LV10425B - Fibrinogen receptor antagonists - Google Patents

Abstract

Fibrinogen receptor antagonists with formula<IMAGE>are applied to inhibit fibrinogen attraction to haematoceles as well as the haematocele aggregation.

Description

LV10425

TITLE OF THE INVENTION FIBRINOGEN RECEPTOR ANTAGONISTS

FIELD OF THE INVENTION

This invention relates to the discovery of fibrinogen receptor antagonists of Formula I for use in inhibiting the binding of fibrinogen to blood platelets and inhibiting the aggregation of blood platelets when administered to mammals, preferably humāns.

BACKGROUND OF THE INVENTION

The interaction of platelets with the coagulation and fibrinolytic systems in the maintenance of hemostasis may become pathogenic, 2 requiring prevention and treatment. The fibrinogen receptor antagonists of Formula I are useful in treating various diseases related to platelet aggregation and fibrin formation.

An interest in platelet inhibitors has reemerged as a result of a better understanding of the role of platelets and thrombosis in the pathogenesis of vascular disease, including unstable angīna, acute myocardial infarction and stroke.

Platelets are cell-like anucleated fragments, found in the blood of ali mammais, which participate in blood coagulation. Fibrinogen is a glycoprotein present as a normai component of blood plasma. Fibrinogen participates in platelet aggregation and fibrin formation in the blood clotting mechanism. Platelets are deposited at sites of vascular injury where multiple physiological agonists act to initiate platelet aggregation culminating in the formation of a platelet plug to minimizē blood loss. If the platelet plug occurs in the lumen of a blood vessel, normai blood flow is impaired.

Platelet membrane receptors are essential in the process of platelet adhesion and aggregation. Interaction of fibrinogen with a receptor on the platelet membrane complex Ilb/IIIa is known to be essential for normai platelet function.

Zimmerman et al., U.S. Patent No. 4,683,291, describes peptides having utility in the study of fibrinogen-platelet, platelet-platelet, and cell-cell interactions. The peptides are described as having utility where it is desirable to retard or prevent formation of a thrombus or clot in the blood. - 3 - LV10425

Pierschbacher et al. , U.S. Patenti No. 4,589,881, describes the sequence of an 11.5 kDal polypeptide fragment of fibronectin which embodies the cell-attachment-promoting activity of fibronectin.

Ruoslahti et al., U.S. Patent No. 4,614,517, describes tetrapeptides which alter cell-attachment activity of celis to various substrates. Figurē 1 lists the polypeptides that were synthesized by Ruoslahti et al. in &quot;determining the smallest peptide exhibiting celi attachment activity.&quot; Ruoslahti et al., U.S. Patent No. 4,578,079, describes similar tetrapeptides.

Pierschbacher et al., Proc. Nati. Acad Sci. USA. Vol. 81, pp.5985-5988, October, 1984, describe variants of the celi recoglīition site of fibronectin that retain attachment-promoting activity. Pierschbacher et. al. further assayed the celi attachment-promoting activities of a number of structures closely resembling the Arg-Gly-Asp-Ser peptide, and found &quot;that the arginine, glycine, and aspartate residues cannot be replaced even with closely related amino acids, but that several amino acids can replace serine vithout loss of activity.&quot;

Ruoslahti et al., Science. Vol. 238, pp. 491-497, October 23, 1987, discuss celi adhesion proteīns. They specifically State that &quot;elucidation of the amino acid sequence of the cell-attachment domain in fibronectin and its duplication with synthetic peptides establish the sequence Arg-Gly-Asp (RGD) as the essential structure recognized by celis in fibronectin.&quot; 4

Cheresh, Proc. Nati. Acad. Sci. USA. Vol. 84, pp. 6471-6475, September 1987, describes the Arg-Gly-Asp-directed adhesion receptor involved in attachment to fibrinogen and the von Willebrand Factor. Ādams et al., U. S. Patent No. 4,857,508, describes tetrapeptides which inhibit platelet aggregation and the formation of a thrombus.

It is, therefore, an object of the present invention to provide fibrinogen receptor antagonists for use in inhibiting the binding of fibrinogen to blood platelets and inhibiting the aggregation of blood platelets. Another aspect of the present invention is to provide novel fibrinogen receptor &quot;antagonist compounds. Other objects of the present invention are to provide methods of inhibiting the binding of fibrinogen to blood platelets and inhibiting the aggregation of blood platelets, through the administration of novel fibrinogen receptor antagonist compounds. The above and other objects are accomplished by the present invention in the manner described below.

SUMMARY OF THE INVENTION

The present invention provides fibrinogen receptor antagonist compounds of the formula:

0 R

X-&lt; CH2)m-Y-(CH2)ķ-C-NH-CH-CE-Z

Rl

I - 5 - LV10425 for use in inhibiting the binding of fibrinogen to blood platelets and for inhibiting the aggregation of blood platelets. The above-mentioned compounds can be used in a method of acting upon a fibrinogen receptor which comprises administering a therapeu-tically effective but non-toxic amount of.such compound to a mammai, preferably a human. A pharmaceutical composition comprising a pharmaceu-tically acceptable carrier and, dispersed therein, an effective but non-toxic amount of such compound is another feature of this invention.

DĒTAILED DESCRIPTION OF THE INVENTION

Fibrinogen receptor antagonist compounds of Formula I are useful in a method of inhibiting the binding of fibrinogen to blood platelets and for inhibiting the aggregation of blood platelets. Fibrinogen receptor antagonists of this invention are illustrated by compounds having the formula:

0 R

X-(CH2)m-Y-(CH2)k-C-NH-CH-CH-Z

Rl

I 6 wherein: X is

NH NH NH -NR7R3, -NH-'Č-NR7R3, -C-NR7R3, r7-c-nh-

0

II where D= -C-, -S(0)q, or -0-;

where A=N and B= -CH2-, or A= -CH- and B= NR7 ; 30 - 7 - LV10425 Y is

Ο Ο Ζ is -C09R2; -Ρ-ΟΗ -Ρ-ΟΗ OR2 ; RlO ; where R^-® is Cļ_g alkyl, aryl, aryl Cļ_g alkyl; u is -CH-, -C-, or -N-; v is -CH-, -C-, or -N-; R and R^ are independently hydrogen, aryl, wherein aryl is defined as a mono- or polycyclic aromatic system comprised of 5 or 6 membered rings containing 0, 1, 2, 3, or 4 heteroatoms selected from nitrogen, oxygen and sulfur; and phenylene, either unsubstituted or substituted, with one or more groups selected from hydroxyl, fluoro, chloro, bromo, iodo, cyano, trifluoromethyl, Cļ_3 alkoxy, Cļ_5 alkylcarbonyloxy, Cļ_5 alkoxycarbonyl, Cļ_5 alkyl, aminoCļ_5 alkyl, hydroxycarbonylCg_5 alkyl, or hydroxycarbonylCļ_5 alkoxy, 8 CQ_^alkyl, either unsubstituted or substituted, with one or more groups selected from fluoro, chloro, bromo, iodo, hydroxyl, Cļ_5alkylcarbonyl-(CQ_galkyl)amino, arylCļ_5alkyl carbonyl(CQ_galkyl)amino, aryloxy, C1_10alkoxy, Cļ_5alkoxycarbonyl, CQ_5alkylaminocarbonyl, Cļ_5 alkyl-carbonyloxy, C3_gcycloalkyl, aryl, oxo, amino, Cļ_galkyl, Cļ_3alkylamino, aminoCļ_3alkyl,arylCQ_5alkylamino-carbonyl, phenylCļ_3 alkylamino, amino-carbonylCQ_4alkyl, Cļ_galkyl-sulfonyl (CQ_galkyl)amino, arylCo_io- alkylsulfonyl(CQ_galkyl)-alkylsulfonyl, Co_8alkylsulfonyl, hydroxycarbonyl-CQ_5alkyl, C]__galkyloxycarbonyl-(CQ_galkyl)amino, arylCQ_ļQalkyloxy-carbonyl(CQ_galkyl)amino, CQ_g alkylaminocarbonyl(CQ_g alkyl)amino, arylCQ_galkylaminocarbonyl(Cg_8” alkyl)amino, CQ_galkylaminocarbonyloxy, arylCQ_ļQalkylaminocarbonyloxy * ^0-8-alkylaminosulfonyl(Cq_3 alkyl)amino, arylCo_8al^ylaminosulf onyl^(-0-8al^y1) amino, CQ_galkylaminosulfonyl, or arylCo_8aikylaminosulfonyl; provided that the carbon atom to which R or is attached bear only one heteroatom; - 9 - LV10425 is hydrogen,

Ci_i2alkyl. unsubstituted or substituted, with one or more Cļ_6alkyl groups, 0 R9 0 CHo CHo 0 -CH20C-R9, -CHOCR9 or -C-0-C-R9 where R9 = Cļ_6alkyl, branched or unbranched, or phenyl, and wherein R9, when appearing more than once, can be same or different; R7, r3 and are independently • hydrogen,

Ci_i2 alkyl, unsubstituted or substituted, with one or more Cļ_^ alkyl groups, arylCQ_4alkyl, or cyano

provided that wheft R7 and R3 are independently cyano, X NH NH is -NH-C-NR7R3 or -C-NR7R3; k is 1-4; m is 1-4; p is 1-6; q is 0-2; or the pharmaceutically optical isomers thereof acceptable salts thereof, or 10

More preferred compounds of the present invention have the following formula:

0 R X-(CH2)m-Y-(CH2)k-C-NH-CH-CH-C02R2

Rl

I wherein:

NH X is -NR7R3, -C-NR7R3,

r7_N Cch2). -r7-n R4 Λ

N

H,N

, or

H2N 0

II where D = -C-, -S(0)q-, or -0-; Z is C02r2 Y is

- 11 - LV10425 wherein n is 1, 2 or 3; R and are independently chosen from phenyl, thiophene, imidazole, naphthyl, indole, indazole, thionaphthene, either unsubstituted or substituted, with hydroxy, halogen, hydroxy-carbonyl Cq_5 alkyl, Cļ_3alkyl, either unsubstituted or substituted, with one or more groups selected form aryl, aryloxy, Cļ_]_Q alkoxy, Cq_5 alkylaminocarbonyl, arylCQ_5 alkylaminocarbonyl, hydrogen, CQ_6alkyl either unsubstituted or substituted, with one or more groups selected from halogen, hydroxyl, C1_6alkylsulfonylamino, aryl CQ_5alkylsulfonylamino, Cļ_0alkylsulfonyl, arylCQ_5alkylsulfonyl, Cļ_5alkylcarbonylamino, arylCļ_5alkylcarbonylamino, aryloxy, Cļ_ļQ alkoxy, Cļ_5alkoxycarbonyl, CQ_5alkylamino-carbonyl, Cļ_5alkylcarbonyloxy, C3_scycloalkyl, aryl, oxo, amino, Cļ_0alkyl, Cļ_3alkylamino, aminoCļ_3 alkyl, arylCQ_5 alkylaminocarbonyl, phenylCļ_3 alkylamino, aminocarbonylCQ_4 alkyl, or hydroxycarbonylCQ_3 alkyl, provided that the carbon atom to which R or R* is attached bear only one heteroatom, R2 is hydrogen,

Ci_iz alkyl, unsubstituted or substituted, with one or more C]__£alkyl groups, 12 0 R9 Ο CH? CH? 0 II η ' “ Q N / &quot; d -CH20C-R9, -CH0CR9 or -C-0-C-R9 where R9 = C^_^alkyl, branched or unbranched, or phenyl, and vherein R9, when appearing more than once, can be the sarae or different; R7, R3 and R^ are independently hydrogen, or Cļ_3alkyl, unsubstituted or substituted, with one or more Cļ_^alkyl groups; k is 1-4; m is 1-4; q is 0 or 2; p is 1-3; ~ or the pharmaceutically acceptable salts thereof, or optical isomers thereof.

Most preferred compounds of the present invention have the following formula:

0 R II i n

X&quot;(CH2)m-Y-CH2-C-NH-CH-CH-C02RZ

Rl

- 13 - LV10425 -NR7R3; 0 ll where D = -0-, -S-, or -C-, Z is COļR2 Y is

R

o

vherein n is 1, 2 or 3; R and R^ are independently chosen from phenyl, thiophene, imidazole, naphthyl, indole, indazole, thionaphthene, either unsubstituted or substituted, with hydroxy, halogen, hydroxy-carbonyl Cq_5 alkyl, C]__3alkyl, either unsubstituted or substituted, with one or more groups selected form aryl, aryloxy, C]__ļQ alkoxy, C0_5 alkylaminocarbonyl, arylCQ_5 alkylaminocarbonyl, hydrogen, CQ_^alkyl, either unsubstituted or substituted, with one or more groups selected from halogen, hydroxyl, Cļ_3alkylcarbonylamino, arylCļ_5 alkylcarbonylamino, aryloxy, Cļ_ļQ alkoxy, Cļ_5alkoxycarbonyl, Co_5alkylaminocarbonyl, 14 - Cļ_5alkylcarbonyloxy, C3_gcycloalkyl, aryl, oxo, amino, Cļ_galkyl, C1_3alkylami.no, aminoCļ_3 alkyl, arylC0_5 alkylaminocarbonyl, phenylCļ_3 alkylamino, aminocarbonylCQ_^ alkyl, or hydroxycarbonylCQ_5 alkyl, provided that the carbon atom to which R or is attached bear only one heteroatom; R2 is hydrogen; R^, R2 and R^ are hydrogen; m is 1-4; p is 2-4; or the pharmaceutically acceptable salts thereof, or optical isomers thereof.

This invention includes the following abbreviation designations; Bn, benzyl; NMM, N-methylmorpholine; HOBt, l-hydroxybenzotriazole; EDC, l-(3-dimethylaminopropyl)-3-ethylcarbodiimide-hydrochloride; DMF, dimethylformamide; BOC, tert-butyloxycarbonyl; pTSA, para-toluenesulfonic acid; DMS, dimethylsulfide; TFA, trifluoroacetic acid; THF, tetrahydrofuran; TBDMS, tert-butyldi-methylsilyl.

The pharmaceutically acceptable salts of the compounds of Formula I include the conventional non-toxic salts or the quarternary ammonium salts of the compounds of Formula I formed, e.g., from non-toxic inorganic or organic acids. For example, such conventional non-toxic salts include those derived from inorganic acids such as hydrochloric, - 15 - LV10425 hydrobromic, sulfuric, sulfamic, phosphoric, nitric and the like; and the salts prepared from organic acids such as acetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamoic, maleic, hydroxyraaleic, phenylacetic, glutamic, benzoic, salicylic, sulfanilic, 2-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, isethionic, and the like.

The pharmaceutically acceptable salts of the present invention can be synthesized from the compounds of Formula I which contain a basie or acidic moiety by conventional Chemical methods. Generally, the salts are prepared by reacting the free base or acid with stoiehiometric amounts or with an excess of the desired salt-forming inorganic or organic acid or base in a suitable solvent or various combinations of solvents.

The pharmaceutically acceptable salts of the acids of Formula I are also readily prepared by conventional procedures such as treating an acid of Formula I with an appropriate amount of a base, such as an alkali or alkaline earth mētai hydroxide e.g. sodium, potassium, lithium, calcium, or magnesium, or an organic base such as an amine, e.g., dibenzyl-ethylenediamine, trimethylamine, piperidine, pyrrolidine, benzylamine and the like, or a quaternary ammonium hydroxide such as tetramethylammonium hydroxide and the like.

The compounds of Formula I are useful in inhibiting the binding of fibrinogen to blood platelets, inhibiting aggregation of blood 16 platelets, treatment of thrombus formation or embolus formation, in the prevention of thrombus formation or embolus formation, and in the mediation of cell-cell fusion events, such as sperm-egg fusion, resulting in mammalian fertilization inhibition. These compounds are useful as pharmaceutical aģents for mammals, especially for humāns. The compounds of this invention may be administered to patients where prevention of thrombosis by inhibiting binding of fibrinogen to the platelet membrane glycoprotein complex Ilb/IIIa receptor is desired. Compounds of this invention may also be used to prevent or modulate the progress of myocardial infarction, unstable angīna and thrombotic stroke, in either acute or chronic settings. In addition, they may be useful in surgery on peripheral arteries (arterial grafts, carotid endarterectomy) and in cardiovascular surgery where manipulation of arteries and orgāns, and/or the interaction of platelets with artificial surfaces, leads to platelet aggregation and consumption. The aggregated platelets may form thrombi and thromboemboli. Compounds of this invention may be administered to surgical patients to prevent the formation of thrombi and thromboemboli.

Extracorporeal circulation is routinely used for cardiovascular surgery in order to oxygenate blood. Platelets adhere to surfaces of the extracorporeal Circuit. Adhesion is dependent on the interaction between GPIĪb/IIIa on the platelet raembranes and fibrinogen adsorbed to the surface of the Circuit. (Gluszko et al., Amer. J. Phvsiol^ 1987, 252:H, pp 615-621). Platelets released from artificial surfaces show impaired - 17 - LV10425 heraostatic function. Compounds of this invention may be administered to prevent adhesion.

Other applications of these compounds include prevention of platelet thrombosis, thromboembolism, reocclusion, and restenosis during and after thrombolytic therapy and prevention of platelet thrombosis, thromboembolism, reocclusion and restenosis after angioplasty of coronary and other arteries and after coronary artery bypass procedures.

The compounds of Formula I may be administered to mammals, preferably in combination with pharmaceutically-acceptable carriers or diluents, optionally with known adjuvants such as alum, in a pharmaceutical composition which is non-toxic and in a therapeutically effective amount, according to Standard pharmaceutical practice. The compounds can be administered orally or parenterally, including intravenous, intramuscular, intraperitoneal, trans-dermal, subcutaneous and topical administration.

For oral use of a fibrinogen receptor antagonist according to this invention, the selected compounds may be administered, for example, in the form of tablets or capsules, or as an aqueous solution or suspension. In the case of tablets for oral use, carriers which are commonly used include lactose and corn starch, and lubricating aģents, such as magnesium stearate, are commonly added. For oral administration in capsule form, useful diluents include lactose and dried corn starch. When aqueous suspensions are required for oral use, the active 18 ingredient is combined with emulsifying and suspending aģents. If desired, certain sweetening and/or flavoring aģents may be added.

For intramuscular, intrapertioneal, subcutaneous and intravenous use, sterile Solutions of the active ingredient are usually prepared, and the pH of the Solutions should be suitably adjusted and buffered. For intravenous use, the total concentration of solutes should be controlled in order to render the preparation isotonic.

The present invention also encompasses a pharmaceutical composition useful in the treatment and prevention of diseases related to platelet aggregation, fibrin formation, and thrombus and embolus formation, comprising the administration of a therapeutically effective but non-toxic amount of the compounds of Formula I, with or without pharmaceutically acceptable carriers or diluents.

Compositions of this invention include fibrinogen receptor antagonist compounds of this invention in. combination with pharmacologically acceptable carriers, e.g. saline, at a pH Ievel e.g. 7.4, suitable for achieving inhibition of platelet aggregation. The compositions may also be combined with anticoagulants such as heparin or warfarin.

The compositions may also be combined with thrombolytic aģents such as plasrainogen activators or streptokinase in order to inhibit platelet aggregation in more a'cute settings. The composition may further be combined with antiplatelet aģents such as aspirin. The compositions are soluble in an aqueous medium, and may therefore be effectively administered in solution. - 19 - LV10425

When a corapound according to Formula I is used as a fibrinogen receptor antagonist in a human subject, the daily dosage will normally be determined by the prescribing physician with the dosage general.ly varying according to the age, weight, and response of the individual patient, as well as the severity of the patients symptoms.

In one exemplary application, a suitable amount of compound is administered orally to a heart attack victim before or subsequent to angioplasty. Administration occurs before or subsequent to angioplasty, and is in an amount sufficient to inhibit platelet aggregation, e.g. an amount which achieves a steady State plasma concentration of between about 0.01-100 μΜ preferably betveen about 0.01-10 μΜ.

The present invention also includes a pharmaceutical composition comprising compounds of the present invention in combination with tissue type plasminogen activator or streptokinase. The invention also includes a method for promoting thrombolysis and preventing reocclusion in a patient which comprises administering to the patient an effective amount of compositions of the invention.

The present invention provides a method of inhibiting the binding of fibrinogen to blood platelets, inhibiting aggregation of blood platelets, treating thrombus formation or embolus formation, and in preventing thrombus formation or embolus formation in a mammai, comprising the administration of a therapeutically effective but non-toxic amount of the compounds of this invention, with or without pharmaceutically acceptable carriers or diluents. 20

The present invention stili further provides a method of inhibiting the binding of fibrinogen to blood platelets, inhibiting aggregation of blood platelets, treating thrombus formation or embolus formation, and in preventing thrombus formation or embolus formation in a mammai, comprising the administration of a therapeutically effective but non-toxic amounts of the compounds of this invention in combination with thrombolytic aģents, such as tissue plasminogen activators or streptokinase, anticoagulants such as heparin or warfarin, or antiplatelet aģents such as aspirin, with or without pharmaceutically acceptable carriers or diluents. - 21 - LV10425

The following compounds and associated IC^g values (concentrations which inhibit aggregation by 507» relative to a control lacking the compound) are exemplary of the invention.

