ZA200004087B

ZA200004087B - Novel therapeutic agents that modulate alpha-1A adrenergic receptors.

Info

Publication number: ZA200004087B
Application number: ZA200004087A
Authority: ZA
Inventors: John H Griffin; Edmund J Moran; G Roger Thomas
Original assignee: Advanced Medicine Inc
Priority date: 1998-06-08
Filing date: 2000-08-10
Publication date: 2001-11-13
Also published as: ZA200004565B; ZA200004084B; ZA200004085B; ZA200003475B; ZA200004561B; ZA200004083B

Description

NOVEL THERAPEUTIC AGENTS THAT MODULATE

ALPH.A-1A ADRENERGIC RECEPTORS

BACKGROUND OF THE INVENTION Field of the Invention

This invention relat-es to novel therapeutic agents which bind to mammalian receptors and modulate the ir activity. More particularly, the invention relates to novel therapeutic agents that bind to and modulate the in vivo activity of alpha-1A adrenergic receptors in marnmals by acting as multi-binding compounds. The therapeutic agents or multi—binding compounds described herein comprise at least two ligands connected by a linker or linkers, wherein said ligands in their monovalent state bind to an d/or are capable of modulating the activity of the alpha- 1A adrenergic receptor. The linking moiety is chosen such that the multi-binding compounds so constructed clemonstrate increased biological activity as compared to individual units of the lig and. The invention also relates to methods of using such compounds, to method s of preparing such compounds and to pharmaceutical compositions containing them.

These multi-binding -compounds are particularly useful in treating mammalian conditions that are mediated by the alpha-1A adrenergic receptors targeted by the ligands, sucha as benign prostatic hyperplasia (“BPH”) and hypertension. Accordingly, this invention also relates to pharmaceutical compositions comprising a pharmaceuti cally acceptable excipient and an effective amount of a multi-binding compound off this invention.

Additionally, the multi-binding <ompounds are useful as affinity resins for affinity chromatography. When so employed, the compounds of the invention may be used as a tool in immunoprecipitatio n. The compounds may be used to identify a receptor in vitro for example in microscopy, electrophoresis and chromatography.

References

The following publications are cited in this application as superscript numbers: 1. J. March, Advanced Organic Chemistry, Ye Edition, Wiley-

Interscience New York 1992); 2. Remington's Pharmaceatical Sciences, Mace Publishing Company,

Philadelphia, PA, 17th ed. (1985); 3. Green, Protective Groups in Organic Synthesis, 2M Edition, John

Wiley & Sons, New Yo rk, New York (1991); 4. Bock, et al., U.S. Paterat 5,807,856, issued September 15, 1998; 5. Tammela, “Benign Prostatic Hyperplasia Practical Treatment

Guidelines,” Drugs & Aging (1997) 10(5):349-366; 6. Wong et al., “Identification of a Dihydropyridine as.a Potent a1 A

Adrenoceptor-Selective Antagonist That Inhibits Phenylephrine-

Induced Contraction of the Human Prostate”, J. Med. Chem. (1998) 41(14):2643-2650; 1. Lee et al., “Clinical Comparison of Selective and Non-selective

Alpha-1A Adrenorecepetor Antagonists in Benign Prostatic

Hyperplasia: Studies ox1 Tamsulosin in a Fixed Dose and Terazosin in Increasing Doses,” 2r. J. Urology (1997) 80:606-611;

8. Chang et al., “Molecular Cloning, Genomic Characterization ard

Expression of Novel Human a1A-Adrenoceptor isoforms,” FEBS

Let. (1998) 422:279-283; 9. Ford et al., “Pharmacological Pleiotropism of the Human

Recombinant Alpha 1A-adrenoceptor: Implications for Alphal- adrenoceptor classification,” Br. J. Pharmacol. (1997) 121(6): 1127-1135; 10. Richard son et al., “Pharmacology of Tamsulosin: Saturation- : Binding Isotherms and Competition Analysis Using Cloned o-

Adrenergic Receptor Subtypes,” The Prostate (1997) 33:55-59; 11. Haynes etal., “B-Adrenoceptor-Mediated Inhibition of a; -

Adrenoceptor-Mediated and Field Stimulation-induced Contractile

Responses in the Prostate of the Guinea Pig,” Br. J. Pharmaco. (1997) 112(6):1067-1074; 12. Hancock et al., “Actions of A-131701, a Novel, Selective

Antagonist for Alpha-1A Compared with Alpha-1B Adrenoceptors on Intrawrethral and Blood Pressure Responses in Conscious Do _gs and a Pharmacodynamic Assessment of In Vivo Prostatic

Selectivity,” J. Pharmacol. Exper. Ther. (1998) 285:628-642; 13. Meyer et al., “Synthesis and Pharmacological Characterization of 3- [2-((3aR2,9bR)-cis-6-Methoxy-2,3,3a,4,5,9b-hexahydro-{1 H}- benz[e]i soindol-2-yl)ethyl]pyrido[3',4':4,5]thieno(3,2-d]pyrimidiine- 2,4(ir1,3H-dione; (A-i131701): A Uroseiective a, Adrenoceptor

Antagonist for the Symptomatic Treatment of Bening Prostatic

Hyperplasia,” J. Med. Chem. (1997) 40(20):3142-3143;

All of the above publications are herein incorporated by reference in their entirety to the same exxent as if each individual publication was specifically ancl individually indicated to be incorporated by reference in its entirety.

State of the Art

A receptor is a biological structure with one or more binding domains that reversibly complexes with one or more ligands, where that complexation has biological consequences. Receptors can exist entirely outside the cell

(extracellular receptors), within the cell membrane (but presenting sections of the receptor to the extracellular milieu and cytosol), or entirely within the cell (intracellular receptors). They may also function independently of a cell (e.g., clot formation). Receptors within the cell mexmbrane allow a cell to communicate with the space outside of its boundaries (i.e., signaling) as well as to function in the transport of molecules and ions into and out of the cell.

A ligand is a binding partner for a specific receptor or family of receptors.

A ligand may be the endogenous ligand for the receptor or alternatively may be a synthetic ligand for the receptor such as a drug, a drug candidate or a pharmacological tool.

The ligands that bind to cellular receptors may be specifically classified as follows: 1) Full agonists - ligands that when bound trigger the maximum activity seen by natural ligands; 2) Partial agonists- ligands that w hen bound trigger sub-maximal activity; 3) Antagonist- ligands that when bound inhibit or prevent the activity arising from a natural ligand binding to the receptor. Antagonists may be of the surmountable class (results in the parallel displacement of the dose-response curve of the agonist to the right in a dose dependent fashion without reducing the maximal response for the agonist) or insurmountable class (results in depression of the maximal response for a given agonist with or without the parallel shift); 4) Inverse antagonist-ligands that when bound decrease the basal activity of the unbound receptor (if any).

There are four fundamental measurable properties that pertain to the interaction of a ligand with its receptor:

1) the affinity of the ligand for the receptor, “which relates to the energetics of the binding; 2) the efficacy of the ligand for the receptor, which relates to the functional downstream activity of the ligand; 3) the kinetics of the ligand for the receptor, which defines the onset of action and the duration of action; and 4) the desensitization of the receptor for the ligand.

With regard to the ligand, it is the combination of these properties that provides the foundation for defining the nature of the= functional response. Thus, an activating ligand (or agonist) has affinity for the reeceptor and downstream efficacy. In contrast, an inhibiting ligand (antagonist) has affinity for the receptor - but no efficacy.

Selectivity defines the ratios of affinities or th e ratios of efficacies of a given ligand compared across two receptors. It is the selectivity of a specific drug . 15 that provides the required biological profile.

Current drugs (ligands) targeting receptors have clinica. shortcomings identified by one or more of low efficacy, low affinit=y, poor safety profile, lack of selectivity or overselectivity for the intended receptor, and suboptimal duration of action and onset of action. Accordingly, it would be beneficial to develop ligands that have improved affinity, efficacy, selectivity, onset of action and duration of action. } ffinity of ligand fi

An increase in ligand affinity to the target receptor may contribute to reducing the dose of ligand required to induce the des ired therapeutic effect. A reduction dn ligand affinity will remove activity and may contribute to the selectivity profile for a ligand.

Efficacy o f ligand at a target receptor (functional effect)

Am increased ligand efficacy at a target receptor can lead to a reduction in the dose resquired to mediate the desired therapeutic effect. This increase in efficacy mmay arise from an improved positive functional response of the ligand or a change from a partial to full agonist profile. Reduced efficacy of a full agonuist to a partial agonist or antagonist may provide clinical benefit by modulating the biological response.

Selectivity~ of ligand compared across receptor subtypes

Am increase in the selectivity of the ligand across receptor subtypes requires that the affinity or efficacy of the ligand at other receptors is reduce=d relative to the desired receptor. A decrease in the selectivity of the ligand may also be desired. .

Onset of Action

Moore rapid onset of action of the ligand to effect a biological response is often preferred.

Duration eof Action

Ar increased duration of action of the ligand to effect a biological rexsponse may be prreferred. For example 3, adrenergic agonists such as albuterol have a relatively short duration of action of approximately 3-4 hours and an increase in duration Of action would simplify the dosing regimen required to administexc this drug (ligand).

Desensitization is best defined as the variety of processes by which the functional imteraction of the receptor with its G-protein are influenced. These processes le=ad ultimately to a reduction in cellular response to th_e activating agonist. Such phenomena are most often observed during prolonged stimulation of the receptor. The two main pathways for receptor desensitizatiorm are reduction in receptor dersity or changes in receptor structure by phosphorylation mechanisms.

Receptor density is altered by receptor sequestration. Th is is a reversible process that is observable within minutes and is a dynamic sorting of receptors with receptosrs being cycled to and from the membrane. On the Other hand, receptor downregulation is generally slower, in the order of hours, and is irreversible, involving destruction of the receptor. Finally, receptor density may be affected Dy an alteration in the rate of synthesis. Alternatively, receptor desensitizati<on may occur through changes in receptor structure, such as receptor phosphoryla tion.

Receptor oligomerization also plays a role in receptor function. The family aipha-. adrenergic receptors (aipha-1A, alpha-i3, alpha-iD and alpha-1L), mediate vari ous actions of the peripheral sympathetic nervous sy stem through the binding of the catecholamines, epinephrine and norepinephrine.

Hum an adrenergic receptors are integral membrane proteins which have been classified into two broad classes, the alpha and the beta adrenergic receptors.

Both types maediate the action of the peripheral sympathetic nervous system upon binding of catecholamines, norepinephrine and epinephrine.

Nore pinephrine is produced by adrenergic nerve endings, while epinephrine is produced by the adrenal medulla. The binding affinity of adrenergic receptors for these compounds forms one basis of the classification.

Alpha receptors bind norepinephrine more strongly thhan epinephrine and much more strongly than the synthetic compound isoproterenol. The binding affinity of these hormones is reversed for the beta receptors. Im. many tissues, the functional responses, such as smooth muscle contraction, induced by alpha receptor activation are opposed to responses induced by beta receptor binding. *

Benign prostatic hyperplasia (“BPH”) is a progressive condition which is characterized by a nodular enlargement of prostatic tx ssue resulting in obstruction of the urethra.® This condition occurs in over 50% of the male population above age 60 and leads to a variety of urological symptoms including increased “frequency in urination, nocturia, a poor urine stream and hesitancy or delay in starting the urine flow.® Further, BPH can lead to lo wer urinary tract infections and benign prostatic enlargement. Currently, a variety of alpha-1 adrenergic receptor antagonists are used clinically to treat BPH which relax the smooth muscle of the prostate.’ However, these agents have been shown to cause significant side effects including dizziness, decreased blood pressure, nasal congestion and impotence, presumably as a result of ®heir lack of selectivity for any one of the receptor subtypes. Examples of these antagonists include prazosin, terazosin, doxazosin and alfuzosin.® It has been demonstrated that the alpha-1A subtype is the predominant receptor mediating smooth muscle contraction in the human prostate. Thus, an alpha-1A selective antagomnist will show better efficacy in the treatment of BPH, with reduced side effects, thhan nonselective alpha-1 antagonists.

Further, because the alpha-1A subtype mediates smooth vascular muscle contraction, it is useful in the treatment of hypertenskon and other related cardiovascular disorders.

Accordingly, novel ligands having desired potency and therapeutic effec t for the alpha-1.A adrenergic receptor would be particularly desirable in order to treat benign preostatic hyperplasia and hypertension in mammalian patients. Suc-h novel ligands vwould preferably achieve the desired potency and therapeutic effesct by modulating one or more of the ligand’s properties as to efficacy, affinity, safety profile, selectivity, duration of action and/or onset of action.

SUMMARY OF THE INVENTION

This invention is directed to general synthetic methods for generating large libraries of diverse multimeric compounds which multimeric compounds are candidates for possessing multibinding properties. The diverse multimeric compound libraries provided by this invention are synthesized by combining a linker or linkers with a ligand or ligands to provide for a library of multimeric compounds wherein the linker and ligand each have complementary functional groups permitting covalent linkage. The library of linkers is preferably selected_ to have diverse pr operties such as valency, linker length, linker geometry and rigidity, hydrophilicity or hydrophobicity, amphiphilicity, acidity, basicity and polarization. T he library of ligands is preferably selected to have diverse attachment poirats on the same .igand, different functionai groups at the same sit = of otherwise the same ligand, and the like.

This inv-ention is also directed to libraries of diverse multimeric compoumnds which multimeric compounds are candidates for possessing multibinding properties. Thesse libraries are prepared via the methods described above and permit the rapic and efficient evaluation of what molecular constraints impart multibinding properties to a ligand or a class of ligands targeting a receptor.

This inv-ention is directed, in part, to novel multi-binding compounds thaat bind alpha-1A adrenergic receptors and consequently these compounds can be u. sed to treat conditions mediated by alpha-1A adrenergic receptors such as benign prostatic hyperplasia and hypertension.

Accordingly, in one of its composition as pects, this invention is directed to a multi-binding compound and salts thereof compprising 2 to 10 ligands, which may be the same or different and which are covalently attached to a linker or linkers which may be the same of different, at least one of said ligands comprising a ligand domain capable of binding to an alpha-1A_ adrenergic receptor. Preferably, at least two and more preferably each of the ligamds comprises a ligand domain capable of binding to an alpha-1A adrenergic receptor.

The multi-binding compounds of this invention are preferably represented by formula I:

L)pX)q I wherein cach L is independently selected from li gands comprising a ligand domain capable of binding to an alpha-1A adrenergic receptor; X is independently a linker; pis an integer of from 2 to 10; gq is an integer of from 1 to 20; and pharmaceutically acceptable salts thereof. Prefemably, g is less than p.

In another of its composition aspects, this invention is directed to a pharmaceutical composition comprising a pharmaceutically acceptable excipient and an effective amount of a multi-binding compeound, or a pharmaceutically acceptable salt thereof, comprising 2 to 10 ligands which may be the same or different and which are covalently attached to a Rinker or linkers which may be the same or different, at least one of said ligands comprising a ligand domain capable of binding to one or more alpha-1A adrenergic receptors.

Preferably, said ligands comprising a ligand domain capable of binding &o one or more alpha-1A adrenergic receptors that modulate benign prostatic hyperplasia in mammals. More preferably, said ligands are selected from the group consisting of terazosin, prazosin, doxasosin, alfuzosin, tamsulosin, RS 100975 (Roche Biosciences), A-131701 (Abbott), 1794-191 (Merck), L757464 (Merck), REC 15-2739 (SB216469, Recordati/SKB), KMD-3213 (Kissei

Pharmaceuticals), and derivatives thereof.

In still another of its composition aspects, this invention is directed to a pharmaceutical composi tion comprising a pharmaceutically acceptable excipiemt and an effective amount o f a multi-binding compound represented by formula I: (L)p(X), I wherein each L is independently selected from ligands comprising a ligand domain capable of binding to an alpha-1A adrenergic receptor; X is a linker; p is an integer of from 2 to 10; gq is an integer of from 1 to 20; and pharmaceutically acceptable salts thereof. Preferably, g is less than p, and more preferably the ligand is selected from the group consisting of terazesin, prazosia, doxascsia, alfuzosin, tamsulosin, RS 100975 (Roche Biosciences), A-131701 (Abbott), L794- 191 (Merck), L757464 (Merck), REC 15-2739 (SB216469, Recordati/SKB),

KMD-3213 (Kissei Pharmaceuticals), and derivatives thereof.

In one of its method aspects, this invention is directed to a method for treating benign prostatic hyperplasia and/or hypertension in a mammal mediated by alpha-1A adrenergic receptors which method comprises administering to saidl mammal an effective amownt of a pharmaceutical composition comprising a pharmaceutically acceptable excipient and a multi-binding compound, or a pharmaceutically acceptable salt thereof, comprising 2 to 10 ligands which may be the same or different and which are covalently attached to a linker or linkers which may be the same or different, at least two of said ligands comprising a ligand domain capable of binding to an alpha-1A adrenergic receptor.

In another of its method aspects, this invention is directed to a method for treating benign prostatic hyperplasia and/or hypertension in a mammal mediated by alpha-1A adrenergic receptors which method comprises administering to said mammal an effective amount of a pharmaceutical composition comprising a pharmaceutically acceptable excipient and a multi-binding compound represented by formula I: (L)p(X)q I wherein each L is independently selected from ligands comprising a ligand domain capable of binding to an alpha-1A adrenergic receptor; X is a linker; p is an integer of from 2 to 10; g is an integer of from 1 to 20 and pharmaceutically acceptable salts thereof.

Preferably, gq is less than p, and more preferably, the ligand is selected from the group consisting of terazosin, prazosin, doxasosin, alfuzosin, tamsulosin,

RS 100975 (Roche Biosciences), A-131701 (Abbott), 1794-191 (Merck), L757464 (Merck), REC 15-2739 (SB216469, R.ecordati/SKB), KMD-3213 (Kissei

Pharmaceuticals), and derivatives ther eof.

Accordingly, in one of its methiod aspects, this invention is directed to a method for identifying multimeric ligand compounds possessing multibinding properties which method comprises:

(a) identifying a ligand or a mixwre of ligands wherein each ligand is capable of binding to an alpha-1A adrene=rgic receptor and contains at least one reactive functionality; (b) identifying a library of lin kers wherein each linker in said library comprises at least two functional groups having complementary reactivity to at least one of the reactive functional group s of the ligand; (©) preparing a multimeric lig and compound library by combining at least two stoichiometric equivalents of th _e ligand or mixture of ligands identified in (a) with the library of linkers identifie<] in (b) under conditions wherein the complementary functional groups react to form a covalent linkage between said linker and at least two of said ligands; amd (d) assaying the multimeric liggand compounds produced in (c) above to identify multimeric ligand compounds posssessing multibinding properties.

In another of its method aspects, &his invention is directed to a method for identifying multimeric ligand compounds possessing multibinding properties which method comprises: (a) identifying a library of ligaands wherein each ligand is capable of tinding °c an alpha-1A adrenergic secept cr and contains at least one reactive functionality; b) identifying a linker or mixzture of linkers wherein each linker comprises at least two functional groups having complementary reactivity to at least one of the reactive functional group s of the ligand, (c) preparing a multimeric lig and compound library by combining at least two stoichiometric equivalents of th_e library of ligands identified in (a) with the linker or mixture of linkers identified in (b) under conditions wherein the complementary functional groups react to form a covalent linkage between said linker and at least two of said ligands; arad d) assaying the multimeric ligand compounds produced in (c) above to identify multimeric ligzand compounds possessing multibinding properties.

The preparatiom of the multimeric ligand compound library is achieved by either the sequential ox concurrent combination of the two or more stoichiometric equivalents of the ligamnds identified in (a) with the linkers identified in (b).

Sequential addition is gpreferred when a mixture of different ligands is employed to ensure heterodimeric ©r multimeric compounds are prepared. Concurrent addition of the ligands is preferred when at least a portion of the multimeric compounds prepared are homomuk timeric compounds.

The assay protocols recited in (d) can be conducted on the multimeric ligand compound libra_ry produced in (c) above, or preferably, each member of the library is isolated by pereparative liquid chromatography mass spectrometry (LCMS).

In one of its cosmposition aspects, this invention is directed to a library of multimeric ligand comapounds which may possess multivalent properties which library is prepared by the method comprising: (a) identify-ing a ligand or a mixtwre of ligands wherein each ligand is capable of binding to &n alpha-1A adrenergic receptor and contains at least one reactive functionality; (b) identify» ing a library of linkers wherein each linker in said library comprises at least two functional groups having complementary reactivity to at least one of the reactiwe functional groups of the ligand; and (©) preparing a multimeric ligand compound library by combining at least two stoichiometr-ic equivalents of the ligand or mixture of ligands identified in (a) with the library of linkers identified in (b) under conditions wherein the complementary functional groups react to form a covalent linkage between said linker and at least two of said ligands.

In another of its composition aspects, this invention is directed to a library of multimeric ligand compounds which may possess multivalent properties which library is prepared by the method comprising: (a) identifying a library of ligands wherein each ligand is capable of binding to an alpha-1A adrenergic receptor and contains at least one reactive functionality; (b) identifying a linker or mixture of linkers wherein each linker comprises at least two functional groups having complementary reactivity to at least one of the reactive functional groups of the ligand; and © preparing a multimeric ligand compound library by combining at least two stoichiometric equ ivalents of the library of ligands identified in (a) with the linker or mixture of linkers identified in (b) under conditions wherein the complementary functional groups react to form a covalent linkage between said linker and at least two of said ligands. in a preferred embodiment, the library of linkers empioyed in either the methods or the library aspects of this invention is selected from the group comprising flexible linkers, rigid linkers, hydrophobic linkers, hydrophilic linkers, linkers of different geometry, acidic linkers, basic linkers, linkers of different polarization and/or polarizability and amphiphilic linkers. For example, in one embodiment, each of the linkers in the linker library may comprise linkers of different chain length and/or having different complementary reactive groups.

Such linker lengths can preferably range from about 2 to 100A.

In another preferred embodiment, the ligand or mixture of ligands is selected to have reactive functionality at different sites on said ligands in order to provide for a range of orientations of said ligand on said multimeric ligand compounds. Such reactive functionality includes, by way of example, carboxylic acids, carboxylic acid halides, carboxyl esters, amines, halides, pseudohalides, isocyanates, vinyl unsaturation, ketones, aldehydes, thiols, alcohols, anhydrides, boronates and precursors thereof. It is understood, of course, that the reactive functionality on the ligand is selected to be complementary to at least one of the reactive groups on the linker so that a covalent linkage can be formed between the linker and the ligand.

In other embodiments, the multimeric ligand compound is homomcric (i.c., each of the ligands is the same ligand having a ligand binding domain capable of binding to an alpha-1A adrenergic receptor, although it may be attached at different points) or heteromeric (i.e. , at least one of the ligands is different from the other ligands).

In addition to the combinatorial methods described herein, this invention provides for an iterative process for rationally evaluating what molecular constraints impart multibinding properties to a class of multimeric compounds or ligands targeting a receptor. Specifically, this method aspect is directed to a method for identifying multimeric ligand compounds possessing multibinding properties which method comprises: (a) preparing a first collection or iteration of multimeric compounds which is prepared by contacting at least two stoichiometric equivalents of the ligand or mixture of ligands which target a receptor with a linker or mixture of linkers wherein said ligand or mixture of ligands comprises at least one reactive functionality and said linker or mixture of linkers comprises at least two functional groups having complementary reactivity to at least one of the reactive functional groups of the ligand wherein said contacting is conducted under conditions wherein the complementary functional groups react to form a covalent linkage between said linker and at least two of said ligands; (b) assaying said first collection or iteration of multimeric compounds to assess which if any of said multimeric compounds possess multibinding = properties; (c) repeating the process of (a) and (b) above until at least one multimeric compound is found to possess multibinding properties; (d) evaluating what molecular constraints imparted multibinding properties to the multimeric compound or compounds found in the first iteration recited in (a)- (c) abave; (e) creating a second collection or iteration of multimeric compounds which elaborates upon the particular molecular constraints imparting multibinding properties to the multimeric compound or compounds found in said first iteration;

H evaluating what molecular constraints imparted enhanced multibinding properties to the multimeric compound or compounds found in the second collection or iteration recited in (e) above; (® optionally repeating steps (e) and (f) to further elaborate upon said molecular constraints.

Preferably, steps (e) and (f) are repeated at least two times, more preferably at least from 2-50 times, even more preferably from at least 3 to 50 times, and still more preferably at least 5-50 times.

DETAILED DESCRIPTION OF THE INVENTION

Ligand (drug) interactions with cellular receptors are controlled by molecular interaction/recognition between the ligand and the receptor. In turn, such interaction can result in modulation or disruption of the biological processes/functions of these receptors and, in some cases, leads to cell death.

Accordingly, when cellular receptors mediate mammalian pathologic conditions,

interactions of the ligand with the cellular receptor can be used to treat these conditions. Of particular interest are tnammalian alpha-1A adrenergic receptors which are known to mediate the contraction and relaxation of the smooth muscles of the prostate gland in mammals. As noted above, this invention is directed, in part, to multi-binding compounds that bind alpha-1A adrenergic receptors.

The “affinity” and “specificity ” of the alpha-1A adrenergic receptor and a ligand thereto are dependent upon the complementarity of molecular binding surfaces and the energetic costs of complexation. “Affinity” is sometimes quantified by the equilibrium constant of complex formation. Specificity relates to the difference in affinity between the same ligand binding to different ligand binding sites on the cellular receptor.

The multi-binding compounds of this invention are capable of acting as multi-binding agents and the surprising activity of these compounds arises at least in part from their ability to bind in a multivalent manner with mammalian alpha- 1A adrenergic receptors. Multivalent binding interactions are characterized by the concurrent interaction of multiple ligands with multiple ligand binding sites on one or more alpha-1A adrenergic receptors. Multivalent interactions differ from collections of individual monovalent interactions by imparting enhanced biological and/or therapeutic effect. Examples of multivalent binding interactions (e. g., 1rivalent) relative to monovalent binding interactions are shown below:

RE ~ — (&

IT RSNTT

Just as multivalent binding can amplify binding affinities, it can also amplify differences in binding affinities, resulting in enhanced binding specificity as well as affinity.

Defigiti ]

Prior to discussing this invention in further detail, the following terms will first be defaned.

The term "alkyl" refers to a monoradical branched or unbranched satzrated hydrocarbo n chain preferably having from 1 to 40 carbon atoms, more preferably 1 10 10 carbon atoms, and even more preferably 1 to 6 carbon atoms. This te=rm is exemplified by groups such as methyl, ethyl, n-propyl. iso-propyl, n-butyl, is o- butyl, n-hexyl, n-decyl, tetradecyl, and the like.

The term "substituted alkyl" refers to an alkyl group as defined above having from | to 5 substituents, and preferably 1 to 3 substituents, selected freom the group consisting of alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, subs tituted aminc, amincacy!, amiroacyioxy, cxyamincacy:, azide, cy-ano, halogen, hyciroxyl, keto, thioketo, carboxyl, carboxylalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkox y, aryl, aryloxy, heteroaryl, heteroaryloxy, heterocyclic, heterocyclooxy, hydroxy- amino, alkoxyamino, nitro, -SO-alkyl, -SO-substituted alkyl, -SO-ary!, -SO- heteroaryl, - SO,-alkyl, -SO,-substituted alkyl, -SO,-aryl and -SO,-heteroaryl.

The t erm "alkylene" refers to a diradical of a branched or unbranched saturated hydrocarbon chain, preferably having from 1 to 40 carbon atoms, more preferably 1 to 10 carbon atoms and even more preferably 1 to 6 carbon atoms . ~

This term is exemplified by groups such as methylene (-CH,-), ethylene

(-CH,CH,-), the propylene isomers (e.g., -CH,CH,C H,- and -CH(CH;)CH,-) and the like.

The term "substituted alkylene" refers to an all<ylene group, as defined above, having from 1 to 5 substituents, and preferably 1 to 3 substituents, selected from the group consisting of alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy, ©xyaminoacyl, azido, cyano, halogen, hydroxyl, keto, thioketo, carboxyl, carboxylalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol, thicalkoxy, substituted thioaikoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, heterocyclic, heterocyclooxy, hydroxyamino, alkoxyamino, nitro, -SO-alkyl, -SO-stabstituted alkyl, -SO-aryl, -

SO-heteroaryl, -SO,-alkyl, -SO,-substituted alkyl, -SO,-aryl and -SO,-heteroaryl.

Additionally, such substituted alkylene groups includes those where 2 substituents on the alkylene group are fused to form one or more cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, ary/1, heterocyclic or heteroaryl groups fused to the alkylene group. Preferably such fused groups contain from 1 to 3 fused ring structures.

The term "alkaryl" refers to the groups -alkylesne-aryl and -substituted alkylene-aryl where alkylene, substituted alkylene and aryl are defined herein.

Such alkaryl groups are exemplified by benzyl, phenethyl and the like.

The term "alkoxy" refers to the groups alkyl-O-, alkenyl-O-, cycloalkyl-O- » cycloalkenyl-O-, and alkynyl-O-, where alkyl, alkenyl, cycloalkyl, cycloalkenyl, and alkynyl are as defined herein. Preferred alkoxy groups are alkyl-O- and include, by way of example, methoxy, ethoxy, n-propoxy, iso- propoxy, n-butoxy, tert-butoxy, sec-butoxy, n-pentox.y, n-hexoxy, 1,2- dimethylbutoxy, and the like.

The term "substituted alkoxy" refers to the groups substitute alkyl-O-, substituted alkenyl-O-, substituted cycloalkyl-O-, substituted cycloal kenyl-O-, and substituted alkynyl-O- where substituted alkyl, substituted alkenyl, s-ubstituted cycloalkyl, substituted cycloalkenyl and substituted alkynyl are as defined herein.

The term "alkylalkoxy" refers to the groups -alkylene-O-alky~1, -alkylene-

O-substituted alkyl, -substituted alkylene-O-alkyl and -substituted alkylene-O- substituted alkyl wherein alkyl, substituted alkyl, alkylene and subst ituted alkylene are as defined herein. Preferred alkylalkoxy groups are alkylene-O—alkyl and include, by way of example, methylenemethoxy (-CH,OCH,), ethykenemethoxy (-

CH,CH,OCHj3), n-propylene-iso-propoxy (-CH,CH,CH,0CH(CH=),), methylene-7—butoxy (-CH,-O-C(CHs)3) and the like.

The term "alkylthioalkoxy" refers to the group -alkylene-S-a1kyl, alkylene-

S-substituted alkyl, substituted alkylene-S-alkyl and substituted alky Iene-S- substituted alkyl wherein alkyl, substituted alkyl, alkylene and subst ituted alkylene are as defined herein. Preferred alkylthioalkoxy groups are alkylene-S-alkyl and include, by way of example, methylenethiomethoxy (-CH,SCH3), ethyienethio methoxy (-Chi,Ch,SCH3), n-propylene-is¢-thiopropoxy.s (-CH,CH,CH,SCH(CH3),), methylene--thiobutoxy (-CH,SC(CH3D3) and the like.

The term "alkenyl" refers to a monoradical of a branched or unbranched unsaturated hydrocarbon group preferably having from 2 to 40 carbon atoms, more preferably 2 to 10 carbon atoms and even more preferably 2 tos 6 carbon atoms and having at least 1 and preferably from 1-6 sites of vinyl urmsaturation.

Preferred alkenyl groups include ethenyl (-CH=CH,), n-propenyl (-CH,CH=CH,), iso-propenyl (-C(CH3)=CH,), and the like.

V0 99/64042 PCT/US99/12728 ’

The term "substituted alkenyl" refers to an alkenyl group as defined above having from 1 to 5 substituents, and preferably 1 to 3 substituents, selected from the group consisting of alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl. acy 1, acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy, oxyarninoacyl, azido, cyano, halogen, hydroxyl, keto, thioketo, carboxyl, carboxylalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, heterocyclic, heterocyclooxy, hydroxyamino, alkoxyamino, nitro, -SO-alkyl, -SO-substituted alkyl, -SO-aryl, -

SO-heteroaryl, -5SO,-alkyl, -SO,-substituted alkyl, -SO,-aryl and -SO,-heteroaryl. . The term "alkenylene" refers to a diradical of a branched or unbranched unsaturated hydrocarbon group preferably having fronn 2 to 40 carbon atoms, more preferably 2 to 10 carbon atoms and even more preferably 2 to 6 carbon atoms and having at least 1 and preferably from 1-6 sites of vinyl unsaturation.

This term is exemplified by groups such as ethenylene (-CH=CH-), the propenylene isomers (e.g., -CH,CH=CH- and -C(CH 3)=CH-) and the like.

The term "substituted alkenylene" refers to an alkenylene group as defined above having from 1 to 5 substituents, and preferably from 1 to 3 substituents, selected from the group consisting of alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl, keto, thioketo, carboxyl, carboxylalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, heterocyclic, heterocyclooxy, hydroxyamino, alkoxyamino, nitro, -SO-alkyl, -SO-substituted alkyl, -SO-aryl, -

Additionally, such substituted alkenylene groups inclucle those where 2 substituents on the alkeenylene group are fused to form one or more cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl, heterocyclic or heteroaryl groups fusead to the alkenylene group.

The term "alky nyl" refers to a monoradical of an unsaturated hydrocarbon preferably having from 2 to 40 carbon atoms, more preferably 2 to 20 carbon atoms and even more preferably 2 to 6 carbon atoms and having at least 1 and preferably from 1-6 sikes of acetylene (triple bond) unsaturation. Preferred alkynyl groups include ethynyll (-C=CH,), propargyl (-CH,C=CH) and the like.

The term "substituted alkynyl” refers to an alkynyl group as defined above having from 1 to 5 substituents, and preferably 1 to 3 substituents, selected from - the group consisting o f alkoxy, substituted alkoxy, cycloalkyl, substituted : cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, substituted am3no, aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, } halogen, hydroxyl, keto, thioketo, carboxyl, carboxylalkyl, thioaryloxy, 3 15 thioheteroaryloxy, thioheterocyclooxy, thiol, thicalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroamyl, heteroaryloxy, heterocyclic, heterocyclooxy, hydrexyamine, alkexy=amino, nitre, -SQ-alky!, -SO-substituted alkyl, -SO-aryl, -

SO-heteroaryl, -SO,-a lkyl, -SO,-substituted alkyl, -SO,-aryl and -SO,-heteroaryl.

The term "alky=nylenc" refers to a diradical of an unsaturated hydrocarbon preferably having frorm 2 to 40 carbon atoms, more preferably 2 to 10 carbon atoms and even more ppreferably 2 to 6 carbon atoms and having at least 1 and preferably from 1-6 sites of acetylene (triple bond) unsaturation. Preferred alkynylene groups incBude ethynylene (-C=C-), propargylene (-CHC=C-) and the like.

