AU2004200272C1

AU2004200272C1 - Imidazoline Receptor binding compounds

Info

Publication number: AU2004200272C1
Application number: AU2004200272A
Authority: AU
Inventors: James E. Hall; Richard R. Tidwell; Dorothy H. Wood
Original assignee: University of North Carolina at Chapel Hill
Current assignee: University of North Carolina at Chapel Hill
Priority date: 1998-07-22
Filing date: 2004-01-23
Publication date: 2004-02-19
Anticipated expiration: 2019-06-25
Also published as: AU2004200272A1; AU2004200272B2

Description

P001 Section 29 Regulation 3.2(2)

AUSTRALIA

Patents Act 1990 COMPLETE SPECIFICATION STANDARD PATENT Application Number: Lodged: Invention Title: Imidazoline Receptor binding compounds The following statement is a full description of this invention, including the best method of performing it known to us: IMIDAZOLINE RECEPTOR BINDING COMPOUNDS Richard R. Tidwell, James E. Hall, and Dorothy H. Wood FIELD OF THE INVENTION The present invention concerns compounds that have high affinity for the imidazoline receptor, particularly the imidazoline 12 receptor, methods of use thereof, and combinatorial libraries thereof.

BACKGROUND OF THE INVENTION In the early 1980's while studying the different hypotensive effects of imidazoline compounds vs. catecholamines in the nucleus reticularis lateralis, Bousquet et al. proposed the existence of a class of imidazoline binding sites distinct from adrenergic receptors (Bousquet, P. et al., The Journal of Pharmacology and Experimental Therapeutics 230:232 236 (1984)).

Many subsequent studies have supported the presence of these binding sites in a variety of tissues (Coupry, I. et al., Biochemical Biophysical Research .Communications 147:1055 1060 (1987); Ernsberger, P. R. et al, European Journal of Pharmacology 134:1 13 (1987); Meeley, M. P. et al., Life Sciences 38:1119 1126 (1986); Parini, A. et al. Journal of Biological Chemistry 264(20):11874-8 (1989)), including the existence of the putative endogenous ligand (Atlas, and Y.

Burstein, European Journal ofBiochemistry 144:287 293 (1984); Li, G. et al., Science 263:966 969 (1994)) and purification of a mitochondrial imidazoline binding site (Limon, I. et al., Journal of Biological Chemistry 267(30):21645-9 (1992)).

The binding sites are classified into two groups, 1i and 12, according to their affinity for clonidine and idazoxan, respectively, and are located on the plasma and outer mitochondrial membranes. I2 binding sites are associated with the monoamine oxidase protein in certain tissues and are further sub-classified into 1 2A and I2B depending on their sensitivity to the compound amiloride (Regunathan, and D. J.

Reis, Annual Review ofPharmacology Toxicology 36:511-44 (1996)). A third subgroup classified as IaOpical have been identified in the pancreatic P-cell (Morgan, N.

G. et al., Annals of the New YorkAcademy of Sciences 763:361-73 (1995); Brown, C.

A. et al., British Journal of Pharmacology 108:312 317 (1993); Chan, S. Clinical Science 85(6):671-7 (1993)).

The I1 binding sites are localized on the plasma membrane and have been found in the brainstem, kidneys, adrenal chromaffin cells, rat PC12 cells and platelets.

Imidazoline II receptor agonists which stimulate I receptors in the medulla offer a new therapeutic approach to the tratinent of hypertension and associated metabolic syfidrome comprising of hyperglycemia, hyperinsulinemia, hypenriglyceridemia and low concentrations of high-density lipoprotein cholesterol (Krentz, A. and A. J.

Evans, The Lancet 351:152 153 (1998)). Selectivity for this receptor reduces the adverse effects attributed to stimulation of a2-adrenoceptors such as sedation and dry mouth. Platelet II receptors are upregulated in patients suffering with depression and can be downregulated with antidepressant therapy.

12 binding sites are more widely distributed than I sites. They have been found in the nervous system (cerebral cortex, astrocytes, carotid bodies), specific organs (kidneys, liver, pancreas, colon, urethra, placenta), glands (adrenal medulla, prostate) and certain cell types (platelets, adipocytes, vascular cells). Expression of 12 binding sites is upregulated in human brains during aging, and in patients with Alzheimer's disease. Expression is modified in the brains of depressed suicide victims (Regunathan, and D. J. Reis, Annual Review of Pharmacology Toxicology 36:511-44 (1996)).

Efforts have been made to identify compounds with imidazoline receptor binding affinity: U.S. Patent No. 5,726,197 to Clark et al. describe isoindolinyl derivatives that have imidazoline receptor binding affinity.

U.S. Patent No. 5,686,477 to Jarry et al. describes various (aryloxymethyl) oxazolines having imidazoline binding activity.

U.S. Patent No. 5,354,769 to Garcia Sevilla et al. describes benzofuranyl imidazole derivatives having imidazoline receptor binding activity, and various uses thereof.

IND

O U.S. Patent No. 5,732,717 to Watanabe et al. describes the use of 4chloro-5-(imidazoline-2-ylamino)-6-methoxy-2-methylpyrimidine for treating o substance abuse withdrawal.

O

SU.S. Patent No. 5,574,059 to Regunathan et al. describes the treatment of vascular smooth muscle cell proliferation with certain imidazoline receptor agonists.

Nevertheless, there are relatively few compounds with imidazoline receptor Sbinding affinity available, and there is continued interest in identifying additional imidazoline receptor binding compounds.

SUMMARY OF THE INVENTION The present invention relates, in a first aspect, to a method for treating Alzheimer's disease in a subject in need of treatment thereof, the method including administering to the subject a therapeutic amount of a bis-benzene compound, wherein said bis-benzene compound includes at least one amidine group, or a pharmaceutically acceptable salt thereof. Such compounds may, for example, include a compound of formula A B "PX-(CH 2

(I)

R, R 3 wherein: A and B are each independently selected from the group consisting of H, loweralkyl, oxyalkyl, nitro, amino, aminoalkyl, halo, hydroxy, carboxy, and compounds of formula

RTN

(i)

RTN

R,

subject to the proviso that at least one of A and B is a compound of formula R, and R 2 are each independently selected from the group consisting of H, loweralkyl, oxyalkyl, alkoxyalkyl, cycloalkyl, aryl, hydroxyalkyl, aminoalkyl, and alkylaminoalkyl;

IND

O or two R, groups on the same compound of formula together represent

-(CH

2 wherein m is 2, 3, or 4; o R 3 is H, loweralkyl, oxyalkyl, alkoxyalkyl, hydroxyalkyl, cycloalkyl, aryl, aminoalkyl, alkylaminoalkyl, or halogen; n is an integer from 2 to 6; and X is O, NH, or S; or a pharmaceutically acceptable salt thereof.

0 In a second aspect, the present invention provides a method for treating diabetes in a subject in need of treatment thereof, the method including administering to the subject a therapeutic amount of a bis-benzene compound, wherein said bis-benzene compound includes at least one amidine group, or a pharmaceutically acceptable salt thereof. Such compounds may also, for example, include a compound of formula A further aspect of the present invention is a method of binding the imidazoline receptor, comprising contacting a bis-benzene to said imidazoline receptor in an amount effective to bind to said receptor, wherein said bis-benzene contains at least one amidine group. The amidine group is covalently bound to one of the benzene rings, or an amidine group is covalently bound to both of the benzene rings (such compounds and their pharmaceutically acceptable salts are referred to as "active agents" herein). The contacting step may be carried out in vitro with cells that express the imidazoline receptor, or with a cell-free preparation comprising the imidazoline receptor) or in vivo by administering said compound to a subject afflicted with a disease state which is alleviable by treatment with a compound having high selectivity and affinity for the imidazoline receptor site).

A still further aspect of the present invention is a method of identifying imidazoline receptor binding agents, comprising the steps of providing a library of bis-benzene compounds, said bis-benzene compound containing at least one amidine group as described above; and screening said library for compounds that bind to said imidazoline receptor. The library may be a combinational library, and the bis-benzene compounds may be immobilized on a solid support in accordance with known techniques where only one of the benzene groups

ID

o has an amidine group covalently bound thereto, the other benzene group may be affixed, by covalent bond, to the solid support).

o The present invention is explained in greater detail in the drawings and specification set forth below.