30 22 22 ΗΝ

sch3 ο ^ ν^Λ^°°η ch„ - 23 - 5 LV10425 IC50

20

30 24

ΗΝ

- 25 - LV10425

20

8. 1 MM 25 N. ΗΝ·

O

0. 84 mM 30 5 26 10 15

IC50

Ο. 27 /iM 0. 028 ϋΜ

30 27

ΗΝ

Ο&quot; Ο Η Ν. ςΗ3 co2h ch3 I-1 0 ! 28 Ο ΗΝ Ν N^C02H , ΗΝ 1 ο CH3 1 ,co2h . ο Η ΗΝ Ο .Ν. &gt;ο3η . ο Η - 29 - LV10425

Compounds of the invention can be prepared according to one of the following general procedures: SCiīSMS ,1

An aryl or alkyl alcohol is protected if needed and is oxidized to the corresponding aldehyde via oxalyl chloride/DMSO, pyridinium chlorochromate or similar reaģent. Typically, these oxidations are carried out in halocarbon solvents such as CH2CI2 at -78® to room temperature for 0.5-6 hrs. Olefination of the resulting aldehyde with a carboalkoxymethylene phosphorane proceeds to give the desired unsaturated ester. Typically, this reaction is run in halocarbon solvents such as CH2CI2 at 0-60° for 1-18 hours. Treatment of this ester with a chiral araine such as R-(+)-a-methylbenzylamine provides a diastereomeric mixture of amines that can be separated typically by column chromatography. SCEEME 2

An aminoalkanol is protected on N with a BOC, or similarly suitable group such as CBZ, FMOC, etc., and converted to the alkyl halide, typically the iodide. PI13P and iodine are typically used for this purpose and the reaction is run in aromatic solvents such as benzene or toluene in halocarbons such as CH2CI2 at 0-50° for 5 minūtes to 10 hrs. Formation of an alkyl azide is effected by treatment with NaN3 or similar azide transfer aģent, and this azide is catalytically reduced to the corresponding amine in alcohol solvents (EtOH, CH3OH) at room temperature over the course of 0.5-10 hrs. 30 SCHEME 3

Alkylation of cyclic amide or urea intermediates occurs in the presence of a suitable base, such as LiN(TMS&gt;2 or simple amines such as EtN(i-Pr)2 and an appropriate alkylating aģent such as an alkyl in alkyl iodide or a bromide such as ethyl bromoacetate. Disubstitution in the intermediate is effected by repetition of the deprotonation/alkylation seguence. When an allyl reaģent is used to alkylate, subsequent oxidation with RuCl3/NaI04 or KMn04 or other suitable reaģent provides the termiņai carboxylic acid. This acid may then be coupled via an amide bond with C-terminal chiral or achiral amines to provide further intermediates. Typically, coupling reactions are carried out in DMF or halocarbon solvents such as CH2CI2 utilizing Standard reaģents such as DCC, EDC, i-butyl chloroformate and the like. Final de-protection of intermediates typically involves base catalyzed hydrolysis of esters and acid catalyzed N-deprotection.

The compounds of Formula I are prepared according to the reaction schemes set forth below. Compound 1, 3-indolepropanol, is commercially available from Aldrich Chemical. - 31 - LV10425 SCHEME 1

Ac20, 4-dirret hylarrino-pyrldine, CH2C12, 1,8-diazabicyclo — [ 5. 4. 0] undec-7-ene

a) oxalyl chloride, DhSO, CH2C12, NEt3, -78° to 0 C b) Ph3P=CHC02 Et

Ac 5 32 SCHEME 1 cont'd

20% Pd(0H)a/C ECOH, H2 o h2n 7a

Compound 8 is commercially available from American Tokyo Kansei, Inc.

- 33 - HN Ή 8 SCHEME 2 boc2o, dmf

9 LV10425

BOC-N iodlna, Ph3P. irn idazole, benzene

NaN3, DKSO 70°C

BOC-N

•I 1 1 1 0 1 0% Pd/C, achanol, H2

BOC-N NH2 1 2 34SCHEME 3 Ο

HN NH \-J 1 3 1 0 O a) DMF, L1NCI^E)2 then allyl brorride b) LiN(T1*E) 2, then boc- n V^-&quot;1

O

- 35 -LV10425

SCHEME 3 cont'd BOC

BOC

TFA, CH2C12, anisole, -15°C

5 36 BOC-N 12 10 15 20 25 SCHEME 4 a) 4-chlorobutyryl chlorlde .NHj NEt(l-Pr)j, CHjCN, 0°C b) DMF. 0°C. NaN(Tf-S), THF, LDA, -7Θ °C, allyl brorrlde

30 - 37 -LV10425 SCHEME 4 cont'd

1 N NaOH, echanol

23 38 25 SCHEME 4 cont'd23

30 - 39 - SCHEME 5 LV10425

5-chlorovaleryl chloride, DhF, NMM λ- NH ''/'V&quot;C1 26

THF, NaN(ThE)2 -78°C to 0°C 0

THF. NaNCTm)^. -78eC 0

BOC-N then 10 28 21_

RuC13, CC14, CH3CN, H20, NaI04

BOC-N

COjH EDC, HOBT, NEt3, DMF, 7 a 30 29 40 SCHEME 5 cont'd

1N NaOH, ethanol

TFA, CH2C12, anisole

32 - 41 -5 SC5EME 6 LV10425 a) 4-chlorobucyryl chloride,

| DMF, NMM H2N^^Ph I ~

b) NaNflTS^, THF, -7B°C co 25°C O Ph 34 10 ir5

O

'Ph “— 35b -tCHŠH· Li, THF. -78°C LDA. -78°C 1 0 THF, , C han 2 0

25 30 42SflffEME 6 cont'd

THF, NaNCThE)z. -7a°C, Chen ethyl brorroacetate

BOC- o

BOC-N

Ņ 'C02H

O 38a N^COjH 36b 43 -LV10425 SCHEME 6 cont'd 38a EDC, HDBT, NEt3, DMF. 7a

BOC-N

—' O 39a 1 N NaOH. echanol

BOC-N

40a

TFA. CH2C12, anisole, -15°C

HN

41 a 44 SCHEME 6 cont'd 38b EDC, HOBT, NEC 3, DMF, 7 a

1N NaOH. echartol

TFfl, CH2C12, anlsole, -15°C

LV 10425

Svern oxidation

46 SCHEME 7 (continued)

- 47 - LV 10425 td d* S..M&lt;

z r •H s o z B o n X o o rt o u

r- m SCHEME 7 (continued) £ bl ifl m I_&gt; U3 m o&quot; £

o B

U£ CJ ai£ u m Ui U

a . £ — o z s Γ» o £ B O u ω £α. u n o «Γ m /*\ Li Z i-1 ω H cn

o o in

Z Ū Ό m £ 4810 oo

SCHEME

ini o] ¢)1 «' -o ^ Ή o 10 o δ Ot o o M u (0 SZ 2 ϋ r-\ &lt;0 X5

SS a u

20 α&gt; -σt « Ο '-ν Li Η -Η V ^ 2 γΗ •Η rH hJ iflfel O u

CJ&quot; 25

&gt;=o 2 —' S CD m ! (Π

* x Ο n V o £ U O , &lt;·· H J u fd -Q ip &lt;3 30 - 49 - SCHEME 9 pivaloyl chloride, ph ho2c- 69 (Aldrich)

Et 3N, THF;A-rph °^NLi 0 LV10425 o o 70

Ph

Ti(0-i-Pr)Cl3&gt; i-Pr2NEt, CN

__— O ' 3 PC02, CH3OH, ^N^^OjCHj CHC1, Ο O h,. 71

NaHC03, CHjCN

NaN(TM5) 2, DMF -15°C to 25°C BOCl/ 1 1 0

EDC, HOBT, DMF, i-Pr2NEt, 54

BOCN r&quot;&quot;r •N.

•OjH

O 75 50 SCTTEME 9 CONT'D.

BOCN Γ^^ηο iH3 'N-EC o 76

1 N NaOH, CH3OH

77 ch3

H^K^C02H o cf3co2r ck2ci2

78 o LV10425 - 51 - SCHEME 10

CH, HC1 •HjN’^-COjEC 54 isobutyl- chlorofocnate._

Echyl acocaco,

4- rtwc hylrrorpholina Cbz-Gly, -15°C CH.

CbzNH— 79

20¾ Pd(OH)j. AcOH. CHjOH CHj **

NH'^-COjEC 80

BOCN 45 £ Y°

LlNCTbSJļ THF. -78°C than allyl brorrtde 0eC

LiOOH

BOCN 82 EDC, HOBT, DMF 1-PrjNEC 80

52 52 5 10

SCHEME 10 CONT'D. 83

Os 04, NālO^

JDH O CHj O Θ4

1 N NaOH. CHjOH 15

, OH /—\ h«ž—( o 9¾ ^NvJ^°Vn-Anh^o2h o 85 20

Et3SiH. CFjCOjH. CHjCIj 25 8δ 30 - 53 - SCHEME.il LV10425

DMF. NaNCTM3)3 allyl brorride, 0°C

Θ8 1. n-BuLl. THF, 2. 10, -23° for -78°C 2 hra

aceCone Jones reaģent

BOCN

// Nn

COjH 91

EDC, HDBT, Νζ i-Pr) jEt, DMF

BOCN // CHj 1 &quot; HCl •H,N'^^C03Et 54

ςΗ, S NH^N-^OjEC

1N NaOH 92 54 SCHEME 11 CONT'D.

TFA/CH2Cl2

- 55 -LV10425 SCHEME 12

EtOAc

56 BOCN

PhaP· la inridazols 100 SCHEME 13

BOCN I 101

BOCN OCHj CHļO^ Cf 1_03

Ph3P CHjCN. 80°

BOCN Ολ 1 04

NaN(TTE)2, THF

PPh3I 1 02

Hz 1 0% Pd/C EtOAc

O

NaN(Ti«)2 BrCHjCOjEC, DMF. 0°

BOCN

NH \^C02EC

O 1 07 - 57 -5

BOCN 10 15 20 25

SCHEME 13 CONT·D 107

1 N NaOH CHjOH

N^COjH

O 108

EDC, HDBT. DMF. ΕΠΝζΙ-Ρτ), CH, I HCINHj'N^-COjEC 57. Et 109 1 N NaOH CHjOH

BOCN

HN (f^l Ο ψ^ι ^&gt;Yn'-Anh'^co3h 110 CF,COjH CHjCl,

LV10425 30 111 58 SCHEME 14

iH

HN

benzylchlorof ornate, THF, NCi-Pr)jEt, -15°C

Cbz-N 1 1 2 PPh3/irridazole/I2

Cbz-N Y ^

113 O

NaNC3&gt;B)2i DMF, HN^)

O

Cbz-N Y ^ \_/ 114 LDA. THF. -78°C Chen allyl bromide

O

Cbz-N 1 1 5 CC1*. CHjCN, HzO RuCIj, NaI04

O

COjH

Cbz-N 1 1 6 - 59 - LV10425 SCHEME 14 CONT'D.

02H O 1J_6 isobutyl chloroforrrate, NEt3, dioxa then H20, Na2C03,

O

II h2n /\^·ρ(οη)2 o

1 1 7 CH3OH, HOAc, Pd(OH)2

60 SCHEME 15 h2nH^&gt;-co2h 1 19 see, Chem Ber. 76, 1019 (1943) h2n~\ &gt;-co2h 120

a) SCC12, CH3OH

b) benzyl chlorof orrrat e, DMF, N(l-Pr)2EC

CbzHN—( V-C02CH3 1 21 LDA, THF, -78°C chen allyl brorride

CbzHN'

H2N co2ch3 1 22

crifluoroacecic acld, anisole, CH2C12, O0 C co2ch3 123 - 61 -LV10425 SC5EME 15 CONT'0

1 23

toluene, 170°C

DMF, NaN( TMS) 2 trhen 1_0

•BOCN

RuC13, NaIOv CH3CN/H20/CC14 1 26 62

SCHEME 15 CONT'D

62 BOCN

EDC, HOBT, DMF, N(i-Pr)2Et, H2N'^^02t Bu

trifluoroacetic acid, anisole, CH2C12

HN

- 63 - LV10425

Preparation of 3-(Indol-3-yl)propanol-tert-butyl- dimethvlsilvl e.ther (2L)_

To a stirring solution of 3-indolepropanol 1 (15 g, 86 mmol), DMF (200 mL), and imidazole (12.8 g, 0.19 mol) at 0eC was added tert-butyldimethylsilyl chloride (14.2 g, 95 mmol) followed by removal of the cooling bath. After 20 hours the reaction mixture was diluted with ether and then washed with H20 (2 times) and brine, dried (MgSC^), and concentrated to yield the silyl ether Z (29 g) as an amber oil. TLC Rf = 0.54 (207. ethyl acetate/hexanes); 1H NMR (CDCI3) δ 8.07 (bs, 1Ξ), 7.77 (d, J=7Hz, 1H), 7.49 (d, J=7Hz, -1H), 7.33 (t, J=7Hz, 1H), 7.26 (t, J=7Hz, 1H), 7.12 (s, 1H), 3.84 (t,_J=6Hz, 2H), 2.95 (t, J=7Hz, 2H), 2.08 (m, 2H), 1.08~(s, 9H), 0.25 (s, 3H), 0.22 (s, 3H).

Preparation of N-Acetyl-3-(indol-3-yl)propanol-tert- butvldimethvlsilvl ether (3)_ A solution of the indole 2 (29 g, 86 mmol), CH2C12 (450 mL), 1,8-diazobicyclo[5.4.0]undec-7-ene (38 mL, 0.26 mol), 4-dimethylaminopyridine (1.0 g, 8.5 mmol), and acetic anhydride (32 mL, 0.34 mol) was stirred for 1 week at ambient temperature. The reaction mixture was concentrated and then diluted with ether. The ether was then washed with H20, 57. KHSO4 and brine, dried (MgSŪ4), and concentrated. Flash chromatography (silica, 57. ethyl acetate/hexanes) gavē the acylated product 2. (27 g) as a yellow oil. TLC Rf= 0.56 (207. ethyl acetate/hexanes). 64

Preparation of N-Acetvl-3-(indol-3-vl)propanol (4)

To a stirred solution of the silyl ether 2 (27 g, 81 mmol) in THF (270 mL) at ambient temperature was added a premixed solution of n-Bu^NF (1M in THF: 244 mL, 0.24 mol) and AcOH (14 mL, 0.24 mmol) (1:1). After 20 hours the reaction mixture was diluted with ether and then vashed with H20 (2 times) and brine, dried (MgSO^), and concentrated to give the alcohol 4 (19 g) as a yellow crystalline solid. TLC Rf= 0.35 (607. ethyl acetate/hexanes); XH NMR (CDC13) δ 8.42 (m, 1H), 7.55 (d, J=7Hz, 1H), 7.36 (t, J=7Hz, 1H), 7.29 (t, J=7Hz, 1H), 7.27 (7d, J=7Hz, 1H), 7.22 (s, 1H), 3.76 (t, J=7Hž, 2H), 2.82 (t, J=7Hz, 2H) 2.61 (s, 3H), 2.00 (m, 2H).

Preparation of 5-(N-Acetyl-indol-3-yl)pent-2-enoic acid ethvl ester (5)__-

To a stirring solution of oxalyl chloride (11.4 mL, 0.13 mol) in CH2C12 (440 mL) at -78eC was added dry DMS0 (12.4 mL, 0.17 mol) dropwise. After 5 minūtes, gas.evolution ceased and the alcohol 4 (19 g, 87 mmol) in CH2C12 (40 mL) was added. After 30 minūtes, NEt2 (73 mL, 0.52 mol) was added to effect a thick slurry. The cooling bath was removed and the reaction stirred for an additional 15 minūtes before adding (carbethoxymethylene)triphenyl phosphorane (33.5 g, 96 mmol). After 2.0 hours, the reaction mixture was diluted with ether and then washed with H20 (2 times), 57. KHSO4 and brine, dried (MgSO^), and concentrated. Flash chromatography (207. ethyl acetate/hexanes) gavē the olefin 5. (14 g) as a white solid. - 65 - LV10425 TLC Rf= 0.54 (60¾ ethyl acetate/hexanes); lE NMR (CDCI3) 8.42 &lt;bd, 1H), 7.50 (d, J=7Hz, 1H), 7.34 (t, J=7Hz, 1H), 7.28 (t, J=7Hz, 1H), 7.19 (bs, 1H), 7.03 (dt, J-18 and 7Hz, 1H), 5.88 (d, J=18Hz, 1H), 4.19 (q, J=7Hz, 2H), 2.87 (t, J=7Hz, 2H), 2.63 (m, 2H), 2.61 (s, 3H), 1.28 (t, J=7Hz, 3H).

Preparation of N-(R)-a-Methylbenzyl-3(R)-[2-(indol-3-yl)ethyl]0-alanine ethyl ester (6a) and N-(R)-a-Methylbenzyl-3(S)-[2-(indol-3-yl)ethyl]0-alanine ethvl ester (6b)___ A mixture of olefin 1 (2.77 g, 9.7 nunol) and R-( + )-a-methylbenzylamine (5.03 mL, 39 nunol) was heated under a cold finger at 110eC for 40 hours._ The cooled reaction mixture was applied directly to a flash chromatography column (silica, 40:2:1, hexanes:ethyl acetate: 2-propanol). The (R,R) isomer 6a eluted first (1.19 g) as a viscous yellow oil which solidified on standing. Recrystallization from hexanes/ethyl acetate provided crystalline material.. The (R.S) isomer eluted next (1.55 g) as a viscous yellow oil containing ca 107· of the (R,R) isomer. 6a: Rf=0.52 (607· EtOAc/hexanes); *Η NMR (400 MHz, CDCI3) 5 7.84 (br s, 1H), 7.52 (dd, J=7.9, 0.7 Hz, 1H), 7.20-7.35 (m, 6H), 7.16 (tm, J=7.1, 1.3 Hz, 1H), 7.08 (tm, J=7.3, 1.1 Hz, 1H), 6.70 (br d, J=2.4 Hz, 1H), 4.10 (q, J=7.1 Hz, 2H), 3.90 (q, J=6.6 Hz, 1H), 2.80-2.90 (m, 2H), 2.68 (ΑΒΧ dt, J=16, 7.9 Hz, 1H), 2.53 (ΑΒΧ dd, J=14.5, 5.9 Hz, 1H), 2.42 (ΑΒΧ dd, J=14.6, 5.3 Hz, 1H), 1.79 (q, J=7.5 Hz, 2H), 1.33 (d, J=6.4 Hz, 3H), 1.22 (t, J=7.1 Hz, 3H). 66

Rf=0.42 (607. EtOAc/hexanes); ΧΗ NMR &lt;400 MHz, CDCI3) δ 7.95 . (br s, 1H), 7.57 (dd, J=7.5, 0.7 Hz, 1H), 7.34 (dm, J= 8.1, 0.7 Hz, 1H), 7.17-7.30 (m), 7.11 (tm, J=7.9, 0.9 Hz, 1H), 6.89 (br d, J=2.2 Hz, 1H), 4.02-4.15 (ΑΒΧ m, 2H), 3.89 (q, J=6.6 Hz, 1H), 2.95 &lt;m, 1H), 2.82 (ΑΒΧ ddd, J=15, 9.7, 5.9 Hz, 1H), 2.69 (ΑΒΧ ddd, J=15, 9.7, 6.0 Hz, 1H), 2.47 (ΑΒΧ dd, J=15.0, 5.1 Hz, 1H), 2.40 (ΑΒΧ dd, J=15.0, 7.7 Hz, 1H), 1.96 &lt;m, 1H), 1.83 (m, 1H), 1.30 (d, J=6.6 Hz, 3H), 1.21 (td, J=7.1, 0.7 Hz, 3H).

Preparation of 3(R)-[2-(indol-3-yl)ethyl]B-alanine ethvl ester (7a)____

Amine &amp;a (996 mg, 2.74 mmol) was dissolved in 10 mL EtOH. After addition of Pearlman’s catalyst (207. Pd(0H&gt;2/C, 128 mg) the flask was charged with hydrogen and maintained at balloon pressure. After 16 hours an additional portion of catalyst was added (122 mg) along with fresh H£. Four hours later the sample was filtered through celite and concentrated to provide amine Za (707 mg, 99%, ca 957. pure).:

Rf=0.22 (10:1, NH3 satd. CHCI3:EtOAc); ^ NMR (400 MHz, CDCI3) δ 8.01 (br s, 1H), 7.60 &lt;dt, J=8.9, 0.4 Hz, 1H), 7.35 (dt, J=8.1, 0.9 Hz, 1H), 7.19 (td, J=7.1, 1.3 Hz, 1Ξ), 7.11 (td, J=7.1, 1.2 Hz, 1H), 6.99 (br d, J=2.2 Hz, 1H), 4.15 (q, J=7.1 Hz, 2H), 3.72 (q, J=7.0 Hz, 1H), 3.29 &lt;m, 1H), 2.92-2.78 (m, 2H), 2.53 (ΑΒΧ dd, J=15.6, 4.0 Hz, 1H), 2.33 (ΑΒΧ dd, J=15.6, 8.8 Hz, 1H), 1.92-1.73 (m, 2H)&gt; 1.25 (q, J=7.1 Hz, 3H). - 67 - LV10425

Prepacation of N-BOC-4-pjperidineethanol (9)^

To a stirred solution of 4-piperidineethanol £ (18.7 g, 0.14 mol) and DMF (200mL) at 0°.C was added N-tert-butoxycarbonyl anhydride (31 g, 0.14 mol). After 1 hr the cooling bath was removed and the reaction mixture stirred for 20 hr. The reaction mixture was diluted with ether and then washed with water (2x) and brine, dried (MgSO^.), and concentrated to furnish £ (26 g, 62%) as a colorless oil. TLC Rf = 0.25 (40% ethyl acetate/hexanes); ΧΗ NMR (300 MHz, CDCI3) 6 4.09 (bs, 2H), 3.72 (t, J= 7 Hz, 2H), 2.70 (m, 2H), 1.75-1.10 (m, 7H), 1.46 (s, 9H) .

Preparation of N-B0C-4-piperidine ethvl iodide (10).