The term "substituted alkynylene” re fers to an alkynylene group as defined above having from 1 to 5 substituents, and ppreferably 1 to 3 substituents, selected from the group consisting of alkoxy, substitLated alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl, amino acyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl, keto, thioketo, carboxyl, carboxylalkyl, thicaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol , thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, het erocyclic, heterocyclooxy, hydroxy- amino, alkoxyamino, nitro, -SO-alkyl, -SO-substituted alkyl, -SO-aryl, -SO- heteroaryl, -SO,-alkyl, -SO,-substituted alkyl, -SO,-aryl and -SO,-heteroaryi.

The term "acyl" refers to the groups HC(O)-, alkyl-C(O)-, substituted alkyl-C(O)-, cycloalkyl-C(O)-, substituted cwcloalkyl-C(O)-, cycloalkenyl-C(O)-, substituted cycloalkenyl-C(O)-, aryl-C(O)-, heteroaryl-C(O)- and heterocyclic-

C(O)- where alkyl, substituted alkyl, cycloal kyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl, heteroaryl and heterocyclic are as defined herein.

The term "acylamino" refers to the gmoup -C(O)NRR where each R is independently hydrogen, alkyl, substituted allkyl, aryl, heteroaryl, heterocyclic or where both R groups are joined to form a heterocyclic group (e.g., morpholino) wherein alkyl, substituted alkyl, aryl, heterozaryl and heterocyclic are as defined herein.

The term "aminoacyl” refers to the group -NRC(O)R where each R is independently hydrogen, alkyl, substituted al kyl, aryl, heteroaryl, or heterocyclic wherein alkyl, substituted alkyl, aryl, heteroaryl and heterocyclic are as defined herein.

The term "aminoacyloxy" refers to the group -_-NRC(O)OR where each R is independently hydrogen, alkyl, substituted alkyl, aryl, heteroaryl, or heterocyclic wherein alkyl, substituted alkyl, aryl, heteroaryl and heterocyclic are as defined herein.

The term "acyloxy" refers to the groups alkyl-C(0Q)O-, substituted alkyl-

C(0)O-, cycloalkyl-C(0O)O-, substituted cycloalkyl-C(C)O-, aryl-C(0)O-, heteroaryl-C(0)O-, and heterocyclic-C(O)O- wherein alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, heteroaryl, and heterocyclic are as defined herein.

The term "aryl" refers to an unsaturated aromatic carbocyclic group of from 6 to 20 carbon atoms having a single ring (e.g., pshenyl) or multiple condensed (fused) rings (e.g., naphthyl or anthryl). Pr eferred aryls include phenyl, naphthyl and the like.

Unless otherwise constrained by the definition feor the aryl substituent, such aryl groups can optionally be substituted with from 1 toe 5 substituents, preferably 1103 substituents, selected Tom the group consisting cif acyioxy, hydroxy, thiol, acyl, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, substituted alkyl, substituted alkoxy, substituted alkenyl, substituted alkyryl, substituted cycloalkyl, substituted cycloalkenyl, amino, substituted amino, amimoacyl, acylamino, alkaryl, aryl, aryloxy, azido, carboxyl, carboxylalkyl, cyano, halo, nitro, heteroaryl, heteroaryloxy, heterocyclic, heterocyclooxy, aminoacyloxy, oxyacylamino, thioalkoxy, substituted thioalkoxy, thioaryloxy, thiohete roaryloxy, -SO-alkyl, -SO- substituted alkyl, -SO-aryl, -SO-heteroaryl, -SO,-alkyl, -SO,-substituted alkyl, - 50,-aryl, -SO,-heteroaryl and trihalomethyl. Preferred aryl substituents include alkyl, alkoxy, halo, cyano, nitro, trihalomethyl, and thicalkoxy.

The term "aryloxy" refers to the group aryl-O- wherein the aryl group is as defined above including optionally substituted aryl groups as also defined above.

The term “arylene” refers to the diradical derived from aryl (including substituted aryl) as defined above and is exemplified by 1,2-phenylene, 1,3- phenylene, 1,4-phenylene, 1,2-naphthylene and the like.

The term “amino” refers to the group -NH,.

The term “substituted amino” refers to the group -NRR where each R is independently selected from the group consisting of hydrogen, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, alkenyl, substituted alkenyl, cycloalkenyl, substituted cycloalkenyl, alkynyl, substituted alkynyl, aryl, heteroaryl and heterocyclic provided that both R's are not hydrogen.

The term "carboxyalkyl" refers to the groups "-C(O)O-alkyL", “-C(O)O- substituted alkyl”, “-C(O)O-cycloalkyl”, “-C(0)O-substituted cycloalkyl”, “-C(0)O-al kenyl”, “~C(O)O-substituted alkenyl”, “-C(0)O-alkyny}” and “-C(0)O-s.abstituted alkynyl” where alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, alkenyl, substituted alkenyl, alkynyl and sub stituted alkynyl are as defireed herein.

The term "cycloalkyl" refers to cyclic alkyl groups of from 3 to 20 carbon atoms having a single cyclic ring or multiple condensed rings. Such cycloalkyl groups include, by way of example, single ring structures such as cyclopropyl, cyclobutyl, cyclopentyl, cyclooctyl, and the like, or multiple ring structures such as adamantanyl, and the like.

The term "substituted cycloalkyl" refers tos cycloalkyl groups having from 1 to 5 substituents, and preferably 1 to 3 substituents, selected from the group consisting of alkoxy, substituted alkoxy, cycloalkwyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloay, oxy aminoacyl, azido, cyano, halogen, hydroxyl, keto, thivketo, carboxyl, carbeoxylalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, heterocy clic, heterocyclooxy, hydroxyamino, alkoxyamino, nitro, -SO-alkyl, -S<O-substituted alkyl, -SO-aryl, -SO-heteroaryl, -SO,-alkyl, -SO,-substituted alky 1, -SO,-ary} and -SO,-heteroaryl.

The term "cycloalkenyl" refers to cyclic alkenyl groups of from 4 to 20 carbon atoms having a single cyclic ring and at lezast one point of internal unsaturation. Examples of suitable cycloalkenyl groups include, for instance, cyclobut-2-enyl, cyclopent-3-enyl, cyclooct-3-eny 1 and the like.

The term "substituted cycloalkenyl” refers to cycloalkenyl groups having from 1 to 5 substituents, and preferably 1 to 3 sub»stituents, selected from the group consisting oF aikoxy, substituted alkoxy, cy cloaiky:, substituted cycioaikyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl, keto, thioketo, carboxyl, carboxylalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, heterocyclic, heterocyclooxy, hydroxyamino, alkoxyamino, nitro, -SO-alkyl, -SO-substituted alkyl, -SO-aryl, -SO-heteroaryl, -SO,-alkyl, -SO,-substituted alkyl, -SO,-aryl and -SO,-heteroaryl.

The term "halo" or "halogen" refers to fluoro, chloro, bromo and iodo.

The term ®*'heteroaryl” refers to an aromatic group of from 1 to 15 carbon atoms and 1 to 4 Theteroatoms selected from oxygen, nitrogen and sulfur within at least one ring (if here is more than one ring).

Unless othaerwise constrained by the definition for the heteroaryl substituent, such ‘heteroaryl groups can be optionally substituted with 1 to 5 substituents, preferably 1 to 3 substituents, selected from the group consisting of acyloxy, hydroxy-, thiol, acyl, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, sub stituted alkyl, substituted alkoxy, substituted alkenyl, substituted alkynyl, substituted cycloalkyl, substituted cycloalkenyl, amino, substituted amino, aminoacyl, acylarmino, alkaryl, aryl, aryloxy, azido, carboxyl, carboxylalkyl, cyano, halo, nitro, heteroaryl, heteroaryloxy, heterocyclic, heterocyclooxy, aminoacyloxy, oxyacylamino, thioalkoxy, substituted thioalkoxy, thioaryloxy, thioheteroaryloxy”, -SO-alkyl, -SO-substituted alkyl, -SO-aryl, -SO-heteroaryl, -

SO,-alkyl, -SO,- substituted alkyl, -SO,-aryl, -SO,-heteroary! and trihalomethyl.

Preferred aryl substituents include alkyl, alkoxy, halo, cyano, nitro, trihalomethyl, and thioalkoxy. Such heteroaryl groups can have a single ring (e.g., pyridyl or furyl) or multiple condensed rings (e.g., indolizinyl or benzothienyl). Preferred heteroaryls include pyridyl, pyrrolyl and furyl.

The term “heteroaryloxy” refers to the group heteroaryl-O-.

The term “heteroarylene” refers to the diradical group derived from heteroaryl (includling substituted heteroaryl), as defined above, and is exemplified by the groups 2,6-pyridylene, 2,4-pyridiylene, 1,2-quinolinylene, 1,8- quinolinylene, 1,<4-benzofuranylene, 2,5-pyridnylene, 2,5-indolenyl and the like.

The term “heterocycle” or "heterocyclic" refers to a monoradical saturated unsaturated group having a single ring or multiple condensed rings, from 1 to 40

Unless otherwise constrained by the definitiosn for the heterocyclic substituent, such heterocyclic groups can be optionally substituted with 1 to 5, and preferably 1 to 3 substituents, selected from the group consisting of alkoxy, substistuted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl, aminosacyloxy, oxyaminoacyl, azido, cyano, halogem, hydroxyl, keto, thioketo, carboxyl, carboxylalkyl, thioaryloxy, thioheteroaryleoxy, thioheterocyclooxy, thiol, thioalloxy, substituted thioalkoxy, aryl, aryloxy, hewmeroaryl, heteroaryloxy, hetero-cyclic, heterocyclooxy, hydroxyamino, alkoxy~amino, nitro, -SO-alkyl, -SO- substituted alkyl, -SO-aryl, -SO-heteroaryl, -S0O,-alkzyl, -SO,-substituted alkyl, -

SO,-amryl and -SO,-heteroaryl. Such heterocyclic groups can have a single ring or multiple condensed rings. Preferred heterocyclics include morpholino, piperidinyl, and the like.

Examples of nitrogen heterocycles and hetero.aryls include, but are not limitec= tc, pyrrole, imidazole, pyrazoie, pyridine, pyrazine, pyrimidine, pyrida=ine, indolizine, isoindole, indole, indazole, puarine, quinolizine, isoquirmoline, quinoline, phthalazine, naphthylpyridine, quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline, phenanthriedine, acridine, phenan throline, isothiazole, phenazine, isoxazole, phe=noxazine, phenothiazine, imidazolidine, imidazoline, piperidine, piperazine, incloline, morpholino, piperidinyl, tetrahydrofuranyl, and the like as well as N-alkoxy-nitrogen containing heterocycles. -A preferred class of heterocyclics include “croswn compounds” which refers tO a specific class of heterocyclic compounds having one or more repeating units of the fosrmula [-(CH,-),Y-] where mis >» 2, and Y at each separate occurrence ca.n be O, N, S or P. Examples of crown compounds include, by way of example oraly, [-(CH,);-NH-]3, [-((CH,),-0)4-((CH,),-NIH),] and the like.

Typically such crown compounds can have from 4 to 10 heteroatoms and 8 to 40 carbon atoms

The te=rm “heterocyclooxy” refers to the group heter «ocyclic-O-.

The te=rm “thioheterocyclooxy” refers to the group h eterocyclic-S-.

The term “heterocyclene” refers to the diradical gromp formed from a heterocycle, as defined herein, and is exemplified by the groups 2,6-morpholino, 2,5-morpholi no and the like.

The term "oxyacylamino” refers to the group -OC(C3)NRR where each R 1s independentls hydrogen, alkyl, substituted alkyl, aryl, hetex-oaryl, or heterocyclic wherein alky 1, substituted alkyl, aryl, heteroaryl and hetero <yclic are as defined herein.

The term "thiol" refers to the group -SH.

The term "thioalkoxy" refers to the group -S-alkyl.

The term "substituted thioalkoxy" refers to the grougp -S-substituted alkyl.

The term "thioaryloxy" refers to the group aryl-S- wherein the aryl group is as defined above including optionally substituted aryl gro-ups also defined above.

The term "thioheteroaryloxy" refers to the group heteroaryl-S- wherein the heteroaryl group is as defined above including optionally substituted aryl groups as also defined above.

As 10 any of the above groups which contain one or more substituents, it is understood, of course, that such groups do nost contain any substitution or substitution patterns which are sterically impr actical and/or synthetically non- feasible. In addition, the compounds of this invention include all stereochemical isomers arising from the substitution of these compounds.

The term "pharmaceutically acceptable salt" refers to salts which retain the biological effectiveness and properties of the mmulti-binding compounds of this invention and which are not biologically or ot_herwise undesirable. In many cases, the multi-binding compounds of this inventioma are capable of forming acid and/or base salts by virtue of the presence of amino and/or carboxyl groups or groups similar thereto.

Pharmaceutically acceptable base addition salts can be prepared from incrganic anc organic bases. Salts derived from inorganic bases, inciude dy way of example only, sodium, potassium, lithium, ammonium, calcium and magnesium salts. Salts derived from organic bases includee, but are not limited to, salts of primary, secondary and tertiary amines, such zas alkyl amines, dialkyl! amines, trialkyl amines, substituted alkyl amines, di(sumbstituted alkyl) amines, tri(substituted alkyl) amines, alkenyl amines, cialkenyl amines, trialkenyl amines, substituted alkenyl amines, di(substituted alkeryl) amines, tri(substituted alkenyl) amines, cycloalkyl amines, di(cycloalkyl) amiraes, tri(cycloalkyl) amines, substituted cycloalkyl amines, disubstituted cyecloalkyl amine, trisubstituted cycloalkyl amines, cycloalkenyl amines, di(.ycloalkenyl) amines, tri(cycloalkenyl) amines, substituted cycloalkenyl amines, disubestituted cycloalkenyl amine,

trisubstituted cycloalkenyl amines, aryl amines, diaryl amines, triaryl amines, heteroaryl amines, dilmeteroaryl amines, triheteroaryl amines, heterocyclic amines, diheterocyclic amines ,. triheterocyclic amines, mixed di- and tri-amines where at least two of the substimuents on the amine are different and are selected from the group consisting of alkyl, substituted alkyl, alkenyl, substituted alkenyl, cycloalkyl, substitute cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl, heteroaryl, heterocyclic, and the like. Also included are amines where the twoo r three substituents, tog-cther with the amino nitrogen, form a heterocyclic or heteroaryl group.

Examples of suitable amines include, by way of example only, isopropylamine, trime=thy! amine, diethyl amine, tri(iso-propyl) amine, tri(n- propyl) amine, ethano-lamine, 2-dimethylaminoethanol, tromethamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine, gluc osamine, N-alkylglucamines, theobromine, purines, piperazine, piperidine , morpholine, N-ethylpiperidine, and the like. It should also be understood that otlmer carboxylic acid derivatives would be useful in the practi ce of this invention, for &xample, carboxylic acid amides, including carboxamides, lower alkyl carboxamades, dialkyl carboxamides, and the like.

Pharmaceutically acceptable acid addition salts may be prepared from inorganic and organic acids. Salts derived from inorganic acids include hydrochloric acid, hycrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like. Salts der-ived from organic acids include acetic acid, propionic acid , glycolic acid, pyruvic acid, oxalic acid, malic acid, malonic acid, succinic acid, maleic acid, fumaric a.cid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, metharmesulfonic acid, ethanesulfonic acid, p-toluene-sulfonic acid , salicylic acid, and the like.

The= term "protecting group" or "blocking group" refers £0 any group which whe=n bound to one or more hydroxyl, thiol, amino or carboxyl groups of the compowunds (including intermediates thereof) prevents reactions from occurring at these greoups and which protecting group can be removed by c-onventional chemical osr enzymatic steps to reestablish the hydroxyl, thiol, armino or carboxyl group.’ T he particular removable blocking group employed is mot critical and preferred removable hydroxyl blocking groups include conventional substituents such as allZyl, benzyl, acetyl, chloroacetyl, thiobenzyl, benzylidire, phenacyl, t- butyl-diphenylsilyl and any other group that can be introduced ckaemically onto a hydroxyl faunctionality and later selectively removed either by ch_emical or enzymatic methods in mild conditions compatible with the nature of the product.

Pre=ferred removable amino blocking groups include conv-entional substituent s such as t-butyoxycarbonyl (t-BOC), benzyloxycarbomyl (CBZ), and the like whaich can be removed by conventional conditions compatible with the nature of tke product.

Preferred carboxyl protecting groups include esters such zs methyl. ethyl, £ropyl, e-bauty! ete. which can be remeved by mild Lycrolysis comditions compatible with the nature of the product.

The: term "optional" or "optionally" means that the subsequently described event, circLamstance or substituent may or may not occur, and thamt the description includes insstances where said event or circumstance occurs and irstances where it does not.

As 1ased herein, the terms “inert organic solvent” or “inert solvent” mean a solvent iner-t under the conditions of the reaction being described in conjunction therewith [i ncluding, for example, benzene, toluene, acetonitrile, tetrahydrofuran

(“"THF”), dimethylformamide (“DMF”), chloroform (CHCl,), methylene chloride (ox dichloromethane or “CH,Cl,"), diethyl ether, ethyl &acetate, acetone, me=thylethy] ketone, methanol, ethanol, propanol, isoprogpanol, tert-butanol, dioxane, pyridine, and the like]. Unless specified to the contrary, the solvents us-ed in the reactions of the present invention are inert so-lvents. “Benign prostatic hyperplasia” (“BPH”) refers to» nonmalignant enlargement of the prostate that is directly attributable toe cellular hyperplasia of booth glandular and stromal elements of the gland. It is diagnosed histologically, de=veloping in the periuretheral transition zone of the pro state.” BPH progresses fresquently to benign prostatic enlargement and causes be-nign prostatic obstruction armd lower urinary tract symptoms, often referred to as ssymptomatic BPH.” BPH is currently the most prevalent urological disease in men 3 Current treatment for

BE’H include surgery, ultrasound and androgen-suppress ing therapy.’

The “alpha-1A adrenergic receptor,” also known as alpha-1A adLrenoreceptor, is a G-protein coupled transmembrane receptor that is responsible for prostate smooth muscle contraction.’ The alpha-1A adrenergic receptor is pr esent in the greatest concentration in the human prostate’ and smooth vascular muscle tissue. Antagonists of this receptor relax the smooth muscles of the pr-ostate, for example, thereby decreasing bladder outlet resistance and facilitating urinary flow without affecting detrusor smooth muscle contractility.’

It should be recognized that the alpha-1A adrener-gic receptors that participate in biological multivalent binding interactions arc constrained to varying de grees by their intra- and intermolecular associations (e .g. cellular receptors may be covalently joined in a single structure, noncovalently associated in a multimeric structure, embedded in a membrane or polymeric matrix and so on) and therefore

W060 PCT/US99/12728 have less translational and rotational freedom th an if the same cellular receptors were present as monomers in solution.

The term "library" refers to at least 3, pmreferably from 102 to 109 and more preferably from 102 to 104 multimeric cormpounds. Preferably, these compounds are prepared as a multiplicity of comnpounds in a single solution or reaction mixture which permits facile synthesis thereof. In one embodiment, the library of multimeric compounds can be directly assayed for multibinding properties. In another embodiment, each memb er of the library of multimeric compounds is first isolated and, optionally, characterized. This member is then assayed for multibinding properties.

The term “collection” refers to a set of multimeric compounds which are prepared either sequentially or concurrently (e.g-., combinatorially). The collection comprises at least 2 members; prefera bly from 2 to 109 members and still more preferably from 10 to 104 members.

The term “ligand binding site” as used herein denotes the site on the alpha- 1A adrenergic receptor that recognizes a .iganc ciomain and provides a binding partner for that ligand. The ligand binding site rmay be defined by monomeric or multimeric structures. This interaction may be capable of producing a unique biological effect, for example agonism, antagonism, modulatory effect and the like or may maintain an ongoing biological event. “Ligand” or “alpha-1A adrenergic ligand ” as used herein denotes a compound that is a binding partner for the alpha- 1A adrenergic receptor and is bound thereto by complementarity. The specific region or regions of the ligand that is (are) recognized by the alpha-1A adrenergic receptor is designated as the “ligand binding domain”. A ligand may be either capable of binding to a receptor by itself, or may reequire the presence of one or more non-ligand cormponents for binding (e.g., cat? Mgt? or a water molecule).

It is further understood that the term “ligand” or “alpha-1A adrenergic ligand” is not intemnded to be limited to compounds known to be useful as alpha-1A adrenergic recepto-t binding compounds (e.g., known drugs). It should also be understood that posrtions of the ligand structure that are not essential for specific molecular recognition and binding activity may be varied substantial ly, replaced with unrelated structures and, in some cases, omitted entirely withouat affecting the binding interactiora. The primary requirement for the ligand is that &t has a ligand domain as defined above. Those skilled in the art will understand thaat the term ligand can equally apply to a molecule that is not normally associate d with alpha- 1A adrenergic cell ular receptor binding properties. In addition, it should be noted that ligands that exchibit marginal activity or lack useful activity as monomers can be highly active as= multivalent compounds because of the benefits conferred by multi-valency. Th e only requirement for a ligand is that it has a ligand binding domain as defined above.

Accordingly, examples of ligands useful for this invention in«<lude terazosin, prazosimm, doxasosin, alfuzosin, tamsulosin, RS 100975 (Roche

Biosciences), A-131701 (Abbott), L794-191 (Merck), L757464 (Mexck), REC 15- 2739 (SB216469, Recordati/SKB), KMD-3213 (Kissei Pharmaceuticals), and derivatives thereof . Each of these ligands are known to be useful in the treatment of BPH and/or hypeertension. )

A “multime=ric compound” refers to a compound that may be capable of multivalency as defined below, and which has 2 to 10 ligands covalently bound to one or more linkerss which may be the same or different. The compound may or may not possess multibinding properties. At least one of the ligands comprises a ligand binding domain capable of binding to an alpha-1A adrenergic rece=ptor. The multi-binding compound provides a biological and/or therapeutic effect goreater than the aggregate of unlinked ligands equivalent thereto which may be ®he same or different which unlinked ligands comprise a ligand domain capable of binding to one or more alpha-1A adrenergic receptors. That is to say that the biological and/or therapeutic effect of the alpha-1A adrenergic binding ligands attac-hed to the multi-binding compound is greater than that achieved by the same amoun_t of unlinke=d alpha-1A adrenergic ligands made available for binding to the li gand binding sites.

The phrase “increased biological or therapeutic effect” includes, for example increased affinity for a target, increased specificity for a target, mncreased selectiwity for a target, increased potency, increased efficacy, decreased teoXicity, improved duration of action, decreased side effects, increased therapeutic index, improv-ed bioavailability, improved pharmacokinetics, improved activity Spectrum, and thes like. The multi-binding compounds of this invention will exhibit at least one and preferably more than one of the above mentioned effects. “Uri-vaiency” as usec herein refers to a single binding interaction between one ligand as defined herein with one ligand binding site as defined hereirm. It should Ibe noted that a molecule having multiple copies of a ligand (or liga_nds) exhibits uni-valency when only one ligand is interacting with a ligand binding site.

Exampl es of a univalent interaction are depicted below.

“Multi-valency” as used herein refers to the concurresnt binding of from 2 to 10 linked ligands (which may be the same or different) and two or more corresponding ligand binding sites on the receptors which re ceptors may be the same or different.

For example, two ligands connected by a linker that bind concurrently to two ligand binding sites would be considered as bi-valency; three ligands thus connected would be an example of tri-valency. An example of tri-valency illustrating a multi-binding agent bearing three ligands vers s 2 monovalent binding interaction is shown below: = — & univalent interaction trivalent interaction

It should be understood that all compounds that contain multiple copies of a ligand attached to a linker do not necessarily exhibit the phenomena of multi- valency, i.e., that the biological and/or therapeutic effect of the multi-binding as agent is greater than the sum of the aggregate of unlinked ligzands made available to the ligand binding site. For multivalency to occur, the ligzands that are connected by a linker have to be presented to their receptors by the linker in a specific manner in order to bring about the desired ligand-or ienting result, and thus produce a multi-binding agent.

“Potency” as used herein refers to the minimmzam concentration at which a ligand is able to achieve a desirable biological or therzapeutic effect. The potency of a ligand is typically proportional to its affinity for its ligand binding site. In some cases the potency may be non-linearly correlatecd with its affinity. In comparing the potency of two drugs, e.g., a multi-bimding agent and the aggregate of its unlinked ligand, the dose-response curve of each is determined under identical test conditions (e.g. an in vitro or in vivo ass ay, in an appropriate animal model such as a human patient). The finding that the multi-binding agent produces an equivalent biological or therapeutic effect at a lowesr concentration than the aggregate unlinked ligand (e.g. on a per weight, per mole or per ligand basis) is indicative of enhanced potency. “Selectivity” or “specificity” is a measure of tthe binding preferences of a ligand for different ligand binding sites. The selectivi ty of a ligand with respect to its target ligand binding site relative to another ligand binding site is given by the ratio of the respective values of Kj, (i.e., the dissociati on constants for each ligand- receptor complex) or in cases where a biological effect is observed below the Ky. the ratio of the respective ECs s (i.c., the concentrations that produce 50% of the maximum response for tie ligand interacting with the Iwo distinc: iigand binding sites).

The terms "agonism" and "antagonism" are well known in the art. The term "modulatory effect” refers to the ability of the lig-and to change the activity of an agonist or antagonist through binding to a ligand birding site.

The term “partial agonist” refers to a receptor zagonist which cannot fully elicit a maximal response when it binds to the receptor, no matter how high the concentration of the partial agonist. A parti. agonist i.s able to combine with the receptor, but the full effect of the binding is not elicited. This term is well known in the art andl a discussion of it may be found in Textbook of Receptor

Pharmacolog:y, ch 1.4, J. Foreman and T. Johansen eds., CRC Press, 1996.

The term “treatment” refers to the treatment of benign prostatic hyperplasia (BPH) and/ox hypertension, particularly in a human, and includes: (1) alleviating the symptoms of BPH and/or hypertension; (ii) inhibiting BPH and/or hypertension by prophylactic €reatment; or (iii) relieving or reducing BPH and/or hypertension, e.g. , relieving or reducing the duration, intensity and/or severity of the condition.

The term "therapeutically effective amount” refers to that armount of multi- binding com pound which is sufficient to effect treatment, as defined above, when administered to a mammal in need of such treatment. The therapeu tically effective amount will vary depending upon the subject and disease condition being treated, the weight amd age of the subject, the severity of the disease condition, the manner of administration and the like, which can readily be determined by ©ne of ordinary skill in the a rt.

The term "linker," identified where appropriate by the symbol “X”, refers to a group ox groups that covalently link(s) from 2 to 10 ligands (as identified above) in a rmanner that provides for a compound capable of multi-walency when in the presence of at least one cellular receptor having 2 or more ligarad binding sites.

The linker iss a ligand-orienting entity which may be chiral or achiral that permits attachment oof multiple copies of a ligand (which may be the same or different) thereto. In seme cases the linker may be biologically active. The texm linker does not, howeve 1, extend to cover solid inert supports such as beads, ghass particles, fibers and thme like. But it is to be understood that the multi-bindings compounds of this inventio m can be attached to a solid support if desired, for example, for use in separation amd purification processes and for similar applications.

The ligands and linkers which cormprise the multibinding agents of the invention and the multibinding compounds themselves may have various stereoisomeric forms, including enantiormers and diastereomers. It is to be understood that the invention contemplates all possible stereoisomeric forms of multibinding compounds, and mixtures timereof.

The extent to which multivalent binding is realized depends upon the efficiency with which the linker or linkers that joins the ligands presents them to their ligand binding sites on one or more receptors. Beyond presenting ligands for multivalent interactions with ligand binding sites, the linker spatially constrains these interactions to occur within dimensi ons defined by the linker. Thus the structural features of the linker (valency, geometry, orientation, size, flexibility, chemical composition) are features of mulltivalent compounds that play an important role in determining their activit ies.

Methodology

The linker, when covalently attacheed to the ligands, provides a biocompatible, substantially non-immunogenic mulii-binding compound of this icveaticr. The biological activity of the mnulti-binding compound is highly sensitive to the valency, geometry, compoesition, size, flexibility or rigidity, etc. of the linker as well as the presence or absen ce of anionic or cationic charge, the relative hydrophobicity/hydrophilicity of the linker, and the like on the linker. In general, the linker may be chosen from an y organic molecule construct that orients two or more ligands to the receptors to pexrmit multi-valency. In this regard, the linker can be considered as a "framework'® on which the ligands are arranged in order to bring about the desired ligand-orienting result, and thus produce a multi- binding compound.

Ancillary groups which enhance the water solubility/hydrophilicity of the linker and, accordingly, the resulting multi-binding compounds are useful in practicing this invention. Thus, it is within the scope of the present invention to use ancillary groups such as, for example, poly(ethylene glycols), alcohols, polyols, (e.g., glycerin, glycerol propoxylate, saccharides, including mono-, oligo- and polysaccharides, etc.) carboxylates, polycarboxylates, (e.g., polyglutamic acid, polyacrwlic acid, etc.), amines, polyamines, (e.g., polylycine, poly(ethyleneimine), and the like) to enhance the water solubility and/or hydrophilicity of the multi—binding compounds of this invention. In preferred embodiments, the ancillary group used to improve water solubility/hydrophilicity will be a polyether. In par ticularly preferred embodiments, the ancillary group will be a poly(ethylene glycol).

The incorporation of lipophilic ancillary groups within the structure of the linker to enhance the lipophilicity and/or hydrophobicity of the multi-binding compounds described herein is within the scope of this invention. Lipophilic groups useful with the link ers of this invention include, by way of example only, aryl and heteroaryl groups which, as above, may be either unsubstituted or substituted with other grou ps, but are at least substituted with a group which allows their covalent attactament to the linker. Other lipophilic groups useful with the linkers of this invention include fatty acid derivatives which do not form bilayers in aqueous mediurm until higher concentrations are reached.

Also within the scope of this invention is the use of ancillary groups which result in the multi-binding compound being incorporated into a vesicle such as a liposome or a micelle. The term "lipid" refers to any fatty acid derivative that is capable of forming a bilayer such that a hydrophobic portion of the lipid material orients toward the bilayer swhile a hydrophilic portion orients toward the aqueous phase. Hydrophilic charac teristics derive from the presence of phosphato,

carboxylic, sulfato, amino, sulfhydryl, nitro and other like groups well known in the art. Hydrophobicity could be conferred by the inclusion of groups that include, but are not limited to, long chain saturated and unsaturated aliphatic hydrocarbon groups of up to 20 carbon atoms and such groups substituted by one or more aryl, heteroaryl, cycloalkyl, and/or heterocyclic group(s). Preferred lipids are phosphoglycerides and sphingolipids, representative examples of which include phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, phosphatidic acid, palmitoyleoyl phosphatidylcholine, lysophosphatidyicholine, lysophosphatidy!-ethanolamine, dipalmitoylphosphatidylcholime, dioleoylphosphatidylcholine, distearoyl- phosphatidylcholine or dilinoJeoylphosphatidylcholine could be used. Other compounds lacking phosphorus, such as sphingolipid and glycosphingolipid families are also within the group designated as lipid. Additionally, the amphipathic lipids described above may be mixed with other lipids including triglycerides and sterols.

The flexibility of the 1 inker can be manipulated by the inclusion of ancillary groups which are bulky and/or rigid. The presence of bulky or rigid groups can hinder free rotation about borads in the linker cr bonds between the linker anc the ancillary group(s) or bonds between the linker and the functional groups. Rigid groups can include, for example, those groups whose conformational lability is restrained by the presence of xings and/or multiple bonds, for example, aryl, heteroaryl, cycloalkyl and heterocyclic groups. Other groups which can impart rigidity include polypeptide groups such as oligo- or polyproline chains.

Rigidity can also be imaparted electrostatically. Thus, if the ancillary groups are either positively or negatively charged, the similarly charged ancillary groups will force the presenter linker into a ~onfiguration affording the maximum distance between each of the like charges. The energetic cost of bringing the like-

charged groups closer 10 each other will tend to hold the linker in a configuration that maintains the separation between the like-charged ancillary groups. Further ancillary groups bearing opposite char ges will tend to be attracted to their oppositely charged counterparts and potentially may enter into both inter- and intramolecular ionic bonds. This non-covalent mechanism will tend to hold the linker into a conformation which allows bonding between the oppositely charged groups. The addition of ancillary groups which are charged, or alternatively, bear a latent charge when deprotected, following the addition to the linker, include deprotectation of a carboxyl, hydroxyl, thiol or amino protecting group, by a - 10 change in pH, oxidation, reduction or other mechanisms known to those skilled in the art, is within the scope of this invention.

Bulky groups can include, for example, large atoms, ions (e.g., iodine, sulfur, metal ions, etc.) or groups containing large atoms, polycyclic groups, including aromatic groups, non-aromatic groups and structures incorporating one or more carbon-carbon multiple bonds (i.e., alkenes and alkynes). Bulky groups can also include oligomers and polymers which are branched- or straight-chain species. Species that are branched are expected to increase the rigidity of the structure more per unit molecular weight gain than are straight-chain species.

In preferred embodiments, rigidity is imparted by the presence of cyclic groups (e.g., aryl, heteroaryl, cycloalkyl, heterocyclic, etc.). In still further preferred embodiments, the ring is an ary! group such as, for example, phenyl or naphthyl. In other preferred embodiments, the linker comprises one or more six- membered rings or crown groups which, while not rigid, retain the conformation of the linker through conformational eratropy.

In view of the above, it is apparent that the appropriate selection of a linker group providing suitable orientation, entropy and physico-chemical properties is

As explained above, the multi-binding compounds described herein comprise 2-10 ligands for alpha- 1A adrenergic attached to a linker that links the ligands in such a manner that they are presented to the alpha-1A adrenergic receptor for multivalent interactions. The linker spatially constrains these interactions to occur within dimemsions defined by the linker, thus greatly increasing biological activity of the multi-binding compound as compared to the same number of ligands used in rmono-binding form.

The multi-binding comporands of this invention are preferably represented by the empirical formula L)pX)g where L, X, p and g arc as defined above. This is intended to include the several ways in which the ligands can be linked together in order to achieve the objective Of multi-valency, and a more detailed explanation is described below.

As noted previously, the linker may be considered as a framework to which :lgancs are attached. Thus, it shoulc de recognized that the ligands can be attached at any suitable position o n this framework, for example, at the termini of a linear chain or at any intermediate position.

The simplest and most preferred multi-binding compound is a bivalent compound which can be represent ed as L-X-L, where L is a ligand and is the same or different and X is the linker. As trivalent compound could also be represented in a linear fashion, i.e., as a sequenc e of repeated units L-X-L-X-L, in which L is a ligand and is the same or different at each occurrence, as can X. However, a trimer can also be a multi-binding compound comprising three ligands attached to a central core, and thus represented as (L)3X, where the linker X could include,

for example, an aryl or cycloalkyl group. Tetravalent compounds can be represented as, for example, in al inear array:

IL -X-L-X-L-X-L or in a tetrahedral array: " ya 7 *

L L where X and L are as defined herein.

