MXPA06003086A - Fused pentacyclic polyethers - Google Patents

Abstract

Disclosed are polycyclic polyether compounds of formula:(I) and pharmaceutical compositions comprising such compounds. Wherein R, OR1 and R2 are as defined herein. Also disclosed are methods of regulating mucus clearance in a cell, and methods of treating decreased mucus clearance or mucociliary dysfunction.

Description

PENTACICLIC FUSIONED POLYETHERS Field of the Invention The invention relates to polycyclic polyether compounds, pharmaceutical compositions comprising the compounds, and methods of treating diseases using the compounds and pharmaceutical compositions. More specifically, the invention relates to a naturally occurring, "brevenal" polycyclic polyether compound, isolated from the marine dinoflagellate K. brevis, and derivatives thereof. It also relates to pharmaceutical compositions and treatment methods for brevetoxin and ciguatoxin poisoning, and diseases characterized by decreasing mucus clearance and mucociliary dysfunction, which comprises the compounds and pharmaceutical compositions. Background of the Invention The decrease in mucus clearance is a pathological feature of diseases such as cystic fibrosis; chronic obstructive airway disease (also known as chronic obstructive pulmonary disease (COPD)) and asthma. The deteriorated clearance of mucus can also contribute to increase the incidence of pulmonary infections and obstruction of respiratory REF: 171511 pathways. In particular, cystic fibrosis is characterized by abnormal functioning of epithelial cells of the respiratory tract. Cystic fibrosis (or "CF") is caused by a defective gene encoding a Na + / Cl "transporter present on the surface of the epithelial cells that cover the airway passages of the lung and other organs. have identified hundreds of mutations, all of which result in deficient transport of sodium and chloride by epithelial cells.Cystic fibrosis (CF) is the most common autosomal recessive genetic disease in Caucasians that causes premature death in the United States. caused by mutations in chromosome 7, which encodes the cystic fibrosis transmembrane conductance regulator (CFTR) .The CFTR encodes an applicable membrane epithelial protein that functions both as a regulated cCAMP-chloride channel and a channel regulator. epithelial sodium Defects or absence of CFTR observed in CF patients can be seen as changes in the ultrastructure cilia, transport of the io n sodium and chlorine, and water transport through the epithelial cells of the respiratory tract. These changes can result in thick mucus and decreased mucociliary clearance leading to respiratory tract infections. This suggests that CFTR in normal lung tissue can regulate ENaC by low regulation of its conductance of sodium ions through the airway epithelium and decreased water transport in the cell resulting in less viscous mucus and mucociliary clearance unalterable The current treatment for CF has focused on several different therapies, but the most effective treatments to date are compounds that change the mucus viscosity and treat lung infections that arise when bacteria are trapped in the thick mucus. Therapies to increase mucociliary clearance generally direct two different strategies. The first is to regulate the absorption of sodium in apical epithelial cells using sodium channel blockers such as amiloride and its derivatives. Due to the decrease in sodium absorption, the transport of fluid in the epithelial cells is limited and the liquid volume of the surface normalizes. The second strategy is to use purinergic receptor binding compounds (ie, UTP and INS37217) which activate chloride secretion in airway epithelial cells, which in turn decreases sodium uptake and increases liquid surface volume . The purinergic receptors are also considered to regulate mucin secretion and they are involved in the activation of ciliary palpitation. By increasing the frequency of the ciliary beat, mucus transport could be increased which could clear bacteria and other particles from the lungs more quickly. These observations indicate that the activation of sodium channels can lead to defects in mucus clearance and bronchoconstriction, both of which are associated with respiratory diseases that include CF. If activation of voltage-gated sodium channels contributes to lung diseases, then modulation of voltage-gated sodium channels may be useful in alleviating airway pathologies associated with mucociliary dysfunction, such as asthma, chronic obstructive pulmonary diseases. , pulmonary infection (for example, pneumonia, pseudomonas), and cystic fibrosis. Thus, there is a need for active agents that can modulate the transport of water through the apical membranes of epithelial cells, these compounds can be useful in the regulation of mucus clearance, in addition as a treatment or prevention of conditions or diseases associated with mucociliary dysfunction. Florida's red tides are known to have adverse effects on both marine life and humans. These tides have been linked to large fish deaths, marine mammal mortality, and even human diseases. Human diseases caused by red tides include respiratory irritation through contact or inhalation and poisoning of neurotoxic shellfish (NSP) by consumption of exposed or contaminated marine food (Purkerson-Parker, et al., Chemistry and Biology, 2000 , 7: 385-393; Badén, D. G. et al., Toxicon, 1982; 20 (5): 929-932; Badén, D. G., and collaborators, Int. Rev. Cytol., 1983; 82: 99-150). Symptoms of NSP include nausea, vomiting, diarrhea, and choconstriction (Purkerson-Parker, et al., 2000). The provocative agent in the organisms of the red tide has been isolated and identified as brevetoxin. Fish poisoning by ciguatera (CFP, for its acronym in English) is a form of human poisoning caused by the consumption of subtropical and tropical marine fish that have accumulated toxins that occur naturally through their diet. Toxins are known to originate from several species of dinoflagellates (algae) that are common in ciguatera endemic regions at low latitudes. Marine fish most commonly implicated in fish poisoning by ciguatera include groupers, barracudas, red snapper, pompanos, mackerel, and triggerfish. Many other species of warm water fish can harbor ciguatera toxins. The occurrence of toxic fish is sporadic, and not all fish of a given species or given location will be toxic. Initial signs of poisoning occur within six hours after consumption of the toxic fish and commonly include a combination of gastrointestinal (eg, nausea, vomiting, and diarrhea), neurological disorders (eg, intensified paresthesia, arthralgia, myalgia, pain of head, inverse sensory temperature and acute sensitivity to extremes of temperature, vertigo, and muscle weakness), and cardiovascular disorders (for example, arrhythmia, bradycardia or tachycardia, and reduced blood pressure). The symptoms defined with these general categories vary with the geographical origin of the toxic fish. The diagnosis of PIC remains unsatisfied and is commonly based on the patient's symptoms and recent dietary history. Ciguatera poisoning is usually self-limiting, and the signs of poisoning often subside within several days of onset. However, in several cases the neurological symptoms are known to persist for weeks to months.In a few isolated cases the neurological symptoms have persisted for several years, and in other cases the recovered patients have experienced recurrence of neurological symptoms months or years later. of recovery, such relapses are often associated with changes in dietary habits or alcohol consumption, there is a low incidence of death resulting from respiratory and cardiovascular injury, and current treatments for fish poisoning by ciguatera are far from The intravenous administration of mannitol is commonly used but is usually only effective if it is used in the first 48-72 hours of exposure.Chronic CFP treatment is usually symptomatic.The CFP and NSP are thought to be induced by of binding at a common receptor site in known voltage-controlled sodium channels s as site 5. Linkage by brevetoxins or ciguatoxin at site 5 results in massive influx of sodium ions into normal resting potential. Thus, there is a need for active agents that can act as antagonists for linking brevetoxins or ciguatoxin to voltage-gated sodium channels, which are useful in the amelioration of neurological and gastrointestinal effects in people affected by NSP and CFP.

Brief Description of the Invention A fused pentacyclic polyether compound having activity as a brevetoxin antagonist has been discovered. This compound, Brevenal, is isolated from the purification of native sources, such as K. brevis, and other organisms of the red tide. Thus, in one aspect, the invention provides the Brevenal compound which can be represented by the following formula: In a broad aspect, the invention provides compounds of Formula I: and pharmaceutically acceptable salts thereof, wherein R is C1-C12 alkyl, C2-C12 alkenyl, C? -C? 2 alkyl esters, C? -C? 2 alkyl amides, C4-C? 2 alkenyl esters, alkylaryl esters C? -C? 2, C4-CX2 alkenylaryl esters, C4-C12 alkenyl amides, C? -C12 alkoxy, C? -C12 alkyl formyl, C2-C12 formyl alkenyl, C1-C12 alkanoylalkyl, alkanoyl C2-CX2 alkenyl, carboxy C alquilo-C12 alkyl, C2-C ?2 carboxy alkenyl, wherein the alkyl and alkenyl groups are optionally substituted with 1-6 substituent groups selected from the group consisting of: C3-C12 cycloalkyl, C3 heterocyclyl -C12, aryl, heteroaryl, C? -C6 alkyl, Cx-Cs alkoxy, halogen, C2-C6 alkenyl, OH, nucleosides, nucleotides, purines, pyrimidines, aromatic esters, aryl esters, cycloalkyl esters, cycloalkenyl esters, purines, or pyrimidines; ORi is OH or -0 (CO) CH3; and R2 is -CH = CHCH = CH2, -CH2-phenyl, or -CH2-pyridyl, wherein the phenyl and pyridyl groups are optionally substituted in each substitutable position with a group that is independently alkylCa-C6, alkoxyC-Ce , haloalkyl, haloalkoxy, hydroxy, hydroxyalkyl, halogen, -C02H, alkoxycarbonyl C? ~ -CG, -C (0) NH2, C (O) NH (C? -C6 alkyl), or -C (O) (C? -C6 alkyl) (C? -C6 alkyl) or pharmaceutically acceptable salts, solvates, esters, amides, hydrates, or combinations of the same. The compounds of the invention have activity as brevetoxin antagonists and are therefore useful in the treatment of brevetoxin and ciguatoxin poisoning. The present invention also provides compounds of Formula I wherein SZ S? H ot where Hal is chlorine, fluoro, iodine, or bromine; R3 is selected from H, OH, NH2, halogen, and N02, and Y is selected from CH, N, O, and S. The invention also relates to pharmaceutical compositions comprising a compound of the formula I, or a pharmaceutically salt acceptable, hydrate or solvate thereof, in combination with a pharmaceutically acceptable carrier, excipient, solvent, adjuvant or diluent. The compounds of the invention can also act as antagonists for the classes of compounds known as brevetoxins and ciguatoxin. The invention also relates to methods of regulating the rate of mucus clearance and treatment conditions or diseases associated with decreased clearance of mucus and mucociliary dysfunction in a subject, which comprises administering to a subject a compound of formula I or a pharmaceutically acceptable salt, solvate, or hydrate thereof. The invention also provides methods for the treatment of brevetoxin and ciguatoxin poisoning, which comprises administering to a subject a compound of the invention or a pharmaceutically acceptable salt, solvate, or hydrate thereof, in an amount effective to treat brevetoxin poisoning. or ciguatoxin. This treatment method of brevetoxin and ciguatoxin poisoning can help to prevent, treat, reduce the severity of, or delay the onset or progression of symptoms and disease states associated with brevetoxin and ciguatoxin poisoning.

