AU1598799A - Use of substituted 11-phenyl-dibenzazepine compounds for the treatment or prevention of sickle cell disease, inflammatory diseases characterized by abnormal cell proliferation, diarrhea and scour - Google Patents

Description

WO 99/26628 PCT/US98/24967 -1 USE OF SUBSTITUTED 11-PHENYL-DIBENZAZEPINE COMPOUNDS FOR THE TREATMENT OR PREVENTION OF SICKLE CELL DISEASE, INFLAMMATORY DISEASES CHARACTERIZED BY ABNORMAL CELL PROLIFERATION, DIARRHEA AND SCOUR 5 Field of the Invention The present invention relates to aromatic organic compounds which are specific, potent and safe inhibitors of the Ca 2 +-activated potassium channel (Gardos channel) of erythrocytes, of mammalian cell proliferation, and/or of secretagogue-stimulated o transepithelial electrogenic chloride secretion in intestinal cells. The compounds are generally substituted 11-phenyl-dibenzazepine compounds. The compounds can be used to reduce sickle erythrocyte dehydration and/or delay the occurrence of erythrocyte sickling or deformation in situ as a therapeutic approach towards the treatment or prevention of sickle cell disease. The compounds can also be used to inhibit mammalian cell proliferation in situ as a 15 therapeutic approach towards the treatment or prevention of diseases characterized by abnormal cell proliferation. Furthermore, the compounds can be used to inhibit chloride secretion as a therapeutic approach towards the treatment of diarrhea and scours. Background of the Invention 20 Sickle cell disease has been recognized within West Africa for several centuries. Sickle cell anemia and the existence of sickle hemoglobin (Hb S) was the first genetic disease to be understood at the molecular level. It is recognized today as the morphological and clinical result of a glycine to valine substitution at the No. 6 position of the beta globin chain (Ingram, 1956, Nature 178:792-794). The origin of the amino acid change and of the disease 25 state is the consequence of a single nucleotide substitution (Marotta et al., 1977, J. Biol. Chem. 252:5040-5053). The major source of morbidity and mortality of patients suffering from sickle cell disease is vascular occlusion caused by the sickled cells, which causes repeated episodes of pain in both acute and chronic form and also causes ongoing organ damage with the passage 30 of time. It has long been recognized and accepted that the deformation and distortion of sickle cell erythrocytes upon complete deoxygenation is caused by polymerization and intracellular gelation of sickle hemoglobin, hemoglobin S (Hb S). The phenomenon is well reviewed and discussed by Eaton and Hofrichter, 1987, Blood 70:1245. The intracellular gelatin and WO 99/26628 PCT/US98/24967 -2 polymerization of Hb S can occur at any time during erythrocyte's journey through the vasculature. Thus, erythrocytes in patients with sickle cell disease containing no polymerized hemoglobin S may pass through the microcirculation and return to the lungs without sickling, may sickle in the veins or may sickle in the capillaries. 5 The probability of each of these events is determined by the delay time for intracellular gelation relative to the appropriate capillary transit time (Eaton et al., 1976, Blood 47:621). In turn, the delay time is dependent upon the oxygenation state of the hemoglobin, with deoxygenation shortening the delay time. Thus, if it is thermodynamically impossible for intracellular gelation to take place, or if the delay time at venous oxygen 10 pressures is longer than about 15 seconds, cell sickling will not occur. Alternatively, if the delay time is between about 1 and 15 seconds, the red cell will likely sickle in the veins. However, if the delay time is less than about 1 second, red cells will sickle within the capillaries. For red cells that sickle within the capillaries, a number of possible consequent events 15 exist, ranging from no effect on transit time, to transient occlusion of the capillary, to a more permanent blockage that may ultimately result in ischemia or infarction of the surrounding cells, and in destruction of the red cell. It has long been recognized that the cytoplasm of the normal erythrocyte comprises approximately 70% water. Water crosses a normal erythrocyte membrane in milliseconds; 20 however, the loss of cell water causes an exponential increase in cytoplasmic viscosity as the mean cell hemoglobin concentration (MCHC) rises above about 32 g/dl. Since cytoplasmic viscosity is a major determinate of erythrocyte deformability and sickling, the dehydration of the erythrocyte has substantial rheological and pathological consequences. Thus, the physiological mechanisms that maintain the water content of a normal erythrocytes and the 25 pathological conditions that cause loss of water from erythrocytes in the blood circulation are critically important. Not surprisingly, regulation of erythrocyte dehydration has been recognized as an important therapeutic approach towards the treatment of sickle cell disease. Since cell water will follow any osmotic change in the intracellular concentration of ions, the maintenance of the red cell's potassium concentration is of particular importance (Stuart and 30 Ellory, 1988, Brit J. Haematol. 69:1-4). Many attempts and approaches to therapeutically treating dehydrated sickle cells (and thus decreasing polymerization of hemoglobin S by lowering the osmolality of plasma) have WO 99/26628 PCT/US98/24967 -3 been tried with limited success, including the following approaches: intravenous infusion of distilled water (Gye et al., 1973, Am. J. Med. Sci. 266:267-277); administration of the antidiuretic hormone vasopressin together with a high fluid intake and salt restriction (Rosa et al., 1980, M. Eng. J. Med. 303:1138-1143; Charache and Walker, 1981, Blood 58:892-896); 5 the use of monensin to increase the cation content of the sickle cell (Clark et al., 1982, J. Clin. Invest. 70:1074-1080; Fahim and Pressman, 1981, Life Sciences 29:1959-1966); intravenous administration of cetiedil citrate (Benjamin et al., 1986, Blood 67:1442-1447; Berkowitz and Orringer, 1984, Am. J. Hematol. 17:217-223; Stuart et al., 1987, J. Clin. Pathol. 40:1182-1186); and the use of oxpentifylline (Stuart et al., 1987, J. Clin. Pathol. 10 40:1182-1186). Another approach towards therapeutically treating dehydrated sickle cells involves the administration of imidazole, nitroimidazole and triazole antimycotic agents such as Clotrimazole (U.S. Patent No. 5,273,992 to Brugnara et al.). Clotrimazole, an imidazole containing antimycotic agent, has been shown to be a specific, potent inhibitor of the Gardos 15 channel of normal and sickle erythrocytes, and prevents Ca2+-dependent dehydration of sickle cells both in vitro and in vivo (Brugnara et al., 1993, J. Clin. Invest. 92:520-526; De Franceschi et al., 1994, J. Clin. Invest. 93:1670-1676). When combined with a compound which stabilizes the oxyconformation of Hb S, Clotrimazole induces an additive reduction in the clogging rate of a micropore filter and may attenuate the formation of irreversibly sickled 20 cells (Stuart et al., 1994, J. Haematol. 86:820-823). Other compounds that contain a heteroaryl imidazole-like moiety believed to be useful in reducing sickle erythrocyte dehydration via Gardos channel inhibition include miconazole, econazole, butoconazole, oxiconazole and sulconazole. Each of these compounds is a known antimycotic. Other imidazole-containing compounds have been found to be incapable of inhibiting the Gardos 25 channel and preventing loss of potassium. As can be seen from the above discussion, reducing sickle erythrocyte dehydration via blockade of the Gardos channel is a powerful therapeutic approach towards the treatment and/or prevention of sickle cell disease. Compounds capable of inhibiting the Gardos channel as a means of reducing sickle cell dehydration are highly desirable, and are therefore an object 30 of the present invention. Cell proliferation is a normal part of mammalian existence, necessary for life itself. However, cell proliferation is not always desirable, and has recently been shown to be the root WO 99/26628 PCT/US98/24967 -4 of many life-threatening diseases such as cancer, certain skin disorders, inflammatory diseases, fibrotic conditions and arteriosclerotic conditions. Cell proliferation is critically dependent on the regulated movement of ions across various cellular compartments, and is associated with the synthesis of DNA. Binding of 5 specific polypeptide growth factors to specific receptors in growth-arrested cells triggers an array of early ionic signals that are critical in the cascade of mitogenic events eventually leading to DNA synthesis (Rozengurt, 1986, Science 234:161-164). These include (1) a rapid increase in cystolic Ca 2 + , mostly due to rapid release of Ca 2+ from intracellular stores; (2) capacitative Ca 2+ influx in response to opening of ligand-bound and hyperpolarization 10 sensitive Ca 2+ channels in the plasma membrane that contribute further to increased intracellular Ca 2+ concentration (Tsien and Tsien, 1990, Annu. Rev. Cell Biol. 6:715-760; Peppelenbosch et al., 1991, J. Biol. Chem. 266:19938-19944); and (3) activation of Ca 2+ dependent K' channels in the plasma membrane with increased K' conductance and membrane hyperpolarization (Magni et al., 1991, J. Biol. Chem. 261:9321-9327). These 15 mitogen-induced early ionic changes, considered critical events in the signal transduction pathways, are powerful therapeutic targets for inhibition of cell proliferation in normal and malignant cells. One therapeutic approach towards the treatment of diseases characterized by unwanted or abnormal cell proliferation via alteration of the ionic fluxes associated with early mitogenic 20 signals involves the administration of Clotrimazole. Clotrimazole has been shown to inhibit the Ca 2 +-activated potassium channel of erythrocytes. In addition, Clotrimazole inhibits voltage- and ligand-stimulated Ca 2 + influx mechanisms in nucleated cells (Villalobos et al., 1992, FASEB J. 6:2742-2747; Montero et al., 1991, Biochem. J. 277:73-79) and inhibits cell proliferation both in vitro and in vivo (Benzaquen et al., 1995, Nature Medicine 1:534-540). 25 Recently, Clotrimazole and other imidazole-containing antimycotic agents capable of inhibiting Ca 2 +-activated potassium channels have been shown to be useful in the treatment of arteriosclerosis (U.S. Patent No. 5,358,959 to Halperin et al.), as well as other disorders characterized by unwanted or abnormal cell proliferation. As can be seen from the above discussion, inhibiting mammalian cell proliferation via 30 alteration of ionic fluxes associated with early mitogenic signals is a powerful therapeutic approach towards the treatment and/or prevention of diseases characterized by unwanted or abnormal cell proliferation. Compounds capable of inhibiting mammalian cell proliferation WO 99/26628 PCT/US98/24967 -5 are highly desirable, and are therefore also an object of the present invention. Acute and chronic diarrheas represent a major medical problem in many areas of the world. Diarrhea is both a significant factor in malnutrition and the leading cause of death (5,000,000 deaths/year) in children less than five years old. Secretory diarrheas are also a 5 dangerous condition in patients of acquired immunodeficiency syndrome (AIDS) and chronic inflammatory bowel disease (IBD). 16 million travelers to developing countries from industrialized nations every year develop diarrhea, with the severity and number of cases of diarrhea varying depending on the country and area of travel. The major medical consequences of diarrheal diseases include dehydration, acidosis, death and impaired growth. 10 Diarrhea in barn animals and pets such as cows, pigs and horses, sheep, goats, cats and dogs, also known as scours, is a major cause of death in these animals. Diarrhea can result from any major transition, such as weaning or physical movement. One form of diarrhea is characterized by diarrhea in response to a bacterial or viral infection and generally occurs within the first few hours of the animal's life. 15 Although the major consequences of diarrheal diseases are very similar, there are numerous causes of diarrhea. Secretory and exudative diarrhea are primarily caused by bacterial or viral infections. The most common diarrheal causing bacteria is enterotoxogenic E-coli (ETEC) having the K99 pilus antigen. Common viral causes of diarrhea include rotavirus and coronavirus. Other infectious agents include cryptosporidium, giardia lamblia, 20 and salmonella, among others. The treatment for diarrhea depends on the patient and the infection source. Diarrhea which is found in travelers to industrialized nations (travelers diarrhea) frequently is caused by bacterial pathogens which are acquired through ingestion of fecally contaminated food and/or water. Approximately 50-75% of these cases are attributed to ETEC. Although 25 traveler's diarrhea is painful, it is generally not life-threatening and often the symptoms last only 3-5 days. The symptoms include urgent diarrhea, abdominal cramps, nausea and fever. The most effective course of treatment for traveler's diarrhea is the administration of antibiotics in conjunction with oral rehydration. It has been shown that prophylactic administration of antibiotics drastically reduces the number of travelers experiencing 30 symptoms of diarrhea. However, routine administration of antibiotics is not suggested as it may cause resistant strains of a bacteria to develop. Other treatment methods include administration of bismuth subsalicylate, often taken in the form of Pepto-Bismal, WO 99/26628 PCT/US98/24967 -6 diphenoxylate and loperamide. Diarrhea in AIDS patients is a very serious condition which causes wasting and may be an important factor in the decline of these patients. AIDS patients often develop diarrhea due to enteric infections which their immune system is not capable of fighting off, but AIDS 5 patients may also develop diarrhea by AIDS enteropathy. AIDS enteropathy is a disorder characterized by diarrhea without the involvement of secondary infections. It is caused by the human immunodeficiency virus (HIV) infection of the small bowel mucosal cells and colonic mucosal cells. The most common infective agent causing diarrhea due to enteric infection in AIDS patients in cryptosporidium. The methods for treating diarrhea in AIDS patients 10 include administration of antibiotics and administration of immunoglobulins or an immunoglobulin enriched fraction of bovine colostrum. Colostrum, which is the first milk produced by mammals after birthing is enriched with antibodies. Acute diarrhea or scours, is a main cause of death in many newborn barn animals such as calves and pigs. Scours is often caused by ETEC with a K99 pilus antigen. Infection with 15 the ETEC causes hypersecretion of fluid and electrolytes. Hypersecretion in turn causes dehydration and pH imbalance which may result in death of the newborn calf or pig. Newborn barn animals are also susceptible to viral infectious agents causing scours. Infections with rotavirus and coronavirus are common in newborn calves and pigs. Rotavirus infection often occurs within 12 hours of birth. Symptoms of rotaviral infection include 20 excretion of watery feces, dehydration and weakness. Coronavirus which causes a more severe illness in the newborn animals, has a higher mortality rate than rotaviral infection. Often, however, a young animal may be infected with more than one virus or with a combination of viral and bacterial microorganisms at one time. This dramatically increases the severity of the disease. 25 Generally the best protection for a newborn barn animal from viral or bacterial infection is the consumption of colostrum. If the mother animal has been exposed to these infectious agents then the colostrum will contain antibodies, which are often sufficient to protect the newborn from contracting the diseases. Sometimes, however, this is not sufficient and the animals need further protection. A common method of treatment includes 30 administration of a concentrated colostrum solution or an immunoglobulin fraction isolated from a colostrum solution. This oral treatment may be combined with rehydration salts. Although these methods have improved the morbidity and mortality rate of newborn animals WO 99/26628 PCT/US98/24967 -7 having scours, there still exists a need for more effective treatments. Certain imidazoles such as clotrimazole are agents which have been used both topically and systemically as antifungals. More recently, studies have identified other uses for such imidazoles. U.S. patent no. 5,273,992 revealed that these imidazoles regulate Ca" 5 activated K' channels in erythrocytes, and are thus useful in treating sickle cell anemia, which involves the inhibition of potassium transport. These imidazoles have also been found to be effective in inhibiting endothelial and/or vascular smooth muscle cell proliferation. The results of this finding are described in U.S. patent no. 5,358,959 and U.S. serial no. 08/018,840, which discloses using clotrimazole for treating atherosclerotic and angiogenic 10 conditions, respectively. Nonimidazole metabolites and analogs of the foregoing compounds also have been described as useful in treating the foregoing conditions (see U.S. serials nos. 08/307,874 and 08/307,887). Summary of the Invention 15 These and other objects are provided by the present invention, which in one aspect provides a class of organic compounds which are potent, selective and safe inhibitors of the Ca2+-activated potassium channel (Gardos channel) of erythrocytes, of mammalian cell proliferation and/or of secretagogue-stimulated transepithelial electrogenic chloride secretion in intestinal cells. The compounds can be used to reduce sickle erythrocyte dehydration 20 and/or delay the occurrence of erythrocyte sickling or deformation in situ as a therapeutic approach towards the treatment or prevention of sickle cell disease. The compounds can also be used to inhibit mammalian cell proliferation in situ as a therapeutic approach towards the treatment or prevention of diseases characterized by abnormal cell proliferation. Furthermore, the compounds can also be used to inhibit chloride secretion in intestinal cells as a therapeutic 25 approach towards the treatment of diarrhea and scours. The compounds are generally substituted 11-phenyl-dibenzazepine compounds. In one illustrative embodiment, the compounds capable of inhibiting the Gardos channel, mammalian cell proliferation and/or secretagogue-stimulated transepithelial electrogenic chloride secretion in intestinal cells according to the invention, are compounds having the structural formula (I): 30 WO 99/26628 PCT/US98/24967 -8 (I)

R

15

R

6 Rs N R7 R4 R8 R9 R R2 R3 R°1 R 1 4 R11 R13 R12 or pharmaceutically acceptable salts of hydrates thereof, wherein: R, is -R', (C 6

-C

20 ) aryl or substituted (C 6

-C

2 0 ) aryl;

R

2 is -R', -OR', -SR', halogen or trihalomethyl; 5 R 3 is -R', -OR', -SR', halogen or trihalomethyl or, when taken together with R 4 , is (C 6 C 20 ) aryleno;

R

4 is -R', -OR', -SR', halogen or trihalomethyl or, when taken together with R 3 , is (C 6 C 20 ) aryleno; each of R 5 , R 6 , R 7 , R 8 , R, R 1 0 , R 11 , R 2 , R 1 3 and R 14 is independently selected from the 10 group consisting of -R', halogen and trihalomethyl; Rs is -R", -C(O)R", -C(S)R", -C(O)OR", -C(S)OR", -C(O)SR", -C(S)SR",

-C(O)N(R")

2 , -C(S)N(R") 2 , -C(O)C(O)R", -C(S)C(O)R", -C(O)C(S)R", -C(S)C(S)R", -C(O)C(O)OR", -C(S)C(O)OR", -C(O)C(S)OR", -C(O)C(O)SR", -C(S)C(S)OR", -C(S)C(O)SR", -C(O)C(S)SR", -C(S)C(S)SR", -C(O)C(O)N(R") 2 , -C(S)C(O)N(R") 2 , 15 -C(O)C(S)N(R") 2 or-C(S)C(S)N(R")2 each R' is independently selected from the group consisting of -H, (C-C 6 ) alkyl,

(C

1

-C

6 ) alkenyl and (Cl-C 6 ) alkynyl; each R" is independently selected from the group consisting of -H, (C 1

-C

6 ) alkyl,

(C

1

-C

6 ) alkenyl, (C 1

-C

6 ) alkynyl, (C 6

-C

20 ) aryl, substituted (C 6

-C

2 0 ) aryl, (C 6

-C

2 6 ) alkaryl and 20 substituted (C 6

-C

26 ) alkaryl; and the aryl and alkaryl substituents are each independently selected from the group consisting of -CN, -OR', -SR', -NO 2 , -NR'R', halogen, (C 1

-C

6 ) alkyl, (C 1

-C

6 ) alkenyl, (C 1

-C

6

)

WO 99/26628 PCT/US98/24967 -9 alkynyl and trihalomethyl. In a preferred embodiment of the invention, the chalcogens in the compounds of formula (I) are each oxygen. In another preferred embodiment, the compounds are those of structure (I) wherein the 5 halogens are each independently -F, -Cl, -Br or -I. In another preferred embodiment, the alkyl, alkenyl and alkynyl groups are each independently (C 1

-C

3 ) and/or the aryl groups are phenyl and/or the aryleno groups are benzeno. In another preferred embodiment, Rs, R 6 , R 7 , R 9 , Rio, R I and R, 3 are each 10 independently -R'. In another preferred embodiment, the substituted aryl and alkaryl are mono substituted. In another preferred embodiment, R 1 5 is -R", -C(O)R", -C(O)OR", -C(O)N(R") 2 , -C(O)C(O)R", -C(O)C(O)OR" or -C(O)C(O)N(R") 2 . 15 In another preferred embodiment of the invention, the compounds are those of structural formula (I) wherein: R, is -R' or (C 6

-C

20 ) aryl; R 2 is -R' or -OR'; R 3 is -R' or -OR' or, when taken together with R 4 , is (C 6

-C

20 ) aryleno; R 4 is -R' or -OR' or, when taken together with R 3 , is (C 6

-C

20 ) aryleno; each of Rs, R 6 , R 7 , R 8 , R 9 , Rio, RN 1 , R 1 2 , RB 3 and R 14 is independently selected from the group consisting of-R' and halogen; R 5 is -R", -C(O)R", 20 -C(O)OR", -C(O)N(R") 2 , -C(O)C(O)R", -C(O)C(O)OR" or -C(O)C(O)N(R") 2 ; each R' is independently selected from the group consisting of-H, (CI-C 6 ) alkyl, (CI-C 6 ) alkenyl and

(C

1

-C

6 ) alkynyl; each R" is independently selected from the group consisting of -H, (C 1

-C

6 ) alkyl, (CI-C 6 ) alkenyl, (C 1

-C

6 ) alkynyl, (C 6

-C

20 ) aryl, substituted (C 6

-C

20 ) aryl, (C 6

-C

26 ) alkaryl and substituted (C 6

-C

26 ) alkaryl; and the aryl and alkaryl substituents are each independently 25 selected from the group consisting of -CN, -OR', -NO 2 , -NR'R', halogen, (C 1

-C

6 ) alkyl,

(C

1

-C

6 ) alkenyl and (C 1

-C

6 ) alkynyl. In another preferred embodiment, the compounds are those of formula (I) wherein: R, is -R' or (C 6

-C

10 ) aryl; R 2 is -R' or -OR'; R 3 is -R' or -OR' or, when taken together with R 14 , is

(C

6

-CI

0 ) aryleno; R 4 is -R' or -OR' or, when taken together with R 3 , is (C 6

-C

10 ) aryleno; each of 30 R 5 , R 6 and R 7 is -H; R 8 is -R', -F, -Cl, -Br or -I; each of R 9 , RIo and Rl is -H; R, 2 is -R', -F, -Cl, -Br or -I; R 1 3 is -H; R 1 4 is -R', -F, -Cl, -Br or -I; R 15 is -R", -C(O)R", -C(O)OR", -C(O)N(R")2, -C(O)C(O)R", -C(O)C(O)OR" or -C(O)C(O)N(R") 2 ; each R' is independently WO 99/26628 PCT/US98/24967 - 10 selected from the group consisting of -H, (C

,

-C

3 ) alkyl, (Ci-C 3 ) alkenyl and (C 1

-C

3 ) alkynyl; each R" is independently selected from the group consisting of -H, (C -C 3 ) alkyl, (CI-C 3 ) alkenyl, (C]-C 3 ) alkynyl, (C 6

-C

1 0 ) aryl, substituted (C 6

-C

1 0 ) aryl, (C 6

-C

13 ) alkaryl or substituted

(C

6

-C

1 3 ) alkaryl; and the aryl and alkaryl substituents are each independently selected from the 5 group consisting of -OR',-NO 2 , -NR'R', -F, -Cl, -Br, -I, (C 1

-C

3 ) alkyl, (C 1

-C

3 ) alkenyl and

(C

1

-C

3 ) alkynyl. In still another preferred embodiment, the compounds are those of structural formula (I) wherein: R, is -R' or phenyl; R 2 is -R' or -OR'; R 3 is -R' or -OR' or, when taken together with R 4 , is benzeno; R 4 is -R' or -OR' or, when taken together with R 3 , is benzeno; each of Rs, 10 R 6 and R 7 is -H; R 8 is -R', -Cl or -Br; each of R 9 , RIo and R,, is -H; RI 2 is -R', -F or -Cl; R 1 3 is -H; RI 4 is -R' or -Cl; R 15 is -R", -C(O)R", -C(O)OR", -C(O)NHR", -C(O)C(O)R" or -C(O)C(O)OR"; each R' is independently selected from the group consisting of-H, (CI-C 3 ) alkyl, (CI-C 3 ) alkenyl and (CI-C 3 ) alkynyl; each R" is independently selected from the group consisting of-H, (Ci-C 3 ) alkyl, (C 1

-C

3 ) alkenyl, (C 1

-C

3 ) alkynyl, (C 6 -Ci 0 ) aryl, mono 15 substituted (C 6

-C

1 0 ) aryl, (C 6

-C

13 ) alkaryl or mono-substituted (C 6

-C

13 ) alkaryl; and the aryl and alkaryl substituents are each independently selected from the group consisting of -OR',

-NO

2 , -NR'R', -Cl, (C 1

-C

3 ) alkyl, (C 1

-C

3 ) alkenyl and (C 1

-C

3 )alkynyl. In still another aspect, the invention provides a method for reducing sickle erythrocyte dehydration and/or delaying the occurrence of erythrocyte sickling or deformation in situ. 20 The method involves contacting a sickle erythrocyte in situ with an amount of at least one compound according to the invention, or a pharmaceutical composition thereof, effective to reduce sickle erythrocyte dehydration and/or delay the occurrence of erythrocyte sickling or deformation. In a preferred embodiment, the sickle cell dehydration is reduced and erythrocyte deformation is delayed in a sickle erythrocyte that is within the microcirculation 25 vasculature of a subject, thereby preventing or reducing the vaso-occlusion and consequent adverse effects that are commonly caused by sickled cells. In still another aspect, the invention provides a method for the treatment and/or prevention of sickle cell disease in a subject, such as a human. The method involves administering a prophylactically or therapeutically effective amount of at least one compound 30 according to the invention, or a pharmaceutical composition thereof, to a patient suffering from sickle cell disease. The patient may be suffering from either acute sickle crisis or chronic sickle cell episodes.