To a stirring solution of £ (18.0 g, 77 minol), triphenylphosphine (22.2 g, 85 mmol), imidazole (7.9 g, 115 mmol), and benzene (800 mL) af ambient temperature was added iodine (22.0 g, 85 mmol). After 5 min the heterogeneous reaction mixture was filtered and the filtrate concentrated. Flash chromatography (silica gel, 10% ethyl acetate/hexanes) gavē 1£ (20 g, 597.) as an oil. TLC Rf = 0.95 (50% ethyl acetate/hexanes); !h NMR (300 MHz, CDCI3) δ 4.11 (m, 2H), 3.24 (t, J= 6 Hz, 2H), 2.72 (m, 2H), 1.82 (dt, J = 7, 7 Hz, 2H), 1.75, -1.55 (m, 5H), 1.48 (s, 9H), 1.12 (m, 2H). 68

Preparation of N-BOC-4-piperidine ethvl azide._(ll). A soļution of Ιϋ (5.0 g, 14.7 mmol), DMSO (75 mL), and NaN3 (1.9 g, 29.4 mmol) was heated at 70® C for 2 hr. The cooled reaction mixture was diluted with ethyl acetate and then washed with water (2x) and brine, dried (MgSO^.), and concentrated to afford ϋ (3.6 g, 96%) as a colorless oil. TLC Rf = 0.75 (30% ethyl acetate/hexanes); 1H NMR (300 MHz, CDCI3) δ 4.11 (m, 2H), 3.36 (t, J= 7 Hz, 2H), 2.73 (m, 2H), 1.70 (m, 3H), 1.49 (s, 9H), 1.15 (m, 2H).

Preparation of N-BOC-4 piperidine ethyl amine (12). A mixture of H (1.1 g, 4.3 mmol), 10 7. Pd/C (0.16g), and ethanol was stirred under a hydrogen atmosphere (1 atm) for 1.5 hr. The reaction mixture was then filtered through a celite pad and the filtrate concentrated to give crude 12 (1.0 g) as an oil. TLC Rf = 0.18 (9:1:1 CH2CH2/CH30H/H0Ac); 1H NMR (300 MHz, CDCI3) δ 4.10 (m, 2H), 2.78 (t, J= 7 Hz, 2H), 2.70 (m, 2H), 1.80 (m, 2H), 1.67 (m, 2H), 1.52 (m, 1Ξ), 1.47 (s,9H), 1.17 (m, 2H).

Preparation of l-[2-(N~Boc-piperidin-4-yl)ethyl]-3- propen-2-vl-(2-imidazolidinone) (14)_

To a stirred soļution of 12 (1.5 g, 17 mmol) in DMF (75 mL) at ambient temperature was added LiN-(TMS)2 (1M in hexanes, 17 mL) to effect a precipi-tate. Allyl bromide (1.6 mL, 19 mmol) was then added to the reaction mixture. After 15 min the - 69 - LV10425 homogeneous mixture was treated again with LiN(TMS)2 (14 mL) followed by the iodide (5.9 g, 17 mmol) after 5 min. The reaction was stirred for 20 hr then diluted with ether. The ether portion was washed with water (2x), 5% KHSO^ and brine, dried (Mg SO4) , and concentrated. Flash chromatography (40% ethyl acetate/hexanes) gavē 14 (0.8 g, 15%) as a colorless oil. TLC Rf = 0.46 (70% ethyl acetate/hexanes);. NMR (300 MHz, CDCI3) 5 5.80 (m, 1H), 5.23 (m, 2H), 4.10 (m, 2H), 3.82 (m, 2H), 3.30 (m, 6H), 2.72 (m, 2H), 1.70 (m, 3H), 1.50 (m, 2H), 1.46 (s, 9H), 1.15 (m, 2H).

Preparation of l-[2-(N-Boc-piperidine-4-yl)ethyl])- 3-acetic acid-(2-imidazQlidinone_) . (15) .—--

To a vigorously stirred solution of 14 (450 mg, 1.4 mmol), CCI4, acetonitrile, and water at. ambient temperature was added RUCI3 (12 mg, 4.4 mol%) and excess NalO^.. After 60 hr the reaction was filtered through a celite pad washing with ethyl acetate. The filtrate was extracted with sat NaHC03 followed by acidifying the aqueous phase to pH 3 with 5% KHSO4. The acidic aqueous phase was then extracted with ethyl acetate (2x) and the organic portion was dried (Mg SO4) and concentrated. Flash chromatography (silica gel, 9:0.2:0.2 CH2C12/CH30H/H0Ac) gavē JJ. (60 mg, 12%) as a colorless oil. TLC Rf = 0.29 (9.5:0.5:0.5 CH2C12/CH30H/H0Ac); 1H NMR (300 MHz, CDC13) δ 4.05 (m, 2H), 3.90 (m, 2H), 3.45 (m, 2H), 3.40 (m, 2H), 3.24 (m,2H), 2.69 (m, 2H), 1.72 (m, 2H), 1.46 (m, 3H), 1.46 (s,9H), 1.15 (m. 2H). 70

Preparation of [l-[2-(N-Boc-piperidin-4-yl)ethyl]-(2-imidazolidinone)-3]-acetyl-3(R)-[2-(indol-3-yl)-ethyl] β-alanine ethvl ester (16)__ A stirred mixture of 15. (60 mg, 0.17 mmol), 7a (66 mg, 0.25 mmol), HOBT (30 mg, 0.22 mmol), triethylamine (31 uL, 0.22 mmol), and DMF (1.1 mL) at 0eC was treated with EDC (42 mg, 0.22 mmol) folloved by removal of the cooling bath. After 20 hr the reaction was diluted with ethyl acetate and then washed with water, 5% KHSO4, and brine, dried (MgSO^), and concentrated. Flash chromatography (silica gel, ethyl acetate) gavē 16. (35mg, 357.) as a colorle'ss oil. TLC Rf = 0.20 (ethyl acetate); ^ NMR (300 MHz, CDCI3) 5 8.07 (bs, 1H), 7.57 &lt;d, J = 8 Hz, 1H), 7.37 (d, J = 8 Hz, 1H), 7.18 (t, J = 8 Hz, 1H), 7.11 (t, J = 8 Hz, 1H), 7.05 (bs, 1H), 6.89 (d, J = 9 Hz, 1H), 4.37 (m, 1H), 4.13 (q, J = 7 Hz, 2H), 4.07 (m, 2H), 3.83 (dd. J = 18 Hz, 2H), 3.43 - 3.20 (m, 6H), 2.80 (m, 2H), 2.56 (d, J = 5 Hz, 2H), 2.00 - 1.05 (m, 9H), 1.47 (s, 9H), 1.25 (t, J = 7 Hz, 3H).

Preparation of [l-[2-(N-Boc-piperidin-4-yl)ethyl]-(2-imidazolidinone)-3]-acetyl-3(R)-[2-(indol-3-yl)ethyl]- β-alanine (17) ----- A mixture of 16. (30 mg, 50 umol), 1N, NaOH (0.2 mL) and ethanol was stirred at arabient tempera-ture for 1 hr. The reaction mixture was then diluted with ethyl acetate and 57 KHSO4 an&lt;^ orSanic portion washed with water and brine follov/ed by drying (MgSŪ4) and concentration to furnish H (30 mg, 1007.) as a colorless oil. TLC Rf = 0.74 (9:1:1 CH2C12/CH3OH/HOAc). - 71 - LV10425

Preparation of [l-[2-piperidin-4-yl)ethyl]-(2-im idazolidinone)-3]-acetyl-3(R)-[2-(indol-3-yl)ethyl] β-alanine (18)_

To a stirring solution of 12 (30 mg, 50 umol), dichloromethane (300 uL), and anisole (15 uL, 100 umol) at - 15eC was added TFA (0.3 mL). After 20 min the reaction mixture was concentrated and the residual TFA removed azeotropically with toluene.

Flash chromatography (silica gel, 10:0.8:0.8 methanol/ NH4&gt;/water) gavē 18. (15 mg, 66%) as a white solid. TLC Rf = 0.16 (10:1:1 methanol/NH40H/water); ΧΗ NMR (300 MHz, CD3OD) δ 7.44 (d, J = Hz, 1H), 7.20 (d, J = 8 Hz, 1H), 6.95 (t, = J = 8 Hz, 1E), 6.94 (s, 1H), 6.88 (t, J = 8 Hz, 1H), 4.17 (m, 1H), 3.71 (s, 2H), 3.40)3.10 (m, 8H), 2.86 (m, 2H), 2.70 (t, J = 6 Hz, 2H), 2.32 (m, 2H), 1.90 (m, 4H), 1.55 (m, 1H), 1.43 (m, 2H), 1.28 (m, 2H).

Preparation of l-[2-(N-Boc-piperidin-4-yl)ethyl]-(2- pvrrolidinone) (19)-

To a stirred solution of 12 (2.7 g, 11.8 mmol) acetonitrile (60 mL), and diisopropyl ethylamine (4.1 mL 23,6 mmol) at 0®C was added 4-chlorobutyryl chloride (2.6 mL, 23.6 mmol) followed by removal of the cooling bath. After 5 hr the reaction mixture was diluted with ethyl acetate and then washed with water (2x) and brine, dried (MgSŪ4), and concentrated. The crude amide was dissolved in DMF (60 mL) cooled to 0°C then treated with NaN(TMS)2 (1M in THF, 11.8 mL). After 5 min the reaction mixture was diluted with ethyl acetate and then 72 washed with water and brine, dried (MgS04), and concentrated. Flash chromatography (silica gel, 70% ethyl acetate/hexanes); gavē 12. (0.4 g, 12%) as a colorless oil. TLC Rf = 0.27 (70% ethyl acetate/hexanes); 1H NMR (300 MHz, CDCI3) δ 4.10 (m, 2H), 3.39 (t, J = 7 Hz, 2H), 3.34 (t, J= 7 Hz, 2H), 2.70 (m, 2H), 2.40 (m, 2H), 2.05 (m, 2H), 1.72 (m, 2H), 1.46 (m, 3H), 1.45 (s, 9H), 1.13 (m, 2H).

Preparation of l-[2-(N-Boc-piperidin-4-yl)ethyl]-3- propen-2vl-(2-Pvrrolidinone) (20)_

To a stirred solution of 12(325 mg, 1.1 mmol) in THF ( 5 mL) at -78eC was added LDA (0.5 M in THF 2.4 mL) dropvise. After 15 min allyl bromide (0.16 mL, 2.2 mmol) was added and the reaction stirred at -78eC for 1 hr followed by guenching with HOAc (0.1 mL). The reaction mixture was then diluted with ethyl acetate and then vashed with water and brine, dried (MgS04), and concentrated. Flash chro.m atography (silica gel, 507. ethyl acetate/hexanes) gavē 22 (160 mg, 45%) as an oil. TLC Rf-0.23 (507. ethyl acetate/hexanes) ; ΧΗ NMR (300 MHz, CDCI3) 5 5.80 (m, 1H), 5.10 (m, 2H), 4.10 (m, 2H), 3.32 (m, 4H), 2.70 (m, 2H), 2.56 (m, 2H), 2.20 (m, 2H), 1.73 (m, 3H), 1.48 (s, 9H), 1.13 (m, 2H). - 73 - LV10425

Preparation of l-[2-(N-Boc-piperidin-4-yl)ethyl]-3- asetic acid-_2-pvrrolidinone (21)_____

Utilizing the procedure for converting 14 to 12, 20. (130 mg, 0.4 mmol) gavē 21 (80 mg, 48¾) as an oil after flash chromatography (silica gel, 9:0.2:0.2 CH2C12/CH30H/H0Ac). TLC Rf=0.25 (9:0.2:0.2 CH2C12/CH30H/H0Ac); 1H NMR (300 MHz, CDC13) δ 4.05 (m, 2H), 3.40 (m, 4H), 2.96 (m, 1H), 2.83 (m, 1H), 2.70 (m, 2H), 2.55 (m, 1H), 2.40 (m, 1H), 1.90-1.10 (m, 7H), 1.48 (s, 9H).

Preparation of [l-[2-(N-Boc-piperidin-4-yl)ethyl]-2-pyrrolidinone-3]acetyl-[2-(indol-3-yl)-ethyl]P- alanine ethvl ester (22)_:_

Utilizing the procedure for converting 15. to 16. 21 (120 mg, 0.35 mmol) gavē 22. (90 mg, 48%) after flash chromatography (silica gel, ethyl acetate). TLC Rf=0.40 (ethyl acetate); λΕ NMR (400 MHz, CDC13) δ 8.09 (bd, 1H), 7.58 (d, J=8 Hz, 1H), 7.36 (d, J=8 Hz, 1H), 7.17 (t, J=8 Hz, 1H), 7.10 (t, J=8 Hz, 1H), 7.07 (bs, 1H), 4.39 .(m, 1H), 4.12 (q, J=7 Hz, 2H), 4.05 (m, 2H), 3.30 (m, 4H), 2.90-2.25 (m, 8H), 2.00-1.00 (m, 8H), 1.48 (s, 9H), I. 23 (t, J=7 Hz, 3H).

Preparation of [l-[2-(N-Boc-piperidin-4-yl)ethyl]-2-pyrrolidinone-3]acetyl-[2-(indol-3-yl)-ethyl]p- alanine (23)-------

Utilizing the procedure for converting 1£ to II, 22 (80 mg, 0.14 mmol) gavē 22 (80 mg) after work-up, which was used directly for the next reaction. TLC Rf=0.50 (9:1:1 CH2Cl2/CH30H/H0Ac). 74

Preparation of [l-[2-(piperidin-4-yl)ethyl]-2-pyrroli dinone-31acetvl-r2-(indol-3-vl)-ethvnB-alanine-(£4J— Utilizing the procedure for converting IZ to 18, 21 (80 mg, 0.14 mmol) gavē 24 (15 mg, 237.) after flash chromatography (silica gel, 10:0.65:0.65 methanol/NH40H/water). TLC Rf=0.53 (10:1:1 methanol/NH4OH/water); ^ NMR (300 MHz, CD3OD) δ 7.47 (t, J=7 Hz, 1H), 7.25 (d, J=7 Hz, 1H), 6.99 (m, 2H), 6.88 (t, J=7 Hz, 1H), 4.19 (m, 1H), 3.49 (m, 0.5 H), 3.40 (m, 0.5H), 3.34-2.95 (m, 6H), 2.90-2.60 (m, 4H), 2.48 (m, 1H), 2.40-2.20 (m, 4H), 2.13 (m, 1H), 2.00-1.05 (m, 11H).

Preparation of .5-chloro-valerovl-amino-2-pr.Qpene (28.1 To a stirred mixture of allyl amine 21 (2.0 g, 27 mmol), NMM (8.9 mL, 81 mmol), and acetonitrile (140 mL), at ambient temperature was added 4-chloro-butyryl chloride (3.4 mL, 27 mmol). After 20 hours the reaction mixture was concentrated. The residue was dissolved in ethyl acetate and then washed with water, 57. KHS04 and brine, dried (MgS04), and concentrated to give 21 (4.1 g, 877.) as a yellow oil. TLC Rf=0.17 (40% ethyl acetate/hexanes). 1.

Preparation of l-(propen-2-vl)-2-piperidone C271-

To a stirred solution of 21 (2.0 g, 11 mmol) in THF (110 mL) 'at -78°C was added NaN(TMS)2 (1-0 M in THF, 11 mL) in a stream followed by removal of the cooling bath. After 1.5 hours the reaction mixture was diluted with ethyl acetate and then washed with water, 57. KHS04 and brine, dried (MgS04), and concentrated. Flash chromatography (silica gel, 607. - 75 - LV10425 ethyl acetate/hexanes) gavē 22 (1.1 g, 72%) as an oil. TLC Rf=0.15 (607. ethyl acetate/hexanes); lE NMR (300 MHz, CDCI3) δ 5.78 (m, 1H), 5.15, 4.00 (d, J=7 Hz, 2H), 3.24 (m, 2H), 2.41 (m, 2H), 2.07 (m, 2H), 1.80 (m, 4H).

Preparation of 3-[2-(N-Boc-piperidin-4-yl)ethyl]-l- acetic acid-2-piperidone (28)_

To a stirred solution of 22 (1.0 g, 7.2 mmol) and THF (70 mL) at -78*C was added LDA (0.5 M in THF, 15.8 mL) dropvise. After 15 minūtes iodide 10 (2.7 g, 7.9 mmol) in THF (5 mL) was added folloved by slow varming to -30eC over a 1 hour period. The reaction was quenched with acetic acid (0.2 mL) then concentrated. Flash chromatography (silica gel, 407. ethyl acetate/hexanes) gavē 23 (1.5 g, 59%) as an oil. TLC Rf=0.58 (ethyl acetate); NMR (300 MHz, CDCI3) δ 5.77 (m, 1H), 5.15 (m, 1H), 4.06 (m, 2H), 4.00 (m, 2H), 3.23 (m, 2H), 2.59 (m, 2H), 2.19 (m, 1H), 2.05-1.25 (m, 11H), 1.48 (s, 9H)., 1.10 (m, 2H).

Preparation of 3-[2-(N-Boc-piperidin-4-yl)ethyl]-3- acetic acid-2-piperidone (29)_ A vigorously stirred mixture of 28 (0.84 g, 2.4 mmol), CCI4 (5 mL), acetonitrile (5 mL), NaI04 (2.1 g, 9.8 mmol), and water (7.5 mL) at ambient temperature was treated with RUCI3 (50 mg, 0.24 mmol). After 20 hours additional RUCI3 ((50 mg) was added. After 4 hours the reaction mixture was diluted with ethyl acetate and half sat NaHC03·

After shaking and separation the aqueous portion was 76 acidified with 57. KHSO4, and extracted with ethyl acetate. The organic extracts were washed with brine, dried (MgS04&gt;, and concentrated. Flash chromatography (silica gel, 9:0.2:0.2 CH2Cl2/methanol/ HOAc) gavē 21 (280 mg, 307·) as an oil. TLC Rf=0.36 (9:0.5:0.5 CH2Cl2/methanol/HOAc).

Preparation of [3-[2-(N-Boc-piperidin-4-yl)ethyl]-2-imidazolidinone-l]acetyl-[3(R)-[2-(indol-3-yl)ethyl]- β-alanine ethvl ester (30)__

Utilizing the procedure for converting H to 16. 21 (280 mg, 0.76 mmol) gavē (300 mg, 64¾) as an oil after flash chromatography (silica gel, 857. ethyl ačetate/hexanes). TLC Rf=0.18 (857. ethyl acetate/hexanes); 1H NMR (300 MHz, CDCI3) 5 8.14 (m, 1H), 7.60 (d, J=8 Hz, 1H), 7.39 (d, J=8 Hz, IR), 7.20 (t, J=8 Rz, 1H), 7.13 (t, J=8 Hz, 1H), 7.05 (bs, 1H), 6.90 (d, J=9 Hz, 1H), 4.35 (m, 1H), 4.11 (q, J=7 Hz, 2H), 4.05 (m, 2H), 3.98 (s, 2H), 3.40 (m, 2H), 2.90-2.50 (m, 6H),. 2.33 (m, 1H), 2.00-1.00 (m, 15H), 1.48 (s, 9H), 1.27 9t, J=7 Hz, 3H). 30 - 11 - LV10425

Preparation of [3-[2-(N-Boc-piperidin-4-yl)ethyl]-2-piperidone-l]acetyl-[3(R)-[2-indol-3-yl)ethyl]P- alanine (31)____

Utilizing the procednre for converting H to IZ, 33 (160 mg, 0.26 mmol) gavē 31 (140 mg, 92%) as a white solid after work-up. TLC Rf=0.59 (9:1:1 CH2Cl2/methanol/H0Ac).

Preparation of [3-[2-(piperidin-4-yl)ethyl]-2-piper-idone-l]acetyl-[3(R)-[2-indol-3-yl)ethyl]P- a ļaujas ..13.2)_—

Utlizing the procedure for converting H to 18. 31 (140 mg, 0.24 mmol) gavē 32. (55 mg, 47%) as a white solid after flash chromatography (silica gel, 10:0.4:0.4 methanol/NH^OH/vater). TLC Rf=0.46 (10:1:1 methanol/NH^OH/ttater); XH NMR (300 MHz, CD3OD) δ 7.54 (d, J=8 Hz, 1H), 7.30 (d, J=8 Hz, 1H), 7.03 (t, J=8 Hz, 1H), 7.02 (bs, 1H), 6.97 (m, 1H), 4.28 (m, 1H), 4.20 (d, J=16 Hz, 0.5H), 4.12 (d, J=16 Hz, 0.5H), 3.80 (d, J=16 Hz, 0.5H), . 3.75 (d, J=1.6 Hz, 0.5H), 3.50-3.15 (m, 4H), 2.80 (m, 4h), 2.50-2.30 (m, 3H), 2.00-1.20 (m, 15H).

Preparation of 3-[2-(N-Boc-piperidin-4-yl)ethyl]-2- py r r.o Li dipos.g (36a)_—

To the benzyl amide 35a (0.8 g, 2.2 mmol) in THF (20 mL) at -78eC was added freshly prepared lithium di-tert-butylbiphenyl (0.5 M in THF, 10.8 mL) 78 in 2 portions. After 1 hour the reaction mixture was quenched with HOAc (0.1 mL) followed by concentration. Flash chromatography (5% methanol/ ethyl acetate) gavē 36a (470 mg, 72%) as a white solid. TLC Rf=0.27 (10% methanol/ethyl acetate); ΧΗ NMR (300 MHz, CDCI3) δ 6.03 (m, 1H), 4.10 (m, 2H), 3.35 (m, 2H), 2.70 (m, 2H), 2.33 (m, 2H), 1.92 (m, 1H), 1.80 (m, 1H), 1.70 (m, 2H), 1.50-1.30 (m, 4H), 1.48 (s, 9H), 1.12 (m, 2H).

Preparation of Methyl l-acetate-3-[2-(N-Boc-piperidin- 4-vl)ethvn-2-pYrrolidinone (37a)_.

