NZ503389A - Vitronectin receptor antagonists comprising substituted dibenzo[a,d]cycloheptene or dibenzo[b,f]oxepine and their use as antagonists of alpha-v-beta-3 receptors or alpha-v-beta-5 receptors - Google Patents

Vitronectin receptor antagonists comprising substituted dibenzo[a,d]cycloheptene or dibenzo[b,f]oxepine and their use as antagonists of alpha-v-beta-3 receptors or alpha-v-beta-5 receptors

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Publication number
NZ503389A
NZ503389A NZ503389A NZ50338998A NZ503389A NZ 503389 A NZ503389 A NZ 503389A NZ 503389 A NZ503389 A NZ 503389A NZ 50338998 A NZ50338998 A NZ 50338998A NZ 503389 A NZ503389 A NZ 503389A
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New Zealand
Prior art keywords
dibenzo
dihydro
cycloheptene
acetic acid
propyloxy
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NZ503389A
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William E Bondinell
William H Miller
Dirk Heerding
James Martin Samanen
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Smithkline Beecham Corp
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Publication of NZ503389A publication Critical patent/NZ503389A/en

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    • C07D213/60—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
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    • A61P19/00—Drugs for skeletal disorders
    • A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
    • A61P19/10—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease for osteoporosis
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
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    • C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
    • C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
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    • C07D233/28—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having one double bond between ring members or between a ring member and a non-ring member with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
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    • C07D239/08—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having one double bond between ring members or between a ring member and a non-ring member with hetero atoms directly attached in position 2
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    • C07D263/54—Benzoxazoles; Hydrogenated benzoxazoles
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    • C07D277/20—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members
    • C07D277/32—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
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    • C07D405/06—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings linked by a carbon chain containing only aliphatic carbon atoms
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    • C07D405/12—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings linked by a chain containing hetero atoms as chain links
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    • C07D487/04—Ortho-condensed systems

Abstract

A compound of formula (I), wherein: A is CH2 or O; R1 is H, halo or alkyl; R2 is H, alkyl or CH2NR"R"; X is O or CH2; Y is of the formula (II), (III), (IV), (V), (VI), (VII) or (VIII); wherein G is NR", S or O; R' is H, alkyl, -Oalkyl, -Salkyl, NR"R" or halo; each R" independently is H or alkyl; and s is 0, 1 or 2. Pharmaceutical compositions comprising the compounds of formula (I) are also described. The compounds of formula are useful for the treatment of disease in which antagonism of the anb3 receptor or the anb5 receptor is indicated.

Description

<div class="application article clearfix" id="description"> <p class="printTableText" lang="en">New Zealand Paient Spedficaiion for Paient Number 503389 <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> TITLE <br><br> Vitronectin Receptor Antagonists <br><br> FIELD OF THE INVENTION <br><br> 5 This invention relates to pharmaceutically active compounds which inhibit the vitronectin receptor and are useful for the treatment of inflammation, cancer and cardiovascular disorders, such as atherosclerosis and restenosis, and diseases wherein bone resorption is a factor, such as osteoporosis. <br><br> 0 BACKGROUND OF THE INVENTION <br><br> Integnns are a superfamily of cell adhesion receptors, which are transmembrane glycoproteins expressed on a variety of cells. These cell surface adhesion receptors include gpllb /Ilia (the fibrinogen receptor) and avB3 (the vitronectin receptor) The fibrinogen receptor gpllb /JjJa is expressed on the platelet surface, and mediates platelet aggregation 5 and the formation of a hemostatic clot at the site of a bleeding wound Philips, et al, Blood, 1988, 77, 831 The vitronectin receptor avB3 is expressed on a number of cells, including endothelial, smooth muscle, osteoclast, and tumor cells, and, thus, it has a variety of functions The avB3 receptor expressed on the membrane of osteoclast cells mediates the adhesion of osteoclasts to the bone matrix, a key step in the bone resorption process Ross, :0 et al., J Biol Chem, 1987, 262, 7703 A disease characterized by excessive bone resorption is osteoporosis The otyGj receptor expressed on human aortic smooth muscle cells mediates their migration into neointima, a process which can lead to restenosis after percutaneous coronary angioplasty. Brown, et al, Cardiovascular Res, 1994, 28, 1815 Additionally, Brooks, et al.. Cell, 1994, 79,1157 has shown that an avB3 antagonist is able 15 to promote tumor regression by inducing apoptosis of angiogenic blood vessels Thus, agents that block the vitronectin receptor would be useful in treating diseases, such as osteoporosis, restenosis and cancer <br><br> The vitronectin receptor is now known to refer to three different integnns, designatdd'a^iSj, av63 and av65 Horton, et al., Int. J Exp Pathol ,1990, 7/, 741. avBj &gt;0 binds fibronectin and vitronectin avB3 binds a large variety of ligands, including fibrin, fibrinogen, laminin, thrombospondin, vitronectin, von Willebrand's factor, osteopontm and bone sialoprotem I avBg binds vitronectin. The vitronectin receptor avBg has been shown to be involved in cell adhesion of a variety of cell types, including microvascular endothelial cells, (Davis, et al, J Cell Biol, 1993,51, 206), and its role in angiogenesis 15 has been confirmed Brooks, et al, Science, 1994,264, 569. This mtegnn is expressed on blood vessels in human wound granulation tissue, but not in normal skin <br><br> -1 - <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> The vitronectin receptor is known to bind to bone matrix proteins which contain the tn-peptide Arg-Gly-Asp (or RGD) motif. Thus, Horton, et al, Exp Cell Res 1991,195, 368, disclose that RGD-containing peptides and an anti-vitronectin receptor antibody (23C6) inhibit dentine resorption and cell spreading by osteoclasts In addition, Sato, et al, 5 J Cell Biol. 1990, 111, 1713 discloses that echistatin, a snake venom peptide which contains the RGD sequence, is a potent inhibitor of bone resorption in tissue culture, and inhibits attachment of osteoclasts to bone. <br><br> It has now been discovered that certain compounds are potent inhibitors of the &lt;xvB3 and avBj receptors. In particular, it has been discovered that such compounds are 0 more potent inhibitors of the vitronectin receptor than the fibrinogen receptor <br><br> SUMMARY OF THE INVENTION <br><br> This invention comprises compounds of the formula (I) as described hereinafter, which have pharmacological activity for the inhibition of the vitronection receptor and are 5 useful in the treatment of inflammation, cancer and cardiovascular disorders, such as