WO 99/26628 PCT/US98/24967 -11 In one aspect of the invention a method is provided for inhibiting unwanted cellular proliferation associated with an inflammatory disease. The method includes the step of contacting a cell the proliferation of which contributes to inflammation in situ with an amount of a compound having the above described formula (I) effective to inhibit proliferation of the 5 cell. In one embodiment the method of administration is selected from the group consisting of oral, parenteral, intravenous, subcutaneous, transdermal and transmucosal for a living human. In one embodiment the mammalian cell is a fibrotic cell or a lymphocyte. According to another aspect of the invention a method is provided for treating or preventing an inflammatory disease. The method includes the step of administering to a 10 subject in need of such treatment a therapeutically effective amount of a compound of the above-described formula (I). In one embodiment the inflammatory disease is diarrhea. Preferably the diarrhea is caused by inflammatory bowel disease. In another embodiment the inflammatory disease is an autoimmune disease. In other embodiments the inflammatory disease is selected from the group consisting of proliferative glomerulonephritis; lupus 15 erythematosus; scleroderma; temporal arteritis; thromboangiitis obliterans; mucocutaneous lymph node syndrome; asthma; host versus graft; inflammatory bowel disease; multiple sclerosis; rheumatoid arthritis; thyroiditis; Grave's disease; antigen-induced airway hyperactivity; pulmonary eosinophilia; Guillain-Barre syndrome; allergic rhinitis; myasthenia gravis; human T-lymphotrophic virus type 1-associated myelopathy; herpes simplex 20 encephalitis; inflammatory myopathies; atherosclerosis; and Goodpasture's syndrome. In certain embodiments the administration is parenteral or per oral. In yet another aspect, the invention provides a method for inhibiting mammalian cell proliferation in situ. Preferably, the mammalian cell proliferation is not associated with a proliferative disease selected from the group consisting of cancer, actinic keratosis, and 25 Kaposi's sarcoma. The method involves contacting a mammalian cell in situ with an amount of at least one compound according to the invention, or a pharmaceutical composition thereof, effective to inhibit cell proliferation. The compound or composition may act either cytostatically, cytotoxically or a by a combination of both mechanisms to inhibit proliferation. Mammalian cells in this manner include vascular smooth muscle cells, fibroblasts and 30 endothelial cells. In still another aspect, the invention provides a method for treating and/or preventing unwanted or abnormal cell proliferation in a subject, such as a human. Preferably, the WO 99/26628 PCT/US98/24967 -12 unwanted or abnormal cell proliferation is not associated with a proliferative disease selected from the group consisting of cancer, actinic keratosis, and Kaposi's sarcoma. In the method, at least one compound according to the invention, or a pharmaceutical composition thereof, is administered to a subject in need of such treatment in an amount effective to inhibit the 5 unwanted or abnormal mammalian cell proliferation. The compound and/or composition may be applied locally to the proliferating cells, or may be administered to the subject systemically. Preferably, the compound and/or composition is administered to a subject that has a disorder characterized by unwanted or abnormal cell proliferation, and preferably the unwanted or abnormal cell proliferation is not associated with a proliferative disease selected 10 from the group consisting of cancer, actinic keratosis, and Kaposi's sarcoma. Such disorders include, but are not limited to, non-cancerous angiogenic conditions or arteriosclerosis. In yet another aspect, the invention provides a method for the treatment and/or prevention of diseases that are characterized by unwanted and/or abnormal mammalian cell proliferation. Preferably, the unwanted or abnormal cell proliferation is not associated with a 15 proliferative disease selected from the group consisting of cancer, actinic keratosis, and Kaposi's sarcoma. The method involves administering a prophylactically or therapeutically effective amount of at least one compound according to the invention, or a pharmaceutical composition thereof, to a subject in need of such treatment. Diseases that are characterized by abnormal mammalian cell proliferation which can be treated or prevented by way of the 20 methods of the invention include, but are not limited to, blood vessel proliferative disorders, fibrotic disorders and arteriosclerotic conditions. According to another aspect of the invention, a method for treating diarrhea of diverse etiology is provided. The method involves administering to a subject who is in need of such treatment, an aromatic compound of the invention in an amount effective to inhibit the 25 diarrhea. Preferably the compound is administered orally in conjunction with oral rehydration fluids. The aromatic compounds useful in the invention are substituted 11-phenyl dibenzazepine, or analogues thereof. In one illustrative embodiment, the aromatic compounds according to the invention are compounds having the above-described formula (I). According to one embodiment of the invention the subject in need of such treatment is a subject who has 30 symptoms of diarrhea or scours. In another embodiment of the invention, the subject in need of such treatment is a subject at risk of developing diarrhea or scours. In general diarrhea is a secretory disorder, which is caused by at least one of several WO 99/26628 PCT/US98/24967 - 13 mechanisms. In one embodiment the diarrhea is an exudative form of diarrhea; In one embodiment the diarrhea is a nonexudative form of diarrhea; In another embodiment the diarrhea is a decreased absorption form of diarrhea; In another embodiment the diarrhea is a non-decreased absorption form of diarrhea; In yet another embodiment the diarrhea is a 5 secretory form of diarrhea. In yet another embodiment the diarrhea is a nonsecretory form of diarrhea. In still another embodiment the diarrhea is a noninflammatory form of diarrhea. In another aspect, the present invention provides pharmaceutical compositions comprising one or more compounds according to the invention in admixture with a pharmaceutically acceptable carrier, excipient or diluent. Such a preparation can be 10 administered in the methods of the invention. According to another aspect of the invention, pharmaceutical preparations are provided, comprising one or more of the aromatic compounds of the invention in admixture with a pharmaceutically acceptable carrier, excipient or diluent, wherein the aromatic compound(s) of the invention is (are) in an amount effective for treating: (i) unwanted or 15 abnormal cell proliferation, preferably not a proliferative disease selected from the group consisting of cancer, actinic keratosis, and Kaposi's sarcoma; (ii) an inflammatory disease; (iii) sickle cell disease; and (iv) diarrhea or scours. In one embodiment, the aromatic compounds useful according to the invention have the general formula (I) provided above. In certain other embodiments, the pharmaceutical preparations include the aromatic compounds 20 of the invention together with a non-formula (I) agent selected from the group consisting of an anti-proliferative agent; (ii) an anti-inflammatory agent; (iii) anti-sickle cell agent; and (iv) an anti-diarrhea or anti-scours agent. According to another aspect, the use of aromatic compounds of the invention in the manufacture of medicaments is provided. The medicaments are useful for treating: (i) 25 unwanted or abnormal cell proliferation, preferably not a proliferative disease that includes cancer, actinic keratosis, and Kaposi's sarcoma; (ii) an inflammatory disease; (iii) sickle cell disease; and (iv) diarrhea or scours. According to another aspect of the invention, pharmaceutical preparations are provided. These pharmaceutical preparations include the aromatic compounds of the 30 invention together with an anti-diarrheal agent. In one embodiment, the aromatic compounds useful according to the invention have the general formula (I) provided above. In other embodiments the aromatic compounds useful according to the invention are the preferred WO 99/26628 PCT/US98/24967 - 14 compounds described above. Preferably the pharmaceutical, composition of the invention may be administered orally. The invention also provides the aromatic compounds of the invention in the manufacture of a medicament for the treatment of diarrhea or scours. In one embodiment, the 5 aromatic compounds useful according to the invention have the general formula (I) provided above. In other embodiments the aromatic compounds useful according to the invention are the preferred compounds described above. According to another aspect of the invention, veterinary preparations are provided. These veterinary preparations include the aromatic compounds useful according to the 10 invention together with an anti-scours preparation. In one embodiment, the aromatic compounds useful according to the invention have the general formula (I) provided above. In other embodiments the aromatic compounds useful according to the invention are the preferred compounds described above. Each of the limitations of the invention can encompass various embodiments of the 15 invention. It is, therefore, anticipated that each of the limitations of the invention involving any one element or combinations of elements can be included in each aspect of the invention. Brief Description of the Drawings 20 FIG. 1 is a general reaction scheme for synthesizing certain compounds according to the invention; FIG. 2 is a general reaction scheme for synthesizing certain compounds according to the invention; FIG. 3 is a bar graph depicting the effect of clotrimazole in the inhibition of cAMP and 25 Ca" dependent Cl- secretion in T84 cells; and FIG. 4 is a graph showing the effect of clotrimazole on the inhibition of base line and Ca" - stimulated 86 Rb efflux from T84 monolayers. Detailed Description of the Invention 30 As discussed in the Background section, blockade of sickle dehydration via inhibition of the Gardos channel is a powerful therapeutic approach towards the treatment and/or prevention of sickle cell disease. In vitro studies have shown that Clotrimazole, an imidazole- WO 99/26628 PCT/US98/24967 - 15 containing antimycotic agent, blocks Ca 2 -activated K + transport and cell dehydration in sickle erythrocytes (Brugnara et al., 1993, J. Clin. Invest. 92:520-526). Studies in a transgenic mouse model for sickle cell disease (SAD mouse, Trudel et al., 1991, EMBO J. 11:3157 3165) show that oral administration of Clotrimazole leads to inhibition of the red cell Gardos 5 channel, increased red cell K' content, a decreased mean cell hemoglobin concentration (MCHC) and decreased cell density (De Franceschi et al., 1994, J. Clin. Invest. 93:1670 1676). Moreover, therapy with oral Clotrimazole induces inhibition of the Gardos channel and reduces erythrocyte dehydration in patients with sickle cell disease (Brugnara et al., 1996, J. Clin. Invest. 97:1227-1234). Other antimycotic agents which inhibit the Gardos channel in 10 vitro include miconazole, econazole, butoconazole, oxiconazole and sulconazole (U.S. Patent No. 5,273,992 to Brugnara et al.). All of these compounds contain an imidazole-like ring, i.e., a heteroaryl ring containing two or more nitrogens. Also as discussed in the Background section, the modulation of early ionic mitogenic signals and inhibition of cell proliferation are powerful therapeutic approaches towards the 15 treatment and/or prevention of disorders characterized by abnormal cell proliferation. It has been shown that Clotrimazole, in addition to inhibiting the Gardos channel of erythrocytes, also modulates ionic mitogenic signals and inhibits cell proliferation both in vitro and in vivo. For example, Clotrimazole inhibits the rate of cell proliferation of normal and cancer cell lines in a reversible and dose-dependent manner in vitro (Benzaquen et al., 1995 Nature 20 Medicine 1:534-540). Clotrimazole also depletes the intracellular Ca 2 + stores and prevents the rise in cystolic Ca 2+ that normally follows mitogenic stimulation. Moreover, in mice with severe combined immunodeficiency disease (SCID) and inoculated with MM-RU human melanoma cells, daily administration of Clotrimazole resulted in a significant reduction in the number of lung metastases observed (Benzaquen et al., supra). 25 The discovery that 11-phenyl dibenzazepine compounds, and analogues thereof, inhibit the Gardos channel of erythrocytes, mammalian cell proliferation and/or Cl- secretion from intestinal cells, was quite surprising. Thus, in one aspect, the present invention provides a new class of organic compounds that are capable of inhibiting the Gardos channel of erythrocytes, mammalian cell proliferation, particularly mitogen-induced cell proliferation, 30 and/or Cl- secretion from intestinal cells. Significantly, the compounds of the invention do not contain an imidazole or imidazole-like moiety. The imidazole or imidazole-like moiety is well-recognized as the WO 99/26628 PCT/US98/24967 - 16 essential functionality underlying the antimycotic and other biological activities of Clotrimazole and the other above-mentioned anti-mycotic agents. Thus, the 11-phenyl dibenzazepine compounds of the invention provide an entirely new class of compounds that are capable of effecting inhibition of the Gardos channel of erythrocytes, mammalian cell 5 proliferation, particularly mitogen-induced cell proliferation, and/or Cl- secretion from intestinal cells. In another aspect, the invention provides a method of reducing sickle cell dehydration and/or delaying the occurrence of erythrocyte sickling in situ as a therapeutic approach towards the treatment of sickle cell disease. In its broadest sense, the method involves only a 10 single step -- the administration of at least one pharmacologically active compound of the invention, or a composition thereof, to a sickle erythrocyte in situ in an amount effective to reduce dehydration and/or delay the occurrence of cell sickling or deformation. While not intending to be bound by any particular theory, it is believed that administration of the active compounds described herein in appropriate amounts to sickle 15 erythrocytes in situ causes nearly complete inhibition of the Gardos channel of sickle cells, thereby reducing the dehydration of sickle cells and/or delaying the occurrence of cell sickling or deformation. In a preferred embodiment, the dehydration of a sickle cell is reduced and/or the occurrence of sickling is delayed in a sickle cell that is within the microcirculation vasculature of the subject, thereby reducing or eliminating the vaso-occlusion that is 20 commonly caused by sickled cells. Based in part on the surmised importance of the Gardos channel as a therapeutic target in the treatment of sickle cell disease, the invention is also directed to methods of treating or preventing sickle cell disease. In the method, an effective amount of one or more compounds according to the invention, or a pharmaceutical composition thereof, is administered to a 25 patient suffering from sickle cell disease. The methods may be used to treat sickle cell disease prophylactically to decrease intracellular Hb S concentration and/or polymerization, and thus diminish the time and duration of red cell sickling and vaso-occlusion in the blood circulation. The methods may also be used therapeutically in patients with acute sickle cell crisis, and in patients suffering chronic sickle cell episodes to control both the frequency and duration of 30 the crises. The compounds of the invention are also potent, specific inhibitors of mammalian cell proliferation. Thus, in another aspect, the invention provides methods of inhibiting WO 99/26628 PCT/US98/24967 -17 mammalian cell proliferation as a therapeutic approach towards the treatment or prevention of diseases characterized by unwanted or abnormal cell proliferation. In its broadest sense, the method involves only a single step -- the administration of an effective amount of at least one pharmacologically active compound according to the invention to a mammalian cell in situ. 5 The compound may act cytostatically, cytotoxically, or by a combination of both mechanisms to inhibit cell proliferation. Mammalian cells treatable in this manner include vascular smooth muscle cells, fibroblasts, endothelial cells, various pre-cancer cells and various cancer cells. In a preferred embodiment, cell proliferation is inhibited in a subject suffering from a disorder that is characterized by unwanted or abnormal cell proliferation. Such diseases are 10 described more fully below. Based in part on the surmised role of mammalian cell proliferation in certain diseases, the invention is also directed to methods of treating or preventing diseases characterized by abnormal cell proliferation. In the method, an effective amount of at least one compound according to the invention, or a pharmaceutical composition thereof, is administered to a 15 patient suffering from a disorder that is characterized by abnormal cell proliferation. While not intending to be bound by any particular theory, it is believed that administration of an appropriate amount of a compound according to the invention to a subject inhibits cell proliferation by altering the ionic fluxes associated with early mitogenic signals. Such alteration of ionic fluxes is thought to be due to the ability of the compounds of the invention 20 to inhibit potassium channels of cells, particularly Ca2+-activated potassium channels. The method can be used prophylactically to prevent unwanted or abnormal cell proliferation, or may be used therapeutically to reduce or arrest proliferation of abnormally proliferating cells. The compound, or a pharmaceutical formulation thereof, can be applied locally to proliferating cells to arrest or inhibit proliferation at a desired time, or may be administered to 25 a subject systemically to arrest or inhibit cell proliferation. Diseases which are characterized by abnormal cell proliferation that can be treated or prevented by means of the present invention include blood vessel proliferative disorders, fibrotic disorders, arteriosclerotic disorders and various cancers. Blood vessel proliferation disorders refer to angiogenic and vasculogenic disorders 30 generally resulting in abnormal proliferation of blood vessels. The formation and spreading of blood vessels, or vasculogenesis and angiogenesis, respectively, play important roles in a variety of physiological processes such as embryonic development, corpus luteum formation, WO 99/26628 PCT/US98/24967 - 18 wound healing and organ regeneration. They also play a pivotal role in cancer development. Other examples of blood vessel proliferative disorders include arthritis, where new capillary blood vessels invade the joint and destroy cartilage and ocular diseases such as diabetic retinopathy, where new capillaries in the retina invade the vitreous, bleed and cause blindness 5 and neovascular glaucoma. Another example of abnormal neovascularization is that associated with solid tumors. It is now established that unrestricted growth of tumors is dependent upon angiogenesis and that induction of angiogenesis by liberation of angiogenic factors can be an important step in carcinogenesis. For example, basic fibroblast growth factor (bFGF) is liberated by several 10 cancer cells and plays a crucial role in cancer angiogenesis. The demonstration that certain animal tumors regress when angiogenesis is inhibited has provided the most compelling evidence for the role of angiogenesis in tumor growth. Other cancers that are associated with neovascularization include hemangioendotheliomas, hemangiomas and Kaposi's sarcoma. Proliferation of endothelial and vascular smooth muscle cells is the main feature of 15 neovascularization. The invention is useful in inhibiting such proliferation, and therefore in inhibiting or arresting altogether the progression of the angiogenic condition which depends in whole or in part upon such neovascularization. The invention is particularly useful when the condition has an additional element of endothelial or vascular smooth muscle cell proliferation that is not necessarily associated with neovascularization. For example, psoriasis 20 may additionally involve endothelial cell proliferation that is independent of the endothelial cell proliferation associated with neovascularization. Likewise, a solid tumor which requires neovascularization for continued growth may also be a tumor of endothelial or vascular smooth muscle cells. In this case, growth of the tumor cells themselves, as well as the neovascularization, is inhibited by the compounds described herein. 25 The invention is also useful for the treatment of fibrotic disorders such as fibrosis and other medical complications of fibrosis which result in whole or in part from the proliferation of fibroblasts. Medical conditions involving fibrosis (other than atherosclerosis, discussed below) include undesirable tissue adhesion resulting from surgery or injury. Other cell proliferative disorders which can be treated by means of the invention 30 include arteriosclerotic conditions. Arteriosclerosis is a term used to describe a thickening and hardening of the arterial wall. An arteriosclerotic condition as used herein means classical atherosclerosis, accelerated atherosclerosis, atherosclerotic lesions and any other WO 99/26628 PCT/US98/24967 -19 arteriosclerotic conditions characterized by undesirable endothelial and/or vascular smooth muscle cell proliferation, including vascular complications of diabetes. Proliferation of vascular smooth muscle cells is a main pathological feature in classical atherosclerosis. It is believed that liberation of growth factors from endothelial cells 5 stimulates the proliferation of subintimal smooth muscle which, in turn, reduces the caliber and finally obstructs the artery. The invention is useful in inhibiting such proliferation, and therefore in delaying the onset of, inhibiting the progression of, or even halting the progression of such proliferation and the associated atherosclerotic condition. Proliferation of vascular smooth muscle cells produces accelerated atherosclerosis, 10 which is the main reason for failure of heart transplants that are not rejected. This proliferation is also believed to be mediated by growth factors, and can ultimately result in obstruction of the coronary arteries. The invention is useful in inhibiting such obstruction and reducing the risk of, or even preventing, such failures. Vascular injury can also result in endothelial and vascular smooth muscle cell 15 proliferation. The injury can be caused by any number of traumatic events or interventions, including vascular surgery and balloon angioplasty. Restenosis is the main complication of successful balloon angioplasty of the coronary arteries. It is believed to be caused by the release of growth factors as a result of mechanical injury to the endothelial cells lining the coronary arteries. Thus, by inhibiting unwanted endothelial and smooth muscle cell 20 proliferation, the compounds described herein can be used to delay, or even avoid, the onset of restenosis. Other atherosclerotic conditions which can be treated or prevented by means of the present invention include diseases of the arterial walls that involve proliferation of endothelial and/or vascular smooth muscle cells, such as complications of diabetes, diabetic 25 glomerulosclerosis and diabetic retinopathy. The compounds described herein are also useful in treating or preventing various types of cancers. Cancers which can be treated by means of the present invention include, but are not limited to, biliary tract cancer; brain cancer, including glioblastomas and medulloblastomas; breast cancer; cervical cancer; choriocarcinoma; colon cancer; endometrial 30 cancer; esophageal cancer; gastric cancer; hematological neoplasms, including acute and chronic lymphocytic and myelogenous leukemia, multiple myeloma, AIDS associated leukemias and adult T-cell leukemia lymphoma; intraepithelial neoplasms, including Bowen's WO 99/26628 PCT/US98/24967 - 20 disease and Paget's disease; liver cancer; lung cancer; lymphomas, including Hodgkin's disease and lymphocytic lymphomas; neuroblastomas; oral cancer, including squamous cell carcinoma; ovarian cancer, including those arising from epithelial cells, stromal cells, germ cells and mesenchymal cells; pancreas cancer; prostate cancer; rectal cancer; sarcomas, 5 including leiomyosarcoma, rhabdomyosarcoma, liposarcoma, fibrosarcoma and osteosarcoma; skin cancer, including melanoma, Kaposi's sarcoma, basocellular cancer and squamous cell cancer; testicular cancer, including germinal tumors (seminoma, non-seminoma (teratomas, choriocarcinomas)), stromal tumors and germ cell tumors; thyroid cancer, including thyroid adenocarcinoma and medullar carcinoma; and renal cancer including adenocarcinoma and 10 Wilms tumor. The compounds of the invention are useful with hormone dependent and also with nonhormone dependent cancers. They also are useful with prostate and nonprostate cancers and with breast and nonbreast cancers. They further are useful with multidrug resistant strains of cancer. 15 In addition to the particular disorders enumerated above, the invention is also useful in treating or preventing dermatological diseases including keloids, hypertrophic scars, seborrheic dermatosis, papilloma virus infection (e.g., producing verruca vulgaris, verruca plantaris, verruca plan, condylomata, etc.), and eczema and epithelial precancerous lesions such as actinic keratosis. It also is useful with pathologies mediated by growth factors such as 20 uterine leiomyomas. In addition to the particular disorders enumerated above, the invention is particularly useful in treating or preventing inflammatory diseases associated with cellular proliferation. An "inflammatory disease associated with cellular proliferation" as used herein is a disease in which lymphoproliferation contributes to tissue or organ damage leading to disease. For 25 instance, excessive T cell proliferation at the site of a tissue or organ will cause damage to the tissue or organ. Inflammatory disease are well known in the art and have been described extensively in medical textbooks (See, e.g., Harrison's Principles of Experimental Medicine, 13th Edition, McGraw-Hill, Inc., N.Y.). Inflammatory diseases associated with cellular proliferation include but are not limited 30 to proliferative glomerulonephritis; lupus erythematosus; scleroderma; temporal arteritis; thromboangiitis obliterans; mucocutaneous lymph node syndrome; asthma; host versus graft; inflammatory bowel disease; multiple sclerosis; rheumatoid arthritis; thyroiditis; Grave's WO 99/26628 PCT/US98/24967 -21 disease; antigen-induced airway hyperactivity; pulmonary eosinophilia; Guillain-Barre syndrome; allergic rhinitis; myasthenia gravis; human T-lymphotrophic virus type 1 associated myelopathy; herpes simplex encephalitis; inflammatory myopathies; atherosclerosis; and Goodpasture's syndrome. Some examples of inflammatory diseases 5 associated with cellular proliferation as well as animal models for testing and developing the compounds are set forth in Table 1 below. Table 1 Disease Proliferating Cells Reference Animal Model Reference Asthma T cells Hogg 1997 APMIS Airway inflammation Henderson et al. 100:105(10):735-45 and 1997 J Clin Invest hyperresponsiveness 100(12):3083-3092. in Ovalbumin sensitized mice or guinea pigs. 10 Glomerulonephritis Mesangial Nitta et al. 1998 NZB/NZW crossed Clynes et al. 1998 (glomerular) cells Eur J Pharmacol mice develop Science 279(5353): 344:107-110 glomerular disease 1052-54. and lupus-like syndrome. Host versus Graft T cells Schorlemmer et al. Renal allograft Lazarivuts et al. 1996 B cells 1997 Int J Tissue rejection in mice. Nature 380(6576) React 19:157-61. 717-720. Sedgwick et al. 1998 J Immunol 160:5320-30. Inflammatory Bowel Epithelial cells Bajaj-Elliott et al. Trinitrobenzene Boughton-Smith et Disease 1997 Am J. Pathol. sulphonic acid al. 1988 Br J 151:1469-76. induced bowel Pharmacol 94:65-72. inflammation in rats. Systemic Lupus Glomerular cells Kodera et al. 1997 NZB/NZW crossed Peng et al. 1996 15 Erythematosis Lymphocytes Am J Nephol 17:466- mice develop Mol Biol Rep 23(3 70. glomerular disease 4):247-51. Akashi et al. 1998 and lupus-like Immunology 93:238- syndrome. 48 Multiple Sclerosis T cells Constantinesecu et Experimental allergic Drescher et al. 1998 al. 1998 Immunol encephalomyelitis. J Clin Invest Res 17(1-2):217-27. 101(8):1765-74. Rheumatoid Arthritis T cells Ceponis et al. 1998 Rat adjuvant arthritis Anderson et al. 1996 Synovial cells Br J Rheumatol assay J Clin Invest 37(2):170-8 97(11):2672-9. Thyroiditis T cells and Rose et al. 1997 HLA transgenic mice Taneja et al. 1998 Epithelial cells Crit Rev Immunol immunized with J Clin Investig 17:511-7. thyroglobulin. 101(5):921-6. Schumm-Draeger et al. 1996 Verh Dtsch Ges Pathol 80:297 301.