The same considerations apply to higher multibinding compounds of this invention containing 5-10 ligands. However, for multibinding agents attached to a central linker such as aryl or cycloalkyl, there is a self-evident constraint that there must be sufficient attachment sites on the linker to accommodate the number of ligands present; for example, a bemzene ring could not directly accommodate more than 6 ligands, whereas a multi-rinag linker (e.g., biphenyl) could accommodate a larger number of ligands.

Certain of the above described compounds may alternatively be represented as cyclic chains of the form:

Sa

X X

_/ and variants thereof.

) WC 99/64042 PCT/US99/12728

All of the above variations are intende=d to be within the scope of the invention defined by the formula L)pX)g-

In view of the above description of the linker, it is understood that the term “linker” when used in combination with the erm “multibinding compound” includes both a covalently contiguous single Blinker (e. g., L-X-L) and multiple covalently non-contiguous linkers (L-X-L-X- L) within the muitibinding compound. 7 . r Multibindine C i

The multibinding compounds of this imvention can be prepared from readily available starting materials using the followirmg general methods and procedures. It will be appreciated that where typical or prefserred process conditions (i.e., reaction temperatures, times, mole ratios of reactants, solvents, pressures, etc.) are given, other process conditions can also be ussed unless otherwise stated. Optimum reaction conditions may vary with the particu lar reactants or solvent used, but such «conditions can be determined by one skilled imn the art by routine optimization porocedures.

Additionally, as wiii be apparent to theose skiiied in the art, conventionai porotecting groups may be necessary to prevent certain functional groups from uandergoing undesired reactions. The choice Of a suitable protecting group for a particular functional group as well as suitable conditions for protection and eprotection are well known in the art. For example, numerous protecting groups, amnd their introduction and removal, are described in T. W. Greene and G. M.

Nuts, Protecting Groups in Organic Synthesis, Second Edition, Wiley, New

Work, 1991, and references cited therein.

Any compound which acts as a ligand toward the alpha-1A adrenergic receptor can be used as a ligand in this inventi on. It is desirable that the ligand or ligands be antagonists or partial agonists in order to moderate the activity of the alpha-1A adrenergic receptor or lessen or alleviate the condition of BPH and/or hypertension.

Ligands useful in the compound of this invention include ter-azosin, prazosin, doxasosin, a 1fuzosin, tamsulosin, RS 100975 (Roche Biosciences), A- 131701 (Abbott), L79=4-191 (Merck), L757464 (Merck), REC 15-2 739 (SB216469, Recordati/SKB), KMD-3213 (Kissei Pharmaceuticals), and derivatives thereof. These ligand=s may be combined to form homo- or heteroreers as described herein. In particular, a multimer may be formed by joining any ligand selected from the groump terazosin, prazosin, doxasosin, alfuzosin, t amsulosin, RS 100975 (Roche Bioscieences), A-131701 (Abbott), L794-191 (Merck), L757464 (Merck), REC 15-273 9 (SB216469, Recordati/SKB), KMD-3213 (Kissei

Pharmaceuticals), and derivatives thereof with any one or more ligands selected from the group terazossin, prazosin, doxasosin, alfuzosin, tamsulosi n, RS 100975 (Roche Biosciences), AA-131701 (Abbott), L794-191 (Merck), L757" 464 (Merck),

REC 15-2739 (SB216469, Recordati/SKB), KMD-3213 (Kissei Phaarmaceuticals), and derivatives thereof through a linker. The ligands of the inventi_on will be referred to herein as I_-1 through L-11, wherein each of L-1 to L-1 1 corresponds to a ligand selected from the group terazosin, prazosin, doxasosin, -alfuzosin, tamsulosin, RS 100975 (Roche Biosciences), A-131701 (Abbott), [794-191 (Merck), L757464 (Merck), REC 15-2739 (SB216469, Recordati/'S KB), KMD- 3213 (Kissei Pharmaceuticals), and derivatives thereof. Further, each designation of L-1 through L-11 is meant to include derivatives and analogs of the respective ligand.

Combinatorial Librarie=s

Combinatorial approaches for identifying multimeric compounds which possess multibinding properties will now be discussed.

Specifically, factoms such as the proper juxtaposition of the individual ligands of 2 multibinding compound with respect to the relevant array of birading sites on a target or targets is important in optimizing the interaction of the multibinding compound with its target(s) and to maximize the biological advantage through multivalency. Ome approach is to identify a library of candidate multibinding compounds -with properties spanning the multibinding parameters that are relevant for a particul ar target. These parameters include: (1) the identi ty of ligand(s), (2) the orientation of ligands, (3) the valency of the construct, (4) linker length, (5) linker geometry, (6) linker physical properties, and (7) linker chemical functional groups.

Libraries of multimmeric compounds potentially possessing multibindi ng properties (i.e., candidate multibinding compounds) and comprising a multiplicity of such variables are prepared and these libraries are then evaluated via conventional assays corre sponding to the ligand selected and the multibindirag parameters desired. Considerations relevant to each of these variables are s <t forth below.

A single ligand or set of ligands is (are) selected for incorporation in .o the libraries of candidate mu Jtibinding compounds which library is directed against a particular biological targe t or targets. The only requirement for the ligands chosen is that they are capable of interacting with the selected target(s). Thuis, ligands may be known drugs, modified forms of known drugs, substructures of known drugs or substrates of modified forms of known drugs (which are competent to interact with: the target), or other compounds. Ligands are preferably chosen based o n known favorable properties that may be projected to be carried over to or amplifiexd in multibinding forms. Favorable properties include demonstrated safety and efficacy in human patients, ability to increase insulin

~50- sensitivity, ability to lower serum trigly ceride, cholesterol and/or fatty acid levels, etc. However, it is crucial to note that Migands which display an unfavorable property from among the previous list may obtain a more favorable property through the process of multibinding compound formation; i.e., ligands should not necessarily be excluded on such a basis. For example, a ligand that is not sufficiently potent at a particular target So as to be efficacious in a human patient may become highly potent and efficaciomts when presented in multibinding form.

A ligand that is potent and efficacious bwat not of utility because of a non- mechanism-related toxic side effect may have increased therapeutic index (increased potency relative to toxicity) as a multibinding compound. Compounds that exhibit short in vivo half-lives may have extended half-lives as multibinding compounds. Physical properties of ligarads that limit their usefulness (e.g. poor bioavailability due to low solubility, hydrophobicity, hydrophilicity) may be rationally modulated in multibinding forams, providing compounds with physical properties consistent with the desired uti lity.

Several points are chosen on each ligand at which to attach the ligand to the linker. The selected points on the ligand/linker for attachment are functionalized 10 contain complementary reactive functional groups. This permits probing the effects of presenting the ligands to their receptor(s) in multiple relative orientations, an important multibinding design parameter. The only requirement for choosing attachment points is that att aching to at least one of these points does not abrogate activity of the ligand. Sucka points for attachment can be identified by structural information when available. Alternatively, evaluation of ligand/target binding by nuclear magnetic resonance will permit the identification of sites non- essential for ligand/target binding. See, for example, Fesik, et al., U.S. Patent

No. 5,891,643. When such structural imformation is not available, utilization of structure-activity relationships (SAR) for ligands will suggest positions where

© WO 99/64042 PCT/US99/12728 substamtial structural variations are and are not allowed. In the abs -ence of both structimral and SAR information, a library is merely selected with multiple points of attachment to allow presentation of the ligand in multiple distinct orientations.

Subsequent evaluation of this library will indicate what positions are suitable for attachrment.

It is important to emphasize that positions of attachment thas do abrogate the act ivity of the monomeric ligand may also be advantageously included in candid ate multibinding compounds in the library provided that suchm compounds bear at least one ligand attached in a manner which does not abrogamte intrinsic activity. This selection derives from, for example, heterobivalent i nteractions within the context of a single target molecule. For example, considler a receptor antagomnist ligand bound to its target receptor, and then consider mowdifying this ligand by attaching to it a second copy of the same ligand with a limker which allows the second ligand to interact with the same receptor moleculee at sites proximal to the antagonist binding site, which include elements of t he receptor that are not part of the formal antagonist binding site and/or elements off the matrix surroumding the receptor such as the membrane. Here, the most fa-vorablc orientation for interaction of the second ligand molecule with the r eceptor/matrix may be achieved by attaching it to the linker at a position which abxrogates activity of the igand at the formal antagonist binding site. Another way to consider this is that the SAR of individual ligands within the context of a multibindling structure is often different from the SAR of those same ligands in monomeric form.

The foregoing discussion focused on bivalent interactions of dimeric compo unds bearing two copies of the same ligand joined to a single linker through different attachment points, one of which may abrogate the bindings /activity of the monomeric ligand. It should also be understood that bivalent advamtage may also be atta ined with heterodimeric constructs bearing two different ligands that bind to

VO 99/64042 PCT. /US99/12728 common or different targets. For example, terazosin ancl alfuzosin may be joined to a linker through attachment points which do not abrogzate the binding affinity of the monomeric ligands for their respective receptor sites . The dimeric compound may achieve enhanced affinity for both receptors due to Favorable interactions between the terazosin ligand and elements proximal to th e formal alfuzosin ligand binding site and between the alfuzosin ligand and elements proximal to the formal terazosin ligand binding site. Thus, the dimeric compoumd may be a more potent and selective antagonist of the alpha-1A andregenic receptor and a superior therapy for benign prostatic hyperplasia.

Once the ligand attachment points have been chosen, one identifies the types of chemical linkages that are possible at those points. The most preferred types of chemical linkages are those that are compatible with the overall structure of the ligand (or protected forms of the ligand), readily amd generally formed, stable and intrinsically innocuous under typical chemical and physiological conditions, and compatible with a large number of available linkers. Amide bonds, ethers, amines, carbamaies, ureas, and suifonamicles are but a few examples of preferred linkages. functional groups

In the library of linkers employed to generate the Library of candidate multibinding compounds, the selection of linkers employe=d in this library of linkers takes into consideration the following factors.

Yalency. In most instances the library of linkers is initiated with divalent linkers. The choice of ligands and proper juxtaposition of two ligands relative to their binding sites permits such molecules to exhibit target binding affinities and specificities more than sufficient to confer biological advantage. Furthermore, divale=nt linkers or constructs are also typically of modest size such that they retain the de=sirable biodistribution properties of small molecules .

Linker length. Linkers are chosen in a range of lemgths to allow the spanning of a range of inter-ligand distances that encompa ss the distance preferable for a given divalent interaction. In some instances the preferred distan<e can be estimated rather precisely from high-resolution structural information of targets, typically enzymes and soluble receptor targets. In other instan«es where high-resolution structural information is n.ot available, one can make use of simple models to estimate the maximum distance between binding sites either on adjacent receptors or at different locations on the same receptor. In situati-ons where two binding sites are present on the same target (or target subunit for multisubunit targets), preferred linker distances are 2-20 A, with more prefered linker distances of 3-12 A. In situations where t wo binding sites reside on separate (e.g., protein) target sites, preferred linker dis tances are 20-100 A, with maore preferred distances of 30-70 A.

Linker geometry and rigidity. The combination of” ligand attachment site, linker length, linker geometry, and linker rigidity determime the possible ways in which the ligands of candidate multibinding compounds may be displayed in three dimen sions and thereby presented to their binding sites. Linker geometry and rigidity are nominally determined by chemical compositiom and bonding pattern, which may be controlled and are systematically varied as another spanning function in a multibinding array. For example, linker geometry is varied by attaching two ligands to the ortho, meta, and para positions of a benzene ring, or in cis- or trans-arrangements at the 1,1- vs. 1,2- vs. 1,3- ws. 1,4- positions around a cyclohexane core or in cis- or trans-arrangements at a point of ethylene unsatwmration. Linker rigidity is varied by controlling the mumber and relative energies of different conformational states possible for the linker. For example, a divalent compound bearing two ligands joined by 1,8-octy/! linker has many more degrees of freedom, and is therefore less rigid than a com pound in which the two ligands are attached to the 4,4' positions of a biphenyl linker.

Linker physical properties. The physical properties of linkers are nominally determined by the chemical constitution'and bo nding patterns of the linker, and linker physical properties impact the overall physical properties of the candiciate multibinding compounds in which they are incluaded. A range of linker compositions is typically selected to provide a range of ph ysical properties (hydrophobicity, hydrophilicity, amphiphilicity, polarizati on, acidity, and basicity) in the candidate multibinding compounds. The particular choice of linker physical propexties is made within the context of the physical properties of the ligands they join and, preferably, the goal is to generate molecules witkh favorable properties.

For example, linkers can be selected to avoid those that ar e too hydrophilic or too hydrophobic to be readily absorbed and/or distributed in vivo.

Linker chemical functional groups. Linker chemic al functional groups are selected to be compatible with the chemistry chosen to coranect linkers to the ligands and to impart the range of physical properties sufficient to span initial exami nation of this parameter.

Combinatorial synthesis

Having chosen a set of n ligands (n being determin ed by the sum of the number of different attachment points for each ligand chosen) and m linkers by the process outlined above, a library of (n!)m candidate divalesnt multibinding compounds is prepared which spans the relevant multibindling design parameters for a particular target. For example, an array generated from two ligands, one which has two attachment points (Al, A2) and one which has three attachment

) © WO 99/64042 PCT/US99/12728 -55—~ points (B1, B2, B3) joined in all possible ceombinations provide for at least 15 possible combinations of multibinding compounds:

Al-Al Al-A2 Al-Bl A1-B2 _AI-B3 A2-A2 A2-Bl A2B2

A2-B3 B1-B1 BI-B2 BI1-B3 B2-B2 B2-B3 B3-B3

When each of these combinations is joined by 10 different linkers, a library of 150 candidate multibinding compounds results.

Given the combinatorial nature of th e library, common chemistries are preferably used to join the reactive function-alities on the ligands with complementary reactive functionalities on the linkers. The library therefore lends itself to efficient parallel synthetic methods. The combinatorial library can employ solid phase chemistries well known in the art wherein the ligand and/or linker is attached to a solid support. Alternatively arad preferably, the combinatorial library is prepared in the solution phase. After synthesis, candidate multibinding compounds are optionally purified before assaying for activity by, for example, chromatographic methods (e.g., HPLC). methods

Various methods are used to characterize the properties and activities of the candidate multibinding compounds in the library to determine which compounds possess multibinding properties. Physical constants such as solubility under various solvent conditions and logD/clogD v-alues are determined. A combination of NMR spectroscopy and computational me thods is used to determine low-energy conformations of the candidate multibinding compounds in fluid media. The ability of the members of the library to bind to the desired target and other targets is determined by various standard methods, wvhich include radioligand displacement assays for receptor and ion channel targets, and kinetic inhibition analysis for many crmzyme targets. In vitro efficacy, such as for receptor agonists and antagonists, ion channel blockers, and antimicrobial activity are also determined. Pharmamcological data, including oral absorption, everted gut penetration, other phmarmacokinetic parameters and efficacy data are determined in appropriate models. In this way, key structure-activity relationships are obtained for multibinding des@gn parameters which are then used to direct future work.

The members of the library which exhibit multibinding properties, as defined herein, can bse readily determined by conventional methods. First, those members which exhibit multibinding properties are identified by conventional methods as described above, including conventional assays (both in vitro and in vivo).

Second, ascertaining the structure of those compounds which exhibit multibinding properties can be accomplished via art recognized procedures. For example, each membwer of the library can be encrypted or tagged with appropriate information allowing- determination of the structure of relevant members at a later time. See, for exampple, Dower, et al., International Patent Application

Publication No. WO 93/06121; Brenner, et al., Proc. Natl. Acad. Sci., USA, 89:5181 (1992); Gallop, et al., U.S. Patent No. 5,846,839; each of which is incorporated herein by reference in its entirety. Alternatively, the structure of relevant multivalent compounds can also be determined from soluble and untagged libraries of candidates multivalent compounds by methods known in the art, such as those described by Hl indsgaul, et al., Canadian Patent Application No. 2,240,325 which was published. on July 11, 1998. Such methods couple frontal affinity : 25 chromatography witha mass spectroscopy to determine both the structure and relative binding affimities of candidate multibinding compounds to receptors.

) © WO 99/64042 PCT/US99/12728

The process set forth above for dimeric candidate multibinding compounds can, of course, be extended to trimeric candidate compounds and higher analogs thereof.

Follow- hesis and analysis of ad ditional 5

Based on the information obtained through analysis of the initial library, an optional component of the process is to ascertain one or more promising multibinding "lead" compounds as defined by particular relative ligand orientations, linker lengths, linker geornmetries, etc. Additional libraries can then be generated around these leads to provide for further information regarding structure to activity relationships. These arrays typically bear more focused variations in linker structure to further optimize target affinity and/or activity at the target (antagonism, partial agonism , etc.), and/or alter physical properties. By iterative redesign/analysis using the nowel principles of multibinding design along with classical medicinal chemistry, biochemistry, and pharmacology approaches, one is able to prepare and identify optirnal multibinding compounds that exhibit biological advantage towards their targets and as therapeutic agents.

To further elaborate upen this nrocedure, suitable divalent nkers include, by way of example only, those derived from dicarboxylic acids, disulfonylhalides, dialdehydes, dipseudohalides, diketones, dihalides, diisocyanates, diamines, diols, diboronates, mixtures of carboxylic acids, sulfonylhalides, aldehydes, ketones, halides, isocyanates, amines and diols. In each case, the carboxylic acid, sulfonylhalide, aldehyde, ketone, halide, isocyanate, amine and diol functional group is reacted with a complementary functionality on the ligand to form a ) covalent linkage. Such complementary functionality is well known in the art as illustrated in the following table, which is exemplary only:

COMPLEMENTARY BINDING CHEMISTRIES

First R ive G S LR ve G Lil hydroxyl isocyanate urethane amine epoxide 3-hydroxyarmine sulfonyl halide amine sulfonamide carboxyl acid amine amide hydroxy! alkyl/aryl halide ether aldehyde amine/NaCNBH; amine ketone amine/NaCNBH; amine amine isocyanate urea

Exemplary linkers include the following linkers identified as X-1 throu gh

X-418 as set forth below ina Table 1:

po i ; i ° on T o 1 oon M ow . © =

Lone . Ag tr® : o . or Ng PVRS NP2 O jad on © oy jad on . , . ° 1 ! on, - wo Se } 2 2 o> + 3 i 7) o an o oon | “ : o oo { = on o, Oo A

ENN NPN NN LAL > | 07 Ho | 5 orig ~ 1B poe bo em,

J . % J 1} ni [t) v . a oH [] o | o om on on

A ~~ BN a adi ¢ woah, vo - | 250 ; oe ne" i i ! °o H Ke

El 14 1s! ® — " - 0)

H . 0, jo bad ” I yw i ) whom oJ) { . L)=o oP oy | AA came oo ow

A i b | A ° | wo; Re . » 20) ni o a a: © 3 ry o - Ce 0 1 00 MS

I SSE b Pr

CA Re ed EG LOSE,

Ngo L- 1 kJ — 24) o = uj . — zl - » ~ — » ° o i W ,0 X { om LA

BEN ahd H 1p =] \ Sat ol gr ° on |= 14 - i | ne : on .

N dH . 33] S » ni ——— n) _ 3) ; ON p= PRR 3 d > -° HY HE Na oY ~~ OTe EE Se SIR Re “y - To ed Tw OO - » ~ 38 oe wl o ©} wa a a

TTT TT em o rr) LOM CY nwo. .0 ° - Hp —0n c N ps ©)

Anh, P: Po vo, Pj | Hae ISS had | vo. J HE “se

Moy tr . a “| wi oo LL MR. heal | ’ ae 2 Pom ~\ = on - = — 0 =

To Tn XT | Ay TE

Lr me \ ! : — om ; Hd |e TR TE ol © © 30 pi) = . _» . 34

BUNS 4 | BUS USE Oe om | r—o ; © - on r= bo Se Ie > LL j 0 ! Cir oe | — ! ol _ — ! g) n — 7 = 2 EN L 7 ON

MALE CE | on ~~ M ~ HS aN : {Le . 12

RE + X wg o | AA Ha Po. on

W sr Err mcf On ( JA TONAL Ene” NT on Ce

Ie o | ° | oe x oe } | a n ED) 31 1 7

- .

Wr O 99/64042 PCT/US99/12728 > -— — ~ - i ° on K © hd Te BSL NEN QL. A hat wo Lo Nem - Rs Y ot id ° Se "oo EN

LY XX Tae | x » oe [2 (1) 0) 2H 1 u v | fond n os " Hat LOL \

I a CL R J f STING on pe 0, Om { . 'S yn a i ap - “oe JOR eo ; “ro To

Y) * 1 [3 LH 2 « 4 °e I hy H 2 © ° ~~ a

Ie Oo pe) . oo 0 NES | ween oN Ne SLA on

AAA yy o 4 \ ~ ; CC memo® "0. ° 3 H er qs Y [] oo nl 0) 93; 9%

O° wp rr 6F 4F 8 no necoy be H Pro 2a 5 ] Ps : a oO = v i on

APN BU SE LVN i Ee PEERY vss PP PF OF O ( 5 H] o a, he ps . ‘ 0 \ < ” 98 ™ ° 100! LH . 103 0 H rr FF FF FV ° on ; o | oc H _

SS I I EL

I a] o~X, CC: : = ~ = © ie ° i ° Ay [I ENG ee © . | Balad 2 - ° © | wom 09 110 we -— na in - "a on -— had bd i oe 2] . CY ] o ° Ce x | LS : 0 Se eo” 0

He i x yO) iss wo={ | Dom or C= ot o> on s v, . P.

Jo > 1s * nd ° un Hei ue 126 [1 oo ° Q Rad - — N Ae ] a P ) ne ee ) Fl 4 \ oN NN he ~ 2, - » ~ ° o J hd z a — ( 8 = . ° A] : “ wn! ° 132) = wi 124: 123 — 126 ——— H 2) on i 4 i h » To : ro voi o ~ . pos BOSS Ter, | SST Gha ! nf pa en

Ste NAery™ ° ! WA H G7 - oy

IEA i \ ore L oe [21] 13 asi 10! [1 132 i i

Disulfs I ! i i ee : 1 ~. ° 9 ° CS g 1,9 $a a | A : A

J ne 3 ) : o o ° Aen m [11] 138 © OL 134] - 197 = 1) - . = a " of be i Lory gre Se o ; Fo

Ss ® =. 23K _ > Py k Ce ~ re Lp §%%

Se. 19] re | w= Sr af y 2! RS red

EE a co Q, C .- _L-0

S50 "i 0 Teo [ o 2 “No pve ost 8 5 ° SY 1 © Ra PS H { a

CA B~y | " oh _ | ° WY ao @ ' v a ¢ RAN © : EER or ol 14 & Ll 148! 149 I) o, A Ao os x R . 2 5 FE ! : 0’ a ° : i m ;

— .

Dialdchydes : i — ! i on . Lg { > 7 3 “5 ! Tae we

TUT, | & C NENG NZ I I Swe be : i ° ° : ° “ - an Can i ie LN ) 19! 154 [111 [EIN] 3} 0s ~° i ' Pe 0 ~~ ° be No

H F i} PNT . 2 p B

No bg C0 oon I NI ; = A ? LR S : ' A Ra © ‘ ' . . [-] . a) : ne ‘oy 1990 _ [IH roa Ls 10. 164 # o [ HEN . =c vo= =v

On A | aC | i ! it = he “oo ! i SI A TE . a .

H i an i ! . 1851 i. see! 367) 163: 160° 310) “eo | oF" So ; [ol £30 . | ry : 2 Ae : n * i i | ' : t | :

SN - ! i . ? wv nt 172, 1! » viel : t : . I | . ! . « . | . .

Dibalides i : : : gr Beg i g “5 [RE AN MT ee 1 of a . ! . o i ood ir a Ay A ; on o% I ! :

Vas 176) 1 113 ~ Lal 130]

CNN TS | I 0 aa a RN Are TN a" foud LT wl au ws! rl as’ ee

ANSE | 0 [ -_— [7 H CNS ES ENE ey ~ rr —O I~ i. So

NATTA PAA

° 1. wl 9; 190 wl SL [A a H ° ad i on | mom rr

HOC Po) | Hr [a

Pe “ . NN .

H t oy 19! od 1981 196: 197 9 [ae Va ah I [4 [3 1 ~~ Jo ' | nC o | oy | oo on -

Rani te 199) 200 3010 202} 203. 208

AA Ld ne® ~ ° ! PEN Taam, oe wor el -~ | [9 , \ . 281 ul 201 oo ™ 209 Ed [NNN Ld Pd ; $ he po ; o Pp) | : : PEN : H : 4 sm A

H . i ° H wm’ m an! a me — : | I

H

Co

Diisocyanates i ; ; : i :

H H H i 3

. ”

Cl o, o J ' 1} OQwmzi

NS TP Zio 2 ry 4 [IE — : “ v Xs | J SURE. : Ve [) - w’ a avs o . Fo 1 a

K : Eg | | = ! Heo oy * H pS us| ne ay nel Ne 20: 0 i ”, 9 2 : ZN t PEN rN £3 4 ° ° t : «

Ar Lo XN % dN A er ~~ ye EE ae HO ee 0 © na in wd i ne en . = < i wl 223 } pod LN, 33) 32)

H 7 ~ Oy i ° al - 5 . ni oY , : pay CS ~~ [ae len &° i ° a oy : A te : Fo -.. rd : i i —.

IY 1. wn} uy’ pi) om ~~ ° Cy A nS Cama ENE hE 2 Lom py hg Seve REO A : o ay i HPA " { t : HE AS : SUN i me J Fo : No : v3 na 8. neon vi! ¢ nr ne co 3 2° N iG x AY J ~~ ! SUN iL oem i PEN ! o” H " Treg NN

H J - o EY 3 oe” LE »

A vy 10 wi 0 NE 9% 03°

Y L) .Q o = te [+] \ \, oF ] fo rN iS " on NTN 1 a's Ce ITU I a . peed ay | 2 ding i ey © EE Na wae ' H ’ ° . _ 144 us pL J8%; dat

Diamipes ; ! { : ~ H —~ Ea)

Iw - EA a IE WATT amin AD a nL) vA oy : on . (4 ! : . ' N —

EO 21i 143 wWAAAL ag

B . ~ ECE Pan Lo ~

AANA | s@)s REACHING A SW aonb a NF . : EY "~

L 1 . i ml 150 an, 330 239 0

I ES SE SE—" T

LR a a a NASH : arsed EE =

I & P) : - Ng t LL

Sah ; “n ! [a ! . 281 13 2° 160 1051 106

AD = a] LENNY ANOS ENN RVs

Core Y eB X a A on

By H ! 207 uy 29 wn: ml in

No : ~~

NN ¢ Lea NL NP WN : ~ OFC H da i Al : ba ] ha TI N oy oy NA SA » on ~ ¢ HERE oy . Sen : ~ | : “0 I mn R wal nl i m| ml

RS Oye LL Jal a Fyed \ — | A, : — POH {rm NO pe PN fms LJ DNS od Hn :

H od - : wl ol Mop N= \=/" Su. 83 m4

Oa gO WE SS ed , oy HHO om ! H HR AN YT ! i : A 2 .

H 1 m 16 ni "al 19! 20)

* - a NR Ta Po a LUNN f FE) . PAN ¢ i. Oe "oy N i em oT

Bene TE = uy La) m) wi 24! 208 29= 4 H NN LIL) WS i | ww Te SOR

Fegan) OTD o JQ, | TT 1 ; = ww faa i 2%) m I” 300) pes wm 4 NNN) =

AY ASEAN aod Me Oey = Hy yey [RA Wy 4 ; or f A wé - H Cr y ee Ne ! : H Ngee 203] 304 ol 108] wo 108

ARAN - w OOF CXC SPN; TE eo =/ ¢ y’ | ®e ay oy oy ! Ne’ Tae pM wi LY oy . oy X

AV aNd . i : wy _ pL ne ml ny 33 Ie

J a a Ta | Lon . O R | Wow = ; FN - “ “.

LL " RNC y | pl RE” oy < (. | ; we & ! Noy : 3131 tel nr ae ns )20

LN a I NT ; a . i a Nd] i : : t i 1 i 531! 23) nw, as: 3s ! : om : 1 § -_ ay, 1 Y | —— N [2 PRNEN es . a. | . o i © ! a — 0 ED

LAN r’ . ° or . [A on . ro. H Ag Aar® | A ti ¢ i

UNE ha) ! A. ] C i N, > > »zel ca) ae La 19) 1m PY] ~~ i! N ~

IYI ~ 11 i T Se Ce on Heo 0" oy AA neo : Sey Ct an o LOND of i «oN i om ! or 12 m 334 ns: pH ny a Img Og, |p TT 0 .. oo | I) : Ie oo o vs . ine Son 5, - PANN

RIE C o nel 0 Fay . o Jr ~~ oy, . oy [-Y es

La on, 1 38 339) 10 30 a) 302 103 o 03 oo 8

Pa AZ ae a a Ya | hl Po om vo on oN No i ne NS + on . 344 343) 34 347 poy bao wo INNO Ly wo ~~ i Gif [¥d - : vor See a ro. : | VA HER ~o = Ha Tr ~~?

H rr wo 239) 331 )2 In| 34, 33 oo ANN oN AYO ou or NAN 0 on | ” | Ao a

Zk A i We” "a ! Pan Oo & | ~ .

He ig | | on : x et 336) 37, ya! 359i 340! nm

I ae NES ae - af or ! AN on . id . o- : 5 EYL ; ro ly se Og w. J oy ! : . : . 32! 3%) el 48! 38 2

. .

Sa os Ne TUT C= T= o, = i ! é wo” 7 He oy 17 [dre o_J~ | ? PNA o8 : 268 39 mn nl mi < in i a rr [ee Fe ~ { hr SC SI PRET ay 9% [CP i ro Ton wo i oe 374 373 30 Rall ms! wm) ote =— wo tg or Cm gH oo oT Non on o- oT 00) 388 In 8) 1! mn

Dithiols : ee aa 3 - [J Po. ~

NNN ad at I en ! 2 i

A TR is - PN : : bes : 388] 387] J 389. 190: 9: ° a] -e. Loan oy a | [" © = Non oo i

Lean Gh Bos i > | bod Ut o - wr

Am mn 39 ™ oY wi om ° | vs RY

Bg On At Oa - ! aN LT

Ld oa re - “Yr : 3 oan ~ [

I» b} «0 fe “i 3 ©)

ANN i : ro, o Ee Ne ll I Te a TT

PRS CE : i ; : ] 04 448) 406 07] 408 ed on * on on | Bg Ire a Bs

H [] 7 H “ aaa LK, Tm ! | Re! I N

BB - - © H ale 410 414 2 a} 44 as -— r= . = EeNegirs=y

NN i eZ re - : "0 i 418! 417 418!

Representative ligands for use in this invention inc lude, by way of example, L-1 through L-11 as identified above.

Combinations of ligands (L) and linkers (X) per this invention include, by way example only, homo- and hetero-dimers wherein a first ligand is selected from

L~1 through L-11 above and the second ligand and linker is selected from the following:

L-1/X-1- L-1/X-2- L-1/X-3- L-1/X-4- L-1/X-5- L-1/X-6-

L- 1/X-7- L-1/X-8- L-1/X-9- L-1/X-10- L.-1/X-11- L-1/X-12-

L- 1/X-13- L-1/X-14- L-1/X-15- L-1/X-16- L-1/X-17- L-1/X-18-

L- 1/X-19- L-1/X-20- L-1/X-21- L-1/X-22- L-1/X-23- L-1/X-24-

L- 1/X-25- L-1/X-26- L-1/X-27- L-1/X-28- L-1/X-29- L-1/X-30-

L- 1/X-31- L-1/X-32- L-1/X-33- L-1/X-34- L -1/X-35- L-1/X-36-

L- 1/X-37- L-1/X-38- L-1/X-39- L-1/X-40- L -1/X-41- L-1/X-42-

L- 1/X-43- L-1/X-44- L-1/X-45- L-1/X-46- L-1/X-47- L-1/X-48-

L- 1/X-49- L-1/X-50- L-1/X-51- L-1/X-52- L -1/X-53- L-1/X-54-

L- 1/X-55- L-1/X-56- L-1/X-57- L-1/X-58- I.-1/X-59- L-1/X-60-

L- 1/X-61- L-1/X-62- L-1/X-63- L-1/X-64- L—1/X-65- L-1/X-66-

L-1/X-67- L-1/X-68- L-1/X-69- L-1/X-70- L-1/X-71- L-1/X-72-

L-1/X-73- L-1/X-74- L-1/X-75- L-1/X-76- L-1/X-77- L-1/X-78-

L-1/X-79- L-1/X-80- L-1/X-81- L-1/X-82- L—1/X-83- L-1/X-84-

L-1/X-85- L-1/X-86- L-1/X-87- L-1/X-88- L-1/X-89- L-1/X-90-

L-1/X-91- L-1/X-92- L-1/X-93- L-1/X-94- L-1/X-95- L-1/X-96-

L-1/X-97- L-1/X-98- L-1/X-99- L-1/X-100- L-1/X-101- L-1/X-102-

L-3/X-103- L-1/X-104- L-1/X-105- L-1/X-106- L-1/X-107- L-1/X-108-

L-1/X-109- L-1/X-110- L-1/X-111- L-1/X-112- 1-1/X-113- L-1/¥-114-

L-Z/X-115- L-1/X-116- L-1/X-117- L-1/X-118- L-1/X-119- L-1/X-120-

L-R/X-121- L-1/X-122- L-1/X-123- L-1/X-124- L-1/X-125- L-1/X-126-

L-E/X-127- L-1/X-128- L-1/X-129- L-1/X-130- L-1/X-131- L-1/X-132-

L-X /X-133- L-1/X-134- L-1/X-135- L-1/X-136- L-1/X-137- L-1/X-138-

L-1/X-139- L-1/X-140- L-1/X-141- L-1/X-142- L—1/X-143- L-1/X-144-

L-1 /X-145- L-1/X-146- L-1/X-147- L-1/X-148- L—1/X-149- L-1/X-150-

L-1/X-151- L-1/X-152- L-1/X-153- L-1/X-154- L—1/X-155- L-1/X-156-

L-1/X-157- L-1/X-158- L-1/X-159- L-1/X-160- L-1/X-161- L-1/X-162- .