Brief Description of the Figures Figure 1 illustrates the effect of brevenal concentrations (3, 10, and 100 pg / mL) on airway constriction induced by PbTx-3 in sheep. Figure 2 illustrates the effect of brevenal concentrations (3, 10, and 100 pg / mL) on airway constriction induced by PbTx-2 in sheep. Figure 3 illustrates the effect of brevenal (10 pg / mL) in airway constriction induced by hemi-brevetoxin in sheep. Figure 4 illustrates the effect of β-naphthoyl-PbTx-3 and brevenal (100 pg / mL) on reduction induced by PbTx-3 in tracheal mucus velocity (TMV) in sheep. Figure 5 illustrates the effect of PbTx-3 and PbTx-2 (CONC.10 pg / mL) on tracheal mucus velocity (TMV) in sheep.

Detailed Description of the Invention Definitions Unless defined otherwise, all scientific and technical terms used herein have the same meaning as will commonly be understood by one skilled in the art to which this invention pertains. All patents and publications referred to herein are incorporated by reference for all purposes. A "therapeutically effective" amount is defined as an amount effective to reduce or decrease at least one symptom of the disease being treated or to reduce or delay the onset of one or more clinical markers or symptoms of the disease. As used in this specification and the appended claims, the singular forms "a," "one," and "the" include plural referents unless the content clearly dictates otherwise. Thus, for example, it refers to a composition that contains "a compound" that includes a mixture of two or more compounds. It should also be noted that the term "or" is generally used in this sense including "and / or" unless the content clearly dictates otherwise. By "alkyl" and "C6-C6 alkyl" in the present invention means straight or branched chain alkyl groups having 1-6 carbon atoms, such as, methyl, ethyl, propyl, isopropyl, n-butyl, sec- butyl, tert-butyl, pentyl, 2-pentyl, isopentyl, neopentyl, hexyl, 2-hexyl, 3-hexyl, and 3-methylpentyl. It will be understood that in cases where an alkyl chain of a substituent (eg, of an alkyl, alkoxy or alkenyl group) is shorter or longer than 6 carbons, it will be so indicated in the second "C" as, for example, L - LO indicates a maximum of 10 carbons. By the term "halogen" in the present invention means fluorine, bromine, chlorine, and iodine. "Alkenyl" and "C2-Cn alkenyl" means straight and branched hydrocarbon groups having from 2 to n carbon atoms and from one to four double bonds and include, for example, ethenyl, propenyl, l-but-3-enyl , 1-pent-3-enyl, l-hex-5-enyl, (3E, 5E) -4,5-dimethyldeca-3, 5-diene and the like. As used herein, the term "cycloalkyl" refers to saturated carbocyclic groups having three to twelve carbon atoms. The cycloalkyl can be a monocyclic or polycyclic fused system. Examples of such groups include cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl. The cycloalkyl groups herein are unsubstituted or, as specified, substituted in one or more substitutable positions with several groups. For example, such cycloalkyl groups can be optionally substituted with, for example, Cx-C alkyl, C6-C6 alkoxy, halogen, hydroxy, cyano, nitro, amino, mono (C? -Cs) alkylamino, di (C? C6) alkylamino, C2-Ce alkenyl, C2-C6 alkynyl, C6-C6 haloalkyl, C6-6 haloalkoxy, amino (C6-C6) alkyl, mono (C6-C) alkylamino (C6-C6) alkyl or di (C? -Cs) alkylamino (C? -Ce) alkyl. By "aryl" is meant an aromatic carbocyclic group having a single ring (e.g., phenyl), multiple rings (e.g., biphenyl), or multiple condensation rings in which at least one is aromatic, (e.g., 1, 2, 3, 4-tetrahydronaphthyl, naphthyl), which is optionally mono-, di-, or trisubstituted. Preferred aryl groups of the present invention are phenyl, 1-naphthyl, 2-naphthyl, indanyl, indenyl, dihydronaphthyl, tetralinyl or 6,7,8,8-tetrahydro-5H-benzo [a] cycloheptenyl. The aryl groups herein are unsubstituted or, as specified, substituted in one or more substitutable positions with several groups. Preferred aryl groups are optionally substituted with Ci-Cg alkyl, C?-Cg alkoxy, halogen, hydroxy, cyano, nitro, amino, mono (C? -Ce) alkylamino, di (C-C3) alkylamino, C2-C6 alkenyl , C2-C6 alkynyl, C6-C haloalkyl, C6-C6 haloalkoxy, amino (C6Ce) alkyl, mono (C6C6) alkylamino (C6C6) alkyl or di (C6C6) alkylamino (C? -Cg) alkyl. Preferred aryl groups are phenyl and naphthyl, each of which is optionally substituted as described above. By "heteroaryl" is meant one or more 5-, 6-, or 7- ring aromatic ring systems which include fused ring systems of 9-11 atoms containing at least one and up to four heteroatoms selected from nitrogen , oxygen, or sulfur. Preferred heteroaryl groups of the present invention include pyridinyl, pyrimidinyl, quinolinyl, benzothienyl, indolyl, indolinyl, priidazinyl, pyrazinyl, isoindolyl, isoquinolyl, quinazolinyl, quinoxalinyl, phthalazinyl, imidazolyl, isoxazolyl, pyrazolyl, oxazolyl, thiazolyl, indolizinyl, indazolyl, benzothiazolyl. , beidazolyl, benzofuranyl, furanyl, thienyl, pyrrolyl, oxadiazolyl, thiadiazolyl, triazolyl, tetrazolyl, oxazolopyridinyl, imidazopyridinyl, isothiazolyl, naphthyridinyl, cinnolinyl, carbazolyl, beta-carbolinyl, isochromanyl, chromanyl, tetrahydroisoquinolinyl, isoindolinyl, isobenzotetrahydrofuranyl, isobenzotetrahydrothienyl, isobenzothienyl, benzoxazolyl , pyridopyridinyl, benzotetrahydrofuranyl, benzotetrahidrotienilo, purinyl, benzodioxolyl, triazinyl, phenoxazinyl, phenothiazinyl, pteridinyl, benzothiazolyl, imidazopyridinyl, imidazothiazolyl, dihidrobeoxazinilo, beoxazinilo, benzoxazinyl, dihidrobeotiazinilo , benzopyranyl, benzothiopyranyl, coumarinyl, isocoumarinyl, chromonyl, chromanonyl, pyridinyl-N-oxide, tetrahydroquinolinyl, dihydroquinolinyl, dihydroquinolinonyl, dihydroisoquinolinonyl, dihydrocoumarinyl, dihydroisocumarinyl, isoindolinonyl, benzodioxanil, benzoxazolinonyl, pyrrolyl N-oxide, pyrimidinyl N-oxide, pyridazinyl N- oxide, pyrazinyl N-oxide, quinolinyl N-oxide, indolyl N-oxide, indolinyl N-oxide, isoquinolyl N-oxide, quinazolinyl N-oxide, quinoxalinyl N-oxide, phthalazinyl N-oxide, imidazolyl N-oxide, isoxazolyl N- oxide, oxazolyl N-oxide, thiazolyl N-oxide, indolizinyl N-oxide, indazolyl N-oxide, benzothiazolyl N-oxide, beidazolyl N-oxide, pyrrolyl N-oxide, oxadiazolyl N-oxide, thiadiazolyl N-oxide, triazolyl N- oxide, tetrazolyl N-oxide, benzothiopyranyl S-oxide, benzothiopyranyl S, S-dioxide. More preferred heteroaryl groups include oxazolyl, isoxazolyl, pyridyl, pyrimidyl, pyridazinyl, and pyrazinyl. The heteroaryl groups herein are unsubstituted or, as specified, substituted in one or more substitutable positions with various groups. Preferred heteroaryl groups are optionally substituted with C? -Ce alkyl, C? -C3 alkoxy, halogen, hydroxy, cyano, nitro, amino, mono (C? -C6) alkylamino, di (C? -C6) alkylamino, C2 alkenyl -C6, C2-C6 alkynyl, C? -C6 haloalkyl, C? -C3 haloalkoxy, amino (C? C6) alkyl, mono (C? -C6) alkylamino (C? -C3) alkyl or di (C? Cg) alkylamino (C? -Ce) alkyl.