WO 99/26628 PCT/US98/24967 - 22 Grave's Disease Thyroid cells DiPaola et al. 1997 Thiouracil-fed rats. Viglietto et al. 1997 J Clin Endocrinol Oncogene 15:2687 Metab 82:670-3. 98. Disease Proliferating Cells Reference Model Antigen-induced airway T cells Wolyniec et al. 1998 5 hyperactivity Am J Respir Cell Mol Biol 18:777-85 Pulmonary eosinophilia T cells Wolyniec et al. 1998 Am J Respir Cell Mol Biol 18:777-85 Guillain-Barre T cells Hartung et al. 1991 Experimental Syndrome Ann Neurol. 30:48-53 autoimmune neuritis (inflammatory (immunization with PNS 10 demyelinating disease) myelin and Freunds complete adjuvant) Giant cell arteritis (a T cells Brack et al. 1997 form of systemic Mol Med 3:530-43 vasculitis) Inflammation of large arteries 15 Allergic Rhinitis T cells Baraniuk et al. 1997 J Allergy Clin Immunol 99:S763-72 Myasthenia gravis T cells Hartung et al. 1991 Ann Neurol 30:48-53 Human T-lymphotropic T cells Nakamura et al. 1996 virus type 1 - associated Intern Mede 35:195-99 myelopathy 20 Herpes simplex T cells Hartung et al. 1991 encephalitis Ann Neurol 30:48-53 Inflammatory T cells Hartung et al. 1991 myopathies (ie. Ann Neurol 30:48-53. Polymyositis, Lindberg et al. 1995 25 dermatomyocitis) Scand J Immunol 41:421-26 Artherosclerosis T cells Rosenfeld et al. 1996 Diabetes Res Clin Pract 30 suppl.: 1-11 Goodpasture's syndrome Macrophages Lan et al. 1995 Am J Pathol 147:1214 20 The compounds and methods of the invention provide myriad advantages over agents 30 and methods commonly used to treat cell proliferative disorders. For example, many of the compounds of the invention are more potent than Clotrimazole in in vitro tests, and therefore WO 99/26628 PCT/US98/24967 - 23 may provide consequential therapeutic advantages in clinical settings. Most significantly, the compounds of the invention have reduced toxicity as compared with these other agents. For Clotrimazole, it is well-known that the imidazole moiety is responsible for inhibiting a wide range of cytochrome P-450 isozyme catalyzed reactions, 5 which constitutes their main toxicological effects (Pappas and Franklin, 1993, Toxicology 80:27-35; Matsuura et al., 1991, Biochemical Pharmacology 41:1949-1956). Analogues and metabolites of Clotrimazole do not induce cytochrome P-450 (Matsuura et al., 1991, Biochemical Pharmacology 41:1949-1956), and therefore do not share Clotrimazole's toxicity. The invention in another aspect also involves methods and products for reducing the 10 symptoms of diarrhea or preventing diarrhea in a subject at risk for developing diarrhea, using the compounds of the invention. The aromatic compounds useful according to the invention may be provided in a pharmaceutical preparation or a veterinary preparation. The aromatic compounds of the invention are also useful in a method for treating diarrhea and scours as well as a method for preventing diarrhea and scours. 15 Diarrhea, as used herein, indicates a medical syndrome which is characterized by the symptoms of diarrhea or scours. In general, diarrhea is a disorder resulting in a secretory imbalance. For purposes of this patent application diarrhea is divided into three categories based on the underlying mechanism: exudative, decreased absorption, and secretory and the term diarrhea as used herein encompasses each of these categories. Exudative diarrheas result 20 from inflammatory processes leading to impaired colonic absorption, and outpouring of cells and colloid caused by such disorders as ulcerative colitis, shigellosis, and amebiasis. Disorders of decreased absorption include osmotic, anatomic derangement, and motility disorders. Osmotic diarrhea can occur as a result of digestive abnormalities such as lactose intolerance. Anatomic derangement results in a decreased absorption surface caused by such 25 procedures as subtotal colectomy and gastrocolic fistula. Motility disorders result from decreased contact time resulting from such diseases as hyperthyroidism and irritable bowel syndrome. Secretory diarrhea is characterized by the hypersecretion of fluid and electrolytes from the cells of the intestinal wall. In classical form, the hypersecretion is due to changes which are independent of the permeability, absorptive capacity and exogenously generated 30 osmotic gradients within the intestine. As discussed above, however, all forms of diarrhea may actually manifest a secretory component. The methods and products of the invention are particularly useful in treating diarrhea WO 99/26628 PCT/US98/24967 - 24 which is secretory. However, the methods and products of the invention may also be used in combination with other treatment methods which are known in the art to treat diarrhea caused by decreased absorption or inflammation. The compounds of the invention are involved in regulating Cl- secretion and can function alone or when used in combination with other 5 treatment methods to decrease net fluid secretion even when this is due primarily to abnormalities in absorption or inflammation. The methods and products of the invention are useful in preventing diarrhea and scours in subjects at risk of developing these disorders. Subjects at risk of developing diarrhea and scours are those subjects which have a high likelihood of exposure to the 10 bacterial and viral microorganisms which cause these symptoms. For example, approximately 1/3 of travelers to developing countries will develop diarrhea; infection with rotavirus is one of the leading causes of death in infants in developing countries; subjects with HIV have a greater than 50% chance of developing diarrhea, and many newborn calves and pigs develop scours. Subjects with inflammatory bowel disease develop recurrent diarrhea. 15 The methods and products of the invention are also useful in treating subjects who already exhibit the symptoms of diarrhea and scours. Once a subject has been exposed to a microorganism causing the symptoms, the subject may be treated with the methods and products of the present invention in order to reduce the symptoms. The symptoms of diarrhea include bowel irregularity, fecal fluid rich in sodium or potassium, fluid feces, dehydration, 20 fever, loss of body weight, headache, anorexia, vomiting, malaise and myalgia. The symptoms of scours include a loss of body weight or failure to grow, dehydration, malodorous feces, fluid feces, feces containing pieces of partially digested milk or semisolid material, and feces of a yellow-white or gray color. The Compounds 25 The compounds which are potent, selective and safe inhibitors of Ca2+-activated potassium channel (Gardos channel) of erythrocytes, particularly sickle erythrocytes, mammalian cell proliferation, particularly mitogen-induced cell proliferation, and!or secretagogue-stimulated transepithelial electrogenic chloride secretion in intestinal cells according to the invention, are generally substituted 11-phenyl dibenzazepine compounds. 30 In one illustrative embodiment, the compounds capable of inhibiting the Gardos channel, mammalian cell proliferation and/or chloride secretion in intestinal cells according to the invention are compounds having the structural formula (I): WO 99/26628 PCT/US98/24967 - 25 Rq R6 R15 R5 N

R

7 R4 (I) R 9 R R, R2 R3 5 RIo R 14 (I)R R11 R13 R12 10 wherein:

R

1 is -R', (C 6

-C

20 ) aryl or substituted (C 6

-C

2 0 ) aryl;

R

2 is -R', -OR', -SR', halogen or trihalomethyl;

R

3 is -R', -OR', -SR', halogen or trihalomethyl or, when taken together with R 4 , is (C 6 C 2 0 ) aryleno; 15 R 4 is -R', -OR', -SR', halogen or trihalomethyl or, when taken together with R 3 , is (C 6 C 20 ) aryleno; each of Rs, R 6 , R 7 , R 8 , R 9 , Rio, R 11 , R 1 2 , R 1 3 and R 1 4 is independently selected from the group consisting of -R', halogen and trihalomethyl;

R,

5 is -R", -C(O)R", -C(S)R", -C(O)OR", -C(S)OR", -C(O)SR", -C(S)SR", 20 -C(O)N(R") 2 , -C(S)N(R") 2 , -C(O)C(O)R", -C(S)C(O)R", -C(O)C(S)R", -C(S)C(S)R", -C(O)C(O)OR", -C(S)C(O)OR", -C(O)C(S)OR", -C(O)C(O)SR", -C(S)C(S)OR", -C(S)C(O)SR", -C(O)C(S)SR", -C(S)C(S)SR", -C(O)C(O)N(R") 2 , -C(S)C(O)N(R") 2 ,

-C(O)C(S)N(R")

2 or -C(S)C(S)N(R")2; each R' is independently selected from the group consisting of-H, (C 1

-C

6 ) alkyl, 25 (C 1

-C

6 ) alkenyl and (C 1

-C

6 ) alkynyl; each R" is independently selected from the group consisting of -H, (Ci-C 6 ) alkyl,

(C

1

-C

6 ) alkenyl, (C 1

-C

6 ) alkynyl, (C 6

-C

20 ) aryl, (C 6

-C

2 0 ) substituted aryl, (C 6

-C

26 ) alkaryl and substituted (C 6

-C

26 ) alkaryl; and the aryl and alkaryl substituents are each independently selected from the group 30 consisting of-CN, -OR', -SR', -NO 2 , -NR'R', halogen, (C 1

-C

6 ) alkyl, (C 1

-C

6 ) alkenyl, (C 1

-C

6 ) alkynyl and trihalomethyl. In a preferred embodiment of the invention, the compounds are those of structure (I) WO 99/26628 PCT/US98/24967 - 26 wherein the chalcogens are each oxygen. In another preferred embodiment, the compounds are those of structure (I) wherein the halogens are each independently -F, -Cl, -Br or -I. In another preferred embodiment, the alkyl, alkenyl and alkynyl groups are each 5 independently (CI-C 3 ) and/or the aryl groups are phenyl and/or the aryleno groups are benzeno. In another preferred embodiment, Rs, R 6 , R 7 , R 9 , Rio, RI, and R, 3 are each independently -R'. In another preferred embodiment, the substituted aryl and alkaryl are mono 10 substituted. In another preferred embodiment, R 1 5 is -R", -C(O)R", -C(O)OR", -C(O)N(R") 2 , -C(O)C(O)R", -C(O)C(O)OR" or -C(O)C(O)N(R") 2 . In another preferred embodiment of the invention, the compounds are those of structural formula (I) wherein: 15 R 1 is -R' or (C 6

-C

20 ) aryl;

R

2 is -R' or -OR';

R

3 is -R' or -OR' or, when taken together with R 4 , is (C 6

-C

20 ) aryleno;

R

4 is -R' or -OR' or, when taken together with R 3 , is (C 6

-C

20 ) aryleno; each of Rs, R 6 , R 7 , R 8 , R 9 , RIO, Ri 1 , R 1 2 , R 1 3 and R 14 is independently selected from the 20 group consisting of -R' and halogen;

R

,5 is -R", -C(O)R", -C(O)OR", -C(O)N(R") 2 , -C(O)C(O)R", -C(O)C(O)OR" or -C(O)C(O)N(R")2; each R' is independently selected from the group consisting of -H, (Ci-C 6 ) alkyl,

(C

1

-C

6 ) alkenyl and (C 1

-C

6 ) alkynyl; 25 each R" is independently selected from the group consisting of -H, (C

I

-C

6 ) alkyl,

(C

1

-C

6 ) alkenyl, (C 1

-C

6 ) alkynyl, (C 6

-C

20 ) aryl, substituted (C 6

-C

20 ) aryl, (C 6

-C

26 ) alkaryl and substituted (C 6

-C

2 6 ) alkaryl; and the aryl and alkaryl substituents are each independently selected from the group consisting of -CN, -OR', -NO 2 , -NR'R', halogen, (C 1

-C

6 ) alkyl, (C,-C 6 ) alkenyl and (C 1

-C

6 ) 30 alkynyl. In another preferred embodiment, the compounds are those of formula (I) wherein:

R

1 is -R' or (C 6

-C

1 0 ) aryl; WO 99/26628 PCT/US98/24967 - 27 R 2 is -R' or -OR';

R

3 is -R' or -OR' or, when taken together with RI 4 , is (C 6

-C

10 ) aryleno;

R

4 is -R' or -OR' or, when taken together with R 3 , is (C 6

-C

1 0 ) aryleno; each of Rs, R 6 and R 7 is -H; 5 R 8 is -R', -F, -Cl, -Br or -I; each of R 9 , Rio and RI is -H;

R

12 is -R', -F, -Cl, -Br or -I;

R

13 is -H;

RI

4 is -R', -F, -Cl, -Br or -I; 10 Rs is -R", -C(O)R", -C(O)OR", -C(O)N(R") 2 , -C(O)C(O)R", -C(O)C(O)OR" or -C(O)C(O)N(R")2; each R' is independently selected from the group consisting of-H, (C 1

-C

3 ) alkyl,

(C

1

-C

3 ) alkenyl and (CI-C 3 ) alkynyl; each R" is independently selected from the group consisting of-H, (Ci-C 3 ) alkyl, 15 (C 1

-C

3 ) alkenyl, (C 1

-C

3 ) alkynyl, (C 6 -Co 10 ) aryl, substituted (C 6

-C

10 ) aryl, (C 6

-C

1 3 ) alkaryl or substituted (C 6

-C

1 3 ) alkaryl; and the aryl and alkaryl substituents are each independently selected from the group consisting of-OR',-NO 2 , -NR'R', -F, -Cl, -Br, -I, (C 1

-C

3 ) alkyl, (C 1

-C

3 ) alkenyl and (C 1

-C

3 ) alkynyl. 20 In still another preferred embodiment, the compounds are those of structural formula (I) wherein: R, is -R' or phenyl;

R

2 is -R' or -OR';

R

3 is -R' or -OR' or, when taken together with R 4 , is benzeno; 25 R 4 is -R' or -OR' or, when taken together with R 3 , is benzeno; each of Rs, R 6 and R 7 is -H;

R

8 is -R', -Cl or -Br; each of R 9 , Rio and R, is -H;

R

1 2 is -R', -F or -Cl; 30 R 13 is -H;

R

14 is -R'or -Cl;

R

15 is -R", -C(O)R", -C(O)OR", -C(O)NHR", -C(O)C(O)R" or -C(O)C(O)OR"; WO 99/26628 PCT/US98/24967 -28 each R' is independently selected from the group consisting of -H, (C

,

-C

3 ) alkyl,

(C

1

-C

3 ) alkenyl and (C 1

-C

3 ) alkynyl; each R" is independently selected from the group consisting of -H, (C I-C 3 ) alkyl,

(C

1

-C

3 ) alkenyl, (C 1

-C

3 ) alkynyl, (C 6

-C

0 ) aryl, mono-substituted (C 6

-CI

0 ) aryl, (C 6

-C

1 3 ) alkaryl 5 or mono-substituted (C 6

-C

1 3 ) alkaryl; and the aryl and alkaryl substituents are each independently selected from the group consisting of-OR', -NO 2 , -NR'R', -Cl, (C

,

-C

3 ) alkyl, (CI-C 3 ) alkenyl and (Ci-C 3 )alkynyl. As used herein, the term "alkyl" refers to a saturated branched, straight chain or cyclic hydrocarbon radical. Typical alkyl groups include methyl, ethyl, propyl, isopropyl, 10 cyclopropyl, butyl, isobutyl, t-butyl, cyclobutyl, pentyl, isopentyl, cyclopentyl, hexyl, cyclohexyl and the like. As used herein, the term "heterocycloalkyl" refers to a saturated cyclic hydrocarbon radical wherein one or more of the carbon atoms is replaced with another atom such as Si, Ge, N, O, S or P. Typical heterocycloalkyl groups include, but are not limited to, morpholino, 15 thiolino, piperidyl, pyrrolidinyl, piperazyl, pyrazolidyl, imidazolidinyl, and the like. As used herein, the term "alkenyl" refers to an unsaturated branched, straight chain or cyclic hydrocarbon radical having at least one carbon-carbon double bond. The radical may be in either the cis or trans conformation about the double bond(s). Typical alkenyl groups include ethenyl, propenyl, isopropenyl, cyclopropenyl, butenyl, isobutenyl, cyclobutenyl, tert 20 butenyl, pentenyl, hexenyl and the like. As used herein, the term "alkynyl" refers to an unsaturated branched, straight chain or cyclic hydrocarbon radical having at least one carbon-carbon triple bond. Typical alkynyl groups include ethynyl, propynyl, butynyl, isobutynyl, pentynyl, hexynyl and the like. As used herein, the term "alkoxy:" refers to an -OR radical, where R is alkyl, alkenyl 25 or alkynyl, as defined above. As used herein, the term "aryl" refers to an unsaturated cyclic hydrocarbon radical having a conjugated n electron system. Typical aryl groups include, but are not limited to, penta-2,4-diene, phenyl, naphthyl, anthracyl, azulenyl, indacenyl, and the like. As used herein, the term "heteroaryl" refers to an aryl group wherein one or more of 30 the ring carbon atoms is replaced with another atom such as N, O or S. Typical heteroaryl groups include, but are not limited to,furanyl, imidazole, pyridinyl, thiophenyl, indolyl, imidazolyl, quinolyl, thienyl, indolyl, pyrrolyl, pyranyl, pyridyl, pyrimidyl, pyrazyl, WO 99/26628 PCT/US98/24967 - 29 pyridazyl, and the like. As used herein, the term "heteroarylium" refers to a heteroaryl group wherein one or more hydrogens has been added to any position of the neutral parent ring. Typical heteroarylium groups include, but are not limited to, pyridinium, pyrazinium, pyrimidinium, 5 pyridazinium, 1,3,5-triazinium, and the like. As used herein, the term "in situ" refers to and includes the terms "in vivo," "ex vivo," and "in vitro" as these terms are commonly recognized and understood by persons ordinarily skilled in the art. Moreover, the phrase "in situ" is employed herein in its broadest connotative and denotative contexts to identify an entity, cell or tissue as found or in place, 10 without regard to its source or origin, its condition or status or its duration or longevity at that location or position. In still another preferred embodiment, the compounds of the invention are as follows: NH HzC (1) (2) N H HzC