R

1 and R 2 are each independently selected from the group consisting of H, loweralkyl, oxyalkyl, alkoxyalkyl, cycloalkyl, aryl, hydroxyalkyl, aminoalkyl and alkylaminoalkyl; or two R 1 groups on the same amidine group together represent S- (CH 2 wherein m is 2, 3, or 4; (-i

R

3 is H, loweralkyl, oxyalkyl, alkoxyalkyl, hydroxyalkyl, cycloalkyl, aryl, aminoalkyl, alkylaminoalkyl or halogen; n is from 2 to 6; and X is O, NH, or S; or a pharmaceutically acceptable salt thereof. In one embodiment, R 1

R

2 and R 2 are H; X is 0; and n is A second group of bis-benzenes that may be used to carry out the present invention are those having formula II: A N N B NH HN

(II)

wherein: A and B are each independently selected from the group consisting of H, loweralkyl, oxyalkyl, nitro, amino, aminoalkyl, halo, hydroxy, carboxy, and compounds of formula

R-

1

N

R--N R1--N R2 (i) 3c

IND

0 subject to the proviso that at least one of A and B is a compound of formula

R

1 and R 2 are each independently selected from the group consisting of H, O loweralkyl, oxyalkyl, alkoxyalkyl, cycloalkyl, aryl, hydroxyalkyl, aminoalkyl and Salkylaminoalkyl; or two R 1 group on the same amidine group together represent

(CH

2 )m-wherein m is 2, 3, or 4; cN R 3 is H, loweralkyl, oxyalkyl, alkoxyalkyl, hydroxyalkyl, cycloalkyl, aryl, Saminoalkyl, alkylaminoalkyl or halogen; BRIEF DESCRIPTION OF THE DRAWINGS Fig.l. Inhibition of 5 nM 3 H]idazoxan (Kd 14 2nM) binding to rat liver membranes by a2-adrenoceptor ligands and pentamidine. Cirazoline, Ki 3.0 0.4 nM; Phentolamine, Ki 5000 nM; Epinephrine, Ki 25,000 nM; Yohimbine, Ki 25,000 nM; Pentamidine, Ki 1.4 0.22 Nm.

DETAILED DESCRIPTION OF THE INVENTION Definitions. As used herein: The term "loweralkyl," as used herein, refers to C 1

C

6 linear or branched alkyl, such as methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, iso-butyl, tert-butyl, pentyl, isopentyl, and hexyl. Isoalkyl groups, such as isopropyl, isobutyl. isopentyl, and the like are currently preferred. The term "loweralkoxy" or."oxyalkyl" as used herein, refers to C, C 6 linear or branched alkoxy, such as methoxy, ethoxy, propyloxy, butyloxy, isopropyloxy,and t-butyloxy. Methoxy is currently preferred.

"Cycloalkyl" means a saturated monovalent monocyclic hydrocarbon radical containing 3-8 carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl.

The term "halo" or "halogen" as used herein refers halogens such as to Cl, Br, I, F, preferably Cl or Br.

"Imidazoline receptor" as used herein refers to any imidazoline receptor, including Il and 12 receptors. The 12 receptors (including 1 2A and 12B receptors) are currently preferred. The imidazoline receptor may be of any species of origin, but is preferably mammalian human, mouse, rat, cat, dog, rabbit, hamster, monkey, etc.).

"Phenyl" as used herein encompasses all possible isomeric phenyl radicals optionally monosubstituted or disubstituted with a substituent selected from the group consisting of lower alkyl, lower alkoxy, and halo.

"Pharmaceutically acceptable acid addition salt" refers to those salts which retain the biological effectiveness and properties of the free bases and which are not biologically or otherwise undesirable, formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like, and organic acids such as acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, malic acid, malonic acid, succinic acid. maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid and the like.

H]-idazoxan" refers to tritiated idazoxan, named as benzodioxan-2-yl)-2-imidazoline hydrochloride, or [3 H]-RX 781094. It is available from Amersham plc.

The term "treatment" as used herein covers any treatment of a disease in a mammal, particularly a human, and includes: preventing the disease from occurring in a subject which may be predisposed to the disease but has not yet been diagnosed as having it; (ii) inhibiting the disease, arresting its development; or io (iii) relieving the disease, causing regression of the disease.

The term "disease state which is alleviable by treatment with a compound having high selectivity and/or affinity for the imidazoline receptor site" as used herein is intended to cover all disease states which are generally acknowledged in the art to be usefully treated with compounds having high selectivity and affinity for the imidazoline site in general, as described in U.S. Patent No. 5,726,197 to Clark (see column 4, lines 1- 19 therein). Such disease states include, but are not limited to, cerebral ischemia, hypertension, excessive intraocular pressure, parkinsonian disorders, eating disorders, seasonal affective disorders, panic disorders, urinary incontinence, diuresis, fertility disorders (including the treatment ofinfertiliby by, for example, in vitro fertilization, and use in antifertility), sexual dysfunction, impotence, postnatal depression, mild stressinduced amenorrhoea, and galactorrrhoea, by administering to a mammal in need thereof a therapeutically effective amount of a compound as described herein, or a pharmaceutically acceptable salt thereof. In addition, such disease states include depression, Alzheimer's disease and diabetes.

Active compounds. Active compounds of the present invention are, in general, pentamidine and analogs thereof. Numerous such compounds are known. In the present invention both benzene rings need not be substituted with an amidine group: only a single ring need be substituted with an amidine group (though optionally both rings may be substituted with an amidine group). Such compounds are known or can be synthesized in accordance with known techniques (See, U.S. Patents Nos.

5,723,495; 5,723,288; 5,686,456; 5,668,167; 5,667,975; 5,643,935; 5,639,755; 5,627,184; 5,622,955; 5,606,058; 5,602,172; 5,594,138; 5,578,631;5,521,189; 5,428,051; 5,206,236; 5,202,320; 4,963,589; 4,940,723; 4,933,347; 4,619,942; 4,397,863; and 4,324,794 (applicants intend that the disclosures of the compounds disclosed in these references are incorporated by reference herein intheir entirety; all of these compounds may be provided with a single amidine group as opposed to two amidine groups). Specific examples of such compounds are set forth below.

A first group of active agents useful for carrying out the present invention are bis-benzenes having the formula I:

X-(CH

2 )n-X

(I)

R3 R3 wherein: A and B are each independently selected from the group consisting of H, loweralkyl, oxyalkyl, nitro, amino, aminoalkyl, halo, hydroxy, carboxy, and compounds of formula

RI-N

I

subject to the proviso that at least one of A and B is a compound of formula subject to the proviso that at least one of A and B is a compound of formula Ri and R 2 are each independently selected from the group consisting of H, loweralkyl, oxyalkyl, alkoxyalkyl, cycloalkyl, aryl, hydroxyalkyl, aminoalkyl and alkylaminoalkyl; or two R 1 groups on the same amidine group together represent

(CH

2 wherein m is 2, 3, or 4;

R

2 and R2 are H; X is 0; and n is A second group of bis-benzenes that may be used to carry out the present invention are those having formula II: A N N B NH HN

(II)

R

1 -N RI N Ri-N RI--N

R

2 (i) subject to the proviso that at least one of A and B is a compound of formula

R

1 and R 2 are each independently selected from the group consisting of H, loweralkyl, oxyalkyl, alkoxyalkyl, cycloalkyl, aryl, hydroxyalkyl, aminoalkyl and 7a alkylaminoalkyl; or two R, group on the same amidine group together represent (0H 2 )m-whereifl m is 2, 3, or 4;

R

3 is H, loweralkyl, oxyalkyl, alkoxyalkyl, hydroxyalkyl, cycloalkyl, aryl, aminoalkyl, alkylaminoalkyl or halogen; X is linear or branched, saturated or unsaturated C1 C12 alkyl containing up to 4 double bonds; or X is a heterocyclic aromatic group selected from the group consisting of: R7 R R 8 6