atherosclerosis and restenosis, and diseases wherein bone resorption is a factor, such as osteoporosis. <br><br> This invention is also a pharmaceutical composition comprising a compound according to formula (I) and a pharmaceutically earner 0 This invention is also a method of treating diseases which are mediated by the vitronectin receptor In a particular aspect, the compounds of this invention are useful for treating atherosclerosis, restenosis, inflammation, cancer and diseases wherein bone resorption is a factor, such as osteoporosis. <br><br> 5 DETAILED DESCRIPTION <br><br> This invention comprises novel compounds which are more potent inhibitors of the vitronectin receptor than the fibrinogen receptor. The novel compounds compnse a dibenzocycloheptene core in which a nitrogen-containing substituent is present on one of the aromatic six-membered rings of the dibenzocycloheptene and an aliphatic substituent i0 containing an acidic moiety is present on the seven-membered ring of the dibenzocycloheptene The dibenzocycloheptene ring system is believed to onent the substituent sidechains on the six and seven membered rings so that they may interact favorably with the vitronectin receptor It is preferred that about twelve to fourteen intervening covalent bonds via the shortest intramolecular path will exist between the 15 acidic group on the aliphatic substituent of the seven-membered ring of the dibenzocycloheptene and the nitrogen of the nitrogen-containing substituent on one of the aromatic six-membered ring of the dibenzocycloheptene <br><br> -2- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> This invention comprises compounds of formula (I), y— (CH2)2.3—x <br><br> C0*H (i) <br><br> wherein. <br><br> 10 <br><br> A is CH2 or O, <br><br> R1 is H, halo or Cj.galkyl, R2 is H, Ci_6alkyl or CH2NR"R", X is O or CH2, <br><br> Y is <br><br> R"R"N —^ jT <br><br> O: <br><br> r-NYNR'" <br><br> ^NR" <br><br> R"R"t\L <br><br> X, <br><br> 15 <br><br> N NR''- <br><br> NL ^NR" <br><br> R"R"N <br><br> or <br><br> 20 <br><br> 25 <br><br> G is NR", S or O; <br><br> R' is H, Ci-6alkyl, OC].6alkyl, SC]_6alkyl, NR"R" or halo, <br><br> each R" independently is H or Cj.^alkyl; and s is 0,1 or 2; <br><br> or a pharmaceutically acceptable salt thereof. <br><br> Also included in this invention are pharmaceutically acceptable addition salts and complexes of the compounds of this invention In cases wherein the compounds of this invention may have one or more chiral centers, unless specified, this invention includes each unique nonracemic compound which may be synthesized and resolved by conventional techniques In cases in which compounds have unsaturated carbon-carbon <br><br> -3- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> double bonds, both the cis (Z) and trans (E) isomers are within the scope of this invention <br><br> In cases wherein compounds may exist in tautomeric forms, such as keto-enol tautomers, O OR' <br><br> such as and , and each tautomeric form is contemplated as being included within this invention whether existing in equilibrium or locked in one form by appropriate substitution with R' <br><br> The compounds of formula (I) inhibit the binding of vitronectin and other RGD-containing peptides to the vitronectin receptor. Inhibition of the vitronectin receptor on osteoclasts inhibits osteoclastic bone resorption and is useful in the treatment of diseases wherein bone resorption is associated with pathology, such as osteoporosis and osteoarthritis <br><br> In another aspect, this invention is a method for stimulating bone formation which comprises administering a compound which causes an increase in osteocalcin release. Increased bone production is a clear benefit in disease states wherein there is a deficiency of mineralized bone mass or remodeling of bone is desired, such as fracture healing and the prevention of bone fractures Diseases and metabolic disorders which result in loss of bone structure would also benefit from such treatment For instance, hyperparathyroidism, Paget's disease, hypercalcemia of malignancy, osteolytic lesions produced by bone metastasis, bone loss due to immobilization or sex hormone deficiency, Behcet's disease, osteomalacia, hyperostosis and osteopetrosis, could benefit from administering a compound of this invention. <br><br> Additionally, since the compounds of the instant invention inhibit vitronectin receptors on a number of different types of cells, said compounds would be useful in the treatment of inflammatory disorders, such as rheumatoid arthritis and psoriasis, and cardiovascular diseases, such as atherosclerosis and restenosis The compounds of Formula (I) of the present invention may be useful for the treatment or prevention of other diseases including, but not limited to, thromboembolic disorders, asthma, allergies, adult respiratory distress syndrome, graft versus host disease, organ transplant rejection, septic shock, eczema, contact dermatitis, inflammatory bowel disease, and other autoimmune diseases. The compounds of the present invention may also be useful for wound healing <br><br> The compounds of the present invention are also useful for the treatment, including prevention, of angiogenic disorders The term angiogenic disorders as used herein includes conditions involving abnormal neovascularization. Where the growth of new blood vessels is the cause of, or contributes to, the pathology associated with a disease, inhibition of angiogenisis will reduce the deleterious effects of the disease An example of such a disease target is diabetic retinopathy. Where the growth of new blood vessels is required to support growth of a deleterious tissue, inhibition of angiogenisis will reduce the blood <br><br> -4- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> supply to the tissue and thereby contribute to reduction in tissue mass based on blood supply requirements Examples include growth of tumors where neovascularization is a continual requirement in order that the tumor grow and the establishment of solid tumor metastases Thus, the compounds of the present invention inhibit tumor tissue 5 angiogenesis, thereby preventing tumor metastasis and tumor growth <br><br> Thus, according to the methods of the present invention, the inhibition of angiogenesis using the compounds of the present invention can ameliorate the symptoms of the disease, and, in some cases, can cure the disease. <br><br> 10 diseases chacterized by neovascularization. Such eye diseases include corneal neovascular disorders, such as corneal transplantation, herpetic keratitis, luetic keratitis, pterygium and neovascular pannus associated with contact lens use Additional eye diseases also include age-related macular degeneration, presumed ocular histoplasmosis, retinopathy of prematurity and neovascular glaucoma 15 This invention further provides a method of inhibiting tumor growth which comprises administering stepwise or in physical combination a compound of formula (I) and an antineoplastic agent, such as topotecan and cisplatin <br><br> , wherein R' is H, Cj^alkyl, OC^alkyl, SC].4alkyl, NR"R" or CI and each R" independently is H or Cj^alkyl Alternately, Y is <br><br> Another therapeutic target for the compounds of the instant invention are eye <br><br> With respect to formula (I) Suitably Y is <br><br> 20 <br><br> N. ,NR" <br><br> 25 <br><br> , wherein each R" is H or Cj^alkyl. <br><br> Alternately, Y is <br><br> N _ . NR" <br><br> ().v^nr- <br><br> , wherein each R" independently is H or Cj^alkyl, <br><br> 30 and s is 1 <br><br> -5- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> Alternately, Y is <br><br> R"R"N <br><br> G <br><br> , wherein G is S and each R" independently is H or <br><br> C]_4alkyl <br><br> Alternately, Y is <br><br> , wherein R" is H or Cj^alkyl. <br><br> Representative of the novel compounds of this invention are the following (±)-10,11 -Dihydro-3-[2-(6-aminopyridin-2-yl)-1 -ethoxy]-5H-dibenzo[a,d]cycloheptene-10-acetic acid; <br><br> (±)-10,11 -Dihydro-3-[4-(pyndin-2-ylamino)-1 -buty 1]-5H-dibenzo[a,d]cycloheptene-10-acetic acid, <br><br> (±)-10,11 -Dihydro-3-[3-(4-ethoxypyridm-2-ylamino)-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetic acid; <br><br> (S)-10,11 -Dihydro-3-[3-(pyndin-2-y lamino)-1 -propyloxy ]-5H-dibenzo[a,d]cycloheptene- 10-acetic acid; <br><br> (R)-10,l l-Dihydro-3-[3-(pyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetic acid; <br><br> (±)-10,11 -Dihydro-3- [3-(3,4,5,6-tetrahydropyrimidin-2-ylamino)-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetic acid, <br><br> (±)-10,11 -Dihydro-3-[2-[2-(ethylamino)thiazol-4-yl]-l -ethoxy]-5H-dibenzo[a,d]cycloheptene-10-acetic acid; <br><br> (±)-10,11 -Dihydro-3-[3-(isoquinoline-1 -ylamino)-1 -propy loxy]-5H-dibenzo[a,d]cycloheptene-10-acetic acid, <br><br> (±)-10,11 -Dihydro-7-fluoro-3-[3-(pyridin-2-ylamino)-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene- 10-acetic acid, <br><br> (S)-10,11 -Dihydro-3-[3-(4-methylpyridin-2-ylamino)-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetic acid, <br><br> (S)-10,11 -Dihydro-3-[3-(4-ethoxypyndin-2-ylamino)-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene- 10-acetic acid, <br><br> (+)-10,11 -Dihydro-6-methyl-3-[3-(pyridin-2-ylamino)- l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetic acid. <br><br> -6- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> (±)-10,11 -Dihydro-2-(dimethylamino)methyl-7-fluoro-3-[3-(pyndin-2-ylamino)-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetic acid, <br><br> (S)-10,ll-Dihydro-3-[3-[4-(2-propyloxy)pyridin-2-ylamino]-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetic acid, <br><br> 5 (S)-10,11 -Dihy dro-3- [2- [6-(methylamino)pyndin-2-y 1]-1 -ethoxy ]-5H- <br><br> dibenzo[a,d]cycloheptene-10-acetic acid, <br><br> (S)-10,11 -Dihydro-3-[3-[4-(dimethylamino)pyridin-2-y lamino]-1 -propy loxy )-5H-dibenzo[a,d]cycloheptene-10-acetic acid, <br><br> (±)-10,11 -Dihy dro-3- [3- [4-(ethy lthio)pyndin-2-y lammo]-1 -propy loxy ]-5H-10 dibenzo[a,d]cycloheptene-10-acetic acid; <br><br> (S)-10,11 -Dihydro-3-[3-(4-chloropyridm-2-yl amino)-1 -propy loxy ]-5H-dibenzo[a,d]cycloheptene- 10-acetic acid, <br><br> (+)-10,11 -Dihydro-2-methyl-3-[3-(pyridin-2-ylamino)-1 -propyloxy ]-5H-dibenzo[a,d]cycloheptene-10-acetic acid, <br><br> 15 (S)-10,11 -Dihydro-3-[3-(4-aminopyndin-2-ylamino)-1 -propyloxy]-5H- <br><br> dibenzo[a,d]cycloheptene-10-acetic acid; <br><br> (±)-10,11 -Dihydro-3-[3-(4-methylpyndin-2-ylamino)-1 -propyloxy]-dibenzo[b,f]oxepine-10-acetic acid, <br><br> (±)-10,11 -Dihydro-3-[2-[6-(methylamino)pyridin-2-yl]-1 -ethoxy ]-20 dibenzo[b,f]oxepine-10-acetic acid; and <br><br> (S)-10,11 -Dihydro-3-[3-(2-aminopyridin-4-yl)-1 -propyloxy]-5//-dibenzo[a,&lt;/]cycloheptene-10-acetic acid; <br><br> or a pharmaceutically acceptable salt thereof. <br><br> In cases wherein the compounds of this invention may have one or more chiral 25 centers, unless specified, this invention includes each unique nonracemic compound which may be synthesized and resolved by conventional techniques According to the present invention, the (S) configuration of the formula (I) compounds is preferred <br><br> In cases in which compounds have unsaturated carbon-carbon double bonds, both the cis (Z) and trans (E) isomers are within the scope of this invention The meaning of any 30 substituent at any one occurrence is independent of its meaning, or any other substituent's meaning, at any other occurrence. <br><br> Also included in this invention are prodrugs of the compounds of this invention. Prodrugs are considered to be any covalently bonded carriers which release the active parent drug according to formula (I) in vivo Thus, in another aspect of this invention are 35 novel prodrugs, which are also intermediates in the preparation of formula (I) compounds, of formula (II). <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> Y— (CH2)2.3 X <br><br> C02Ci.6alkyl <br><br> (II) <br><br> wherein: <br><br> A is CH2 or O; <br><br> R1 is H, halo or Cj.galkyl; R2 is H, C].6alkyl or CH2NR"R", X is O or CH2, <br><br> Y is <br><br> 10 <br><br> R"R"N —^ || «— <br><br> r-NYNR"" <br><br> R"R"NL -N. <br><br> H <br><br> R"R"N <br><br> or <br><br> 15 GisNR", SorO, <br><br> R' is H, C]-6alkyl, OCi-galkyl, SCi-6alkyl, NR"R" or halo, <br><br> each R" independently is H or Cj.galkyl; and s is 0,1 or 2, <br><br> or a pharmaceutically acceptable salt thereof 20 In yet another aspect of this invention are novel intermediates of formula (III): <br><br> -8- <br><br> Pnnted from Mimosa <br><br> WO 99/15508 PCT/US98/19466 <br><br> wherein. <br><br> A is CH2 or O, <br><br> R' is H, halo orCj.galkyl; <br><br> 5 R2 is H, C].6alkyl or CH2NR"R", <br><br> X is O or CH2, <br><br> R' is H, C].6alkyl, OCj.galkyl, SCi_6alkyl, NR"R" or halo, and each R" independently is H or Chalky I, <br><br> or a pharmaceutically acceptable salt thereof 10 Abbreviations and symbols commonly used in the peptide and chemical arts are used herein to describe the compounds of this invention. In general, the amino acid abbreviations follow the IUPAC-IUB Joint Commission on Biochemical Nomenclature as described in Eur. J Biochem., 158, 9 (1984). <br><br> Cj_4alkyl as applied herein means an optionally substituted alkyl group of 1 to 4 15 carbon atoms, and includes methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl and t-butyl Cj.galkyl additionally includes pentyl, n-pentyl, isopentyl, neopentyl and hexyl and the simple aliphatic isomers thereof. C^alkyl and Cg^alkyl additionally indicates that no alkyl group need be present (e g , that a covalent bond is present). <br><br> Any C^alkyl or C]_g alkyl may be optionally substituted with the group Rx, 20 which may be on any carbon atom that results in a stable structure and is available by conventional synthetic techniques. Suitable groups for Rx are Cj^alkyl, OR", SR , CMalkylsulfonyl, C^alkylsulfoxyl, -CN, N(R")2, CH2N(R")2, -N02, -CF3, -CO2R", -CON(R")2, -COR", -NR"C(0)R", F, CI, Br, I, or CF3S(0)r-, wherein r is 0, 1 or 2 Halogen or halo means F, CI, Br, and I. <br><br> 25 Ar, or aryl, as applied herein, means phenyl or naphthyl, or phenyl or naphthyl substituted by one to three substituents, such as those defined above for alkyl, especially C^alkyl, Cj.4alkoxy, Cj^alkthio, CF3, NH2, OH, F, CI, Br or I <br><br> Certain radical groups are abbreviated herein t-Bu refers to the tertiary butyl radical, Boc refers to the t-butyloxycarbonyl radical, Fmoc refers to the 30 fluorenylmethoxycarbonyl radical, Ph refers to the phenyl radical, Cbz refers to the benzyloxycarbonyl radical, Bn refers to the benzyl radical, Me refers to methyl, Et refers to <br><br> -9- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/U S98/19466 <br><br> ethyl, Ac refers to acetyl, Alk refers to C^alkyl, Nph refers to 1- or 2-naphthyl and cHex refers to cyclohexyl Tet refers to 5-tetrazolyl <br><br> Certain reagents are abbreviated herein DCC refers to dicyclohexylcarbodnmide, DMAP refers to dimethylaminopyridine, DIEA refers to dnsopropylethyl amine, EDC refers to l-(3-dimethylaminopropyl)-3-ethylcarbodnmide, hydrochloride HOBt refers to 1-hydroxybenzotnazole, THF refers to tetrahydrofuran, DIEA refers to diisopropylethylamine, DEAD refers to diethyl azodicarboxylate, PPh3 refers to triphenylphosphine, DIAD refers to dusopropyl azodicarboxylate, DME refers to dimethoxyethane, DMF refers to dimethylformamide, NBS refers to N-bromosuccinimide, Pd/C refers to a palladium on carbon