HC

N /\ i) N Cl c 15 (3C -\ ( 15 (3) (4) (5) WO 99/26628 PCT/US98/24967 -30 O OCH 3

H

3 C\ NN Br cl (6) (7) (8)

OCH

3 0CH 2

CH

3 HO oKo 0 N O OO oN Cl (9) (10) (11) CI NH O o / \K OCz N ON N F (12) (13) (14)

OCH

3 NH /\ /\\ o 0 N N cl\ (15) (16) (17) WO 99/26628 PCT/US98/24967 -31 NO 2 NH NH \ / / \ / \ ozN \ / ' CH 0 N CI CI O. CH, cl (18) (19) (20)

OCH

3

CH

3 O 0= O N N N CH3 CI c (21) (22) (23) CI O 0O 0= o-- 0=<O N N N

CH

3 I I (24) (25) (26)

CH

3

NO

2

CH

3 N o 0 N N CI C(27) (28) (29)CI (2'7) (28) (29) WO 99/26628 PCT/US98/24967 - 32 O No 2 O NO 2 NH-O N N 332 o CH CH C 1Cl C O. 0 1 CH H 3 C1 (30) (31) (32)

NH-CH

3 NHN O NH

NO

2 O o N N N/\ X C1 C1 C1 (33) (34) (35) The compounds will be referred to herein by way of compound numbers as presented 5 above. In yet another preferred embodiment, the compounds are those of structural formula (I), with the proviso that when R, and R 1 5 are each -R', at least one of R 1 , R 2 , R 3 , R 4 , Rs, R 6 ,

R

7 , R 9 , Ro 10 , Ru, R 1 2 , R 1 3 or R 14 is other than -R', R 8 is other than -R' or halogen and at least three of R 2 , R 3 , R 4 and R 5 are other than -OR'. 10 In yet another preferred embodiment, the compounds are those of structural formula (I), with the proviso that when R, and R 1 5 are each -H, at least one of R 1 , R 2 , R 3 , R 4 , Rs, R 6 , R 7 ,

R

9 , RI 0 , R 1 1 , RI 2 , RB 3 or R 14 is other than -H, R 8 is other than -H or -Cl and at least three of R 2 ,

R

3 , R 4 and R 5 are other than -OCH 3 . In a final preferred embodiment, the compounds of the invention are not 11-phenyl 15 5,6-dihydro-11H-dibenz[b,e]azepine, 11-phenyl-9-halo-5,6-dihydro-11H-dibenz[b,e]azepine, 11-phenyl-9-chloro-5,6-dihydro-11H-dibenz[b,e]azepine, 11-phenyl-5,6-dihydro-1,2,3 trialkoxy- 11H-dibenz[b,e]azepine, 11-phenyl-5,6-dihydro-1,2,3-trimethoxy- 11H dibenz[b,e]azepine, 11-phenyl-5,6-dihydro-2,3,4-trialkoxy- 11H-dibenz[b,e]azepine and/or 11 phenyl-5,6-dihydro-2,3,4-trimethoxy- 11 H-dibenz[b,e]azepine. 20 The chemical formulae referred to herein may exhibit the phenomena of tautomerism WO 99/26628 PCT/US98/24967 -33 or conformational isomerism. As the formulae drawings within this specification can represent only one of the possible tautomeric or conformational isomeric forms, it should be understood that the invention encompasses any tautomeric or conformational isomeric forms which exhibit biological or pharmacological activity as described herein. 5 The compounds of the invention may be in the form of free acids, free bases or pharmaceutically effective salts thereof. Such salts can be readily prepared by treating a compound with an appropriate acid. Such acids include, by way of example and not limitation, inorganic acids such as hydrohalic acids (hydrochloric, hydrobromic, etc.), sulfuric acid, nitric acid, phosphoric acid, etc.; and organic acids such as acetic acid, propanoic acid, 10 2-hydroxyacetic acid, 2-hydroxypropanoic acid, 2-oxopropanoic acid, propandioic acid, butandioic acid, etc. Conversely, the salt can be converted into the free base form by treatment with alkali. In addition to the above-described compounds and their pharmaceutically acceptable salts, the invention may employ, where applicable, solvated as well as unsolvated forms of the 15 compounds (e.g. hydrated forms). The compounds described herein may be prepared by any processes known to be applicable to the preparation of chemical compounds. Suitable processes are well known in the art. Preferred processes are illustrated by the representative examples. Additional methods are described in copending application Serial Number unknown, entitled 20 "SYNTHESIS OF 11-ARYL-5,6-DIHYDRO-11H[b,e]AZEPINES, filed Nov. 20, 1997, and US application serial no. 08/975,594, filed November 20, 1997, entitled "Method for the Treatment or Prevention of Sickle Cell Disease with Substituted Phenyl-Dibenzazepine Compounds" which are both incorporated herein by reference in its entirety. Necessary starting materials may be obtained commercially or by standard procedures of organic 25 chemistry. Moreover, many of the compounds are commercially available. An individual compound's relevant activity and potency as an agent to affect sickle cell dehydration or deformation, mammalian cell proliferation and/or secretagogue-stimulated transepithelial electrogenic chloride secretion in intestinal cells may be determined using standard techniques. Preferentially, a compound is subject to a series of screens to determine 30 its pharmacological activity. In most cases, the active compounds of the invention exhibit three pharmacological activities: inhibition of the Gardos channel of erythrocytes, inhibition of secretagogue- WO 99/26628 PCT/US98/24967 - 34 stimulated transepithelial electrogenic chloride secretion in intestinal cells and inhibition of mammalian cell proliferation. However, in some cases, the compounds of the invention may exhibit only one of these pharmacological activities. Any compound encompassed by formula (I) which exhibits at least one of these pharmacological activities is considered to be within 5 the scope of the present invention. In general, the active compounds of some embodiments of the invention are those which induce at least about 25% inhibition of the Gardos channel of erythrocytes (measured at about 10 [M), at least about 25% inhibition of secretagogue-stimulated transepithelial electrogenic chloride secretion in intestinal cells (measured at about 10 [M) and/or about 25% 10 inhibition of mammalian cell proliferation (measured at about 10 [M), as measured using in vitro assays that are commonly known in the art (see, e.g., Brugnara et al., 1993, J. Biol. Chem. 268(12):8760-8768; Benzaquen et al., 1995, Nature Medicine 1:534-540). Alternatively, or in addition, the active compounds of the invention generally will have an

IC

5 0 (concentration of compound that yields 50% inhibition) for inhibition of the Gardos 15 channel of erythrocytes of less than about 10 pM, an IC 5 0 for secretagogue-stimulated transepithelial electrogenic chloride secretion in intestinal cells of less than about 10 pM, and/or an IC 5 0 for inhibition of cell proliferation of less than about 10 pM, as measured using in vitro assays that are commonly known in the art (see, e.g., Brugnara et al., 1993, J. Biol. Chem. 268(12):8760-8768; Benzaquen et al., 1995, Nature Medicine 1:534-540) and the 20 Examples section below. Other assays for assessing the activity and/or potency of an agent with respect to the uses of the invention are described below with respect to an effective amount of the compounds. Representative active compounds according to the invention include Compounds 1 through 35, as illustrated above. 25 In certain embodiments of the invention, compounds which exhibit only one pharmacological activity, or a higher degree of one activity, may be preferred. Thus, when the compound is to be used in methods to treat or prevent sickle cell disease, or in methods to reduce sickle cell dehydration and/or delay the occurrence of erythrocyte sickling or deformation in situ, it is preferred that the compound exhibit at least about 75% Gardos 30 channel inhibition (measured at about 10 [M) and/or have an IC 50 of Gardos channel inhibition of less than about 1 pM, with at least about 90% inhibition and/or an IC 5 0 of less than about 0.1 jM being particularly preferred.

WO 99/26628 PCT/US98/24967 -35 Exemplary preferred compounds for use in methods related to Gardos channel inhibition and sickle cell disease include Compounds 1, 2, 3, 4, 6, 9, 18, 29 and 35, with Compounds 2, 3, 4, 6, 9, 29 and 35 being particularly preferred. When the compound is to be used in methods to treat or prevent diarrhea and/or 5 scours, it is preferred that the compound exhibit at least about 75% inhibition of Cl- secretion from intestinal cells (measured at about 10 pM) and/or have an ICs 0 of inhibition of C1 secretion from intestinal cells of less than about 1 tM, with at least about 90% inhibition and/or an IC 5 0 of less than about 0.1 pM being particularly preferred. When the compound is to be used in methods to treat or prevent disorders 10 characterized by abnormal cell proliferation or in methods to inhibit cell proliferation in situ, it is preferable that the compound exhibit at least about 75% inhibition of mitogen-induced cell proliferation (measured at about 10 tM) and/or have an IC 50 of cell proliferation of less than about 3 IM, with at least about 90% inhibition and/or an IC 5 0 of less than about 1 pM being particularly preferred. Even more preferred compounds meet both the % inhibition and 15 ICs 50 criteria. Exemplary preferred compounds for use in methods of inhibiting mammalian cell proliferation or for the treatment or prevention of diseases characterized by abnormal cell proliferation include Compounds 2, 3, 4, 7, 8, 9, 13, 14, 16, 17, 18, 19, 20, 21, 22, 26, 27, 28, 29, 30, 31, 32, 33 and 34, with Compounds 14, 26, 28, 29, 30 and 31 being particularly 20 preferred. Formulation and Routes of Administration The compounds described herein, or pharmaceutically acceptable addition salts or hydrates thereof, can be delivered to a patient using a wide variety of routes or modes of administration. Suitable routes of administration include, but are not limited to, inhalation, 25 transdermal, oral, rectal, transmucosal, intestinal and parenteral administration, including intramuscular, subcutaneous and intravenous injections. The compounds described herein, or pharmaceutically acceptable salts and/or hydrates thereof, may be administered singly, in combination with other compounds of the invention, and/or in cocktails combined with other therapeutic agents. Of course, the choice of 30 therapeutic agents that can be co-administered with the compounds of the invention will depend, in part, on the condition being treated. A subject as used herein, means humans, primates, horses, cows, sheep, pigs, goats, WO 99/26628 PCT/US98/24967 -36 cats and dogs. For example, when administered to subjects undergoing cancer treatment, the compounds may be administered in cocktails containing other anti-cancer agents and/or supplementary potentiating agents. The compounds may also be administered in cocktails 5 containing agents that treat the side-effects of radiation therapy, such as anti-emetics, radiation protectants, etc. Anti-cancer drugs that can be co-administered with the compounds of the invention include, e.g., Aminoglutethimide; Asparaginase; Bleomycin; Busulfan; Carboplatin; Carmustine (BCNU); Chlorambucil; Cisplatin (cis-DDP); Cyclophosphamide; Cytarabine 10 HCl; Dacarbazine; Dactinomycin; Daunorubicin HCl; Doxorubicin HCl; Estramustine phosphate sodium; Etoposide (VP-16); Floxuridine; Fluorouracil (5-FU); Flutamide; Hydroxyurea (hydroxycarbamide); Ifosfamide; Interferon Alfa-2a, Alfa 2b, Lueprolide acetate (LHRH-releasing factor analogue); Lomustine (CCNU); Mechlorethamine HCI (nitrogen mustard); Melphalan; Mercaptopurine; Mesna; Methotrexate (MTX); Mitomycin; Mitotane 15 (o.p'-DDD); Mitoxantrone HCl; Octreotide; Plicamycin; Procarbazine HCl; Streptozocin; Tamoxifen citrate; Thioguanine; Thiotepa; Vinblastine sulfate; Vincristine sulfate; Amsacrine (m-AMSA); Azacitidine; Hexamethylmelamine (HMM); Interleukin 2; Mitoguazone (methyl GAG; methyl glyoxal bis-guanylhydrazone; MGBG); Pentostatin; Semustine (methyl CCNU); Teniposide (VM-26); paclitaxel and other taxanes; and Vindesine sulfate. 20 Supplementary potentiating agents that can be co-administered with the compounds of the invention include, e.g., Tricyclic anti-depressant drugs (e.g., imipramine, desipramine, amitriptyline, clomipramine, trimipramine, doxepin, nortriptyline, protriptyline, amoxapine and maprotiline); non-tricyclic and anti-depressant drugs (e.g., sertraline, trazodone and citalopram); Ca" antagonists (e.g., verapamil, nifedipine, nitrendipine and caroverine); 25 Amphotericin (e.g., Tween 80 and perhexiline maleate); Triparanol analogues (e.g., tamoxifen); antiarrhythmic drugs (e.g., quinidine); antihypertensive drugs (e.g., reserpine); Thiol depleters (e.g., buthionine and sulfoximine); and calcium leucovorin. The active compound(s) may be administered per se or in the form of a pharmaceutical composition wherein the active compound(s) is in admixture with one or more 30 pharmaceutically acceptable carriers, excipients or diluents. Pharmaceutical compositions for use in accordance with the present invention may be formulated in conventional manner using one or more physiologically acceptable carriers comprising excipients and auxiliaries which WO 99/26628 PCT/US98/24967 - 37 facilitate processing of the active compounds into preparations which can be used pharmaceutically. Proper formulation is dependent upon the route of administration chosen. For injection, the agents of the invention may be formulated in aqueous solutions, preferably in physiologically compatible buffers such as Hanks's solution, Ringer's solution, 5 or physiological saline buffer. For transmucosal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art. For oral administration, the compounds can be formulated readily by combining the active compound(s) with pharmaceutically acceptable carriers well known in the art. Such 10 carriers enable the compounds of the invention to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions and the like, for oral ingestion by a subject to be treated. Pharmaceutical preparations for oral use can be obtained solid excipient, optionally grinding a resulting mixture, and processing the mixture of granules, after adding suitable auxiliaries, if desired, to obtain tablets or dragee cores. Suitable excipients are, in 15 particular, fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol; cellulose preparations such as, for example, maize starch, wheat starch, rice starch, potato starch, gelatin, gum tragacanth, methyl cellulose, hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, and/or polyvinylpyrrolidone (PVP). If desired, disintegrating agents may be added, such as the cross-linked polyvinyl pyrrolidone, agar, or alginic acid or a salt 20 thereof such as sodium alginate. Dragee cores are provided with suitable coatings. For this purpose, concentrated sugar solutions may be used, which may optionally contain gum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethylene glycol, and/or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures. Dyestuffs or pigments may be added to the tablets or 25 dragee coatings for identification or to characterize different combinations of active compound doses. Pharmaceutical preparations which can be used orally include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol. The push-fit capsules can contain the active ingredients in admixture with filler such 30 as lactose, binders such as starches, and/or lubricants such as talc or magnesium stearate and, optionally, stabilizers. In soft capsules, the active compounds may be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols. In WO 99/26628 PCT/US98/24967 -38 addition, stabilizers may be added. All formulations for oral administration should be in dosages suitable for such administration. For buccal administration,the compositions may take the form of tablets or lozenges formulated in conventional manner. 5 For administration by inhalation, the compounds for use according to the present invention are conveniently delivered in the form of an aerosol spray presentation from pressurized packs or a nebulizer, with the use of a suitable propellant, e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In the case of a pressurized aerosol the dosage unit may be determined 10 by providing a valve to deliver a metered amount. Capsules and cartridges of e.g. gelatin for use in an inhaler or insufflator may be formulated containing a powder mix of the compound and a suitable powder base such as lactose or starch. The compounds may be formulated for parenteral administration by injection, e.g., by bolus injection or continuous infusion. Formulations for injection may be presented in unit 15 dosage form, e.g., in ampoules or in multi-dose containers, with an added preservative. The compositions may take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing and/or dispersing agents. Pharmaceutical formulations for parenteral administration include aqueous solutions 20 of the active compounds in water-soluble form. Additionally, suspensions of the active compounds may be prepared as appropriate oily injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, or synthetic fatty acid esters, such as ethyl oleate or triglycerides, or liposomes. Aqueous injection suspensions may contain substances which increase the viscosity of the suspension, such as sodium carboxymethyl 25 cellulose, sorbitol, or dextran. Optionally, the suspension may also contain suitable stabilizers or agents which increase the solubility of the compounds to allow for the preparation of highly concentrated solutions. Alternatively, the active ingredient may be in powder form for constitution with a suitable vehicle, e.g., sterile pyrogen-free water, before use. 30 The compounds may also be formulated in rectal compositions such as suppositories or retention enemas, e.g., containing conventional suppository bases such as cocoa butter or other glycerides.

WO 99/26628 PCT/US98/24967 -39 In addition to the formulations described previously, the compounds may also be formulated as a depot preparation. Such long acting formulations may be administered by implantation or transcutaneous delivery (for example subcutaneously or intramuscularly), intramuscular injection or a transdermal patch. Thus, for example, the compounds may be 5 formulated with suitable polymeric or hydrophobic materials (for example as an emulsion in an acceptable oil) or ion exchange resins, or as sparingly soluble derivatives, for example, as a sparingly soluble salt. The pharmaceutical compositions also may comprise suitable solid or gel phase carriers or excipients. Examples of such carriers or excipients include but are not limited to 10 calcium carbonate, calcium phosphate, various sugars, starches, cellulose derivatives, gelatin, and polymers such as polyethylene glycols. One product of the invention is a veterinary preparation of an aromatic compound of the invention, used alone or combined with an anti-scours agent. An anti-scours agent is a composition which is known to be useful in preventing or inhibiting the symptoms of scours. 15 Known compositions include, for example, colostral extracts, such as those described in U.S. patent no. 4,377,569 and Canadian patent no. 1,175,352 and widely commercially available (e.g. Soluble Colostrum Powder, by VedCo, Inc., St. Joseph MO; Colostrum Bolus II, by RX Veterinary Products, Kansas City MO, etc.); an immunological preparation of colostrum isolated from milk-producing mammals which may have been immunized against certain 20 diarrheal causing microorganisms, such as those described in U.S. patent no. 4,834,974, Australian patent no. 39340/89, Australian patent no. 52547/90, and German patent no. 1,560,344; microorganism specific immunological preparations, including microorganism specific hybridoma-derived monoclonal antibodies such as those described in Sherman et al., Infection and Immunity, V. 42 (2), P. 653-658 (1983) and a bovine immunoglobulin fraction 25 prepared from bovine plasma or clear bovine serum such as the fraction described in U.S. patent no. 3,984,539; oral rehydration fluids and/or replacement electrolyte compositions which are widely commercially available in the form of dry compositions or liquid solutions prepared for oral or intravenous administration (e.g. Electrolyte H, by Agri-Pet Inc., Aubrey TX; Electrolyte Powder 8x, by Phoenix Pharmaceutical Inc, St. Joseph MO; Electrolyte 30 Solution Rx, by Lextron Inc., Greeley CO, ProLabs LTD, St. Joseph MO, and VetTek Inc., Blue Springs MO; Calf Rehydrate, by Durvet Inc., Blue Springs MO, etc.) and antibiotic compositions which are commercially available (e.g. BIOSOL® Liquid, by The UpJohn WO 99/26628 PCT/US98/24967 - 40 Company Animal Health Division, Kalamazoo MI; AMOXI-BOL®, by SmithKline-Beecham Animal Health, Exton PA; 5-WAY CALF SCOUR BOLUS T M , by Agri Laboratories LTD, St. Joseph MO; 1-A-DAY CALF SCOUR BOLUS, by A.H.A.; GARACIN® PIG PUMP, by Schering-Plough Animal Health Corporation, Kenilworth NJ, etc.). 5 In one embodiment, the veterinary preparation is a dry preparation of the aromatic compound of the invention and an antiscours agent. The dry preparation may be administered directly or may be hydrated and/or diluted in a liquid solution prior to administration. In another embodiment, the veterinary preparation is a liquid solution of the compound of the invention and an anti-scours agent. 10 Another product of the invention is a pharmaceutical preparation of an aromatic compound of the invention and an anti-diarrheal agent. An anti-diarrheal agent includes, for example, an immunoglobulin preparation from bovine colostrum; lomotil; an intravenous or oral rehydration fluid; a dry rehydration composition salt; an electrolyte replacement composition (in dry or liquid form); an oral or intravenous sugar-electrolyte solution or dry 15 composition; an antibiotic such as tetracycline, trirmethoprim or sulfamethoxazole; a quinolone drug such as norfloxacin or ciprofloxacin, bismuth subsalicylate, diphenoxylate; and loperamide. In one embodiment the pharmaceutical preparation is a dry preparation of the aromatic compound of the invention and an anti-diarrheal agent. The dry preparation may be 20 administered directly or may be hydrated and/or diluted in a liquid solution prior to administration. In another embodiment the pharmaceutical preparation is a liquid solution of the aromatic compound of the invention and an anti-diarrheal agent. The time of administration of the aromatic compounds useful according to the invention varies depending upon the purpose of the administration. When the compounds of 25 the invention are administered in order to prevent the development of diarrhea in subjects traveling to areas with high risk of exposure to infectious agent or subjects otherwise exposed to diarrhea causing agents, the compounds should be administered at about the time that the subject is exposed to the risk or the high risk area. When the compounds are administered to subjects in order to prevent the development of scours, the veterinary preparation should be 30 administered within the first 12 hours after birth, and preferably within the first 4 hours after birth. When the compounds of the invention are used to treat subjects having symptoms of diarrhea or scours, the compounds may be administered at any point while the subject is WO 99/26628 PCT/US98/24967 -41 experiencing symptoms, and preferably as soon as the symptoms develop. Other considerations will be apparent when the compounds are used to treat inflammatory diseases, proliferative diseases, etc. When administered, the formulations of the invention are applied in pharmaceutically 5 acceptable amounts and in pharmaceutically acceptable compositions. Such preparations may routinely contain salts, buffering agents, preservatives, compatible carriers, and optionally other therapeutic ingredients. When used in medicine the salts should be pharmaceutically acceptable, but non-pharmaceutically acceptable salts may conveniently be used to prepare pharmaceutically acceptable salts thereof and are not excluded from the scope of the 10 invention. Such pharmacologically and pharmaceutically acceptable salts include, but are not limited to, those prepared from the following acids: hydrochloric, hydrobromic, sulfuric, nitric, phosphoric, maleic, acetic, salicylic, p-toluene sulfonic, tartaric, citric, methane sulfonic, formic, malonic, succinic, naphthalene-2-sulfonic, and benzene sulfonic. Also, pharmaceutically acceptable salts can be prepared as alkaline metal or alkaline earth salts, 15 such as sodium, potassium or calcium salts of the carboxylic acid group. Suitable buffering agents include: acetic acid and a salt (1-2% W/V); citric acid and a salt (1-3% W/V); boric acid and a salt (0.5-2.5% W/V); and phosphoric acid and a salt (0.8-2% W/V). Suitable preservatives include benzalkonium chloride (0.003-0.03% W/V); 20 chlorobutanol (0.3-0.9% W/V); parabens (0.01-0.25% W/V) and thimerosal (0.004-0.02% WN). The active compounds of the present invention may be pharmaceutical compositions having a therapeutically effective amount of an aromatic compound of the general formula provided above in combination with a non-formula (I) active agent, optionally included in a 25 pharmaceutically-acceptable carrier. The active compounds of the present invention also may be veterinary compositions having a therapeutically effective amount of an aromatic compound of the general formula provided above in combination with a non-formula (I) active agent, optionally included in a pharmaceutically-acceptable carrier. The term "pharmaceutically-acceptable carrier" as used herein means one or more compatible solid or 30 liquid filler, dilutants or encapsulating substances which are suitable for administration to a human or other animal. The term "carrier" denotes an organic or inorganic ingredient, natural or synthetic, with which the active ingredient is combined to facilitate the application. The WO 99/26628 PCT/US98/24967 -42 components of the pharmaceutical compositions also are capable of being commingled with the compound of the present invention, and with each other, in a manner such that there is no interaction which would substantially impair the desired pharmaceutical efficacy. A common administration vehicle (e.g., pill, tablet, bolus, powder or solution for 5 dilution, pig pump, implant, injectable solution, etc.) would contain both the compounds useful in this invention and a non-formula (I) active agent. Thus, the present invention provides pharmaceutical or veterinary compositions, for medical or veterinary use, which comprise the active compounds of the invention together with one or more pharmaceutically acceptable carriers thereof and other therapeutic ingredients. 10 A variety of administration routes are available. The particular mode selected will depend of course, upon the particular drug selected, the severity of the condition being treated and the dosage required for therapeutic efficacy. The methods of this invention, generally speaking, may be practiced using any mode of administration that is medically acceptable, meaning any mode that produces effective levels of the active compounds without causing 15 clinically unacceptable adverse effects. Such modes of administration include oral, rectal, topical, nasal, transdermal or parenteral routes. The term "parenteral" includes subcutaneous, intravenous, intramuscular, or infusion. Intravenous and intramuscular routes are not particularly suited for long term therapy and prophylaxis. They could, however, be preferred in emergency situations. Oral administration will be preferred for prophylactic treatment 20 because of the convenience to the subject as well as the dosing schedule. The compositions may conveniently be presented in unit dosage form and may be prepared by any of the methods well known in the art of pharmacy. All methods include the step of bringing the active compounds into association with a carrier which constitutes one or more accessory ingredients. In general, the compositions are prepared by uniformly and 25 intimately bringing the active compounds into association with a liquid carrier, a finely divided solid carrier, or both, and then, if necessary, shaping the product. Compositions suitable for parenteral administration conveniently comprise a sterile aqueous preparation of the active compound, which is preferably isotonic with the blood of the recipient. This aqueous preparation may be formulated according to known methods 30 using those suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally-acceptable diluent or solvent, for example as a solution in 1, 3-butane diol.