L-1 /X-163- L-1/X-164- L-1/X-165- L-1/X-166- L-1/X-167- L-1/X-168-

L-1 /X-169- L-1/X-170- L-1/X-171- L-1/X-172-

L-1 /X-173- L-1/X-174- L-1/X-175- L-1/X-176- L-1/X-177- L-1/X-178-

L-1 /X-179- L-1/X-180- L-1/X-181- L-1/X-182- L-1/X-183- L-1/X-184-

L-1 /X-18S- L-1/X-186- L-1/X-187- L-1/X-188- L-1/X-189- L-1/X-190-

L-1/X-191- L-1/X-192- L-1/X-193- L-1/X-194- L-1/X-195- L-1/X-196- 40 L-1 /X-197- L-1/X-198- L-1/X-199- L-1/X-200- L-1/X-201- L-1/X-202-

L-1 /X-203- L-1/X-204- L-1/X-205- L-1/X-206- L- 1/X-207- L-1/X-208-

L-1 /X-209- L-1/X-210- L-1/X-211- L-1/X-212- L- 1/X-213- L-1/X-214-

L-1 /X-215- L-1/X-216- L-1/X-217- L-1/X-218- L- 1/X-219- L-1/X-220-

L-1 /X-221- L-1/X-222- L-1/X-223- L-1/X-224- L- 1/X-225- L-1/X-226

L-1/X-227- L-1/X-228- L-1/X-229- L-1/X-230- L-1/X-231- L-1/X-232-

L-1/X-233- L-1/X-234- L-1/X-235- L-1/X-236- L-1/X-237- L-1/X-238-

L-1/X-239- L-1/X-240- L-1/X-241- L-1/X-242- L-1/X-243- L-1/X-244-

L-1/X-245- L-1/X-246- L-1/X-247- L-1/X-248- L-1/X-249- L-1/X-250-

L-1/X-2Z51- L-1/X-252- L-1/X-253- L-1/X-254- L-1/X-255- L-1/X-256-

L-1/X-257- L-1/X-258- L-1/X-259- L-1/X-260- L-1/X-261- L-1/X-262-

L-1/X-2.63- L-1/X-264- L-1/X-265- L-1/X-266- L-1/X-267- L-1/X-268-

L-1/X-269- L-1/X-270- L-1/X-271- L-1/X-272- L-1/X-273- L-1/X-274-

L-1/X-275- L-1/X-276- L-1/X-277- L-1/X-278- L-1/X-279- L-1/X-280-

L-1/X-281- L-1/X-282- L-1/X-283- L-1/X-284- L-1/X-285- L-1/X-286-

L-1/X-2.87- L-1/X-288- L-1/X-289- L-1/X-290- L-1/X-291- L-1/X-292-

L-1/X-2.93- L-1/X-294- L-1/X-295- L-1/X-296- L-1/X-297- L-1/X-298-

L-1/X-2 99- L-1/X-300- L-1/X-301- L-1/X-302- L-1/X-303- L-1/X-304-

L-1/X-3 05- L-1/X-306- L-1/X-307- L-1/X-308- L-1/X-309- L-1/X-310-

L-1/X-3 11- L-1/X-312- L-1/X-313- L-1/X-314- L-1/X-315- L-1/X-316-

L-1/X-3 17- L-1/X-318- L-1/X-319- L-1/X-320- L-1/X-321- L-1/X-322-

L-1/X-3 23- L-1/X-324- L-1/X-325- L-1/X-326- L-1/X-327- L-1/X-328-

L-1/X-3 29- L-1/X-330- L-1/X-331- L-1/X-332- L-1/X-333- L-1/X-334-

L-1/X-3 35- L-1/X-336- L-1/X-337- L-1/X-338- L-1/X-339- L-1/X-340-

L-1/X-3 41- L-1/X-342- L-1/X-343- L-1/X-344- L-1/X-345- L-1/X-346-

L-1/X-3 47- L-1/X-348- L-1/X-349- L-1/X-350- L-1/X-351- L-1/X-352-

L-1/X-3 53- L-1/X-354- L-1/X-355- L-1/X-356- L-1/X-357- L-1/X-358-

L-1/X-3 59- L-1/X-360- L-1/X-361- L-1/X-362- L-1/X-363- L-1/X-364-

L-1/X-3 65- L-1/X-366- L-1/X-367- L-1/X-368- L-1/X-369- L-1/X-370-

L-1/X-3 71- L-1/X-372- L-1/X-373- L-1/X-374- L-1/X-375- L-1/X-376-

L-1/X-377- L-1/X-378- L-1/X-379- L-1/X-380- L-1/X-381- L-1/X-382-

L-1/X-3 83- L-1/X-384- L-1/X-385- L-1/X-386- L-1/X-387- L-1/X-388-

L-1/X-3 89- L-1/X-390- L-1/X-391- L-1/X-392- L-1/X-393- L-1/X-394-

L-1/X-3 95- L-1/X-396- L-1/X-397- L-1/X-398- L-1/X-399- L-1/X-400-

L-1/X-4 Oi- L-1/X-402- L-1/X-403- L-1/X-404- L-1/X-<105- 1.-1/X-406-

L-1/X-4-07- L-1/X-408- L-1/X-409- L-1/X-410- L-1/X—=<411- L-1/X-412-

L-1/X-4-13- L-1/X-414- L-1/X-415- L-1/X-416- L-1/X-417- L-1/X-418-

L-2/X-1 - L-2/X-2- L-2/X-3- L-2/X-4- L-2/X-5- L-2/X-6-

L2X7- L-2/X-8- L-2/X-9- L-2/X-10- L-2/X- 11- L-2/X-12-

L-2/X-1 3- L-2/X-14- L-2/X-15- L-2/X-16- L-2/X- 17- L-2/X-18-

L-2/X-1 9- L-2/X-20- L-2/X-21- L-2/X-22- L-2/X-23- L-2/X-24-

L-2/X-25- L-2/X-26- L-2/X-27- L-2/X-28- L-2/X-29- L-2/X-30-

L-2/X-31- L-2/X-32- L-2/X-33- L-2/X-34- L-2/X- 35- L-2/X-36-

L-2/X-37- L-2/X-38- L-2/X-39- L-2/X-40- L-2/X-41- L-2/X-42- 40 L-2/X-&3- L-2/X-44- L-2/X-45- L-2/X-46- L-2/X-47- L-2/X-48-

L-2/X-4&9- L-2/X-50- L-2/X-51- L-2/X-52- L-2/X- 53- L-2/X-54-

L-2/X-55- L-2/X-56- L-2/X-57- L-2/X-58- L-2/X-59- L-2/X-60-

L-2/X-61- L-2/X-62- L-2/X-63- L-2/X-64- L-2/X- 65- L-2/X-66-

L-2/X-67- L-2/X-68- L-2/X-69- - L-2/X-70- L-2/X-71- L-2/X-72- 45 L-2/X-73- L-2/X-74- L-2/X-75- L-2/X-76- L-2/X-77- L-2/X-78-

L-2/X-79- L-2/X-80- L-2/X-81- L-2/X-82- L-2/X-83- L-2/X-84-

L-2/X-885- L-2/X-86- L-2/X-87- L-2/X-88- L-2/X—-89- L-2/X-90-

L-2/X-91- L-2/X-92- L-2/X-93- L-2/X-94- L-2/X-95- L-2/X-96-

L-2/X-97- L-2/X-98- L-2/X-99- L-2/X-100- L-2/X—101- L-2/X-102- 50 L-2/X-R03- L-2/X-104- L-2/X-105- L-2/X-106- L-2/X—107- L-2/X-108-

L-2/X-R09- L-2/X-110- L-2/X-111- L-2/X-112- L-2/X~113- L-2/X-114-

L-22/X-115- L-2/X-116- L-2/X-117- L-2/X-118- L-2/X-119- L-2/X-120-

L-2/X-121- L-2/X-122- L-2/X-123- L-2/X-124- L-2/X-125- L-2/X-126-

L-2/X-127- L-2/X-128- L-2/X-129- L-2/X-130- L-2/X-131- L-2/X-132-

L-2/X-133- L-2/X-134- L-2/X-135- L-2/X-136- L-2/X-137- L-2/X-138-

L-2/X-139- L-2/X-140- L-2/X-14}- L-2/X-142- L-2/X-143- L-27X-144-

L-2/X-145- L-2/X-146- L-2/X-147- L-2/X-148- L-2/X-149- L-2/X-150-

L-2/X-151- L-2/X-152- L-2/X-153- L-2/X-154- L-2/X-155- L-2/X-156-

L-2/X-157- L-2/X-158- L-2/X-159- L-2/X-160- L-2/X-161- L-2/X-162-

L-2/X-163- L-2/X-164- L-2/X-165- L-2/X-166- L-2/X-167- L-2/X-168-

L-22/X-169- L-2/X-170- L-2/X-171- L-2/X-172-

L-22/X-173- L-2/X-174- L-2/X-175- L-2/X-176- L-2/X-177- L-2/X-178-

L-2/X-179- L-2/X-180- L-2/X-181- L-2/X-182- L-2/X-183- L-2/X-184-

L-2 /X-185- L-2/X-186- L-2/X-187- L-2/X-188- L-2/X-189- L-2/X-190-

L-2 /X-191- L-2/X-192- L-2/X-193- L-2/X-194- L-2/X-195- L-2/X-196-

L-2 /X-197- L-2/X-198- L-2/X-199- L-2/X-200- L-2/X-201- L-2/X-202-

L-2/X-203- L-2/X-204- L-2/X-205- L-2/X-206- L-2/X-207- L-2/X-208-

L-2./X-209- L-2/X-210- L-2/X-211- L-2/X-212- L-2/X-213- L-2/X-214-

L-2./X-215- L-2/X-216- L-2/X-217- L-2/X-218- L-2/X-219- L-2/X-220-

L-2/X-221- L-2/X-222- L-2/X-223- L-2/X-224- L-2/X-225- L-2/X-226-

L-24X-227- L-2/X-228- L-2/X-229- L-2/X-230- 1-2/X-231- L-2/X-232-

L-2~X-233- L-2/X-234- L-2/X-235- L-2/X-236- L-2/X-237- L-2/X-238-

L-2-X-239- L-2/X-240- L-2/X-241- L-2/X-242- L-2/X-243- L-2/X-244-

L-2~X-245- L-2/X-246- L-2/X-247- L-2/X-248- L-2/X-249- L-2/X-250-

L-27X-251- L-2/X-252- L-2/X-253- L-2/X-254- L-2/X-255- L-2/X-256-

L-2/X-257- L-2/X-258- L-2/X-259- L-2/X-260- 1.-2/X-261- L-2/X-262-

L-2/X-263- L-2/X-264- L-2/X-265- L-2/X-266- 1-2/X-267- L-2/X-268-

L-2/X-269- L-2/X-270- L-2/X-271- L-2/X-272- 1.-2/X-273- L-2/X-274-

L-2rX-27s- L-2/X-276- L-2/X-277- L-2/X-278- 1.-2/X-279- L-2/X-280-

L-2/X-281- L-2/X-282- L-2/X-283- L-2/X-284- 1.-2/X-285- L-2/X-286-

L-2/X-287- L-2/X-288- L-2/X-289- L-2/X-290- 1.-2/X-291- L-2/X-292-

L-2/X-293- L-2/X-294- L-2/X-295- L-2/X-296- 1.-2/X-297- L-2/X-298-

L-2/.X-299- L-2/X-300- L-2/X-301- L-2/X-302- J.-2/X-303- L-2/X-304-

L-2/X-305- L-2/X-306- L-2/X-307- L-2/X-308- q.-2/X-309- L-2/X-310-

L-2/7X-311- L-2/X-312- L-2/X-313- L-2/X-314- Z -2/X-315- L-2/X-326-

L-2/X-317- L-2/X-318- L-2/X-319- L-2/X-320- x .-2/X-321- L-2/X-322-

L-2/X-323- L-2/X-324- L-2/X-325- L-2/X-326- Xn -2/X-327- L-2/X-328-

L-2/2X-329- L-2/X-330- L-2/X-331- L-2/X-332- KN .-2/X-333- L-2/X-334-

L-2/7X-335- L-2/X-336- L-2/X-337- L-2/X-338- X_-2/X-339- L-2/X-340-

L-2/2X-341- L-2/X-342- L-2/X-343- L-2/X-344- K.-2/X-345- L-2/X-346- 40 L-2/2X-347- L-2/X-348- L-2/X-349- L-2/X-350- E_-2/X-351- L-2/X-352-

L-2/X-353- L-2/X-354- L-2/X-355- L-2/X-356- LL -2/X-357- L-2/X-358-

L-2/2X-359- L-2/X-360- L-2/X-361- L-2/X-362- LC.-2/X-363- L-2/X-364-

L-2/2X-365- L-2/X-366- L-2/X-367- L-2/X-368- [.-2/X-369- L-2/X-370-

L-2/X-371- L-2/X-372- L-2/X-373- L-2/X-374- [.-2/X-375-- L-2/X-376- 45 L-2/2X-377- L-2/X-378- L-2/X-379- L-2/X-380- I.-2/X-381- L-2/X-382-

L-2/=X-383- L-2/X-384- L-2/X-38s- L-2/X-386- .-2/X-387- L-2/X-388-

L-2/2X-389- L-2/X-390- L-2/X-391- L-2/X-392- L-2/X-393- 1-2/X-394-

L-2/3X-395- L-2/X-396- L-2/X-397- L-2/X-398- L .-2/X-399- L-2/X-400-

L-2/.401- L-2/X-402- L-2/X-403- L-2/X-404- L_-2/X405- L-2/X-406- 50 L-2/3.407- L-2/X-408- L-2/X-409- L-2/X-410- L-2/X-411- L-2/X-412-

L-2/-413- L-2/X-414- L-2/X-415- L-2/X-416- L-2/X417- 1-2/X-418-

L-3/X-1- L-3/X-2- L-3/X-3- L-3/X-4- L-3/X-5- L-3/X-6-

L-3/X-7- L-3/X-8- L-3/X-9- L-3/X-10- L-3/X~ 11- L-3/X-12-

L-3/X-13- L-3/X-14- L-3/X-15- L-3/X-16- L-3/X- 17- L-3/X-18-

L-3/X-19- L-3/X-20- L-3/X-21- L-3/X-22- L-3/X-~23- L-3/X-24-

L-3/X-25- L-3/X-26- L-3/X-27- L-3/X-28- L-3/X-29- L-3/X-30-

L-3/X-31- L-3/X-32- L-3/X-33- L-3/X-34- L-3/X-35- L-3/X-36- 1-3/X-37- L-3/X-38- L-3/X-39- L-3/X-40- L-3/X~41- L-3/X-42-

L-3/X-43- L-3/X-44- L-3/X-45- L-3/X-46- L-3/X~47- L-3/X-48-

L-3/X-49- L-3/X-50- L-3/X-51- L-3/X-52- L-3/X~ 53- L-3/X-54-

L-3/X-55- L-3/X-56- L-3/X-57- L-3/X-58- L-3/X~ 59- L-3/X-60-

L-3/X-61- L-3/X-62- L-3/X-63- L-3/X-64- L-3/X~65- L-3/X-66-

L-3/X-67- L-3/X-68- L-3/X-69- L-3/X-70- L-3/X~T71- L-3/X-72-

L-3/X-73- L-3/X-74- L-3/X-75- L-3/X-76- L-3/X—77- L-3/X-78-

L-3/X-79- L-3/X-80- L-3/X-81- L-3/X-82- L-3/X—83- L-3/X-84-

L-3/X-85- L-3/X-86- L-3/X-87- L-3/X-88- L-3/X~—89- L-3/X-90-

L-3/X-91- L-3/X-92- L-3/X-93- L-3/X-94- L-3/X—95- L-3/X-96-

L-3/X-97- L-3/X-98- L-3/X-99- L-3/X-100- L-3/X—101- L-3/X-102-

L-3/X-103- L-3/X-104- L-3/X-105- L-3/X-106- L-3/X—107- L-3/X-108-

L-3/X-1099- L-3/X-110- L-3/X-111- L-3/X-112- L-3/X—113- L-3/X-114-

L-3/X-115- L-3/X-116- L-3/X-117- L-3/X-118- L-3/X—119- L-3/X-120-

L-3/X-121- L-3/X-122- L-3/X-123- L-3/X-124- L-3/X—125- L-3/X-126-

L-3/X-127- L-3/X-128- L-3/X-129- L-3/X-130- L-3/X—131- L-3/X-132-

L-3/X-13%- L-3/X-134- L-3/X-135- L-3/X-136- L-3/X—137- L-3/X-138-

L-3/X-139- L-3/X-140- L-3/X-141- L-3/X-142- L-3/X—143- L-3/X-144-

L-3/X-145- L-3/X-146- L-3/X-147- L-3/X-148- 1-3/X—149- L-3/X-150-

L-3/X-1SL- L-3/X-152- L-3/X-153- L-3/X-154- L-3/X—155- L-3/X-156-

L-3/X-157- L-3/X-158- L-3/X-159- L-3/X-160- L-3/X—161- L-3/X-162-

L-3/X-163- L-3/X-164- L-3/X-165- L-3/X-166- L-3/X—167- L-3/X-168-

L-3/X-169- L-3/X-170- L-3/X-171- L-3/X-172-

L-3/X-173- L-3/X-174- L-3/X-175- L-3/X-176- L-3iX—=177- L-3/X-17%

L-3/X-179- L-3/X-180- L-3/X-181- L-3/X-182- L-3/X—183- L-3/X-184-

L-3/X-185- L-3/X-186- L-3/X-187- L-3/X-188- L-3/X-189- L-3/X-190-

L-3/X-191- L-3/X-192- L-3/X-193- L-3/X-194- L-3/X—195- L-3/X-196-

L-3/X-197- L-3/X-198- L-3/X-199- L-3/X-200- L-3/X -201- L-3/X-202-

L-3/X-203- L-3/X-204- L-3/X-205- L-3/X-206- L-3/X-207- L-3/X-208-

L-3/X-209- L-3/X-210- L-3/X-211- L-3/X-212- L-3/X-213- L-3/X-214-

L-3/X-21S- L-3/X-216- L-3/X-217- L-3/X-218- L-3/X°-219- L-3/X-220-

L-3/X-22 1- L-3/X-222- L-3/X-223- L-3/X-224- L-3/XC-225- L-3/X-226-

L-3/X-2277- L-3/X-228- L-3/X-229- L-3/X-230- L-3/X2-231- L-3/X-232- 40 L-3/X-233- L-3/X-234- L-3/X-235- L-3/X-236- L-3/X-237- L-3/X-238-

L-3/X-239- L-3/X-240- L-3/X-241- L-3/X-242- L-3/XC-243- L-3/X-244-

L-3/X-245- L-3/X-246- L-3/X-247- L-3/X-248- L-3/%C-249- L-3/X-250-

L-3/X-25 1- L-3/X-252- L-3/X-253- L-3/X-254- L-3/%&-255- L-3/X-256-

L-3/X-257- L-3/X-258- L-3/X-259- L-3/X-260- L-3/%-261. L-3/X-262- 45 L-3/X-263- L-3/X-264- L-3/X-265- L-3/X-266- L-3/><-267- L-3/X-268-

L-3/X-269- L-3/X-270- L-3/X-271- L-3/X-272- L-33€-273- L-3/X-274-

L-3/X275- L-3/X-276- L-3/X-277- 1-3/X-278- L-3/3<-279- L-3/X-280-

L-3/X-2&1- L-3/X-282- L-3/X-283- L-3/X-284- 1-3/>X-285- L-3/X-286-

L-3/X287- L-3/X-288- L-3/X-289- L-3/X-290- L-3/3<-291- L-3/X-292- 50 L-3/X293- L-3/X-294- L-3/X-295- L-3/X-296- L-3/3<-297- L-3/X-298-

L-3/X-299- L-3/X-300- L-3/X-301- L-3/X-302- L-3/3<-303- L-3/X-304-

L-3/X-305- L-3/X-306- L-3/X-307- L-3/X-308- L-3/3%-309- L-3/X-310-

L-3/X-311— L-3/X-312- L-3/X-313- L-3/X-314- L—3/X-315- L-3/X-316-

L-3/X-317—~ L-3/X-318- L-3/X-319- L-3/X-320- L—3/X-321- L-3/X-322-

L-3/X-323— L-3/X-324- L-3/X-325- L-3/X-326- L—3/X-327- L-3/X-328-

L-3/X-329— L-3/X-330- L-3/X-331- L-3/X-332- L—3/X-333- L-3/X-334- ) L-3/X-335—- L-3/X-336- L-3/X-337- L-3/X-338- L- 3/X-339- L-3/X-340-

L-3/X-341— L-3/X-342- L-3/X-343- L-3/X-344- L- 3/X-345- L-3/X-346-

L-3/X-347— L-3/X-348- L-3/X-349- L-3/X-350- L- 3/X-351- L-3/X-352-

L-3/X-353— L-3/X-354- L-3/X-355- L-3/X-356- L- 3/X-357- L-3/X-358-

L-3/X-359— L-3/X-360- L-3/X-361- L-3/X-362- L- 3/X-363- L-3/X-364-

L-3/X-365— L-3/X-366- L-3/X-367- L-3/X-368- L-_3/X-369- L-3/X-370-

L-3/X-371— L-3/X-372- L-3/X-373- L-3/X-374- L-3/X-375- L-3/X-376-

L-3/X-377- L-3/X-378- L-3/X-379- L-3/X-380- !.T3/X-381- L-3/X-382-

L-3/X-383- L-3/X-384- L-3/X-385- L-3/X-386- L-T3/X-387- L-3/X-388-

L-3/X-389- L-3/X-390- L-3/X-391- L-3/X-392- L-3/X-393- L-3/X-394- . 15 L-3/X-395- L-3/X-396- L-3/X-397- L-3/X-398- L-33/X-399- L-3/X-400-

L-3/X-401- L-3/X-402- L-3/X-403- L-3/X-404- L-33/X-405- L-3/X-406-

L.-3/X-407- L-3/X-408- L-3/X-409- L-3/X-410- L-33/X-411- L-3/X-412-

L-3/X-413- L-3/X-414- L-3/X-415- L-3/X-416- L-3/X-417- L-3/X-418-

L-4/X-1- L-4/X-2- L-4/X-3- [.-4/X-4- L-c1/X-5- L-4/X-6-

L-4/X-7- L-4/X-8- L-4/X-9- L-4/X-10- L-=/X-11- L-4/X-12-

L-4/X-13- L-4/X-14- L-4/X-15- L-4/X-16- L-21iX-17- L-4/X-18-

L-4/X-19- L-4/X-20- L-4/X-21- L-4/X-22- L-23/X-23- L-4/X-24-

L-4/X-25- L-4/X-26- L-4/X-27- L-4/X-28- L-a/X-29- L-4/X-30-

L-4/X-31- L-4/X-32- L-4/X-33- L-4/X-34- L-2/X-35- L-4/X-36-

L-4/X-37- L-4/X-38- L-4/X-39- L-4/X-40- L-M/X-41- L-4/X-42-

L-4/X-43- L-4/X-44- L-4/X-45- L-4/X-46- L-®/X-47- L-4/X-48-

L-4/X-49- L-4/X-50- L-4/X-51- L-4/X-52- L-@/X-53- L-4/X-54-

L-4/X-55- L-4/X-56- L-4/X-57- L-4/X-58- L-®/X-59- L-4/X-60-

L-4/X-61- L-4/X-62- L-4/X-63- L-4/X-64- L-®/X-65- L.-4/X-66-

L-4/X-67- L-4/X-68- L-4/X-69- L.-4/X-70- L-/X-71- L-4/X-72-

L-4/X-73- L-4/X-74- L-4/X-75- L-4/X-76- L-4=/X-77- L-4/X-78-

L-4/X-79- L-4/X-80- L4/X-81- L-4/X-82- L-45/X-83- L-4/X-84- 1-4/X-85- L-4/X-86- L-4/X-87- 1L-4/X-88- L-4-/X-89- L-4/X-90-

L-4/X-91- L-4/X-92- L-4/X-93- L-4/X-94- L-4-/X-95- L-4/X-96-

L-4/X-97- L-4/X-98- L-4/X-99- L-4/X-100- L-4-/X-101- L-4/X-102-

L-4/X-103- L-4/X-104- L-4/X-105- L-4/X-106- L-4-/X-107- L-4/X-108-

L-4/X-109- L-4/X-110- L-4/X-111- L-4/X-112- L-4-/X-113- L-4/X-114-

L-4/X-115- L-4/X-116- L-4/X-117- L-4/X-118- L-4-/X-119- L-4/X-120-

L-4/X-121- L-4/X-122- L-4/X-123- L-4/X-124- 1.-4-/X-125- L-4/X-126- 40 L-4/X-127- L-4/X-128- L-4/X-129- L-4/X-130- L-4 /X-131- L-4/X-132-

L-4/X-133- L-4/X-134- L-4/X-135- L-4/X-136- L-4./X-137- L-4/X-138-

L-4/X-139- L-4/X-140- L-4/X-141- L-4/X-142- L-4 /X-143- L-4/X-144.

L-4/X-145- L-4/X-146- L-4/X-147- L-4/X-148- L-4 jX-149- L-4/X-150-

L-4/X-151- L-4/X-152- L-4/X-153- L-4/X-154- 1-4 /X-155- L-4/X-156- 45 L-4/X-157- L-4/X-158- L-4/X-159- L-4/X-160- L-4 /X-161- L-4/X-162-

L-4/X-163- L-4/X-164- L-4/X-165- L-4/X-166- L-4./X-167- L-4/X-168-

L-4/X-169- L-4/X-170- L-4/X-171- L-4/X-172-

L-4/X-173- L-4/X-174- L-4/X-175- L-4/X-176- L-4_/X-177- L-4/X-178-

L-4/X-179- 1-4/X-180- L-4/X-181- [-4/X-182- L-4_/X-183- L-4/X-184- 50 L-4/X-185- L-4/X-186- L-4/X-187- L-4/X-188- L-4_/X-189- L-4/X-190-

L-4/X-191- L-4/X-192- L-4/X-193- L-4/X-194- L-4./X-195- L-4/X-196-

L-4/X—197- L-4/X-198- L-4/X-199- L-4/X-200- L-4/X-201- L-4/X-202-

L-4/X—203- L-4/X-204- L-4/X-205- L-4/X-206- L-4/X-207- L-4/X-208-

L-4/X—208- L-4/X-210- L-4/X-211- L-4rX-212- L-4/X-213- L-4/X-214-

L-4/X—215- L-4/X-216- L-4/X-217- L-4/X-218- L-4/X-219- L-4/X-220-

L-4/X—221- L-4/X-222- L-4/X-223- L-4/X-224- L-4/X-225- L-4/X-226-

L-4/X—227- L-4/X-228- L-4/X-229- L-4/X-230- L-4/X-231- L-4/X-232-

L-4/X—233- L-4/X-234- L-4/X-235- L-4/X-236- L-4/X-237- L-4/X-238-

L-4/X—239- L-4/X-240- L-4/X-241- L-4/X-242- L-4/X-243- L-4/X-244-

L-4/X—245- L-4/X-246- L-4/X-247- L-4/X-248- L-4/X-249- L-4/X-250-

L-4/X—-251- L-4/X-252- L-4/X-253- L-4/X-254- L-4/X-255- L-4/X-256-

L-4/X—257- L-4/X-258- L-4/X-259- L-4/X-260- L-4/X-261- L-4/X-262-

L-4/X—263- L-4/X-264- L-4/X-265- L-4/X-266- L-4/X-267- L-4/X-268-

L-4/X—269- L-4/X-270- L-4/X-271- L-4/X-272- L-4/X-273- L-4/X-274-

L-4/X—275- L-4/X-276- L-4/X-277- L-4/X-278- L-4/X-279- L-4/X-280- 15. L-4/X—281- L-4/X-282- L-4/X-283- L-4/X-284- L-4/X-285- L-4/X-286-

L-4/X—287- L-4/X-288- L-4/X-289- 1.-4/X-290- L-4/X-291- L-4/X-292-

L-4/X—293- L-4/X-294- L-4/X-295- L-4/X-296- L-4/X-297- L4/X-298-

L-4/X—299- L-4/X-300- L-4/X-301- L-4/X-302- L-4/X-303- L-4/X-304-

L-4/X—305- L-4/X-306- L-4/X-307- L-4/X-308- L-4/X-309- L-4/X-310-

L-4/X—311- L-4/X-312- L-4/X-313- L-4/X-314- L-4/X-315- L-4/X-316-

L-4/X—317- L-4/X-318- L-4/X-319- L-4/X-320- L-4/X-321- L-4/X-322-

L-4/X—323- L-4/X-324- L-4/X-325- L-4/X-326- L-4/X-327- L-4/X-328-

L-4/X—329- L-4/X-330- L-4/X-331- L-4/X-332- L-4/X-333- L-4/X-334-

L-4/X—335- L-4/X-336- L-4/X-337- L-4/X-338- L-4/X-339- L-4/X-340-

L-4/X—341- L-4/X-342- L-4/X-343- L-4/X-344- L-4/X-345- L-4/X-346-

L-4/X—347- L-4/X-348- L-4/X-349- L-4/X-350- L-4/X-351- L-4/X-352-

L-4/X—353- L-4/X-354- L-4/X-355- L-4/X-356- L-4/X-357- L-4/X-358-

L-4/X—359- L-4/X-360- L-4/X-361- L-4/X-362- L-4/X-363- L-4/X-364-

L-4/X—365- L-4/X-366- L-4/X-367- L-4/X-368- L-4/X-369- L-4/X-370-

L-4/X—371- L-4/X-372- L-4/X-373- L-4/X-374- L-4/X-375- L-4/X-376-

L-4/X—377- L-4/X-378- L-4/X-379- L-4/X-380- L-4/X-381- L-4/X-382-

L-4/X—383- L-4/X-384- L-4/X-385- L-4/X-386- 1.-4/X-387- L-4/X-388-

L-4/X—389- L-4/X-390- L4/X-391- L-4/X-392- L-4/X-393- 1-4/X-394-

L-4/X—395- L-4/X-396- L-4/X-397- L-4/X-398- 1.-4/X-399- L-4/X-400-

L-4/X—401- L-4/X-402- L-4/X-403- L-4/X-404- L-4/X-405- L-4/X-406-

L-4/X—407- L-4/X-408- L-4/X-409- L-4/X-410- L-4/X-411- L-4/X-412-

L-4/X—413- L-4/X-414- L4/X-415- L-4/X-416- L-4/X-417- L-4/X-418-

L-5/X—1- L-5/X-2- L-5/X-3- L-5/X-4- L-5/X-5- L-5/X-6- 40 L-5/X—7- L-5/X-8- L-5/X-9- L-5/X-10- L-5/X-11- . L-5/X-12-

L-5/X—13- L-5/X-14- L-5/X-15- L-5/X-16- L-5/X-17- L-5/X-18-

L-5/X—19- L-5/X-20- L-5/X-21- L-5/X-22- L-5/X-23- L-5/X-24-

L-5/X —25- L-5/X-26- L-5/X-27- L-5/X-28- L-5/X-29- L-5/X-30-

L-5X -31- L-5/X-32- L-5/X-33- L-5/X-34- L-5/X-35- L-5/X-36- 45 L-5/X -37- L-5/X-38- L-5/X-39- L-5/X-40- L-5/X-41- L-5/X-42-

L-5/X —43- L-5/X-44- L-5/X-45- L-5/X-46- L-5/X-47- L-5/X-48-

L-5/X —49- L-5/X-50- L-5/X-51- L-5/X-52- L-5/X-53- L-5/X-54-

L-5/X -55- L-5/X-56- L-5/X-57- L-5/X-58- L-5/X-59- L-5/X-60-

L-5/X -61- L-5/X-62- L-5/X-63- L-5/X-64- L-5/X-65- L-5/X-66- 50 L-5/X -67- L-5/X-68- L-5/X-69- L-5/X-70- L-5/X-71- L-5/X-72-

L-5/X-73- L-5/X-74- L-5/X-75- L-5/ZX-76- L-5/X-77- L-5/X-78-

L-5/X-79- L-5/X-80- L-5/X-81- L-5/2X-82- L-5/X-83- L-5/X-84-

L-5/X-85- L-5/X-86- L-5/X-87- L-5/2X-88- L-5/X-89- L-5/X-90-

L-5/X-91- L-5/X-92- L-5/X-93- L-5/2X-94- L-5/X-95- L-5/X-96-

L-5/X-97- L-5/X-98- L-5/X-99- L-5/2X-100- L-5/X-101- L-5/X-102-

L-5/X-103- L-5/X-104- L-5/X-105- L-5/2X-106- L-5/X-107- L-5/X-108-

L-5/X-109- L-5/X-110- L-5/X-111- L-5/X-112- L-5/X-113- L-5/X-114-

L-5/X-115- L-5/X-116- L-5/X-117- L-5/-118- L-5/X-119- L-5/X-120-

L-5/X-121- L-5/X-122- L-5/X-123- L-5/3-124- L-5/X-125- L-5/X-126-

L-5/X-127- L-5/X-128- L-5/X-129- L.-5/3-130- L-5/X-131- L-5/X-132-

L-5/X-133- L-5/X-134- L-5/X-135- L-5/3><-136- L-5/X-137- 1-5/X-138-

L-5/X-139- L-5/X-140- L-5/X-141- L-5/><-142- L-5/X-143- L-5/X-144-

L-5/X-145- L-5/X-146- L-5/X-147- L-5/3<-148- L-5/X-149- L-5/X-150-

L-5/X-151- L-5/X-152- L-5/X-153- L-5/3=<-154- L-5/X-155- L-5/X-156-

L-5/X-157- L-5/X-158- L-5/X-159- L-5/3-160- L-5/X-161- L-5/X-162-

L-5/X-163- L-5/X-164- L-5/X-165- L-5/3-166- L-5/X-167- L-5/X-168-

L-5/X-169- L-5/X-170- L-5/X-171- L-5/3«-172-

L-5/X-173- L-5/X-174- L-5/X-175- L-5/3-176- L-5/X-177- L-5/X-178-

L-5/X-179- L-5/X-180- L-5/X-181- L-5/3<-182- L-5/X-183- L-5/X-184-

L-5/X-185- L-5/X-186- L-5/X-187- L-5/3-188- L-5/X-189- L-5/X-190-

L-5/X-191- L-5/X-192- L-5/X-193- L-5/2K-194- L-5/X-195- L-5/X-196-

L-5/X-197- L-5/X-198- L-5/X-199- L-5/2Z-200- L-5/X-201- L-5/X-202-

L-5/X-203- L-5/X-204- L-5/X-205- L-5/32-206- L-5/X-207- L-5/X-208-

L-5/X-209- L-5/X-210- L-5/X-211- L-5/X2-212- L-5/X-213- L-5/X-214-

L-5/X-215- L-5/X-216- L-5/X-217- L-5/X2-218- L-5/X-219- L-5/X-220-

L-5/X-221- L-5/X-222- L-5/X-223- L-5/X2-224- L-5/X-225- L-5/X-226-

L-5/X-227- L-5/X-228- L-5/X-229- L-5/X2-230- L-5/X-231- L-5/X-232-

L-5/X-233- L-5/X-234- L-5/X-235- L-5/X2-236- L-5/X-237- L-5/X-238-

L-5/X-239- L-5/X-240- L-5/X-241- L-5/X2-242- L-5/X-243- L-5/X-244-

L-5/X-245- L-5/X-246- L-5/X-247- L-5/XZ2-248- L-5/X-249- L-5/X-250-

L-5/X-251- L-5/X-252- L-5/X-253- L-5/X2-254- L-5/X-255- L-5/X-256-

L-5/X-257- L-5/X-258- L-5/X-259- L-5/X2-260- L-5/X-261- L-5/X-262-

L-5/X-263- L-5/X-264- L-5/X-265- L-5/XZ2-266- L-5/X-267- L-5/X-268-

L-5/X-269- L-5/X-270- i-5/X-271- L-5/X-272- L-5/X-273- L-5/X-274-

L-5/X-275- L-5/X-276- L-5/X-277- L-5/X2-278- L-5/X-279- L-5/X-280-

L-5/X-281- L-5/X-282- L-5/X-283- L-5/X2-284- L-5/X-285- L-5/X-286-

L-5/X-287- L-5/X-288- L-5/X-289- L-5/XZ2-290- L-5/X-291- L-5/X-292-

L-5/X-293- L-5/X-294- L-5/X-295- L-5/XZ-296- L-5/X-297- L-5/X-298-

L-5/X-299- L-5/X-300- L-5/X-301- L-5/X-302- L-5/X-303- L-5/X-304- 40 L-5/X-305- L-5/X-306- L-5/X-307- L-5/X-308- L-5/X-309- L-5/X-310-