To_a stirred solution of 36a (340 mg, 1.1 minol) in THF (10 mL) at -78eC was added NaN(TMS)2 (1.0 M in THF, 1.1 mL). After 15 minūtes ethyl bromoacetate (250 uL, 2.2 mmol) was added and the reaction mixture to warm to -20°C. After 2 hours the reaction was quenched with HOAc (0.1 mL) and then concentrated. Flash chromatography (silica gel, 50% ethyl acetate/hexanes) gavē 37a (380 mg, 907.) as an oil. TLC Rf=0.66 (10% methanol/ethyl acetate); *H NMR (300 MHz, CDCI3) δ 4.22 (q, J=7 Hz, 2H), 4.10 (m, 2H), 4.08 (s, 2H), 3.24 (m, 2H), 2.70 (m, 2H), 2.44 (m, 1H), 2.26 (m, 1H), 2.00-1.65 (m, 4H), 1.48 (s, 9H), 1.50-1.30 (m, 4H), 1.31 (t, J=7 Hz, 3H), 1.12 (m, 2H). - 79 - LV10425

Preparation of 3-[2-(N-Boc-piperidin-4-yl)ethyl]-2- pvrrolidinone-l-acetic acid (38¾)__

Utilizing the procedure for converting 16. to 17. 37a (390 mg, 1.1. nunol) gavē 38a (340 mg, 91%) as a white foam after work-up. TLC Rf=0.39 (9:0.5:0.5 CH2Cl2/methanol/H0Ac).

Preparation of [3-[2-(N-Boc-piperidin-4-yl)ethyl]-2-pyrrolidinone-l]acetyl-[3(R)-(indol-3-yl)-ethyl]P- alanine ethvl ester (39a)__—

Utilizing the procedure for converting 12 to 16, 22a (310 mg, 0.92 mmol) gavē 12a (460 mg, 86%) as a yellow oil after flash chromatography (silica gel, 857. ethyl acetate/hexanes). TLC Rf=0.16 (85% ethyl acetate/hexanes); NMR (300 MHz, CDCI3) δ 8.21 (bs, 1H), 7.58 (d, J=8 Hz, 1H), 7.38 (d, J=8 Hz, 1H), 7.20 (t, J=8 Hz, 1H), 7.13 (t, J=8 Hz, 1H), 7.05 (bs, 1H), 6.72 (d, J-10 Hz, 1H), 4.35 (m, 1H), 4.13 (q, J=7 Hz, 2H), 4.09 (m, 2H), 3.97 (d, J=15 Hz, 1H), 3.86 (d, J=15 Hz, 1H), 3.41 (m, 2H), 2.80 (m, 2H), 2.65 (m, 2H), 2.55 (dd, J=1.5 Hz, 2H), 2.45 (m, 1H), 2.24 (m, 1H), 2.00-1.20 (m, 10 H), 1.48 (s, 9H), 1.26 (t, J=7 Hz, 3H), 1.04 (m, 2H).

Preparation of [3-[2-(N-Boc-piperidin-4-yl)ethyl-l]- acetvl-r3(,R&gt;-r2-(indol-3-vl)ethvl1p-alanine (4Qa)-

Utilizing the procedure for converting 16 to IZ, 39a (460 mg, 0.79 mmol) gavē 40a (360 mg, 86%) as a foam after work-up. TLC Rf=0.57 (9:1:1 CH2Cl2/methanol/HOAc). 80 ι «

Preparation of [3-[2-(piperidin-4-yl)ethyl-l]acetyl- r3aO-r2-(indpl-3-vl)ethvnp-alanine (4laJ-

Utilizing the procedure for converting 12 to 18. 40a (360 mg, 0.65 mmol) gavē 41a (180 mg, 597«) as a white solid after flash chromatography (silica gel, 10:0.4:0.4 methanol/NH40H/water). TLC Rf=0.21 (10:0.4:0.4 methanol/NH40H/water); XH NMR (300 MHz, CD3OD) δ 7.36 (d, J=8 Hz, 1H), 7.13 (d, J=8 Hz, 1H), 6.87 (t, J=8Hz, 1H), 6.86 (s, 1H), 6.78 (t, J=8 Hz, 1H), 4.09 (m, 1H), 3.82 (d, J=16 Hz, 1H), 3.70 (d, J=16 Hz, 1H), 3.33 (m, 2H), 3.07 (ra, 2H), 2.63 (m, 4H), 2.30 (m, 1H), 2.25 (m, 2H), 2.05 (m, 1H), 1.90-1.50 (m, 6H), 1.40-1.05 (ιηΤδΗ).

Preparation of 3-[2-(N-Boc-piperidin-4-yl)ethyl]-2- pvrrolidinone 23.61?)_.— --

Utilizing the procedure for converting 35a to 36a. 35b (0.7 g, 1.7 mmol) gavē 36b (260 mg, 627.) as a white solid after flash chromatography (silica gel, 5% methanol/ethyl acetate). TLC = Rf = 0.27 (10% methanol/ethyl acetate).

Preparation of Ethyl 3-[2-N-Boc-piperidin-4-yl)ethyl]- 2-pvrrolidinone-l-acetate (37b)___

Utilizing the procedure for converting 36a to 37a. 36b (260 mg, 0.88 mmol) gavē 37b (300 mg, 897o) as an oil after flash chromatography (silica gel, 507. ethyl acetate/hexanes) . TLC Rf = 0.66 (107o methanol/ethyl acetate). - 81 - LV10425

Preparation of 3-[2-(N-Boc-piperidin-4-yl)ethyl]-2- pvrrolidinone-l-acetic acid (38b)_

Utillizing the procedure for converting lb to II, 37b (300 mg, 0.78 mmol) gavē 38b (260 mg, 987.) as a crystalline solid. TLC Rf = 0.38 (9:0.5:0.5 CH2Cl2/methanol/HOAc).

Preparation of [3-[2-(N-Boc-piperidin-4-yl)ethyl]-2-pyrrolidinone-l]acetyl-[3(R)-[2-(indol-3-yl)-ethyl]P- alanine ethvl ester (39b)_

Utilizing the procedure for converting 12 to 12, Mh (260 mg, 0.73 mmol) gavē 12b (360 mg, 837.) as an oil after flash chromatography (silica gel, 85% ethyl acetate/hexanes). TLC Rf = 0.16 (857. methanol/ethyl acetate); ΧΗ NMR (300 MHz, CDCI3) 5 8.29 (bs, 1H), 7.57 (d, J = 8 Hz, 1H), 7.38 (d, J = 8 Hz, 1H), 7.19 (t, J = 8 Hz, 1H), 7.12 (t, J = 8 Hz, 1H), 7.04 (d, J = 1 Hz, 1H), 6.67 (d, J = 10 Hz, 1H), 4.33 (m, 1H), 4.13 (q, J = 7

Hz, 2H), 4.07 (m, 2H), 3.99 (d, J = 16 Hz, 1H), 3.82 (d, J =16 Hz, 1H), 3.40 (m, 2H), 2.80 (m, 2H), 2.68 (m, 3H), 2.55 (dd, J = 5,2 Hz, 2H), 2.43 (m.lH), 2.22 (m, 1H), 2.00 - 1.00 (m, 11H), 1.47 (s, 9H), 1.28 (t, J=7Hz, 3H).

Preparation of [3-[2-(N-Boc-piperidin-4-yl)ethyl]-2-pyrrolidinone-l]acetyl-[3(R)-[2-indol-3-yl)-ethyl^- alanine (40b).,.--

Utilizing the procedure for converting 12 to 17. 39b (280 mg, 0.47 mmol) gavē 40_h (250 mg, 94%) as a white solid after flash chromatography (silica gel, 9:1:1 CH2Cl2/methanol/H0Ac). TLC Rf = 0.21 (9:1:1 CH2Cl2/methanol/H0Ac). 82

Preparation of [3-[2-(N-piperidin-4-yl)ethyl]-2-pyrrolidinone-l]acetyl-[3 (R)-[2-(indol-3-yl)-ethyl] β-alanine (41b)_

Utilizing the procedure for converting U to 18. 40b (250 mg, 0.44 mmol) gavē 41b (100 mg, 497o) as a white solid after flash chromatography Csilica gel, 10:0.4:0.4 methanol/NH^OH/vater). TLC Rf = 0.46 (10:0.4:0.4 methanol/NH^OH/vater); J-H NMR (300 MHz, CD3OD) δ 7.52 (d, J = 8 Hz, 1H), 7.29 (d. J = 8 Hz, 1H), 7.03 (t, J = 8 Hz, 1H), 7.02 (s, 1H), 6.95 (t, J = 8 Hz, 1H), 4.27 (m, 1H), 4.05 (d, J * 16 Hz, 1H), 3.81 (d, J = 16 Hz, 1H), 3.42 (m, 2H), 3.26 (m, 2H), 2.90-2.70 (m, 4H), 2.49 (m, 1H&gt;; 2.40 (d, J_= 6 Hz, 2H), 2.22 (m, 1H), 2.05 -1.70 (m, 5H), 1.60 - 1.25 (m, 5H).

a) Swrn Oxldatlon H - b) Ph3P '^''COjEC 9

Ethvl 4-(N-B0C-piperidin-4-vl)crotonate(42)

To a stirred solution of oxalyl chloride (0.43 mL, 5.0 mmol) in CH2CI2 (20mL) at -78aC was added DMS0 (0.52ml, 7.0 mmol) dropwise. After gas evolution subsided (~5 minūtes) the alcohol 2. (0.8 g, 3.5 mmol) in CH2CI2 (20 mL) was added in a streara. After 20 minūtes triethylamine (1.7 mL, 12 mmol) was added dropwise and then the cooling bath removed. - 83 - LV10425

After 20 minūtes carbethoxymethylenetriphenyl-phosphorane (1.4g, 4.0 mmol) was added in one portion. After 2.0 hours the reaction mixture was diluted with Petroleum ether and then washed sequentially with H2O, 57. KHSŪ4, and brine, dried (MgSO^), and concentrated. Flash chromatography (silica, 15% ethyl acetate/hexanes) gavē the ester 42 as a colorless oil. TLC Rf = 0.79 &lt;50% ethyl acetate/hexanes); 1H NMR (300 MHz, CDCI3) δ 6.91 (dt,J = 16 and 7 Hz, 1H), 5.81 &lt;bd,J = 17 Hz, 1H), 4.18 (q,J = 7 Hz, 2H), 4.08 (m,2H), 2.67 (m,2H), 2.14 (t,J = 7 Hz,2H), 1.70-1.05 (m,5H), 1.44(s,9H), 1.28(t,J = 7H, 3H).

Preparation of Ethyl 4-(N-B0C-piperidin-4-yl)- butvrate (43)___

The olefin 42. (26g, 87mmol) in ethyl acetate (500 mL) was hydrogenated, at ambient temperature, under a hydrogen atmosphere (1 atm) in the presence of 107. Pd/C (5.0g) overnight. The reaction mixture was then purged with argon and filtered through a celite pad. Concentration of the filtrate folloved by flash chromatography (silica, 10% ethyl acetate/hexanes) gavē the ester 41 as a crystalline solid. TLC Rf = 0.42 (207. ethyl acetate/hexanes); *H NMR (300 MHz, CDCI3) 5 4.16 (q,J = 7 Hz,2H), 4.10 (m,2H), 2.69 (m,2H), 2.31 (t.J = 7 Hz,2H), 1.68 (m,4H), 1.38 &lt;s,9H), 1.40 (m,lH), 1.11 (m,2H). 84

Preparation of 4-(N-B0C-Piperin-4-yl)butanoic acid (44~)_ A solution of ester 42 (19g, 63ramol), ethanol (300 mL) and 1 N NaOH (100 mL, lOOmmol) was stirred at ambient temperature for 2.5 hours followed by concentration. The residue was diluted with 57. KHSO4 and ethyl acetate and then transferred to a separatory funnel. The phases were shaken then separated and then the organic portion was vashed with brine, dried (MgSC^), and concentrated to give the acid 44 as a colorless oil that crystallized upon standing. mp = 80 - 81eC; TLC Rf = 0.68 (ethyl acetate); ^ NMR (300 MHz, CDCI3) 5 4.10 (m, 2H), 2.71 (m,2H), 2.38 (t,J = 7Hz, 2H), 1.70 (m, 4H), 1.60-1.30 (m,3H), 1.48 &lt;s,9H), 1.12 (m,2H).

Preparation of 4(S)-Benzyl-2-oxazolidinone-4-(B0C-_ piperidin-4-vl) butvrate (45)__

To a stirred solution of 44 (15.3g, 56mmol), NEt3 (9.4mL, 67mmol) and dry ΤΗΓ (240mL) at -78eC was added trimethylacetyl chloride (7.6mL, 61mmol) in a stream. After 10 min. the cooling bath was removed and replaced with an icebath. After 1.0 h the heterogeneous raixture was recooled to -78eC followed by cannula addition of lithium (S)-(-)-4-benzyl-2-oxazolidinone (62mmol) in dry THF (150mL) AT -78ec with n-BuLi (38.8mL, 62mmol, 1.6 - 85 - LV10425 M/hexanes). After addition was complete, the reaction mixture was warmed to 0eC for 1.0 h, and then diluted with ethyl acetate and vashed with H2O, sat. NaHCŪ3, 5% KHSO4 and brine, dried (MgSO^.), and concentrated. Flash chromatography (silica, 30% ethyl acetate/hexanes) gavē 45. as a colorless oil. TLC Rf = 0.45 &lt;30% ethyl acetate/hexanes); 1H NMR (400 MHz, CDCI3) 7.36-7.20 &lt;m,5H), 4.67 (m.lH), 4.18 (m,2H), 4,08 &lt;m,2H), 3.30 &lt;dd,J = 13 and 3 Hz, 1H), 2.93 (m,2H), 2.77(dd,J = 13 and 10 Ηζ,ΙΗ), 2.69 (m,2H), 1.70 (m,4H), 1.50-1.30 (m,3H), 1.45 (s,9H), 1.11 (m,2H).

Preparation of 4(S)-Benzyl-2-oxazolidinone-4-(B0C-piperidin-4-vl&gt;-2&lt;R)-&lt;2-cvanoethvl)btitYratĢ_(46) —

To a stirred solution of T1CI4 (42 mL, 42mmol, 1M/CH2C12) and CH2C12 (250 mL) at 0eC was added titantium (IV) isopropoxide (4.2 mL, 14mmol). After 15 min. diisopropylethylamine (ll.OmL) in CH2C12 (75 mL) was added folloued by continued stirring at 0eC to the deep red solution. After 10 minūtes, 45 (21.8 g, 51 mmol) in CH2C12 (75 mL) was added followed by continued stirring at 0eC for 1.0 hour. Acrylonitrile (33.4 mL, 0.50 mol) was added at 0eC to the deep red solution. After 4.0 h the reaction was quenched with sat. NH^Cl (150 mL) at 0“C, then extracted with CH2C12 (3x250 mL). The combined organic extracts were vashed vith sat. NaHCŪ3 and brine, dried (MgSO^.) and concentrated. Flash chromatography (silica, 25% ethyl acetate/hexanes) - 86 gavē crude M as a yellow oil. Crude 4&amp; was chromatographed (silica, 2.57. acetone/CH2Cl2) to yield 4$. as an oil which was 977. pure by HPLC in addition to mixed fractions (1.6g). TLC Rf = 0.35 (2.57. acetone/CH2Cl2); NMR (400 MHz, CDC13) 6 7.50-7.30 (m,5H), 4.68 (m,lH), 4.21 (m,2H), 4.07 (m,2H), 3.80 (m,lH), 3.33 (dd,J = 13 and 4 Hz, 1H), 2.77 (dd, J = 13 and 10 Hz, 1H), 2.65 (m, 2H), 2.38 (m, 2H), 2.13 (m, 1H), 1.89 (m,lH), 1.75 (m, 1H), 1.63 (m,2H&gt;, 1.50 (m, 2Ξ), 1.45 (s,9H), 1.35 (m, 1H), 1.25 (m, 2H), 1.08 (m, 2H).

Preparation of 4(S)-Cyclohexylmethyl-2-oxazolidin-one-4-(B0C- piperidin-4-yl)-2(R)-(2-aminopropyl) butvrate»HCl (47)__ A mixture of M. (19.2g, 40 mmol), .Pt02 (2.0g), CH3OH (70 mL) and CHCI3 (7.0 mL) was shaken on the Parr apparatus under a hydrogen atmosphere (60 PSI) at ambient temperature for 3.0 h. The reaction mixture was filtered through a celite pad and then concentrated to furnish the crude amine«HCl 47 as a white solid. This material was used directly in the next step. TLC Rf = 0.50 (10:1:1 CH2Cl2/CH3OH/HOAc); XH NMR (400 MHz, CDCI3) δ 8.28 (bs, 2H), 4.50 (m,lH), 4.35 (m.lH), 4.14 (m.lH), 4.03 (m,2H), 3.63 (m, 1H), 3.02 (m,2H), 2.63 (m,2H), 2.00-1.00 (m,24H), 1.45 (s,9H). - 87 - LV10425

Preparation of 3-(R)-[N-B0C-2-(piperidin-4-yl)ethyl]- 2-piperidone (48)_

The crude amine«HCl 42 (16.6 g, 31 mmol), acetonitrile (750 mL), and NaHCOj (10.0 g) was stirred at ambient temperature for 20 h. The heterogeneous mixture was then treated with di-tert-butyldicarbonate (3.0g) followed by continued stirring for 1.0 h to reprotect minor amounts of free piperidine that formed in the previous reaction. The NaHCŪ3 was removed by filtration and the filtrate concentrated. Flash chromatography (silica, 57. CH30H/ethyl acetate) gavē the lactam M as a colorless crystalline solid. mp = 110-111eC; TLC Rf = 0.65 (20% CH3OH/ethyl acetate); 1H NMR (400 MHz, CDC13) δ 6.31 (bs.lH), 4.06 (m,2H), 3.31 (m,2H), 2.67 (m,3H), 2.28 (m,lH), 2.00-1.20 (m.llH), 1.45 (s,9H), 1.10 (m,2H).

Preparation of Ethyl [3(R)-[N-B0C-2-(Piperidin-4-yl)- ethvn-2-pjperidone-n acetate (49)_

To a stirred solution of 48 (6.7g, 22 mmol) and dry THF (150 mL) at -78*C was added NaN(TMS)2 (24.5 mL, 24.5 mmol, lM/hexanes) dropvise. After 15 min. ethyl bromoacetate (5.2 mL, 45 mmol) was added and then the reaction mixture warmed to 0*C for 1.0 h. The reaction was quenched with AcOH (1.0 mL) then diluted with ethyl acetate, washed with H2O and brine, dried (MgSO^), and concentrated. Flash chromatography (silica, 40% ethyl acetate/hexanes) gavē the ester 4J as a yellow oil. 88

TLC Rf = 0.26 (40% ethyl acetate/hexanes); NMR (300 MHz, CDCI3) δ 4.20 (q,j = 7 Hz,2H), 4.17 (d,J = 18 Ηζ,ΙΗ), 4.08 (m,2H), 3.98 (d,J = 18 Hz, 1H), 3.37 (m,2H), 2.68 (m,2H), 2.32 (m.lH), 2.00-1.25 (m,UH), 1.45(s,9H), 1.30 (t,J = 7 Hz,3H), 1.11 (m,2H).

Preparation of 3-(R)-[N-B0C-2-(Piperidin-4-yl)ethyl]- 2-pi-peridone-ll acetic acid (501)_ A solution of ϋ (6.0g, 15 mmol), 1N NaOH (50 raL, 50 mmol), and CH3OH (75 mL) was stirred at ambient temperature for 1.0 h. The reaction mixture was then acidified with 57. aqueous KHSO4 and then extracted with ethyl acetate. The organic portion was washed with brine, drjjed (MgS04&gt;, and concentrated to give the carboxylic acid 50 as a yellow oil. The oil was triturated with hexanes to give a fluffy white solid. TLC Rf = 0.31 (9:0.5:0.5 CH2C12/CH30H/Ac0H); XH NMR (400 MHz, CDC1) δ 4.13 (d,J = 17 Ηζ,ΙΗ), 4.07 (m,2H), 4.06 (d,J = 17 Ηζ,ΙΗ), 3.39 (m,2H), 2.33 (m, 1H), 1.95 (m,3H), 1.81 (m,lH), 1.70-1.25 (m, 7H), 1.45 (s,9H), 1.08 (m, 2H).

Preparation of 3(m-BOC-Amino-l-azobutan-2-one 151)

To a stirred solution of £1 (3.8 g, 20 mmol), 4-methylmorpholine (2.2 mL, 20 mmol), and ethyl acetate (200 mL) at -15*C was added isobutyl chloroformate (2.6 mL, 20 mmol). After l.Oh the reaction mixture was washed with H20 and brine, dried - 89 - LV10425 (MgSO^.), and filtered into a round bottom flask.

After cooling to 0eC the mixed anhydride was treated portion-wise with an ethereal solution of diazomethane &lt;80 mL, 40 mmol, δθ.5 M solution).

After 2.0 h the cooling bath was removed and the excess diazomethane removed by purging the solution with argon for 30 min. Concentration gavē the crude diazoketone 12. which was used directly for the next step. TLC Rf = 0.37 (307o ethyl acetate/hexanes).

Preparation of Ethvl N-BQC-3(R)-Methvl β-alanine 15.32 The crude diazoketone 12 &lt;4-. 3g, -20.0 mmol) was dissolved in ethanol (250 mL) and treated sequentially with NEt3 (3.4 mL, 24 mmol) and silver benzoate (1.4g, 6.0 mmol) to effect vigorous gas evolution and afford a black precipitate. After 1.0 h the reaction mixture was concentrated and the residue purified by flash chromatography (silica, 10% ethyl acetate/hexanes) to give the ethyl ester 12 as a colorless oil. TLC Rf = 0.42 (30% ethyl acetate/hexanes); XH NMR (400 MHz, CDC13) δ 4.96 (m.lH), 4.19 (q,J ,.= 7 Hz, 2H), 4.04 (m,lH), 2.52 (dd, J = 15 and 6 Hz, 1H), 2.46 (dd, J = 15 and 6 Hz, 1H), 1.44 &lt;s,9H), 1.26 &lt;t,J = 7 Hz,3H), 1.21 (d,J = 7 Hz, 3H). 90

Preoaration of Ethvl 3(R)-Methvl p-alanine?HCl_(54)

To a mechanically stirred solution of ϋ (2.2g, 9.7 mmol) in ethyl acetate (180 mL) at -15eC was vigorously bubbled HC1 gas for 30 min. The cooling bath (ethanol/ice) was removed and the solution purged with argon for 1.0 h to remove excess HC1. Concentration furnished the amineeHCl 54 as a yellow oil. ^H NMR (400 MHz, D20) δ 4.23 (q,J = 7 Hz,2H), 3.78 (m,1H), 2.79 (m,2H), 1.38 (d.J = 7 Hz, 3H), 1.29 (t,J = Hz,3H).