N

Re R 7 6 R 7

S

R

N

R

-X,

N

R

9

R

6

N

I

R9 R R 6

S

wherein

R

6

R

7 and R 8 are each independently selected from the group consisting of H, lower alkyl, halogen, oxyalkyl, oxyaryl, or oxyarylalkyl;

R

9 is hydrogen, lower alkyl, hydroxy, aminoalkyl, or alkylaminoalkyl; or a pharmaceutically acceptable salt thereof A third group of compounds useful for carrying out the present invention are those having formula III: (C 2n11 C 2n wherein A and B are each independently selected from the group consisting of H, lower alkyl, oxyalkyl, nitro, amino, aminoalkyl, halo, hydroxy, carboxy, and compounds of formula

R-N

I- (i)

R,N

subject to the proviso that at least one of A and B is a compound of formula

R

1 and R 2 are each independently selected from the group consisting of H, lower alkyl, oxyalkyl, alkoxyalkyl, cycloalkyl, aryl, hydroxyalkyl, aminoalkyl and alkylaminoalkyl; or two R 1 groups on the same compound of formula together represent -(CH2)m- wherein m is 2, 3 or 4; or two R 1 groups on the same compound of formula together represent

R-

wherein R 5 is Ri-N (i)

R--N

R,

I

R2

R

3 is H, lower alkyl, oxyalkyl, alkoxyalkyl, hydroxyalkyl, cycloalkyl, aryl, aminoalkyl, alkylaminoalkyl, or halogen; n is an integer from 0 to 2; and X is CH20 or a heterocyclic aromatic group selected from the group consisting of: R7

N

Re R7 R 6

R

7

S

R

6

SN

S

R N R

N

I

R9

S

N

I

F<9

N

wherein Re, R 7 and R 8 are each independently selected from the group consisting of H, lower alkyl, halogen, oxyalkyl, oxyaryl, or oxyarylalkyl; R9 is hydrogen, lower alkyl, hydroxy, aminoalkyl, or alkylaminoalkyl; or a pharmaceutically acceptable salt thereof.

A fourth group of compounds useful for carrying out the present invention are those having formula IV: A 0 B (IV) wherein -11- A and B are each independently selected from the group consisting of H, loweralkyl, oxyalkyl, nitro, amino, aminoalkyl, halo, hydroxy, carboxy, and substituents of formula

RI-N

R2 subject to the proviso that at least one of A and B is a substituent of formula

R

I and R 2 are each indcpendently selected from the group consisting of H, loweralkyl, oxyalkyl, alkoxyalkyl, cyloalkyl, aryl, hydroxyalkyl, aminoalkyl and alkylaminoalkyl; or two RI group on the same amidine group together represent

(CH

2 wherein m is 2, 3, or 4;

R

3 is H, loweralkyl, oxyalkyl, alkoxyalkyl, hydroxyalkyl, cycloalkyl, aryl, aminoalkyl, alkylaminoalkyl or halogen; or two RI groups on the same amidine group together represent Rs-.

wherein R 5 is

RI-N

R2 and the pharmaceutically acceptable salts thereof.

A fifth group of active compounds that may be used to carry out the present invention are those having formula

B

R3 R j^ r( 12wherein: A and B are each independently selected from the group consisting of H, loweralkyl, oxyalkyl, nitro, amino, aminoalkyl, halo, hydroxy, carboxy, and substituents of formula (ii):

R

5

-N

Rj-N

.R

6 subject to the proviso that at least one of A and B is a substituent of formula (ii); R, and R 2 are each independently selected from the group consisting of H, loweralkyl, aryl, alkylaryl, aminoalkyl, aminoaryl; halogen, oxyalkyl, oxyaryl, or oxyarylalkyl;

R

3 and R 4 are each independently selected from the group consisting of H, loweralkyl, oxyalkyl, alkylaryl, aryl, oxyaryl, aminoalkyl, aminoaryl, or halogen; and each Rs is independently selected from the group consisting of H, loweralkyl, alkoxyalkyl, hydroxyalkyl, aminoalkyl, alkyiaminoalkyl, cycloalkyl, aryl, or alkylaryl or two Rs groups together represent C2 to Clo alkyl, hydroxyalkyl, or alkylene; and

R

6 is H, hydroxy, loweralkyl, alkoxyalkyi, hydroxyalkyl, aminoalkyl, alkylamino, alkylaminoalkyl, cycloalkyl, hydroxycycloalkyl, alkoxycycloalkyl, aryl, or alikylaryl; or a pharmaceutically acceptable salt thereof.

A sixth group of active compounds of the present invention are those having formula VI: A

B

(CH

2 )nX (VI)

R

3 R3 wherein: A and B are each independently selected from the group consisting of H, loweralkyl, oxyalkyl, nitro, amino, aminoalkyl, halo, hydroxy, carboxy, and substituents of formula

RI-N

R

2 subject to the proviso that at least one of A and B is a substituent of formula

R

1 and R 2 are each independently selected from the group consisting of H, loweralkyl, oxyalkyl, alkoxyalkyl, cyloalkyl, aryl, hydroxyalkyl, aminoalkyl and alkylaminoalkyl; or two R, group on the same amidine group together represent

(CH

2 wherein m is 2, 3, or 4;

R

3 is H, loweralkyl, oxyalkyl, alkoxyalkyl, hydroxyalkyl, cycloalkyl, aryl, aminoalkyl, alkylaminoalkyl or halogen; or two RI groups on the same amidine group together represent wherein R 5 is

RI-N

R2 X is O, S or NH; n is an integer from 1 to 8; and the pharmaceutically acceptable salts thereof.

Salts of Active Compounds. The active compounds may be converted to a corresponding acid addition salt, or pharmaceutically acceptable salt, by virtue of the presence of the nitrogen atoms.

14- The conversion is accomplished by treatment with at least a stoichiometric amount of an appropriate acid, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like, and organic acids such as acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, malic acid, malonic acid, succinic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid and the like. Typically, the free base is dissolved in an inert organic solvent such as diethyl ether, ethyl acetate, chloroform, ethanol or methanol and the like, and the acid added in a similar solvent.

o0 The temperature is maintained at 0°-50° C. The resulting salt precipitates spontaneously or may be brought out of solution with a less polar solvent.

The acid addition salts of the active compounds may be converted to the corresponding free bases by treatment with at least a stoichiometric amount of a suitable base such as sodium or potassium hydroxide, potassium carbonate, sodium bicarbonate, ammonia, and the like.

Utility and Testing. The compounds are generally useful as described above.

The compounds may be specifically tested in accordance with known techniques.

In one particular aspect, the invention relates to a method of determining the presence of imidazoline receptors in vitro or in vivo in mammalian tissue, especially human tissue, by contacting the sample suspected of containing such receptors with an active compound of the invention and detecting the presence or absence of binding of such compound to a receptor in the sample. The detecting may be carried out by any suitable means, such as by labelling the active compound with a detectable group such as 3 H, or by competitive displacement of another receptor binding compound that it itself labelled, all in accordance with known techniques. Detecting may be carried out with a library or combinatorial library of compounds of the invention, as described below.

In another particular aspect, the invention provides methods of alleviating disease states as described above.

Potential for high selectivity and high affinity for imidazoline sites is determined in vitro by a modification of the method of Brown et al., Br.

J.Pharmacology, Vol. 99, pp 481 varies as 486 (1990), as described in Example 11 of U.S. Patent No. 5,726,197 to Clark et al.

15 Lowering of intraocular pressure is shown in vivo by the method of Moses, R.

Tr. Am. Acad. Opth, and Otol., January-February 1962: 88-95.

Potential for treatment of cerebral ischemia is shown in vitro by a modification of the method of Gotti et al., Brain Res., Vol 522(2), pp 290-307 (1990).

The antihypertensive activity of the compounds may be determined in conscious spontaneous hypertensive rats prepared with indwelling arterial catheter by the in vivo assay described in Popovic V. and Popovic J. Applied. Physiol., Vol.

pp. 727-728 (1960), or a modification thereof.