WO 99/26628 PCT/US98/24967 - 43 Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil may be employed including synthetic mono or di-glycerides. In addition, fatty acids 5 such as oleic acid find use in the preparation of injectables. Carrier formulations suitable for oral, subcutaneous, intravenous, intramuscular, etc. can be found in Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, PA. Other delivery systems can include time-release, delayed release or sustained release delivery systems. Such systems can avoid repeated administrations of the active compounds 10 of the invention, increasing convenience to the subject and the physician. Many types of release delivery systems are available and known to those of ordinary skill in the art. They include polymer based systems such as polylactic and polyglycolic acid, polyanhydrides and polycaprolactone; nonpolymer systems that are lipids including sterols such as cholesterol, cholesterol esters and fatty acids or neutral fats such as mono-, di- and triglycerides; hydrogel 15 release systems; silastic systems; peptide based systems; wax coatings, compressed tablets using conventional binders and excipients, partially fused implants and the like. Specific examples include, but are not limited to: (a) erosional systems in which an agent of the invention is contained in a form within a matrix such as those described in U.S. Patent Nos. 4,452,775, 4,675,189, and 5,736,152, and (b) diffusional systems in which an active 20 component permeates at a controlled rate from a polymer such as described in U.S. Patent Nos. 3,854,480, 5,133,974 and 5,407,686. In addition, pump-based hardware delivery systems can be used, some of which are adapted for implantation. Use of a long-term sustained release implant may be particularly suitable for treatment of diarrhea in immunodeficient subjects, who need continuous administration of the 25 compositions of the invention. "Long-term" release, as used herein, means that the implant is constructed and arranged to deliver therapeutic levels of the active ingredient for at least 30 days, and preferably 60 days. Long-term sustained release implants are well known to those of ordinary skill in the art and include some of the release systems described above.

WO 99/26628 PCT/US98/24967 -44 Effective Dosages Pharmaceutical compositions suitable for use with the present invention include compositions wherein the active ingredient is contained in a therapeutically effective amount, i.e., in an amount effective to achieve its intended purpose. Of course, the actual amount 5 effective for a particular application will depend, inter alia, on the condition being treated. For example, when administered in methods to reduce sickle cell dehydration and/or delay the occurrence of erythrocyte sickling or distortion in situ, such compositions will contain an amount of active ingredient effective to achieve this result. When administered in methods to inhibit cell proliferation, such compositions will contain an amount of active ingredient 10 effective to achieve this result. When administered to subjects suffering from disorders characterized by abnormal cell proliferation, such compositions will contain an amount of active ingredient effective to, inter alia, prevent the development of or alleviate the existing symptoms of, or prolong the survival of, the subject being treated. For use in the treatment of cancer, a therapeutically effective amount further includes that amount of compound which 15 arrests or regresses the growth of a tumor. Determination of an effective amount is well within the capabilities of those skilled in the art, especially in light of the detailed disclosure herein. For any compound described herein the therapeutically effective amount can be initially determined from cell culture assays. Target plasma concentrations will be those 20 concentrations of active compound(s) that are capable of inducing at least about 25% inhibition of the Gardos channel, at least about 25% inhibition of Cl- secretion in intestinal cells and/or at least about 25% inhibition of cell proliferation in cell culture assays, depending, of course, on the particular desired application. Target plasma concentrations of active compound(s) that are capable of inducing at least about 50%, 75%, or even 90% or 25 higher inhibition of the Gardos channel, of Cl- secretion in intestinal cells, and/or of cell proliferation in cell culture assays are preferred. The percentage inhibition of the Gardos channel, of Cl- secretion in intestinal cells, and/or cell proliferation in the subject can be monitored to assess the appropriateness of the plasma drug concentration achieved, and the dosage can be adjusted upwards or downwards to achieve the desired percentage of inhibition. 30 Therapeutically effective amounts for use in humans can also be determined from animal models. For example, a dose for humans can be formulated to achieve a circulating concentration that has been found to be effective in animals. Useful animal models for WO 99/26628 PCT/US98/24967 -45 diseases characterized by abnormal cell proliferation are well-known in the art. A particularly useful animal model for sickle cell disease is the SAD mouse model (Trudel et al., 1991, EMBO J. 11:3157-3165). Useful animal models for diseases characterized by abnormal cell proliferation are well-known in the art. In particular, the following references provide suitable 5 animal models for cancer xenografts (Corbett et al., 1996, J. Exp. Ther. Oncol. 1:95-108; Dykes et al., 1992, Contrib. Oncol. Basel. Karger 42:1-22), restenosis (Carter et al., 1994, J. Am. Coll. Cardiol. 24(5):1398-1405), atherosclerosis (Zhu et al., 1994, Cardiology 85(6):370 377) and neovascularization (Epstein et al., 1987, Cornea 6(4):250-257). The dosage in humans can be adjusted by monitoring Gardos channel inhibition and/or inhibition of cell 10 proliferation and adjusting the dosage upwards or downwards, as described above. Additional in vivo assays are well known in the art. For instance, the following assays are useful for assessing effective amounts of compounds for treating inflammatory diseases associated with cellular proliferation: Airway inflammation and hyperresponsiveness in Ovalbumin-sensitized mice or guinea pigs; NZB/NZW crossed mice develop glomerular 15 disease and lupus-like syndrome; Renal allograft rejection in mice; Trinitrobenzene sulphonic acid induced bowel inflammation in rats; NZB/NZW crossed mice develop glomerular disease and lupus-like syndrome; Experimental allergic encephalomyelitis; Rat adjuvant arthritis assay; HLA transgenic mice immunized with thyroglobulin; and Thiouracil-fed rats. A therapeutically effective dose can also be determined from human data for 20 compounds which are known to exhibit similar pharmacological activities, such as Clotrimazole and other antimycotic agents (see, e.g., Brugnara et al., 1995, JPET 273:266 272; Benzaquen et al., 1995, Nature Medicine 1:534-540; Brugnara et al., 1996, J. Clin. Invest. 97(5):1227-1234). The applied dose can be adjusted based on the relative bioavailability and potency of the administered compound as compared with Clotrimazole. 25 Adjusting the dose to achieve maximal efficacy in humans based on the methods described above and other methods as are well-known in the art is well within the capabilities of the ordinarily skilled artisan. Of course, in the case of local administration, the systemic circulating concentration of administered compound will not be of particular importance. In such instances, the compound 30 is administered so as to achieve a concentration at the local area effective to achieve the intended result. For use in the prophylaxis and/or treatment of sickle cell disease, including both WO 99/26628 PCT/US98/24967 - 46 chronic sickle cell episodes and acute sickle cell crisis, a circulating concentration of administered compound of about 0.001 pM to 20 pM is considered to be effective, with about 0.1 pM to 5 .M being preferred. Subject doses for oral administration of the compounds described herein, which is the 5 preferred mode of administration for prophylaxis and for treatment of chronic sickle cell episodes, typically range from about 80 mg/day to 16,000 mg/day, more typically from about 800 mg/day to 8000 mg/day, and most typically from about 800 mg/day to 4000 mg/day. Stated in terms of subject body weight, typical dosages range from about 1 to 200 mg/kg/day, more typically from about 10 to 100 mg/kg/day, and most typically from about 10 to 50 10 mg/kg/day. Stated in terms of subject body surface areas, typical dosages range from about 40 to 8000 mg/m 2 /day, more typically from about 400 to 4000 mg/m 2 /day, and most typically from about 400 to 2000 mg/m 2 /day. For use in the treatment of disorders characterized by abnormal cell proliferation, including cancer, arteriosclerosis and angiogenic conditions such as restenosis, a circulating 15 concentration of administered compound of about 0.001 pM to 20 pM is considered to be effective, with about 0.1 pM to 5 pM being preferred. Subject doses for oral administration of the compounds described herein for the treatment or prevention of cell proliferative disorders typically range from about 80 mg/day to 16,000 mg/day, more typically from about 800 mg/day to 8000 mg/day, and most typically 20 from about 800 mg/day to 4000 mg/day. Stated in terms of subject body weight, typical dosages range from about 1 to 200 mg/kg/day, more typically from about 10 to 100 mg/kg/day, and most typically from about 10 to 50 mg/kg/day. Stated in terms of subject body surface areas, typical dosages range from about 40 to 8000 mg/m 2 /day, more typically from about 400 to 4000 mg/m 2 /day, and most typically from about 400 to 2000 mg/m 2 /day. 25 For other modes of administration, dosage amount and interval can be adjusted individually to provide plasma levels of the administered compound effective for the particular clinical indication being treated. For example, if acute sickle crises are the most dominant clinical manifestation, a compound according to the invention can be administered in relatively high concentrations multiple times per day. Alternatively, if the subject exhibits 30 only periodic sickle cell crises on an infrequent or periodic or irregular basis, it may be more desirable to administer a compound of the invention at minimal effective concentrations and to use a less frequent regimen of administration. This will provide a therapeutic regimen that WO 99/26628 PCT/US98/24967 - 47 is commensurate with the severity of the sickle cell disease state. For use in the treatment of tumorigenic cancers, the compounds can be administered before, during or after surgical removal of the tumor. For example, the compounds can be administered to the tumor via injection into the tumor mass prior to surgery in a single or 5 several doses. The tumor, or as much as possible of the tumor, may then be removed surgically. Further dosages of the drug at the tumor site can be applied post removal. Alternatively, surgical removal of as much as possible of the tumor can precede administration of the compounds at the tumor site. Combined with the teachings provided herein, by choosing among the various active 10 compounds and weighing factors such as potency, relative bioavailability, subject body weight, severity of adverse side-effects and preferred mode of administration, an effective prophylactic or therapeutic treatment regimen can be planned which does not cause substantial toxicity and yet is entirely effective to treat the clinical symptoms demonstrated by the particular subject. Of course, many factors are important in determining a therapeutic 15 regimen suitable for a particular indication or subject. Severe indications such as cancer may warrant administration of higher dosages as compared with less severe indications such as sickle cell disease. The formulations of the invention are also administered in effective amounts when treating diarrhea or scours. An effective amount is one sufficient to inhibit or prevent diarrhea 20 or scours and is thus sufficient to inhibit the Cl- secretion of intestinal epithelial cells. An amount which is sufficient to inhibit the Cl- secretion of intestinal epithelial cells thereby effectively decreases the secretory response, thereby resulting in a decrease in diarrhea or scours and/or the symptoms thereof. Effective amounts will depend, of course, on the particular condition being treated; the severity of the condition; individual subject parameters 25 including age, physical condition, size and weight; concurrent treatment; frequency of treatment; and the mode of administration. These factors are well known to those of ordinary skill in the art and can be addressed with no more than routine experimentation. An effective amount for an individual compound may be assesed using any method known in the art which reliably determines the amount of C- secretion from intestinal cells. 30 A compound may be subject to a series of standard assays or screens to determine its pharmacological activity and effective amounts. In general, the active compounds of the invention are those which induce at least about WO 99/26628 PCT/US98/24967 -48 25% inhibition of the Cl- secretion, as measured using in vitro assays that are commonly known in the art (see, e.g., Example 4). Alternatively, or in addition, the active compounds of the invention generally will have an IC 5 0 (concentration of compound that yields 50% inhibition) for inhibition of the Cl- secretion of less than about 10 pM as measured using in 5 vitro assays. It is preferred generally that a maximum dose be used, that is, the highest safe dose according to sound medical judgment, particularly if acute diarrhea or scours are the dominant clinical manifestation. Dosage may be adjusted appropriately to achieve desired drug plasma levels. Generally, daily oral doses of active compounds will be from about 0.01 10 milligrams/kg per day to 1000 milligrams/kg per day. It is expected that oral doses in the range of 50 to 500 milligrams/kg, in one or several administrations per day, will yield the desired results. In the event that the response in a subject is insufficient at such doses, even higher doses (or effective higher doses by a different, more localized delivery route) may be employed to the extent that subject tolerance permits. Multiple doses per day are 15 contemplated to achieve appropriate systemic levels of compounds. Toxicity The ratio between toxicity and therapeutic effect for a particular compound is its therapeutic index and can be expressed as the ratio between LD 5 0 (the amount of compound lethal in 50% of the population) and ED 5 0 (the amount of compound effective in 50% of the 20 population). Compounds which exhibit high therapeutic indices are preferred. Therapeutic index data obtained from cell culture assays and/or animal studies can be used in formulating a range of dosages for use in humans. The dosage of such compounds preferably lies within a range of plasma concentrations that include the ED 50 with little or no toxicity. The dosage may vary within this range depending upon the dosage form employed and the route of 25 administration utilized. The exact formulation, route of administration and dosage can be chosen by the individual physician in view of the subject's condition. (See e.g. Fingl et al., 1975, In: The Pharmacological Basis of Therapeutics, Ch. 1 p1). The invention having been described, the following examples are intended to illustrate, not limit, the invention. Some of the following examples depict tests that are employed to 30 determine the effects on Cl- secretion. Clotrimazole, which is outside of the scope of the present claims is used to exemplify how the compounds of the present invention are tested for certain conditions. The compounds of the present invention are structurally distinct from the WO 99/26628 PCT/US98/24967 - 49 structure of clotrimazole. Nevertheless, the compounds of the present invention act on chloride secretion in the same manner as clotrimazole and, therefore, are useful in the methods and products of the present invention. 5 Examples Example 1: Compound Syntheses This Example demonstrates general methods for synthesizing the compounds of the invention, as well as preferred methods for synthesizing certain exemplary compounds of the invention. In all the reaction schemes described herein, suitable starting materials are either 10 commercially available or readily obtainable using standard techniques of organic synthesis. Where necessary, suitable groups and schemes for protecting the various functionalities are well-known in the art, and can be found, for example, in Kocienski, Protecting Groups, Georg Thieme Verlag, New York, 1994 and Greene & Wuts, Protective Groups in Organic Chemistry, John Wiley & Sons, New York, 1991. 15 In FIGS. 1 and 2, the various substituents are as defined for structure (I). 1.1. Synthesis of 11 -Aryl-5,6-dihydro- 11H-dibenz[b.e] azepines This example provides a general method for synthesizing substituted 11-aryl-5,6 dihydro-11H-dibenz[b,e]azepine compounds according to the invention. A general reaction scheme is provided in FIG. 1. In FIG. 1, R 2

-R

,5 are as previously defined for structural 20 formula (I). Referring to FIG. 1, a mixture of an appropriately substituted 2-aminobenzophenone 100 (1 equivalent), an appropriately substituted benzyl chloride 102 (1 equivalent), potassium carbonate (2 equivalents) and sodium iodide (1 equivalent) in acetonitrile is refluxed for 12 hours. The reaction mixture is cooled to room temperature and water added. The mixture is 25 extracted with ethyl acetate. The combined ethyl acetate extracts are washed with water then dried over sodium sulfate. Evaporation of the solvent followed by column chromatography gives the substituted N-alkyl-2-aminobenzophenone derivative 104 in about 55-80% yield. The substituted N-alkyl-2-aminobenzophenone derivative 104 (1 equivalent) is dissolved in a 3:1 mixture of tetrahydrofuran:methanol. Sodium borohydride (10 equivalents) 30 is slowly added and the reaction mixture is stirred at room temperature for 12 hours. The reaction is quenched by adding 2 N aqueous hydrochloric acid solution. The reaction mixture is neutralized by adding 4 N aqueous sodium hydroxide solution and extracted with ethyl WO 99/26628 PCT/US98/24967 - 50 acetate. The combined ethyl acetate extracts are dried over sodium sulfate. Evaporation of the solvent followed by column chromatography gives the substituted N-alkyl-2-amino benzyl alcohol derivative 106 in about 40-60% yield. A mixture of the substituted N-alkyl-2-amino-benzylalcohol derivative 106 (1 5 equivalent), phosphorous pentoxide (5 equivalents) and methanesulfonic acid (5 equivalents) in dichloromethane is stirred at room temperature for 12 hours. The mixture is neutralized by adding aqueous sodium carbonate and then extracted with dichloromethane. The organic solution is dried over sodium sulfate. Evaporation of the solvent followed by column chromatography gives the substituted 11 -aryl-5,6-dihydro-11H-dibenz[b,e]azepine derivative 10 108 in about 45-70% yield. The substituted 11-aryl-5,6-dihydro- 11H-dibenz[b,e]azepine derivative 108 (1 equivalent) is combined with potassium carbonate (3.5 equivalents) and alkyl or acyl halide (3 equivalents) in acetonitrile and stirred at room temperature for two days. Water is added and the mixture is stirred for 15 min. at room temperature and extracted with ethyl acetate. 15 Evaporation of the solvent gives the crude product as an oil. Tituration of the product from ethanol followed by washing with hexane gives the pure N-substituted 11-aryl-5,6-dihydro 11H-dibenz[b,e]azepine product 110 as a white solid in 30-80% yield. Alternatively, the substituted 11-aryl-5,6-dihydro-11 H-dibenz[b,e]azepines can be synthesized from appropriate starting materials according to the methods described in 20 Sasakura and Sugasawa, 1981, Heterocycles 15:421-425. 1.2. Synthesis of 11 -Arvl- 11-substituted-5,6-dihydro-dibenz[b.e]azepines This example provides a general method for synthesizing 11-aryl- 11-substituted-5,6 dihydro-dibenz[b,e]azepine compounds according to the invention. A general reaction 25 scheme is provided in FIG. 2. In FIG. 2, R 1

-R

1 5 are as previously defined for structural formula (I). Referring to FIG. 2, the substituted N-alkyl-2-aminobenzophenone derivative 104 is prepared as described in Section 1.1, supra. To a solution (0.25 M) of an approriate grignard reagent in diethyl ether at -40oC is added a solution (0.1 M) of the substituted N-alkyl-2 30 aminobenzophenone derivative 104 in diethyl ether. The mixture is stirred at -40 0 C for 30 min., warmed to room temperature and quenched with water. Extraction with ethyl acetate and evaporation of the solvent gives the crude alcohol product 112 as an oil. The alcohol WO 99/26628 PCT/US98/24967 -51 product 112 is purified by column chromatography to give the pure alcohol product 112 as a white solid in 85-90% yield. Compound 112 (1 equivalent) is dissolved in dichloromethane to 0.5-1.0 M. Phosphorous pentoxide (4 equivalents) and methane sulfonic acid (4 equivalents) are added 5 and the mixture is stirred at room temperature for 2 hours. The reaction is quenched with saturated aqueous sodium bicarbonate and extracted with ethyl acetate. Evaporation of the solvent followed by column chromatography gives the 11 -aryl-11-substituted-5,6-dihydro dibenz[b,e]azepine 114 in about 90% yield. The 11-aryl- 11-substituted-5,6-dihydro-dibenz[b,e]azepine 114 can be converted to 10 the corresponding N-substituted-11-aryl-11-substituted-5,6-dihydro-dibenz[b,e]azepine 116 as described in Section 1.1, supra. 1.3. Synthesis of N-Methoxycarbonvl -11-(2'-chlorophenvl)-5,6-dihydro- 11H dibenz[be]azepine (Compound 9) 15 A preferred method of synthesis of N-methoxycarbonyl -11-(2'-chlorophenyl)-5,6 dihydro-11H-dibenz[b,e]azepine (Compound 9) is as follows: A mixture of 0.3 g (0.00098 mole) of 11-(2'-chlorophenyl)-5,6-dihydro-11H-dibenz[b,e]azepine, 1.08 g (0.0078 mole) of potassium carbonate and 1.54 g (0.016 mole) of methyl chloroformate in 10 mL of acetonitrile, was refluxed for 12 hours. The mixture was then allowed to cool to room 20 temperature and stirred with 15 mL of water for 10 minutes. The reaction mixture was extracted with ethyl acetate (2 x 15 mL). The organic layer was dried over magnesium sulfate. Evaporation gave the crude product as a brown solid. Trituration of the crude product with ethanol and washing the obtained solid with hexane gave 0.172 g (48% yield) of a white solid having a melting point of 159-161 'C. 25 The product gave the following analytical data: NMR (CDC1 3 ): 6 3.10 ppm (3H, s,