catalyst, PPA refers to polyphosphonc acid, DPPA refers to diphenylphosphoryl azide, BOP refers to benzotriazol-l-yloxy-tris(dimethyl-amino)phosphomum hexafluorophosphate, HF refers to hydrofluoric acid, TEA refers to triethylamine, TFA refers to trifluoroacetic acid, PCC refers to pyndimum chlorochromate. <br><br> The compounds of formula (I) are generally prepared by reacting a compound of formula (IV) with a compound of formula (V). <br><br> and thereafter removing any protecting groups, and optionally forming a pharmaceutically acceptable salt <br><br> Suitably, certain compounds of formula (I) are prepared by reacting a compound of formula (IV) with a compound of formula (VI): <br><br> HO <br><br> C02C16alkyl Y-(CH2)2.3-Ll <br><br> (IV) <br><br> (V) <br><br> wherein R^, R2, Y and A are as defined in formula (I), with any reactive functional groups protected, and l' is OH or halo, <br><br> HO <br><br> O- <br><br> l + <br><br> ,Nt.NRM (CH2)2.3 —OH <br><br> (IV) <br><br> (VI) <br><br> - 10- <br><br> Prmted from Mimosa <br><br> WO 99/15508 PCT/US98/19466 <br><br> wherein R*, R2, R', R" and A are as defined in formula (I), with any reactive functional groups protected, <br><br> and thereafter removing any protecting groups, and optionally forming a pharmaceutically acceptable salt 5 Suitably, the reaction between a compound of formual (IV) with a compound of formula (VI) is carried out in the presence of diethyl azodicarboxylate and tnphenylphosphme in an aprotic solvent <br><br> Additionally, certain compounds of formula (I) are prepared by reacting a compound of formula (IV) with a compound of formula (VII) <br><br> 10 <br><br> O- <br><br> R" | <br><br> /N N+ ^(CH2)2.3—OH R" ^ <br><br> (IV) (VII) <br><br> wherein R^, R2, R" and A are as defined in formula (I), with any reactive 15 functional groups protected; <br><br> and thereafter removing any protecting groups, and optionally forming a pharmaceutically acceptable salt <br><br> Suitably, the reaction between a compound of formual (IV) with a compound of formula (VII) is carried out in the presence of diethyl azodicarboxylate and 20 tnphenylphosphme in an aprotic solvent. <br><br> Compounds of the formula (I) are prepared by the methods described in Bondmell et al, PCT Publication No WO 97/01540 (International Application No. PCT/US96/11108), published January 16,1997, the entire disclosure of which is incorporated herein by reference 25 Additionally, compounds of formula (I) are prepared by methods analogous to those descnbed in the schemes that are detailed hereinafter <br><br> -11 - <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> Scheme 1 <br><br> 1 2 3 <br><br> a) 10% Pd/C, HO Ac; b) SOCl2, toluene; c) A1C1V CH2C12 <br><br> 5 <br><br> Scheme I details the preparation of an intermediate useful in the preparation of formula (I) compounds <br><br> - 12- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> Scheme II <br><br> d, e <br><br> CO,Et a) LiN(TMS)2, ethyl bromoacetate, b) Jones reagent, 0s04; c) H2, 10% Pd/C, HOAC, d) C202C12, DMF, e) A1C13, CH2C12, RT; f) H2, 10% Pd/C, HOAC <br><br> 5 Scheme II also details the preparation of an intermediate useful in the preparation of formula (I) compounds <br><br> - 13- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCTAJS98/19466 <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> (a) EtOAc/LiHMDS, THF; (b) H2, 10% Pd/C, conc HCl, AcOH, (c) EtSH, AICI3, CH2C12, (d) 2-t(3-hydroxy-l-propyl)amino]-4-nitropyridine-N-oxide, DEAD, (Ph^P, (e) NaOEt, EtOH; (f) cyclohexene, 10% Pd/C, EtOH; (g) 1 0 N NaOH, EtOH, (h) HCl <br><br> 5 Scheme III details the preparation of a formula (I) compound Reaction of III-l <br><br> (which is a Scheme 1-3 compound) in an aldol-type reaction with the enolate of ethyl acetate, which can be generated from ethyl acetate on exposure to an appropriate amide base, for instance lithium dusopropylamide (LDA) or lithium bis(trimethylsilyl)amide (LiHMDS), gives III-2 Frequently, THF is the solvent of choice for an aldol reaction, 10 although THF in the presence of various additives, for instance HMPA or TMEDA, is often used. Reduction of III-2 to give III-3 (which is a Scheme II-6 compound) can be accomplished by hydrogenolysis over an appropriate catalyst, for example palladium metal on activated carbon (Pd/C), in an appropriate solvent, such as acetic acid, in the presence of a mineral acid such as HCl Alternatively, this reduction can be accomplished by treatment 15 of III-2 with tnethylsilane in the presence of boron trifluonde etherate by the general method of Orfanopoulos and Smonou (Synth Commun. 1988, 833) Removal of the methyl ether of III-3 to give III-4 can be accomplished with BBr3 in an inert solvent, for example CH2CI2, or by reaction with ethanethiol and AICI3 in an inert solvent, preferably CH2CI2. Other useful methods for removal of a methyl ether are described in Greene, "Protective 20 Groups in Organic Synthesis" (published by John Wiley and Sons) Compound 4 of <br><br> Scheme 3 (III-4) is reacted with 2-[(3-hydroxy-l-propyl)amino]-4-mtropyndine-N-oxide in a Mitsunobu-type coupling reaction (Organic Reactions 1992,42, 335-656, Synthesis 1981, 1 -28) to afford III-5 The reaction is mediated by the complex formed between diethyl azodicarboxylate and tnphenylphosphme, and is conducted in an aprotic solvent, for 25 instance THF, CH2CI2, or DMF Compound III-5 is reacted with an alkali metal salt of an appropriate alcohol to afford III-6 Suitable alkali metals include lithium, sodium, potassium, and cesium, and the alcohol used for the displacement reaction is generally used as the solvent. Methods for forming the alkali metal salts of alcohols are well-known to those of skill m the art. The pyndme-N-oxide moiety of III-6 is reduced to the 30 corresponding pyndine III-7 under transfer hydrogenation conditions using a palladium catalyst, preferably palladium metal on activated carbon, in an inert solvent, for instance methanol, ethanol, or 2-propanol Cyclohexene, 1,4-cyclohexadiene, formic acid, and salts of formic acid, such as potassium formate or ammonium formate, are commonly used as the hydrogen transfer reagent in this type of reaction. The ethyl ester of III-7 is hydrolyzed 35 using aqueous base, for example, LiOH in aqueous THF or NaOH in aqueous methanol or ethanol, and the intermediate carboxylate salt is acidified with a suitable acid, for instance TFA or HCl, to afford the carboxylic acid III-8 Alternatively, the intermediate <br><br> -15- <br><br> Pnnted from Mimosa <br><br> WO 99/15508 <br><br> PCT /US98/19466 <br><br> carboxylate salt can be isolated, if desired, or a carboxylate salt of the free carboxyhc acid can be prepared by methods well-known to those of skill in the art <br><br> Scheme IV <br><br> C02Et <br><br> C02Et <br><br> (a) NaH, 2-[N-(3-methanesulfonyloxy-1 -propyl)-N-(rert-butoxycarbonyl)amino]pyndine-N-oxide, DMSO, (b) TFA, CH2CI2, (c) see Scheme III <br><br> 10 <br><br> Scheme IV describes an alternative method for the preparation of formula (I) compounds. Compound IV-1 is reacted with a base, preferably an alkali metal hydride such as sodium hydride or potassium hydride, in a polar, aprotic solvent, generally THF, DMF, DMSO, or mixtures thereof, to afford the corresponding alkali metal phenoxide. <br><br> 15 Alternatively, an alkali metal amide, for instance LDA, or the lithium, sodium, or potassium salt of hexamethyldisilazane, can be used for deprotonation. The intermediate phenoxide is generally not isolated, but is reacted in situ with an appropriate electrophile, for instance 2-[N-(3-methanesulfonyloxy-l-propyl)-N-(rerf-butoxycarbonyl)-amino]pyndine-N-oxide, to afford the coupled product IV-2. The /ert-butoxycarbonyl <br><br> 20 protecting group in