L-5/X-311- L-5/X-312- L-5/X-313- L-5/X2-314- L-5/X-315- L-5/X-316-

L-5/X-317- L-5/X-318- L-5/X-319- L-5/XZ-320- L-5/X-321- L-5/X-322-

L-5/X-323- L-5/X-324- L-5/X-325- L-5/X2-326- L-5/X-327- . L-5/X-328-

L-5/X-329- L-5/X-330- L-5/X-331- L-5/XZ-332- L-5/X-333- L-5/X-334- 45 L-5/X-335- L-5/X-336- L-5/X-337- L-5/X-338- L-5/X-339- L-5/X-340-

L-5/X-341- L-5/X-342- L-5/X-343- L-5/XZ-344- L-5/X-345- L-5/X-346-

L-5/X-347- L-5/X-348- L-5/X-349- L-5/XZ-350- L-5/X-351- L-5/X-352-

L-5/X-353- L-5/X-354- L-5/X-355- L-5/XC-356- L-5/X-357- L-5/X-358-

L-5/X-359- L-5/X-360- L-5/X-361- L-5/X2-362- L-5/X-363- L-5/X-364- 50 L-5/X-365- L-5/X-366- L-5/X-367- L-5/X"-368- L-5/X-369- L-5/X-370-

L-5/X-371- L-5/X-372- L-5/X-373- L-5/XZ-374- L-5/X-375- L-5/X-376-

L-5/X-377- L-5/X-378- L-5/X-379- L-5/X"-380- L-5/X-381- L-5/X-382-

L-5/X-383- L-5/X-384- L-5/X-385- L-5/X-386- L-5/X-387- L-5/X-388-

L-5/X-389- L-5/X-390- L-5/X-391- L-5/X-392- L-5/X-393- L-5/X-394-

L-5/X-395- L-5/=X-396- L-5/X-397- L-5/X-398- L-5/X-399-~ L-5/X-400-

L-5/X-401- L-5/=X-402- L-5/X-403- L-5/X-404- L-5/X-405- L-5/X-406-

L-5/X-407- L-5/2X-408- L-5/X-409- L-5/X-410- L-5/X-411- L-5/X-412-

L-5/X-413- L-5/=X-414- L-5/X-415- L-5/X-416- L-5/X-417-~ L-5/X-418-

L-6/X-1- L-6/2X-2- L-6/X-3- L-6/X-4- L-6/X-5- L-6/X-6-

L-6/X-7- L-6/2X-8- L-6/X-9- L-6/X-10- L-6/X-11- L-6/X-12-

L-6/X-13- L-6/=X-14- L-6/X-15- L-6/X-16- L-6/X-17- L-6/X-18-

L-6/X-19- L-6/2X-20- L-6/X-21- L-6/X-22- L-6/X-23- L-6/X-24-

L-6/X-25- L-6/=X-26- L-6/X-27- L-6/X-28- L-6/X-29- L-6/X-30-

L-6/X-31- L-6/=X-32- L-6/X-33- L-6/X-34- L-6/X-35- L-6/X-36-

L-6/X-37- L-6/X-38- L-6/X-39- L-6/X-40- L-6/X-41- L-6/X-42-

L-6/X-43- L-6/=X-44- L-6/X-45- L-6/X-46- L-6/X-47- L-6/X-48-

L-6/X-49- L-6/X-50- L-6/X-51- L-6/X-52- L-6/X-53- L-6/X-54-

L-6/X-55- L-6/X-56- L-6/X-57- L-6/X-58- L-6/X-59- L-6/X-60-

L-6/X-61- L-6/2%-62- L-6/X-63- L-6/X-64- L-6/X-65- L-6/X-66-

L-6/X-67- L-6/2X-68- L-6/X-69- L-6/X-70- L-6/X-71- L-6/X-72-

L-6/X-73- L-6/X-74- L-6/X-75- L-6/X-76- L-6/X-77- L-6/X-78-

L-6/X-79- L-6/2-80- L-6/X-81- L-6/X-82- L-6/X-83- L-6/X-84-

L-6/X-85- L-6/>X-86- L-6/X-87- L-6/X-88- L-6/X-89- L-6/X-90-

L-6/X-91- L-6/-92- L-6/X-93- L-6/X-94- L-6/X-95- L-6/X-96-

L-6/X-97- L-6/2X-98- L-6/X-99- L-6/X-100- L-6/X-101- L-6/X-102-

L-6/X-103- L-6/2X-104- L-6/X-105- L-6/X-106- L-6/X-107- L-6/X-108-

L-6/X-109- L-6/X-110- L-6/X-111- L-6/X-112- L-6/X-113- L-6/X-114-

L-6/X-115- L-6/X-116- L-6/X-117- L-6/X-118- L-6/X-119- L-6/X-120-

L-6/X-121- L-6/2X-122- L-6/X-123- L-6/X-124- L-6/X-125- L-6/X-126-

L-6/X-127- L-6/2X-128- L-6/X-129- L-6/X-130- L-6/X-131- L-6/X-132-

L-6/X-133- L-6/2X-134- L-6/X-135- L-6/X-136- L-6/X-137- L-6/X-138-

L-6/X-139- L-6/2X-140- L-6/X-141- L-6/X-142- L-6/X-143- L-6/X-144-

L-6/X-145- L-6/2X-146- L-6/X-147- L-6/X-148- L-6/X-149- L-6/X-150-

L-6/X-151- L-6/=X-152- L-6/X-153- L-6/X-154- L-6/X-155- L-6/X-156-

L-6/X-157- L-6/2X-158- L-6/X-159- L-6/X-160- L-6/X-161- L-6/X-162-

L-6/X-163- L-6/2-164- L-6/X-165- L-6/X-166- L-6/X-167- L-6/X-168-

L-6/X-169- L-6/2X-170- L-6/X-171- L-6/X-172-

L-6/X-173- L-6/=X-174- L-6/X-175- L-6/X-176- L-6/X-177- L-6/X-178-

L-6/X-179- L-6/=X-180- L-6/X-181- L-6/X-182- L-6/X-183- L-6/X-184-

L-6/X-185- L-6/=X-186- L-6/X-187- L-6/X-188- L-6/X-189- L-6/X-190-

L-6/X-191- L-6/2X-192- L-6/X-193- L-6/X-194- 1-6/X-195- L-6/X-196- 40 L-6/X-197- L-6/=X-198- L-6/X-199- L-6/X-200- L-6/X-201- L-6/X-202-

L-6/X-203- L-6/X-204- L-6/X-205- L-6/X-206- L-6/X-207- = L-6/X-208-

L-6/X-209- L-6/=X-210- L-6/X-211- L-6/X-212- L-6/X-213- L-6/X-214-

L-6/X-215- L-6/2X-216- L-6/X-217- L-6/X-218- L-6/X-219- L-6/X-220-

L-6/X-221- L-6/2X-222- L-6/X-223- L-6/X-224- L-6/X-225- L-6/X-226- 45 L-6/X-227- L-6/X-228- L-6/X-229- L-6/X-230- L-6/X-231- L-6/X-232-

L-6/X-233- L-6/2X-234- L-6/X-235- L-6/X-236- L-6/X-237- L-6/X-238-

L-6/X-239- L-6/=X-240- L-6/X-241- L-6/X-242- L-6/X-243— L-6/X-244-

L-6/X-245- L-6/=X-246- L-6/X-247- L-6/X-248- L-6/X-249- L-6/X-250-

L-6/X-251- L-6/2X-252- L-6/X-253- L-6/X-254- L-6/X-255- L-6/X-256- 50 L-6/X-257- L-6/2X-258- L-6/X-259- L-6/X-260- L-6/X-261- L-6/X-262-

L-6/2X-263- L-6/X-264- L-6/X-265- L-6/X-266- d.-6/X-267- L-6/X-268-

L-6/-X-269- L-6/X-270- L-6/X-271- L-6/X-272- MW _-6/X-273- L-6/X-274-

L-6/-X-275- L-6/X-276- L-6/X-277- L-6/X-278- M-6/X-279- L-6/X-280-

L-6/2X-281- L-6/X-282- L-6/X-283- L-6/X-284- M_-6/X-285- L-6/X-286-

L-6/2X-287- L-6/X-288- L-6/X-289- L-6/X-290- M_-6/X-291- L-6/X-292-

L-6/2X-293- L-6/X-294- L-6/X-295- L-6/X-296- M_-6/X-297- L-6/X-298-

L-6/=X-299- L-6/X-300- L-6/X-301- L-6/X-302- E_-6/X-303- L-6/X-304-

L-6/2X-305- L-6/X-306- L-6/X-307- L-6/X-308- K_-6/X-309- L-6/X-310-

L-6/2X-311- L-6/X-312- L-6/X-313- L-6/X-314- BH -6/X-315- L-6/X-316-

L-6/2X-317- L-6/X-318- L-6/X-319- L-6/X-320- C_-6/X-321- L-6/X-322-

L-6/2X-323- L-6/X-324- L-6/X-325- L-6/X-326- _-6/X-327- L-6/X-328-

L-6/3¢-329- L-6/X-330- L-6/X-331- L-6/X-332- I 6/X-333- L-6/X-334-

L-6/-335- L-6/X-336- L-6/X-337- L-6/X-338- I _-6/X-339- L-6/X-340-

L-6/2X-341- L-6/X-342- L-6/X-343- L-6/X-344- L_-6/X-345- L-6/X-346-

L-6/3-347- L-6/X-348- L-6/X-349- L-6/X-350- L.-6/X-351- L-6/X-352-

L-6/3-353- L-6/X-354- L-6/X-355- L-6/X-356- L_-6/X-357- L-6/X-358-

L-6/>=-359- L-6/X-360- L-6/X-361- L-6/X-362- L_-6/X-363- L-6/X-364-

L-6/>-365- L-6/X-366- L-6/X-367- L-6/X-368- L_-6/X-369- L-6/X-370-

L-6/3<-371- L-6/X-372- L-6/X-373- L-6/X-374- L_-6/X-375- L-6/X-376-

L-6/3=<-377- L-6/X-378- L-6/X-379- L-6/X-380- L_-6/X-381- L-6/X-382-

L-6/3-383- L-6/X-384- L-6/X-385- L-6/X-386- I_-6/X-387- L-6/X-388-

L-6/3«-389- L-6/X-390- L-6/X-391- L-6/X-392- I_-6/X-393- L-6/X-394-

L-6/3X-395- L-6/X-396- L-6/X-397- L-6/X-398- L_.-6/X-399- L-6/X-400-

L-6/3X-401- L-6/X-402- L-6/X-403- L-6/X-404- L_.-6/X-405- L-6/X-406-

L-6/3X-407- L-6/X-408- L-6/X-409- L-6/X-410- [_-6/X-411- L-6/X-412-

L-6/3X-413- L-6/X-414- L-6/X-415- L-6/X-416- L_-6/X-417- L-6/X-418-

L-7/3Z-1- L-7/X-2- L-7/X-3- L-7/X-4- L-7/X-5- L-7/X-6-

L-7/XZ-7- L-7/X-8- L-7/X-9- L-7/X-10- L_-7/X-11- L-7/X-12-

L-7/3&-13- L-7/X-14- L-7/X-15- L-7/X-16- L_-7/X-17- L-7/X-18-

L-7/X-19- L-7/X-20- L-7/X-21- L-7/X-22- L_-7/X-23- L-7/X-24-

L-7/X%-25- L-7/X-26- L-7/X-27- L-7/X-28- L_-7/X-29- L-7/X-30-

L-7/X-31- L-7/X-32- L-7/X-33- L-7/X-34- L_-7/X-35- L-7/X-36-

L-7/XZ-37- L-7/X-38- L-7/X-39- L-7/X-40- L_-7/X-41- L-7/X-42-

L-7/X2-43- L-7/X-44- L-7/X-45- L-7/X-46- L_-7/X-47- L-7/X-48-

L-7/X2-49- L-7/X-50- L-7/X-51- L-7/X-52- L_-7/X-53- L-7/X-54-

L-7/1XZ-55- L-7/X-56- L-7/X-57- L-7/X-58- L_-7/X-59- L-7/X-60-

L-7/X2-61- L-7/X-62- L-7/X-63- L-7/X-64- L_-7/X-65- L-7/X-66-

L-7/X-67- L-7/X-68- L-7/X-69- L-7/X-70- L_-7/X-71- L-7/X-72-

L-7/XZ2-73- L-7/X-74- L-7/X-75- L-7/X-76- L_-7X-77- L-7/X-78- 40 L-7/XZ-79- L-7/X-80- L-7/X-81- L-7/X-82- L_-7/X-83- L-7/X-84-

L-7/XZ-85- L-7/X-86- L-7/X-87- L-7/X-88- L_-7/X-89- L-7/X-90-

L-7/X2-91- L-7/X-92- L-7/X-93- L-7/X-94- L_-7/X-95- + L-7/X-96-

L-7/X2-97- L-7/X-98- L-7/X-99- L-7/X-100- L.-7/X-101- L-7/X-102-

L-7/XZ-103- L-7/X-104- L-7/X-105- L-7/X-106- L.-7/X-107- L-7/X-108- 45 L-7/X_-109- L-7/X-110- L-77X-111- L-7/X-112- L.-7/X-113- L-7/X-114-

L-7/X_-115- L-7/X-116- L-7/X-117- L-7/X-118- L_-7/X-119- L-7/X-120-

L-7/X-121- L-7/X-122- L-7/X-123- L-7/X-124- L_-7/X-125- L-7/X-126-

L-7/X2-127- L-7/X-128- L-7/X-129- L-7/X-130- L.-7/X-131- L-7/X-132-

L-7/XC-133- L-7/X-134- L-7/X-135- L-7/X-136- L.-7/X-137- L-7/X-138- 50 L-7/X_-139- L-7/X-140- L-7/X-141- L-7/X-142- L.-7/X-143- L-7/X-144-

L-7/X-1455- L-7/X-146- L-7/X-147- L-7/X-148- L-7/3-149- L-7/X-150-

L-7/X-15 1- L-7/X-152- L-7/X-153- L-7/X-154- L-7/3X-155- L-7/X-156-

L-7/X-1577- L-7/X-158- L-7/X-159- L-7/X-160- ~~ L-7/3Z-161- L-7/X-162-

L-7/X-163- L-7/X-164- L-7/X-165- L-7/X-166- L-7/><-167- L-7/X-168-

L-7/X-1699- L-7/X-170- L-7/X-171- L-7/X-172-

L-7/X-173- L-7/X-174- L-7/X-175- L-7/X-176- L-7>&-177- L-7/X-178-

L-7/X-179%- L-7/X-180- L-7/X-181- L-7/X-182- L-7/><-183- L-7/X-184-

L-7/X-185- L-7/X-186- L-7/X-187- L-7/X-188- L-7/>-189- L-7/X-190-

L-7/X-1971- L-7/X-192- L-7/X-193- L-7/X-194- L-73L-195- L-7/X-196-

L-7/X-1977- L-7/X-198- L-7/X-199- L-7/X-200- L-7/3ZX-201- L-7/X-202-

L-7/X-203- L-7/X-204- L-7/X-205- L-7/X-206- L-7/>£-207- L-7/X-208-

L-7/X-209- L-7/X-210- L-7/X-211- L-7/X-212- L-7X-213- L-7/X-214-

L-7/X215S- L-7/X-216- L-7/X-217- L-7/X-218- L-7/3Z-219- L-7/X-220.

L-7/X-221- L-7/X-222- L-7/X-223- L-7/X-224- L-73X-225- L-1/X-226-

L-7/X-22"7- L-7/X-228- L-7/X-229- L-7/X-230- L-732-231- L-7/X-232- : L-7/X-233- L-7/X-234- L-7/X-235- L-7/X-236- L-7/%Z-237- L-7/X-238-

L-7/X-39- L-7/X-240- L-7/X-241- L-7/X-242- L-13<-243- L-7/X-244-

L-7/X-45- L-7/X-246- L-7/X-247- L-7/X-248- L-7/%Z-249- L-7/X-250-

L-7/X-251- L-7/X-252- L-7/X-253- L-7/X-254- L-7/%Z-255- L-7/X-256-

L-7/X-257- L-7/X-258- L-7/X-259- L-7/X-260- L-7/3=-261- L-7/X-262-

L-7/X-263- L-7/X-264- L-7/X-265- L-7/X-266- L-1/3Z-267- L-7/X-268-

L-7/X-269- L-71X-270- L-7/X-271- L-7/X-272- L-13Z-273- L-7/X-274-

L-7/X-215- L-7/X-276- L-7/X-277- L-7/X-278- L-1/3Z-279- L-7/X-280-

L-7/X-288- L-7/X-282- L-7/X-283- L-7/X-284- L-7/XZ-285- L-7/X-286-

L-7/X-287- L-7/X-288- L-7/X-289- L-7/X-290- L-7/XZ-291- L-7/X-292-

L-7/X-293- L-7/X-294- L-7/X-295- L-7/X-296- L-7/3&-297- L-7/X-298-

L-7/X-299- L-7/X-300- L-7/X-301- L-7/X-302- L-7/%Z-303- L-7/X-304-

L-7/X-305- L-7/X-306- L-7/X-307- L-7/X-308- L-7/%2-309- L-7/X-310-

L-7/X-313- L-7/X-312- L-7/X-313- L-7/X-314- L-7/%=-315- L-7/X-316-

L-7/X-317- L-7/X-318- L-7/X-319- L-7/X.320- L.7/»=2-321- L-7/X-322-

L-7/X-323- L-7/X-324- L-7/X-325- L-7/X-326- L-7/3Z-327- L-7/X-328-

L7/X-29- L-7/X-330- L-7/X-331- L-7/X-332- L-7><-333- L-7/X-334-

L-7/X-335- L-7/X-336- L-7/X-337- L-7/X-338- L-7/3<-339- L-7/X-340-

L-7/X-34 1- L-7/X-342- L-7/X-343- L-7/X-344- L-1/3-345- L-7/X-346-

L-7/X-347- L-7/X-348- L-7/X-349- L-7/X-350- L-7/><-351- L-7/X-352-

L-7/X-353- L-7/X-354- L-7/X-355- L-7/X-356- L-1/><-357- L-7/X-358-

L-7/X-359- L-7/X-360- L-7/X-361- L-7/X-362- L-7/><-363- L-7/X-364-

L-7/X-365- L-7/X-366- L-7/X-367- L-7/X-368- L-7/3<-369- L-7/X-370-

L-7/X-371- L-7/X-372- L-7/X-373- L-7/X-374- L-7/><Z-375- L-7/X-376- 40 L-7/X-3777- L-7/X-378- L-7/X-379- L-7/X-380- L-7/3<-381- L-7/X-382-

L-7/X-383- L-7/X-384- L-7/X-385- L-7/X-386- L-7/><-387- L-7/X-388-

L-7/X-389- L-7/X-390- L-7/X-391- L-7/X-392- L-7/3<-393- L-7/X-394-

L-7/X-395- L-7/X-396- L-7/X-397- L-7/X-398- L-7/3<-399- L-7/X-400-

L-7/X401- L-7/X-402- L-7/X-403- L-7/X-404- L-7/3=<-405- L-7/X-406- 45 L-7/X-40"7- L-7/X-408- L-7/X-409- L-7/X-410- L-7/3=<-411- L-7/X-412-

L-7/X-413- L-7/X-414- L-7/X-415- L-7/X-416- L-7/3<-417- L-7/X-418-

L-8/X-1- L-8/X-2- L-8/X-3- L-8/X-4- L-8/3<-5- L-8/X-6-

L-8/X-7- L-8/X-8- L-8/X-9- L-8/X-10- L-8/3<-11- L-8/X-12-

L-8/X-13~ L-8/X-14- L-8/X-15- L-8/X-16- L-8/3<-17- L-8/X-18- 50 L-8/X-19~ L-8/X-20- L-8/X-21- L-8/X-22- L-8/3-23- L-8/X-24-

L-8/X-25- L-8/X-26- L-8/X-27- L-8/X-28- L-8/X-29- L-8/X-30-

L-8/X-31- L-8/X-32- L-8/X-33- L-8/X-34- L-8/X-35- 1.-8/X-36-

L-8/X-37- L-8/X-38- L-8/X-39- L-8/X-40- L-8/X41- L-8/X-42-

L-8/X-43- L-8/X-44- L-8/X-45- L-8/X-46- L-8/X-47- L-8/X-48-

L-8/X-49- L-8/X-50- L-8/X-51- L-8/X-52- L-8/X-53- . L-8/X-54-

L-8/X-55- L-8/X-56- L-8/X-57- L-8/X-58- L-8/X-59- L-8/X-60-

L-8/X-61- L-8/X-62- L-8/X-63- L-8/X-64- L-8/X-65- L-8/X-66-

L-8/X-67- L-8/X-68- L-8/X-69- L-8/X-70- L-8/X-71- L-8/X-72-

L-8/X-73- L-8/X-74- L-8/X-75- L-8/X-76- L-8/X-77- L-8/X-78-

L-8/X-79- L-8/X-80- 1.-8/X-81- L 8/X 82- L-8/X-83- L-8/X-84-

L-8/X-85- L-8/X-86- L-8/X-87- L-8/X-88- L-8/X-89- L-8/X-90-

L-8/X-91- L-8/X-92- L-8/X-93- L-8/X-94- L-8/X-95- L-8/X-96-

L-8/X-97- L-8/X-98- L-8/X-99- L-8/X-100- L-8/X-101- L-8/X-102-

L-8/X-103- L-8/X-104- L-8/X-105- L-8/X-106- L-8/X-107- L-8/X-108- a5 L-8/X-109- L-8/X-110- L-8/X-111- L-8/X-112- L-8/X-113- L-8/X-114-

L-8/X-115- L-8/X-116- L-8/X-117- L-8/X-118- L-8/X-119- L-8/X-120-

L-8/X-121- L-8/X-122- L-8/X-123- L-8/X-124- L-8/X-125- L-8/X-126-

L-8/X-127- L-8/X-128- L-8/X-129- L-8/X-130- L-8/X-131- L-8/X-132-

L-8/X-133- L-8/X-134- L-8/X-135- L-8/X-136- L-8/X-137- L-8/X-138-

L-8/X-139- L-8/X-140- L-8/X-141- L-8/X-142- L-8/X-143- L-8/X-144-

L-8/X-145.- L-8/X-146- L-8/X-147- L-8/X-148- L-8/X-149- L-8/X-150-

L-8/X-151- L-8/X-152- L-8/X-153- L-8/X-154- L-8/X-155- L-8/X-156-

L-8/X-157- L-8/X-158- L-8/X-159- L-8/X-160- 1.-8/X-161- L-8/X-162-

L-8/X-163- L-8/X-164- L-8/X-165- L-8/X-166- L-8/X-167- L-8/X-168-

L-8/X-169- L-8/X-170- L-8/X-171- L-8/X-172-

L-8/X-173- L-8/X-174- L-8/X-175- L-8/X-176- L-8/X-177- L-8/X-178- . L-8/X-179- L-8/X-180- L-8/X-181- L-8/X-182- L-8/X-183- L-8/X-184-

L-8/X-185- L-8/X-186- L-8/X-187- L-8/X-188- L-8/X-189- L-8/X-190-

L-8/X-191- L-8/X-192- L-8/X-193- 1-8/X-194- L.-8/X-195- L-8/X-196-

L-8/X-197- L-8/X-198- L-8/X-199- 1.-8/X-200- L-8/X-201- L-8/X-202-

L-8/X-203- L-8/X-204- L-8/X-205- 1.-8/X-206- L-8/X-207- L-8/X-208-

L-8/X-208- L-8/X-210- L-8/X-211- 1.-8/X-212- L-8/X-213- L-8/X-214-

L-8/X-215- L-8/X-216- L-8/X-217- 1.-8/X-218- L-8/X-219- L-8/X-220-

L-8/X-221- -8/X-222- L-8/X-223- 1.-8/X-224- L-8/X-225- L-8/X-226-

L-8/X-227- L-8/X-228- L-8/X-229- 1.-8/X-230- L-8/X-231- L-8/X-232-

L-8/X-233- L-8/X-234- L-8/X-235- 1.-8/X-236- L-8/X-237- L-8/X-238-

L-8/X-235- L-8/X-240- L-8/X-241- 1.-8/X-242- L-8/X-243- L-8/X-244-

L-8/X-245- L-8/X-246- L-8/X-247- A -8/X-248- L-8/X-249- L-8/X-250-

L-8/X-251- L-8/X-252- L-8/X-253- X.-8/X-254- L-8/X-255- L-8/X-256- 40 L-8/X-257- L-8/X-258- L-8/X-259- L-8/X-260- L-8/X-261- L-8/X-262-

L-8/X-263- L-8/X-264- L-8/X-265- KL -8/X-266- L-8/X-267- L-8/X-268-

L-8/X-269- L-8/X-270- L-8/X-271- E-8/X-272- L-8/X-273- L-8/X-274-

L-8/X-275- L-8/X-276- L-8/X-277- N.-8/X-278- L-8/X-279- ~~ L-8/X-280-

L-8/X-281- L-8/X-282- L-8/X-283- K.-8/X-284- L-8/X-285- L-8/X-286- 45 L-8/X-287- L-8/X-288- L-8/X-289- K.-8/X-250- L-8/X-291- L-8/X-292- [-8/X-293- L-8/X-294- L-8/X-29s- E_-8/X-296- L-8/X-297- L-8/X-298-

L-8/X-299- L-8/X-300- L-8/X-301- K.-8/X-302- L-8/X-303- L-8/X-304-

L-8/X-305- L-8/X-306- L-8/X-307- E.-8/X-308- L-8/X-309- L-8/X-310-

L-8/X-311- L-8/X-312- L-8/X-313- L.-8/X-314- L-8/X-315- L-8/X-316-

SO L-8/X-317- L-8/X-318- L-8/X-319- L_-8/X-320- L-8/X-321- L-8/X-322-

L-8/X-323- L-8/X-324- L-8/X-325- L_-8/X-326- 1-8/X-327- L-8/X-328.

L-8/X-329- L-8/X-330- L-8/X-331- E_-8/X-332- L-8/X-333- L-8/X-334.

L-8/X-335- L-8/X-336— L-8/X-337- L-8/X-338- L-8/X-339- L- 8/X-340-

L-8/X-341- L-8/X-342— L-8/X-343- L-8/X-3d4- L-8/X-345- L- 8/X-346-

L-8/X-347- L-8/X-348- L-8/X-349- L-8/X-350- L-8/X-351- L- 8/X-352-

L-8/X-353- L-8/X-354~ L-8/X-355- L-8/X-356- L-8/X-357- L- 8/X-358

L-8/X-359- L-8/X-360— L-8/X-361- L-8/X-362- L-8/X-363- L- 8/X-364-

L-8/X-365- L-8/X-366— L-8/X-367- L-8/X-368- L-8/X-369- L- 8/X-370-

L-8/X-371- L-8/X-372— L-8/X-373- L-8/X-374- L-8/X-375- L- 8/X-376-

L-8/X-377- L-8/X-378— L-8/X-379- L-8/X-380- L-8/X-381- L- 8/X-382-

L-8/X-383- L-8/X-384— L-8/X-385- L-8/X-386- L-8/X-387- L- 8/X-388.

L-8/X-389- L-8/X-390— L-8/X-391- L-8/X-392- L-8/X-393- L- 8/X-394-

L-8/X-395- L-8/X-396 - L-8/X-397- L-8/X-398- L-8/X-399- L- 8/X-400-

L-8/X-401- L-8/X-402— L-8/X-403- L-8/X-404- L-8/X-405- L- 8/X-406-

L-8/X-407- L-8/X-408 ~ L-8/X-409- L-8/X-410- L-8/X-411- L~ 8/X-412-

L-8/X-413- L-8/X-414~ L-8/X-415- L-8/X-416- L-8/X-417- L- 8/X-418-

L-9/X-1- L-9/X-2- L-9/X-3- L-9/X-4- L-9/X-5- L- 9/X-6-

L-9/X-7- L-9/X-8- L-9/X-9- L-9/X-10- L-9/X-11- L~9/X-12-

L-9/X-13- L-9/X-14- L-9/X-15- L-9/X-16- L-9/X-17- L— 9/X-18-

L-9/X-19- L-9/X-20- L-9/X-21- L-9/X-22- L-9/X-23- L—9/X-24-

L-9/X-25- L-9/X-26- L-97X-27- L-9/X-28- L-9/X-29- L—9/X-30-

L-9/X-31- L-9/X-32- L-9/X-33- L-9/X-34- L-9/X-35- L—9/X-36-

L-9/X-37- L-9/X-38- L-9/X-39- L-9/X-40- L-9/X-41- L—9/X-42-

L-9/X-43- L-9/X-44- L-9/X-45- L-9/X-46- L-9/X-47- L—9/X-48-

L-9/X-49- L-9/X-50- L-9/X-51- L-9/X-52- L-9/X-53- L—9/X-54-

L-9/X-55- L-9/X-56- L-9/X-57- L-9/X-58- L-9/X-59- L—9/X-60-

L-9/X-61- L-9/X-62- L-9/X-63- L-9/X-64- L-9/X-65- L—9/X-66-

L-9/X-67- L-9/X-68- L-9/X-69- L-9/X-70- L-9/X-71- L—9/X-T2-

L-9/X-73- L-9/X-74- L-9/X-75- L-9/X-76- L-9/X-77- L—9/X-78-

L-9/X-79- L-5/X-80- L-9/X-81- L-9/X-82- L-9/X-83- L—9/X-84.

