharmaceutical composition according to any one of claims 1 to 7, wherein the pharmaceutical composition is in the form of a topical formulation. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition is in the form of an injection. 8. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition is in the form of a suppository. 9. The pharmaceutical composition according to any one of claims 1 to 7, wherein the pharmaceutical composition is a formulation of a gel or cream. The pharmaceutical composition according to any one of claims 1 to 7, wherein the pharmaceutical composition is a formulation of aggression. The pharmaceutical composition according to any one of claims 1 to 7, wherein the pharmaceutical composition is a transdermal patch. 8. The pharmaceutical composition of claim 1, wherein X ₂ and X ₃ are OH. 9. 8. The pharmaceutical composition of claim 1, wherein R ₁ is oxygen. 9. 8. The pharmaceutical composition of claim 1, wherein R ₂ is H. 9. 8. The compound of claim 1, wherein the compound of Formula I is uridine 5′-triphosphate, uridine 5′-O- (3-thiotriphosphate) and 5-bromo-uri The pharmaceutical composition is selected from the group consisting of Dean 5'-triphosphate. 8. The pharmaceutical composition of claim 1, wherein the compound of formula IV is P ¹ , P ⁴ -di (uridine-5 ′) tetraphosphate (U ₂ P ₄ ). 9. The pharmaceutical composition according to any one of claims 1 to 7, wherein the amount of the compound of formula (I) or formula (IV) or a pharmaceutically acceptable salt thereof is 10 ^-4 M to 10 ^-1 M.