Preparātion of [3(R)-[N-B0C-2-(Piperidin-4-yl)ethyl]-2-piperidone-l]acetyl-3(R)-methyl~P-alanine ethyl ester (55)_

To a stirred solution 50. (1.0 g, 2.7 mmol), 54 (0.48 g, 2.8 mmol), HOBT (0.39g, 2.8 mmol), N(i-pr)2Et (1.5 mL, 8.5 mmol), and dry DMF (100 mL) at -15eC was added EDC (0.95 g, 2.8 mmol) followed by removal of the cooling bath. After 20 h the reaction mixture was diluted with ethyl acetate and then washed with H20, sat. NaHCO^, 5% aqueous KHSO4 and brine, dried (MgSO^.), and concentrated. Flash chromatography (silica, ethyl acetate) gavē 5i. as a colorless oil. TLC Rf = 0.35 (ethyl acetate); ^H NMR (300 MHz, CDCI3) 5 6.82 (bd, 1H), 4.32 (m, 1H), 4.13 (q, J = 7 Hz, 2H), 4.10 (d.J = 15 Hz, 1H), 4.08 (m,&quot;2H), 4.82 - 91 - LV10425 (d,J = 15 Hz, 1H), 3.36 (m,2H), 2.67 (m,2H), 2.48 (dd, J=5 and .1 Hz,2H), 2.33 2.00-1.20 (m,11H), 1.45 (s,9H), 1.27 (t,J = 7 Hz, 3H), 1.21 (d.J = 7 Hz,3H), 1.10 (m,2H).

Preparation of [3(R)-[N-B0C-2-(Piperidin-4-yl)ethyl]- 2-piperidone-nacetYl-3(R)-methvl-p-alanine (5.6J- A solution of JL5. (1-2 g, 2.5 mmol), 1N NaOH (10 mL, 10 mmol), and CH3OH (18 mL) was stirred at ambient temperature for 1.0 h. The reaction mixture was acidified with 57. aqueous KHSO4 and then extracted with ethyl acetate. The organic portion was then washed with brine, dried (MgSO^), and concentrated. Flash chromatography (silica, 10:0.5:0.5 ΟΗς^/ΟΗβΟΗ/ΗΟΑο) gavē the carboxylic acid as a colorless oil after azeotropic removal of residual HOAc with toluene. TLC Rf = 0.40 (10:0.5:0.5 CH2C12/CH30H/H0Ac); XH NMR (300 MHz, CDCI3) 57.02 (bd, ĪH), 4.35 (m, 1H), 4.12 (d, J = 16 Hz, 1H), 4.08 (m, 2H), 3.87 (d, J = 16 Hz, 1H), 3.32 (m, 2H), 2.69 (m, 2H), 2.56 (m, 2H), 2.36 (m. 1H), 2.00-1.25 (m, 11H), 1.45 (s, 9H), 1.03 (m, 2H) .

Preparation of [3(R)-[2-Piperidin-4-yl)ethyl]-2-piperidone-nacetvl-3(R)-methvl-ft-alanine (57) A solution of (0.74g, 1.6 mmol), trifluoroacetic acid (10 mL), and CH2C12 (10 mL) was stirred at ambient temperature for 1.0 h. The - 92 reaction mixture was then concentrated and the residual trifluoroacetic acid removed azeotropically with toluene. Flash chromatography (silica, 10:1:1 CH30H/NH40H/H20) afforded il as a white amorphous solid. Crystallization of £7 (100 mg) from hot ethanol (4.0 mL) gavē JLZ. (45 mg) as crystals after filtration at ambient temperature. 10 -L5 mp = 240°(D); TLC Rf = 0.48 (10:1:1 CH30H/NH40H/H20); J-H NMR (400 MHz, D20) 54.18 (m,lH), 4.06 (d,J = 16 Hz.lH), 3.95 (d,J = 16 Hz, 1H), 3.41 (m,4H), 2.99 (m, 2H), 2.45 (m.lH), 3.41 (m,4H), 2.99 (m,2H), 2.45 (in.lH), 2.42 (dd.J = 18 and 6 Hz, 1H), 2.33 (dd, J = 18 and 7 Hz, 1H), 1.97 (m, 4H), 1.90-1.55 (m,5H). 1.38 (m,4H), 1.19 (d,J = 7 Hz, 3H).

Preparation of l-[2-(N-Boc-Piperidin-4-yl)ethyl]-3- propen-2-vl-(15-tetrahvdropvrimidin-2-one) (59)_ 20 25

To a stirred suspension of lH-tetrahydropyri-midin-2-one (1.0 g, 10 mM) in DMF (50 ml) was added a 1.0 molar solution of lithium bis (trimethylsilyl)-amide (10 ml, 10 mM). After % hour, allyl bromide (1.0 ml, 1.17 mM) was added and stirring was continued for 1 hour. Asecond charge of lithium bis(trimethylsilyl)amide (10 ml, 10 mM) was added followed after % hour by the addition of (N-Boc-piperidin-4-yi) ethyl iodide (1(1). The reaction was stirred at 25eC overnight. The reaction mixture was poured into ice and 15% KHSO4 (50 ml), extracted with ether (2 x 100 ml) washed with H20 (2 x 100 ml), brine, dried over Na2S04 and evaporated ia vacuo. Flash chromatography on Si02 (ethyl acetate-hexane 30 - 93 - LV10425 1:1) afforded pure 53L. ΧΗ NMR (300 MHz, CDCI3) δ 1.14 (2H, m), 1.46 (9H, s), 1.74 (3H, m), 1.95 (2H, m) 2.68 (2H, t), 3.22 (4H, m), 3.38 (2H, m), 3.95 (2H, m), 4.06 (2H, m), 5.13 (2H, dd).

Preparation of l-[2-(N-Boc-Piperidin-4-yl)ethyl]-3-acetaldehvde-(lH-tetrahvdroDvrimidin-2-one) (60)

To a stirred solution of 5J2. (430 mg, 1.27 mM) in THF (7 ml), H2O (5 ml) and sodium periodate (0.68 g, 3.19 mM) was added a 2.5% solution of osmium tetroxide in 2-methyl-2-propanol (413 μΐ, 1.28 mM). After stirring for 4 hours, ethyl acetate (70 ml) was added and the mixture vashed with 50% brine/10% sodium sulfate (1:1) then with brine. The solution was dried over Na£S04 and evaporated in vacuo to give &amp;2.

Preparation of l-[2-(N-Boc-Piperidin-4-yl)ethyl]-3- acetic acid-(lH-tetrahvdropvrimidih-2-one) ,(,61)- A solution of ££ (374 mg) in acetone (7 ml) was cooled to -15eC then treated dropvise with Jones reaģent until an orange color persisted for 5 minūtes. Ethyl acetate (60 ml) was added and the solution washed with H20, brine, dried over Na2S04, then evaporated in vacuo to give £1. 1H NMR (300 MHz, CDCI3) δ 1.12 (2H, m), 1.45 (9H, s), 1.70 (2H, bd), 2.03 (2H, m), 2.70 (2H, bt), 3.37 (4H, m), 3.95 (2H, m), 4.05 (2H, m). 94

Preparation of [l-[2-(N-Boc-Piperidin-4-yl)ethyl]-(lH-tetrahydropyrimidin-2-one)-3]-acetyl-3(R)-(2- phenvlethvl)J3-alaninemethvlester (63 ^ _ A mixture of 61 (125 mg, 0.34 mM), (R)-3-(2-phenylethyl)B-alanine methyl ester hydrochloride (83 mg, 0.34 mM) (61), l-hydroxybenzotriazole hydrate (48.3 mg, 0.35 mM), l-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride (70 mg, 0.37 mM) and triethyl amine (107 μΐ, 0.77 mM) in dry DMF (1.6 ml) was stirred at 25eC for 18 hours. The reaction mixture was poured into ice Η£θ, extracted with ethyl acetate (2 x 50 ml), washed with 10% citric acid, brine, dried over Na2S04 and evaporated in vacuo to give 61. XH NMR (300 MHz, CDCI3) δ 1.14 (2H, m), 1.45 (9H, s), 1.70 (2H, m), 1.86 (2H, m), 2.02 (2H, m), 2.58 (2H, d), 2.66 (3H, m), 3.36 (5H, m), 3.68 &lt;2H, m), 3.93 (1E, d), 4.07 (2H, m), 7.02 (1H, m), 7.20 (2H, m), 7.29 (2H, m).

Preparation of l-[2-(N-Boc-Piperidin-4-yl)ethyl]-(lH-tetrahydropyrimidin-2-one)-3]-acetyl-3(R)-(2- phenvlethvl)J3-alanine (64)__ A solution of 61 (168 mg, 0.31 mM) in THF (15 ml) and H2O (15 ml) was cooled to 0*C, treated with IN LiOH (1.2 ml) then stirred at 25eC overnight. The THF was evaporated at reduced pressure, the residue acidified with 10% citric acid, extracted with ethyl acetate (2 x 50 ml) washed with brine, dried over Na2SŪ4 and evaporated iņ vacuo to give 64. - 95 - LV10425

Preparation of [l-(2-piperidin-4-yl)ethyl]-[(lH-tetrahydropyrimidin-2-one)-3]-acetyl-3(R)-(2-phenyl- ethyl)flralanine (65)- A solution of M &lt;128 mg) in meth^lene chloride (8 ml) and anisole (1.5 ml) was cooled to -20eC then treated with trifluoroacetic acid (8 ml) over a 5 minūte period. The reaction mixture was stirred at -5*C for 1 hour then evaporated in vacuo. The residue was chromatographed on S1O2 (ethanol-^O-NH^OH, 10:1:1) to give 61-Analysis Calc for C24H36N4°4*^.5^0 C, 57.84; H, 8.65; N, 11.73.

Found: C, 57.94; H, 8.39; N, 11.47.

Preparation of [l-(2-piperidin-4-yl)ethyl]-[(lH-tetrahydropyrimidin-2-one)-3]-acetyl-3(R)-(2-indol- 3-vl)-ethvlU3-alanine (66)_

By folloving substantially the procedure described for preparing 61 but substituting for the 3(R)-(2-phenylethyl)Ū-alanine methyl ester hydro-chloride described therein an equivalent amount of 3R-[2-(indol-3-ethyl]8-alanine ethyl ester and folloving the procedures described for preparing 64 and 61, there is obtained 66.

Analysis Calc for ^26^37^5041 C, 56.3; H, 8.0; N, 12.6.

Found: C, 56.3; H, 7.9; N, 12.2. 96

Preparation of [l-(2-piperidin-4-yl)ethyl-(lH-tetrahydropyrimidin-2-one)-3]-acetyl-3(R)-methyl β- alanine (67)____

By folloving substantially the procedure described for preparing 6i but substituting for the (R)-3-(2-phenylethyl)B-alanine methyl ester hydro-chloride described therein an equivalent amount of ethyl 3(R)-methylfi-alanine hydrochloride and folloving the procedures described for preparing 64 and 6i, there is obtained 67.

Preparation of [l-(2-piperidin-4-yl)ethyl]-[(lH-tetrahydropyrimidin-2-one)-3]-acetyl-2(R)-[butyl- sulfonvlaminol B-alanine (68)_

By folloving substantially the procedure described for preparing 6i but substituting for the (R)-3-(2-phenylethyl)B-alanine methyl ester hydro-chloride described therein an equivalent amount of ethyl 2(S)-butylsulfonylamino β-alanine hydrochloride and folloving the procedures described for preparing 64 and 6i. there is obtained M·

Preparation of Methyl 4(R)-Benzyl-2-oxazolidinone- SUCCin^tg- L7.0.Ž__

To a stirred solution of mono-methyl succinate 62. (3.7 g, 28.2 mmol), NEt3 (4,0 mL, 28.2 mmol) and dry THF (160 mL) at -78eC vas added trimethylacetyl chloride (3.6 mL, 28.2 mmol) dropvise. After 10 min, the cooling bath vas removed and replaced vith an icebath. After 1.0 hour, the heterogeneous mixture vas recooled to -78°C, and treated, via cannula addition, vith lithium (R)-(+)-4 benzyl-2-oxazolidinone (28.2 mmol) in dry THF (43 mL), prepared by treating (R)-(+)-4-benzyl-2- - 97 - LV10425 oxazolidinone (5.0 g, 28.2 mmol) in dry THF (43 mL) at -78eC with n-BuLi (17.8 mL, 28.2 mmol, 1.6 M/hexanes). After addition was complete, the reaction mixture was warmed to room temperature overnight. The reaction mixture was diluted with ethyl acetate and washed with H20, sat. NaHC03, 5% aqueous KHSO4 and brine, dried MgSŪ4 and concentrated. Flash chromatography (silica, 207. ethyl acetate/hexanes) gavē Zfi. as a white crystalline solid. TLC Rf=0.48 (307. ethyl acetate/hexanes) ; XH NMR (300 MHz, CDCI3) δ 7.30 (m, 5H), 4.68 (m, 1H), 4.21 (m, 2H), 3.72 (s, 3H), 3.28 (m, 3H), 2.80 (dd, J=13 and 4Hz, 1H), 2.73 (m, 2H).

Preparation of Methyl 4(R)-Benzyl-2-oxazolidinone- 2(R)-(2-cvanoethvl)succinate (71)-

To a stirred solution of T1CI4 (12.3 mL, 12.3 mmol, 1M/CH2C12) and dry CH2C12 (83 mL) at 0eC was added titanium (IV) isopropoxide (1.2 mL, 4.1 mmol). After 15 min, diisopropylethylamine (3.0 mL, 17.3 mmol) was added dropwise to form a dark brovm solution. After 10 minūtes, Z£L (4.8 mg, 16.5 mmol) was added followed by continued stirring at 0*C for 1 hour. Acrylonitrile (4.3 mL, 66 mmol) was added dropvise to the dark solution followed by stirring at 0eC for 72 hours. The reaction mixture was quenched with sat. NH4CI at 0*C, then extracted with CH2C12.

The combined organic extracts were washed with saturated NaHC03 and brine, dried MgS04 and concentrated. Flash chromatography (silica, 25% ethyl acetate/hexanes) gavē 71 as a crystalline solid. TLC Rf=0.22 (30% ethyl acetate/hexanes); 98 XH NMR &lt;300 ΜΗζ, CDCI3) δ 7.40-7.20 (m, 5Η) , 4.70 (m, 1Η), 4.25 (m, 2H), 4.13 (m. 1H), 3.67 (s, 3H), 3.30 (dd, J=13 and 4Hz, 1H), 2.93 (dd, J=17 and 10Hz, 1H), 2.80 (dd, J=17 and 10Hz, 1H), 2.55 (dd, J=13 and 4Hz, 1H), 2.43 (t, J=7Hz, 2H), 2.10 (m, 1H), 1.90 (m, 1H&gt;.

Preparation of Methyl 4(R)-Benzyl-2-oxazolidinone- 2('R)-(3-aminopropvl)sticcinate (72)_ A mixture of IX (2.0 g, 5.8 mmol), Pt02 (0.8 g), CH3OH (50 mL), and CHCI3 (5 mL) were shaken on the Parr apparatus under a hydrogen atmosphere (60 PSI) at ambient temperature for 3 hours. The reaction mixture was filtered through a celite pad and then concentrated to furnish the crude amine*HCl 12. as a yellow oil. ~~ TLC Rf=0.50 (CH2C12/CH30H/Ac0H 9:1:1)

Preparation of Methvl r2-Pineridone-3(R)lacetate (731 The crude amine«HCl 12 (2.0 g, ~5.8 mmol), acetonitrile (250. mL) and NaHC03 (5 g) were stirred overnight at ambient temperature. The reaction mixture was then filtered and the filtrate concentrated. Flash chromatography (silica, ethyl acetate) gavē lactam 12 as a colorless crystalline solid. TLC Rf=0.22 (ethyl acetate); XH NMR (300 MHz, CDCI3) δ 5.82 (bs, 1Η), 3.70 (s, 3H), 3.33 (m, 2H), 2.88 (dd, J=16 and 4Hz, 1H), 2.75 (ra, 1H), 2.58 (dd, J=16 and 7Hz, 1H), 2.10-1.60 (m, 4H) . - 99 - LV10425

Preparation of Methyl [l-[N-Boc-2-(Piperidin-4-yl)~ ethvn-2-piperidone-3(R)1acetate (7A)_

To a stirred solution of 73. (1.1 g, 6.4 nunol) in anhydrous DMF (60 mL) at -15eC was added NaN(TMS)2 (6.4 mL, 6.4 mmol, lM/hexanes) dropwise folloved by removal of the cooling bath. After 10 minūtes the iodide lfi. (2.3 g, 7.0 mmol) was added and stirring continued at ambient temperature for 4 hours. The reaction mixture was then diluted with ethyl acetate and washed with H20 (2x) and brine, dried (MgS04&gt;, and concentrated. Flash chromatography (silica, 40% ethyl acetate/hexanes) gavē 24 as an oil. TLC Rf=0.29 (40% ethyl acetate/hexanes).

Preparation of [l-[N-Boc-2-(piperidin-4-yļ)ethyl]-2- piperidone-3(R)1acetic acid_(75)_

Utilizing the procedure for converting 42. to 50. 74 (0.44 g, 1.1 mmol) furnished 22 as an oil. TLC Rf=0.52 (CH2C12/CH3OH/AcOH 9:0.5:0.5).

Preparation of [l-[N-Boc-2-(piperidin-4-yl)ethyl]-2-piperidone-3(R)]acetyl-3(R)-methyl-B-alanine ethyl ester (76)_

Utilizing the procedure for converting to 55. 22 (0.40 g, 1.1 mmol) was coupled to 24. (0.27 g, 1.6 mmol) to afford JA as an oil after flash chromatography (silica, ethyl acetate). TLC Rf=0.20 (ethyl acetate). 100 -

Preparation of [l-[N-(Boc-2-(piperidin-4-yl)ethyl]-2-piperidone-3(R)1acetvl--3(R)-methvl-fl-alanine (77) Utilizing the procedure for converting 5i. to 56. 76 (0.30 g, 0.64 mmol) gavē H as an oil. TLC Rf=0.24 (CH2C12/CH30H/Ac0H 9:0.5:0.5); 1Έ. NMR (400 MHz, CBCI3) 5 6.93 (m, 1H), 4.32 (m, 1H), 4.06 (m, 2H), 3.35 (m, 2H), 3.29 (m, 2H), 2.70-2.50 (m, 6H), 2.38 (m, 1H), 2.00-1.20 (m, 9H), 1.45 (s, 9H), 1.27 (d, J=6Hz, 3H), 1.06 (m, 2H).

Preparation of [l-[2-(Piperidin-4-yl)ethyl]-2- pineridone-3(Rl-roethvl-fl-alanine (78)_

Utilizing the procedure used to convert 5i. to 57. 77_ (0.24 g, 0.53 mmol) gavē 7&amp; as an amorphous solid āfter flash chromatography (silica, 10:1:1 ethanol/H20/NH40H). ^ NMR (400 MHz, D20) δ 4.19 (m, 1H), 3.40 (m, 6H), 3.00 (m, 2H), 2.70 (m, 1H), 2.64 (m, 1H), 2.47 (m, 2H), 2.30 (dd, J=14 and 8Hz, 1H), 2.00 (m, 2H), 1.92 (m, 2H), 1.79 (m, 1H), 1.61 (m, 4H), 1.45 (m, 2H), 1.18 (d, J=7Hz, 3H).

Preparation of N-Cbz-Gly-3(R)-methyl-B-alanine ethyl ester (79)___ A stirred solution of Cbz-Gly (0.94 g, 4.5 mmol), 4-methylmorpholine (1.1 mL, 4.9 mmol), and ethyl acetate (50 mL) at -15 *C was treated with isobutyl chloroformate (0.61 mL, 4.7 mmol) in one portion. After 15 min, 5A (0.75 g, 4.5 mmol) in ethyl acetate (1 mL) was added followed by warming to ambient temperature overnight. The reaction mixture was then washed with 1M NaHSO^, brine, sat. NaHC03 and brine, dried (MgSO^, and concentrated to afford 7i as a yellow oil. - 101 - LV10425 NMR (300 MHz, CDCI3) 6 7.48 (m, 5H), 6.73 (m, 1H), 5.58 &lt;m, 1H), 5.13 (s, 2H), 4.48 (m, 1H), 4.30 &lt;q, J=7Hz, 2H), 3.95 (m, 2H), 2.62 (m, 2H), 1.37 (t, J=7Hz, 3H).

Preparation of Gly-3(R)-methyl-B-alanine ethyl ester acetic acid salt (80)_ A mixture of 23. (1.4 g, 4.5 mmol), 107. AcOH/ CH3OH (50 mLJ, and 207» Pd(OH&gt;2 (0.14 g) was shaken on the Parr apparatus under a hydrogen atmosphere (60 PSI)'overnight. After 24 hours the reaction mixture was filtered through a celite pad and the filtrate concentrated. Flash chromatography (silica, 5:3:1 CH2Cl2/€thanol/AcOH) to give ££. as an amorphous solid. TLC Rf=0.41 (5:3:1 CH2Cl2/ethanol/AcOH).