Administration. In applying the compounds of this invention to treatment of the above conditions, administration of the active compounds and salts described herein can be via any of the accepted modes of administration, including oral, parenteral and otherwise systemic route of administration. Any pharmaceutically acceptable mode of administration can be used, including solid, semi-solid or liquid dosage forms, such as, for example, tablets, suppositories, pills, capsules, powders, liquids, suspensions, or the like, preferably in unit dosage forms suitable for single administration of precise dosages, or in sustained or controlled release dosage forms for the prolonged administration of the compound at a predetermined rate. The compositions will typically include a conventional pharmaceutical carrier or excipient and an active compound or the pharmaceutically acceptable salts thereof and, in addition, may include other medicinal agents, pharmaceutical agents, carriers, adjuvants, etc.

The amount of active compound administered will of course, be dependent on the subject being treated, the severity of the affliction, the manner of administration and the judgment of the prescribing physician. However, an effective dose for oral, parenteral and otherwise systemic routes of administration is in the range of 0.01-20 mg/kg/day, preferably 0.1-10 mg/kg/day. For an average 70 kg human, this would amount to 0.7-1400 mg per day, or preferably 7-700 mg/day.

For solid compositions, conventional non-toxic solid carriers include, for example, pharmaceutical grades of mannitol, lactose, cellulose, cellulose derivatives, sodium crosscarmellose, starch, magnesium stearate, sodium saccharin, talcum, glucose, sucrose, magnesium carbonate, and the like may be used. The active compound as defined above may be formulated as suppositories using, for example, polyalkylene glycols, acetylated triglycerides and the like, as the carrier. Liquid pharmaceutically administrable compositions can, for example, be prepared by dissolving, dispersing, etc. an active compound as defined above and optional pharmaceutical adjuvants in a carrier, such as, for example, water, saline, aqueous dextrose, glycerol, ethanol, and the like, to thereby form a solution or suspension. If desired, the pharmaceutical composition to be administered may also contain minor amounts of nontoxic auxiliary substances such as wetting or emulsifying agents, pH buffering agents and the like, for example, sodium acetate, sorbitan monolaurate, triethanolamine sodium acetate, sorbitan monolaurate, triethanolamine oleate, etc.

Actua! methods of prepai:g such dosage forms are known, or will be apparent, to those skilled in this art; for example, see Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, Pa., 15th Edition, 1975. The composition or formulation to be administered will, in any event, contain a quantity of the active compound(s) in an amount effective to alleviate the symptoms of the subject being treated.

Dosage forms or compositions containing active compounds or their salts) in the range of 0.25 to 95% with the balance made up from non-toxic carrier may be prepared.

For oral administration, a pharmaceutically acceptable non-toxic composition is formed by the incorporation of any of the normally employed excipients, such as, for example pharmaceutical grades of mannitol, lactose, cellulose, cellulose derivatives, sodium crosscarmellose, starch, magnesium stearate, sodium saccharin, talcum, glucose, sucrose, magnesium, carbonate, and the like. Such compositions take the form of solutions, suspensions, tablets, pills, capsules, powders, sustained release formulations and the like. Such compositions may contain 1%-95% active ingredient, more preferably 2-50%, most preferably 5-8%.

Parenteral administration is generally characterized by injection, either subcutaneously, intramuscularly or intravenously. Injectables can be prepared in conventional forms, either as liquid solutions or suspensions, solid forms suitable for solution or suspension in liquid prior to injection, or as emulsions. Suitable excipients are, for example, water, saline, dextrose, glycerol, ethanol or the like. In addition, if desired, the pharmaceutical compositions to be administered may also contain minor amounts of non-toxic auxiliary substances such as wetting or emulsifying agents, pH 17buffering agents and the like, such as for example, sodium acetate, sorbitan monolaurate, triethanolamine oleate, triethanolamine sodium acetate, etc.

A more recently devised approach for parenteral administration employs the implantation of a slow-release or sustained-release system, such that a constant level of dosage is maintained. See, U.S. Pat. No. 3,710,795.

The percentage of active compound contained in such parental compositions is highly dependent on the specific nature thereof, as well as the activity of the compound and the needs of the subject. However, percentages of active ingredient of 0.1% to 10% in solution are employable, and will be higher if the composition is a solid which will be subsequently diluted to the above percentages. Preferably the composition will comprise 0.2-2% of the active agent in solution.

In applying the compounds of the invention to treatment of diseases or disorders of the eye which are associated with an abnormally high intraocular pressure, administration may be achieved by any pharmaceutically acceptable mode of administration which provides adequate local concentrations to provide the desired response. These include direct administration to the eye via drops and controlled release inserts or implants, as well as systemic administration as previously described.

Drops and solutions applied directly to the eye are typically sterilized aqueous solutions containing 0.1% to 10%, most preferably 0.5% to 1% of the active ingredient, along with suitable buffer, stabilizer, and preservative. The total concentration of solutes should be such that, if possible, the resulting solution is isotonic with the lacrimal fluid (though this is not absolutely necessary) and has an equivalent pH in the range of pH 6-8. Typical preservatives are phenyl mercuric acetate, thimerosal, chlorobutanol, and benzalkonium chloride. Typical buffer systems and salts are based on, for example, citrate, borate or phosphate; suitable stabilizers include glycerin and polysorbate 80. The aqueous solutions are formulated simply by dissolving the solutes in a suitable quantity of water, adjusting the pH to about 6.8making a final volume adjustment with additional water, and sterilizing the preparation using methods known to those in the art.

The dosage level of the resulting composition will, of course, depend on the concentration of the drops, the condition of the subject and the individual magnitude of responses to treatment. However, a typical ocular composition could be -18administered at the rate of about 2-10 drops per day per eye of a 0.5% solution of active ingredient.

The compositions of the present invention may also be formulated for administration in any convenient way by analogy with other topical compositions adapted for use in mammals. These compositions may be presented for use in any conventional manner with the aid of any of a wide variety of pharmaceutical carriers or vehicles. For such topical administration, a pharmaceutically acceptable non-toxic formulation can take the form of semisolid, liquid, or solid, such as, for example, gels, creams, lotions, cslutions, suspensions, ointments, powders, or the like. As an example, the active components may be formulated into a gel using ethanol, propylene glycol, propylene carbonate, polyethylene glycols, diisopropyl adipate, glycerol, water, etc., with appropriate gelling agents, such as Carbomers, Klucels, etc.

If desired, the formulation may also contain minor amounts of non-toxic auxiliary substances such as preservatives, antioxidants, pH buffering agents, surface active agents, and the like.

Actual methods of preparing such dosage forms are known, or will be apparent, to those skilled in the art; for example, see, Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, Pa., 16th Edition, 1980.

Combinatorial chemistry. Numerous methods of synthesizing combinatorial libraries and (where desired) applying such molecules on solid supports (where the molecules may be either covalently or non-covalently bound, linked or attached to the solid support) are known, and such libraries can be made and used in accordance with procedures known to those skilled in the art. See, J. Baldwin and I. Henderson, Recent Advances in the Generation of Small-Molecule Combinatorial Libraries: Encoded Split Synthesis and Solid-Phase Synthetic Methodology, Med. Res. Reviews 16, 391-405 (1996); see also U.S. Patent No. 5,565,324 to Still et al., U.S. Patent No.

5,284,514 to Ellman et al., U.S. Patent No. 5,445,934 to Fodor et al. (the disclosures of all United States patents cited herein are to be incorporated herein by reference in their entirety). Libraries of active compounds as described herein a group of compounds that have the general definition of active compounds as described herein, but differ from one another in specific structural features, patterns of substitution(s), etc.), may be screened in accordance with known techniques to identify imidazoline receptor binding compounds with specific desired characteristics.

-19- The present invention is explained in greater detail in the following nonlimiting examples.