IV-2 is removed under acidic conditions, such as 4 M HCl in 1,4- <br><br> - 16- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> dioxane or TFA in CH2CI2, to afford IV-3 Conditions for removal of the tert-butoxycarbonyl protecting group are well-known to those of skill in the art, and several useful methods are described in standard reference volumes such as Greene "Protective Groups in Organic Synthesis" IV-3 is subsequently converted to IV-4 following the 5 procedure outlined in Scheme III <br><br> Scheme V <br><br> MeO <br><br> MeO <br><br> 6 HO ^—COaEt <br><br> 9.h <br><br> 10 <br><br> MeO <br><br> COjEt <br><br> (a) PhOH, Cu, K2C03, (b) sulfur, morpholine; (c) KOH, H20, i-PrOH, (d) SOCl2, benzene, (e) AICI3, CH2C12, (f) EtOAc, LiN(TMS)2, TMEDA, THF, (g) Et3SiH, BF3 OEt2, CH2C12; (h) H2, Pd/C, EtOH; (i) BBr3&gt; CH2C12 <br><br> 15 Commercially available 2-fluoro-4-methoxyacetophenone (V-l) reacts with an alcohol, for example phenol, in the presence of copper metal and a suitable base, for instance K2CO3, to afford the diaryl ether V-2 On treatment with sulfur and an appropriate primary or secondary amine, preferably morpholine, according to the general <br><br> -17- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> method of Harris (J Med Chem 1982, 25, 855), V-2 is converted to V-3 in a classical Willgerodt-Kindler reaction The thioamide thus obtained is hydrolyzed to the corresponding carboxyhc acid V-4 by reaction with an alkali metal hydroxide, suitably KOH, in an aqueous alcoholic solvent, such as aqueous MeOH, EtOH, or 1-PrOH 5 Carboxyhc acid V-4 is converted to the corresponding acid chlonde by reaction with either SOCI2 or oxalyl chloride according to conditions well-known to those of skill in the art Treatment of this acid chloride with an appropriate Friedel-Crafts catalyst, such as AICI3 or SnCl4, in an inert solvent, such as CH2CI2 or CS2, provides the cyclic ketone V-5 Alternatively, acid V-4 can be converted directly to ketone V-5 under acidic conditions, for 10 example with polyphosphonc acid Reaction of V-5 in an aldol - type reaction with the enolate of ethyl acetate, which can be generated from ethyl acetate on exposure to an appropriate amide base, for instance lithium dusopropylamide (LDA) or lithium bis(trimethylsilyl)amide (LiHMDS), gives V-6 Frequently, THF is the solvent of choice for an aldol reaction, although THF in the presence of various additives, for instance 15 HMPA or TMEDA, is often used Reduction of V-6 to give V-7 can be accomplished by treatment of V-6 with triethylsilane in the presence of boron trifluonde etherate by the general method of Orphanopoulos and Smonu (Synth Commun. 1988, 833). Any olefinic by-products that result from elimination of the alcohol are reduced by hydrogenation over an appropriate catalyst, for example palladium metal on activated carbon (Pd/C), in an 20 appropriate solvent, such as MeOH or EtOH. Alternatively, the reduction of V-6 to give V-7 can be accomplished by hydrogenolysis in the presence of a mineral acid such as HCl Typically, this reaction is catalyzed by Pd/C, and is optimally conducted in acetic acid. Removal of the methyl ether of V-7 to give V-8 can be accomplished with BBr3 in an inert solvent, for example CH2CI2, or by reaction with ethanethiol and AICI3 in an inert solvent, 25 preferably CH2CI2 Other useful methods for removal of a methyl ether are described in Greene, "Protective Groups in Organic Synthesis" (published by John Wiley and Sons). V-8 is subsequently converted to formula (I) compounds following the procedure outlined in Scheme III. <br><br> Acid addition salts of the compounds are prepared in a standard manner in a 30 suitable solvent from the parent compound and an excess of an acid, such as hydrochloric, hydrobromic, hydrofluoric, sulfuric, phosphoric, acetic, tnfluoroacetic, maleic, succinic or methanesulfonic. Certain of the compounds form inner salts or zwitterions which may be acceptable. Cationic salts are prepared by treating the parent compound with an excess of an alkaline reagent, such as a hydroxide, carbonate or alkoxide, containing the appropriate 35 cation, or with an appropriate organic amine Cations such as Li+, Na+, K+, Ca++, Mg++ and NH4+ are specific examples of cations present in pharmaceutically acceptable salts <br><br> - 18- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT /US98/19466 <br><br> This invention also provides a pharmaceutical composition which comprises a compound according to formula (I) and a pharmaceutically acceptable carrier. <br><br> Accordingly, the compounds of formula (I) may be used in the manufacture of a medicament Pharmaceutical compositions of the compounds of formula (I) prepared as hereinbefore described may be formulated as solutions or lyophilized powders for parenteral administration Powders may be reconstituted by addition of a suitable diluent or other pharmaceutically acceptable carrier prior to use. The liquid formulation may be a buffered, isotonic, aqueous solution. Examples of suitable diluents are normal isotonic saline solution, standard 5% dextrose in water or buffered sodium or ammonium acetate solution Such formulation is especially suitable for parenteral administration, but may also be used for oral administration or contained m a metered dose inhaler or nebulizer for insufflation It may be desirable to add excipients such as polyvinylpyrrolidone, gelatin, hydroxy cellulose, acacia, polyethylene glycol, mannitol, sodium chloride or sodium citrate <br><br> Alternately, these compounds may be encapsulated, tableted or prepared in a emulsion or syrup for oral administration Pharmaceutically acceptable solid or liquid carriers may be added to enhance or stabilize the composition, or to facilitate preparation of the composition Solid carriers include starch, lactose, calcium sulfate dihydrate, terra alba, magnesium stearate or stearic acid, talc, pectin, acacia, agar or gelatin Liquid carriers include syrup, peanut oil, olive oil, saline and water The carrier may also include a sustained release material such as glyceryl monostearate or glyceryl distearate, alone or with a wax The amount of solid earner varies but, preferably, will be between about 20 mg to about 1 g per dosage unit The pharmaceutical preparations are made following the conventional techniques of pharmacy involving milling, mixing, granulating, and compressing, when necessary, for tablet forms; or milling, mixing and filling for hard gelatin capsule forms When a liquid carrier is used, the preparation will be in the form of a syrup, elixir, emulsion or an aqueous or non-aqueous suspension Such a liquid formulation may be administered directly p.o or filled into a soft gelatin capsule. <br><br> For rectal administration, the compounds of this invention may also be combined with excipients such as cocoa butter, glycenn, gelatin or polyethylene glycols and molded into a suppository <br><br> The compounds described herein are antagonists of the vitronectin receptor, and are useful for treating diseases wherein the underlying pathology is attnbutable to ligand or cell which interacts with the vitronectin receptor. For instance, these compounds are useful for the treatment of diseases wherein loss of the bone matrix creates pathology. Thus, the instant compounds are useful for the treatment of ostoeporosis, hyperparathyroidism, <br><br> Paget's disease, hypercalcemia of malignancy, osteolytic lesions produced by bone <br><br> -19- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> metastasis, bone loss due to immobilization or sex hormone deficiency The compounds of this invention are also believed to have utility as antitumor, anti-angiogenic, antiinflammatory and anti-metastatic agents, and