Preparation of 4(S)-Benzyl-2-oxazolidinone-4-(N-Boc- piperidin-4-vl)-2(RJ-(prap-2-enfi-)butvtate (8.1.)-

To a stirred solution of 45. (2.5 g, 5.8 mmol) in THF (50 ml) at -78eC was added lithium bis(trimethylsilyl)amide (7.0 mL, 7.0 mmol, lM/hexanes) folloved by allyl bromide (2.5 mL, 29 mmol). The cooling bath was then removed and the reaction stirred at 0eC for 1.5 hours. The reaction was quenched with sat. NH4CI, then diluted with EtOAc followed by washing with sat. NaHCC^, 57. KHSO4, and brine, drying (MgSŪ4) and concentration. Flash chromatography (silica, 15% EtOAc/hexanes) afforded 81 as a colorless oil. TLC Rf=0.53 (307. EtOAc/hexanes ) ; 102 XH NMR (300 ΜΗζ, CDC13) δ 7.40-7.20 (m, 5Η), 5.82 (m, 1Η), 5.10 (m, 2H), 4.80 (m, 1H), 4.20 (m, 2H), 4.07 (m, 2H), 3.90 (m, 1H), 3.31 (dd, J=13 and 3Hz, 1H), 2.68 (m, 2H), 2.48 (m, 1H) , 2.35 (m, 1H), 1.90-1.20 (m, 7H), 1.47 (s, 9H), 1.10 (m, 2H). *

Preparation of 4-(N-Boc-Piperidin-4-yl)-2(R)-(prop-2- ene)butvric acid (_82J._ -—

To a stirred solution of !1 (1.8g, 3.8nunol), 30% H202 (8.5 mL, 83 mmol), THF (41 mL) and H20 (12mL) at ambient temperature was added LiOH (14 mL, 14 mmol, 1N). After 2h the excess LiOH was guenched with 10% NaHSŪ4 dropvise at 0eC. The reaction was then acidified with 57. KHSO4 and extracted with EtOAc. The EtOAc portion was then washed with brine, dried (MgS04&gt;, and concentrated to give 12. as a colorless oil. TLC Rf = 0.79 (10% CH3OH/EtOAc).

Preparation of [4-(N-Boc-Piperidin-4-yl)-2(R)-(prop-2-ene)butyrate]-Gly-3(R)-methyl-B-alanine ethyl ester M)_

Utilizing the procedure for converting ϋ to 55. 82 (1.3 g, 2.5 mmol) was coupled to 10. (0.62 g, 2.5 mmol) to give H after flash chromatography (silica, 107. isopropanol/hexanes). TLC Rf=0.50 (20% isopropanol/hexanes); XH NMR (300 ΜΗζ, CDC13) δ 6.67 (bd, J=8Hz, 1H), 6.37 (m, 1H), 5.72 (m, 1H), 5.02 (m, 1H), 4.35 (m, 1E), 4.13 (q, J=7Hz, 2H), 4.03 (m, 1H), 3.88 (m, 2H), 2.63 (m, 2H), 2.51 (m, 2H), 2.35 (m, 1H), 2.20 (m, 2E), 1.70-1.20 (m, 7H), 1.44 (s, 9H), 1.26 (t, J=7Hz, 3H), 1.07 (m, 2H). - 103 - LV10425

Preparation of [3-[2-(N-Boc-Piperidin-4-yl)ethyl]-5-hydroxy-2-pyrrolidinone-l]acetyl-3(R)-methyl-B-alanine ethvl ester (84)-

To a solution of §1 (0.74 g, 1.5 mmol), THF (12.2 mL), H20 (9.2 mL), and NaI04 (0.82 g, 3.8 mmol) was added 0s04 (0.96 mL, 80 μπιοί; 2.57. tert-butanol) at ambient temperature. After 2.0 hours, the reaction mixture was diluted with ether and then washed with H20, 107. aqueous Na2S203, and brine, dried (MgS04), and concentrated to give M as a yellow oil. TLC Rf=0.26 (207. isopropanol/hexanes) .

Preparation of [3-[2-(N-Boc-Piperidin-4-yl)ethyl]-5-hydroxy-2-pyrrolidinone-l]acetyl-3(R)-methyl-B- alanine (85)______

Utilizing the procedure for converting ϋ to 56. 84 (0.73 g, 1.5 mmol) gavē £j&gt;. as a pale yellow foam. *Η NMR (300 MHz, CDC13) δ 5.23 (d, J=5Hz, 1H), 4.58 (d, J=16Hz, 1H), 4.32 (m, 1H), 4.05 (m, 2H), 3.70 (d, J=16Hz, 1H), 2.80-2.40 (m, 6H), 2.22 (m, 1H), 2.00-1.00 (m, 9H), 1.45 (s, 9H), 1.27 (d, J=7Hz, 3H).

Preparation of [3-[2-(Piperidin-4-yl)ethyl]-2-pyrrol- idinone-11acetyl-3(R)-methvl-8-alanine (86)_ A stirred solution of gū. (0.61 g, 1.3 mmol) in CH2C12 (50 mL) at ambient temperature was treated with a solution of CF3C02H (5.8 mL) and triethylsilane (0.8 mL, 5.2 mmol). After 2.0 h the reaction mixture was concentrated folloved by azeotropic removal of residual 0Γ3002Η with toluene. Flash chromatography (silica, 10:1:1 ethanol/H20/NH40H) gavē 86 as a colorless solid. - 104 - TLC Rf=0.17 (9:1:1 ethanol/^O/NH^OH) ; 1H NMR (300 MHz, CD3OD) δ 4.20 (m, 1H), 3.97 (d, J=17Hz, 1H), 3.85 (d, J=l7Hz, 1H), 3.40 (m, 4H), 2.95 (m, 2H), 2.52 (m, 13), 2.30 (m, 23), 2.00-1.30 (m, 11H), 1.20 (d, J=7Hz, 3H).

Preparation of 2-(2,3,4,5,6-Tetrahydro-l,l-dioxo- 2H-1.2-thiazin-2-vl)prop-2-ene (8ΒΛ-

To a solution of jjLZ. (J. 0. C. Ū2, p. 2162 (1987) E.H. White, H.M. Lim) (0.35 g, 2.6 mmol) in DMF at 0eC was added NaN(TMS&gt;2 (3.9 mL, 3.9 mmol); lM/hexanes) dropwise. After 30 min, allyl bromide (0.45 mL, 5.2 mmol) was added in one portion followed by continued stirring for 30 min. The reaction mixture was diluted with ethyl acetate and washed with sat. NH^Cl and brine, dried (MgSC^), and concentrated. Flash chromatography (silica, 307. ethyl acetate/hexanes) gavē 88 as a colorless oil. TLC Rf=0.29 (307. ethyl acetate/hexanes); 1H NMR (300 MHz, CDCI3) δ 5.80 (m, 1H), 5.25 (m, 2H), 3.78 (m, 2H), 3.30 (m, 2H), 3.02 (m, 2H), 2.21 (m,' 2H), 1.68 (m, 2H).

Preparation of 2-[6-[2-(N-Boc-Piperidin-4-yl)ethyl]-(3,4,5,6-tetrahydro-l,l-dioxo-2H-l,2-thiazin-2-yl)]- prop-2-ene (89)_ A stirred solution of M (0.34 g, 1.93 mmol) in THF (10 mL) at -78°C was treated dropwise with n-BuLi (1.45 mL, 2.3 mmol; 1.6 M/hexanes). After 30 min the reaction mixture was treated sequentially with l-methyl-2-pyrrolidinone (1 mL) and 10 (0.86 g, 2.7 mmol) in THF (10 mL), followed by warming&quot;to -23®C. After 2 hours at -23°C the reaction was quenched with - 105 - LV10425 saturated NH4C1 then diluted with ethyl acetate and washed with H2O and brine, dried (MgS04), and concentrated. Flash chromatography (silica, 307. ethyl acetate/hexanes) gavē £2. as a crystalline solid. ,.TLC Rf=0.27 (30% ethyl acetate/hexanes); ΧΗ NMR (400 MHz, CDCI3) δ 5.80 (m, 1H), 5.25 (m, 2H), 4.07 (m, 2H), 3.78 (m, 2H), 3.34 (m, 2H), 3.17 (m, 1H), 3.02 (m, 1H), 2.82 (m, 2H), 2.66 (m, 2H), 2.24- 2.04 (m, 3H), 1.89 (m, 1H), 1.70-1.35 (m, 7H), 1.08 (m, 2H).

Preparation of 2[6-[2-(N-Boc-Piperidin-4-yl)ethyl]-(3,4,5,6-tetrahydro-l,l-dioxo-2H-l,2-thiazin-2-yl)]- acetaldehvde (90)__

To a stirred solution of 82. (0.50 g, 1.29 mmol), THF (16 mL), H20 (12 mL), and NaI04 (0.69 g, 3.2 mmol) was added 0s04 (1 mL, 0.1 mmol; 2.5 %/tert-butanol) to effect a white precipitate. After 3.0 h, the reaction mixture was diluted with ethyl acetate and washed with H20, 5% KHS04, sat. NaHC03 and brine, dried (MgS04), and concentrated to give aldehyde 211 as an oil. 1H NMR (400 MHz, CDC13) δ 9.64 (s, 1H), 4.08 (m, 2H), 4.00 (d, J=17Hz, 1H), 3.89 (d, J=17Hz, 1H), 3.62 (m, 1H), 3.14 (m, 1H), 2.96 (m, 1H), 2.66 (m, 2H), 2.20 (m, 1H), 2.08 (m, 1H), 2.00-1.35 (m, 9H), 1.10 (m, 2H) .

Preparation of 2-[6-[2-(N-Boc-Piperidin-2-yl)ethyl]-(3,4,5,6-tetrahydro-l,l-dioxo-2H-l,2-thiazin-2-yl)]- acetic acid (91)___ A stirred solution of 2ϋ (0.44 g, 1.13 mmol) in acetone (10 mL) at 0eC was treated with Jones Reaģent (1.0 mL) in 3 portions. After 5 min, excess - 106 -

Jones Reaģent was queiiched with i-propanol and then diluted with EļO. The reaction mixture was then extracted with ethyl acetate and the organic extract washed with brine, dried (MgSO^.), and concentrated. Flash chromatography (silica, 20:1:1 CH2CI2/CH3OH/ AcOH) gavē as a light yellow oil. TLC Rf=0.45 (20:1:1 CH2CI2/CH3OH/ACOH).

Preparation of 2-[6-[2-(N-Boc-Piperidin-4-yl)ethyl]-(3,4,5,6-tetrahydro-l,l-dioxo-2H-l,2-thiazin-2-yl)]- acetvl-3(R)-methvl-B-alanine ethvl ester (92.)---

Utilizing the procedure for converting 20. to 55. 91 (98 mg, 0.24 mmol) was coupled to 14 (120 mg, 0.71 mmol) to afford 22. as a yellow oil after flash chromatography (silica, ethyl acetate). TLC Rf=0.28 (ethyl acetate); % NMR (300 MHz, CDCI3) S 7.08 (m, 1H), 4.37 (m, 1H), 4.08 (m, 2H), 3.89 (d, J=17Hz, 1H), 3.77 (d, J=17Hz, 1H), 3.60 (m, 1H), 3.15 (m, 1H), 2.88 (m, 1H), 2.68 (m. 2H), 2.52 (m, 2H), 2.20-1.00 (m, 13H), 1.27 (t, J=7Hz, 3H), 1.25 (t, J=7Hz, 3H).

Preparation of 2-[6-[2-(N-B0C-Piperidin-4-yl)ethyl]-(3,4,5,6-tetrahydro-l,l-dioxo-2H-l,2-thiazin-2-yl)]- acetvl-3(R)-methvl-B-alanine (93) ___

Utilizing the procedure for converting 2i to 56. 92 (76 mg, 0.15 mmol) gavē 23. as an oil. TLC Rf = 0.42 (20:1:1 CH2C12/CH30H/AcOH); J-H NMR (300 MHz, CDCI3) δ 7.20 (m, 1H), 4.39 (m, 1H), 4.10 (q, J=7 Hz, 2H), 4.08 (m, 2H), 3.92 (d, J=17 Hz, 1H), 3.74 (m, 1H), 3.53 (m, 1H), 3.15 (m, 1H), 2.90 (m, 1H), 2.77 (m, 2H), 2.59 (m, 2H), 2.10 (m, 1H), 2.10-1.10 (m, 12H), 1.27 (t, J=7 Hz, 3H), 1.25 (t,

Hz, 3H). - 107 - LV10425 ι·

Preparation of 2-[6-[2-(Piperidin-4-yl)ethyl]-(3,4,5,-6-tetrahydro-l,l-dioxo-2H-l,2-thiazin-2-yl)]acetyl-3- (R)-methvl-fi-alanine (94).________

Utilizing the procedure for converting ££. to 57. 93 (61 mg, 0.12 mmol) gavē £4 as an off-white solid after flash chromatography (silica, 10:1:1 ethanol/NH40H/H20). TLC Rf = 0.26 (10:1:1 ethanol/HH40H/H20); NMR (300 MHz, D20) δ 4.08 (m, 1H), 3.71 (m, 2H), 3.40 (m, 1H), 3.29 (m, 2H), 3.14 (m, 2H), 2.83 (m, 2H), 2.25 (m, 2H), 2.12 (m, 1H), 1.90-1.20 (m, 12H), 1.05 (d, J=7 Hz, 3H). 1,=12-(N-B0C-piperidin-4-yl)ethyl]-3-propanol-2-pyrrol- idinone (95)__—--- A solution of 20 (1.20 g, 3.6 mmol) in THF (3 ml) was added to a solution of borane-dimethyl sulfide (3.6 īrnol) in 3 ml THF at 0eC and this was stirred for 2.5 hr at 0·. Then, H20 (0.15 ml), 10N aqueous NaOH solution (0.65 ml), THF (1 ml), and C2H50H (0.36 mL) were added at 0e and with stirring the reaction mixture was warmed to 23° and treated with 307. H202 (0.36 ml). This solution was heated to 55eC and stirred for 2 hours. The reaction mixture was cooled, saturated with Κ2002 an the organic phase was separated, dried (MgS04) and the solvent removed. The resulting residue was purified by flash chromatography on silica gel eluting with 107. MeOH/EtOAc to give £5.. Rf 0.3 (silica gel, 107. MeOH/EtOAc. 108 - l_[2-(N-B0C-piperidin-4-yl)ethyl]-3-propanoic-acid- 2-pvrrolidinone (96)___ A solution of 25. (1.14 mmol) in acetone (5 ml) was cooled to -15* and treated with Jones reaģent until the color remained amber/brown. The reaction mixture was diluted with saturated NaHCOg solution to pH 9 and this was extracted with EtOAc. The aqueous phase was acidified to pH 3 with KHSO^ solution and extracted with several portions of EtOAc. The combined organic extracts were dried (MgSO^) and the solvent removed to give 22· Rf 0.3 (silica gel, 9/1/1 CH2Cl2/MeOH/HOAc).

[l-[2-(N-B0C-piperidin-4-yl)ethyl]-2-pyrrolidinone- 31propanovl-J3-alanine ethvl ester (97)_ A solution of 22 (0.55 mmol), β-alanine ethyl ester*HCl (0.825 mmol), N-methylmorpholine (1.65 mmol) in CH3CN (5 ml) at 23° was treated with BOP (0.825 mmol) and the resulting solution was stirred for 48 hrs. The reaction mixture was diluted with EtOAc, washed with 10% KHSO4, brine, H20, brine, dried (MgSO^) and the solvent removed. The residue was purified by flash chromatography on silica gel eluting with 307. acetone/hexane to give 21 as a clear oil. Rf 0.25 (silica gel, 30% acetone/hexane).

[l-[2-(N-B0C-piperidin-4-yl)ethyl]-2-pyrrolidinone- 31propanovl-Q-alanine (98)_ A solution of 22 (0.29 mmol), LiOH*H20 (1.45 mmol), THF/Me0H/H20 (1:1:1, 3 ml) was stirred for 2 hours. This was diluted with EtOAc and washed with 107. KHSO^, H20, brine and dried (MgSO^.). Solvent removal gavē 28 as an oil. Rf 0.5 (silica gel, 9/1/1 CH2Cl2/Me0H/H0Ac). - 109 - LV10425 [l-[2-(Piperidin-4-yl)ethyl]-2-pyrrolidinone-3]pro- panovl-B-alanine *HC1 (99)_ A solution of 22 (0.18 nunol) in EtOAc (2 ml) was cooled to -78· and treated with EC1 gas. After 0.5 hr the solvent was removed to give 22 as a white solid. Rf 0.22 (silica gel, 10/1/1 EtOH,'NH40H, H20).

Preparation of N-B0C-4-piperidine methvl iodide (101) Utilizing the procedure for converting 2 to 12, 122 &lt;12.3 g, 57 mmol) (Carr et al., EP 317997, May 31, 1989), gavē 101 after flash chromatography (silica gel, 15% EtOAc/hexane). Rf 0.38 (silica gel, 10%

EtOAc/hexanes). XH NMR (300 MHz, CDCI3) 5 1.15 (2H, m), 1.48 (9H, s), 1.62 (1H, m), 1.84 (2H, m), 2.70 (2H, m), 3.11 (2H. m), 4.12 (2H, m).

Preparation of N-BOC-piperidinemethyltriphenylphos- phonium iodide (102)_ A solution of 101 (5.1 g, 15.7 mmol), CH3CN (75 mL), and triphenylphosphine (4.5 g, 17.3 mmol)-was heated at 80*C for 60 h. The cooled reaction mixture was concentrated and the residual oil triturated with ether, then placed under vacuum to give 102 as a yellow foara. 1H NMR (CDCI3, 300 MHz) δ 7.31 (m, 15H), 4.10 (m, 2H), 3.97 (m, 2H), 2.69 (m, 2H), 2.00-1.10 (m, 5H), 1.48 (s, 9H). 110 -

Preparation of 3-[2-(N-B0C-piperidin-4-yl)ethylene]-2- (methoxv)pyridine (104)_

To a suspension of 102 (5.0 g, 8.5 mmol) in dry THF at 0eC was added NaN(TMS)2 &lt;9.4 mL, 9.4 mmol, lM/hexanes) dropvise. After 15 min. a solution of 103 (0.97 g, 7.0 mmol) (Tetrahedron Letters (1988) 2i, 773) in THF (3 mL) was added dropvise to the yellow/orange homogeneous reaction mixture. After lh the cooling bath was removed folloved by continued stirring for 30 min. The reaction was quenched with H20 and ether. The aqueous phase was reextractedwith ether, the organic layers combined and then washed with brine, -dried (Na2S0^) and concentrated. Flash chromatography (silica, 15% ethyl acetate/hexanes) gavē 104 as a colorless oil. — TLC Rf = 0.15 (10% ethyl acetate/hexanes), ^ NMR (300 MHz, CDCI3) δ 8.09 (dd, J=5 and 2 Hz, 1H), 7.42 (dd, J=7 and 2Hz, 1H), 6.87 (dd, J=7 and 5 Hz, 1H), 6.37 (d, J = 12 Hz, 1H), 5.58 (dd, J=12 and 10 Hz, 1H), 4.10 (m, 2H), 3.95 (s, 3H), 2.70 (m, 2H), 2.50 (m, 1H), 1.80-1.20 (m, 4H), 1.47 (s, 9H).

Preparation of 3-C2-(N-B0C-piperidin-4-yl)ethyl]-2- roethoxvpvridine (105)_ A mixture of 104 (1.4 g, 4.3 mmol), ethyl acetate (22 mL) and 10% Pd/C (0.27 g, 20%/wt) was stirred under a hydrogen atmosphere (1 atm) at ambient temperature for 4 h. The reaction mixture was filtered through a celite pad and the filtrate concentrated to give 105 as a cloudy colorless oil. - 111 - LV10425 TLC Rf = 0.17 (107. ethyl acetate/hexanes); NMR (CDCI3, 300 MHz) δ 8.02 (dd, J=5 and 2 Hz, 1H), 7.36 (dd, J=7 and 2 Hz, 1H), 6.80 (dd, J=7 and 5 Hz, 1H), 4.10 (m, 2H), 3.94 (s, 3H), 2.69 (m, 2H), 2.58 (m, 2H), 1.72 (m, 2H), 1.55-1.35 (m, 3H), 1.48 (s, 9H), 1.13 (m, 2H).

Preparation of 3-[2-(N-B0C-piperidin-4-yl)ethyl]-2- pyridon-l-yl (106)__

To a solution of 105 (0.81 g, 2.5 mmol) in dry CH3CN (13 mL) was added Nal (0.94 g, 6.3 mmol) folloved by chlorotrimethylsilane (0.80 mL, 6.3 mmol). The resulting opaque yellow solution was heated.at 60eC for 2.0 h. The cooled reaction mixture was quenched with methanol (50 mL), stixred for 10 min. and then concentrated. The residue was dissolved in dioxane/H20 (6.7 mL/3.3 mL) then treated with 1N NaOH (5.0 mL, 5.0 mmol). The solution was cooled to 0eC and then treated with BOC2O (1.1 g, 5.0 mmol) folloved by removal of the cooling bath. After stirring overnight the dioxane was evaporated and -the residue diluted with H2O (10 mL) and ethyl acetate (50 mL), followed by acidification to pH - 1.0 with 207. KHSO4. The organic phase was separated and then washed with H2O and brine, dried (MgSO^) and concentrated. Flash chromatgraphy (silica, 2:1 O^C^/acetone) gavē 106 as a pale yellow solid. TLC Rf = 0.19 (2:1 CH2Cl2/acetone): lE NMR (CDCI3. 400 MHz) δ 7.30 (m, 3H), 6.23 (t, J=7 Hz, 1H), 4.04 (m, 2H), 2.69 (m, 2H), 2.57 (m, 2H), 1.74 (m, 2H), 1.56 (m, 2H), 1.48 (m, 1H), 1.47 (s, 9H), 1.16 (m, 2H). - 112 -

Preparation of Ethyl {3-[2-(N-B0C-piperidin-4-yl)- ethvl1-2-pvr idon-l-vllacetate._ 1107)_

To a stirred solution of 106 (0.44 g, 1.4 mmol) in dry DMF (7.2 mL) at 0eC was added NaN(TMS)2 (2.1 mL, 2.1 mmol, lM/hexanes) dropvise. After 30 min. ethyl bromoacetate (0.79 mL, 7.2 mmol) was added dropwise to the reaction mixture. After an additional hour the reaction was diluted with ethyl acetate and then washed with H20, 5% KHSO4, sat. NaHC03 and brine, dried (MgSO^) and concentrated. Flash chromatography (silica, 457. ethyl acetate/hexanes) gavē 107 as a colorless oil. TLC Rf = 0.33 (607. ethyl acetate/hexanes); 7.20 (dd, J=6 and 1 Hz, 1H), 7.11 (dd, J=7 and 2 Hz, 1H), 6.15 (t, J=7 Hz, 1H), 4.62 (s, 2H), 4.24 (q, J=7 Hz, 2H), 4.04 (m, 2H), 2.69 (m, 2H), 2.55 (m, 2H), 1.72 (m, 2H), 1.52 (m, 2H), 1.78 (τη, 1H), 1.48 (s, 9H), 1.13 (m, 2H).