By "heterocycle", "heterocycloalkyl" or "heterocyclyl" means one or more 4-, 5-, 6-, or 7- ring carbocyclic ring systems which include fused ring systems of 9-11 atoms which they contain at least one and up to four heteroatoms selected from nitrogen, oxygen, or sulfur. Preferred heterocycles of the present invention include morpholinyl, thiomorpholinyl, thiomorpholinyl S-oxide, thiomorpholinyl S, S-dioxide, piperazinyl, homopiperazinyl, pyrrolidinyl, pyrrolinyl, tetrahydropyranyl, piperidinyl, tetrahydrofuranyl, pyrimidine-2, 4 (1H, 3H) -dione , 1H-benzo [d] imidazole-2 (3H) -one tetrahydrothienyl, homopiperidinyl, homomorpholinyl, homothiomorpholinyl, homothiomorpholinyl S, S-dioxide, oxazolidinonyl, dihydropyrazolyl, dihydropyrrolyl, dihydropyrazinyl, dihydropyridinyl, di-idropyrimidinyl, dihydrofuryl, dihydropyranyl, tetrahydrothienyl S- oxide, tetrahydrothienyl S, S-dioxide and homothiomorpholinyl S-oxide. The most preferred heterocyclic groups include lH-benzo [d] imidazole-2 (3H) -onyl, pyrimidine-2, 4 (1H, 3H) -dione, piperidinyl, pyrrolidinyl and piperazinyl. The heterocycle groups herein are unsubstituted or, as specified, substituted in one or more substitutable positions with various groups. Preferred heterocycle groups are optionally substituted with C? -C6 alkyl, C? -C6 alkoxy, halogen, hydroxy, cyano, nitro, amino, mono (C? -C6) alkylamino, di (C? -Cs) alkylamino, C2 alkenyl -C3, C2-C3 alkynyl, C? -C6 haloalkyl, C? -C6 haloalkoxy, amino (C? -C6) alkyl, mono (C? -C6) alkylamino (C? -C6) alkyl, di (C? C6) alkylamino (C? ~ Cs) alkyl or = 0. As used herein, the term "arylester" embraces aryloxycarbonyl and arylcarbonyloxy groups. As used herein, the term "alkyl ester" encompasses alkyloxycarbonyl and alkylcarbonyloxy groups. As used herein, alkylcarbonyl carries the same meaning as alkanoyl. As used herein, the term "alkylamide" embraces alkylaminocarbonyl groups, dialkylcarbonyl groups, and alkanoylamino groups. As used herein, the term "alkenyl amide" embraces alkenylaminocarbonyl groups, dialkenylcarbonyl groups, and alkenylcarbonylamino groups. As used herein, the term "alkenyl ester" embraces alkenyloxycarbonyl and alkenylcarbonyloxy groups. The term "alkylaryl ester" as used herein refers to alkyloxycarbonyl and acanoyloxy groups in which the alkyl portion carries an aryl or heteroaryl group.

The term "alkenylaryl ester" as used herein refers to alkenyloxycarbonyl and alkenylcarbonyloxy groups in which the alkenyl portion carries an aryl or heteroaryl group. The phrase "regulation of mucosal clearance" encompasses "controlling, promoting and / or influencing mucosal clearance." As used herein, the terms "treatment" and "treating" encompass the prophylactic administration of the compound or a composition Pharmaceutical comprising the compound ("prophylaxis") as well as therapy remedy for reducing or eliminating a disease or disorder mentioned herein. Prophylactic administration is intended to prevent disorders or prevention of the recurrence of disorders and can be used to treat a subject who is at risk of suffering from or having one or more disorders mentioned herein. Thus, as used herein, the term "treatment", or a derivative thereof, contemplates partial or complete inhibition of the established disease states, when an active ingredient of the invention is administered prophylactically or after initiation. of the disease state for which such active ingredient is administered. "Prophylaxis" refers to the administration of the active ingredient (s) to a mammal to protect the mammal from any of the disorders set forth herein, as well as others. As used herein, the term "subject" encompasses animals, including mammals and fish. Preferably the term refers to mammals such as humans, cattle and horses, more preferably to humans and domestic animals such as cats, dogs, and horses, and more preferably to humans. In one aspect, the present invention relates to compounds, or pharmaceutically acceptable salts thereof, of Formula (I): wherein R is C 1 -C 12 alkyl, C 2 -C 12 alkenyl, C 1 -C 12 alkyl esters, C 1 -C 12 alkyl amides, C 4 -C 12 alkenyl esters, Alkylaryl esters C 1 -C 4, C 4 -C alkenylaryl esters ? 2, C4-C? 2 alkenyl amides, C1-C12 alkoxy, C12 alkyl-formyl, C2-C2-C2-alkenyl formyl, C? -C12 alkanoyl, C2-C2-C2 alkanoyl, carboxyC? -C12 alkyl, carboxyC2-C? 2 alkenyl, wherein the alkyl and the alkenyl groups are optionally substituted with 1-6 substituent groups selected from the group consisting of: C3-C2 cycloalkyl, C3-Ci2 heterocyclyl, aryl, heteroaryl, C? -Cg alkyl, C? C3, halogen, C2-C6 alkenyl, OH, nucleosides, nucleotides, purines, pyrimidines, aromatic esters, aryl esters, cycloalkyl esters, cycloalkenyl esters, purines, or pyrimidines; 0R? is OH or -0 (C0) CH3; and R 2 is -CH = CHCH = CH 2, -CH 2 -phenyl, or -CH 2 -pyridyl, wherein the phenyl and pyridyl groups are optionally substituted with 1, 2, or 3 groups which are independently C 1 -C 3 alkyl, C 1 alkoxy -Cg, haloalkyl, haloalkoxy, hydroxy, hydroxyalkyl, halogen, -C02H, alkoxycarbonyl L-CS, C (O) NH2, -C (O) NH (C? -C6 alkyl), or -C (O) N (alkyl) Ci-Cg) (C? -C6 alkyl). In another broad aspect, Rx and R2 are as defined above and R is alkyl, halogen, alkenyl, cycloalkyl, aryl, heteroaryl, heterocycle, heterocyclealkyl or heterocyclyl. In still another aspect, R is C6-C12 alkyl, C6-C12 alkyl esters, C12-C12 alkyl amides, esters of C6-C2 alkylphenyl, C6-C6 alkoxy, C6-C6-cycloalkyl, C6-C alkanoyl. ? 2alkyl, carboxyC6-C? 2alkyl, wherein the alkyl portions are optionally substituted with 1-4 substituent groups selected from the group consisting of: C3-C6 cycloalkyl, C5-C6 heterocyclyl, pentyl, naphthyl, pyridyl, pyridyl, pyridazinyl, pyrazinyl, furanyl, thienyl, quinolinyl, indolyl, CX-C6 alkyl, Cx-C alkoxy, halogen, C2-Ce alkenyl, OH, nucleosides, nucleotides, purines, pyrimidines, phenyl esters, cycloalkyl esters, cycloalkenyl esters, purines, and pyrimidines, wherein each of the cyclic substituents on R is optionally further substituted with up to 5 groups which are independently C 1 -C 6 alkyl, C 1 -C 6 alkoxy, halogen, haloalkyl, haloalkoxy, hydroxy, C 1 -C 6 alkyl hydroxy, alkoxy C? -C4 C? -C6 alkyl, C02H, C (0) NH2, C (O) NH (Ca-C6 alkyl) , or C (0) N (Ci-Cg alkyl) (Cx-Cg alkyl). In still another aspect, R is C6-C2 alkenyl, C6-C2 alkenyl esters, C6-C2 alkenylphenyl esters, Ce-C2 alkenyl amides, C6-C12-alkenyl formyl, C6-C2 alkanoyl-alkyl. , alkanoylCe-C? alkenyl, carboxyC6-C12alkyl, carboxyCe-C? 2alkenyl, wherein the alkenyl groups are optionally substituted with 1-4 substituent groups selected from the group consisting of: C3-C6 cycloalkyl, C5-C6 heterocyclyl, phenyl, naphthyl, pyridyl, pyridyl, pyridazinyl, pyrazinyl, furanyl, thienyl, quinolinyl, indolyl, CC alkyl, Cx-Cg alkoxy, halogen, C2-Cs alkenyl, OH, nucleosides, nucleotides, purines, pyrimidines, phenyl esters, cycloalkyl esters esters of cycloalkenyl, purines, and pyrimidines, wherein each of the cyclic substituents on R are optionally further substituted with up to 5 groups which are independently C 1 -C 6 alkyl, C 1 -Cg alkoxy, halogen, haloalkyl, haloalkoxy, hydroxy , hydroxy Cx-Cg alkyl, alkoxy Ci-C4 alkyl C 1 -C 3 -C 0 2 H, C (0) NH 2, C (O) NH (C 1 -C 6 alkyl), or C (O) N (C 1 -C 6 alkyl) (C 1 -C 6 alkyl). In yet another aspect, OR? It is OH. In still yet another aspect, R2 is -CH = CHCH = CH2, -CH2 ~ phenyl, or -CH2-pyridyl, wherein the phenyl and pyridyl groups are optionally substituted with 1, 2, or 3 groups which are independently Cx- alkyl. Cg, C? -Cg alkoxy, haloalkyl, haloalkoxy, hydroxy, hydroxyalkyl, halogen, -C02H, C? -C6 alkoxycarbonyl, -C (0) NH2, -C (0) NH (C? -C6 alkyl), or - C (O) N (C? -Cg alkyl) (C? -C6 alkyl). In another aspect, R is C 1 -C 2 alkenyl substituted with 1, 2, or 3 groups that are independently C 1 -C 4 alkoxy, halogen, -CHO, -OCH 2 CH 20-, -OCH 2 CH 2 CH 20- or OH. In still another aspect, R is C 8 -C 2 alkenyl substituted with 1, 2, or 3 groups which are independently C 1 -C 4 alkoxy, halogen, -CHO, -0CH 2 CH 20-, -0CH 2 CH 2 CH 20- or OH. In still yet another aspect, R is C9-C10 alkenyl substituted with 1, 2, or 3 groups which are independently C1-C4 alkoxy, -CHO, -OCH2CH20-, -0CH2CH2CH20- or OH. In one aspect, when R is substituted with two alkoxy groups Ca-C4, these are the same. Still more preferably, when the two C?-C4 alkoxy groups are the same, they are bonded to the same carbon, thereby forming an acetal.