be useful in the treatment of atherosclerosis and restenosis. <br><br> 5 The compound is administered either orally or parenterally to the patient, in a manner such that the concentration of drug is sufficient to inhibit bone resorption, or other such indication The pharmaceutical composition containing the compound is administered at an oral dose of between about 0.1 to about 50 mg/kg in a manner consistent with the condition of the patient. Preferably the oral dose would be about 0 5 to about 20 mg/kg. 0 For acute therapy, parenteral administration is preferred An intravenous infusion of the peptide in 5% dextrose in water or normal saline, or a similar formulation with suitable excipients, is most effective, although an intramuscular bolus injection is also useful Typically, the parenteral dose will be about 0 01 to about 100 mg/kg; preferably between 0 1 and 20 mg/kg The compounds are administered one to four times daily at a level to 5 achieve a total daily dose of about 0 4 to about 400 mg/kg/day. The precise level and method by which the compounds are administered is readily determined by one routinely skilled in the art by comparing the blood level of the agent to the concentration required to have a therapeutic effect. <br><br> This invention further provides a method for treating osteoporosis or inhibiting 0 bone loss which comprises administering stepwise or in physical combination a compound of formula (I) and other inhibitors of bone resorption, such as bisphosphonates (i.e., alendronate), hormone replacement therapy, anti-estrogens, or calcitonin In addition, this invention provides a method of treatment using a compound of this invention and an anabolic agent, such as the bone morphogenic protein, lproflavone, useful in the prevention :5 of bone loss and/or to increase bone mass <br><br> Additionally, this invention provides a method of inhibiting tumor growth which comprises administering stepwise or in physical combination a compound of formula (I) and an antineoplastic agent Compounds of the camptothecm analog class, such as topotecan, lnnotecan and 9-aminocamptothecin, and platinum coordination complexes, 10 such as cisplatm, ormaplatin and tetraplatin, are well known groups of antineoplastic agents Compounds of the camptothecm analog class are described in U S Patent Nos 5,004,758,4,604,463,4,473,692,4,545,880 4,342,776,4,513,138,4,399,276, EP Patent Application Publication Nos 0 418 099 and 0 088 642, Warn, et al, J. Med Chem , 1986, 29, 2358, Warn, et al., J. Med Chem , 1980,23,554, Warn, et al., J Med Chem , 1987,30, i5 1774, and Nitta, et al., Proc 14th International Congr Chemotherapy., 1985, Anticancer Section 1, 28, the entire disclosure of each which is hereby incorporated by reference. The platinum coordination complex, cisplatm, is available under the name Platmol® from <br><br> -20- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/U S98/19466 <br><br> Bristol Myers-Squibb Corporation Useful formulations for cisplatm are described in U S Patent Nos 5,562,925 and 4,310,515, the entire disclosure of each which is hereby incorporated by reference. <br><br> In the method of inhibiting tumor growth which comprises administering stepwise or in physical combination a compound of formula (I) and an antineoplastic agent, the platinum coordination compound, for example cisplatm, can be administered using slow intravenous infusion The preferred carrier is a dextrose/saline solution containing mannitol. The dose schedule of the platinum coordination compound may be on the basis of from about 1 to about 500 mg per square meter (mg/m^) of body surface area per course of treatment Infusions of the platinum coordiation compound may be given one to two times weekly, and the weekly treatments may be repeated several times Using a compound of the camptothecin analog class in a parenteral administration, the course of therapy generally employed is from about 0 1 to about 300 0 mg/m^ of body surface area per day for about five consecutive days Most preferably, the course of therapy employed for topotecan is from about 1.0 to about 2.0 mg/m^ of body surface area per day for about five consecutive days. Preferably, the course of therapy is repeated at least once at about a seven day to about a twenty-eight day interval <br><br> The pharmaceutical composition may be formulated with both the compound of formula (I) and the antineoplastic agent in the same container, but formualtion in different containers is preferred. When both agents are provided m solution form, they can be contained in an infusion/injection system for simultaneous administration or in a tandem arrangement <br><br> For convenient administration of the compound of formula (I) and the antineoplastic agent at the same or different times, a kit is prepared, comprising, in a single container, such as a box, carton or other container, individual bottles, bags, vials or other containers each having an effective amount of the compound of formula (I) for parenteral administration, as described above, and an effective amount of the antineoplastic agent for parenteral administration, as described above Such kit can comprise, for example, both pharmaceutical agents in separate containers or the same container, optionally as lyophilized plugs, and containers of solutions for reconstitution. A variation of this is to include the solution for reconstitution and the lyophilized plug in two chambers of a single container, which can be caused to admix prior to use With such an arrangement, the antineoplastic agent and the compound of this invention may be packaged separately, as in two containers, or lyophilized together as a powder and provided in a single container <br><br> When both agents are provided in solution form, they can be contained in an infusion/injection system for simultaneous administration or m a tandem arrangement. For example, the compound of formula (I) may be in an i v injectable form, or infusion bag <br><br> -21 - <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> linked in series, via tubing, to the antineoplastic agent in a second infusion bag Using such a system, a patient can receive an initial bolus-type injection or infusion of the compound of formula (I) followed by an infusion of the antineoplastic agent <br><br> The compounds may be tested in one of several biological assays to determine the 5 concentration of compound which is required to have a given pharmacological effect <br><br> Inhibition of vitronectin binding <br><br> Solid-Phase [3HJ-SK&amp;F-107260 Binding to CCvfy Human placenta or human platelet avp3 (0.1-0 3 mg/mL) in buffer T (containing 2 mM CaCl2 and 1% octylglucoside) was 10 diluted with buffer T containing 1 mM CaCl2, 1 mM MnCl2, 1 mM MgCl2 (buffer A) and 0 05% NaN3, and then immediately added to 96-well ELISA plates (Corning, New York, NY) at 0 1 mL per well. 0.1 - 0 2 pg of CCvP3 was added per well. The plates were incubated overnight at 4°C. At the time of the experiment, the wells were washed once with buffer A and were incubated with 0 1 mL of 3.5% bovine serum albumin in the same 15 buffer for 1 hr at room temperature. Following incubation the wells were aspirated completely and washed twice with 0 2 mL buffer A <br><br> Compounds were dissolved in 100% DMSO to give a 2 mM stock solution, which was diluted with binding buffer (15 mM Tns-HCl (pH 7.4), 100 mM NaCl, 1 mM CaCl2, 1 mM MnCl2,1 mM MgCl2) to a final compound concentration of 100 |iM. This solution is 20 then diluted to the required final compound concentration. Vanous concentrations of unlabeled antagonists (0.001 - 100 pM) were added to the wells in triplicates, followed by the addition of 5 0 nM of [^HJ-SK&amp;F-107260 (65 - 86 Ci/mmol). <br><br> The