EXAMPLE 1 1. Materials and methods Membrane preparation. Membranes were prepared in accordance with known techniques Zonnenchein et al., Eur. J. Pharmacol. 190, 203 215 (1990)). Briefly, male Sprague-Dawley rats (Hilltop, Scottsdale, PA, USA) weighing approximately 250 300 g were decapitated, their livers rapidly removed and immediately placed into 0.25 M cold sucrose solution. The livers were weighed and minced with scissors before being homogenized at 4 0 C in solution A (0.25 M sucrose containing 0.1 mM phenylmethyl sulfonyl fluoride 1:1 Ov/v) for 1 min using a Biospec homogenizer (Bartlesville. OK, USA). The homogenate was centrifuged at 600 x g for 10 min at C. The resulting pellet was washed with solution A, discarded and the supernatant fractions were combined and centrifuged at 12,000 x g for 30 min. This pellet was washed then resuspended in cold buffer B (50 mM Tris/HCl pH Using a Bio- Rad (Hercules, CA, USA) assay kit, the protein concentration was determined using bovine serum albumin as the standard according to the assay of Bradford et al.

(Bradford, 1976. Anal. Biochem. 72, 248 254.). The preparation was stored at -80 OC at a concentration of 20 mg protein /ml.

Saturation binding studies. Binding studies were performed in triplicate in 1 ml buffer B containing 3 H]idazoxan (0.1 50 nM). The addition of rat liver membranes at a final concentration of 500 pg protein /ml initiated binding, and the samples were incubated to equilibrium at 22 OC for 45 min. Non-specific binding was defined as the amount of 3 H]idazoxan bound in the presence of 10 j.M cirazoline.

Competition binding studies. Competition experiments were performed in 1 ml buffer B containing 5 nM 3 H]idazoxan. Increasing concentrations of compound were added (1 nM 10 mM) and binding was initiated by the addition of rat liver membranes at a final concentration of 500 pg protein /ml. In all experiments, bound and free radioligand were separated by vacuum filtration through Whatman GF/C filters and washed with 2 x 10 ml cold buffer B. Radioactivity on filters was determined by liquid scintillation spectroscopy using a Packard Tri-Carb 2100TR.

Samples were run in triplicate and the results were derived from at least three experiments with the exception of Compounds 4 and 8 which were run twice due to limited quantities.

Anti-Pneumocystis carinii activity. Induction, treatment and evaluation of P.

carinii pneumonia in the rat was carried out according to methods described previously (Jones, S.K. et al., Antimicrob. Agents Chemother. 34, 1026 1030 (1990)). To summarize the scoring procedure, the mean histologic scores were determined by two examiners using a blinded protocol. P. carinii cysts were counted iithe stained lung sections and assigned the following values; 0.5 less than 10 cysts counted per two fully examined sections; 1 scattered cysts with less than 10% of the lung tissue involved; 2 scattered cysts with limited intense focal involvement and to 25% of lung tissue involved; 3 scattered cysts with numerous intense areas of focal involvement and 26 to 50% of lung tissue involved; 4 cysts found throughout the tissue with numerous intense focal areas of involvement having greater than of lung tissue involved.

Chemical synthesis. The procedure for the synthesis of 4methoxybenzamidine was adapted from Tidwell et al. Med. Chem. 33, 1252 1257 (1990)). The procedure was altered in the following way; a solution of 1,4dioxane was cooled to -10 OC and presaturated with hydrogen chloride (HCI) gas. 4- Methoxybenzonitrile and anhydrous ethanol were added and the reaction mixture was saturated with HCI gas. The reaction flask was sealed and stirred at ambient temperature for twelve days. The imidate was collected and dried under high vacuum for 20 min and then dissolved in ethanolic ammonia at -10 The reaction mixture was stirred at room temperature overnight. The purification of 4methoxybenzamidine was followed according to the published procedure. All other compounds were synthesized as previously described Berger et al., J.

Pharmacol.Exp. Ther. 256, 883 889 (1991); Tidwell et al., supra (1990)). Final compounds were analyzed using high-performance liquid chromatography, proton nuclear magnetic resonance spectroscopy and elemental analysis.

Drugs. 3 H]Idazoxan (specific activity 45 Ci/mmol) was purchased from Amersham (Arlington Heights, IL, USA), cirazoline from Research Biochemicals International (Natick, MA, USA), phenylmethyl sulfonyl fluoride from Boehringer Mannheim (Indianapolis, IN, USA), sucrose from Fisher (Fair Lawn, NJ, USA) and Tris/HCI from Schwartz/Mann Biotech (Cleveland, OH, USA). Benzamidine, yohimbine, phentolamine, epinephrine and bovine serum albumin were purchased from Sigma (St. Louis, MO, USA). 1,4-Dioxane and 4 -methoxybenzonitrile were purchased from Aldrich (Milwaukee, WI, USA), and HCI gas and ammonia from Matheson (Secaucus, NJ, USA). All other compounds were synthesized in our laboratory. Buffers were prepared using deionized water from Dracor Water Systems (Kensington, MD, USA).

Data analysis. Binding analyses, statistics and graphics were performed using Prism 2.01 (Graphpad Software, Inc., San Diego, CA, USA). Binding parameters for inhibitors are given as mean S.D. except Compounds 4 and 8 which are expressed as mean 95% confidence intervals due to limited supply of compound. The calculated log of the octanol:water partition coefficient (ClogP) was calculated using MedChem Software 3.5 (Daylight Chemical Information Systems, New Orleans, LA,

USA).

2. Results H]Idazoxan saturation binding in rat liver membranes. Rat liver membranes, prepared according to the method ofZonnenschein et al. (supra) were reported to be rich in imidazoline 12 sites while excluding a2-adrenoceptors. Saturation binding analyses reported a Bma of 438 30 fmol/mg protein with non-specific binding representing 10 15% of total binding, which conforms with the results reported by Zonnenschein et al. (1990). 3 H]Idazoxan bound the rat liver membranes with a Kd 14 2 nM for saturable binding. To confirm that a2-adrenoceptors were absent from the preparation, displacement experiments were performed using yohimbine, a selective a,-adrenoceptor antagonist; epinephrine, a selective a 2 adrenoceptor agonist; phentolamine, an a,-adrenoceptor antagonist; and cirazoline, an a 2 -adrenoceptor antagonist as competitive inhibitors of 3 H]idazoxan binding.

Figure 1 shows that selective u 2 -adrenoceptor ligands did not compete for radioligand binding, whereas phentolamine displaced PH] idazoxan binding with low affinity (Ki >5000), and cirazoline with high affinity (Ki 3.0 0.4 nM).

Competition binding studies. A competitive inhibition curve using pentamidine as the inhibitor of [H]idazoxan binding was generated (data not shown).

-22- The Ki of 1.4 0.22 nM demonstrates that pentamidine binds to imidazoline I, sites with very high affinity, comparable to that of the most potent ligands. Several interesting structure/activity relationships were observed from the data in Table 1.

Taking pentamidine (Compound 1) as the lead compound, no change in affinity was observed when the central alkyl link was increased to six carbons (Compound 2) or the amidine groups replaced with imidazoline moieties (Compound Moderate reductions in affinity (10 100 nM) were observed when the alkyl chain was shortened further (Compounds 4 and 7) and a similar reduction was noted when one of.the cations was rcplaced by a nitro group (Corapound Loss of one benzamidine moiety (Compound 9) caused a further reduction in affinity; however, when both the amidine phenoxy moiety and four of the carbons from the alkyl chain were removed from pentamidine (Compound 10), there was no further reduction in affinity when compared to Compound 9.

Interestingly, methoxy substitution meta to the cation (Compounds 5 and 6) produced varied results depending on the nature of the cationic moiety. When compared to the unsubstituted analog (Compound methoxy substitution (Compound 6) resulted in over a two log decrease in imidazoline receptor binding.

However, methoxy substitution of the imidazoline analog (Compound 5) resulted in a compound with similar affinity to Compound 7. The low affinity of Compound 6 may be a result of its very low ClogP. Likewise, a comparison of Compounds 10 12 shows that Compound 11 with the lowest ClogP of all the compounds evaluated also had the lowest affinity. The low affinity of Compound 11 may be due to the existence of contributing resonance structures to form a zwitterion. The basicity and therefore the binding property of the amidines are greatly influenced by the presence of different substituents on the benzamidine ring. Both benzamidine (Compound 12) and 4 -methoxybenzamidine (Compound 10) display resonance stabilization of the positive charge throughout the molecule; however, the phenoxide ion of 4hydroxybenzamidine neutralizes the positive charge, therefore making it unavailable for receptor binding. This molecule lends support to the theory that a strong cation is an important factor in receptor binding.