In yet another aspect, R is In yet another aspect, R is In yet another aspect, R is In another aspect, R is In another embodiment of this aspect, the compound is of Formula (I), wherein R is wherein Hal is chloro, fluoro, iodo, or bromine; R3 is selected from H, OH, NH2, halogen, and N02, and Y is selected from CH, N, O, and S. In another aspect, R2 is -CH = CHCH = CH2. In a preferred embodiment, R2 in the compound of Formula (I) is -CH = CHCH = CH2 in the cis conformation. In a preferred embodiment, R2 in the compound of Formula (I) is -CH = CHCH = CH2 in the cis conformation, and R is a C2-C12 alkenyl comprising a diene. In a more preferred embodiment, R2 in the compound of Formula (I) is -CH = CHCH = CH2 in the cis conformation, and R is a C2-C22 alkenyl comprising a diene, in the trans conformation. In another embodiment, R2 in the compound of Formula (I) is CH = CHCH = CH2 in the trans conformation, and R is a C8-C2 alkenyl comprising a diene, in the trans conformation. In another embodiment, R2 is -CH2-phenyl, or -CH2-pyridyl, wherein the phenyl and pyridyl groups are optionally substituted with 1, 2, or 3 groups which are independently C1-Cg alkyl, C6-C6 alkoxy, CF3, 0CF3, hydroxy, hydroxy C? -C6 alkyl, halogen, -C02H, alkoxycarbonyl C-Ce, -C (0) NH2, C (O) NH (C? -C6 alkyl), or -C (O) N (Cx-Cg alkyl) (Cz-C6 alkyl). In another embodiment, R2 is -CH = CHCH = CH2. In yet another mode, R2 is ^ J ^ Y "^ * - In yet another mode, R2 is In yet another modality, R is In another embodiment R and R2 are as defined above and Rx is H. In another preferred embodiment, the compound of Formula (I) is: In another preferred embodiment, the compound of the Formula In another preferred embodiment, the compound of Formula s: In another preferred embodiment, the compound of Formula s: In another preferred embodiment, the compound of Formula s: In another preferred embodiment, the compound of Formula s: In another preferred embodiment, the compound of Formula s: In another preferred embodiment, the compound of Formula s: In another preferred embodiment, the compound of Formula s: In another preferred embodiment, the compound of Formula s: In another preferred embodiment, the compound of Formula s: In another preferred embodiment, the compound of Formula s: In another preferred embodiment, the compound of Formula s: In another preferred embodiment, the compound of Formula s: In another preferred embodiment, the compound of the Formula In another preferred embodiment, the compound of Formula s: In another preferred embodiment, the compound of Formula s: In another preferred embodiment, the compound of Formula s: In one aspect of this preferred embodiment, the compound of Formula (I) is: In another preferred embodiment, the compound of Formula (I) is: In one aspect of this preferred embodiment, the compound of Formula (I) is: In another preferred embodiment, the compound of Formula (I) is: In one aspect of this preferred embodiment, the compound of Formula (I) is: In another preferred embodiment, the compound of Formula (I) is: The compounds of Formula (I) can have asymmetric centers and be presented as racemates, the racemic mixtures and as individual diastereomers, or enantiomers. All isomeric forms are included within the scope of the present invention. In a preferred aspect, the compounds of the invention are based on the core of the brevenal, that is, each stereocenter in the compounds of Formula (I) have the same configuration as the stereocenters in brevenal. In another aspect, the invention relates to pharmaceutical compositions comprising a compound of the formula I or a pharmaceutically acceptable salt, solvate, or hydrate thereof, and a pharmaceutically acceptable carrier, excipient, solvent, adjuvant or diluent. In another aspect, the invention provides methods for regulating mucus clearance, which comprise administering to a subject, or contacting a cell with, a compound, salt, solvate or hydrate of the invention, or a pharmaceutical composition comprising a compound, salt, hydrate, or solvate of the invention, in an amount effective to regulate the clearance of mucus in the subject or cell. As used herein, decreases in mucus clearance or mucociliary dysfunction are generally measured by tracheal mucus transport (TMV), a surrogate marker for complete lung clearance. In another aspect, the invention provides methods for the treatment of conditions or diseases related to, or associated with, decreased mucus clearance which comprises administering to a subject a compound of the invention, or pharmaceutical composition comprising a compound of the invention or a pharmaceutically acceptable salt, hydrate, ester, amide solvate, or mixtures thereof, in an amount effective to treat the condition or disease. This method of treating conditions or diseases associated with decreased mucus clearance can help prevent, treat, reduce the severity of, or delay the onset or progression of symptoms and disease states associated with decreased mucus clearance. Such conditions or diseases include non-limiting examples of chronic obstructive airway disease (also known as chronic obstructive pulmonary disease (COPD)); asthma; cystic fibrosis, bronchoconstriction, and other lung diseases; including lung diseases, such as the non-limiting examples pneumonia, Pseudomonas, and bronchitis; and cystic fibrosis. In one embodiment, the method of treatment can be used to treat chronic obstructive pulmonary diseases, such as emphysema, pulmonary fibrosis, and / or cough of smokers. In one embodiment, the treatment method can be used to treat asthma. In one embodiment, the method of treatment can be used to treat lung disease. In one embodiment, the method of treatment can be used to treat lung infection, including, but not limited to, pneumonia, or Pseudomonas. In one embodiment, the treatment method can be used to treat envetoxin by brevetoxins or ciguatoxin, and the symptoms thereof. In a preferred embodiment, the method of treatment can be used where the disease is cystic fibrosis. In another aspect, the invention provides methods for the treatment of symptoms related to conditions or diseases associated with decreased mucus clearance or mucociliary dysfunction, comprises administering to a subject in need of such treatment, a therapeutically effective amount of a compound of the invention, or a pharmaceutically acceptable salt, hydrate, or solvate thereof. In one embodiment of this aspect the methods of the invention can be used to treat diseases associated with mucous membranes, including such diseases involving the pancreas, intestines, kidneys, fallopian tubes, and / or vas deferens. In another embodiment of this aspect, the method may optionally comprise, in combination with a compound of the formula (I) or a pharmaceutically acceptable salt, solvate, or hydrate thereof, an effective amount of a compound known to be useful for the treatment of conditions or diseases associated with decreased clearance of mucus. The methods of the invention may optionally comprise additional therapeutic regimen such as adjuvant or supportive therapy. In one embodiment of the methods of the invention, the subject is an animal. More preferably, the animal is a mammal. Still more preferably, the mammal is a human. In another embodiment, the animal is a companion animal, such as, for example, a horse, dog or cat. The therapeutically effective amounts of the compounds of the invention, suitable for the methods are generally from about 0.1 pg / day to about 1000 mg / day. Therapeutically effective amounts will vary in accordance with various parameters including, for example, the particular therapeutic used and the physical characteristics of the subject / patient, and are in accordance with the knowledge of those skilled in the art. In a preferred aspect, the therapeutically effective amount for oral administration is from about 1 mg / day to about 1000 mg / day. In another aspect, from about 1 mg / day to about 500 mg / day. In yet another aspect, from about 1 mg / day to about 100 mg / day. In yet another aspect, from about 0.1 mg / day to about 10 mg / day. In another aspect, the therapeutically effective amount is administered in a dose of between about 1 mg to about 500 mg per dose. In yet another aspect, the therapeutically effective amount comprises a dose of between about 1 mg to about 100 mg per dose. Preferred dosages for administration by inhalation are from about 0.1 pg to about 1 pg per day. Preferred doses for administration by injection, that is, parenteral administration, are from about 100 ng to about 1 mg per day. The present invention also includes the use of a compound of the formula (I), or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for use in the treatment of a subject who has, or in the prevention of a subject from the development, NSP and / or CFP and symptoms associated with those poisonings, and who needs such treatment. In one aspect, this use of a compound of the formula (I) can be employed where the disease or condition is chronic obstructive pulmonary disease. In another aspect, this use of a compound of the formula (I) can be employed where the disease or condition is asthma.