OCH

3 ); 6 4.45 ppm (1H, d, J=10 Hz, CH 2 N); 6 5.48 ppm (1H, s, CH); 6 5.82 ppm (1H, d, J=10 Hz, CH 2 N); 6 6.94 ppm (1H, m, aryl); 6 7.10 ppm (4H, m, aryl); 6 7.28 ppm (6H, m, aryl); 6 7.69 ppm (1H, m, aryl). 30 1.4. Synthesis ofN-Phenoxycarbonvl-11-phenvl-5,6-dihydro-11H-dibenz[b.e]azepine (Compound 14) A preferred method of synthesis of N-phenoxycarbonyl- 11-phenyl-5,6-dihydro-11H- WO 99/26628 PCT/US98/24967 - 52 dibenz[b,e]azepine (Compound 14) is as follows: A mixture of 0.25 g (0.00092 mole) of 11 phenyl-5,6-dihydro- 11H-dibenz[b,e]azepine, 0.318 g (0.0023 mole) of potassium carbonate and 0.318 g (0.002 mole) of phenyl chloroformate in 10 mL of acetonitrile, was stirred at room temperature for 2 days. The mixture was stirred with 15 mL of water for 15 minutes 5 and then extracted with ethyl acetate (2 x 35 mL). The organic layer was dried over magnesium sulfate. Evaporation gave the crude product as an oily material. Trituration of the obtained oil with ethanol then washing it with hexane gave 0.285 g (80% yield) of a white solid having a melting point of 155-165oC. The product gave the following analytical data: NMR (CDC1 3 ): 8 4.46 ppm (1H, d, 10 J=7 Hz, CH 2 N); 6 5.28 ppm (1H, s, CH); 6 5.69 ppm (1H, d, J =7 Hz, CH 2 N); 8 6.52 ppm (2H, m, aryl); 6 6.98 ppm (2H, m, aryl); 6 7.14 - 7.42 ppm (13H, m, aryl); 6 7.58 (1H, m, aryl). 1.5. Synthesis of N-Phenoxycarbonvl- 11 -(2'-chlorophenvyl)-5.6-dihydro- 11 H 15 dibenz[b.e]azepine (Compound 26) A preferred method of synthesis of N-phenoxycarbonyl- 11-(2'-chlorophenyl)-5,6 dihydro-11H-dibenz[b,e]azepine (Compound 26) is as follows: A mixture of 0.2 g (0.00065 mole) of 11-(2'-chlorophenyl)-5,6-dihydro- 11H-dibenz[b,e]azepine, 0.18 g (0.0013 mole) of potassium carbonate and 0.204 g (0.0013 mole) of phenyl chloroformate in 10 mL of 20 acetonitrile, was stirred at room temperature for 2 days. The mixture was stirred with 15 mL of water for 10 minutes and then extracted with ethyl acetate (2 x 35 mL). The organic layer was dried over magnesium sulfate, filtered and the solvent was evaporated. Trituration of the obtained residue with ethanol then washing it with hexane gave 0.082 g (30 % yield) of a white solid having a melting point of 95-99 0 C. 25 The product gave the following analytical data: NMR (CDC13): 6 4.50 ppm (1H, d, J=7 Hz, CH 2 N); 6 5.58 ppm (1H, s, CH); 6 5.80 ppm (1H, d, J=7 Hz, CH 2 N); 6 6.52 ppm (2H, m, aryl); 6 7.06 - 7.38 ppm (14H, m, aryl); 6 7.79 ppm (1H, m, aryl). 30 1.6. Synthesis of N-(4'-Nitrobenzoyl)-11-(2'-chlorophenvl)-5,6-dihydro-11H dibenz[b.e]azepine (Compound 28) A preferred method of synthesis of N-(4'-nitrobenzoyl)-I 1-(2'-chlorophenyl)-5,6- WO 99/26628 PCT/US98/24967 - 53 dihydro-11H-dibenz[b,e]azepine (Compound 28) is as follows: A mixture of 0.2 g (0.00065 mole) of 11-(2'-chlorophenyl)-5,6-dihydro-11H-dibenz[b,e]azepine, 0.179 g (0.0013 mole) of potassium carbonate and 0.133 g (0.00072 mole) of 4-nitrobenzoyl chloride in 10 mL of acetonitrile, was stirred at room temperature for 12 hours. The mixture was stirred with 15 5 mL of water for 10 minutes. The reaction mixture was extracted with ethyl acetate (2 x 15 mL). The organic layer was dried over magnesium sulfate. Evaporation gave the crude product as a sticky solid. Trituration of the crude product with ethanol and washing the obtained solid with hexane gave 0.148 g (50% yield) of a white solid having a melting point of 178-181oC. 10 The product gave the following analytical data: NMR (CDC13): 6 4.42 ppm (1H, d, J=7 Hz, CH 2 N); 6 5.68 ppm (1H, s, CH); 6 6.36 ppm (1H, d, J=7 Hz, CH 2 N); 8 6.52 ppm (3H, m, aryl); 6 7.06 ppm (3H, m, aryl); 6 7.12 ppm (1H, m, aryl); 5 7.26 ppm (6H, m, aryl); 6 7.79 ppm (3H, m, aryl). 15 1.7. Other Compounds Other compounds of the invention can be synthesized by routine modification of the above-described syntheses, or by other methods that are well known in the art. Appropriate starting materials are commercially available or can be synthesized using routine methods. 20 Example 2: In Vitro Activity This Example demonstrates the ability of several exemplary compounds of formula (I) to inhibit the Gardos channel of erythrocytes (Gardos channel assay) and/or mitogen-induced cell proliferation (mitogenic assay) in vitro. The assays are generally applicable for demonstrating the in vitro activity of other compounds of formula (I). 25 Methods. The percent inhibition of the Gardos channel (10 pM compound) and the ICs 0 were determined as described in Brugnara et al., 1993, J. Biol. Chem. 268(12):8760 8768. The percent inhibition of mitogen-induced cell proliferation (10 tM compound) and the IC 5 0 were determined as described in Benzaquen et al. (1995, Nature Medicine 1:534 540) with NIH 3T3 mouse fibroblast cells (ATCC No. CRL 1658). Other cell lines, e.g., 30 cancer cells, endothelial cells and fibroblasts, as well as many others, may be used in the cell proliferation assay. Selection of a particular cell line will depend in part on the desired application, and is well within the capabilities of an ordinarily skilled artisan.

WO 99/26628 PCT/US98/24967 - 54 Results. The results of the experiment are provided in Table 2, below. Clotrimazole is reported for purposes of comparison. Most of the compounds tested exhibited significant activity in both assays. All of the compounds tested exhibited significant activity in at least one of the assays. 5 Table 2 IN VITRO DATA FOR EXEMPLARY COMPOUNDS Mitogenic Assay Gardos Channel Assay 10 Compound IC 50 so Inhibition (%) IC 5 0 Inhibition (%) (RM) (IM) Clotrimazole 0.626 93.0 0.046 99.3 (1) 56.0 0.775 75.2 (2) 5.20 99.0 1.30 99.0 (3) 2.40 99.0 0.886 97.4 15 (4) 1.5 89.0 0.384 98.1 (5) 91.0 >10.0 14.4 (6) 87.0 0.236 97.5 (7) 1.60 99.0 >10.0 35.8 (8) 2.20 84.0 0 20 (9) 2.10 99.0 0.0850-0.093 97.3 (10) 53.0 1.533-1.940 63.0 (11) 32.0 >10.0 9.5 (12) 13.0 >10.0 54.8 (13) 1.7 97.0 0 25 (14) 0.04 98.0 >10.0 14.8 (15) 40.0 >10.0 9.50 (16) 1.7 99.0 >10.0 0.45 (17) 1.6 99.0 >10.0 20.6 (18) 2.6 99.0 0.502-0.692 81.5 30 (19) 1.6 99.0 >10.0 52.0 (20) 1.7 95.0 >10.0 13.6 (21) 2.7 93.0 >10.0 2.1 (22) 3.6 99.0 >10.0 14.9 (23) 55.0 >10.0 18.2 WO 99/26628 PCT/US98/24967 - 55 Mitogenic Assay Gardos Channel Assay O10 Compound IC 50 so Inhibition (%) IC 50 so Inhibition (%) (AM) (AM) (24) 89.0 >10.0 32-55 (25) 75.0 >10.0 8.5 (26) 0.04-0.90 99.0 >10.0 0.8 (27) 2.20 99.0 >10.0 3.0 5 (28) 0.04-0.50 99.0 0 (29) 0.800 99.0 0.414-0.433 95.1 (30) 0.600 99.0 >10 14.6 (31) 0.400 99.0 >10 12.3 (32) 1.100 99.0 0 10 (33) 2.400 99.0 >10 67.5 (34) 4.00 99.0 >10 12.0 (35) 0 0.071-0.099 98.3 Example 3: Activity In Cancer Cell Lines 15 This Example demonstrates the antiproliferative effect of several exemplary compounds of formula (I) against a variety of cancer cell lines. The assays are generally applicable for demonstrating the antiproliferative activity of other compounds of formula (I). Methods. Growth of Cells. The antiproliferative assays described herein were performed using standard aseptic procedures and universal precautions for the use of tissues. 20 Cells were propagated using RPMI 1640 media (Gibco) containing 2% or 5% fetal calf serum (FCS) (Biowhittaker) at 37 0 C, 5% CO 2 and 95% humidity. The cells were passaged using Trypsin (Gibco). Prior to addition of test compound, the cells were harvested, the cell number counted and seeded at 10,000 cells/well in 100 pl 5% fetal calf serum (FCS) containing RPMI medium in 96-well plates and incubated overnight at 37oC, 5% CO 2 and 95% humidity. 25 On the day of the treatment, stock solutions of the test compounds (10 mM compound/DMSO) were added in 100 pil FCS containing medium to a final concentration of 10-0.125 pM and the cells were incubated for 2, 3 or 5 days at 37oC, 5% CO 2 and 95% humidity. Following incubation, the cellular protein was determined with the Sulforhodamine B 30 (SRB) assay (Skehan et al., 1990, J. Natl. Cancer Inst. 82:1107-1112). Growth inhibition, WO 99/26628 PCT/US98/24967 - 56 reported as the concentration of test compound which inhibited 50% of cell proliferation

(IC

5 0 ) was determined by curve fitting. Values for VP-16, a standard anti-cancer agent, are provided for comparison. Except for MMRU cells, all cancer cell lines tested were obtained from the American 5 Type Culture Collection (ATCC, Rockville, MD). The ATCC assession numbers were as follows: HeLa (CCL-2); CaSki (CRL-1550); MDA-MB-231 (HTB-26); MCF-7 (HTB-22); A549 (CCL-185); HTB-174 (HTB-174); HEPG2 (HB-8065); DU-145 (HTB-81); SK-MEL 28 (HTB-72); HT-29 (HTB-38); HCT-15 (CCL-225); ACHN (CRL-1611); U-118MG (HTB 15); SK-OV-3 (HTB-77). 10 MMRU cells (Stender et al., 1993, J. Dermatology 20:611-617) were a gift of one of the authors. Results. The results of the cell culture assays are presented in Tables 3 and 4, below.

WO 99/26628 57- PCTIUS98/24967 m 00'C' 0 r C l A r 0 r - l 1 - Cl ( N ' C> C) AN C Cl C:> (= 00 C) 0 0 - 00 en 0In 00 6 6 6 6 6 6 An A 0 CD 6 6 6 C0 AC> 6 A A A A e~ 0 0 A 00 o ,- 0 -I- 0 0 A 0 A con 71 00 CN cnC A~ A -c C> C) I C :)C17 It A( C 11 ON ) ON ' 0 040 0q C! .N .0 . \ N - ~ 00 'N 0( - C r- N -I ON '(N 0 >( 0 WO 99/26628 -58 -PCT/US98/24967 - V V Qall -v A A A - A A fU 0D 0) Cl Cl Cl 0 Cl C AI tN 10 10 ~- 0 0Cl 00 0 C CDON O kn- A A 0 A A ~*) ~-. ~ ~ 0~~~~ ~ ~ L C6 -V) ~ - - ~ - ~ ~ Qz ml Cl 'Nt 1/N m/ 1/N 1/N alN MI 1/ / /N 'N U on -n Ln Nn 00 'n Ln -n 'n 'n Lnn r m hI O N-z ~ f 0 If N 0 11N0 WO 99/26628 PCT/US98/24967 -59 As can be seen in Tables 3 and 4, compounds which exhibited significant activity in the mitogenic assay described in Section 7, supra, exhibit significant antiproliferative activity against a variety of cancer cell lines and cancer types (IC 5 0 of less than about 10 tM). Many of the compounds exhibit comparable or even greater antiproliferative activity 5 against a variety of cancer cell types than VP- 16, a known anti-cancer agent. Example 4: Formulations The following examples provide exemplary, not limiting, formulations for administering the compounds of the invention to mammalian, especially human, patients. 10 Any of the compounds described herein, or pharmaceutical salts or hydrates thereof, may be formulated as provided in the following examples. 4.1. Tablet Formulation Tablets each containing 60 mg of active ingredient are made up as follows: 15 Active Compound 150 mg Starch 150 mg Microcrystalline Cellulose 150 mg Sodium carboxymethyl starch 4.5 mg Talc 1 mg 20 Polyvinylpyrrolidone (10% in 4 mg water) Magnesium Stearate 0.5 mg 160 mg 25 The active ingredient, starch and cellulose are passed through a No. 45 mesh U.S. sieve and mixed thoroughly. The solution of polyvinylpyrrolidone is mixed with the resultant powders which are then passed through a No. 14 mesh U.S. sieve. The granules are dried at 50o-60'C and passed through a No. 18 mesh U.S. sieve. The sodium carboxymethyl starch, magnesium stearate and talc, previously passed through a No. 60 mesh U.S. sieve, are then 30 added to the granules, which, after mixing are compressed by a tablet machine to yield tablets each weighing 150 mg. Tablets can be prepared from the ingredients listed by wet granulation followed by compression.

WO 99/26628 PCT/US98/24967 - 60 4.2. Gelatin Capsules Hard gelatin capsules are prepared using the following ingredients: Active Compound 250 mg/capsule 5 Starch dried 200 mg/capsule Magnesium Stearate 10 mg/capsule The above ingredients are mixed and filled into hard gelatin capsules in 460 mg quantities. 10 4.3. Aerosol Solution An aerosol solution is prepared containing the following components: Active Compound 0.25% (w/w) Ethanol 29.75% (w/w) 15 Propellant 22 77.00% (w/w) (Chlorodifluoromethane) The active compound is mixed with ethanol and the mixture added to a portion of the propellant 22, cooled to -30 0 C and transferred to a filling device. The required amount is then 20 fed to a stainless steel container and diluted with the remainder of the propellant. The valve units are then fitted to the container. 4.4. Suppositories Suppositories each containing 225 mg of active ingredient are made as follows: 25 Active Compound 225 mg Saturated fatty acid glycerides 2,000 mg The active ingredient is passed through a No. 60 mesh U.S. sieve and suspended in the 30 saturated fatty acid glycerides previously melted using the minimum heat necessary. The mixture is then poured into a suppository mold of nominal 2 g capacity and allowed to cool.

WO 99/26628 PCT/US98/24967 -61 4.5. Suspensions Suspensions each containing 50 mg of medicament per 5 mL dose are made as follows: 5 Active Compound 50 mg Sodium carboxymethylcellulose 50 mg Syrup 1.25 mL Benzoic acid solution 0.10 mL Flavor q.v. 10 Color q.v. Purified water to 5 mL The active ingredient is passed through a No. 45 mesh U.S. sieve and mixed with the sodium carboxymethyl cellulose and syrup to form a smooth paste. The benzoic acid 15 solution, flavor and some color are diluted with some of the water and added, with stirring. Sufficient water is then added to produce the required volume. Example 5: Clotrimazole inhibits water and electrolyte secretion in intestinal epithelial cells. 20 The biochemical basis of secretory diarrhea involves intestinal Cl- secretion in intestinal crypt cells. Under normal conditions, Cl- ions are maintained within intestinal crypt cells at levels above their electrochemical potential by primarily and secondarily active transport mechanisms such as the Na/K ATPase pumps and Na/K/2Cl cotransporters. Cl- is transported into the lumen from the intestinal crypt cells through apical Cl- channels. 25 Intracellular levels of K*, cAMP, cGMP, and Ca ++ are all involved in regulating the secretory response. T84 cells were used to determine whether clotrimazole regulates Cl- secretion in intestinal crypt cells. T84 cells form confluent monolayers of columnar epithelia that exhibit high transepithelial resistances, polarized apical and basilateral membranes, and cAMP and 30 Ca ++ regulated Cl- secretory pathways analogous to those found in native intestine. Methods. Growth of T84 cells. T84 cells obtained from ATCC were cultured and passaged in equal parts of dulbecco's modified eagle's medium (DMEM), 1 g/1 D-glucose) and Hams F-12 nutrient mixture, supplemented with 5% newborn calf serum, 15 mM HEPES, 14 mM Na HCO 3 , 40mg/1 penicillin, 8mg/1 ampicillin, 0.90 mg/1 streptomycin. Cells were 35 seeded at confluent density onto 0.33 cm 2 or 5cm 2 Transwell inserts (Costar, Cambridge, MA) WO 99/26628 PCT/US98/24967 - 62 coated with dilute rat collagen solution as previously described (Lencer et al., J. Clin. Invest., 92: 2941-2951 (1993); Lencer et al., J. Cell Biol. 117: 1197-1209 (1992). Transepithelial resistances attain stable levels (>1000 Ohms.cm 2 ) after 7 days. The development of high transepithelial resistances correlated with the formation of confluent monolayers with well 5 developed tight junctions as assessed by morphological analysis, and with the ability of monolayers to secrete Cl- (Madara et al., Gastro. 92: 1133-1145 (1987). Electrophysiology (mesurement of electrogenic Cl- secretion). Confluent monolayers were transferred to Hanks Buffered Salt Solution (HBSS) containing 0.185 g/1 CaCl 2 , 0.098 g/1 MgSO 4 , 0.4 g/l KC1, 0.06 g/l KH 2

PO

4 , 8 NaC1, 0.048 g/l Na 2

HPO

4 , 1 g/l glucose, and 10 10OnM HEPES, pH 7.4. Serosal and mucosal reservoirs were interfaced with Calomel and Ag Ag Cl electrodes via 5% agar bridges made with Ringer's buffer. Transepithelial resistance was measured using a dual voltage clamp device to apply 25 or 50pA current pulses. Short circuit current (ISC) was calculated using Ohms law as previously described (Lencer et al., J. Clin. Invest. 92: 2941-2951 (1993); Lencer et al. J. Cell Biol. 117: 1197-1209 (1992). 15 Results: Clotrimazole reversibly inhibits Cl- secretion elicited by Ca"- or cAMP dependant agonists in T84 cells. Previous studies have shown that Cl- secretion in T84 cells is controlled by K' efflux pathways which are biophysically and pharmacologically distinct from one another. One pathway participates in the secretory response to cAMP-dependent agonists and displays sensitivity to Ba" salts (McRoberts, et al., J. Biol. Chem. 260: 14163 20 14172 (1985); Reenstra, Am J Physiol. 264: C161-168 (1993)). The other mediates the response to Ca+-dependent agonists, and is Ba"-insensitive. Several pathway specific agonists of K+ channels are useful for determining whether a particular compound is functioning through a cAMP or Ca specific pathway. For instance, vasoactive intestinal peptide (VIP) and cholera toxin are cAMP mediated agonists of the K' channel, whereas, 25 carbachol is a Ca+-dependent agonist of the Ca" regulated K' channels. The pathway by which a particular inhibitor of Cl- secretion in T84 cells is functioning may be identified by measuring the ability of the inhibitor to modify transepithelial resistances in T84 cells which have been treated with VIP or carbachol to stimulate Cl- secretion. T84 cells were grown as described above and C1- secretion was stimulated by the 30 addition to the media of either carbachol (100mM) or VIP (5nM). The cells were then treated with BaCl (3mM), charybdotoxin (100nM), or clotrimazole (33mM). The short circuit current (ISC) was determined for the various inhibitor treatments as a percentage of the control in the WO 99/26628 PCT/US98/24967 - 63 absence of inhibitor (FIG. 3). BaCl strongly inhibited the secretory response to the cAMP mediated agonist VIP, but had no apparent affect on the secretory response elicited by the Ca"-dependent agonist carbachol. In contrast, the scorpion venom Charybdotoxin strongly inhibited the secretory response elicited by carbachol, but had minimal affects on Cl- secretion 5 elicited by VIP. However, clotrimazole inhibited the Cl secretory responses to both agonists. Inhibition of Cl- secretion by clotrimazole was fully reversible (96+2%, n = 4) after 60 min recovery in the presence of 0.01 mg/ml bovine serum albumin. To examine possible effects of clotrimazole on the synergy between cAMP and Ca

+

"

mediated agonists, monolayers, initially stimulated with VIP were allowed to reach steady 10 state levels of secretion and then additionally exposed to carbachol (100 pM). Clotrimazole was slightly more effective in inhibiting the secretory response to carbachol than to cAMP with IC50 values of 3 and 8 pM, respectively. When the effects of clotrimazole on cAMP and Ca+-dependent secretory pathways were examined on the same monolayers., inhibition of the synergistic response to VIP plus carbachol was found to parallel the inhibition of secretion 15 promoted by Ca" agonists alone. In low doses (=10 7 or less), clotrimazole potentiated slightly (by 5-10%) the CI-secretory responses to either agonist. clotrimazole inhibited effectively the secretory response to cholera toxin (20 nM, a cAMP-dependent agonist) and E Coli heat-stabile toxin (100 nm, a cGMP-agonist) (IC50 values of 10 pM and 15 pM, respectively). 20 The effect of clotrimazole on K conductances was also examined by isotopic flux studies using 86RB. T84 cells were grown in the presence of a cAMP agonist, VIP, or a Ca" mediated agonist (Thapsigargin). Clotrimazole was added and 8 6 RB efflux was measured. Clotrimazole significantly inhibited baseline and Ca" stimulated 86RB efflux in the presence of both cAMP and Ca" mediated agonists compared to those cells which were not treated 25 with clotrimazole. Other aromatic compunds of the invention were found to inhibit chloride secretion. Although clotrimazole was the most potent inhibitor tested of cAMP and Ca" elicited Cl secretion, ketoconazole, econazole, miconazole, and 2-chlorophenyl-bis-phenyl methanol also were effective at inhibiting chloride secretion. 30 Taken together, these studies indicate that clotrimazole inhibits Cl- secretion elicited by cAMP or Ca

+

mediated K' channels in T84 cells.