Preparation of {3-[2-(N-B0C-piperidin-4-yl)ethyl]-2- pvridon-l-vI&gt; acetio acid (108)_

Utilizing the procedure for converting 5i to 56. 107 (0.51 g, 1.3 mmol) gavē 108 as a colorless foam. TLC RF = 0.58 (9:1:1 CH2C12/CH30H/H0Ac). - 113 - LV10425

Preparation of {3-[2-(N-B0C-piperidin-4-yl)ethyl]-2-pyridon-l-yl}acetyl-3(R)-methyl-B-alanine ethyl ester asm_:_

Utilizing the procedue for converting 15. to 16. 108 (0.15 g, 0.41 mmol) gavē 109 as a waxy solid after flash chromatography (silica, ethyl acetate). TLC Rf = 0.38 (ethyl acetate) XH NMR (CDC13, 400 MHz) δ 7.24 (dd, J=7 and 2Hz, 1H), 7.21 (dd, J=7 and 2 Hz, 1H), 7.07 (bd, J=8 Hz, 1H), 6.19 (t, J=7Hz, 1H), 4.51 (s, 2H), 4.30 (m, 1H), 4.08 (q, J=7Hz, 2H), 4.08 (m, 2H), 2.69 (m, 2H), 2.55 (m, 2H), 1.72 (m, 2H), 1.51 (m, 2H), 1.45 (s, 9H), 1.43 (m, 1H), 1.23 (t, J=7Hz, 3H), 1.22 (d, J*7Hz, 3H), 1.13 (m, 2H).

Preparation of {3-[2-(N-B0C-piperidin-4-yl)ethyl]-2- pYridon-l-yl)acetvl-3(R)-methyl-J3-alanine .(110)-

Utilizing the procedure for converting 5i. to 56. 109 (0.19 g, 0.41 mmol) gavē 110 as a colorless oil. TLC = 0.59 (9.5/0.5/0.5 CH2C12/CH3OH/HOAc).

Preparation of {3-[2-(Piperidin-4-yl)ethyl]-2-pyridon- l-vl)acetvl-3(R)-methvl-B-alanine (111)_ A solution of 103 (0.18 g, 0.40 mmol), CH2C12 (2.0 mL), anisole (86 μL, 0.80 mmol), and trifluoroacetic acid (2.0 mL) was stirred at -15* for 15 min. The yellow reaction mixture was concentrated and the residual trifluoroacetic acid removed azeotropically with toluene. Flash chromatography (silica, 10/1.3/1.3 ethanol/NH40H/H20) give 1LL as a white amorphous solid. 114 - TLC Rf = 0.17 (10:1:1 ethanol/NH40H/H20); ΧΗ NMR (D20, 400 MHz) δ 7.57 &lt;dd, J=7 and 1 Hz, 1E), 7.49 (dd. J=7 and 1Hz, 1H), 6.51 (t, J=7Hz, 1H), 4.66 (m, 2H), 4.20 (m, 1E), 3.42 (m, 2H), 2.97 (m, 2H), 2.59 (m, 2H), 5 2.46 (dd, j=14 and 6 Hz, 1H), 2.32 (dd, J=14 and 7Hz, 1H), 2.00 (m, 2H), 1.60 (m, 3H), 1.42 (m, 2H).

Prenaration of N-Cbz-4-pjperidiaefithanoL-(112)

To a stirred solution of 4-piperidineethanol 10 8 (15 g, 0.12 mol), THF (500 mL), and diisopropylethyl amine (40.5 mL, 0.23 mol) at 0eC was added benzyl chloroformate (16.5 mL, 0.12 mmol). After 1 h at 0*C the reaction mixture was diluted with ethyl acetate and then washed with H20 (2x), 107. KHS04, and brine, 15 dried (MgS04), and concentrated to give 112 as a colorless oil. TLC Rf = 0.60 (ethyl acetate); % NMR (300 MHz, CDC13) δ 7.35 (m, 5H), 5.13 (s, 2H), 4.18 (m, 2H), 3.70 (&lt;j, 20 J=7Hz, 2H), 2.80 (m, 2H), 1.80-1.50 (m, 5H), 1.19 (m, 2H).

Preparation of N-Cbz-4-piperidine ethvl iodide (113^ Utilizing the procedure for converting 3. to 25 112 (30.6 g, 0.12 mol) gavē 113 as a colorless oil after flash chromatography (silica, 107. ethyl acetate/hexanes). 30 - 115 - LV10425

Preparation of l-[2-(N-Cbz-piperidin-4-yl)ethyl]- (2-pvrrolidinone)_· (114).__

To a stirred solution of 2-pyrolidinone (11.0 g, 0.13 mol) in dry DMF (500 mL) at ambient temperature was added NaN(TMS&gt;2 (129 mL, 129 mmol, 1M/THF) dropwise. After 10 min, the iodide 113 (24.1 g, 64.6 mmol) in DMF (50 mL) was added to the reaction vessel. After 1 hour, the reaction mixture was diluted with ethyl acetate and then washed with H2O, sat. NaHC03 and brine, dried (MgSC&gt;4) and concentrated. Flash chromatography (silica, ethyl acetate) gavē 114 as a colorless oil. TLC Rf=0.16 (ethyl acetate); NMR (300 MHz, CDCI3) δ 7.40-7.20 (m, 5H), 5.12 (s, 2H), 4.19 (m, 2H), 3.34 (m, 2H), 2.77 (m, 2H), 2.40 (m, 2H), 2.04 (m, 2H), 1.74 (m, 2H), 1.47 (m, 3H), 1.15 (m, 2H).

Preparation of l-[2-(N-Cbz-piperidin-4-yl)ethyl]-3-propen-2-vl-(2-pvrrolidinone) (115_).-—

To a stirred solution of 114 (8.0 g, 24.2-mmol), and THF (180 mL) at -78°C was added LDA (50 mL, 25 mmol, 0.5M/THF) dropwise. After 15 min. allyl 1· bromide (2.3 mL, 26.6 mmol) was added. After 1.0 hour, the reaction mixture was allowed to warm to 0eC over a 10 min. period. The reaction mixture was diluted with ethyl acetate and then washed with 57. KHSO4 and brine, dried (MgS04&gt; and concentrated.

Flash chromatography (silica, 507. ethyl acetate/ hexanes) gavē 115. - 116 - TLC Rf=0.65 (ethyl acetate); ^-H NMR (300 MHz, CDC13) 6 7.35 (m, 5H), 5.80 (m, 1H), 5.12 (m, 2H), 4.18 &lt;m, 2H), 3.33 (m, 4H), 2.80 (m, 2H), 2.56 (m, 2H), 2.20 (m, 2H), 1.78 (m, 2H), 1.48 (m, 3H) , 1.20 (m, 2H).

Preparation of l-[2-(N-Cbz-piperidin-4-yl)-ethylJ-3-acetic acid-2-Pvrrolidinone (116)._

Utilizing the procedure for converting 14 to 15. 115 (4.7 g, 12.7 mmol) gavē 116 as a light brown oil after flash chromatography (silica, CH2CI2/CH3OH/ HOAc 9.5:0.5:0.5). TLC Rf=0.63 (CH2C12/CH30H/H0Ac 9.0:0.5:0.5); 1H-NMR (300 MHz, CDCI3) 5 7.37 (m, 5H), 5.15 (s, 2H), 4.18 (m, 2H), 3.37 (m, 4H), 3.00-2.30 (m, 5H), 1.77 (m, 2H), 1.46 (m, 3H), 1.18 (m, 2H).

Preparation of [l-[2-(N-Cbz-piperidin-4-yl)ethyl]-2-pyrrolidinone-3]acetyl-2-aminoethylphosphonic acid (117)._:_1 A solution of 116 (388 mg, 1.0 mmol), Et3N (0.14 mL, 1.0 mmol), and dry dioxane (5 mL) at 0*C was treated with isobutyl chloroformate (0.13 mL, 1.0 mmol). The reaction mixture was warmed until the dioxane melted. A solution of 2-aminoethylphosphonic acid (125 mg, 1.0 mmol), Na2CŪ3 (106 mg) and H20 (2 mL) was added to the reaction vessel at ambient temperature. After 4 hours the dioxane was evaporated and the residue acidified with 1N HC1. The mixture - 117 - LV10425 was washed with ethyl acetate &lt;3x). The aqueous portion was concentrated to dryness to afford crude UZ. TLC Rf=0.15 (9:1.0:1.0 CH2Cl2/CH30H/H0Ac).

Preparation of [l-[2-(piperidin-4-yl)ethyl]-2-pyrrolidinone-3]acetyl-2-aminoethylphosphonic acid (118),._

The crude phosphonic acid 117 (464 mg) was dissolved in 10% HOAC/CH3OH (20 mL), then Pd(0H&gt;2 (50 mg) was added and the mixture hydrogenated on the Parr apparatus (50 PSI) overnight. After 20 hours the reaction mixture was filtered through a celite pad and the filtrate concentrated. Flash chromatography (silica, 8-r-l:l CH30H/H20/NH40H) gavē 118 as an amorphous solid. TLC Rf=0.14 (8:1:1 CH30H/H20/NH40H); ΧΗ NMR (300 MHz, H20) δ 3.40 (m, 8H), 3.18 (m. 1H), 2.92 (m, 3H), 2.60 (m, 1H), 2.38 (m, 1H), 2.26 (m, 1H), 1.97 (m, 2H), 1.79 (m, 3H), 1.56 (m, 2H), 1.40 (m, 2H).

Preparation of (4-N-Cbz-amino)cyclohexane carboxylic acid _m£t.hyl ester ..(1.21)..,__

To a suspension of 120 (17.5 g, 0.12 mol) in CH3OH (150 mL) at -10*C was added thionyl chloride (13.4 mL, 0.18 mol) portionwise over a 5 min. period, followed by removal of the cooling bath. After 16 hours the resulting solution was concentrated and the residue dissolved in DMF (150 mL), cooled to 0eC then treated sequentially with N(i-Pr)2Et (52 mL, 0.3 mol) and benzyl chloroformate (18.6 mL, 0.13 mol). The - 118 - cooling bath was removed and after 24 hours the reaction mixture was concentrated. The residue was diluted with ethyl acetate and then washed with saturated NaHCOg, 5% KHSO^. and brine, dried (MgSO/ļ) and concentrated. Flash chromotography (silica, 20% ethyl acetate/hexanes) gavē 121 as a waxy crystalline solid. TLC Rf=0.34, 0.39 (30% ethyl acetate/ hexanes); XH NMR (400 MHz, CDC13) δ 7.38 (m, 5H), 5.09 (bs, 2H), 4.68 (m, 0.5H), 4.59 (m, 0.5H), 3.72 (m, 0.5H), 3.68 (s, 1.5H), 3.67 (s, 1.5H), 3.50 (m, 0.5H) 2.45 (m, 1H), 2.13-1.50 (m, 8H).

Preparation of (4-N-Cbz-amino)[l-(propen-3-yl)]cyclo-hexane carboxvlic acid methvl ester (122)._

To a stirred solution of 121 (1.0 g, 3.4 mmol) in dry THF (17.2 mL) at -78eC was added LDA (4.3 mL, 8.6 mmol, 2.0M) dropwise. The cooling bath was warmed to -40eC. After stirring at -40eC for 15 min. the reaction mixture was recooled to -78eC and then treated sequentially with 1,3-dimethyl-3,4,5,6-tet-ra-hydro-2-(lH-pyrimidinone) (2.0 mL, 17 mmol) and allyl bromide (0.44 mL, 5.1 mmol). After 2 hours the reaction was diluted with ethyl acetate and then washed with Η£θ), sat. NH^Cl, sat. NaHCOj and brine. Drying (MgSO^) and concentration gavē a yellow oil which was subjected to flash chromatography (silica, 207. ethyl acetate/hexanes) to give 122 as a waxy crystalline solid. TLC Rf=0.31 (20% ethyl acetate/hexanes); NMR (400 MHz, CDCI3) 5 7.34 (m, 5H), 5.67 (m, 1H), 5.10 (s, 2H), 5.05 (m, 2H), 4.54 (m, 1H), 3.67 (s, 3H), 3.48 (m, 1H), 2.21 (m, 2H), 2.00-1.00 (m, 8H). - 119 - LV10425

Preparation of 4-Amino,l-(propen-3-yl)cyclohexane carboxvlic acid methvl ester (123)_

Trifluoroacetic acid (19 mL) was added to a mixture of 122. (0.63 g, 1.9 mmol) and anisole (0.41 ml, 3.8 mmol) at 0*C. After 5 min. the reaction mixture became homogeneous and removal of the cooling bath was followed by stirring overnight.

Concentration, folloved by flash chromatography (silica, 17:1:1 ethanol/NH^OH/^O) gavē 121 as .a yellow oil. TLC Rf=0.53 (30% ethyl acetate/hexanes).

Preparation of [2-Aza-3-oxo-[2.2.2]bicyclooct-4-yl] prop-3-ene (124)._ A solution of 123 (0.39g, 1.97 mmol) in toluene (10 mL) was heated at 170eC in a sealed tube overnight. The cooled dark reaction mixture was purified by flash chromatography (silica, 85% ethyl acetate/hexanes) to give 124 as a tan oil. TLC Rf=0.30 (85% ethyl acetate/hexanes); l-HMNR (400 MHz, CDCI3) 5 6.08 (m, 1H), 5.88 (m, 1H), 5.04 (m, 2H), 3.61 (m, 1H), 2.35 (d, J=7Hz, 2H), 1.80-1.50 (m, 8H).

Preparation of ([2-(N-Boc-piperidin-4-yl)ethyl]-2-aza-3-oxo-r2.2.21bicvclooct-4-vl)prop-3-ene (125).

Utilizing the procedure for converting 36a to IZa, 12A (0.18 g, 1.1 mmol) gavē 125. (0.45 g, 100%) after flash chromatography (silica, 40% ethyl acetate/ hexanes). 120 TLC Rf=0.36 (407. ethyl acetate/hexanes); ^-H NMR &lt;400 MHz, CDCI3) δ 5.87 (m, 1H), 5.05 &lt;m, 2H), 4.08 (m, 2H), 3.51 (m, 1H), 3.40 (t, J=8Hz, 2H), 2.67 (m, 2H), 2.34 (d, J=7Hz, 2H), 1.75-1.35 (m, 13H), 1.47 (s, 9H), 1.12 &lt;m, 2H).

Preparation of 3-(l-[2-(N-Boc-piperidin-4-yl)ethyl]-2-β·ζα-1-οχο-Γ2.2.2~lbicvclooct-4-vl)acetic acid (126)._

Utilizing the procedure for converting 14 to 15. 125 (0.42 g, 1.1 mmol) gavē 126 (0.34 g, 76%) as a white sticky foam.. TLC Rf=0.58 (9.5:0.5:0.5 CH2C12/CH3OH/AcOH).

Preparation of ([2-(N-Boc-piperidin-4-yl)ethyl]-2-aza-3-oxo-[2.2. 2]bicyclooct-4-yl)acetyl=-B-alanine tert-butvl ester (127)._

Utilizing the procedure for converting 15. to li, 12A (0.14g, 0.35 mmol) gavē 127 (0.10 g, 55%) as a colorless oil after flash chromatography (silica, 6:1 CH2Cl2/acetone). TLC Rf=0.52 (2:1 CH2Cl2/acetone); ΧΗ NMR-(400 MHz, CDCI3) δ 7.90 (m, 1H), 4.04 (m, 2H), 3.56 (m, 1H), 3.42 (m, 4H), 2.67 (m, 2H), 2.42 (t, J=7Hz, 2H), 1.76-1.34 (m, 13H), 1.46 (s, 18H), 1.12 (m, 2H).

Preparation of ([2-(Piperidin-4-yl)ethyl]-2-aza-3-oxo-r2.-l.-21bicvr-loopt-4-vl)acetvl-B-alanine (128)._.

Utilizing the procedure for converting 40a to 4.1a, 127 (0.10 g, 0.19 mmol) gavē 128 (29 mg, 39%) as a fluffy solid after flash chromotography (silica, 10:1.2:1.2 ethanol/H20/NH40H). - 121 - LV10425 TLC Rf=0.17 (10:1:1 ethanol/NH40H/H20); ^ NMR (400 MHz, D20) δ 3.78 (m, 1H), 3.50-3.35 (m, 6H), 2.97 (m, 2H), 2.48 (s, 2H), 2.38 (t, J=7Hz'‘, 2H), 2.02 (m, 2H), 1.78 (m, 6H), 1.58 (m, 5H), 1.42 (m, 2H).

Applicants hereby incorporate by reference procedures for preparing compounds of the present invention whereby guanidines are prepared from amines and whereby amidines are prepared from corresponding nitriles. Guanidines may be prepared from amines by those having ordinary skill in the art upon reaction with 3,5-dimethylpyrazole-l-carboxamidine nitrate (Methods Enzvmol.. 25b, 558, 1972). Amidines may be prepared from the corresponding nitrile by those having ordinary skill in the art using procedures _ demonstrated by Boere, R.T., et. al. J. Organomet. Chem..., 331(2), 161-7, 1987; and Fuks, R. , Tetrahedron. 29 (14), 2147-51, 1973.

Utilizing the methodology demonstrated in this invention, the following compounds, included in Table I below, are exemplary of the compounds which may be prepared according to this invention. 122 -

TAELE I R1-CH2-C-NH-R2 o H2N-CH2 'ό''' -ch-ch2co2h

NH o .N-^NH-C CH2) 3-N'^S1' -ch-ch2co2h

N. &gt;=o s N^-CC^)3-ļ^N-

NH

II

HjN-C-CC^)*- -ch2-ch-co2h -ch-ch-co2h

- 123 -LV10425 TABLE I (Cont1d)

R h2n-ch3— J2 — «2 ‘ o R3 CH3Ph o-. 1 &quot;C-CH-C03 I Λ -ch-ch3co; N N— CH, 10

HN

O

CH-CHj-COjH O' —ch3 ch3 20

HN •N- ch3-ch3-co3h o

II C-Pha

-CH-CH3-C03H

O NH 25 H2N-C-CCH3)3_^ o II C-Pha-Lau -ch-ch3 co3h 30 124 -

In vivo Activitv

The following protocols were used to determine in vivo activity in dogs of fibrinogen receptor antagonists of the present invention. A mongrel dog of either sex (weighing 7.5 to 11.5 kg) is comfortably positioned in a nylon sling which exposes the legs and keeps the dog stationary. Five ml of blood is withdrawn from either saphenous or cephalic veins with a plastic syringe containing 0.5 ml of 3.8% citrate via a 19-gauge butterfly. An additional 1 ml of citrated blood is taken to measure whole blood platelet counts. Ex vivo platelet aggregations are done with the agonists ADP (10 μϋ) and collagen (10 μg/ml) both primed with 1 μΜ epinephrine. Platelet-rich plasma (PRP) is obtained by centrifuging the blood at 150 x g for 5 minūtes. Platelet counts are adjusted to 200,000/mm·^ with platelet-poor plasma.

Oral administration of a drug is done either as a gelatin capsule or a gastric lavage. 'Por the-gastric lavage method, 5 ml of the drug solution is administered to the dog through a feeding tube.

Blood samples for ex vivo platelet aggregation are taken at the folloving time points: 0,20, 40, 70, 90, 150, 200, 250, 300, 350, 480 min. 24 hr. 30 hr, and 48 hr. At each time point, the remaining platelet-poor plasma is saved and frozen for drug Ievels. Three additional blood samples are taken at 30, 55, and 110 minūtes after dosing for drug plasma Ievels. LV10425 - 125 -

For the intravenous infusion, blood samples for ex vivo platelet aggregation are taken at the folloving time points: 0,30, 45, 60, 90 and 120 min into the infusion and 2,5, 15, 30, 60, 90, 120, 180, 240, 300, 360, and 420 minūtes after the infusion stops. At each time point, the remaining platelet-poor plasma is saved and frozen for drug Ievels. The drug is infused for 120 minūtes at a constant rāte of 0.1 ml/min.

For the intravenous bolus, 5 ml of the drug solution is administered quickly. Blood samples for ex vivo platelet aggregation are taken at 0, 1, 5, 10, 15, 30, 45, 60, 90, 120, 180, and 240 minūtes after the bolus. At ali time points, the remaining platelet-poor plasma was saved and frozen for drug I.

Ievels.

In the oral and intravenous groups, blood collection times are recorded at each time point in order to determine drug plasma Ievels.

The folloving compound

HN

O O

H was evaluated according to the procedures described above. - 126 -

Inhibition of platelet aggregation of control) induced by intravenous infusion at rātes of 10 μg/kg/min and 25 μg/kg/min was measured. Inhibition was nearly complete during infusion and partial inhibition existed for several hours after infusion cessation.