In another aspect, this use of a compound of the formula (I) can be employed where the disease or condition is lung disease. In another aspect, this use of a compound of the formula (I) can be employed where the disease or condition is lung infection. In another aspect, this use of a compound of the formula (I) may be employed where the disease or condition is cystic fibrosis. In another aspect, this use of a compound of the formula (I) can be employed where the disease or condition is chronic bronchitis. In another aspect, this use of a compound of the formula (I) can be employed where the disease or condition is Karteneger's syndrome. In another aspect, this use of a compound of the formula (I) can be used where the disease or condition is bronchiectasis. In another aspect, this use of a compound of the formula (I) can be used where the disease or condition is an industry-related disease caused or aggravated by inhalation of gases, textile particles, grains of sand, or other industrial particles or fumes. Specific examples of particles and grains of sand include, for example, iron oxides, silica, talc, carbon, graphite, fibers, wood dust, grain dust, organic solvents and pollutant gases. In yet another aspect, the compounds of Formula (I) and pharmaceutical compositions comprising formula (I) may be employed where the disease or condition results from inhalation of bacteria or other pathogenic particles, for example, fungicidal particles. In this manner, the invention also encompasses methods of purifying pathogenic particles, such as particles comprising bacteria, eg, anthrax or fungal particles. The present invention also includes a container kit that includes a plurality of containers, each container includes one or more unit doses of a compound of the formula (I), or a pharmaceutically acceptable salt thereof. In one embodiment, this container kit includes each container adapted for oral administration and includes a tablet, gel, or capsule or inhaler. In one embodiment, this container kit includes each container adapted for parenteral administration and includes a product from the reservoir, syringe, ampule, or vial. In one embodiment, this container kit includes each container adapted for topical administration and includes a patch, medicated towels, ointment, or cream. The compounds of Formula (I) can form salts when they react with appropriate acids or bases. Pharmaceutically acceptable salts are generally preferred over the corresponding compounds of formula (I) since they often produce compounds that are usually more water soluble, stable and / or more crystalline. The pharmaceutically acceptable salts are any salt which retains the activity of the parent compound and does not impart any undesirable or detrimental effect to the subject to whom it is administered and in the context in which it is administered. The pharmaceutically acceptable salts include acid addition salts of both inorganic and organic acids. Preferred pharmaceutically acceptable salts include salts such as those described by Berge, Bighley, and Monkhouse, J. Pharm. Sci., 1977, 66, 1-19. Such salts can be formed from inorganic and organic acids. Representative examples thereof include maleic, fumaric, benzoic, ascorbic, pamoic, succinic, bismethylene-salicylic, methanesulfonic, ethanedisulfonic, acetic, propionic, tartaric, salicylic, citric, gluconic, aspartic, stearic, palmitic, itaconic, glycolic, p-aminobenzoic acids , glutamic, benzenesulfonic, hydrochloric, hydrobromic, sulfuric, cyclohexyl sulphonic, phosphoric and nitric. For other acceptable salts, see Int. J.

Pharm., 33, 201-217 (1986). The hydrates and solvates of the compounds together with the polymorphs thereof are also forms of the compounds of the invention and can be formed in accordance with techniques known to one of ordinary skill in the pharmaceutical art. The invention further includes complexes, particularly organometallic complexes, of the compounds of the invention. Complexes that can be prepared when processes known in the art are appropriately used.

Methods of the Invention Compounds of the invention, pharmaceutical formulations comprising the compounds, and pharmaceutically acceptable salts thereof, are useful for the treatment of a subject, preferably a mammal, more preferably a human, suffering from a disease or associated condition with decreased clearance of mucus, and are useful to help prevent or delay the onset of such disease or condition. The compounds and formulations of the invention are particularly useful for treating, preventing, or slowing the progression of chronic obstructive pulmonary disease, asthma, lung disease, lung infection, and cystic fibrosis. When treating and preventing a disease and condition associated with decreased clearance of mucus, and associated symptoms, the compounds of the invention may be used either individually or in combination, as is best for the subject. With respect to these diseases and conditions, the term "treat" means that the compounds of the invention can be used in subjects, preferably human subjects / patients, with existing condition or disease. The compounds of the invention will not necessarily cure the subject who has the disease but delays or retards the progression or further prevents the progression of the disease in such a way as to give the individual a more useful life. The term "prevents" means that if the compounds of the invention are administered to those who do not have the disease, or symptoms of the condition, but who would normally develop the disease or be at increased risk for the disease, the disease would not develop. In addition, "preventing" also includes delaying the development of the disease in an individual who ultimately develops the disease or is at risk for the disease due to age, family history, genetics or chromosomal abnormalities, and / or due to the presence of one or more biological markers for the disease. By retarding the onset of the disease, the compounds of the invention can prevent the individual from contracting the disease during the period in which the individual could normally contract the disease or reduce the rate of disease development or some of these effects but for the administration of compounds of the invention until the time in which the individual ultimately contracts the disease. Prevention also includes the administration of the compounds of the invention to those individuals who are predisposed to the disease. In a preferred aspect, the compounds of the invention are useful for slowing the progression of disease symptoms. In another preferred aspect, the compounds of the invention are useful for the prevention of further progression of the symptoms of the disease. In treating or preventing the above diseases, the compounds of the invention are administered in a therapeutically effective amount. The therapeutically effective amount may vary depending on the particular compound used, the physical characteristics of the subject to be treated, and the routes of administration, as is known to those skilled in the art. In the treatment of a subject retarding any of the conditions diagnosed above, a physician can administer a compound of the invention immediately and continue the administration indefinitely, as needed.

Forms and Amounts of Dosage The compounds of the invention can be administered by inhalation, orally, parenterally, (IV, IM, depo-IM, SQ, and depo SQ), sublingually, intranasally, intrathecally, topically, vaginally, or rectally. Dosage forms known to those of skill in the art are suitable for the delivery of the compounds of the invention. The compositions are provided so as to contain therapeutically effective amounts of the compounds of the invention. The compounds are preferably formulated into pharmaceutically suitable preparations such as tablets, capsules, or elixirs for oral administration or in sterile solutions or suspensions for parenteral administration. Commonly the compounds described above are formulated in pharmaceutical compositions using techniques and procedures well known in the art. About 1 pg to about 100 mg of a compound or mixture of compounds of the invention or a pharmaceutically acceptable salt or ester is compounded with a pharmaceutically acceptable carrier, carrier, excipient, binder, stabilizer, flavor, etc., in a dosage form of unit so called by accepted pharmaceutical practice. The amount of active substance in those compositions or preparations is such that an appropriate dosage is obtained in the indicated range. The compositions are preferably formulated in a unit dosage form, each dosage containing from about 0.1 pg to about 100 mg, preferably about 0.1 pg to about 10 mg, more preferably about 0.1 pg to about 10 pg, or about 1 pg to about 10 mg, of the active ingredient. The term "unit dosage form" refers to discrete physically appropriate units as unit doses for human subjects and other mammals, each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect, in association with a pharmaceutical excipient. appropriate. To prepare pharmaceutically acceptable compositions of the invention, one or more compounds of the invention are mixed with a suitable pharmaceutically acceptable carrier. In the mixture or addition of the compound or compounds, the resulting mixture can be a solution, suspension, emulsion or the like. Liposomal suspensions may also be appropriate as pharmaceutically acceptable carriers. These can be prepared according to methods known to those skilled in the art. The shape of the resulting mixture depends on a number of factors, including the projected mode of administration and the solubility of the compound in the carrier or vehicle selected. The effective concentration is sufficient to decrease or alleviate at least one symptom of the disease, disorder, or condition treated and can be determined empirically. Suitable carriers or pharmaceutical carriers for administration of the compounds provided herein include any such carrier known to those skilled in the art to be appropriate for the particular mode of administration. In addition, the active materials can also be mixed with other active materials that do not damage the desired action, or with materials that complement the desired action, or have another action. The compounds can be formulated as the pharmaceutically active ingredient alone in the composition or can be combined with other active ingredients. When the compounds exhibit insufficient solubility, the methods for solubilization can be used. Such methods are known and include, but are not limited to, use of cosolvents such as dimethylsulfoxide (DMSO), use of surfactants such as Tween, and dissolution in aqueous sodium bicarbonate. Derivatives of the compounds, such as salts or prodrugs, can also be used in the formulation of effective pharmaceutical compositions.