L-9/X-85- L-9/X-86- L-9/X-87- L-9/X-88- L-9/X-89- L—9/X-90-

L-9/X-91- L-9/X-92- L-9/X-93- L-9/X-94- L-9/X-95- L—9/X-96-

L-9/X-97- L-9/X-98- L-9/X-99- L-9/X-100- L-9/X-101- L—9/X-102-

L-9/X-103- L-9/X-104-- L-9/X-105- L-9/X-106- L-9/X-107- L—9/X-108-

L-9/X-109- L-9/X-110- L-9/X-111- L-9/X-112- L-9/X-113- L—9/X-114-

L-9/X-115- L-9/X-116- L-9/X-117- L-9/X-118- L-9/X-119- L—9/X-120-

L-9/X-121- L-9/X-122- L-9/X-123- L-9/X-124- L-9/X-125- L -9/X-126-

L-9/X-127- L-9/X-128- L-9/X-129- L-9/X-130- L-9/X-131- L -9/X-132-

L-9/X-133- L-9/X-1348- L-9/X-135- L-9/X-136- L-9/X-137- L -9/X-138-

L-9/X-139- L-9/X-140- L-9/X-141- L-9/X-142- L-9/X-143- L -9/X-144-

L-9/X-145- L-9/X-146- L-9/X-147- L-9/X-148- L-9/X-149- L-9/X-150- 40 L-9/X-151- L-9/X-152- L-9/X-153- L-9/X-154- L-9/X-155- L.-9/X-156-

L-9/X-157- L-9/X-158- L-9/X-159- L-9/X-160- L-9/X-161- L--9/X-162-

L-9/X-163- L-9/X-163- L-9/X-165- L-9/X-166- L-9/X-167-° L_-9/X-168-

L-9/X-169- L-9/X-170- L-9/X-171- L-9/X-172-

L-9/X-173- L-9/X-174- L-9/X-175- L-9/X-176- L-9/X-177- L_-9/X-178- 45 L-9/X-179- L-9/X-180- L-9/X-181- L-9/X-182- L-9/X-183- L_-9/X-184-

L-9/X-185- L-9/X-186- L-9/X-187- L-9/X-188- L-9/X-189- L_-9/X-190-

L-9/X-191- L-9/X-192- L-9/X-193- L-9/X-194- L-9/X-195- L_-9/X-196-

L-9/X-197- L-9X-198- L-9/X-199- L-9/X-200- L-9/X-201- L_-9/X-202-

L-9/X-203- L-9/X-20=4- L-9/X-205- L-9/X-206- L-9/X-207- L_-9/X-208- 50 L-9/X-209- L-9/X-210- L-9/X-211- L-9/X-212- L-9/X-213- L_-9/X-214-

L-9/X-215- L-9/X-216- L-9/X-217- L—9/X-218- L-9/X-219- L-9/X-220-

L-9/X-221- L-9/X-222- L-9/X-223- L-9/X-224- L-9/X-225- L-9/X-226-

L-9/X-227- L-9/X-228- L-9/X-229- L—9/X-230- L-9/X-231- L-9/X-232-

L-9/X-233- L-9/X-234- L-9/X-235- L—-9/X-236- L-9/X-237- L-9/X-238-

L-9/X-239- L-9/X-240- L-9/X-241- L—9/X-242- L-9/X-245- L-9/X-244-

L-9/X-245- L-9/X-246- L-9/X-247- L—9/X-248- L-9/X-249- L-9/X-250-

L-9/X-251- L-9/X-252- L-9/X-253- L—-9/X-254- L-9/X-255- L-9/X-256-

L-9/X-257- L-9/X-258- L-9/X-259- L—9/X-260- L-9/X-261- L-9/X-262-

L-9/X-263- L-9/X-264- L-9/X-265- L—9/X-266- L-9/X-267- L-9/X-268-

L-9/X-269- L-9/X-270- L-9/X-271- L—9/X-272- L-9/X-273- L-9/X-274-

L-9/X-275- L-9/X-276- L-9/X-277- L—9/X-278- L-9/X-279- L-9/X-280-

L-9/X-281- L-9/X-282- L-9/X-283- L—9/X-284- -9/X-285- L-9/X-286-

L-9/X-287- L-9/X-288- L-9/X-289- L.—9/X-290- L-9/X-291- L-9/X-292-

L-9/X-293- L-9/X-294- L-9/X-295- L—9/X-296- L-9/X-297- L-9/X-298-

L-9/X-299- L-9/X-300- L-9/X-301- L-9/X-302- L-9/X-303- L-9/X-304-

L-9/X-305- L-9/X-306- L-9/X-307- L—9/X-308- L-9/X-309- L-9/X-310-

L-9/X-311- L-9/X-312- L-9/X-313- L-9/X-314- L-9/X-315- L-9/X-316-

L-9/X-317- L-9/X-318- L-9/X-319- L- 9/X-320- L-9/X-321- L-9/X-322- 1-9/X-323- L-9/X-324- L-9/X-325- L- 9/X-326- L-9/X-327- 1L-9/X-328-

L-9/X-329- L-9/X-330- L-9/X-331- L- 9/X-332. L-9/X-333- L-9/X-334-

L-9/X-335- L-9/X-336- L-9/X-337- L- 9/X-338- L-9/X-339- L-9/X-340-

L-9/X-341- L-9/X-342- L-9/X-343- L- 9/X-344- L-9/X-345- L-9/X-346-

L-9/X-347- L-9/X-348- L-9/X-349- L- 9/X-350- L-9/X-351- L-9/X-352-

L-9/X-353- L-9/X-354- L-9/X-355- L- 9/X-356- L-9/X-357- L-9/X-358-

L-9/X-359- L-9/X-360- L-9/X-361- L--9/X-362- L-9/X-363- L-9/X-364-

L-9/X-365- ~~ L-9/X-366- L-9/X-367- L-9/X-368- L-9/X-369- L-9/X-370-

L-9/X-371- L-9/X-372- L-9/X-373- L-*9/X-374- L-9/X-375- L-9/X-376-

L-9/X-377- L-9/X-378- L-9/X-379- L-=9/X-380- L-9/X-381- L-9/X-382-

L-9/X-383- L-9/X-384- L-9/X-385- L-9/X-386- L-9/X-387- L-9/X-388- 1-9/X-389- L-9/X-390- L-9/X-391- L-99/X-392- L-9/X-393- L-9/X-394-

L-9/X-395- £-9/X-396- L-9/X-397- L-29/X-398- L-9/X-399- L-9/X-400-

L-9/X-401- L-9/X-402- L-9/X-403- L-99/X-404- L-9/X-405- L-9/X-406-

L-9/X-407- L-9/X-408- L-9/X-409- L-99/X-410- L-9/X-411- L-9/X-412-

L-9/X-413- L-9/X-414- L-9/X-415- L-TIX416- L-9/X-417- L-9/X-418-

L-10/X-1- L-10/X-2- L-10/X-3- L- 10/X-4- L-107X-5- L-10/X-6-

L-10/X-7- L-10/X-8- L-10/X-9- L- 10/X-10- L-10/X-11- L-10/X-12-

L-10/X-13- L-10/X-14- L-10/X-15- L- 10/X-16- L-10/X-17- L-10/X-18-

L-10/X-19- L-10/X-20- L-10/X-21- L- 10/X-22- L-10/X-23- L-10/X-24-

L-10/X-25- L-10/X-26- L-10/X-27- L- 10/X-28- L-10/X-29- L-10/X-30- 40 L-10/X-31- L-10/X-32- L-10/X-33- L- 10/X-34- L-10/X-35- L-10/X-36-

L-10/X-37- L-10/X-38- L-10/X-39- L- 10/X-40- L-10/X-41- L-10/X-42-

L-10/X-43- L-10/X-44- L-10/X-45- L-10/X-46- L-10/X-47- L-10/X-48-

L-10/X-49- L-10/X-50- L-10/X-51- L- 10/X-52- L-10/X-53- L-10/X-54-

L-10/X-55- L-10/X-56- L-10/X-57- L- 10/X-58- L-10/X-59- L-10/X-60- 45 L-10/X-61- L-10/X-62- L-10/X-63- L-30/X-64- L-10/X-65- L-10/X-66-

L-10/X-67- L-10/X-68- L-10/X-69- L-10/X-70- L-10/X-71- L-10/X-72-

L-10/X-73- L-10/X-74- L-10/X-75- L-30/X-76- L-10/X-77- L-10/X-78-

L-10/X-79- L-10/X-80- L-10/X-81- L-130/X-82- L-10/X-83- L-10/X-84-

L-10/X-85- L-10/X-86- L-10/X-87- L-10/X-88- L-10/X-89- L-10/X-90- 50 L-10/X-91- L-10/X-92- L-10/X-93- L-130/X-94- L-10/X-95- L-10/X-96-

L-10/X-97- L-10 /X-98- L-10/X-99- L-10/X-100- L-10/X-101- L-10/X-102-

L-10/X-103- L-10 /X-104- L-10/X-105- L-10/X-106- L-10/X-107- L-10/X-108-

L-10/X-109- L-10 /X-110- L-10/X-111- L-10/X-112- L-10/X-113- L-10/X-114-

L-10/X-115- L-10 /X-116- L-10/X-117- L-10/X-118- L-10/X-119- L-10/X-120-

L-10/X-121- L-10 /X-122- L-10/X-123- L-10/X-124- L-10/X-125- L-10/X-126-

L-10/X-127- L-10/X-128- L-10/X-129- L-10/X-130- L-10/X-131- L-10/X-132-

L-10/X-133- L-10/X-134- L-10/X-135- L-10/X-136- L-10/X-137- L-10/X-138-

L-10/X-139- L-10/X-140- L-10/X-141- L-10/X-142- L-10/X-143- L-10/X-144-

L-10/X-145- L-10/X-146- L-10/X-147- L-10/X-148- L-10/X-149- L-10/X-150-

L-10/X-151- L-10/X-152- L-10/X-153- L-10/X-154- L-10/X-155- L-10/X-156-

L-10/X-157- L-10/X-158- L-10/X-159- L-10/X-160- L-10/X-161- L-10/X-162-

L-10/X-163- L-10/X-164- L-10/X-165- L-10/X-166- L-10/X-167- L-10/X-168-

L-10/X-169- L-10 /X-170- L-10/X-171- L-10/X-172-

L-10/X-173- L-10/X-174- L-10/X-175- L-10/X-176- L-10/X-177- L-10/X-178-

L-10/X-179- L-10 /X-180- L-10/X-181- L-10/X-182- L-10/X-183- L-10/X-184-

L-10/X-185- L-10/X-186- L-10/X-187- L-10/X-188- L-10/X-189- L-10/X-190-

L-10/X-191- L-10/X-192- L-10/X-193- L-10/X-194- L-10/X-195- L-10/X-196-

L-10/X-197- L-10/X-198- L-10/X-199- L-10/X-200- L-10/X-201- L-10/X-202-

L-10/X-203- L-10 /X-204- L-10/X-205- L-10/X-206- L-10/X-207- L-10/X-208-

L-10/X-209- L-10/X-210- L-10/X-211- L-10/X-212- L-10/X-213- L-10/X-214-

L-10/X-215- L-10/X-216- L-10/X-217- L-10/X-218- L-10/X-219- L-10/X-220-

L-10/X-221- L-10 /X-222- L-10/X-223- L-10/X-224- L-10/X-225- L-10/X-226-

L-10/X-227- L-10-/X-228- L-10/X-229- L-10/X-230- L-10/X-231- L-10/X-232-

L-10/X-233- L-10-/X-234- L-10/X-235- L-10/X-236- L-10/X-237- L-10/X-238-

L-10/X-239- L-10=/X-240- L-10/X-241- L-10/X-242- L-10/X-243- L-10/X-244-

L-10/X-245- L-10~/X-246- L-10/X-247- L-10/X-248- L-10/X-249- L-10/X-250-

L-10/X-251- L-10w/X-252- L-10/X-253- L-10/X-254- L-10/X-255- L-10/X-256-

L-10/X-257- L-10w/X-258- L-10/X-259- L-10/X-260- 1.-10/X-261- L-10/X-262-

L-10/X-263- L-108/X-264- L-10/X-265- L-10/X-266- L-10/X-267- L-10/X-268-

L-10/X-265- L-1G8/X-270- L-10/X-271- L-10/X-272- L-10/X-273- L-10/X-274-

L-10/X-275- L-108/X-276- L-10/X-277- L-10/X-278- L-10/X-279- L-10/X-280-

L-10/X-281- L-108/X-282- L-10/X-283- L-10/X-284- L-10/X-285- L-10/X-286-

L-10/X-287- L-108/X-288- L-10/X-289- L-10/X-290- L-10/X-291- L-10/X-292-

L-10/X-293- L-108/X-294- L-10/X-295- L-10/X-296- L-10/X-297- L-10/X-298-

L-10/X-299- L-10W/X-300- L-10/X-30)- L-10/X-302- L-10/X-303- L-10/X-304-

L-10/X-305- L-108/X-306- L-10/X-307- L-10/X-308- L-10/X-309- ° L-10/X-310-

L-10/X-311- L-1G¥/X-312- L-10/X-313- L-10/X-314- L-10/X-315- L-10/X-316-

L-10/X-317- L-1O/X-318- L-10/X-319- L-10/X-320- L-10/X-321- L-10/X-322-

L-10/X-323- L-10O/X-324- L-10/X-325- L-10/X-326- L-10/X-327- L-10/X-328- 40 L-10/X-329- L-10/X-330- L-10/X-331- L-10/X-332- L-10/X-333- L-10/X-334-

L-10/X-335- L-10/X-336- L-10/X-337- L-10/X-338- L-10/X-339- L-10/X-340-

L-10/X-341- L-10/X-342- L-10/X-343- L-10/X-344- L-10/X-345- L-10/X-346-

L-10/X-347- L-10/X-348- L-10/X-349- L-10/X-350- L-10/X-351- L-10/X-352-

L-10/X-353- L-10/X-354- L-10/X-355- L-10/X-356- L-10/X-357- L-10/X-358- 45 L-10/X-359- L-10/X-360- L-10/X-361- L-10/X-362- L-10/X-363- L-10/X-364-

L-10/X-365- L-10/X-366- L-10/X-367- L-10/X-368- L-10/X-369- L-10/X-370-

L-10/X-371- L-10O/X-372- L-10/X-373- L-10/X-374- L-10/X-375- L-10/X-376—

L-10/X-377- L-10/X-378- L-10/X-379- L-10/X-380- L-10/X-381- L-10/X-382-~

L-10/X-383- L-10/X-384- L-10/X-385- L-10/X-386- L-10/X-387- L-10/X-388— 50 L-10/X-389- L-10O/X-390- L-10/X-391- L-10/X-392- L-10/X-393- L-10/X-394—

L-10/X-395- L-10/X-396- L-10/X-397- L-10/X-398- L-10/X-399- L-10/X-400—

L-10/X-401- L-10/X-402- L-10/X-403- L-10/X-404- L-10/X-405- L-10/X-406—

L-10/X-407- L-10/X-408- L-10/X-409- L-10/X-410- L-10/X411- L-10/X-412-

L-10/X413- L-10/X-414- L-10/X-415- L-10/X-416- L-10/X-417- L-10/X-418-

L-11/X-1- L-11/X-2- L-11/X-3- L-11/X-4- L-11/X-5- L-11/X-6-

L-11/X-7- L-11/X-8- L-11/X-9- L-11/X-10- L-11/X-11- L-11/X-12-

L-11/X-13- L-11/X-14- L-11/X-15- L-11/X-16- L-11/X-17- L-11/X-18-

L-11/X-19- L-11/X-20- L-11/X-21- L-11/X-22- L-11/X-23- L-11/X-24-

L-11/X-25- L-11/X-26- L-11/X-27- L-11/X-28- L-11/X-29- L-11/X-30-

L-11/X-31- L-11/X-32- L-11/X-33- L-11/X-34- L-11/X-35- L-11/X-36-

L-11/X-37- L-11/X-38- L-11/X-39- L-11/X-40- L-11/X-41- L-11/X-42-

L-11/X-43- L-11/X-44- L-11/X-45- L-11/X-46- L-11/X-47- L-11/X-48-

L-11/X-49- L-11/X-50- L-11/X-51- L-11/X-52- L-11/X-53- L-11/X-54-

L-11/X-55- L-11/X-56- L-11/X-57- L-11/X-58- L-117X-59- L-11/X-60-

L-11/X-61- L-11/X-62- L~-11/X-63- L-11/X-64- L-11/X-65- L-11/X-66-

L-11/X-67- L-11/X-68- L-11/X-69- L-11/X-70- L-11/X-71- L-11/X-72-

L-11/X-73- L-11/X-74- L-11/X-75- L-11/X-76- L-11/X-77- L-11/X-78-

L-11/X-79- L-11/X-80- L-11/X-81- L-11/X-82- L-11/X-83- L-11/X-84-

L-11/X-85- L-11/X-86- L-11/X-87- L-11/X-88- L-11/X-89- L-11/X-90-

L-11/X-91- L-11/X-92- L-11/X-93- L-11/X-94- L-11/X-95- L-11/X-96-

L-11/X-97- L-11/X-98- L- 11/X-99- L-11/X-100- L-11/X-101- L-11/X-102-

L-11/X-103- L-11/X-104- L-11/X-105- L-11/X-106- L-11/X-107- L-11/X-108-

L-11/X-109- L-11/X-110- L-11/X-111- L-11/X-112- L-1UUX-113- L-11/X-114-

L-11/X-115- L-11/X-116- L-11/X-117- L-11/X-118- L-11/X-119- L-11/X-120-

L-11/X-121- L-11/X-122- L-11/X-123- L-11/X-124- L-11/X-125- L-11/X-126-

L-11/X-127- L-11/X-128- L- 11/X-129- L-11/X-130- L-11/X-131- L-11/X-132-

L-11/X-133- L-11/X-134- L- 11/X-135- L-11/X-136- L-1UX-137- L-11/X-138-

L-11/X-139- L-11/X-140- L- 11/X-141- L-11/X-142- L-1U/X-143- L-11/X-144-

L-11/X-145- L.-11/X-146- L- 11/X-147- L-11/X-148- L-11/X-149- L-11/X-150-

L-11/X-151- L-11/X-152- L-11/X-153- L-11/X-154- L-11/X-155- L-11/X-156-

L-11/X-157- L-11/X-158- L- 11/X-159- L-11/X-160- L-11/X-161- L-11/X-162-

L-11/X-163- L-11/X-164- L-11/X-165- L-11/X-166- L-11/X-167- L-11/X-168-

L-11/X-169- L-11/X-170- L-11/X-171- L-11/X-172-

L-11/X-173- L-11/X-174- L-ZU/X-175- L-11/X-176- L-11/X-177- L-11/X-178-

L-11/X-179- L-11/X-180- L-11/X-181- L-11/X-182- L-11/X-183- L-11/X-184-

L-11/X-185- L-11/X-186- L-11/X-187- L-11/X-188- L-11/X-189- L-11/X-190-

L-11/X-191- L-11/X-192- L-11/X-193- L-1I/X-194- L-11/X-195- L-11/X-196-

L-11/X-197- L-11/X-198- L-11/X-199- L-11/X-200- L-11/X-201- L-11/X-202-

L-11/X-203- L-11/X-204- L-11/X-205- L-11/X-206- L-11/X-207- L-11/X-208-

L-11/X-209- L-11/X-210- L-11/X-211- L-11/X-212- L-11/X-213- L-11/X-214-

L-11/X-215- L-11/X-216- L-11/X-217- L-11/X-218- L-11/X-219- L-11/X-220- 40 L-11/X-221- L-11/X-222- L-X1/X-223- L-11/X-224- L-11/X-225- L-11/X-226-

L-11/X-227- L-11/X-228- L-11/X-229- L-11/X-230- L-11/X-231- L-11/X-232-

L-11/X-233- L-11/X-234- L-31/X-235- L-11/X-236- L-11/X-237- L-11/X-238-

L-11/X-239- L-11/X-240- L-X1/X-241- L-11/X-242- L-11/X-243- L-11/X-244-

L-11/X-245- L-11/X-246- L-11/X-247- L-11/X-248- L-11/X-249- L-11/X-250- 45 L-11/X-251- L-11/X-252- L-X 1/X-253- L-11/X-254- L-11/X-255- L-11/X-256-

L-11/X-257- L-11/X-258- L-21/X-259- L-11/X-260- L-11/X-261- L-11/X-262-

L-11/X-263- L-11/X-264- L-11/X-265- L-11/X-266- L-11/X-267- L-11/X-268-

L-11/X-269- L-11/X-270- L-11/X-271- L-11/X-272- L-11/X-273- L-11/X-274-

L-11/X-275- L-11/X-276- L-11/X-277- L-11/X-278- L-11/X-279- L-11/X-280- 50 L-11/X-281- L-11/X-282- L-11/X-283- L-11/X-284- 1-11/X-285- L-11/X-286-

L-11/X-287- L-11/X-288- L-11/X-289- L-11/X-290- L-11/X-291- L-11/X-292-

L-11/X-293- L-11/X-294- L-11/X-295. L-11/X-296- L-11/X-297- L-11/X-298-

L-11/X-299- L-11/X-300- L-11/X-301- L-11/X-302- L-11/X-303- L-11/X-304-

L-11/X-305- L-11/X-306- L-11/X-307- L-11/X-308- L-11/X-309- L-11/X-310- } 5 L-11/X-311- L-11/X-312- L-11/X-313- L-11/X-314- L-11/X-315- L-11/X-316-

L-11/X-317- L-11/X-318- L-11/X-319- L-11/X-320- L-11/X-321- L-11/X-322-

L-11/X-323- L-11/X-324- L-11/X-325- L-11/X-326- L-11/X-327- L-11/X-328-

L-11/X-329- L-11/X-330- L-11/X-331- L-11/X-332- L-11/X-333- L-11/X-334-

L-11/X-335- L-11/X-336- L-11/X-337- L-11/X-338- L-11/X-339- L-11/X-340-

L-11/X-341- L-11/X-342- L-11/X-343- L-11/X-344- L-11/X-345- L-11/X-346-

L-11/X-347- L-11/X-348- L-11/3X-349- L-11/X-350- L-11/X-351- L-11/X-352-

L-11/X-353- L-11/X-354- L-11/X-355- L-11/X-356- L-11/X-357- L-11/X-358-

L-11/X-359- L-11/X-360- L-11/X-361- L-11/X-362- L-11/X-363- L-11/X-364-

L-11/X-365- L-11/X-366- L-11/X-367- L-11/X-368- L-11/X-369- L-11/X-370-

L-11/X-371- L-11/X-372- L-11/X-373- L-11/X-374- L-11/X-375- L-11/X-376-

L-11/X-377- L-11/X-378- L-11/X-379- L-11/X-380- L-11/X-381- L-11/X-382-

L-11/X-383- L-11/X-384- L-11/X-385- L-11/X-386- L-11/X-387- L-11/X-388-

L-11/X-389- L-11/X-390- L-11/X-39]- L-11/X-392- L-11/X-393- L-11/X-394-

L-11/X-395- L-11/X-396- L-11/X-397- L-11/X-398- L-11/X-399- L-11/X-400-

L-11/X-401- L-11/X-402- L-11/X-403- L-11/X-404- L-11/X-405- L-11/X-406-

L-11/X-407- L-11/X-408- L-11/X-409- L-11/X-410- L-11/X-411- L-11/X-412-

L-11/X-413- L-11/X-414- L-11/X-415- L-11/X-416- L-11/X-417- L-11/X-418- pi ical F lations

When employed as pharmaceuticals, the compounds of formula I are usually administered in the form of pharmaceutical compositions. These compounds can be administered by a variety of routes including oral, rectal, transdermal, subcutaneous, intravenous, intramuscular, and intranasal. These compounds are effective as both injectable and oral compositions. Such compositions are prepared in a manner well known in the pharmaceutical art and comprise at least one active compound.

This invention also includes pharmaceutical compositions which contain, as the active ingredient, one or more of the compounds of formula I above associated with pharmaceutically acceptable carriers. In making the compositions of this invention, the active ingredient is usually mixed with an excipient, diluted by an excipient or enclosed within such a carrier which can be in the form of a capsule, sachet, paper or other container. When the excipient serves as a diluent, it can be a solid, semi-solid, or liquid material, which acts as a vehicle, carrier or medium _

for the active ingredient. Thus, the compositions can be in the form of tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as a solid or in a liquid medium), ointments containing , for example, up to 10% by weight of the active compound, soft and hard gelatin capsules, suppositories, sterile injectable solutions, and steril e packaged powders.

In preparing a formulation, it may be necessary to mill the active compound to provide the appropriate particle size prior to combining with the other ingre=dients. If the active compound is substantially insoluble, it orc(Rinarily is milled to a particle size of less than 200 mesh. If the active compound is substantial 1y water soluble, the particle size is normally adjusted by millirg to provide a substantially uniform distribution in the formulation, e.g. about 40 mesh.

Sorme examples of suitable excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum acacia, calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone cellulose, sterile water, syrup, and methyl cellulose. The formulations can additionally include: lubricating agents such as talc, magnesium stearate, and mineral oil. ; wetting agents; emulsifying and suspending agents; preserving agents such as me=thyl- and propylhydroxy-benzoates; sweetening agents; and fla-voring agents. The compositions of the invention can be formulated so as to provide quick, sust ained or delayed release of the active ingredient after administr-ation to the patient by employing procedures known in the art.

The= compositions are preferably formulated in a unit dosage form._, each dosage comtaining from about 5 to about 100 mg, more usually about 10 to about mg, of tthe active ingredient. The term "unit dosage forms" refers to physically discrete units suitable as unitary dosages for human subjects and other mammals, each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect, in association wwith a suitable pharmaceutical excipient. Preferably, the compound of formula I above is employed at no more than about 20 weight percent of the pharmaceutical composition, more preferably no more than about 15 vweight percent, with the balance being pharmaceutically inert carrier(s).

The active compound is effective over a wide closage range and is generally administered in a pharmaceutically effective amount. It, will be understood, however, that the amount of the compounel actually administered will be determined by a physician or veterinarian, in the ligzht of the relevant circumstances, including the condition to be treated, thae chosen route of administration, the actual compound administered and its relative activity, the age, weight, and response of the individual patient, the severity of the patient's symptoms, and the like.

For preparing solid compositions such as table®s, the principal active ingredient is mixed with a pharmaceutical excipient to form a solid preformulation composition containing a homogeneous mixture of a compound of the present invention. When referring to these preformulation cormpositions as homogeneous, it is meant that the active ingredient is dispersed evenly throughout the composition so that the composition may be readily subdivided into equally effective unit dosage forms such as tablets, pills and capsules. This solid preformulation is then subdivided into unit dosage forrns of the type described above containing from, for example, 0.1 to about S00 mg of the active ingredient of the present invention.

The tablets or pills of the presen.t invention may be coated or otherwise compounded to provide a dosage form affording the advantage of prolonged action. For example, the tablet or pill ¢ an comprise an inner dosage and an oute=r dosage component, the latter being in th_e form of an envelope over the former.

The two components can be separated by an enteric layer which serves to resist disintegration in the stomach and permit: the inner component to pass intact into ®he duodenum or to be delayed in release. A variety of materials can be used for such enteric layers or coatings, such materials including a number of polymeric acids and mixtures of polymeric acids with su«ch materials as shellac, cetyl alcohol, anci cellulose acetate.

The liquid forms in which the novel compositions of the present inventiom may be incorporated for administration orally or by injection include aqueous solutions, suitably flavored syrups, aque=ous or oil suspensions, and flavored emulsions with edible oils such as corn Oil, cottonseed oil, sesame oil, coconut o il, or peanut oil, as well as elixirs and simiBar pharmaceutical vehicles.

Compositions for inhalation or imsufflation include solutions and suspensions in pharmaceutically acceptable, aqueous or organic solvents, or mixtures thereof, and powders. The liquid or solid compositions may contain suitable pharmaceutically acceptable excipients as described supra. Preferably tlhe compositions are administered by the orzal or nasal respiratory route for local or systemic effect. Compositions in prefer-ably pharmaceutically acceptable solvent s may be nebulized by use of inert gases. Nebulized solutions may be inhaled directly from the nebulizing device or thme nebulizing device may be attached 10 a= face mask tent, or intermittent positive poressure breathing machine. Solution, suspension, or powder compositions ma y be administered, preferably orally or nasally, from devices which deliver the formulation in an appropriate manner.

The following fosrmulation examples illustrate representative pharmaceutical compositions of the present invention.

Formulation Example 1

Hard gelatin cap=sules containing the following ingredients are pre pared:

Quantity

Ingredient {mg/capsule)

Active Ingredient 30.0

Starch 305.0

Magnesium stearate 5.0

The above ingreclients are mixed and filled into hard gelatin capsize les in 340 mg quantities. ’ Formulation Example 2

A tablet formula is prepared using the ingredients below:

Quantity

Ingredient (mg/tablet)

Active Ingredien t 25.0

Cellulose, micro<crystalline 200.0

Colloidal silicon dioxide 10.0

Stearic acid 5.0

The components are blended and compressed to form tablets, eacka weighing 240 mg.

Formulation Example 3

A dry powder in_baler formulation is prepared containing the foll> wing components:

Ingredient Weight %

Active Ingredient 5

Lactose 95

The active ingredient is mixed with the lactose and the mixture is aclded to a dry powder inh.aling appliance.

Formulation Example 4

Tablets, each containing 30 mg of active ingredient, are prepared as

S follows:

Quantity

Ingrediens (mg/tablet)

Active Ingredient 30.0 mg

Starch 45.0 mg

Microcrystalline cellulose 35.0 mg

Polyvinylgpyrrolidone (as 1.0% solution in sterile water) 4.0 mg

Sodium carboxymethyl starch 4.5 mg

Magnesiumm stearate 0.5 mg

Talc 1.0 mg

Total 120 mg

The actives ingredient, starch and cellulose are passed through a No. 20 mesh U.S. sieve and mixed thoroughly. The solution of polyvinylpyrrolidome is mixed with the resultant powders, which are then passed through a 16 mesh U.S. sieve. The granules so produced are dried at 50° to 60°C and passed through a 16 mesh U.S. sieve. The sodium carboxymethyl starch, magnesium stearate, a nd talc, previously passed through a No. 30 mesh U.S. sieve, are then added to the granules which, after mixing, are compressed on a tablet machine to yield tablets each weighing 120 mg.

Formulation Exarnple 5

Capsules, each containing 40 mg of medicament are made as follows:

Ingredient Quantity (mg/capsule)

Active Ingredient 40.0 mg

Starch 109.0 mg

Magnesium stearate : 1.0 mg

Total 150.0 mg

The active ingredient, starch, and magmesium stearate are blended, passed through a No. 20 mesh U.S. sieve, and filled irato hard gelatin capsules in 150 mg quantities.

Formulation Exarnple 6

Suppositories, each containing 25 mg of active ingredient are made as follows:

Ingredient Amount

Active Ingredient 25 mg

Saturated fatty acid glycerides to 2,000 mg

The active ingredient is passed through a No. 60 mesh U.S. sieve and suspended in the saturated fatty acid glycerides previously melted using the minimum heat necessary. The mixture is then poured into a suppository mold of nominal 2.0 g capacity and allowed to cool.

Formulation Example 7

Suspensions, each containing 50 mg of medicament per 5 .0 mL dose are made as follows:

Ingredient Amount

Active Ingredient 50.0 mg

Xanthan gum 4.0 mg

Sodium carboxymethyl cellulose (11%)

Microcrystalline cellulose (89%) 50.0 mg

Sucrose 1.75 ¢

Sodium benzoate 10.0 mg

Flavor and Color q.v.

Purified water to 5.0 mL

The active ingredient, sucrose and xanthan gum are blended, passed through a No. 10 mesh U.S. sieve, and then mixed with a previously made solution of the microcrystalline cellulose and sodium carboxymethyl cellulose in “water. The sodium benzoate, flavor, and color are dilute d with some of the water and added with stirring. Sufficient water is then added to produce the required wolume.

Formulation Example 8

Quantity

Active Ingredient 15.0 mg

Starch 407.0 mg

Magnesium stearate 30mg

Total 425.0 mg

The active ingredient, starch, and magnesium stearate are blended, passed through a No. 20 mesh U.S. sieve, and filled into hard gelatin capsules in 425.0 rng quantities.

Formulation Example 9

A formulation may be prepared as follows: 1 i

Active Ingredient 5.0 mg

Corn Oil 1.0mL

Formulation Example 10

A topical formulation may be prepared as follows:

Ingredient Quantity

Active Ingredient 1-10 g

Emulsifying Wax 30g

Liquid Paraffin 20g “White Soft Paraffin to 100 “The white soft paraffin is heated until molten. The liquid paraffin and emulsifying wax are incorporated and stirred until dissolved. Thue active ingredient is added and stirring is continued until dispersed. The mixture is then cooled until solid.

Another preferred formulation employed in the methods ©f the present invention employs transdermal delivery devices ("patches"). Such transdermal patches may be used to provide continuous or discontinuous infu sion of the compounds of the present invention in controlled amounts. The construction and use of transdermal patches for the delivery of pharmaceutical agents is well known intheart. See, e.g., U.S. Patent 5,023,252, issued June 11, 1991, herein incorporated by reference. Such patches may be constructed for continuous, pulsatile, or on demand delivery of pharmaceutical agents.

Frequently, it will be desirable or necessary to introduce the pharmaceutical composition to the brain, either directly or indirectly. Direct

Techniques usually involve placement of a drug delivery” catheter into the host's wentricular system to bypass the blood-brain barrier. ne such implantable delivery system used for the transport of biological fact ors to specific anatomical regions of the body is described in U.S. Patent 5,011,472, which is herein 1 ncorporated by reference.

Indirect techniques, which are generally preferred usually involve f-ormulating the compositions to provide for drug latenti ation by the conversion of hydrophilic drugs into lipid-soluble drugs. Latentiation is generally achieved through blocking of the hydroxy, carbonyl, sulfate, and primary amine groups present on the drug to render the drug more lipid soluble and amenable to transportation across the blood-brain barrier. Alternatively, the delivery of hydrophilic drugs may be enhanced by intra-arterial infusion of hypertonic seolutions which can transiently open the blood-brain bar rier.

Other suitable formulations for use in the presen t invention can be found in

Remington's Pharmaceutical Sciences.”

The following assays are used to evaluate the mualti-binding compounds of thais invention.

Ira vitro binding assay

Using the alpha-1-adrenoceptor binding assay de scribed by Ford et al.” te affinity (pKi) with the alpha-1A adrenergic receptor is determined for the multi-binding compounds of this invention.

Ex vivo model

Using the guinea pig prostate assay described by Haynes et al. MN prostate contractile response to the multi-binding compounds of t his invention is measured.

In vivo rmodels

Using the conscious dog model described by Hancock ext al. 12 the effects of the mrultibinding compounds on prostatic muscle response is tested.

Asdditionally, the challenge intrauretheral pressure (“cll_JP”) model can be used to rmeasure of efficacy of the multibinding compounds and the spontaneously hypertensive rat (“SHR”) model can be used to measure the sel ectivity of the multibineding compounds. Both of these models are described bsy Meyer et al. 13

Utility

T he compounds of this invention are useful for modulating alpha-1A adrenerg ic receptor activity and accordingly, may be used for tke treatment of benign prostatic hyperplasia and/or hypertension in animals, including humans.

More particularly the compounds may be used in the treatment of medical and veterinary conditions in mammals.

T he compounds of the invention are paiticularly useful En treating benign prostatic hyperplasia mediated in one form or another by alpha— 1A adrenergic receptor activity. Accordingly, the invention also relates to phzarmaceutical composition comprising a pharmaceutically acceptable excipien t and an effective amount Of a compound of the invention.

Asdditionally, the compounds of the invention may be bound to affinity resins fo r affinity chromatography. The compounds of the invesntion may be used as a tool in immunoprecipitation. The compounds may be used to identify a receptor in vitro for example in microscopy, electrophoresis and chromatography.

In order to futher illustrate the present invention and advantages themreof, the following specific examples are given but are not meant to limit the scop € of the claims in any way.

EXAMPLES

In the examples below, all temperatures are in degrees Celsius (unless otherwise indicated) and all percentages are weight percentages (also unless otherwise indicated). Further, X as used in the reactive schemes depicted in the

Examples below deno tes a halide, preferably bromide or chloride, as designated in the accompanying experimental text.

Examples 1-19 are given as representative examples of methods for preparing compounds of this invention.

In the examples below, the following abbreviations have the following= meanings. If an abbre=viation is not defined, it has its generally accepted meas=ning.

A = Angstroms cm = centimeter

DIC == 2-dimethylaminoisopropyl chloride hydrochloride

DCC = N,N-dicyclohexylcarbodiimide

DCM = dichloromethane

DIPEA = diisopropylethylamine

DMA = N, N-dimethylacetamide

DMAP == 4-N,N-dimethylaminopyridine

DMF == N, N-dimethylformamide

DMSO = dimethylsulfoxide .

DPPA == diphenylphosphory] azide g = gram

HBTU == 1-hydroxybenzotrizole

HPLC == high performance liquid chromatography

Hunig's base == diisopropylethylamine

MFC = minimum fungicidal concentration mg = milligram

MIC = minimum inhibitoxy concentration min = minute mL = milliliter mm = millimeter mmol = millimol

N = normal

PyBOP = pyridine benzotria zol-1-yloxy-tris(dimethyl- amino)phosphonitem hexafluorophosphate 1-BOC = fert-butyloxycarbonyl

TBAF = tetrabutyl ammoni um fluoride

TFA = trifluoroacetic acicd

THF = tetrahydrofuran ne =microliters

Based on the basic pharmacophore for alpha-1A adrenergic receptor antagonists and the current available materials, several classes of bivalent alpha-1A adrenergic receptor antagonists are designe d, and the syntheses are described in the following examples.

EXAMPLE 1: Preparation of a Formula | compound wherein p is 2, q is I, and the ligand, L, is the phenethylamine moiety, of Tamsulosin linked via the amino group to the linker, X.

Example 1: Tamsulosin Analogs

HNSO o NHR HN SO, SY SONH,

JT NaCNBH 3, EtOH JT OMe

Chem Pharm Bull. v 40 (6) 1992, 1443.1451. R = -CHaCH;0CHCHy- —erame ve ose Ae

R preferably contains alkyl, alkyl-ether linkages, hydroxyl grps, hydrogen bond acceptors, aryloxy, heteroaryl groups

A solution of 2,2’-oxybis(ethylamirae) (1 mmol) in methanol (4 mL) is acidified with acetic acid to pH 6.6 (pH meter) under a nitrogen atmosphere. The ketone, 1-(4-methyoxy-3-aminosulfonyl)ptwenyl-2-oxopropane (Chem. Pharm.