Inhibition of platelet aggregation (7. of control) induced by single oral administration of 2 rag/kg was also measured. % Inhibition of platelet aggregation was greater than 507. for a period of eight hours. - 127 - 10 15 20 25 WHAT IS CLAIMED IS: LV10425 1. .A fibrinogen receptor antagonist of

Formula I: vherein: X is

0 R II i X-(CH2)m-Y-(CH2)k-C-NH-CH-CH-Z NH NH NH -NR7R3, -nh-c-nr7r3, -c-nr7r3, r7-c-nh-, h2n-ch2 0 where D= , -S(0)q, or -0-; 30 128 5

NH R4 7 'Ή R7-N 1 N

.4 10 15 where A=N and B= -CH2-. or A- CH and B-N-R7; Y is

30 - 129 - LV10425 Ο 0 Z is -COoR2; -P-0H -P-OH OR2 ; R10 ; where R10 is Cļ_galkyl, aryl, arylC1_8alk.yl u is -CH-, -C-, or -N-; v is -CH-, -C-, or -N-; R and R·*· are independently hydrogen, aryl, wherein aryl is defined as a mono- or polycyclic aromatic system comprised of 5 or 6 membered rings containing 0, 1, 2, 3, or 4 heteroatoms selected from nitrogen, oxygen and sulfur; and phenylene, either unsubstituted or substituted, with one or more groups selected from hydroxyl, fluoro, chloro, bromo, iodo, cyano, trifluoromethyl, Cļ_3 alkoxy, Cļ_5alkylcarbonyloxy, C]__5alkoxycarbonyl, C]__5alkyl, aminoC]__5alkyl, hydroxycarbonyl-CQ_5alkyl, or hydroxycarbonylCļ_5alkoxy, CQ_6alkyl, either unsubstituted or substituted, with one or more 130 groups selected from fluoro, chloro, bromo, iodo, hydroxyl, Cļ_5alkyl-carbonyl(CQ_Qalkyl)amino, arylCļ_5alkyl carbonyl(CQ_galkyl)amino, aryloxy, Cļ_iQ alkoxy, Cļ_5alkoxycarbonyl, Co_5alkylaminocarbonyl, Cļ_5alkyl-carbonyloxy, Cg_g cycloalkyl, aryl, oxo, amino, Cļ_g alkyl, Cļ_3alkylamino, aminoCļ_3 alkyl, arylCQ_5alkylamino-carbonyl, phenylC]__3alkylami.no, amino-carbonyl-CQ_4alkyl, Cļ_8alkylsulfonyl (Co_8alkyl)amin0, aryl Co-lO3·11^1&quot; sulfonyl(CQ_galkyl)amino,arylCQ_galkyl-sulfonyl, CQ_galkylsulfonyl, hydroxy-carbonylCo_5 alkyl, Cļ_galky-loxycarbonyl (Co_8alkyl)amino, arylCQ_ļ0alkyloxy-carbonyl(CQ_galkyl)amino» Cg_galkyl-aminocarbonyl(Co_8 alkyl) amino, arylCQ_galkylaminocarbonyl(Co_8alky1)- amino, CQ_galkylaminocarbonyloxy, arylCQ_10alkylaminocarbonyloxy, CQ_galkylaminosulfonyl(CQ_g alkyl)-amino, arylCQ_galkylaminosulfonyl(CQ_g-alkyl)amino, CQ_galkylaminosulfonyl, or arylCg_g-alkylaminosulfonyl; provided that the carbon atom to which R or R^ is attached bear only one heteroatom; - 131 - LV10425 R2 is hydrogen,

Ci_i2alkyl, unsubstituted or substituted, with one or more Cļ_^alkyl groups, 0 R9 0 CH? CH-, 0

&quot; Q 1 11 ο J ii Q -CH20CR9, -CH0CR9 or -C-0-C-R9 where R9 = Cļ_6alkyl, branched or unbranched, or phenyl, and wherein R9, when appearing more than once, can be the same or different; R7, R3 and R^ are independently hydrogen, cl-12 alky1» unsubstituted or substituted, with one or more Cļ_^ alkyl groups, or arylCQ_4alkyl, or cyano, provided that when R7 and R3 are independently cyano,

NH NH X is -NH-C-NR7R3 or -C-NR7-R3; k is 1-4; m is 1-4; p is 1-6; q is 0-2; salts thereof or or the pharmaceutically acceptable optical isomer thereof. 132 - 2. A fibrinogen receptor antagonist of formula 1 in Claim 1, vherein:

NH X is -NR7R3, -C-NR7R3, CCH2)p

0 u where D = -C-, -S(0)q, or -0-; Y is LV10425

where n is 1, 2 or 3, Z is CC^R^ R and R^· are independently chosen from phenyl, thiophene, imidazole, naphthyl, indole, indazole, thionaphthene, either unsubstituted or substituted, with hydroxy, halogen, hydroxycarbonylCQ_5 alkyl, Cj__3alkyl, either unsubstituted or substituted, with one or more groups selected form aryl, aryloxy, Cļ_ļQ alkoxy, CQ_5alkylaminocarbonyl, arylCQ_5 alkylaminocarbonyl, hydrogen, 134 -

Cg_galkyl either unsubstituted or substituted, with one or more groups selected from halogen, hydroxyl, Cļ_galkylsulfonylamino, aryl Cg_^alkylsulfonylamino, Cļ_^alkylsulfonyl, aryl Cg_^alkylsulfonyl, Cļ_5alkylcarbonylamino, arylC1_5alkylcarbonylami.no, aryloxy, Cļ.ļg alkoxy, Cļ_5alkoxycarbonyl, CQ_5alkylamino-carbonyl, Cļ_5alkylcarbonyloxy, C3_gcycloalkyl, aryl, 0x0, amino, Ci_5alkyl, Ci_3alkylamino, aminoCi_3alkyl, arylCo_5alkylaminocarbonyl, phenylC1_3alkylami.no, aminocarbonylCQ_4alkyl, or hydroxycarbonylCg_5alkyl, provided that the carbon atom to which R or R* is attached bear only one heteroatom, is hydrogen,

Ci_i2alkyl» unsubstituted or substituted, with one or more Cļ_galkyl groups, 0 R9 0 CH3 CH3 0 -CH20CR9, -CH0CR9 or -C-0--C-R9 where R9 = Cļ_galkyl, branched or unbranched, or phenyl; R^, R^ and R^ are independently hydrogen, Cļ_3alkyl, unsubstituted or substituted, with one or more Cļ_galkyl groups; k is 1-4; m is 1-4; P is 1-3; is 0 or 2; or the pharmaceutically acceptable salts thereof, or optical isomer thereof. - 135 - LV10425 3. A fibrinogen receptor antagonist of Formula I in Claim 1, wherein X is R4 R7-n°^r&lt; CCH2)p R’-N^N -NR7R3.

&quot;V H. , or h2n-ch2 0 u where D = -0-, -S-, or -C-Z is C02R2 Y is

wherein n is 1, 2 or 3; 136 - R and are independently chosen from phenyl, thiophene, iraidazole, naphthyl, indole, indazole, thionaphthene, either unsubstituted or substituted, with hydroxy, halogen, hydroxy-carbonylCQ_5alkyl, Cļ_3alkyl, either unsubstituted or substituted, with one or more groups selected form aryl, aryloxy, Cļ_ļQ alkoxy, CQ_5alkylaminocarbonyl, arylCg_5 alkylaminocarbonyl, hydrogen, CQ_galkyl, either unsubstituted or substituted,with one or more groups selected from halogen, hydroxyl, Cļ_5alkylcarbonylamino, aryl Cļ_5alkylcarbonylamino, aryloxy, C2_2Qalkoxy, Cļ_5alkoxycarbonyl, CQ_5alkylaminocarbonyl, Cļ_5alkylcarbonyloxy, C3_gcycloalkyl, aryl, 0X0, amino, Cļ_galkyl, Cļ_3alkylamino, amino Cļ_3 alkyl, arylCQ_5alkylaminocarbonyl, phenylC2_3alkylamino, aminocarbonylCQ_4 alkyl, or hydroxycarbonylC0_5alkyl, provided that the carbon atom to which R or R^- is attached bear only one heteroatom; R2 is, hydrogen; R7, R2 and R^ are hydrogen; k is 1-2; m is 1-4; p is 2-4; or the pharmaceutically acceptable salts thereof, or optical isomer thereof. - 137 - LV10425 4. The compound of Cļaim 1 for use in inhibiting the binding of fibrinogen to blood platelets, inhibiting the aggregation of blood platelets, treating thrombus formation or embolus formation, or preventing thrombus or embolus formation in a mammai. 5. A compound of Claim 1 selected from the group consisting of:

O H 138 5

ο2η 10

25

.^^COOH 30 - 139 - LV10425

30 140 CHq Γ^Ι,Η Ο Νν ΗΝ

ΝΗ A^COOH Ο

ΗΝ Ο ^cooh Η

- 141 -LV10425

HN

ΌΗ

142

10

15

25

30 - 143 - Ν·

HN o Ο x\^po3 h2 ,

H LV10425

or pharmaceutically acceptable salts thereof. 6. A compound of claim 5 which is: SCH3

or pharmaceutically acceptable salts thereof. 144 - 7. A compound of claim 5 which is: 144 -

or pharmaceutically acceptable salts thereof. 8. A compound of Claim 5 which is:

o2h or pharmaceutically acceptable salts thereof. 9. A compound of Claim 5 which is:

or pharmaceutically acceptable salts thereof. - 145 - LV10425

10 or pharmaceutically acceptable salts thereof 11. A compound of Claim 1 which is 15 *

H

20 25 30 146 Ο Ο

or pharmaceutically acceptable salts thereof. 12. A composition for inhibiting the binding of fibrinogen to blood platelets in a mammai, comprising a compound of Claim 1 and a pharmaceutically acceptable carrier. 13. A composition for inhibiting the aggregation of blood platelets in a mammai, comprising a compound of Claim 1 and a pharmaceutically acceptable carrier. 14. A composition for inhibiting the aggregation of blood platelets in a mammai, comprising a compound of Claim 1 in combination with a thrombolytic aģent and a pharmaceutically acceptable carrier. - 147 - LV10425 15. A composition for inhibiting the aggregation of blood platelets in a mammai, comprising a compound of Claim 1 in combination with an anticoagulant and pharmaceutically acceptable carrier. 16. A composition for preventing and/or treating thrombus or embolus formation in a mammai, comprising a compound of Claim 1 and a pharmaceutically acceptable carrier. 17. A composition for preventing and/or treating thrombus or embolus formation in a mammai, comprising a compound of Claim 1 in combination with a thrombolytic aģent and a pharmaceutically acceptable carrier. 18. The composition of Claim 14 or Claim 17, wherein the thrombolytic aģent is a plasminogen activator or streptokinase. 19. A composition for preventing and/or treating thrombus or embolus formation in a mammai, comprising a compound of Claim 1 in combination with an anticoagulant and a pharmaceutically acceptable carrier. 20. The composition of Claim 15 or Claim 19, wherein the anticoagulant is heparin or varfarin. 148 21. A composition for treating thrombus or embolus formation in a mammai, comprising a compound of Claim 1 in combination with an antiplatelet aģent and a pharmaceutically acceptable carrier. 22. The composition of Claim 1, vherein the antiplatelet aģent is aspirin. 23. The use of a composition of Claim 12 for the manufacture of a medicament for inhibiting the binding of fibrinogen to blood platelets in a mammai. 24. The use of a composition of Claim 13, 14 or 15 for the manufacture of a medicament for inhibiting the aggregation of blood platelets in a mammai7~ 25. The use of a composition of Claim 16, 17 or 19 for the manufacture of a medicament for preventing and/or treating thrombus or embolus formation in a mammai. 26. The use of a composition of Claim 21 for the manufacture of a medicament for treating thrombus or embolus formation in a mammai.

Claims (26)

LV10425 FORMULA OF THE INVENTION 1. A fibrinogen receptor antagonist of the formula: O R R 'where x is NH NH NH li li I! -NR7R1-NH-C-NH'R1, -C-NR7R1'-c-nh-, o II wherein D = -C-, -S (O) q or -O-; wherein A = N and B = -CH 2 -, or A = -CH- and B = N-R 4 -; Y and 2 R ' Ο 3 Ο Ο II II ζ and -CO.R1; -Ρ-ΟΗ; -Ρ-ΟΗ ;, I I OR 1 R '° where C 1 -C 17 alkyl, aryl, aryl C 1 -C 2 alkyl; u is -CH-, -C- or -N-; v is -CH-, -C- or -N-; RunR1 is independently from each other hydrogen, aryl, where aryl is defined as a mono- or polycyclic aromatic system with 5- or 6-membered rings containing 0, 1, 2, 3 or 4 heteroatoms which may be nitrogen, oxygen and sulfur; and phenylene, unsubstituted or substituted with one or more groups which may be hydroxy, fluorine, chlorine, bromine, iodine, cyano, trifluoromethyl C 1-3 alkoxy, C 1-5 alkylcarbonyloxy, C 1-5 alkoxycarbonyl, C 1-5 alkyl, amino-C 1-5 alkyl, hydroxycarbonyl-C 0-5 alkyl, or hydroxycarbonyl-C 1-5 alkoxy; C 1-6 alkyl, unsubstituted or substituted with one or more groups which may be fluoro, chloro, bromo, iodo, hydroxy, C 1-5 alkylcarbonyl (O-galkyl) amino, aryl-C 1-5 alkylcarbonyl (Co- 8a * k'0 amino-, aryloxy-, C 1-4 alkoxy-, C 1-5 alkoxycarbonyl-, C 1-6 alkylaminocarbonyl-, C 1-6 alkyloxycarbonyloxy-, ¢ 3.3 cycloalkyl-, aryl-, oxo-, amino-, alkyl, C 1-3 alkylamino, amino-C 1-3 alkyl, aryl-C 1-8 alkylaminocarbonyl, phenyl-C 1-6 alkylamino, aminocarbonyl-C 1-4 alkyl, C 1-4 alkylsulfonyl (Co.8 alkylamino, aryl-C8-alkyl, kilosulfonyl (C0-C8 alkyl) amino, aryl-C0-alkylsulphonyl, C8-alkylsulphonyl, hydroxycarbonyl-3-alkyl, C-5 alkyl). e.alkyloxycarbonyl (Co-8a, Ki) amino-C0-6alkylaminocarbonyl (C0-4alkyl) amino, aryl-C0-8 alkylaminocarbonyl (C0-8alkyl) amino, C0-8alkylaminocarbonyloxy, or aryl-C0-10alkyl ' nicarbonyl) xi &gt; Co-8alkylaminosulfonyl (Co-s) alkylamino, Co-8a, kil * aminosulfonyl or aryl-C0alkylaminodulfonyl provided that the carbon to which the R and R 1 groups are attached is bound to only one heteroatom; R 2 is hydrogen, C 1 -C 12 alkyl unsubstituted or substituted with one or more C 1 -C 6 alkyl groups, O R 'O CH 3 CH 3 O II I II ✓ -CH 2 OCR', -CHO-CR * or -COC-R 'where R 9 = C 1-6. galyl, branched or unbranched, or phenyl, wherein if the R 9 groups appear more than once in the formula, they may be the same or different. R 7, R 3 and R * are independently from each other hydrogen, C 1 -C 12a, unsubstituted or substituted with one or more C 1 -C 4 alkyl groups, or aryl-C 0 -C 4 alkyl, or cyano groups, wherein R 7 and R 3, independently of one another being cyano, then NH NH II II X is -NH-C-NR 7 R J or -C-NR 7 R 4 k is 1 * 4; m is 1 to 4; p is 1-6; q is 0-2; or pharmaceutically acceptable salts thereof, or optical isomers thereof. Fibrinogen receptor antagonists of formula I according to claim 1, wherein NH II X is -NR7R \ -C-NR7R \ 4 Ο II wherein D = -C-, -S (O) q or -O-; Y and R or R Wherein n is 1, 2 or 3; z is CO2R2 R and R1 are independently selected from the group consisting of phenyl, thiophene, imidazole, naphthyl, indole, indazole, 2,3-benzothiophene, unsubstituted or substituted with one or more groups which may be aryl, aryloxy, C 1-6 alkyloxy, C 0 -C 5 alkylalkenylcarbonyl, arylC 0-5 alkylaminocarbonyl, hydrogen, C 8 -galkyl, unsubstituted or substituted with one or more groups which may be halogen, hydroxy, C 1-8 alkyl -, C 1-6 alkylsulfonyl, arylC 0-6 alkylsulfonyl, C 1-6 alkylcarbonylamino, aryl-C 1-5 alkylcarbonylamino, aryloxy, C 1-3 galcoxy, C 1-6 alkoxycarbonyl.Cg. galkylaminocarbonyl, C 1-8 alkylcarbonyloxy, C 3-8 cycloalkyl, aryl, oxo, amino, C 1-8 alkylalkyl, C 1-3 alkylamino, amino-C 1-3 alkyl, aryl-Co- salkylaminocarbonyl, fertyl-C 1-6 alkylamino, aminocarbonyl-C 0-4 alkyl or hydroxycarbonyl-C 8 -alkyl, provided that the carbon atom to which R and R 1 are attached is bonded to only one heteroatom, R 2 is hydrogen, C 1-6 alkyl, unsubstituted or substituted with one or more C 1-4 alkyl, O R 'O CH 3 CH 3 O II I II ✓ -CH 2 OCR 1, -CHO-CR * or - COCR * where R 9 = C 1-4 alkyl, branched or unbranched or phenyl; R 7, R 3 and R 4 are independently from each other hydrogen, C 1-3 alkyl, unsubstituted or substituted with one or more C 1-3 alkyl groups; k is 1-4; m is 1 to 4; p is 1-3; q is 0 or 2; or pharmaceutically acceptable salts or optical isomers thereof. A fibrinogen receptor antagonist of formula I according to claim 1, wherein X is 6 Z and C02R2 Y and Ο II where D = -0-, -S-, or -C- wherein n is 1, 2 or 3; R and R 1 are independently selected from the group consisting of phenyl, thiophene, imidazole, naphthyl, indole, indazole, 2,3-benzothiophene, unsubstituted or also substituted with hydroxy, halogen, hydroxycarbonyl-C 0-5 alkyl, C 1 alkyl unsubstituted or substituted with one or more groups which may be aryl, aryloxy, C 1-6 alkoxy, C 0 -alkylaminocarbonyl, aryl-C 8 -alkylaminocarbonyl, hydrogen, C 1 -C 6 unsubstituted or substituted or more groups which may be halogen, hydroxy, C 1-5 alkylcarbonylamino, aro-C 1-6 alkylcarbonylamino, aryloxy, C 1-6 alkyl, arylloxy, C 1-5 alkyl or aryl; C 1-5 alkoxy, C 1-5 alkoxylcarbonyl, C 1-5 alkylaminocarbonyl, C 1-6 alkylcarbonyloxy, C 3-10 Cycloalkyl, aryl, oxo, amino, C 1-6 alkyl, C 1-6 alkylamino, aminoC 1-3 alkyl, aryl Alkyl-aminocarbonyl, phenyl-C 1-4 alkylamino, aminocarbonyl-C 0-4 alkyl, or hydroxycarbonylCo-5 alkyl, provided that the carbon atom to which the R and R &quot; * groups are attached is attached to only one heteroatom. ; R 2 is hydrogen; R 7, R 3 and R 4 are hydrogen; k is 1-2; m is 1-4; p is 2-4; or the pharmaceutically acceptable salts or optical isomers thereof. The compounds of claim 1 for use in inhibiting fibrinogen binding to platelets, inhibiting platelet aggregation, treating thrombus formation and embolism, or inhibiting thrombus and embolism formation in a mammal. 5. The compounds of claim 1, selected from the group consisting of: COOH 8 / ΗΝ \ Ν COOH ΝΗ Ο 9 LV10425 Γ / HN \ N NH COOH O co2h 10th or pharmaceutically acceptable salts thereof. 6. The connection according to claim 5, which is SCH, COOH, or pharmaceutically acceptable salts thereof. 7. The compound of claim 5, which is or pharmaceutically acceptable salts thereof. 11 LV10425 8. The compound of claim 5, which is or pharmaceutically acceptable salts thereof. The compound of claim 5, which is OH COOH or a pharmaceutically acceptable salt thereof. The compound of claim 5, which is / HN V or pharmaceutically acceptable salts thereof. The compound of claim 1, which is 12th / ΗΝ \ COjH Or a pharmaceutically acceptable salt thereof. A composition for inhibiting fibrinogen binding to mammalian platelets, comprising a compound of claim 1 and a pharmaceutically acceptable excipient. 13. A composition for inhibiting platelet aggregation in a mammal comprising the compound of claim 1. A composition for inhibiting platelet aggregation in a mammal comprising a compound of claim 1 together with a thrombolytic agent and a pharmaceutically acceptable excipient. A composition for inhibiting platelet aggregation in a mammal comprising a compound of claim 1 together with an anticoagulant and a pharmaceutically acceptable excipient. A composition for the prevention and / or treatment of thrombus and embolism in a mammal comprising a compound of claim 1 and a pharmaceutically acceptable excipient. A composition for preventing and / or treating thrombus and embolism in a mammal comprising a compound of claim 1 together with a thrombolytic agent and a pharmaceutically acceptable excipient. 18. The composition of claim 14 and claim 17, wherein the thrombolytic agent is a plasminogen activator or streptokinase. 13 LV10425 A composition for preventing and / or treating thrombus and embolism formation in a mammal comprising a compound of claim 1 together with an anticoagulant and a pharmaceutically acceptable excipient. 20. The composition of claim 15 and claim 19, wherein the anticoagulant is heparin or warfarin. A composition for the treatment of thrombi and embolisms in a mammal comprising a compound of claim 1 together with an anti-platelet agent and a pharmaceutically acceptable carrier. 22. The composition of claim 21, wherein the anti-platelet agent is aspirin. Use of a composition according to claim 12 for the manufacture of a medicament for the inhibition of fibrinogen binding to platelets in mammals. Use of the compositions of claims 13, 14 and 15 for the manufacture of a medicament for the inhibition of platelet aggregation in a mammal. Use of the compositions of claims 16, 17 and 18 for the manufacture of a medicament for the inhibition of thrombus and embolism and / or for the treatment of it in a mammal. Use of a composition according to claim 21 for the manufacture of a medicament for the treatment of thrombi and embolisms in a mammal.