The concentration of the compound is effective for delivery of an amount on administration that decreases or alleviates at least one symptom of the disorder for which the compound is administered. Commonly, the compositions are formulated for single dosage administration. The compounds of the invention can be prepared with carriers that protect against rapid elimination from the body, such as time release formulations or coatings. Such carriers include controlled release formulations, such as, but not limited to, microencapsulated delivery systems. The active compound is included in the pharmaceutically acceptable carrier in an amount sufficient to exert a therapeutically useful effect in the absence of undesired side effects in the treated subject. The therapeutically effective concentration can be determined empirically by testing the compounds in known in vitro and in vivo model systems for the treated disorder. The compounds and compositions of the invention can be enclosed in multiple or single dose containers. The enclosed compounds and compositions can be provided in kits, for example, which include component parts that can be assembled for use. For example, an inhibitor compound in lyophilized form and an appropriate diluent may be provided as separate components for combination prior to use. A kit can include an inhibitor compound and a second therapeutic agent for co-administration. The inhibitor and the second therapeutic agent can be provided as separate component parts. A kit can include a plurality of containers, each container maintains one or more unit doses of the compound of the invention. The containers are preferably adapted for the desired mode of administration, including, but not limited to tablets, gel capsules, sustained release capsules, and the like for oral administration; reservoir products, prefilled syringes, ampoules, flasks, and the like for parenteral administration; and patches, medical pads, creams, and the like for topical administration. The concentration of the active compound in the composition of the drug will depend on the route of administration, and the distribution, metabolism, and rates of excretion of the compound, in addition to the dosage schedule, and the amount administered, together with other factors known to those skilled in the art. experience in the technique. The active ingredient can be administered once, or it can be divided into a number of smaller doses to be administered at time intervals. It is understood that the dosage and precise duration of the treatment is a function of the disease being treated and can be determined empirically using known evaluation protocols or by extrapolation of in vivo or in vitro test data. It should be noted that the concentrations and dosage values may also vary with the severity of the condition to be alleviated. Furthermore, it should be understood that for any particular subject, the specific dosage regimens must be adjusted in time according to the individual need and the professional adjustment of the administration of the person or the supervision of the administration of the compositions, and that the ranges of concentration set forth herein are exemplary only and are not intended to limit the scope or practice of the claimed compositions. If oral administration is desired, the compound should be provided in a composition that protects it from the acidic environment of the stomach. For example, the composition can be formulated in an enteric coating that maintains its integrity in the stomach and releases the active compound in the intestine. The composition can also be formulated in combination with an antacid and another ingredient. Oral compositions will generally include an inert diluent or an edible carrier and can be compressed into tablets or enclosed in gelatin capsules. For the purpose of oral therapeutic administration, the compound or the active compounds can be incorporated with excipients and used in the form of tablets, capsules, or tablets. The pharmaceutically compatible binding agents and adjuvant materials can be included as part of the composition. Tablets, pills, capsules, pills, and the like can contain any of the following ingredients or compounds of a similar nature: a binder such as, but not limited to, gum tragacanth, acacia, corn starch, or gelatin; an excipient such as microcrystalline cellulose, starch, or lactose; a disintegrating agent such as, but not limited to, alginic acid and corn starch, a lubricant such as, but not limited to, magnesium stearate; a flow improver agent, such as, but not limited to, colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; and a flavoring agent such as peppermint, methyl salicylate, or fruit flavor. When the dosage unit form is a capsule, it may contain, in addition to the material of the above type, a liquid carrier such as a fatty acid. In addition, the dosage unit forms may contain various other materials, which modify the physical form of the dosage unit, for example, coatings of sugar and other enteric agents. The compounds can also be administered as a component of an elixir, suspension, syrup, wafer, chewing gum or the like. A syrup may contain, in addition to the active compounds, sucrose as a sweetening agent and certain preservatives, dyes and dyes, and flavorings. The active materials can also be mixed or stirred with other pharmaceutically acceptable agents that do not damage the desired action, or with materials that complement the desired action. Solutions or suspensions used for parenteral, intradermal, subcutaneous, or topical application can include any of the following components: a sterile diluent such as water for injection, saline solution, stabilized oil, a naturally occurring vegetable oil such as sesame oil, coconut oil, peanut oil, cottonseed oil, and the like, or a synthetic fatty vehicle such as ethyl oleate, and the like, polyethylene glycol, glycerin, propylene glycol, or other synthetic solvent; antimicrobial agents such as benzyl alcohol and methyl parabens; antioxidants such as ascorbic acid and sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid (EDTA); buffer solutions such as acetates, citrates, and phosphates; and agents for tonicity adjustment such as sodium chloride and dextrose. Parenteral preparations may be enclosed in ampules, disposable syringes, or multiple dose vials made of glass, plastic, or other suitable material. Buffering, preservative, antioxidant, and the like can be incorporated as required. When administered intravenously, suitable carriers include physiological saline, phosphate buffered saline (PBS), and solutions containing thickening and solubilizing agents such as glucose, polyethylene glycol, polypropylene glycol, and mixtures thereof. Liposomal suspensions that include tissue-directed liposomes may also be suitable as pharmaceutically acceptable carriers. These can be prepared according to known methods for example, as described in US Patent No. 4,522, 811. The active compounds can be prepared with carriers that protect the compound against rapid elimination from the body, such as release formulations. the weather or coatings. Such carriers include controlled release formulations, such as, but not limited to, microencapsulated delivery systems and implants, and biocompatible and biodegradable polymers such as collagen, ethylene vinyl acetate, polyanhydrides, polyglycolic acid, polyorthoesters, polylactic acid, and the like. Methods for preparing the formulations are known to those skilled in the art.

The compounds of the invention can be administered by inhalation (either orally or intranasally), orally, parenterally (IV, IM, depo-IM, SQ, and depo-SQ), sublingually, intrathecally, topically, or rectally. Dosage forms for those skilled in the art are suitable for delivery of the compounds of the invention. The compounds of the invention can be administered enterally or parenterally. When administered orally, the compounds of the invention can be administered in usual dosage forms for oral administration as is well known to those skilled in the art. These dosage forms include the usual solid unit dosage forms of tablets and capsules in addition to liquid dosage forms such as solutions, suspensions, and elixirs. When the solid dosage forms are used, it is preferred that they be of the sustained release type such that the compounds of the invention need to be administered only once or twice a day. The oral dosage forms are administered to the subject 1, 2, 3, or 4, or more times per day or as necessary. It is preferred that the compounds of the invention are administered either three or less times, more preferably once or twice a day. Hence, it is preferred that the compounds of the invention are administered in oral dosage form. It is preferred that any oral dosage form be used, that it be designed to protect the compounds of the invention from the acidic environment of the stomach. Enteric coated tablets are well known to those skilled in the art. In addition, capsules filled with small spheres each coated to protect from stomach acid are also well known to those skilled in the art. As noted above, depending on whether the asymmetric carbon atoms are present, the compounds of the invention may be present as mixtures of isomers, as racemates, or in the form of pure isomers. The salts of the compounds are preferably pharmaceutically acceptable or non-toxic salts of the compounds of the formula I. For purposes of isolation and purification it is also possible to use pharmaceutically unacceptable salts.

Examples Example 1 Synthesis of the compounds Brevenal isolation The Brevenal can be isolated and purified from native sources, such as K. brevis, or other red tide organisms. Appropriate purification methodologies are well known in the art. See, for example, Badén et al, 1981, Toxicon 19: 455-463; Poli et al., Molecular Pharmacology, 1986 30: 129-135. The following procedure is representative. Brevenal is extracted from K. brevis cultures (Provasoli-Guillard National Center for Culture of Marine Phytoplankton, West Boothbay Harbor, ME) using chloroform. The chloroform layers are collected, dried and divided between petroleum ether and aqueous methanol to remove pigments and cellular lipid debris. The aqueous methanol layer (90%) is dried under vacuum and the components are separated using a column of silica gel (mobile phase CHC13: methanol: acetic acid; 100: 10: 1 w / w). Brevenal and brevetoxins are eluted together from the silica column; Fractions containing these materials are collected and combined. A low pressure column of matrix C18 is used to separate the remaining pigments from brevetoxins and brevenal using a mobile phase of acetonitrile: water (80:20 v / v), creating a "clarified" extract. The clarified extract is applied to a CLAR column, such as a reverse phase C18 Variant column (0.8 x 25 cm) with a run buffer of 90:10 methanol: water at a suitable pump ratio (eg 3- 4 mL / min). The detection of the elution peaks can be made by any method known in the art, such as UV detection at 215 nm. The peaks of interest are isolated and applied to another column, such as a hydrophobic interaction column (Hl) (e.g., Phenomenex C18 phenyl-hexyl column 0.8 x 25 cm) in suitable run buffer (e.g., 99% MeOH: 1% H20). The fractions containing the compounds of interest are pooled and the compounds isolated by any known method such as crystallization, solvent evaporation (Roto-Vap). The brevenal structure was determined using a number of spectroscopic methods including NMR, mass spectroscopy and FT-IR. The exact mass of brevenal as determined by high resolution mass spectroscopy is 656.4043. The primary and stereospecific structures were elucidated using 1-D and 2-D NMR spectroscopy in four different solvents. Various synthetic methodologies can be used to make compounds of the invention; Brevenal is a suitable starting material. Appropriate methodologies are known in the art. Brevenal can be used as a starting material as it is or can first be protected or converted to the corresponding alcohol or carboxylic acid for further elaboration. Representative synthetic procedures for preparing compounds of the invention from such starting materials are described in, for example, Mende, T.J., et al., Tetr. Lett., 1990; 31 (37): 5307-5310; Trainer, V.L. , et al., Molec. Pharm., 1991; 40 (6): 988-994; Keck, G.E., et al., Tetrahedron Lett., 1987, 28: 139-142; Alvarez, E., et al., Chem. Rev., 1995, 95: 1953-1980; Rein, et al., 1994: (a) J. Org Chem., 59: 2107-2113; (b) J. Org. Chem .. 59: 2101-2106. Each of these references is incorporated herein by reference in its entirety. Those skilled in the art will appreciate that minor modifications can be made to particular methods to arrive at the compounds of the invention.