Bull. 1992, 40, 1443-51; 2 mmol) is added neat followed by sodium cyanoborohydride (2.1 mmol). The course of the reaction is followed by thin layer chromatography. After reaction occurs, thane reaction solution is quenched in water

) © WO 9964042 PCT/US99/12728 zaand the pH of the aqueous mixture is adjusted to 9-10 with aqueous Na, C05. The mixture is extracted with ether, the organic ex®racts are washed with half-saturated saline, dried (Na,S0,), filtered and concentrated under reduced pressure to give che crude product. The desired Formula I comapound (where R is —CH,CH,0CH,CH,-) is obtained by purification of the crude product by use of

HPLC.

EXAMPLE 2: Preparation of a Formula 1 com pound wherein p is 2, q is 1 and the

Imgand, L, is O-desethyl-tamsulosin linked via the aryloxy group to the linker, X.

T amsulosin Analog

R preferably contains alkyl, alkyt-ether linkages, hydro grps, hydrogen bond acceptors, aryloxy, heteroaryl groups

Example 2: Tamsulosin Analogs 1. NaB(CN)H,, MeOH

H,NSO, o 2. (BOC),0, THF

OF en ER

Me ) 3. Hy, PU/C EtOH

Chem Pharm Bull. v 40 (6) 1992, 1443-51. Chem Pharm Bull. v36 (10) 1988, 4121-4135. soc (7 1 XRX

H NSO, NN —_—

POA OH 2. TFA CHCl,

Me

H ALJ

HNSO, NA - Ses

R y ( } C 0

Cl

YO

OMe

Step 1. A solution of 1-(2-aminoethoxy)—2-benzyloxybenzene (Chem.

PFarm. Bull. 1988, 36, 4121-35; 1 mmol) in methanol (4 mL) is acidified with acetic acid to pH 6 .6 (pH meter) under a nitrogen atmosphere. “The ketone, 1-(4- methyoxy-3-amino sulfonyl)phenyl-2-oxopropane (Chem. Pharm. Bull. 1992, 40, 1443-51; 1 mmol) is added neat followed by sodium cyanoboroh ydride (1.1 mmol).

The course of the reaction is followed by thin layer chromatogra-phy. After reaction occurs, thee reaction solution is quenched in water and tkae pH of the aqueous mixture is- adjusted to 9-10 with aqueous Na,CO5. The mixture is extracted with ethesr, the organic extracts are washed with half-saaturated saline, : dried (Na,SO,), filtered and concentrated under reduced pressur « to give the crude product. The desimed compound is obtained by purification of thx.e crude product by use of HPLC.

Step 2. A solution of BOC-anhydride (5 mmol) and triethylamine (0.1 mL) in CH,Cl, (5 mL) is stirred under an inert atmosphere. To this is added a solution the product of the preceding reaction (2 mmol) in CH,Cl, (2 mL) and the resulting solution is stirred. The progress of the reaction is followed by TLC and when complete, the reaction is quenched by the addition of aqueous Naa,CO;. The mixture is extracte-d with CH,Cil,, the organic extracts are washed wiih haif- saturated saline, dried (Na,SO,), filtered and concentrated unde x reduced pressure to give the crude pproduct. The desired N-BOC-product is obtairied by purification of the crude product by use of HPLC.

Step 3. A s<olution of the compound from the preceding reaction in ethyl alcohol (10 mL) iss hydrogenated at atmospheric pressure in the presence of 10% palladium-on-carb on (100 mg) until tlc evidence shows that reacztion is complete.

The mixture is filcered through Celite and the filter pad is washed thoroughly with ethanol. The commbined filtrates are concentrated under reduced. pressure (0 give the crude product = The desired compound is obtained by purificcation of the crude product with the tase of HPLC.

Step 4. Diethyl azodicarboxylate (2 mmol) is added dropwise via a syringe to a stirred solution of triphenylphosphine (2 mmol) in THF (5 mL) at room temperature. To this is added a solution of the product from the preceding reaction (2 mmol) and 2,2’-(1,2-phenylenedioxy)diethanol (I mmol) in THF (3 mL). The resulting solution is stirred at RT and the progress of the reaction is followed by tlc. After reaction occ urs, solvent is removed by evaporation under reduced pressure and the residue is purified by HPLC, giving the desired compound.

Step 5. A solution of the product from the preceding reaction and trifluoroacetic acid (3 mL) in CH,Cl, (5 mL) is stirred at room temperature. The progress of the reaction is followed by tlc. After reaction occurs, more CH,Cl, is added and the solution is washed with aqueous Na,COj3 and with H,0. The organic layer is dried (INa,SO,), filtered and concentrated under reduced pressure to give the crude produ ct. The desired Formula I compound (where R is 1,2- phenylenedioxyethylene) is obtained by purification of the crude product with the use of HPLC.

-©6-

EXAMPLE 3: Preparation of a Formula I compound wherein p is 2, q is 1, and the ligand, L, is tamsulosin linked via the su 1fonamide group to the linker, X.

Example 3: Tamsulosin Analogs 1. (8OC)0, ~~ 00 0)

HN" a 2.1,5-dibromopentane, DMF ~ CH, 3. TFA (0) lo)

SH H 00 «a Qf H ~ XY)

LCL ' JO ~o

EH o H 0 CH

R=(CHy)s

Backgound Chemistry: 1. CISOH AP ~ 2. NH, ~o 0

Chem Pharm Bull. v40 (6) 1992, 1443-1451.

Reductive alkylation can be accomplished with chiral borane ligands: ~0 0. 8° HN )g oP pe )

HN oY ~NG LONE —_— z (o}

No NF HB(LY) Sg Ch

See Hett, Robert; Fang, Qun Kevin; Gao, Yun; Hong, Yaping: Butler, Ha! T.; et al Tetrahedron Lett. 38; 7, 1997; 1125-1128, for the use of chiral oxazaborolidirees in the enatioselective reductive alkylation of 2-phenyl ketones

Step 1. A solution of BOC-anhydride (5 mmol) and triethylamine (0.1 mL) in CH,Cl, (5 mL) is stirred under an ine rt atmosphere. To this is added a solution of tamsulosin (2 mmol) in CH,Cl, (2 mI) and the resulting solution is stirred. The reaction is followed by TLC and when complete, is quenched by the addition of aqueous Na,CO5. The mixture is extracted with CH,Cl,, the organic extracts are washed with half-saturated saline, dried (Na,S0,), filtered and concentrated under reduced pressure to give the crude product. The desired N-BOC-tamsulosin is obtained by purification of the crude product by use of HPLC.

Step 2. A mixture of NaH (2.1 mmol) and DMF (1 mL) is prepared umader an inert atmosphere in a flask equipped with a stirring bar and a drying tube. To this is first added a solution of N-BOC-tamsulosin (2 mmol) in dry DMF (3 mal) : followed by the linker molecule, 1,5-dibromopentane (1 mmol), in dry DMF «1 mL). The resulting mixture is stirred and the course of the reaction is followe=d by thin layer chromatography. After reaction occurs, the reaction is quenched with cold dilute aq. Nay COj3 and extracted with methylene chloride. The organic layer is dried (Na,S0,), filtered and concentrated under reduced pressure to give the crude product. The desired compound is obtained by purification of the crude product by use of HPLC.

Step 3. A solution of the product (2 mmol) of the preceding reaction and trifluoroacetic acid (3 mL) in CH,CI, (5 mL) is stirred at room temperature. “The progress of the reaction is followed by tlc. After reaction occurs, more CH,C lis added and the solution is washed with aqueous Na,COj; and with H,O. The organic layer is dried (Na,SO,), filtered and concentrated under reduced pressure to give the crude product. The desired Formula I compound [where R is -(CHRL,)s- ] is obtained by purification of the crude product with the use of HPLC.

EXAMPLE 4: Preparation of a Formula I compound wherein p is 2, q is 1, and the ligand, L, is (R)-KMD-3213 linked via the amide group to the linker, X.

Example 42 KMD-3231 Analogs 0. NH \-0oH °°" \__oH

CHa Ny 1.KOH, MeOH CHa N

Cr 2.(BOC)0 CU : 07k, oer

KMD-3213

F3C_O

H

~ i J pel 1. DCC, HOB, DIEA H HN °

H2N-g-NH2 i gh R = -(CHy)r-

Ny \-oH —_— hs 2. TFA On" ( [ H 0" CF,

Step 1. A solution of KMD-3213 (E.P. 0600 675 BI; 4 mmol) in methanol (4 mL) and aqueous 1M KOH (8 mL) is stirred under an inert atmosphere and is warmed until reaction occurs. The progress of the reaction is foliowed by tic.

When reaction is complete, the pH of the solution is adjusted to between 1 and 2 by the addition of 1 N HCI. The solution is then lyophilized and the crude reaction product is dried and used directly in the next step described below.

Step 2. A solution of BOC-anhydride (3 mmol) and triethylamine (0.15 mL) in CH,Cl, (5 mL) is stirred under an inert atmosphere. To this is added a solution the product of the preceding reaction (3 mmol) in CH,Cl, (2 mL) and the resulting solution is stirred. The progress of the reaction is followed by TLC and when complete, the reaction is quenched by the addition of dilute aqueous HCI. The mixture is extracted with CH,Cl,, the organic extracts are washed with half- saturated saline, dried (Na,SO,), filtered and concentrated under reduced pressure

) | WO 99/64042 PCT/UF S99/12728 to give the crude product. The desired N-BOC-product is obtained by pu tification of the crude product by use of HPLC.

Step 3. A solution of the product (2 mmol) from the preceding reaction, 1,7- diaminohe=ptane (1 mmol), and 1-hydroxybenzotriazole (2.5 mmol) in dry DMF (5 mL) is cooled in an ice-water bath and stirred under an inert atmosphere. To the stirred solution is added dicyclohexylcarbodiimide (2.1 mmol). The coursse of the reaction is followed by tlc. The cooling bath is removed and after reactiomn occurs, the reaction mixture is partitioned between methylene chloride and saturat-ed aqueous NaHCO,. The organic layer is washed with water and brine, drieed and concentrated under reduced pressure. The desired product is obtained by purificatiosn of the crude product by use of HPLC.

Step 4. A solution of the product from the preceding reaction (1 mrmol) and trifluoroacetic acid (3 mL) in CH,Cl, (5 mL) is stirred at room temperatuxe. The progress o-f the reaction is followed by tic. After reaction occurs, more CH,Cl, is added and the solution is washed with aqueous Na,CO3 and with H,O. The organic lawyer is dried (Na,SO,), filtered and concentrated under reduced pwressure to give the= crude product. The desired Formula I compound (where R is EB 7- heptanediy~1) is obtained by purification of the crude product with the use of HPLC.

The Formula I compound prepared by this method is obtained as a racemate when racemmic KMD-3213 is used in Step 1.

EXAMPLE 5: Preparation of a Formula I compound wherein p is 2, qis 1, and the ligand, L, is (R)-5-[2-[N-BOC-2-[2-(2,2,2- trifluoroethoxy)phenoxy]ethylamino)propyl]indoline-7—carboxamide [N-des-(3- hydroxypropyl)-KMD-3213] linked via the amino grou p of the dihydroindole to the linker, X.

Example 5: KMD-3231 Analogs

Ox -NHz 1. DMF, 50 C on Ar N I dibreomo-3-pentanol-O-TEDMS

Op —_—

CC

See Eur Pat Applic. EP 600675, EP 93-309450_931126

Og NH, HoN.__O

On ho

CL Pee

R= -CH,CH,CH(OH )CH,CH,-

Preparation of 1,5-dibrome-3-pentanol-O-TBDMS. tert-Butyldimethylsilyl chloride (0.1 mol) is addled to a solution of 1,5- dibromo-3-pentanol (0.05 mol) and imidazole (0.05 mol) in dry pyridine (10 mL) and the resulting solution is stirred at RT. The progress of the reaction is followed by tle. When reaction is complete, water (25 mL) is a«dded to the solution which is then concentrated by evaporation under reduced pressumre (>25 mm Hg, 30°C).

The residue is dissolved in EtOAc and the solution is extracted with saturated aq.

CuSQy, to remove residual pyridine. The EtOAc solution is washed with water, dried (Na,S0,), filtered and concentrated under reduced pressure to give the crude product. The pure product is obtained by purification of the crude product by flash chromatography over silica gel.

Compound Preparation.

Step 1. A solution of (R)-5-[2-[N-BOC-2-[2-(2, 2,2- trifluoroethoxy)phenoxy]ethylamino]propyllindoline-7 -carboxamide (EP 0600675, the (R), (S), and racemic forms of this compound may be found as the first three entries in the table on page 24 of this EP Specification. ; 2 mmol) and 1,5-dibromo- 3-pentanol-O-TBDMS (1 mmol), and diisopropylethylamine (0.2 mL) in DMF (3 mL) is stirred and warmed under an inert atmosphere. The progress of the reaction is followed by tic and when reaction is complete, the s olution is poured into aqueous 5% NaHCO; and the aqueous mixture is extracted with methylene chloride. The organic extract solution is dried (Na,SO), filtered and concentrated under reduced pressure to give the crude product. The desired compound is obtained by purification of the crude product by use of” HPLC.

Step 2. A solution of the compound prepared by~ the preceding reaction and trifluoroacetic acid (3 mL) in CH,Cl, (5 mL) is stirred. at room temperature. The progress of the reaction is followed by tlc. After react ion occurs, more CH, Cl, is added and the solution is washed with aqueous Na,CO— and with H,O. The organic layer is dried (Na,SO,), filtered and concentra ted under reduced pressure to give the crude product. The desired Formula I comgpound [where R is -CH,CH,CH(OH)CH,CH,-] is obtained by purification of the crude product with the use of HPLC.

The Formula I compound prepared by this method is of the (S) configuration in both ligands when (S)-KMD-3213 is u sed in Step 1. Likewise, the

Formula 1 compound of (R) configuration in both ligarads is obtained when (R)-

KMD-3213 is used in Step 1. The Formula 1 compound is obtained as a racemate when racemic KMD-3213 is used in Step 1.

By replacing the 1,5-dibromo-3-pentanol-O-TBDMS of the preceding example with other linker molecules, other compounds of Formula I are prepared.

EXAMPLE 6: Preparation of a Formula I compound wherein p is 2, q is 1, and the ligand, L, as (R)-KMD-3213 linked via the hydroxyl group to the Linker, X.

Exampmle 6: KMD-3213 Analogs

Oy NHe 1. TMSCI/Py/Imid

CHa N 3. EtgN-(HF)y

Cry 4. NaH/DMF 0 CF, KMD-3213 5 Ti diogobutane

See Eusr Pat Applic. EP 600675, EP 93-309450, -931126

Og NH, HNO rm \0_ _o/ chs N R N CHa

Cy NO

H R= 4. — 0" CF, (CHa) Ec oY

Step 1. Trimethylsilyl chloride (4 mmol) is added to a solution of (R)-KMD- 3213 (2 mmnol) and imidazole (2 mol) in dry pyridine (5 mL) and the resulting solution is stirred at RT. The progress of the reaction is followed by tlc. When reaction is ecomplete, water (5 mL) is added to the solution which is then concentrated by evaporation under reduced pressure (>25 mm Hg, 30°C). The residue is diissolved in EtOAc and the solution is extracted with saturated aq.

CuSO, to r emove residual pyridine. The EtOAc solution is washed with water, dried (Na, SO), filtered and concentrated under reduced pressure €o give the crude product. T he pure product, (R)-KMD-3213-0-TMS, is obtained by purification of the crude product by flash chromatography over silica gel.

Step 2. A mixture of (R)-KMD-3213-O-TMS (2 mmol), di-Zer:- butylcarborate (2.5 mmol), dioxane (5 mL) and aq. 2 N NaOH (2 ml) is stirred at

RT for 24 kar. The dioxane is removed by evaporation under reduced pressure.

Water (50 mL) is added tos the aqueous mixture and the mixture is extracted with

CH5Cl, (4 x 25 mL). The combined organic layers are dried (Na,80,), filtered and concentrated under reciuced pressure to give the crude product. Pure (R)-N—

BOC-KMD-3213-O0-TMS is obtained by purification of the crude product with tthe use of flash chromatography over silica gel.

Step. 3 A solution of (R)-N-BOC-KMD-3213-0-TMS (2 mmol), prepareci by the preceding reaction, and Et;N-(HF); in MeCN (5 mL) is stirred at room temperature. After reactio n occurs as detected by tic, the solution is diluted with

EtOAc and then washed w3th water-brine. The organic layer is dried (Na,S0,), filtered and concentrated under reduced pressure to give the crude product. The- desired compound (R)-N-B-OC-KMD-3213 is obtained by purification of the crucle product with the use of HPLC.

Step 4. A mixture o f NaH (2.1 mmol) and dry DMF (1 mL) is prepared under an inert atmosphere dn a flask equipped with a stirring bar and a drying tule.

To this is added a solution of the compound (2 mmol) prepared by the preceding reaction in dry DMF (3 mL). Then a solution of 1,4-diiodobutane (1 mmol) in clry

DMF (2 mL) is added and the resulting mixture is stirred, warmed and monitore=d for reaction by tlc. After reaction occurs, the reaction solution is quenched water (25 mL) and brine (25 mL@. The mixture is extracted with CH,Cl, (4 x 20 mL) and the combined organic extracts are back-washed with water (3x). The organi c layer is dried (Na,SOy,), filltered and concentrated under reduced pressure to gives the crude product. The desired compound is obtained by purification of the crue product with the use of HP-LC.

Step 5. A solution of the product from the preceding reaction and trifluoroacetic acid (3 mL) in CH,Cl, (5 mL) is stirred at room temperature. The progress of the reaction is #ollowed by tlc. After reaction occurs, more CH,Cl, is added and the solution is washed with aqueous Na,CO; and with H,0. The organic layer is dried (Na,SQ,), filtered and concentrated under reduced pressure to give the crude product. The desirec Formula I compound (where R is -(CH,)4- ] is obtained by purification of the cruade product with the use of HPLC.

S The Formula I compound prepared by this method is obtained as a racemate when racemic KMD-3213 is used in Sgtep 1. The Formula 1 compound with ligands of (S) configuration is obtained when (S)-KMD-3213 is used in Step 1.

EXAMPLE 7: Preparation of a Forme 1a I compound wherein p is 2, q is 1, and the ligand, L, is the N-(3-Hydroxy)propyl—7-aminocarbonyl-5-(2- amino)propyldihydroindole moiety of BKMD-3213 linked via the amino group to the linker, X.

Example 7: KMD-3213 Analogs

CN 1.5 N NaOH On-NHe

JUS 2. 30% HyCO,/DMSO Sais

Ebb ati CH, : soon SCL) 4. TMSClii mid/Py 5. Hyp, PUC

CAS 160968-97-8

See Eur Pat Applic. EP 600675, EP 93—309450_931126

HNO Ox NH, 1. 1,2-bis(2-bromoethoxy)oenzens Ho—/ [\—oH . SE A— N CH, CH N :

DMF LR. 2. HOAGH,0 NTN -CH,CH, H ne -CH,CH,0 Eg

Step 1. A solution of l-acetyl-5 —(2-azidopropyl)indoline-7-carbonitrile (EP 0 600 675 BI; 2 mmol) in aqueous 5 N NaOH (2 mL) and ethanol (4 mL) is stirred at room temperature. The course of the meaction is followed by tlc and when complete, the reaction solution is made= alkaline by the addition of cold 1 N NaOH.

The mixture is extracted with ether, th e ether extracts are washed with water and with brine, dried (Na,SOy), filtered and concentrated under reduced pressure to give the crude product. The desired 5-(2-azidopreopyl)indoline-7-carbonitrile is obtained by purification of the crude product witha the use of HPLC.

Step 2. To a solution of 5-(2-azidopropyl)imdoline-7-carbonitrile (2 mmol) in

DMSO (3 mL) is added 30% hydrogen peroxide (0.5 mL). The resulting mixture is stirred at room temperature for 20 minutes and then is transferred into a solution of aqueous 5 N NaOH (0.5 mL). This mixture is stirred at room temperature and the progress of the reaction is followed by tic. When complete, the mixture is neutralized by the addition of acetic acid, water is added, and the mixwre is extracted with ethyl acetate. The organic layer is washed with dilute aqueous sodium carbonate, with water and then is dried, fi Itered and concentrated under reduced pressure. The desired 5-(2-azidopropyl)imdoline-7-carboxamide is obtained by purification of the crude product with the use of HPLC.

Step 3. A solution of 5-(2-azidopropyl)indoline-7-carboxamide (2 mmol) together with 3-bromopropan-1-ol (2 mmol) in dioxane is heated and stirred with potassium carbonate. The progress of the reaction is followed by tlc and when comglete, the sclvent is removed by evaporation tancer reduced pressure. The residue is partitioned between dilute aq. sodium bicarbonate and ethyl acetate. The organic extract layer is washed with water and with brine, is dried (Na,SO,), filtered and concentrated under reduced pressure t o give the crude product. The desired 1-(3-hydroxypropyl)-5-(2-azidopropyl)indeline-7-carboxamide is obtained by purification of the crude product with the use af HPLC.

Step 4. Trimethylsilyl chloride (4 mmol) is added to a solution of 1-(3- hydroxypropyl)-5-(2-azidopropyl)indoline-7-carboexamide (2 mmol) and imidazole © 25 (2 mo)) in dry pyridine (5 mL) and the resulting solution is stirred at RT. The progress of the reaction is followed by tic. When reaction is complete, water (5 mL) is added tO the solution which is then concentrated by evaporation under reduced pressume (>25 mm Hg, 30°C). The residue is dissolved in FEtOAc and the solution is extracted with saturated aq. CuSO, to remove residual pyridine. The

EtOAc solutiore is washed with water, dried (Na,SO,), filtered and cowncentrated under reduced pressure to give the crude product. The pure 1-(3-hydxoxypropyl)- 5-(2-azidopropy/l)indoline-7-carboxamide-O-TMS is obtained by purification of the crude product bay flash chromatography over silica gel.

Step 5. A\ solution of 1-(3-hydroxypropyl)-5-(2-azidopropyl)incloline-7- carboxamide-O—-TMS in ethyl acetate (10 mL) is hydrogenated at atmcespheric pressure in the presence of 10% palladium-on-carbon (100 mg) until tc evidence shows that reaction is complete. The mixture is filtered through Celite and the filter pad is washed thoroughly with ethyl acetate. The combined filtrates are concentrated urmder reduced pressure to give the crude product. The esired 1-(3- hydroxypropyl)--5-(2-aminopropyl)indoline-7-carboxamide-O-TMS is ©btained by purification of the crude product with the use of HPLC.

Step 6. AA solution 1-(3-hydroxypropyl)-3-(2-aminopropyl)indoine-7- carboxamide-®—TMS (2 mmol) and 1,2-bis(2-bromoethoxy)benzene (Aldrich; 1 mmol), and diisopropylethylamine (0.2 mL) in DMF (3 mL) is stirred and warmed under an inert atmosphere. The progress of the reaction is followed bey tlc and when reaction i s complete, the solution is poured into aqueous 5% NaHCO; and the aqueous mixture is extracted with methylene chloride. The organic extract solution is driecd (Na,SO,), filtered and concentrated under reduced pxressure to give the crude gproduct. The desired compound is obtained by purification of the crude product by use of HPLC.

Step 7. A solution of the product of the preceding reaction in FHOAc-H,0 (4:1) (5 mL) is stirred under an inert atmosphere at room temperature=. The weaction is followed by tic and when complete, is diluted with EtOAc and washed several times with water and dilute aq. Na,COj;. The organic layer is dried (Na,S0y,), filtered and concentrated under reduced pressumre to give the crude product. The desired Formula I compound is obtained by purification of the crude product by use of HPLC.

EXAMPLE 8: Preparation of a Formula I compound wherein p is 2, q is 1, and the lx gand, L, is the N-(3-Hydroxy)propyl-7-aminocarbonyl-5—(2- ammino)propyldihydroindole moiety of KMD-3213 linked via the amino group to a bas-catechol linker, X. © Example 8: KMD-3213 Analogs 1. DEAD

TPP Br

OC Cl 22, 000

OH 2. HyPAC oF Lor on 0 Ro

Og

CN

1. OUNH, Ho \_oTms CH,

CH, N NH -0

J R= {CH,);- J

DIPEA bi ON NC oH 2. HOAG-H,0 o oY ~T-N

H

Pr eparation of 1,5-Bis[o-(2-bromoethoxy)phenoxy]pentare.

Steep A. Diethyl azodicarboxylate (2 mmol) is added ropwise via a syringe to = stirred solution of triphenylphosphine (2 mmol) in THF (5 mL) a1 room teriperature. To this is added a solution of o-benzyloxyphemol (2 mmol) and 2- brosmoethanol (2 mmol) in THF (1 mL). The resulting solu®ion is stirred at RT and the progress of the reaction is followed by tlc. After reactio=n occurs, solvent is rercioved by evaporation under reduced pressure and the resi_due is purified by

HPLC, giving the desired 1-benzyloxy-2-(2-bromoethoxy)be=nzene.

Step B. A solution of 1-benzyloxy-2-(2-bromoethoxy)bermzene in ethyl acetates (10 mL) is hydrogenated at atmospheric pressure in the gpresence of 10% pallad ium-on-carbon (100 mg) until tlc evidence shows that reaction is complete.

The mixture is filtered through Celite and the filter pad is washed thoroughly with ethyl ;acetate. The combined filtrates are concentrated under recluced pressure to give the crude product. The desired o-(2-bromo)ethoxyphenol 3s obtained by purification of the crude product with the use of HPLC.

Step C. Diethyl azodicarboxylate (2 mmol) is added drop»wise via a syringe to a s=irred solution of triphenylphosphine (2 mmol) in THF (5 mL) at room tempe=rature. To this is added a solution of 0-(2-bromoethoxy)phenol (2 mmol) and pentam-1,5-diol (1 mmol) in THF (1 mL). The resulting solution is stirred at RT and the progress of the reaction is followed by tlc. After reaction occurs, solvent is remowed by evaporation under reduced pressure and the residue is purified by

HPLC, giving the desired 1,5-bis{o-(2-bromoethoxy)phenoxy]pentane.

Comppound Preparation

Step 1. A solution 1-(3-hydroxypropyl)-5-(2-aminoprop¥yl)indoline-7- carbosxamide-O-TMS (2 mmol), 1,5-bis[o-(2-bromoethoxy)phemoxy]pentane (1 mmo 31), and diisopropylethylamine (0.2 mL) in DMF (3 mL) is stirred and warmed undex an inert atmosphere. The progress of the reaction is folk owed by tlc and wherm reaction is complete, the solution is poured into aqueous 5% NaHCO; and the aqueous mixture is extracted with methylene chloride. The organic extract solutdon is dried (NaySO,), filtered and concentrated under recluced pressure to give the crude product. The desired compound is obtained by purification of the crude product by use of HPLC.

Step 2. A solution of the product of the preceding react ion in HOAc-H,0 (4:1 (5 mL) is stirred under an inert atmosphere at room temgoerature. The

’ © WO 9964042 PCT/US99/12728 reaction is followed by tic and when complete, is diluted with EtOAc and washed several times with water and dilute ag. Na,CO;. The organic layer is dried (Na,S0y), filtered and concentrated under reduced press=ure to give the crude product. The desired Formula I compound is obtained b-y purification of the crude

Jproduct by use of HPLC.

EXAMPLE 9: Preparation of a Formula I compound whesrein p is 2, q is 1, and the

Rigand, L, is 4-amino-2-(N-piperazinyl)-6,7-dimethoxy-2—quinazoline linked through the piperazine amino group via an amide to the lanker, X.

Example 9: Cardura Analogs (NH

For P, : Sekiya, Tetsuo; Hir = . Hidetoshi; Hata,

MeO No Shunsoke: Mizogam. Susamor Hapazake, Mitsuo: Vameds, _N Shun-ichi J.Med.Chem.: 26: 3: 19833: 411-416.

MeO

NH,

XK

0 0

NT RTON

Ho ron MeO Ny i$ C3 _N OMe yy hig > MeO ~N NS OMe

NH, NH,

MeO, OMe

R= Dy R preferably contains alkyl, alkyl-ether linkages, hy—droxy! grps. hydrogen bond acceptors, aryoxyl. heteroaryl groups

A solution of 4-amino-2-(N-piperazinyl)-6,7-dime®hoxy-2-quinazoline (2 mmol), 2,3-dimethoxybenzene-1,4-dicarboxylic acid (1 mamol), and 4- & imethylaminopyridine (10 mg) in CH,Cl, (5 mL) is prepared under argon in a flask equipped with magnetic stirrer and a drying tube. To this solution is added d icyclohexylcarbodiimide (solid, 2.2 mmol). The progress of the reaction is followed by tlc and after reaction occurs, the reaction soltation is quenched in water, aqueous sodium bicarbonate is added and the aqueous mixture is extracted with methylene chloride. The organic layer is washed wish aqueous Na,CO4 and with Hy0, dried (Na,SO,), filtered and concentrated under reduced pressure to give the crude product. The desired Formula I compound is obtained by purification of the crude product with the use of HPLC.

EXAMPLE 10: Preparation of a Formula I compound wherein p is 2, qis1, and the ligand, L, is 4-amino-2-(N-piperazinyl)-6,7-dimethoxy-2-quuinazoline linked throug h the piperazine amino group via a carbamate to the linker, X.

Example 10: Cardura Analogs ne. 1 ] ] ]

HOR" NoH Tw PRP Ap hg PNP oerdT a hk [NTTOTTRTTOT NTT

Meo I MeO pt LN eS <$ oe

DMAP, HOBY, DMF, 1t 00 NH, R = -CH,OCH,- NH, oe

R preferably Contains alk. alkyl-ether linkages, hydroxyl grps, hydrogen bond acceptars,

A solution of diethylene glycol) (2 mmol) and pyridine (2.5 mmol) in dry

THF («4 mL) is cooled (ice-water bath) and stirred under an inext atmosphere. To this a solution of 4-nitrophenylchloroformate (2.1 mmoi) in dry THF (1 mL) is added dropwise. After the addition is completed, the ice-water cooling bath is removed and the solution is stirred and allowed to reach ambient temperature. The reaction is followed by tlc and after reaction is complete, THF is removed under reduced pressure. The residue is protected from atmospheric moisture and is dissolved in dry DMF (2 mL). To this solution are added 4-dir nethylaminopyridine (1 mg», 1-hydroxybenzotriazole (3 mmol), and a solution of 4-amino-2-(N- piperazinyl)-6,7-dimethoxy-2-quinazoline (4 mmol) in dry DM¥ (2 mL). The resulting solution is stirred at room temperature and the progress of the reaction is follow ed by tic. When complete, the reaction solution is quenched by addition of cold aq. Na,CO4 and brine. The mixture is extracted with methylene chloride, the organi c layers are combined, washed with water and with brines, are dried (Na,SQy,), filtered and concentrated under reduced pressure to give the crude product. The desired ®Formula I compound (where R is -CH,CH,0CH, CH,-) is obtained by purification of the crude product by use of HPLC.

EXAMPLE 11: Preparation of a Formula I compound wherein ligand, L 1, the phenethylamine moiety~ of Tamsulosin is linked via the amino group to ligand, L,, the phenylbutoxyhexyl moiety of Salmeterol.

Example 11: Salmetero | analogs

AP Cy AP NHEn

HN HN"

OY NaCNBH3, MeOH SAN ohhh. 00 2. Hp, PdIC, Et OH

Step 1. A solutio=n of benzylamine (2 mmol) in methanol (4 mL) is acidified with acetic acid to pH 6 .6 (pH meter) under a nitrogen atmosphere. The ketone, 1-(4-methyoxy-3-aminossulfonyl)phenyl-2-oxopropane (Chem. Pharm. Bull. 1992, 40, 1443-51; 2 mmol) is added neat followed by sodium cyanoborohydrid e (2.1 : 10 mmol). The course of tkae reaction is followed by thin layer chromatograp hy. After reaction occurs, the reaction solution is quenched in water and the pH of =he aqueous mixture is adjusted to 9-10 with aqueous Na,CO5;. The mixture 3s extracted with ether, thes organic extracts are washed with half-saturated saline, dried (Na,SO,), filtered. and concentrated under reduced pressure to give the crude product. The desired compound is obtained by purification of the crude p roduct by use of HPLC.

Step 2. A solutiom of the product of the preceding reaction and n-(4&- phenyl)butyl-n-(6-brome)hexyl ether (or 1-bromo-11-phenyl-7-oxaundecare) 2 mmol), and diisopropylesthylamine (0.2 mL) in DMF (3 mL) is stirred andl warmed under an inert atmosphemre. The progress of the reaction is followed by tics and

WG 99/64042 PCT/US99/12728 -1 12- when reaction is complete, the solution iss poured into aqueous 5% NaHCO; and the aqueous mixture is extracted with methylene chloride. The organic extract solution is dried (Na,SQ,), filtered and concentrated under reduced pressure to give the crude product. The desired comapound is obtained by purification of the crude product by use of HPLC.

Step 3. A solution of the compournd from the preceding reaction in ethanol (5 mL) is stirred under hydrogen at atmo spheric pressure in the presence of 10% palladium-on-carbon (100 mg) until tic ewidence shows that reaction is complete.

The mixture is filtered through Celite ancl the filter pad is washed thoroughly with ethyl acetate. The combined filtrates are concentrated under reduced pressure to give the crude product. The desired Foremula | compound is obtained by purification of the crude product with thes use of HPLC.

EXAMPLE 12: Preparation of a Formulaa I compound whercin ligand, L, the dihydroindole moiety of KMD-3213 is limked via the amino group to ligand, L,, the phenylbutoxyhexyl moiety of Salmetesrol.

Example 12: Salmeterol analogs [o] on on " on Sp

CH, N = CH3

HN NaCNB- H3, MeOH BrHN

Ohhh: 0 ( 2. Hj, PIC, EtOH Coy : 3. HOAc

For Prep see: Hett, Robert; Stare, Ragnar; Helcquist, Paul Tetrahedron Lett.: 35; 50; 1994; 9375-9378.

Step 1. A solution of 1-(3-hydroxsypropy!)-5-(2-aminopropyl)indoline-7- carboxamide-O-TMS (2 mmol) in metharol (4 mL) is acidified with acetic acid to pH 6.6 (pH meter) under a nitrogen atmosphere. Benzaldehyde (2.2 mmol) is added neat followed by sodium cyanoborohydride (2.1 mmol). The cou rse of the reaction is followed by thin layer chromatography. After reaction occas, the reaction so-fution is quenched in water and the pH of the aqueous mixture is adjusted to 9-10 with aqueous Na,CO;. The mixture is extracted with ether, the organic ex®&racts are washed with half-saturated saline, dried (Na,SO,), filtered and concentrated under reduced pressure to give the crude product. The de sired compound is obtained by purification of the crude product by use of HPLC.