WO 99/26628 PCT/US98/24967 - 64 Example 6: Clotrimazole acts at distal steps in the cAMP and Ca++-dependent signal transduction pathways. To determine the site of clotrimazole action, the effects of clotrimazole pretreatment were examined on monolayers stimulated with agonists that initiate Cl- secretion at sequential 5 steps in the cAMP signalling cascade. T84 monolayers were preincubated in HBSS in the presence or absence of clotrimazole (33 pM) and then stimulated with either 5 pM VIP (which activates adenylate cyclase through heterotrimeric GTPase-linked cell surface receptors), 10 p-M forskolin (which activates adenylate cyclase directly), or 3 mM 8Br-cAMP (a direct stimulator of protein kinase A). Clotrimazole inhibited the secretory response to 10 each of these agonists. These data provide evidence that clotrimazole acts at a step distal to the activation of Protein Kinase A. Ca+-dependent intracellular signaling in T84 and other non-exciteable cells involves recruitment of inositol trisphosphate (IP3)-dependent intracellular Ca++ stores (Halm and Frizzell, Textbook of Secretory Diarrhea, Raven Press, 47-58 (1990); Mandel et al., J Biol. 15 Chem. 267: 704-712 (1986); Halm et al., Am. J. Physiol. (Cell Physiol. 23) 254:C505-C511 (1988)), and subsequent activation of plasma membrane Ca++ influx pathways (Barrett, Am. J. Physiol. (Cell Physiol. 34): C859-C868 (1993)). Downstream events may be mediated by [Ca++]i, IP3, diacylglycerol, or as yet unidentified diffusable factors (Putney and Bird, Cell 75:199-201 (1993)). To examine the site of clotrimazole action alone, this signalling, 20 cascade, T84 monolayers pretreated in the presence or absence of clotrimazole (33 ptM) were stimulated with the Ca++-dependent agonists carbachol (100 p-M which elicits both Ca++ and

IP

3 signals), thapsigargin (5 p-M, which elevates cytoplasmic Ca++ via inhibition of ER Ca++ ATPase) (Vandorpe et al., Biophys. J 66:46-58 (1994)), or the Ca++ ionophore ionomycin (10 p-M). Clotrimazole inhibited strongly the Cl-secretory response to each to these reagents. 25 These data suggest that clotrimazole acts at steps in the secretory response distal to the release of intracellular Ca++ stores. Example 7: Clotrimazole does not affect apical membrane anion conductance or basolateral NaK2Cl cotransporters. 30 Methods: 25I Efflux Studies. Confluent monolayers on 5 cm 2 Transwell inserts were used 10-14 days after plating. 1251 was measured as an indicator of apical Cl-, channel and basolateral K+ channel activity as previously described (Venglarik, et al, Am. J Physiol. (Cell WO 99/26628 PCT/US98/24967 - 65 Physiol. 28):C358-C364 (1990). Monolayers were preincubated at 37' C with 4 pCi/ml 1251 in HBSS for 90 minutes, with 33 pM clotrimazole absent or present during the final 30 minutes of this 90 min preincubation period. Clotrimazole pretreatment did not alter 1251 loading of the cells. After washing twice in fresh HBSS, 0.5 ml samples were obtained every 5 two min from the apical reservoir and replaced with fresh HBSS. After four baseline samples were obtained, the cells were treated (at t = 8 minutes) with vasoactive intestinal peptide (VIP, 5 pnM) or thapsigargin (5 tM) to stimulate Cl- secretion, and an additional 15 timed samples were obtained. Finally, the cell monolayer was rinsed, cut with its support from the polystyrene ring, and the residual cell-associated radioactivity was determined. Monolayers 10 were maintained at 370 C in room air throughout the study. 1251I was counted by gamma counting and normalized to percent total uptake as previously described (Venglarik, et al, Am. J Physiol. (Cell Physiol. 28):C358-C364 (1990). MRb Uptake Studies. Confluent monolayers on 5 cm 2 Transwell inserts were incubated for 30 minutes in HBSS at 37' C. A group of control and CLT treated (33 tM, for 15 30 min) monolayers were treated with bumetanide (10 pM for 12 min). All monolayers were then treated with VIP (5nM and shifted to HBSS containing 1 ICi/ml 86 Rb for 3 minutes at 37 C. 8 6 Rb uptake was terminated by washing the inserts in an ice-cold solution containing 100mM MgCI 2 , and 10mM TRIS-CL, pH 7.4. Monolayers were cut from their inserts, placed into scintillation vials, and counted using standard methods. 20 Results: Studies were conducted to determine whether the inhibition of electrogenic CI- secretion might occur by blockade of apical membrane Cl-channels, or blockade of basolaterally situated NaK2Cl cotransporters. To determine if clotrimazole affected ion conductance through apical membrane Cl-channels, we examined the time course of 1251 efflux from T84 monolayers pretreated in the presence or absence of clotrimazole (Venglarik, et al, 25 Am. J. Physiol. (Cell Physiol. 28):C358-C364 (1990). Clotrimazole had little or no effect on the time course of 125I efflux from monolayers treated with VIP. Rate constants for 125I efflux from monolayers treated or not treated with clotrimazole were indistinguishable (0.0637 vs.0.0645 % uptake/minute, n=2 in duplicate). Clotrimazole had similar lack of effect on 1251 efflux stimulated by thapsigargin. 30 We next tested the effect of clotrimazole on basolateral NaK2CI cotransporters, as assessed by bumetanide-sensitive 86 Rb uptake (Matthews et al., J Biol. Chem. 269:15703 15709 (1994)). Clotrimazole treatment reduced the total amount of 8 6 Rb uptake by 53.6+5.8% WO 99/26628 PCT/US98/24967 - 66 (mean±SEM. n=6), but had no effect on the fractional component that was bumetanide sensitive (88±3.2 vs 75.2 ±12.7% total uptake, meaniSEM). Taken together, these data strongly suggest that clotrimazole does not affect CI- secretion in T84 cells via inhibition of either apical membrane Cl- channels or basolateral membrane NaK2Cl cotransporters. 5 Example 8: Clotrimazole inhibits Chloride secretion by inhibiting K+ efflux through basolateral K+ channels in T84 cells. 8.1. Clotrimazole inhibits chloride secretion by blockade of K+ transport through both Ba++ 10 sensitive and charybdotoxin-sensitive channels. Methods: 86 Rb Efflux Studies Confluent monolayers on 5 cm 2 Transwell inserts were used 10-14 days after plating. 86 Rb flux was measured as an indicator of apical Cl-, channel and basolateral K+ channel activity as previously described (Venglarik, et al, Am. J. Physiol. (Cell Physiol. 28):C358-C364 (1990). Monolayers were preincubated at 370 C with 4 ptCi/ml 15 8 6 Rb in HBSS for 90 minutes, with 33 pM clotrimazole absent or present during the final 30 minutes of this 90 min preincubation period. clotrimazole pretreatment did not alter 8 6 Rb loading of the cells. One ml samples were obtained and replaced from the basolateral reservoir. After four baseline samples were obtained, the cells were treated (at t = 8 minutes) with vasoactive intestinal peptide (VIP, 5 pnM) or thapsigargin (5 tM) to stimulate Cl 20 secretion, and an additional 15 timed samples were obtained. Finally, the cell monolayer was rinsed, cut with its support from the polystyrene ring, and the residual cell-associated radioactivity was determined. Monolayers were maintained at 37 oC in room air throughout the study. 8 6 Rb was counted by scintillation counting and normalized to percent total uptake as previously described (Venglarik, et al, Am. J. Physiol. (Cell Physiol. 28):C358-C364 25 (1990). Results: K+ channel activity was estimated by measurement of 6 Rb efflux. Clotrimazole was found to significantly inhibit the rate of " 6 Rb efflux after treatment with the cAMP agonist VIP (5 tM). The rate constant for VIP-stimulated 86 Rb efflux was reduced by 87% in monolayers treated with clotrimazole (0.0062 vs. 0.0465 % uptake/minute, n=2 in 30 triplicate). clotrimazole inhibited to a similar degree 86 Rb efflux from monolayers stimulated with thapsigargin (panel B, rate constants 0.011 vs. 0.048% uptake/minute, n=2), suggesting that clotrimazole can inhibit Cl- secretion by blockade of K+ transport through both Ba++- WO 99/26628 PCT/US98/24967 - 67 sensitive and charybdotoxin-sensitive channels. 8.2. Clotrimazole inhibits chloride secretion through distinct cAMP and Ca- sensitive basolateral K' channels. 5 Methods: Selective membrane Permeabilization and Measurement of Potassium Conductance of the Basolateral Membrane. The basolateral potassium conductance was measured using the technique developed by Dawson and co-workers. A potassium gradient (mucosal to serosal) was first established across the monolayer using asymmetric mucosal and serosal buffers containing K' as the sole permeant ion. The addition of amphotericin B (20 10 pM) to the mucosal reservoir forms conductive pores in the apical membrane, and thus removes all resistance to transepithelial potassium movement across this membrane. Thus, under the conditions of the experiment, in which the monolayer is short circuited (i.e., voltage-clamped at zero potential) and the transepithehal potassium gradient is constant, the amphotericin-dependent Isc becomes a measure of the rate of the transepithelial potassium 15 flux across basolateral membranes. Changes in short circuit current (Isc), then represent changes in basolateral K' conductances (gK). Isc and K' conductances were measured using calomel electrodes, 3M KCI-agar bridges, and a voltage clamp (University of Iowa, Iowa City). To generate a voltage-current channel relationships, currents were elicited by 1 sec test potentials from -80 to +80 in 10 mV increments in the asymmetrical high K* gluconate 20 solution. Calculation of basolateral membrane K permeability. Membrane permeabilities were calculated according to the formula: PK= (cm/s)=JK (mM/cm2*s)/A[ K

+

] (mM/cm 3 ) where A [K ] is equal to the difference in K' concentration (135 mM) between the asymmetric 25 apical and basolateral bathing solutions. Maximal Isc values were converted into K+ fluxes by dividing by the Faraday constant F (96,500 coulombs/mol) as previously described (Huflejt et al., J. Clin. Invest. 93: 1900-1910 (1994)). Results: Basolateral K+ transport was examined in T84 monolayers permeabilized apically by pretreatment with amphotericin B. Apical and basolateral buffers contained K+ as 30 the sole permeant ion. All studies were performed with a 135 mM basolaterally directed K+ gradient. This method has been utilized previously to examine both Cl- and K+ transport in T84 cells and HT29-C1.16E cells. Briefly, ion conductances in the luminal or basolateral WO 99/26628 PCT/US98/24967 - 68 membranes of confluent T84 cell monolayers can be assessed separately by selectively permeabilizing the apical or basolateral membrane using the ionophore amphotericin B. This artificially removes all electrical resistance to ion transport across the plasma membrane containing pores formed by amphotericin B. As a result, the intact contralateral plasma 5 membrane becomes rate limiting for transepithelial ion transport. Agonist-dependent changes in ion conductances can be assessed directly either as transepithelial short circuit current (Isc) in the presence of established ion gradients, or as transepithelial conductance (G) in the presence of established transepithelial potentials. K+ transport was measured at baseline and after the ordered additions of cAMP- and 10 Ca' -agonists. The initial permeabilization with amphotericin B was associated with 49 ± 19% increase in conductance. Pores formed by amphotericin B display selectivity for monovalent cations. Ca" remained relatively impermeant as evidenced by the small steady state increase in Isc and GK caused by apical permeabilization with amphotericin B. Given this low baseline Isc and GK, both cAMP- and Ca+-sensitive K+ permeabilities (PK) were 15 readily apparent after agonist stimulation. Treatment with the cAMP-agonist forskolin (10 iM) caused a brisk increase in K+ transport through apparently low-conductance pathway(s), as evidenced by symmetrical increases in Isc and G. Carbachol also increased K+ currents. The magnitude of the carbachol-induced IscK, however, was similar whether carbachol was added alone or after forskolin (111.7 ± 7.4 vs. 180.7 ± 15.7tA/cm 2 respectively. Thus, there 20 was no clear evidence of synergy between cAMP and Ca + mediated K+ pathways, as would be expected in an apically permeabilized cell system. Analagous to our previous findings in intact T84 monolayers, the forskolin-induced changes in Isc were sustained while the effect of carbachol was short-lived. Both IscK and GK returned to baseline values within 5 min after addition of carbachol. 25 Formal current/voltage (I/V) relations were defined before and after agonist stimulation to confirm that both cAMP- and Ca -dependent currents were elicited at physiologic membrane potentials. Thapsigargin was used in place of carbacol as a Ca

+

agonist in these studies because the K+ transients elicited by thapsigargin achieve steady state conductances of much longer duration, as in intact monolayers. It was found that under 30 conditions of basolaterally directed K+ gradients, both forskolin and thapsigargin activate macroscopic outwardly rectified (mucosal to serosal) currents at positive transepithelial voltages. Experimental I/V relations obtained after forskolin and thapsigargin stimulation WO 99/26628 PCT/US98/24967 - 69 displayed reversal potentials (- 40 mV) that approximated the calculated Nernst-potential (-85 mV calculated as RT/zQo log [Na]out/[Na]in). These results are consistent with the activation of distinct cAMP- and Ca+-sensitive basolateral membrane K+ conductances in conjunction with one or more nonspecific transepithelial ion shunts, possibly occurring through 5 intercellular tight junctions or basolateral membrane "leaks." To confirm that the observed changes in Isc and G represented K+ transport through K+ selective pathways, the effect of forskolin and carbachol on T84 monolayer conductances were examined using buffers containing Na as the sole permeant cation. These studies were performed using an analogous 135 mM basolaterally directed cation (Na

+

) gradient. Increases 10 in Isc and G were not detectable in the absence of K+. Thus, the increases in cation conductances induced by agonist stimulation are specific to K+ transport. Two pharmacologically distinct K+ efflux pathways have been previously identified in intact T84 cells. One pathway participates in the secretary response to cAMP-dependent agonists and displays sensitivity to Ba + salts. The other K+ efflux pathway mediates the 15 response to Ca+-dependent agonists, and is Ba+-insensitive. These findings were confirmed in the permeabolized cell model. The cAMP-sensitive IK (elicited by treatment with forskolin, 10 [M) was inhibited by greater than 70% by the addition of BaC12 (3 mM) to basolateral reservoirs. Ba + , however, had no detectable effect on K+ transport induced by the subsequent addition of carbachol (100 pM) to the same monolayers. In contrast, when 20 permeabilized monolayers were treated first with carbachol, the induced Ca* IK was inhibited by 50% by pretreatment with the scorpion venom charybdotoxin (100 nM). Charybdotoxin, however, had no detectable effect on K+ transport induced by the subsequent addition of forskolin. Thus in permeabilized cells, the differential sensitivity of K+ transport to inhibition by the K+ channel blockers BaCl 2 and charybdotoxin paralleled exactly the effect of these 25 channel selective inhibitors on K+ transport in intact cells (measured indirectly as a Cl current). Taken together, these studies define the permeabilized T84 cell model, and provide strong evidence that under the defined conditions both Isc and G represent K+ transport through distinct cAMP- and Ca+-sensitive basolateral K+ channels. 30 8.3. Clotrimazole and 2-chlorophenvl-bs-phenvl methanol, a structurally related stable metabolite. inhibit K+ transport through both cAMP- and Ca+-dependent K+ channels.

WO 99/26628 PCT/US98/24967 - 70 We next tested the hypothesis that clotrimazole may inhibit directly basolateral membrane K+ channels in human intestinal T84 cells, as it does in the red cell. clotrimazole significantly inhibited the time course of K+ transport after treatment with the cAMP agonist forskolin (10 pM) and the Ca + agonist carbachol (100 pM). Formal IV relations taken at 5 steady state after cAMP or Ca' stimulation confirm that clotrimazole affected both cAMP and Ca"- sensitive channels. Nearly identical results were obtained with 2-chlorophenyl-bis phenyl methanol. clotrimazole and its metabolite 2-chlorophenyl-bis-phenyl methanol inhibit directly both cAMP- and Ca+-sensitive intestinal K+ channels indicating that the ring structure in the absence of the imidazole ring sufficient (and perhaps necessary) for this 10 bioactivity. 8.4. Clotrimazole targets the basolateral rather than the apical surface of T84 cells Methods: Measurement of Cl Conductance of the Apical Plasma Membrane. To examine apical Cl conductances, Cl- was used as the sole permeant ion using identical apical 15 and basolateral buffer solutions. Monolayers were pemeabilized basolaterally by the addition of 100 pM Amphotericin B to the serosal reservoir. Generation of voltage-current curves of channel currents were elicited by 1 sec test potentials from -80 to + 80 mV in 10 mV increments in symmetrical high Choline Cl- buffers. Results: Studies were performed to determine whether the primary target of 20 clotrimazole was located on the basoolateral or apical cell surfaces. Most rapid inhibition was achieved by incubation with clotrimazole on both sides of the monolayer. However, basolateral application alone was almost as effective as incubation on both sides. Additionally, the apparent potency of inhibition of clotrimazole at a fixed time point was found to be greater when applied basolaterally than apically. This preferential action of 25 clotrimazole at the basolateral surface of the cell is consistent with the hypothesis that its principal targets are basolateral K+ channels. To confirm these findings, we examined Cl- transport in T84 cell monolayers permeabilized basolaterally with pores formed by amphotericin B. These studies were performed with Cl- as the only permeant anion, and with symmetrical apical and basolateral 30 Cl- concentrations (142 mM). In monolayers not treated with clotrimazole, the addition of forskolin (10 pM) to basolateral reservoirs increased Cl- conductances significantly over baseline, presumably via activation of the cystic fibrosis transmembrane regulator (CFTR) Cl- WO 99/26628 PCT/US98/24967 -71 channel. In contrast to the clear inhibitory effects of clotrimazole on basolateral K+ conductances, however, clotrimazole had no detectable effect on either forskolin- or thapsigargin-stimulated Cl-conductances. I/V relations for Cl- transport were nearly identical in monolayers treated or not treated with clotrimazole. These data provide further evidence 5 that clotrimazole inhibits Cl- secretion in intact T84 cell monolayers by affecting specifically basolateral K+ channels. Apical membrane Cl-channels are not inhibited. Example 9: Clotrimazole inhibits C1 secretion in vivo. 9.1. Chamber studies using rabbit colonic mucosa. 10 Methods: 4 male, New Zealand rabbits (2.5 kg) were anesthetized by an intravenous injection of pentobarbital (0.5 ml/kg). A 15 cm length of distal colon was removed and opened longitudinally. External muscle layers were removed by blunt dissection and colonic mucosal preparations were mounted in an Ussing chamber (DCTSYS; Precision Instrument Design, CA; 10.3 cm 2 surface area) and incubated with buffer solution containing (in mM): 15 NaCl 122.0, CaCl 2 , 2.0; MgSO 4 ,1.3; KC1, 5.0, glucose, 20; NaHCO 3 , 25.0 (pH when gassed with 95% 02/5 C,0 2 ; temperature was maintained at 37°C) with and without clotrimazole (30M). The volume of fluid on each side of the mucosa was 7 ml. Potential difference and Isc were monitored continuously and registered every 10 minutes. Luminal and serosal buffer solutions were interfaced via Ag-AgCI electrodes 20 (Voltage/Current Clamp, Model VCC600, Physiologic Instruments, Inc., San Diego, CA, USA) and Ringer/agar bridge to voltage clamp device (model DVC-1000; Voltage/Current Clamp, World Precision Instruments, Inc.). Resistance (R) was calculated using Ohm's law and the Isc and is given in O x cm2. After stable baseline resistance and Isc values had been obtained, mucosal preparations were incubated in the presence or absence of serosal 25 clotrimazole (30 rtM) for 30 min, and then stimulated by the addition of forskolin (10 tM) or carbachol (10 tM) to the serosal reservoir. Results: To test the ability of clotrimazole to block K+ channels and thus Cl- secretion in native intestinal tissue, we mounted isolated preparations of rabbit colonic mucosa in Ussing chambers containing modified Ringer's solution with or without clotrimazole (30 pM). After 30 Isc had stabilized, successive additions of forskolin (10 tM) and then cubachol (100 tM) were applied to serosal reservoirs, and Isc and G were monitored continuously. clotrimazole inhibited strongly the time course of forskolin induced Isc. Carbachol had no further effect on WO 99/26628 PCT/US98/24967 - 72 Isc in this system. 9.2. Murine model of secretotory diarrhea. Methods: Treated and control, untreated, mice were gavage fed either clotrimazole 5 (150 mg/kg/day divided in two equal doses, dissolved in peanut oil at a concentration of 20 mg/ml) or vehicle control over a 7 day loading period. Mice were then challenged by gavage with either 25 pg purified cholera toxin (Calbiochem, San Diego, CA) in PBS, vehicle control alone (PBS without cholera toxin), or cholera toxin in PBS containing 30 pM clotrimazole. Animals were sacrificed after 5 hours in an uncrowded CO 2 hood. The carcass was weighed, 10 the abdomen was opened, and ligatures were tied at the proximal duodenum and distal rectum. The intestinal block was dissected free of supporting structures and removed as a single unit and weighed. Small and large intestinal segments were normalized to body weight (intestinal weight/carcass weight) for each animal. Results: To examine whether clotrimazole may inhibit intestinal secretion in vivo, we 15 utilized a murine model of secretary diarrhea. Balb/C mice were gavage fed 150 mg/kg/day clotrimazole, divided into two equal doses, or vehicle control every 12 h for 7 days and subsequently challenged orally with purified cholera toxin (25 jg). Five hours after treatment with cholera toxin, the mice were sacrificed and intestinal fluid secretion assessed gravimetrically. Pretreatment with clotrimazole reduced by 86% intestinal fluid secretion 20 induced by cholera toxin. Clotrimazole had no effect on intestinal secretion in the absence of cholera toxin. Thus, clotrimazole effectively treated secretory diarrhea in vivo, presumably by inhibiting basolateral K+ channels of crypt epithelial cells. The foregoing written specification is considered to be sufficient to enable one skilled in the art to practice the invention. The present invention is not to be limited in scope by 25 examples provided, since the examples are intended as a single illustration of one aspect of the invention and other functionally equivalent embodiments are within the scope of the invention. The advantages and objects of the invention are not necessarily encompassed by each embodiment of the invention. Each of the foregoing patents, patent applications and references is herein incorporated 30 by reference in its entirety. What Is Claimed Is:

Claims (51)

1. A method for inhibiting unwanted cellular proliferation associated with an inflammatory disease, said method comprising the step of contacting a cell the proliferation of 5 which contributes to inflammation in situ with an effective amount of a compound having the formula: Rs R6 \15 R5 N R 7 R4 10 R8 R RR, / R3 (I)a0"a R11 "R13 R12 15 or a pharmaceutically acceptable salt or hydrate thereof, wherein: R 1 is -R', (C 6 -C 20 ) aryl or substituted (C 6 -C 20 ) aryl; 20 R, is -R', -OR', -SR', halogen or trihalomethyl; R 3 is -R', -OR', -SR', halogen or trihalomethyl or, when taken together with R 4 , is (C 6 C 20 ) aryleno; R 4 is -R', -OR', -SR', halogen or trihalomethyl or, when taken together with R 3 , is (C 6 C 2 0 ) aryleno; 25 each of Rs, R 6 , R 7 , Rs, R 9 , Rio, R 11 , R 12 , RI 3 and RI 4 is independently selected from the group consisting of -R', halogen and trihalomethyl; R, 5 is -R", -C(O)R", -C(S)R", -C(O)OR", -C(S)OR", -C(O)SR", -C(S)SR", -C(O)N(R") 2 , -C(S)N(R") 2 , -C(O)C(O)R", -C(S)C(O)R", -C(O)C(S)R", -C(S)C(S)R", -C(O)C(O)OR", -C(S)C(O)OR", -C(O)C(S)OR", -C(O)C(O)SR", -C(S)C(S)OR", 30 -C(S)C(O)SR", -C(O)C(S)SR", -C(S)C(S)SR", -C(O)C(O)N(R") 2 , -C(S)C(O)N(R") 2 , -C(O)C(S)N(R") 2 or -C(S)C(S)N(R")2 each R' is independently selected from the group consisting of -H, (C -C 6 ) alkyl, WO 99/26628 PCT/US98/24967 - 74 (C 1 -C 6 ) alkenyl and (C 1 -C 6 ) alkynyl; each R" is independently selected from the group consisting of -H, (C I-C 6 ) alkyl, (C 1 -C 6 ) alkenyl, (C 1 -C 6 ) alkynyl, (C 6 -C 20 ) aryl, (C 6 -C 20 ) substituted aryl, (C 6 -C 26 ) alkaryl and substituted (C 6 -C 26 ) alkaryl; and 5 the aryl and alkaryl substituents are each independently selected from the group consisting of-CN, -OR', -SR', -NO 2 , -NR'R', halogen, (C 1 -C 6 ) alkyl, (C 1 -C 6 ) alkenyl, (C 1 -C 6 ) alkynyl and trihalomethyl.

2. The method of Claim 1, wherein the compound is selected from the group 10 consisting of Compounds 1, 2, 3, 4, 6, 9, 18, 29, 35 and combinations thereof.

3. The method of Claim 2, wherein the compound is selected from the group consisting of Compounds 2, 3, 4, 6, 9, 29, 35 and combinations thereof. 15

4. The method of Claim 1, wherein said administration is selected from the group consisting of oral, parenteral, intravenous, subcutaneous, transdermal and transmucosal for a living human.

5. The method of Claim 1, wherein said mammalian cell is a fibrotic cell. 20

6. The method of Claim 1, wherein said mammalian cell is a lymphocyte.

7. A method of treating or preventing an inflammatory disease, said method comprising the step of administering to a subject suffering from an inflammatory disease a 25 therapeutically effective amount of a compound Ris R6 \1 R5 having the formula: R N R30R4 R, R9 R, R2 R3 30 R1o R14 R11 R13 R12 WO 99/26628 PCT/US98/24967 - 75 (I) 5 or a pharmaceutically acceptable salt or hydrate thereof, wherein: R 1 is -R', (C 6 -C 20 ) aryl or substituted (C 6 -C 20 ) aryl; R 2 is -R', -OR', -SR', halogen or trihalomethyl; 10 R 3 is -R', -OR', -SR', halogen or trihalomethyl or, when taken together with R 4 , is (C 6 C 20 ) aryleno; R 4 is -R', -OR', -SR', halogen or trihalomethyl or, when taken together with R 3 , is (C 6 C 20 ) aryleno; each of Rs, R 6 , R 7 , R

8 , R

9 , RIo, R, 1 , R 1 2 , R 13 and R 4 is independently selected from the 15 group consisting of -R', halogen and trihalomethyl; R ,5 is -R", -C(O)R", -C(S)R", -C(O)OR", -C(S)OR", -C(O)SR", -C(S)SR", -C(O)N(R") 2 , -C(S)N(R") 2 , -C(O)C(O)R", -C(S)C(O)R", -C(O)C(S)R", -C(S)C(S)R", -C(O)C(O)OR", -C(S)C(O)OR", -C(O)C(S)OR", -C(O)C(O)SR", -C(S)C(S)OR", -C(S)C(O)SR", -C(O)C(S)SR", -C(S)C(S)SR", -C(O)C(O)N(R") 2 , -C(S)C(O)N(R") 2 , 20 -C(O)C(S)N(R") 2 or-C(S)C(S)N(R")2 each R' is independently selected from the group consisting of-H, (C 1 -C 6 ) alkyl, (C 1 -C 6 ) alkenyl and (C 1 -C 6 ) alkynyl; each R" is independently selected from the group consisting of -H, (C 1 ,-C 6 ) alkyl, (C 1 -C 6 ) alkenyl, (C 1 -C 6 ) alkynyl, (C 6 -C 2 0 ) aryl, (C 6 -C 20 ) substituted aryl, (C 6 -C 26 ) alkaryl and 25 substituted (C 6 -C 26 ) alkaryl; and the aryl and alkaryl substituents are each independently selected from the group consisting of-CN, -OR', -SR', -NO 2 , -NR'R', halogen, (C 1 -C 6 ) alkyl, (Cl-C 6 ) alkenyl, (C 1 -C 6 ) alkynyl and trihalomethyl. 30 8. The method of Claim 7, wherein the compound is selected from the group consisting of Compounds 1, 2, 3, 4, 6, 9, 18, 29, 35 and combinations thereof WO 99/26628 PCT/US98/24967 - 76 9. The method of Claim 8, wherein the compound is selected from the group consisting of Compounds 2, 3, 4, 6, 9, 29, 35 and combinations thereof.

10. The method of Claim 7, wherein said inflammatory disease is diarrhea. 5

11. The method of Claim 10, wherein said diarhrea is caused by inflammatory bowel disease.

12. The method of Claim 7, wherein said inflammatory disease is an autoimmune 10 disease.

13. The method of Claim 12, wherein said autoimmune disease is lupus.

14. The method of Claim 7, wherein said inflammatory disease is 15 glomerulonephritis.

15. The method of Claim 7, wherein said administration is parenteral.

16. The method of Claim 7, wherein said administration is per oral. 20

17. The method of claim 7, wherein the inflammatory disease is selected from the group consisting of proliferative glomerulonephritis; lupus erythematosus; scleroderma; temporal arteritis; thromboangiitis obliterans; mucocutaneous lymph node syndrome; asthma; host versus graft; inflammatory bowel disease; multiple sclerosis; rheumatoid arthritis; 25 thyroiditis; Grave's disease; antigen-induced airway hyperactivity; pulmonary eosinophilia; Guillain-Barre syndrome; allergic rhinitis; myasthenia gravis; human T-lymphotrophic virus type 1-associated myelopathy; herpes simplex encephalitis; inflammatory myopathies; atherosclerosis; and Goodpasture's syndrome. 30

18. A method for treating diarrhea, comprising, administering an effective amount for inhibiting Cl- secretion of an aromatic compound to a subject, wherein the aromatic compound is selected from the group consisting of a substituted 11-phenyl-dibenzazepine, and WO 99/26628 PCT/US98/24967 - 77 analogues thereof.

19. The method of claim 18, wherein the aromatic compound is a compound having the structural formula: 5 R R6 \15 R5 N R / \R4 (I) R8 R R R2 R3 Ro R14 10 , R1 R13 R12 wherein: R, is -R', C 6 -C 2 0 ) aryl or substituted (C 6 -C 2 0 ) aryl; R 2 is -R', -OR', -SR', halogen or trihalomethyl; R 3 is -R', -OR', -SR', halogen or trihalomethyl or, when taken together with R 4 , 15 is (C 6 -C 20 ) aryleno; R 4 is -R', -OR', -SR', halogen or trihalomethyl or, when taken together with R 3 , is (C 6 -C 20 ) aryleno; each of R 5 , R 6 , R 7 , R 8 , R 9 , Rio, R 1 , R 1 2 , R 13 and RI 4 is independently selected from the group consisting of-R', halogen and trihalomethyl; R, 15 is -R", -C(O)R", -C(S)R", -C(O)OR", -C(S)OR", -C(O)SR", -C(S)SR", -C(O)N(R") 2 , -C(S)N(R") 2 , -C(O)C(O)R", -C(S)C(O)R", -C(O)C(S)R", -C(S)C(S)R", -C(O)C(O)OR", -C(S)C(O)OR", 20 -C(O)C(S)OR", -C(O)C(O)SR", -C(S)C(S)OR", -C(S)C(O)SR", -C(O)C(S)SR", -C(S)C(S)SR", -C(O)C(O)N(R") 2 , -C(S)C(O)N(R") 2 , -C(O)C(S)N(R") 2 or -C(S)C(S)N(R")2 each R' is independently selected from the group consisting of -H, (C 1 -C 6 ) alkyl, (C 1 -C 6 ) alkenyl and (C 1 -C 6 ) alkynyl; each R" is independently selected from the group consisting of -H, (C 1 -C 6 ) alkyl, (C 1 -C 6 ) alkenyl, (C 1 -C 6 ) alkynyl, (C 6 -C 20 ) aryl, (C 6 -C 20 ) substituted aryl, (C 6 25 C 26 ) alkaryl and substituted (C 6 -C 2 6 ) alkaryl; and the aryl and alkaryl substituents are each independently selected from the group consisting of -CN, -OR', -SR', -NO 2 , -NR'R', halogen, (C 1 -C 6 ) alkyl, (CI-C 6 ) alkenyl, (CI-C 6 ) alkynyl and trihalomethyl.

20. The method of claim 18, wherein the aromatic compound is selected from the 30 group consisting of aromatic compounds wherein the halogens are each independently -F, -Cl, -Br or -I. WO 99/26628 PCT/US98/24967 - 78

21. The method of claim 19, wherein the aromatic compound is administered orally.

22. The method of claim 19, wherein the subject is a human. 5

23. The method of claim 22, further comprising administering an anti-diarrheal agent to the subject.

24. The method of claim 23, wherein the anti-diarrheal agent is an oral rehydration 10 fluid.

25. The method of claim 19, wherein the aromatic compound is selected from the group consisting of compounds wherein R 1 5 is -R", -C(O)R", -C(O)OR", -C(O)N(R") 2 , -C(O)C(O)R", -C(O)C(O)OR" or -C(O)C(O)N(R") 2 . 15

26. A veterinary preparation comprising: an aromatic compound in an amount effective to inhibit scours in a subject, wherein the aromatic compound is selected from the group consisting of a substituted 11-phenyl dibenzazepine, and analogues thereof; and, 20 an anti-scours agent.

27. A veterinary preparation as in claim 26, wherein the aromatic compound is a compound having the structural formula: Rs R6 R\5 Rs N 25 R 7 R4 (I) R R R, R2 R3 Rio R 14 R11 R13 30 30 R12 wherein: R, is -R', C6-C20) aryl or substituted (C6-C20) aryl; R, is -R', -OR', -SR', halogen or WO 99/26628 PCT/US98/24967 - 79 trihalomethyl; R 3 is -R', -OR', -SR', halogen or trihalomethyl or, when taken together with R 4 , is (C 6 -C 20 ) aryleno; R 4 is -R', -OR', -SR', halogen or trihalomethyl or, when taken together with R 3 , is (C 6 -C 2 0 ) aryleno; each of Rs, R 6 , R 7 , Rs, R 9 , Rio, Ri , R]2, R 13 and R 14 is independently selected from the group consisting of-R', halogen and trihalomethyl; R 1 5 is -R", 5 -C(O)R", -C(S)R", -C(O)OR", -C(S)OR", -C(O)SR", -C(S)SR", -C(O)N(R") 2 , -C(S)N(R") 2 , -C(O)C(O)R", -C(S)C(O)R", -C(O)C(S)R", -C(S)C(S)R", -C(O)C(O)OR", -C(S)C(O)OR", -C(O)C(S)OR", -C(O)C(O)SR", -C(S)C(S)OR", -C(S)C(O)SR", -C(O)C(S)SR", -C(S)C(S)SR", -C(O)C(O)N(R") 2 , -C(S)C(O)N(R") 2 , -C(O)C(S)N(R") 2 or -C(S)C(S)N(R")2 each R' is independently selected from the group consisting of -H, (C -C 6 ) alkyl, (C 1 -C 6 ) 10 alkenyl and (C 1 -C 6 ) alkynyl; each R" is independently selected from the group consisting of -H, (C 1 -C 6 ) alkyl, (C 1 -C 6 ) alkenyl, (CI-C 6 ) alkynyl, (C 6 -C 20 ) aryl, (C 6 -C 2 0 ) substituted aryl, (C 6 C 26 ) alkaryl and substituted (C 6 -C 26 ) alkaryl; and the aryl and alkaryl substituents are each independently selected from the group consisting of -CN, -OR', -SR', -NO 2 , -NR'R', halogen, (CI-C 6 ) alkyl, (CI-C 6 ) alkenyl, (C 1 -C 6 ) alkynyl and trihalomethy. 15

28. The veterinary preparation as in claim 26, wherein the anti-scours agent is a colostral extract.

29. The veterinary preparation as in claim 26, wherein the anti-scours agent is an 20 immunological preparation of colostrum.

30. The veterinary preparation as in claim 26, wherein the anti-scours agent is a microorganism specific immunological preparation. 25

31. The veterinary preparation as in claim 26, wherein the anti-scours agent is an oral rehydration fluid.

32. The veterinary preparation as in claim 26, wherein the anti-scours agent is a replacement electrolyte composition. 30

33. The veterinary preparation as in claim 26, wherein the anti-scours agent is an antibiotic composition. WO 99/26628 PCT/US98/24967 - 80

34. The veterinary preparation as in claim 26, wherein the veterinary preparation is a dry preparation.

35. The veterinary preparation as in claim 26, wherein the aromatic compound is 5 selected from the group consisting of aromatic compounds wherein the halogens are each independently -F, -Cl, -Br or -I.

36. A pharmaceutical preparation, comprising: an aromatic compound in an amount effective to inhibit diarrhea, wherein the aromatic 10 compound is selected from the group consisting of a substituted 11-phenyl-dibenzazepine, and analogues thereof; and, an anti-diarrheal agent.

37. The pharmaceutical preparation as in claim 36, wherein the aromatic 15 compound is a compound having the structural formula: Rs R6 \1 Rs N20 20 R 7 R4 (I) R R, R2 R3 Ro R14 R11 R13 25 25 R12 wherein: R, is -R', C 6 -C 20 ) aryl or substituted (C 6 -C 20 ) aryl; R 2 is -R', -OR', -SR', halogen or trihalomethyl; R 3 is -R', -OR', -SR', halogen or trihalomethyl or, when taken together with R 4 , is (C 6 -C 20 ) aryleno; R 4 is -R', -OR', -SR', halogen or trihalomethyl or, when taken together 30 with R 3 , is (C 6 -C 2 0 ) aryleno; each of Rs, R 6 , R 7 , R 8 , R 9 , Rio, R 1 , RI 2 , R 13 and RI 4 is independently selected from the group consisting of-R', halogen and trihalomethyl; R, 5 is -R", -C(O)R", -C(S)R", -C(O)OR", -C(S)OR", -C(O)SR", -C(S)SR", -C(O)N(R") 2 , -C(S)N(R")2, WO 99/26628 PCT/US98/24967 - 81 -C(O)C(O)R", -C(S)C(O)R", -C(O)(S)CR", -C(S)C(S)R", -C(O)C(O)OR", -C(S)C(O)OR", -C(O)C(S)OR", -C(O)C(O)SR", -C(S)C(S)OR", -C(S)C(O)SR", -C(O)C(S)SR", -C(S)C(S)SR", -C(O)C(O)N(R") 2 , -C(S)C(O)N(R") 2 , -C(O)C(S)N(R") 2 or -C(S)C(S)N(R")2; each R' is independently selected from the group consisting of -H, (C -C 6 ) alkyl, (C 1 -C 6 ) 5 alkenyl and (C 1 -C 6 ) alkynyl; each R" is independently selected from the group consisting of -H, (C 1 -C 6 ) alkyl, (C 1 -C 6 ) alkenyl, (C 1 -C 6 ) alkynyl, (C 6 -C 20 ) aryl, (C 6 -C 20 ) substituted aryl, (C 6 C 26 ) alkaryl and substituted (C 6 -C 26 ) alkaryl; and the aryl and alkaryl substituents are each independently selected from the group consisting of -CN, -OR', -SR', -NO 2 , -NR'R', halogen, (CI-C 6 ) alkyl, (C 1 -C 6 ) alkenyl, (CI-C 6 ) alkynyl and trihalomethy. 10

38. The pharmaceutical preparation as in claim 37, wherein the aromatic compound is selected from the group consisting of aromatic compounds wherein the halogens are each independently -F, -Cl, -Br or -I. 15

39. The pharmaceutical preparation as in claim 37, wherein the anti-diarrheal agent is an oral rehydration fluid.

40. The pharmaceutical preparation as in claim 37, wherein the anti-diarrheal agent is an antibiotic. 20

41. The pharmaceutical preparation as in claim 37, wherein the anti-diarrheal agent is an electrolyte composition.

42. The pharmaceutical preparation as in claim 37, wherein the anti-diarrheal agent 25 is an immunoglobulin preparation from bovine colostrum.

43. The pharmaceutical preparation as in claim 37, wherein the anti-diarrheal agent is an oral sugar-electrolyte solution. 30

44. The pharmaceutical preparation as in claim 37, wherein the aromatic compound is selected from the group consisting of compounds wherein R 15 is -R", -C(O)R", -C(O)OR", -C(O)N(R") 2 , -C(O)C(O)R", -C(O)C(O)OR" or -C(O)C(O)N(R") 2 . WO 99/26628 PCT/US98/24967 - 82

45. A method for treating scours, the method comprising the step of: administering to a subject in need of such treatment, an aromatic compound in an amount effective to inhibit scours, wherein the aromatic compound is selected from the group consisting of a substituted 11-phenyl-dibenzazepine, and analogues thereof. 5

46. The method for treating scours as in claim 45, wherein the aromatic compound is a compound having the structural formula: Rs R6 R\5 Rs N R7 R4 R10 RI - I R11 R13 R12 15 wherein: R 1 is -R', C 6 -C 20 ) aryl or substituted (C 6 -C 20 ) aryl; R 2 is -R', -OR', -SR', halogen or trihalomethyl; R 3 is -R', -OR', -SR', halogen or trihalomethyl or, when taken together with R 4 , is (C 6 -C 20 ) aryleno; R 4 is -R', -OR', -SR', halogen or trihalomethyl or, when taken together with R 3 , is (C 6 -C 2 0 ) aryleno; each of Rs, R 6 , R 7 , R 8 , R 9 , R 10 , Ri 1 , RI 2 , R 3 and R 14 is 20 independently selected from the group consisting of-R', halogen and trihalomethyl; R 15 is -R", -C(O)R", -C(S)R", -C(O)OR", -C(S)OR", -C(O)SR", -C(S)SR", -C(O)N(R"') 2 , -C(S)N(R") 2 , -C(O)C(O)R", -C(S)C(O)R", -C(O)C(S)R", -C(S)C(S)R", -C(O)C(O)OR", -C(S)C(O)OR", -C(O)C(S)OR", -C(O)C(O)SR", -C(S)C(S)OR", -C(S)C(O)SR", -C(O)C(S)SR", -C(S)C(S)SR", -C(O)C(O)N(R") 2 , -C(S)C(O)N(R") 2 , -C(O)C(S)N(R") 2 or -C(S)C(S)N(R")2 25 each R' is independently selected from the group consisting of-H, (C 1 -C 6 ) alkyl, (C 1 -C 6 ) alkenyl and (C 1 -C 6 ) alkynyl; each R" is independently selected from the group consisting of -H, (C 1 -C 6 ) alkyl, (C 1 -C 6 ) alkenyl, (C 1 -C 6 ) alkynyl, (C 6 -C 20 ) aryl, (C 6 -C 20 ) substituted aryl, (C 6 C 26 ) alkaryl and substituted (C 6 -C 2 6 ) alkaryl; and the aryl and alkaryl substituents are each independently selected from the group consisting of -CN, -OR', -SR', -NO 2 , -NR'R', halogen, 30 (C 1 -C 6 ) alkyl, (CI-C 6 ) alkenyl, (CI-C 6 ) alkynyl and trihalomethy.

47. The method for treating scours as in claim 46, wherein the aromatic compound WO 99/26628 PCT/US98/24967 - 83 is selected from the group consisting of aromatic compounds wherein the halogens are each independently -F, -Cl, -Br or -I.

48. The method for treating scours as in claim 46, wherein the aromatic compound 5 is administered orally.

49. The method for treating scours as in claim 46, wherein the subject is selected from the group consisting of a horse, a cow, a pig, and a goat. 10

50. The method for treating scours as in claim 46, further comprising administering an anti-scours agent to the subject.

51. The method for treating scours as in claim 46, wherein the aromatic compound is selected from the group consisting of compounds wherein R, 5 is -R", -C(O)R", -C(O)OR", 15 -C(O)N(R") 2 , -C(O)C(O)R", -C(O)C(O)OR" or -C(O)C(O)N(R") 2 .