ClogP. ClogP increases as compound lipophilicity increases. Table 1 shows that Compound 3 is the most lipophilic (ClogP 6.0) and Compound 11 is the least lipophilic (ClogP 0.08) of the compounds tested. There is no significant correlation -23for the group as a whole between Ki and ClogP; Pearson correlation coefficient r 0.38 P 0.22. However, it is interesting to note that the ClogP and Ki values for Compounds 5 and 6 differ considerably. The low affinity of Compound 6 (K 3500 nM) may be attributed to its hydrophilic nature, denoted by the very low ClogP Similarly, when Compounds 10 12 are compared, the derivative with the lowest ClogP (Compound 11) shows weakest affinity for the imidazoline I2 site (Ki 30,000 nM, ClogP 0.08).

24 Table 1. Affinity for 12 binding sites, anti-P.carinii activity and ClogP values for pentamidine (Compound 1) and various analogs.

Structure

K

1 (nM Hl Co Slope gP HN

NH

0-CZ501.4* -0.8 2.3 H1-

NH

2 0 2 2 HN NH

H

2 N NrH 2 N

N

HN

H N 4 N NH 6 0 HN -NH HOCH,

H

3 CO HN

NH

7 HN d NH 3.2 0.36 4.3L, 1.09 23.4 6.8 50. 1 1.06 -1.2 2.8 -0.8 -0.8 1.8 -0.9 -3500.. -0.9 .09 0-(CH 2 )5sNO

HN

\I H N

HN

27.2 7.63 62.0* 319* 39.5 228± 36.5 -30,00 -0.8 1.3 -0.8 3.7 -0.8 .8 -0.8 0 .7 12 580 -0.8 0.6 .61.3 'Mean Histologic Score (comnpared to saline control; mean~ histologic score saline =3.8 out of a possible ND not done. Binding in the presence of 5 nM 3 1-1idazoxan (Kd 14 2nM). 438 30 frno~lmg protein. K values are the result of at least three experiments expressed as mean Ki SD except compounds 4 and 8, which were derived from two experiments due to limited quantities of compound. These results are stated as mean Ki 95% confidence intervals.

Anti-Pneumocystis carinii activity. Pentamidine exhibits anti-P. carinii activity with a mean histologic score of 1.3. This is a moderately effective antimicrobial agent, having activity against a variety of organisms Blagburn et al., Antimicrob.

Agents Chemother. 35, 1520 1523 (1991); C. Bell et al., Antimicrob. Agents Chemother. 35, 1099 1107 (1991); D. Lindsay et al., Antimicrob. Agents Chemother. 35, 1914-1916 (1991)). Five of the compounds have anti-P. carinii activity greater than pentamidine (Compounds while 2 of the. compounds have no activity at all (Compounds 9,11). Compounds 5 and 6 are equally effective anti-P.carinii agents, yet their respective affinities for the imidazoline 12 site differ fold. These compounds provide an example that affinity for the imidazoline 12 site does not correlate to anti-P. carinii activity; Pearson correlation coefficient r 0.60 P 0.09.

EXAMPLE 2 AND 3 The compounds set forth in Table 2 and Table 3 below, which are either known and can be synthesized in accordance with known techniques as described above or variations thereof which will be apparent to those skilled in the art, are also illustrative of the present invention.

-26- TABLE 2: IMIDAZO.LNE RECEPTOR BINDING COMPOUNDS Compound Structure Ki (nM) 001 KAO 011

SAFIM

BABB

BIBB

OB 205 DB 183 003 KAD 111 DAP 092 DIB

HH

N A d NH 2 CN

&K

H H, 158.0 7,512 44,780 346,700 65,640 44,700 18,090 653 .11

H

2 N

NH

2 NH

N

HN 0NHI DB 60 N) 630 H H Ci384 N N-H 5,503 HN NH DB 244 HN oNH675 N

NH

DB26 103N 0 NH 3785 HN

NH

TABLE 3: IMIDAZOLINE RECEPTOR BINDING COMPOUNDS Compound Structue Ki(nM)

H

2 N 2 H 2 MC 96 HJ- H- NH 1.4

H

2

N

FS 44

-CH

2 0- 2.8 FS 104 O CH 2 0

NH

2

I-.

-28 FS 113 HN CH2OCH 3 2.2

CH

2 MC9c

NH

FS 117 CH NH, 32.7

NH

The foregoing is illustrative of the present invention, and is not to be construed as limiting thereof. The invention is defined by the following claims, with equivalents of the claims to be included therein.