plates were incubated for 1 hr at room temperature. Following incubation the wells were aspirated completely and washed once with 0 2 mL of ice cold buffer A in a 25 well-to-well fashion The receptors were solubilized with 0.1 mL of 1% SDS and the bound [-^HJ-SK&amp;F-107260 was determined by liquid scintillation counting with the addition of 3 mL Ready Safe in a Beckman LS Liquid Scintillation Counter, with 40% efficiency Nonspecific binding of [^H]-SK&amp;F-107260 was determined in the presence of 2 |iM SK&amp;F-107260 and was consistently less than 1% of total radioligand input The 30 IC50 (concentration of the antagonist to inhibit 50% binding of pHJ-SK&amp;F-107260) was determined by a nonlinear, least squares curve-fitting routine, which was modified from the LUNDON-2 program The K] (dissociation constant of the antagonist) was calculated according to the equation- K] = IC50/O + L/K&lt;j), where L and Kd were the concentration and the dissociation constant of [^HJ-SK&amp;F-107260, respectively 35 Compounds of the present invention inhibit vitronectin binding to SK&amp;F 107260 in the concentration range of about 2 5 to about 0 001 micomolar <br><br> -22- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> Compounds of this invention are also tested for in vitro and in vivo bone resorption in assays standard m the art for evaluating inhibition of bone formation, such as the pit formation assay disclosed in EP 528 587, which may also be performed using human osteoclasts in place of rat osteoclasts, and the ovarectomized rat model, described by 5 Wronski et al, Cells and Materials 1991, Sup 1,69-74. <br><br> Vascular smooth muscle cell migration assay <br><br> Rat or human aortic smooth muscle cells were used. The cell migration was monitored in a Transwell cell culture chamber by using a polycarbonate membrane with 0 pores of 8 um (Costar) The lower surface of the filter was coated with vitronectin. Cells were suspended in DMEM supplemented with 0 2% bovine serum albumin at a concentration of 2 5 - 5 0 x 106 cells/mL, and were pretreated with test compound at various concentrations for 20 min at 20°C The solvent alone was used as control 0.2 mL of the cell suspension was placed in the upper compartment of the chamber. The lower 5 compartment contained 0 6 mL of DMEM supplemented with 0 2% bovine serum albumin Incubation was earned out at 37°C in an atmosphere of 95% air/5% CO2 for 24 hr After incubation, the non-migrated cells on the upper surface of the filter were removed by gentle scraping. The filter was then fixed in methanol and stained with 10% Giemsa stain. <br><br> Migration was measured either by a) counting the number of cells that had migrated to the 0 lower surface of the filter or by b) extracting the stained cells with 10% acetic acid followed by determining the absorbance at 600 nM. <br><br> Thyroparathyroidectomized rat model <br><br> Each experimental group consists of 5-6 adult male Sprague-Dawley rats (250-400g body 5 weight) The rats are thyroparathyroidectomized (by the vendor, Taconic Farms) 7 days prior to use All rats receive a replacement dose of thyroxine every 3 days. On receipt of the rats, circulating ionized calcium levels are measured in whole blood immediately after it has been withdrawn by tail venipuncture into hepannized tubes Rats are included if the ionized Ca level (measured with a Ciba-Coming model 634 calcium pH analyzer) is &lt;1.2 mM/L. Each rat is fitted 0 with an indwelling venous and arterial catheter for the delivery of test material and for blood sampling respectively The rats are then put on a diet of calcium-free chow and deionized water. Baseline Ca levels are measured and each rat is administered either control vehicle or human parathyroid hormone 1-34 peptide (hPTHl-34, dose 1.25 ug/kg/h in saline/0.1% bovine serum albumin, Bachem, Ca) or a mixture of hPTHl-34 and test material, by continuous intravenous '5 infusion via the venous catheter using an external syringe pump. The calcemic response of each rat is measured at two-hourly intervals during the infusion period of 6-8 hours <br><br> -23- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> Human osteoclast resorption and adhesion assays <br><br> Pit resorption and adhesion assays have been developed and standardized using normal human osteoclasts derived from osteoclastoma tissue. Assay 1 was developed for the measurement of osteoclast pit volumes by laser confocal microscopy Assay 2 was 5 developed as a higher throughput screen in which collagen fragments (released during resorption) are measured by competitve ELISA. <br><br> Assay 1 (using laser confocal microscopy) <br><br> • Aliquots of human osteoclastoma-derived cell suspensions are removed from liquid 0 nitrogen strorage, warmed rapidly at 37°C and washed xl in RPMI-1640 medium by centrifugation (lOOOrpm, 5 mins at 4°C) <br><br> • The medium is aspirated and replaced with murine anti-HLA-DR antibody then diluted 1 3 in RPMI-1640 medium. The suspension is incubated for 30 mins on ice and mixed frequently <br><br> 5 • The cells are washed x2 with cold RPMI-1640 followed by centrifugation (1000 rpm, 5 mins at 4°C) and the cells are then transferred to a sterile 15 ml centrifuge tube The number of mononuclear cells are enumerated in an improved Neubauer counting chamber. <br><br> • Sufficient magnetic beads (5 / mononuclear cell), coated with goat anti-mouse IgG 0 (Dynal, Great Neck, NY) are removed from their stock bottle and placed into 5 ml of fresh medium (this washes away the toxic azide preservative) The medium is removed by immobilizing the beads on a magnet and is replaced with fresh medium <br><br> • The beads are mixed with the cells and the suspension is incubated for 30 mins on ice. The suspension is mixed frequently. <br><br> :5 • The bead-coated cells are immobilized on a magnet and the remaining cells (osteoclast-rich fraction) are decanted into a sterile 50 ml centrifuge tube. <br><br> • Fresh medium is added to the bead-coated cells to dislodge any trapped osteoclasts This wash process is repeated xlO. The bead-coated cells are discarded. <br><br> • The viable osteoclasts are enumerated in a counting chamber, using fluorescein t0 diacetate to label live cells A large-bore disposable plastic pasteur pipet is used to add the sample to the chamber <br><br> • The osteoclasts are pelleted by centrifugation and the density adjusted to the appropriate number in EMEM medium (the number of osteoclasts is variable from tumor to tumor), supplemented with 10% fetal calf serum and 1 7g/hter of sodium <br><br> 15 bicarbonate <br><br> • 3ml aliquots of the cell suspension (per compound treatment) are decanted into 15ml centrifuge tubes The cells are pelleted by centrifugation <br><br> -24- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> • To each tube, 3ml of the appropriate compound treatment are added (diluted to 50 uM in the EMEM medium) Also included are appropriate vehicle controls, a positive control (anti-vitronectm receptor murine monoclonal antibody [87MEM1] diluted to 100 ug/ml) and an isotype control (IgG2j diluted to 100 ug/ml) The samples are <br><br> 5 incubated at 37°C for 30 mins. <br><br> • 0 5ml aliquots of the cells are seeded onto sterile dentine slices in a 48-well plate and incubated at 37°C for 2 hours Each treatment is screened in quadruplicate <br><br> • The slices are washed in six changes of warm PBS (10 ml / well in a 6-well plate) and then placed into fresh medium containing the compound treatment or control <br><br> 0 samples The samples are incubated at 37°C for 48 hours <br><br> Tartrate resistant acid phosphatase (TRAP) procedure (selective stain for cells of the osteoclast lineage) <br><br> • The bone slices