Step 2. A solution of the product of the preceding reaction and re=-(4- phenyl)butzyl-n-(6-bromo)hexyl ether (or 1-bromo-11-phenyl-7-oxaunde cane) (2 mmol), and diisopropylethylamine (0.2 mL) in DMF (3 mL) is stirred &and warmed under an irxert atmosphere. The progress of the reaction is followed by tlc and when react ion is complete, the solution is poured into aqueous 5% NaHICO; and the aqueous mixture is extracted with methylene chloride. The organic extract solution is dried (Na,SQO,), filtered and concentrated under reduced pressure to give the crude product. The desired compound is obtained by purificat ion of the crude product by use of HPLC.

Step 3. A solution of the compound from the preceding reaction in ethanol (5 mL) is stirred under hydrogen at atmospheric pressure in the presence of 10% palladium-on-carbon (100 mg) until tlc evidence shows that reaction is complete.

The mixture is filtered through Celite and the filter pad is washed thoroughly with ethyl acetate. The combined filtrates are concentrated under reduced p ressure to give the crude product. The desired compound is obtained by purification of the crude prod uct with the use of HPLC.

Step 4. A solution of the product of the preceding reaction in H&@OAc¢-H,0O (4:1) (5 ml) is stirred under an inert atmosphere at room temperature. The reaction is followed by tlc and when complete, is diluted with EtOAc and washed several times with water and dilute aq. Na,CO5. The organic layer is dried (Na,S0Qy), filtered and concentrated under reduced pressure to give the crude product. The desired Formula I compound is obtained by purification of the crude product by use of HPLC.

Examples 13 and 14: Mixed Dimers backgound Chemistry 0° Q 0

Ox -NH2 HANS cd Ox NH; \—otMs ~ 0 o oH

CH, N 0 » - fed Hy N

HN NaCNBH3, EtOH HaN-s 2 N x 2. HOAG-H,0 oo H 1. rn A Example 14

Ox -NH2 2 ES CH 2 \_orws N OY | Aon

HaN NaCNBH3, EtOH HaN-g N 2. HOAc-H;0 oo H also hormodéeners (see Ex. 15 & 16)

EXAMPLE 13: Preparation of a Formula I compound wherein ligand, L,, the dihydroindole moiety of KMD-3213 is linked via the amino group to ligand, L,, the phenethylamine moiety of Tamsulosin. (See Fig. above.)

Step 1. A solution of 1-(3-hydroxypropy!)-5-(2 —aminopropyl)indoline-7- carboxamide-O-TMS (2 mmol) in methanol (4 mL) is acidified with acetic acid to pH 6.6 (pH meter) under a nitrogen atmosphere. 1-(4-methyoxy-3- aminosulfonyl)phenyl-2-oxopropane (2 mmol) is added} neat followed by sodium cyanoborohydride (2.1 mmol). The course of the reaction is followed by thin layer

© WO 99/6404 PCT/US99/12728 chromatography. After reaction occuars, the reaction solution is quenched in water and the pH of the aqueous mixture is adjusted to 9-10 with aqueous Na,CO,. The mixture is extracted with ether, the omrganic extracts are washed with half-saturated saline, dried (Na,S0,), filtered and c oncentrated under reduced pressure to give the crude product. The desired compound is obtained by purification of the crude product by use of HPLC. :

Step 2. A solution of the product of the preceding reaction in HOAc-H,0 (4:1) (5 mL) is stirred under an inert saatmosphere at room temperature. The reaction is followed by tlc and when complete, is diluted with EtOAc and washed several times with water and dilute aq . Na,CO5. The organic layer is dried (Na,80,), filtered and concentrated u nder reduced pressure to give the crude product. The desired Formula I compound is obtained by purification of the crude product by use of HPLC.

EXAMPLE 14: Preparation of a Formula I compound wherein ligand, L |, the dihydroindole moiety of KMD-3213 is linked via the amino group to ligand, L,, the desmethyl-phenethylamine moiety of Tamsulosin. (See Fig. above.)

Step 1. A solution of 1-(3-hydr oxypropyl)-5-(2-aminopropyl)indoline-7- carboxamide-O-TMS (2 mmol) in met hanol (4 mL) is acidified with acetic acid to pH 6.6 (pH meter) under a nitrogen atmosphere. 1-(4-methyoxy-3- aminosulfonyl)phenylacetaldehyde (2 mmmol) is added neat followed by sodium cyanoborohydride (2.1 mmol). The ceourse of the reaction is followed by thin layer chromatography. After reaction occurs, the reaction solution is quenched in water and the pH of the aqueous mixture is adjusted to 9-10 with aqueous Na,CO5. The mixture is extracted with ether, the or ganic extracts are washed with half-saturated saline, dried (Na,SQy), filtered and concentrated under reduced pressure to give the crude product. The desired compound is obtained by purification of the crude product by use of HPLC.

Step 2. A solution «of the product of the preceding reaction in HOAc-H,0 (4:1) (5 mL) is stirred uncer an inert atmosphere at room temperature. The reaction is followed by tlc and when complete, is diluted with EtOAc and washed several times with water a nd dilute ag. Na,CO;. The organic layer is dried (Na,SO,p), filtered and corxcentrated under reduced pressure to give the crude product. The desired Foramula I compound is obtained by purification of the crude product by use of HPLC.

EXAMPLE 15: Preparation of a Formula I compound wherein p is 2, q is 1, and the ligand, L, is the compound prepared in Example 13 linked through the chain amino group to the linker, X.

Example 15 [o} NH» 1. 2,2-(1,2-phenylenedioxy)diethylbromide

HN N x 2. HOAG-H,0 00 H O._NH,

HN.

Ne &o ~~. Ox NH; -CHCSHR0 “OLE

HaN-52 N oo

Step 1. A solution ©f the product from Example 13, Step 1 (2 mmol) and 2,2'-(1,2-phenylenedioxy) diethylbromide (1 mmol), and diisopropylethylamine (0.2 mL) in DMF (3 mL) is sti rred and warmed under an inert atmosphere. The progress of the reaction is followed by tic and when reaction is complete, the solution is poured into aqraeous 5% NaHCO; and the aqueous mixture is extracted with methylene chloride. The organic extract solution is dried (Na,SQy), filtered and concentrated under reduced pressure to give the crude product. The desired compound is obtained by pur ification of the crude product by use of HPLC.

Step 2. A solution of the product of the preceding reaction in HOAc-H,0 (4:1) (5 mL) is stirred under an inert atmosphere at room temperature. The

S reaction is followed by tlc and when complete, is diluted with EtOAc and washed several times with water and dilute aq. Na,CO5. The organic layer is dried (Na,S0,), filtered and concemtrated under reduced pressure to give the crude product. The desired Formuka I compound is obtained by purification of the crude product by use of HPLC. : 10 EXAMPLE 16: Preparation of a Formula I compound wherein p is 2, q is 1, and the ligand, L, is the compoun d prepared in Example 14 linked through the chain amino group to the linker, X.

Example 16 o NH 2 1. di(2-bromoethyl)ether o \-OTMs DIPEA ~ N

HaN-g- N 2. HOAc-H,0 o “0 H

Ox NH; o I \_-oH

HN

Ss N oo ~~

R Oa NH,

R = -CH,CH;0CH,CHy o NOH - [Wl ons N

Halo N oo .

Step 1. A solution of the product from Example 14, Step 1 (2 mmol) and 2- bromoethy! ether (1 mmol), and diisopropylethylamine (0.2 mL) in DMF (3 mL) is stirred and warmed under an inert atmosphere. The progress of the reaction is followed by tic and when reaction is complete, the solution is poured into aqueous

~118- 5% NaHCO; and the aqueous mixture is extracted with methylene chloride. The organic extract solution is dried (Na,S0Q,), filtered and concentrated under reduced pressure to give the crude product. The desired compound is obtained by purification of the crude product bs use of HPLC.

Step 2. A solution of the product of the preceding reaction in HOAc-H,0 (4:1) (5 mL) is stirred under an inert atmosphere at room temperature. The reaction is followed by tlc and when complete, is diluted with EtOAc and washed several times with water and dilute aq. Na,CO;. The organic layer is dried (Na,SOy), filtered and concentrated under reduced pressure to give the crude product. The desired Formula I compound is obtained by purification of the crude product by use of HPLC.

EXAMPLE 17: Preparation of a Formula I compound wherein pis2,qis 1, and wherein one ligand L,, is the dihydroindole moiety of KMD-3213 linked via the chain amino group to the linker, X , and the second ligand, L,. is the phenethylamine moiety of TamsulOsin linked to X via the amino group.

Example 17: Mixed Dimers a, 0 1. NaB(CN)HyMeOH

FENN WY _2_ PhCH;0C(O)CI?DMAP/Py ~0 CHa 3. TPAPINMO ; Oa NH; 0.0 BOC gy

OL BnHN ~o CHj _

NaB(CN)HyMeOH 2. HOAC-H0 3. HyPd-C :

Os NH, oH

H2No NT SRTN do H H

R =-CH,CHp-

-119e-

Step 1. A solution of 4-amino-1-butaanol (2 mmol) in methanol (4 mL) is acidified with acetic acid to pH 6.6 (pH me ter) under a nitrogen atmosphere. 1-(4- methyoxy-3-aminosulfonyl)phenyl-2-oxopropane (2 mmol) is added neat followed by sodium cyanoborohydride (2.1 mmol). “The course of the reaction is followed by thin layer chromatography. After reaction occurs, the reaction solution is quenched in water and the pH of the aqueous mixture is adjusted to 9-10 with aqueous NayCO5. The mixture is extracted with ether, the organic extracts are washed with half-saturated saline, dried (N=m,S0,), filtered and concentrated under reduced pressure to give the crude product. The desired compound is obtained by purification of the crude product by use of HPLC.

Step 2. A solution of the product of tthe preceding reaction (2 mmol) in pyridine (5 mL) containing 4-dimethylaminoOpyridine (2-10 mg) is cooled in an ice : bath and benzyl chloroformate (0.5 mL) is a.dded. The cooling bath is removed and the reaction solution is stirred at room tesmperature. Progress of the reaction is . 15 followed by tic and when complete, the reaction is diluted with ethyl acetate, - washed with 5% aq. sodium bisulfate, dried (Na,;SQy), filtered and concentrated under reduced pressure to give the crude product. The desired compound is obtained by purification of the crude product by use of HPLC.

Step 3. A solution of the product of thhe preceding reaction (2 mmol) and 4- methylmorpholine N-oxide (NMO, 3 mmol) in MeCN (10 mL) and methylene chloride (5 mL) is treated with 4-A molecular sieves (50 mg) and stirred at ambient temperature for 10 min. Tetrapropylammomium perruthenate (TPAP, 0.05 mmol) is added and the reaction mixture is stirred a.t room temperature. The progress of the reaction is followed by tlc and when comraplete, the reaction is diluted with methylene chloride and the mixture is filtere d through silica gel. The filtrate is concentrated, giving the crude product.

Step 4. A solution of the product from Example 12, Step 1 (2 mmol) in methanol (4 mL) is acidified with acetic acid to pH 6.6 (pH meter) under a nitwrogen atmosphere. To this is added a solution of the aldehyde (2 mmol) prepared by the preceding step in me-thanol (1 mL) followed by sodium cyanoborohydride (3

S mmol). The course of the reaction is followed by thin layer chromatography. _After reaction occurs, the mreaction solution is quenched in water and the pH of the aqueous mixture is aedjusted to 9-10 with aqueous Na,CO;. The mixture is extracted with ether, the organic extracts are washed with half-saturated saline , dried (Na,SO,), filteered and concentrated under reduced pressure to give the crude product. The desire compound is obtained by purification of the crude prod ct by use of HPLC.

Step 5. A soluation of the product of the preceding reaction in HOAc-H»O (4:1) (5 mL) is stirre=d under an inert atmosphere at room temperature. The reaction is followed By tlc and when complete, is diluted with EtOAc and waskhed several times with water and dilute aq. Na,CO5. The organic layer is dried (Na,S0,), filtered amd concentrated under reduced pressure 0 give the crude product. The desired compound is obtained by purification of the crude product by use of HPLC.

Step 6. A solution of the compound from the preceding reaction in ethy acetate (10 mL) is hydrogenated at atmospheric pressure in the presence of 10 % palladium-on-carbon_ (100 mg) until tlc evidence shows that reaction is comple te.

The mixture is filtered through Celite and the filter pad is washed thoroughly “with ethyl acetate. The ceombined filtrates are concentrated under reduced pressure 10 give the crude product. The desired Formula I compound is obtained by purification of the crude product with the use of HPLC.

Examples 18 and 19: Mixed Dimers i I as

NH HOP Rx e \ I$ RX

Me NM > Y

JOC HOB, DCC, DWF Me ZN

Me NH,

NH,

Oy -NH, \-oTMs 2° NHBn 1) DMF Xx RMX + CH N HN BORA _—

BnH OR Cr 2. H2, PW/C, EtOH 3. (for Ex. 18) HOAC-H,0

Oy NH, \_oH

Example 18 0 CH, N

RN

Me Note H

JCC R =(CH,)q- or

Me . NH,

Example 19 0 £00

ASR ek

Me NN H oo

JOC R= (Crile: . Me

NH,

EXAMPLE 18: Preparation of a Formula I compound wherein p is 2, Gis 1, and wherein one ligand L,, is the dihydroindiole moiety of KMD-3213 linked via the chain amino group to the linker, X, and the second ligand, L,, is the (4-amino-6,7- dimethoxy-2-quinazolinyl)piperazine mo iety of Cardura linked to X via the piperazine amino group. (See Fig. above.)

Step 1. A mixture of 4-amino-2-(IN-piperaziny1)-6,7-dimethoxy-2- quinazoline (2 mmol), dicyclohexylcarbodiimide (2.2 mmol), and 4- dimethylaminopyridine (10 mg) in CH,Cl, (5 mL) is prepared under nitrogen in a flask equipped with magnetic stirrer and a drying tube. To the stirred mixture is added a solution of 6-bromohexanoic acid (1 mmol) in CH,Cl, (3 mL). The mixture is stirred at room temperature and the progress of the reaction is followed by tlc. After reaction o ccurs, the reaction solution is quenched in water, aqueous sodium bicarbonate is added and the aqueous mixture is extracted with methylene chloride. The organic 1 ayer is washed with aqueous Na,CO; and with H,O, dried (Na,S0y), filtered and concentrated under reduced pressure to give the crude product. The desired compound is obtained by purification of the crude product with the use of HPLC.

Step 2. A solution of the product of the preceding reaction (1 mmol) and the dihydroindole derivative prepared in Example 12, Step 1 (1 mmol), and diisopropylethylamine (0.2 mL) in DMF (3 mL) is stirred and warmed under an inert atmosphere. The progress of the reaction is followed by tlc and when reaction is complete, the solution is poured into aqueous 5% NaHCO; and the aqueous mixture is extracted with methylene chloride. The organic extract solution is dried (Na,SO,), filtered and concentrated under reduced pressure to give the crude product. The desired compound is obtained by purification of the crude product by use of HPLC.

Step 3. A solution of the compound of the preceding reaction in ethyl acetate (10 mL) is hydrogenated at atmospheric pressure in the presence of 10% palladium-on-carbon (100 mg) until tlc evidence shows that reaction is complete.

The mixture is filtered through Celite and the filter pad is washed thoroughly with ethyl acetate. The combined filtrates are concentrated under reduced pressure to give the crude product. The desired compound is obtained by purification of the crude product with the rise of HPLC.

Step 4. A solution of the product of the preceding reaction in HOAc-H,0 (4:1) (5 mL) is stirred wnder an inert atmosphere at room temperature. The reaction is followed by tlc and when complete, is diluted with EtOAc and washed

) © WO099/64042 PCT/US99/12728 several times with water and dilute aq. INa,CO;. The organic layer is dried (Na;S0,), filtered and concentrated under reduced pressure to give the crude product. The desired Formula I compowind, where R is —-(CH,),-, is obtained by purification of the crude product by use of HPLC.

EXAMPLE 19: Preparation of a Formu la I compound wherein p is 2, q is I, and wherein one ligand L, is the phenethylzamine moiety of Tamsulosin linked via the chain amino group to the linker, X, and the second ligand, L,, is the (4-amino-6,7- dimethoxy-2-quinazolinyl)piperazine moiety of Cardura linked to X via the piperazine amino group. (See Fig. abowe.)

Step 1. A solution of the product prepared by the method of Example 17,

Step 1 (1 mmol) and 1-(4-methyoxy-3-a-minosulfonyl)phenyl-2-benzylaminopropane (prepared as in Example 11, Step 1; 1 mmol), and diisopropylethylamine (0.2 mL) in DMF (3 mL) is stirred and warmed wander an inert atmosphere. The progress of the reaction is followed by tlc and whem reaction is complete, the solution is poured into aqueous 5% NaHCO, and the aque=ous mixture is extracted with methylene chloride. The organic extract solution i_s dried (Na,S0,), filtered and concentrated under reduced pressure to give the crud e product. The desired compound is obtained by purification of the crude preoduct by use of HPLC.

Step 2. A solution of the compound of the preceding reaction in ethyl acetate (10 mL) is hydrogenated at atmospheric pressure in the presence of 10% palladium-on-carbon (100 mg) until tic evidence shows that reaction is complete.

The mixture is filtered through Celite a nd the filter pad is washed thoroughly with ethyl acetate. The combined filtrates ane concentrated under reduced pressure to give the crude product. The desired cosmpound is obtained by purification of the crude product with the use of HPLC,

Step 3. A solution of the product of the preceding reaction in HOAc-H,0 (4:1) (5 mL) is stirred under an inert atmosphere at room temperature. The reaction is followed by” tlc and when complete, is diluted with EtOAc and washed several times with water and dilute aq. Na,CO3. The organic layer is dried (Na,SO,), filtered and concentrated under reduced pressure to give the crude product. The desired Formula I compound {where R 1s -(CHj)4-] is obtained by purification of the crude product by use of HPLC.

While the prese-nt invention has been described with reference to the specific embodiments thereof, it should be understood by those skilled in the art that various changes may be made and equivalents may be substituted without departing from the trues spirit and scope of the invention. In addition, many modifications may be mnade to adapt a particular situation, material, composition of matter, process, process step or steps, to the objective spirit and scope of the present invention. All such modifications are intended to be within the scope of the claims appended heretos.

Claims

Bh WO 99/64042 PCT/US99/12728 -12._5- WHAT IS CLAIMED IS:

1. A multi-binding compound and salts thereof comprising 2 to 10 ligands, which may be the same or different, and w~hich are covalently attached to a linker or linkers which may be the same of differ<ent, at least one of said ligands comprising a ligand binding domain capab®e of binding to an alpha-1A adrenergic receptor.

2. The muiti-binding compound according to Claim 1 wherein at least two of the ligands comprises a ligand binding dormmain capable of binding to an alpha-1A adrenergic receptor.

3. A multibinding compound represented by Formula I: (WL) (X), Mm and pharmaceutically acceptable salts thereof, wherein: each L is a ligand that may be the samme or different at each occurrence: each X is a linker that may be the sa_me or different at each occurrence; p is an integer of from 2 to 10; and q is an integer of from 1 to 20; wherein each of said ligands comprises a ligand domain capable of binding to an alpha-1A adrenergic receptor, and wheere ¢ is less than p.

4. The multibinding compound of claim 3, wherein each of said ligands is capable of modulating the activity of the alpsha-1A adrenergic receptor.

S. The multibinding compound of claim 4, wherein each ligand capable of binding to the alpha-1A adrenergic receptor is independently selected from the group consisting of terazosin, prazosin, dox asosin, alfuzosin, tamsulosin, RS

100975, A-131701, L794-191, L757464, REC 15-2739, KMD-3213 and derivatives thereof.

6. The multibiending compound of claim 4, wherein each divalent linker X &s independently selec ted from a structure of Table 1.

7. The multibisnding compound of claim 6, wherein p is an integer of from 2 to 4, and q is less thar p.

8. A pharmaceutical composition comprising a pharmaceutically acceptable excipient and an effective amount of a multi-binding compound, or a pharmaceutically acceptable salt thereof, comprising 2 to 10 ligands which may be the same or different and which are covalently attached to a linker or linkers which may be the same or~ different, at least one of said ligands comprising a ligand domain capable of ®binding to one or more alpha-1A adrenergic receptors.

8. The pharmaceutical composition according te Claim 8 wherein said ligands comprising a ligancl domain capable of binding to one or more alpha-1A adrene=rgic receptors modulate benign prostatic hyperplasia and/or hypertension in mammals.

10. The pharmaaceutical composition according to Claim 9 wherein said liga nds are selected from the group consisting of terazosin, prazosin, doxasosin, alfuzoesin, tamsulosin, RS 100975, A-131701, L794-191, L.757464, REC 15-2739 and KMD- 3213, and derivatiwes thereof.

11. A pharmaceutical composition comprising a pharmaceutically acceptab le excipient and a thesrapeutically effective amount of one or more multibinding compounds represented by Formula I, LK, © and pharmaceutically acceptable salts thereof; wherein: each L is a ligand that may be the same or different at each occurrence, each X is a linker that may be the same or different at each occurrence; p is an integer of from 2 to 10; and - gis an integer of from 1 to 20; wherein each of said ligands comprises a ligand domain capable of binding to an alpha-1A adrenergic receptor, and where gq is less than p.

12. The pharmaceutical composition of claim 11, wherein said muitibinding compound is capable of modulating activity of the alpha—1A adrenergic receptor.

13. The pharmaceutical composition of claim 12, wherein each ligand is independently selected from the group consisting of terazosin, prazosin, doxasosin, alfuzosin, tamsulosin, RS 100975, A-131701, L794-191_ L757464, REC 15-2739, KMD-3213 and derivatives thereof.

14. The pharmaceutical composition of claim 12, wherein each linker X is independently selected from a structure of Table 1.

15. The pharmaceutical composition of claim 14, whexein p is an integer of from 2 to 4, and q 1s less than p.

16. A method of preparing a multibinding compound represented by formula I: wherein each L is a ligand that may be the same or different at each occurrence; X is a linker that may be the same or different at e ach occurrence; p is an integer of from 2 to 10; and q is an integer of from 1 to 20;

. wherein each of said ligands comprises a ligand domain capabl e of binding 10 an alpha-1A. adrenergic receptor, and where g is less than D. (a) pro viding at least p equivalents of a ligand L or precursors thereof and at least ¢ equivalents of linker or linkers X; and (b) covalently attaching said ligands to said linkers to produce a. mulubinding compound; or (b") covalently attaching said ligand precursors to said linkers an d completing the synthesis o£ said ligands thereupon. thereby to produce a mulubindimg compound.

17. The method of claim 16, wherein p is an integer of from 2 to 4, and gis less than p.

18. The met hod of claim 16, wherein each ligand is independently selected from the group consisting of terazosin, prazosin, doxasosin, alfuzosin. tamswlosin. RS 100975, A-131701, L794-191, L757464, REC 15-2739, KMD-3213 and derivatives thereof.

19. Use of a multi-binding compound, or a pharmaceutically acceptable salt thereof, comprising 2 to 10 ligands which may be the same or different and which are covalently attached to a linker or linkers which may be thes same or different, at least one of said ligands comprising a ligand domain capable of binding to one or more alpha-1A adrenergic receptors, in the manufacture of a medicament for weating benign prostatic hyperplasia and/or hypertension in a mammal. : AMENDED SHEET

20. Use of ore or more multibinding compounds represented by formula I, Lp0g or a pharmaceut®&cally acceptable salt thereof, wherein each L is a ligand that may be the same or different at each occuxrence; X is a linker that may be the same or different at each occurrences; p is an integer of from 2 to 10; and q is an integer of from 1 to 20; wherein each of said ligands comprises a ligand domain capable of binding to an alpima-1A adrenergic receptor, and where g is less than p, in the manufacture of a amedicament for modulating or alleviating benign prostatic hyperplasia and/o hypertension in a mammal.

21. Use of clabm 20, wherein p is an integer of from 2 to 4 and q is less than p. 135

22. Use accord ing to claim 20 wherein the ligand is selected from the group consisting of terazosin, prazosin, doxasosin, alfuzosin, tamsulosin, RS 1002975, A-131701, L794-1%91, L757464, REC 15-2739, KMD-3213 and derivatives thereof.

23. . A method for identifying multimeric ligand compounds possessing : 20 multibinding properties which method comprises: (a) identifying a ligand or a mixture of ligands wherein each liganci contains at least one reactive functionality and has a ligand binding domain capable of binding to an alphza-1A adrenergic receptor; (b) idenri®ying a library of linkers wherein each linker in said libraxy comprises at least tw Q functional groups having complementary reactivity 10 a t least * one of the reactive fimnctional groups of the ligand; AMENDED SHEET

(crm preparing a multimeric ligand compound library by combining at least two stoichiometric equivalents of the ligand or mixture of ligands identified in (a) with the library of linkers identified in (b) under conditions wherein the compleme=ntary functional groups react to form a covalent linkage between said linker andl at least two of said ligands; and (d» assaying the multimeric ligand compounds produced in the library prepared #in (c) above to identify multimeric ligand compounds possessing ) multibind®ng properties.

24. A zmethod for identifying multimeric ligand compounds possessing multibind®ng properties which method comprises: (a) identifying a library of ligands wherein each ligand contains at least one reactive functionality and has a ligand binding domain capable of binding to an alpha-1A =adrenergic receptor; (b)- identifying a linker or mixture of linkers wiaerein each linker comprises at least two functional groups having complementary reactivity to at least one of the reactive functional groups of the ligand; (c) preparing a multimeric ligand compound library by combining at least two Stoichiometric equivalents of the library of ligands identified in (a) with the linker or mixture of linkers identified in (b) under conditions wherein the compleme=ntary functional groups react to form a covalent linkage between said linker and_ at least two of said ligands; and (dm assaying the multimeric ligand compounds produced in the library prepared &n (c) above to identify multimeric ligand compowinds possessing multibind®ng properties.

25. Time method according to Claim 23 or 24 wherein the preparation of the multimeric ligand compound library is achieved by either the sequential or concurrent combination of the two or more stoichiometric equivalents of the ligands identified in (a) with the linkers identified in (b).

26. The method according to Claim 25 wherein the mu_ltimeric ligand comp ounds comprising the multimeric ligand compound li ‘brary are dimeric.

27. The method according to Claim 26 wherein the dimeric ligand compounds comprising the dimeric ligand compound library are heterodimeric.

28. The method according to Claim 27 wherein the heterodimeric ligand compeound library is prepared by sequential addition of a first and second ligand.

29. The method according to Claim 23 or 24 wherein, prior to procedure (d), each rmember of the multimeric ligand compound library is isolated from the librar=y.

30. The method according to Claim 29 wherein each m ember of the library is isolated by preparative liquid chromatography mass spectrometry (LCMS).

31. The method according to Claim 23 or Claim 24 wherein the linker or linker=s employed are selected from the group comprising flexible linkers, rigid linkerss, hydrophobic linkers, hydrophilic linkers, linkers o=f different geometry, acidic linkers, basic linkers, linkers of different polarizatio-n and/or polarizability and armphiphilic linkers.

32. The method according to Claim 31 wherein the linkzers comprise linkers of different chain length and/or having different complementary reactive groups.

33. The method according to Claim 32 wherein the linkkers are selected to have different linker lengths ranging from about 2 to 100A.

34. The method according to Claim 23 or 24 wherein tthe ligand or mixture of ligands is selected to have reactive functionality at differemmt sites on said ligands.

35. The method according to Claim 34 wherein said rezactive functionality is selected from the group consisting of carboxylic acids, carboxylic acid halides, carboxyl esters, amines, halides, pseudohalides, isocyanates, vinyl unsaturation, ketone=s, aldehydes, thiols, alcohols, anhydrides, boronates , and precursors thereof where=in the reactive functionality on the ligand is selected to be complementary to at least one of the reactive groups on the linker so that a covalent linkage can be forme d between the linker and the ligand.

36. The method according to Claim 23 or Claim 24 wherein the multimeric ligand compound library comprises homomeric ligand com pounds.

37. The method according to Claim 23 or Claim 24 wherein the multimeric ligand compound library comprises heteromeric ligand conrapounds.

38. A library of multimeric ligand compounds which may possess multivalent proper-ties which library is prepared by the method comprising: (a) identifying a ligand or a mixture of ligands vwherein each ligand contaims at least one reactive functionality and has a ligand binding domain capable of bincling to an alpha-1A adrenergic receptor; (b) identifying a library of linkers wherein each linker in said library compr ises at least two functional groups having complemeratary reactivity to at least one of the reactive functional groups of the ligand; and

© WO 99/64042 PCT/USY9/12728 (c) preparing a multimeric ligand compound library by combining at least two stoischiometric equivalents of the ligand or mixture of ligands identified in (a) with the l#brary of linkers identified in (b) under conditions wh erein the complementa-ry functional groups react to form a covalent linkage between said linker and at least two of said ligands.

39. A library of multimeric ligand compounds which may poss ess multivalent properties which library is prepared by the method comprising: (@) identifying a library of ligands wherein each ligand contains at least one reactive Sunctionality and has a ligand binding domain capable of binding to an alpha-1A adreenergic receptor; (b) identifying a linker or mixture of linkers wherein each linker comprises at least two functional groups having complementary reactivity to at least one of the reactive functional groups of the ligand; and (©) preparing a multimeric ligand compound library by combining at least two stoichiometric equivalents of the library of ligands identi fied in (a) with the linker or mixture of linkers identified in (b) under conditions wherein the complementary functional groups react to form a covalent linkage between said linker and at least two of said ligands.

40. The l3brary according to Claim 38 or Claim 39 wherein the linker or linkers employed are selected from the group comprising flexible linkers , rigid linkers, hydrophobic linkers, hydrophilic linkers, linkers of different geormetry, acidic linkers, basic linkers, linkers of different polarization and/or polarizability and amphiphilic Jinkers.

41. The Library according to Claim 40 wherein the linkers cormprise linkers of different cha. in length and/or having different complementary reactive groups.

42. The library according to Claim 41 wherein the linkers are selected to have different linker lengths ranging from about 2 to 10»0A.

43. The library according to Claim 38 or 39 wherein the ligand or mixture of ligands is selected to have reactive functionality at different sites on said ligands.

44. The library according to Claim 43 wherein said reactive functionality is selected from the group consisting of carboxylic ac ids, carboxylic acid halides, carboxyl esters, amines, halides, pseudohalides, isocyanates, vinyl unsaturation, ketones, aldehydes, thiols, alcohols, anhydrides, boOronates, and precursors thereof wherein the reactive functionality on the ligand is s lected to be complementary to at least one of the reactive groups on the linker so that a covalent linkage can be formed between the linker and the ligand.

45. The library according to Claim 38 or Claim 39 wherein the multimeric ligand compound library comprises homomeric ligand compounds.

46. The library according to Claim 38 or Claim 39 wherein the multimeric ligand compound library comprises heteromeric ligand compounds.

47. An iterative method for identifying multimeric ligand compounds possessing multibinding properties which method comprises: (a) preparing a first collection or iteratio n of multimeric compounds which is prepared by contacting at least two stoichiometric equivalents of the ligand or mixture of ligands which target an alpha-1A adrenergic receptor with a linker or mixture of linkers wherein said ligand or mixture of ligands comprises at least one : reactive functionality and has a ligand binding domamin capable of binding to an alpha-1A adrenergic receptor, and said linker or mixture of linkers comprises at least two functional groups having complementary reactivity to at least one of the

) © WO 99/64042 [amp reactive functional groups of the ligand wherein said contacting is conducted under conditions wherein the complementa ty functional groups react to form a covalent linkage between said linker and at le ast two of said ligands; (b) assaying said first col Jection or iteration of multimeric compounds to assess which if any of said multimer ic compounds possess multibinding properties; (c) repeating the process of (a) and (b) above until at least one multimeric compound is found to possess multibinding properties: d) evaluating what molecular constraints imparted or are consistent with imparting multibinding properties to the multimeric compound or compounds found in the first iteration recited in (a)- (c) above: (© creating a second collection or iteration of multimeric compounds which elaborates upon the particular molecular constraints imparting multibinding properties to the multimeric compound or compounds found in said first iteration: H evaluating what molecular constraints imparted or are consistent with imparting enhanced multibindimng properties to the multimeric compound or compounds found in the second coll ection or iteration recited in (e) above; (g) optionally repeating Steps (e) and (f) to further elaborate upon said molecular constraints.

48. The method according to Cl aim 47 wherein steps (e) and (f) are repeated from 2-50 times.

49. The method according to CLaim 47 wherein steps (e) and (f) are repeated from 5-50 times.

E

50. A substance or compos ition for use in a method for treating benign prostatic hyperplasia and/or hypertension in a mammal, said substance or composition comprising a multi-binding compound, or a pharmaceutically acceptable salt thereof, comprising 2 to 10 ligands which may be the same or h) different and which are covalently attached to a linker or linkers which may be the same or different, at least one of said ligands comprising a ligand domain capable of binding to one or more alpha-1A adrenergic receptors, and said method comprising administering to said mammal an effective amount of said substance or composition.

51. A substance or compo sition for use in a method for modulating or alleviating benign prostatic hyperplasia and/or hypertension in a mammal, said substance or composition comprising one or more multibinding compounds represented by formula I, LpX)q OD and pharmaceutically acceptable salts thereof, wherein each L is a ligand that may be the same or different at each occurrence; X is a Urker that may= be the same or different at each occurrence; p is an integer of from 2 to 10; and q is an integer of frorm 1 to 20; wherein each of said ligands comprises a ligand domain capable of binding to an alpha-1A adrenergic receptor, and where ¢ is less than p, and said method comprising administering to a mammal in need of such treatment a therapeutically effective amount of said substance or composition.

52. A substance or composition for use in a method of treatment according to claim 51, wherein p is an. integer of from 2 to 4 and q is less than p.

53. A substance or composition for use in a method of treatment according to claim 50 wherein the ligand is selected from the group consisting of AMENDED SHEET é t terazosin, prazosin, doxasosin, alfuzosin, tarmsulosin, RS 100975, A-131701, 1794-191, L757464, REC 15-2739, KMD-3 213 and derivatives thereof.

54. A compound according to claim 1 or claim 3, substantially as herein described and illustrated.

55. A composition according to claim 8 or claim 11, substantially as herein described and illustrated.

56. A method according to claim 16, sub stantially as herein described and illustrated.

57. Use according to claim 19 or claim 2 O, substantially as herein described and illustrated.

58. A method according to claim 23, claimm 24 or claim 47, substantially as herein described and illustrated.

59. A library according to claim 38 or claim 39, substantially as herein described and illustrated.

60. A substance or composition for use in a method of treatment according to claim 50 or claim 51, substantially as herein described and illustrated.

61. A new compound, a new composition, a new method of preparing a compound, new use of a multi-binding compowund as claimed in any one of claims 1 - 7, a new method for identifying compounds, a new library, or a substance or composition for a new use in a method of treatment, substantially as herein described. AMENDED SH EET