Claims

2. The method of Claim 1, wherein the bis-benzene compound comprises a cl compound of formula A B X-(CH 2 (I) R 3 R 3 wherein: A and B are each independently selected from the group consisting of H, loweralkyl, oxyalkyl, nitro, amino, aminoalkyl, halo, hydroxy, carboxy, and compounds of formula R-N (i) R-N R2 subject to the proviso that at least one of A and B is a compound of formula R 1 and R 2 are each independently selected from the group consisting of H, loweralkyl, oxyalkyl, alkoxyalkyl, cycloalkyl, aryl, hydroxyalkyl, aminoalkyl, and alkylaminoalkyl; or two R 1 groups on the same compound of formula together represent -(CH 2 wherein m is 2, 3, or 4; R 3 is H, loweralkyl, oxyalkyl, alkoxyalkyl, hydroxyalkyl, cycloalkyl, aryl, aminoalkyl, alkylaminoalkyl, or halogen; n is an integer from 2 to 6; and XisO, NH, orS; or a pharmaceutically acceptable salt thereof. O 3. The method of Claim 2 wherein the bis-benzene compound comprises a compound selected from the group consisting of: 0 HN ,NH H 0-CH2*nO N ND. H H CH, H H O-CH, HC-O HN. .NH -4CH 2 O H 2 N NH 2 O-CH 3 HC-O and NH HN H 2 N-b /e NH 2 0--CH O wherein n is an integer from 2 to 6; or a pharmaceutically acceptable salt thereof.
4. The method of Claim 1, wherein the bis-benzene compound comprises a compound of formula (II): A B N N T- NH NH wherein: A and B are each independently selected from the group consisting of H, loweralkyl, oxyalkyl, nitro, amino, aminoalkyl, halo, hydroxy, carboxy, and compounds of formula SR-N R-N C(i) O R 2 subject to the proviso that at least one of A and B is a compound of formula R 1 and R 2 are each independently selected from the group consisting of H, c1 loweralkyl, oxyalkyl, alkoxyalkyl, cycloalkyl, aryl, hydroxyalkyl, aminoalkyl, and CN 5 alkylaminoalkyl; O or two R 1 groups on the same compound of formula together represent -(CH 2 wherein m is 2, 3, or 4; 0 X is a linear or branched, saturated or unsaturated C 1 -C1 2 alkyl including up to 4 double bonds; or X is a heterocyclic aromatic group selected from the group consisting of: R7 R 7 R R R N R 8 6 6 6'N N N N R 6 R R 6 R R 6 R 7 O S N I R, Rg wherein R 6 R 7 and Re are each independently selected from the group consisting of H, loweralkyl, halogen, oxyalkyl, oxyaryl, or oxyarylalkyl; Rg is hydrogen, loweralkyl, hydroxy, aminoalkyl, or alkylaminoalkyl; or a pharmaceutically acceptable salt thereof. 0 5. The method of Claim 4, wherein the bis-benzene compound comprises a c compound selected from the group consisting of: H H NH, N N N H H and NH H N HN NH NH HN H N N N wherein n is an integer from 1 to 12; or a pharmaceutically acceptable salt thereof.
6. The method of Claim 1, wherein the bis-benzene compound comprises a compound of formula (III): R 3 R 3 S (CH 2 )n (CH 2 )n (111) A B wherein: A and B are each independently selected from the group consisting of H, loweralkyl, oxyalkyl, nitro, amino, aminoalkyl, halo, hydroxy, carboxy, and compounds of formula R-N (i) R-N R, subject to the proviso that at least one of A and B is a compound of formula ID O R 1 and R 2 are each independently selected from the group consisting of H, loweralkyl, oxyalkyl, alkoxyalkyl, cycloalkyl, aryl, hydroxyalkyl, aminoalkyl and o alkylaminoalkyl; Sor two R 1 groups on the same compound of formula together represent -(CH 2 wherein m is 2, 3, or 4; or two R 1 groups on the same compound of formula together represent R 0 wherein R 5 is (N R-N (i) R-N I R2 R 3 is H, loweralkyl, oxyalkyl, alkoxyalkyl, hydroxyalkyl, cycloalkyl, aryl, aminoalkyl, alkylaminoalkyl, or halogen; n is an integer from 0 to 2; and X is CH 2 0 or a heterocyclic aromatic group selected from the group consisting of: R 7 R, R R R N R 8 6 1 N I N N N R 6 R R 6 R R 6 R O S N R, R6 R R6 N N N 0 S N R9 wherein: R 6 R 7 and R 8 are each independently selected from the group consisting of H, loweralkyl, halogen, oxyalkyl, oxyaryl, or oxyarylalkyl; R 9 is hydrogen, loweralkyl, hydroxy, aminoalkyl, or alkylaminoalkyl; or a pharmaceutically acceptable salt thereof.
7. The method of Claim 6 wherein the bis-benzene compound comprises a compound selected from the group consisting of: N ND H H H3C\ CH3 HC-NH l N C H H 3 C 0N HN-CH H NC NH 3 HN NH HN NH and or a pharmaceutically acceptable salt thereof.
8. The method of Claim 1, wherein the bis-benzene compound comprises a compound of formula (IV): O A B A B A (IV) 0 N 2R3 wherein: c A and B are each independently selected from the group consisting of H, c loweralkyl, oxyalkyl, nitro, amino, aminoalkyl, halo, hydroxy, carboxy, and O 0 5 compounds of formula (N RN R N R, subject to the proviso that at least one of A and B is a compound of formula R 1 and R 2 are each independently selected from the group consisting of H, loweralkyl, oxyalkyl, alkoxyalkyl, cycloalkyl, aryl, hydroxyalkyl, aminoalkyl, and alkylaminoalkyl; or two R 1 groups on the same compound of formula together represent -(CH 2 wherein m is 2, 3, or 4; or two R 1 groups on the same compound of formula together represent R- wherein R 5 is RTN (i) RTN R 2 ;and R 3 is H, loweralkyl, oxyalkyl, alkoxyalkyl, hydroxyalkyl, cycloalkyl, aryl, aminoalkyl, alkylaminoalkyl, or halogen; or a pharmaceutically acceptable salt thereof.
9. The method of Claim 8 wherein the bis-benzene compound comprises a compound selected from the group consisting of: IND NH 2 H 2 N SHN NH H and r s H 2 N -NH2 HN N NH H or a pharmaceutically acceptable salt thereof.
10. The method of Claim 1, wherein the bis-benzene compound comprises a compound of formula R4 B R3 R, A (V) wherein: A and B are each independently selected from the group consisting of H, loweralkyl, oxyalkyl, nitro, amino, aminoalkyl, halo, hydroxy, carboxy, and compounds of formula (ii): RTN (ii) I Re subject to the proviso that at least one of A and B is a compound of formula (ii); R 1 and R 2 are each independently selected from the group consisting of H, loweralkyl, aryl, alkylaryl, aminoaryl, halogen, oxyalkyl, oxyaryl, or oxyarylalkyl; R 3 and R 4 are each independently selected from the group consisting of H, loweralkyl, oxyalkyl, alkylaryl, aryl, oxyaryl, aminoalkyl, aminoaryl, or halogen; ID O each R 5 is independently selected from the group consisting of H, C loweralkyl, alkoxyalkyl, hydroxyalkyl, aminoalkyl, alkylaminoalkyl, cycloalkyl, aryl, O or alkylaryl; or two R 5 groups together represent C2 to C10 alkyl, hydroxyalkyl, or alkylene; and R 6 is H, hydroxy, loweralkyl, alkoxyalkyl, hydroxyalkyl, aminoalkyl, alkylamino, alkylaminoalkyl, cycloalkyl, hydroxycycloalkyl, alkoxycycloalkyl, aryl, Sand alkylaryl; or a pharmaceutically acceptable salt thereof. c 10 11. The method of Claim 1, wherein the bis-benzene compound comprises a compound of formula (VI): A B (CH 2 (VI) R 3 R 3 wherein: A and B are each independently selected from the group consisting of H, loweralkyl, oxyalkyl, nitro, amino, aminoalkyl, halo, hydroxy, carboxy, and compounds of formula R-N (i) R-N I R, subject to the proviso that at least one of A and B is a compound of formula R 1 and R 2 are each independently selected from the group consisting of H, loweralkyl, oxyalkyl, alkoxyalkyl, cycloalkyl, aryl, hydroxyalkyl, aminoalkyl, and alkylaminoalkyl; or two R 1 groups on the same compound of formula together represent -(CH 2 wherein m is 2, 3, or 4; R 3 is H, loweralkyl, oxyalkyl, alkoxyalkyl, hydroxyalkyl, cycloalkyl, aryl, aminoalkyl, alkylaminoalkyl, or halogen; or two R 1 groups on the same compound of formula together represent 38 C) O wherein R 5 is (i) (R-N SR, SX is O, S, or NH; 5 n is an integer from 1 to 8; O Sor a pharmaceutically acceptable salt thereof.
12. The method of Claim 11, wherein the bis-benzene compound comprises a compound selected from the group consisting of: HN O NH H 2 N NH 2 HN S/ CH -O CH-O HN H 2 N Q& Ci-o- O HN H 2 N NH NH 2 HN or a pharmaceutically acceptable salt thereof.
13. The method of Claim 1 wherein the bis-benzene compound comprises a bis-benzamidine.
14. The method of Claim 1 wherein the bis-benzene compound comprises a compound having the following structure: or a pharmaceutically acceptable salt thereof.
15. The method of Claim 1, wherein the subject is afflicted with Alzheimer's disease.
16. The method of Claim 1, wherein the subject is at risk of developing Alzheimer's disease, the treatment is a prophylactic treatment, and the bis- benzene compound is administered in a prophylactically effective amount.