Example 1 Derivative of -Amiloride Amiloride (N-amidino-3,5-diamino-6-chloropyrazinecarboxamide) exists in charge form under physiological conditions as shown below.

Structure of ßmiloride under Ssxta8a Tetrotíßioxlna physiological conditions Amiloride, Saxitoxin and Tetrodotoxin Structures The amiloride functionalization of brevenal: The introduction of a portion of guanidine in the brevenal side chain is carried out as follows. Brevenal is treated with sodium chlorite and sodium diacid phosphate in a tert-butanol / 2-methyl-2-butene solvent mixture to carry the acidic derivative 2 (reaction scheme 2). Treatment of 2 with CDI in DMF followed by addition of guanidine (free base) or guanidine carbonate provided the guanidine compound 3. ! } Í.B »0H fKí0 / R? I | N? TíjP0 ajl-mem-Mitio Scheme of reaction 2: Introduction of guanidine in the side chain of unsaturated aldehyde a, β of brevenal.

Preparation of benzamil guanidine brevenal: The synthesis is performed by following essentially the same procedure as described for the synthesis of 3 with the addition of benzamyl guanidine instead of guanidine (reaction scheme 2). Alternatively, Brevenal can be converted into the corresponding acid chloride to effect the conversion to the guanidine derivatives.

Example 2 Derivative of ß-naphthoyl ß-naftoil brevenal is prepared as follows. A solution of carbonyldiimidazole and 2-naphthoic acid in benzene is added to brevenol and refluxed overnight to give 3-naphthoyl brevenal 5 (reaction scheme 3).

S Derivative of ß-Naltoil Brevenal Reaction Scheme 3: Synthesis of β-naphthoyl derivatives The Brevenal is treated with sodium borohydride and cerium chloride in a mixture of DMF / methanol. After an ether extraction, the crude extract is purified in HPLC to give brevenol in good yield. Alternatively, the brevenal can first be converted to the corresponding carboxylic acid and subsequently reacted to the reaction with an appropriate alcohol to provide an ester.

Example 3 Aromatic brevenal A. The introduction of the hydroxy benzyl portion is carried out through a Grignard reaction between phenylmagnesium bromide and brevenal (reaction scheme 4). The extraction followed by purification provided an adduct of the ephedrine of brevenal 6.

Reaction Scheme 4: Synthesis of a brevenal ephedrine model B. A one-step synthesis can be used to prepare various benzene compounds. Using a Wittig reaction and commercially available phosphoranes, various derivatives comprising a phenyl group can be made. In this way, the treatment of phosphorans la-lt with base followed by addition of brevenal supplies the corresponding ethylenic compounds 8a-bf (reaction scheme 5). The compounds can also be purified by HPLC. c; R = m-amino; Y = H d; R = p-chloro; Y = H e: R = p-nitro; Y = H f: R = H; Y = N Reaction scheme 5: Synthesis of benzene derivatives EXAMPLE 4 Benzimidazolone Derivatives The synthesis begins by the activation of brevenol within a mesylate intermediate followed by the displacement of the mesylate by iodide to produce the iodo-brevenal derivative 9 (reaction scheme 6). Then the treatment of the commercially available 1, 3-dihydro-benzimidazl-2-one with sodium hydride in DMF followed by the addition of iodine-brevenal 9 supplying, after extraction and purification, the benzimidazolone-brevenal derivative 10.

B? Cvsnol 9 iodine derivative Benzoimldazolone Derivative Reaction Scheme 6: Synthesis of a benvenimidazolone model of brevenal Example 5 Brevenal nucleoside model ATP or UTP is treated with dicyclohexylcarbodiimide in DMF to generate the reactive portion which is then added to brevenol. The reaction mixture is heated under reflux and extracted. Purification of the crude extract provides the nucleotide derivatives 11 and 12 (reaction scheme 7).

Brevenol Scheme of reaction 7: Synthesis of UTP- and ATP-Brevenal Example 6 Experimental Protocols of Mechanisms in Respiratory Tracts Measurement of mechanisms in airways - Without sedating the sheep are locked in a car in the prone position with their heads immobilized. After topical anesthesia of the nasal passages with 2% lidocaine solution, a balloon catheter advances through a nostril into the lower esophagus. The animals are intubated with a covered endotracheal tube through the other nostril using a flexible fiber optic bronchoscope. The pleural pressure is estimated with an esophageal balloon catheter (filled with one ml of air) that is placed 5-10 cm from the gastroesophageal junction. In this position the pleural pressure of the expiratory end is in the range between -2 and -5 cm H20. Once the balloon is placed, it is secured so that it remains in position for the duration of the experiment. The lateral pressure in the trachea is measured with a lateral orifice catheter (internal dimension, 2.5 mm) advancing through and distal to the end of the endotracheal tube. The transpulmonary pressure, the difference between tracheal and pleural pressure, is measured with a differential pressure transducer catheter system which shows no phase shift between pressure and flow up to a frequency of 9 Hz. For pulmonary resistance (LR) measurement, the proximal end of the endotracheal tube is connected to a pneumotachograph (Fleisch, Dyna Sciences, Blue Bell, PA). Flow signals and transpulmonary pressure are recorded in an oscilloscope recorder which is linked to a computer for the online calculation of RL of transpulmonary pressure, respiratory volume (obtained by digital integration) and flow by iso technique. -volume. The analysis of 5-10 breaths is used for the determination of RL (Abraham et al., 1994).

Aerosol delivery systems - All aerosols are generated using an available medical nebulizer (Raindrop®, Puritan Bennett, Lenexa, KS) that provides an aerosol with an aerodynamic diameter of average mass of 3.2 μm (geometric SD 1.9) as determined by an Andersen cascade fixer.

The nebulizer is connected to a dosimeter system, which consists of a solenoid valve and a source of compressed air (20 psi = 1,406 kg / cm2). The outlet of the nebulizer was directed into a plastic part T, one end of which is connected to the inhaler port of a respirator Harvard The solenoid valve is activated for one second at the start of the inhaler respirator cycle. The aerosols were delivered in a tidal volume of 500 ml and a ratio of 20 breaths per minute (Abraham et al., 1994).

Respiratory sensitivity - To determine airway sensitivity, cumulative dose-response curves for carbacol were performed by measuring RL immediately after inhalation of buffer and after each consecutive administration of 10 breaths of concentrations in increase in carbacol (0.25, 0.5, 1.0, 2.0 and 4.0% w / v saline buffer). The challenge test is discontinued when RL is increased over 400% from the post-saline value or after the highest concentration of carbachol that has been administered. Respiratory sensitivity is estimated by determining the cumulative dose of carbachol in units of respiration (BU) that increases RL by 400% (PC400) by interpolation from the dose response curve. A unit of respiration (BU) is defined as 1 breath of an aerosol solution containing 1% w / v carbacol (Abraham et al., 1994).

Resistance of the nasal airways - Resistance of the nasal airway (NAR) is measured with a modified mask rhinomanometric technique. The head of the sheep is placed in a plexiglass cover with accessories for a front plate containing a pneumotachograph to measure the flow and two catheter ports to measure the pressure differential between the mouth and nose pressure (Abraham et al., 1998).

Tracheal mucus velocity - The sheep are intubated nasally with a 7.5 cm endotracheal tube, a diameter cut by 6 cm, after topical anesthesia of the nasal passages with 2% lidocaine solution. The tube cover is placed just below the buccal cords (verified by fluoroscopy) in order to allow maximum exposure of the tracheal surface area. TMV is measured in vivo by a roentgenographic technique. Between 10 and 20 disks of bismuth trioxide and radiopaque Teflon, diameter 1-mm, thickness 0.8-mm and 1.8 mg in weight, are insufflated in the trachea by means of the endotracheal tube. The axial speeds towards the head of the individual discs are recorded on videotape of a portable unit of the image enhancer. The individual disc speeds are calculated by measuring the distance traveled by each disc during a period of 1 minute of observation. For each run, the average value of all individual disk speeds is calculated. A collar containing radiopaque reference markers of known length are used by sheep, and are used as a standard to correct for magnification effects inherent in the fluoroscopy unit (O'Riordan et al., 1997).