17. A method for treating diabetes in a subject in need of treatment thereof, the method including administering to the subject a therapeutic amount of a bis- benzene compound, wherein said bis-benzene compound comprises at least one amidine group, or a pharmaceutically acceptable salt thereof.
18. The method of Claim 17, wherein the bis-benzene compound comprises a compound of formula (CH2)n-X. wherein: A and B are each independently selected from the group consisting of H, loweralkyl, oxyalkyl, nitro, amino, aminoalkyl, halo, hydroxy, carboxy, and compounds of formula R-N R-N R, IO O subject to the proviso that at least one of A and B is a compound of Sformula O R 1 and R 2 are each independently selected from the group consisting of H, loweralkyl, oxyalkyl, alkoxyalkyl, cycloalkyl, aryl, hydroxyalkyl, aminoalkyl, and alkylaminoalkyl; or two R 1 groups on the same compound of formula together represent -(CH 2 wherein m is 2, 3, or 4; 0 R 3 is H, loweralkyl, oxyalkyl, alkoxyalkyl, hydroxyalkyl, cycloalkyl, aryl, aminoalkyl, alkylaminoalkyl, or halogen; n is an integer from 2 to 6; and X is O, NH, or S; or a pharmaceutically acceptable salt thereof.
19. The method of Claim 18 wherein the bis-benzene compound comprises a compound selected from the group consisting of: HN NH H 2 N 0CHO NH 2 N/ O CH 2 -O ND N O-CH, HC-O HN NH O-CH 3 H 3 C-O and NH HN H 2 N-b NH 2 wherein n is an integer from 2 to 6; or a pharmaceutically acceptable salt thereof. The method of Claim 17, wherein the bis-benzene compound comprises a compound of formula (II): ID A B C N N 0 N N H H c wherein: A and B are each independently selected from the group consisting of H, loweralkyl, oxyalkyl, nitro, amino, aminoalkyl, halo, hydroxy, carboxy, and 0 5 compounds of formula C[i RTN 0 RTN R, subject to the proviso that at least one of A and B is a compound of formula R 1 and R 2 are each independently selected from the group consisting of H, loweralkyl, oxyalkyl, alkoxyalkyl, cycloalkyl, aryl, hydroxyalkyl, aminoalkyl, and alkylaminoalkyl; or two R 1 groups on the same compound of formula together represent -(CH 2 wherein m is 2, 3, or 4; X is a linear or branched, saturated or unsaturated C 1 -C 1 2 alkyl including up to 4 double bonds; or X is a heterocyclic aromatic group selected from the group consisting of: O O C.) 0 0q R, R R 6 N R6 R 7 0 R R N R R7 S R6 N RS N S R 6 N9 wherein R 6 R 7 and R 8 are each independently selected from the group consisting of H, loweralkyl, halogen, oxyalkyl, oxyaryl, or oxyarylalkyl; R 9 is hydrogen, loweralkyl, hydroxy, aminoalkyl, or alkylaminoalkyl; or a pharmaceutically acceptable salt thereof.
21. The method of Claim 20, wherein the bis-benzene compound comprises a compound selected from the group consisting of: NN N N H I H H H and wherein n is an integer from 1 to 12; IND 0 or a pharmaceutically acceptable salt thereof. (N O 22. The method of Claim 17, wherein the bis-benzene compound comprises a O compound of formula (III): R 3 R 3 S (CH 2 X (CH 2 n (111) A B 5 wherein: SA and B are each independently selected from the group consisting of H, loweralkyl, oxyalkyl, nitro, amino, aminoalkyl, halo, hydroxy, carboxy, and compounds of formula R-N (i) R-N R2 subject to the proviso that at least one of A and B is a compound of formula R, and R 2 are each independently selected from the group consisting of H, loweralkyl, oxyalkyl, alkoxyalkyl, cycloalkyl, aryl, hydroxyalkyl, aminoalkyl and alkylaminoalkyl; or two R 1 groups on the same compound of formula together represent (CH2)m- wherein m is 2, 3, or 4; or two R, groups on the same compound of formula together represent R wherein R 5 is RTN (i) R-NI R2 R 3 is H, loweralkyl, oxyalkyl, alkoxyalkyl, hydroxyalkyl, cycloalkyl, aryl, aminoalkyl, alkylaminoalkyl, or halogen; n is an integer from 0 to 2; and X is CH 2 0 or a heterocyclic aromatic group selected from the group consisting of: R7 R6 R7 N S RN R. N I R9 N N RO N 0 R N S wherein: R 6 R 7 and R 8 are each independently selected from the group consisting of H, loweralkyl, halogen, oxyalkyl, oxyaryl, or oxyarylalkyl; R 9 is hydrogen, loweralkyl, hydroxy, aminoalkyl, or alkylaminoalkyl; or a pharmaceutically acceptable salt thereof.
23. The method of Claim 22 wherein the bis-benzene compound comprises a compound selected from the group consisting of: NH 2 I IND SHN NH O H 2 N NH 2 0N 0N o -N N'D H H N 0N HC CH 3 HN NH and N N 00 wh en\ NH HN or a pharmaceutically acceptable salt thereof.
24. The method of Claim 17, wherein the his-benzene compound comprises a compound of formula (IV): B/ (IV N R2 wherein: A and B are each independently selected from the group consisting of H, loweralkyl, oxyalkyl, nitro, amino, aminoalkyl, halo, hydroxy, carboxy, and compounds of formula RTN (i) R-N R subject to the proviso that at least one of A and B is a compound of formula IND O R, and R 2 are each independently selected from the group consisting of H, loweralkyl, oxyalkyl, alkoxyalkyl, cycloalkyl, aryl, hydroxyalkyl, aminoalkyl, and o alkylaminoalkyl; Sor two R 1 groups on the same compound of formula together represent -(CH 2 wherein m is 2, 3, or 4; or two R 1 groups on the same compound of formula together represent R O wherein R 5 is N R-N (i) R-N R2 ;and R 3 is H, loweralkyl, oxyalkyl, alkoxyalkyl, hydroxyalkyl, cycloalkyl, aryl, aminoalkyl, alkylaminoalkyl, or halogen; or a pharmaceutically acceptable salt thereof. The method of Claim 24 wherein the bis-benzene compound comprises a compound selected from the group consisting of: NH 2 H 2 N HN NH N H ;and HN N NH H or a pharmaceutically acceptable salt thereof.
26. The method of Claim 17, wherein the bis-benzene compound comprises a compound of formula IND OR cR, 0 3 2. (V) A 0 0 R2 wherein: O A and B are each independently selected from the group consisting of H, N loweralkyl, oxyalkyl, nitro, amino, aminoalkyl, halo, hydroxy, carboxy, and 0 5 compounds of formula (ii): cN R-N (ii) R-N R 6 subject to the proviso that at least one of A and B is a compound of formula (ii); R 1 and R 2 are each independently selected from the group consisting of H, loweralkyl, aryl, alkylaryl, aminoaryl, halogen, oxyalkyl, oxyaryl, or oxyarylalkyl; R 3 and R 4 are each independently selected from the group consisting of H, loweralkyl, oxyalkyl, alkylaryl, aryl, oxyaryl, aminoalkyl, aminoaryl, or halogen; each R 5 is independently selected from the group consisting of H, loweralkyl, alkoxyalkyl, hydroxyalkyl, aminoalkyl, alkylaminoalkyl, cycloalkyl, aryl, or alkylaryl; or two R 5 groups together represent C2 to C10 alkyl, hydroxyalkyl, or alkylene; and R 6 is H, hydroxy, loweralkyl, alkoxyalkyl, hydroxyalkyl, aminoalkyl, alkylamino, alkylaminoalkyl, cycloalkyl, hydroxycycloalkyl, alkoxycycloalkyl, aryl, and alkylaryl; or a pharmaceutically acceptable salt thereof.
27. The method of Claim 17, wherein the bis-benzene compound comprises a compound of formula (VI): IND SA B (CH 2 )nX- (VI) O R3 R 3 wherein: A and B are each independently selected from the group consisting of H, loweralkyl, oxyalkyl, nitro, amino, aminoalkyl, halo, hydroxy, carboxy, and compounds of formula RTN R_ R, subject to the proviso that at least one of A and B is a compound of formula R 1 and R 2 are each independently selected from the group consisting of H, loweralkyl, oxyalkyl, alkoxyalkyl, cycloalkyl, aryl, hydroxyalkyl, aminoalkyl, and alkylaminoalkyl; or two R 1 groups on the same compound of formula together represent -(CH 2 wherein m is 2, 3, or 4; R 3 is H, loweralkyl, oxyalkyl, alkoxyalkyl, hydroxyalkyl, cycloalkyl, aryl, aminoalkyl, alkylaminoalkyl, or halogen; or two R, groups on the same compound of formula together represent wherein R 5 is RTN RTN R, X is O, S, or NH; n is an integer from 1 to 8; or a pharmaceutically acceptable salt thereof. 49 O 28. The method of Claim 27, wherein the bis-benzene includes a compound c selected from the group consisting of: SHN C /NH o cH-CHO- CH-O 0 5 NH2, SHN- CH2-O /O-CH 3 H 2 N H 2 N =NH H 2 N and -NH 2 HN or a pharmaceutically acceptable salt thereof.
29. The method of Claim 17 wherein the bis-benzene compound comprises a bis-benzamidine. The method of Claim 17 wherein the bis-benzene compound comprises a compound having the following structure: NH or a pharmaceutically acceptable salt thereof.
31. The method of Claim 17, wherein the subject is afflicted with diabetes.
32. The method of Claim 17, wherein the subject is at risk of developing diabetes, the treatment is a prophylactic treatment, and the bis-benzene compound is administered in a prophylactically effective amount.