Statistical Analysis - If the data is normally distributed, then the parametric statistics are used; if the data does not conform to a normal distribution, nonparametric statistics are used. Basic statistics tests include analysis of variance (ANOVA), that is, one-way ANOVA or two-way ANOVA with repeated measures for multi-point analysis, and unpaired or paired t test for adequate single point analysis . The nonparametric counterparts of these tests are: a) the Mann-Whitney test, which is the counterpart of the unpaired t-test; b) signed classification test of Wilcoxon, the counterpart of the paired test; c) Friedman Variance analysis for related samples, this is randomized block design; d) the Quade test, also a chance test of the block design but for use with small blocks (n <4); e) the Kruskal-Wallis test, ANOVA for unrelated samples; and f) a nonparametric pairwise comparison, analogous to the parametric Newman-Kuels pair test. When applied, the linear regression analysis is performed by the least squares method, and the correlations will be tested with the Spearman rho test. For all studies, an importance is accepted with p < 0.05 in an analysis of two branches (Conover, 1980).

Example 7 Fish bioassay Male mosquito fish (n = 104) were used in this test. The fish were placed individually in 50 mL beakers containing 20 L of water. The test compounds (PbTx-2 and brevenal) were dissolved in EtOH at a concentration of 0.1 mg / mL and added to the fish in a total of 200 μL EtOH. Control fish receive 200 μL EtOH. Fish were exposed to the toxin alone (1 μg / mL water), brevenal alone (1 or 2 μg / mL water), or both brevenal (1.0 μg / mL water and toxin (1.0 μg / mL water) with the brevenal being 3 minutes before the toxin was added, after the addition of the different compounds the fish were monitored for 24 hours or until the time of their death.The significant differences were determined using a two-way Student's t-test. PbTx-2 only died within about 7.5 minutes, while the fish exposed to control, or brevenal at 1 or 2 μg / mL did not die after 24 hours.The brevenal effectively protects the fish, from an equal concentration of PbTx-2, prolonging life by ~ 2.5-fold (with dead fish around 17 minutes after exposure) This suggests that brevenal is not toxic at concentrations effective for brevetoxin antagonist activity.

Example 8 Competitive rat brain synaptosome assays were performed as described in Poli et al., 1986. The data shown for titrated brevetoxin PbTx-2 were efficiently displaced (-80%) by brevenal and brief di-O-Me when the compounds were added at about 1000 x the concentration of PbTx-2 titrated. The approximate EC50 for each is: 3.53 nM (Ki = 1.76 nM) for PbTx-2; 3.69 uM (K¡. = 1.86 μM) per brevenal; 1.35 μM (Ki = 0.68 μM) for brevenal di-O-Me. The invention has been described with reference to various specific and preferred modalities and techniques. However, it will be understood that any of the variations and modifications may be made while they are within the spirit and scope of the invention. It is noted that in relation to this date, the best method known to the applicant to carry out the aforementioned invention, is that which is clear from the present description of the invention.

Claims (24)

Claims Having described the invention as above, the content of the following claims is claimed as property.

1. A compound of the Formula: characterized in that R is C? -C? 2 alkyl, C2-C12 alkenyl, Ci-Ci2 alkyl esters, C? -C? 2 alkyl amides, C4-C12 alkenyl esters, C? -C? 2 alkylaryl esters , C4-C12 alkenylaryl esters, C4-C12 alkenyl amides, C? -C12 alkoxy, Cx-C12 alkyl formyl, C2-C12 formyl alkenyl, C? -C? 2 alkanoyloxy, C2-C12 alkanoyl alkenyl, carboxy alkyl C? -C? 2, C2-C12 carboxy alkenyl, wherein the alkyl and alkenyl groups are optionally substituted with 1-6 substituent groups selected from the group consisting of: C3-C? 2 cycloalkyl, C3-C12 heterocyclyl, aryl, heteroaryl, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, halogen, C 2 -C 6 alkenyl, OH, nucleosides, nucleotides, purines, pyrimidines, aromatic esters, aryl esters, cycloalkyl esters, cycloalkenyl esters, purines, or pyrimidines; OR? is OH or -0 (CO) CH3; R 2 is -CH = CHCH = CH 2, -CH 2 -phenyl, or -CH 2 -pyridyl, wherein the phenyl and pyridyl groups are optionally substituted at each substitutable position with a group that is independently C 1 -C 6 alkyl, C 1 alkoxy C6, haloalkyl, haloalkoxy, hydroxy, hydroxyalkyl, halogen, -C02H, C6 -C6 alkoxycarbonyl, -C (0) NH2, -C (O) NH (C? -C6 alkyl), or -C (0) N ( C? -C6 alkyl) (C? -C6 alkyl) or pharmaceutically acceptable salts, solvates, esters, amides, hydrates, or combinations thereof.

2. The compound according to claim 1, characterized in that R is wherein Hal is chloro, fluoro, iodo, or bromine; R3 is selected from H, OH, NH2, halogen, and N02, and Y is selected from CH, N, O, and S.

3. The compound according to claim 1, characterized in that the compound is selected from the group consisting of: . Y

4. A pharmaceutical composition characterized in that it comprises at least one compound, pharmaceutically acceptable salt, solvate, hydrate, ester, amide, or isomer according to claim 1, and at least one pharmaceutically acceptable carrier, excipient, solvent, adjuvant, diluent, or mixtures thereof .

5. A pharmaceutical composition according to claim 4, characterized in that the pharmaceutical composition is in unit dosage form.

6. The pharmaceutical composition in unit dosage form according to claim 5, characterized in that the compound of Formula I is present in an amount of about 1 pg to about 100 mg.

7. A method of treatment for mucociliary dysfunction in a subject characterized in that it comprises administering to a subject in need of such treatment a therapeutically effective amount of a compound according to claim 1, or salts, solvates, hydrates, esters, amides, or pharmaceutically acceptable isomers thereof.

A method of treatment for mucociliary dysfunction in a subject characterized in that it comprises administering to a subject in need of such treatment a therapeutically effective amount of a pharmaceutical composition in accordance with claim 4.

9. A method of treatment, prevention, or treatment and prevention of diseases associated with decreased mucus clearance in a mammal characterized in that it comprises administering to a mammal in need of such treatment, a therapeutically effective amount of a compound according to claim 1 or salts, solvates, hydrates, esters, amides , or pharmaceutically acceptable isomers thereof.

A method of treatment, prevention, or treatment and prevention of diseases associated with decreased mucus clearance in a mammal characterized in that it comprises administering to a mammal in need of such treatment a therapeutically effective amount of a pharmaceutical composition in accordance with claim 4

11. A method of treating diseases associated with decreased purification of mucus in a mammal, characterized in that it comprises administering to a mammal in need of such treatment a therapeutically effective amount of a compound according to claim 1 or pharmaceutically acceptable salts, solvates, hydrates, esters, amides, or isomers thereof.

12. A method of treating diseases associated with decreased clearance of mucus in a mammal, characterized in that it comprises administering to a mammal in need of such treatment a therapeutically effective amount of a pharmaceutical composition in accordance with claim 4.

13. The method of according to claim 12, characterized in that the therapeutically effective amount is administered in a dose of between about 0.1 pg to about 100 mg per day.

The method according to claim 12, characterized in that the therapeutically effective amount comprises a dose of between about 0.1 mg to about 10 mg per day.

15. A method of treating a subject who has, or is at risk of increasing, the development of chronic obstructive pulmonary disease, asthma, lung disease, lung infection, or cystic fibrosis, characterized in that it comprises administering to a subject in need of such treating a therapeutically effective amount of a compound according to claim 1 or pharmaceutically acceptable salts, solvates, hydrates, esters, amides, or isomers thereof.

16. A method of treating a subject who has, or is at risk of increasing, the development of chronic obstructive pulmonary disease, asthma, lung disease, lung infection, or cystic fibrosis, characterized in that it comprises administering to a subject in need of such treating a therapeutically effective amount of a pharmaceutical composition of the claim.

17. A method for regulating the clearance of mucus in a cell, characterized in that it comprises contacting a cell with an effective amount of a compound according to claim 1 or pharmaceutically acceptable salts, solvates, hydrates, esters, amides, or isomers of the same.

18. A method for regulating the clearance of mucus in a cell, characterized in that it comprises contacting a cell with an effective amount of a pharmaceutical composition according to claim 4.

19. A compound according to claim 1, characterized in that R e R2 is

20. The compound according to claim 19, characterized in that Rx is H.

The compound according to claim 2, characterized in that R2 is -CH = CHCH = CH2.

22. A method of treating an industrial disease characterized in that it comprises administering a pharmaceutically acceptable amount of a compound according to claim 1, or pharmaceutically acceptable salts, solvates, hydrates, esters, amides, or isomers thereof, to a subject that You need such treatment.

23. A method of treatment according to claim 22, characterized in that the industrial disease is caused or aggravated by inhalation of gases, textile particles, sand, or other industrial particles or fumes.

24. A method of treating a disease or condition resulting from inhalation of bacteria or other pathogenic particles characterized in that it comprises administering a pharmaceutically acceptable amount of a compound according to claim 1, or salts, solvates, hydrates, esters, amides, or pharmaceutically acceptable isomers thereof, to a subject in need of such treatment.