containing the attached osteoclasts are washed in phosphate <br><br> 5 buffered saline and fixed in 2% gluteraldehyde (m 0.2M sodium cacodylate) for 5 mins <br><br> • They are then washed in water and are incubated for 4 minutes in TRAP buffer at 37°C (0 5 mg/ml naphthol AS-BI phosphate dissolved m N,N-dimethylformamide and mixed with 0.25 M citrate buffer (pH 4.5), containing 10 mM sodium tartrate. <br><br> • Following a wash in cold water the slices are immersed in cold acetate buffer (0 1 <br><br> 0 M, pH 6 2) containing 1 mg/ml fast red garnet and incubated at 4°C for 4 minutes <br><br> • Excess buffer is aspirated, and the slices are air dried following a wash in water <br><br> • The TRAP positive osteoclasts (brick red/ purple precipitate) are enumerated by bright-field microscopy and are then removed from the surface of the dentine by sonication. <br><br> .5 • Pit volumes are determined using the Nikon/Lasertec 1LM21W confocal microscope <br><br> Assay 2 (using an ELISA readout) <br><br> The human osteoclasts are enriched and prepared for compound screening as <br><br> &gt;0 described in the initial 9 steps of Assay 1 For clanty, these steps are repeated hereinbelow. <br><br> • Aliquots of human osteoclastoma-denved cell suspensions are removed from liquid nitrogen strorage, warmed rapidly at 37°C and washed xl in RPMI-1640 medium by centrifugation (lOOOrpm, 5 mins at 4°C) <br><br> • The medium is aspirated and replaced with murine anti-HLA-DR antibody then <br><br> 15 diluted 1 3 in RPMI-1640 medium. The suspension is incubated for 30 mins on ice and mixed frequently. <br><br> -25- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> • The cells are washed x2 with cold RPMI-1640 followed by centrifugation (1000 rpm, 5 mins at 4°C) and the cells are then transferred to a sterile 15 ml centrifuge tube The number of mononuclear cells are enumerated in an improved Neubauer counting chamber <br><br> 5 • Sufficient magnetic beads (5 / mononuclear cell), coated with goat anti-mouse IgG (Dynal, Great Neck, NY) are removed from their stock bottle and placed into 5 ml of fresh medium (this washes away the toxic azide preservative). The medium is removed by immobilizing the beads on a magnet and is replaced with fresh medium. <br><br> • The beads are mixed with the cells and the suspension is incubated for 30 mins on <br><br> 0 ice. The suspension is mixed frequently <br><br> • The bead-coated cells are immobilized on a magnet and the remaining cells (osteoclast-rich fraction) are decanted into a sterile 50 ml centrifuge tube. <br><br> • Fresh medium is added to the bead-coated cells to dislodge any trapped osteoclasts. This wash process is repeated xlO. The bead-coated cells are discarded. <br><br> 5 • The viable osteoclasts are enumerated in a counting chamber, using fluorescein diacetate to label live cells A large-bore disposable plastic pasteur pipet is used to add the sample to the chamber <br><br> • The osteoclasts are pelleted by centrifugation and the density adjusted to the appropriate number in EMEM medium (the number of osteoclasts is variable from <br><br> 0 tumor to tumor), supplemented with 10% fetal calf serum and 1.7g/liter of sodium bicarbonate <br><br> In contrast to the method desnbed above m Assay 1, the compounds are screened at <br><br> 4 doses to obtain an IC5„, as outlined below <br><br> • The osteoclast preparations are preincubated for 30 minutes at 37°C with test <br><br> .5 compound (4 doses) or controls <br><br> • They are then seeded onto bovine cortical bone slices in wells of a 48-well tissue culture plate and are incubated for a further 2 hours at 37°C <br><br> • The bone slices are washed in six changes of warm phosphate buffered saline (PBS), to remove non-adherent cells, and are then returned to wells of a 48 well plate <br><br> 0 containing fresh compound or controls <br><br> • The tissue culture plate is then incubated for 48 hours at 37°C <br><br> • The supernatants from each well are aspirated into individual tubes and are screened in a competitive ELISA that detects the c-telopeptide of type I collagen which is released during the resorption process This is a commercially available ELISA <br><br> 15 (Osteometer, Denmark) that contains a rabbit antibody that specifically reacts with an 8-amino acid sequence (Glu-Lys-Ala-His- Asp-Gly-Gly-Arg) that is present in the carboxy-terminal telopeptide of the al-chain of type I collagen The results are <br><br> -26- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> expressed as % inhibition of resorption compared to a vehicle control <br><br> Human osteoclast adhesion assay <br><br> The human osteoclasts are enriched and prepared for compound screening as 5 described above in the mital 9 steps of Assay 1. For clarity, these steps are repeated hereinbelow. <br><br> • Aliquots of human osteoclastoma-denved cell suspensions are removed from liquid nitrogen strorage, warmed rapidly at 37°C and washed xl in RPMI-1640 medium by centrifugation (lOOOrpm, 5 mms at 4°C) <br><br> 10 • The medium is aspirated and replaced with murine anti-HLA-DR antibody then diluted 1:3 in RPMI-1640 medium The suspension is incubated for 30 mins on ice and mixed frequently <br><br> • The cells are washed x2 with cold RPMI-1640 followed by centrifugation (1000 rpm, 5 mms at 4°C) and the cells are then transferred to a sterile 15 ml centrifuge tube. <br><br> 15 The number of mononuclear cells are enumerated m an improved Neubauer counting chamber <br><br> • Sufficient magnetic beads (5 / mononuclear cell), coated with goat anti-mouse IgG (Dynal, Great Neck, NY) are removed from their stock bottle and placed into 5 ml of fresh medium (this washes away the toxic azide preservative) The medium is removed <br><br> 20 by immobilizing the beads on a magnet and is replaced with fresh medium <br><br> • The beads are mixed with the cells and the suspension is incubated for 30 mms on ice. The suspension is mixed frequently. <br><br> • The bead-coated cells are immobilized on a magnet and the remaining cells (osteoclast-rich fraction) are decanted into a sterile 50 ml centrifuge tube <br><br> 25 • Fresh medium is added to the bead-coated cells to dislodge any trapped osteoclasts This wash process is repeated xlO The bead-coated cells are discarded <br><br> • The viable osteoclasts are enumerated in a counting chamber, using fluorescein diacetate to label live cells. A large-bore disposable plastic pasteur pipet is used to add the sample to the chamber <br><br> 30 • The osteoclasts are pelleted by centrifugation and the density adjusted to the appropriate number in EMEM medium (the number of osteoclasts is variable from tumor to tumor), supplemented with 10% fetal calf serum and 1 7g/liter of sodium bicarbonate <br><br> • Osteoclastoma-denved osteoclasts are preincubated with compound (4 doses) or <br><br> 35 controls at 37"C for 30 minutes <br><br> • The cells are then seeded onto osteopontm-coated slides (human or rat osteopontin, 2.5ug/ml) and incubated for 2 hours at 37°C. <br><br> -27- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> • Non adherent cells are removed by washing the slides vigorously in phosphate buffered saline and the cells remaining on the slides are fixed in acetone <br><br> • The osteoclasts are stained for tartrate-resistant acid phosphatase (TRAP), a selective marker for cells of this phenotype (see steps 15 -17), and are enumerated by <br><br> 5 light microscopy The results are expressed as % inhibition of adhesion compared to a vehicle control. <br><br> Cell Adhesion Assay Cells and Cell Culture <br><br> Human embryonic kidney cells (HEK293 cells) were obtained from ATCC 0 (Catalog No. CRL 1573). Cells were grown in Earl's minimal essential medium (EMEM) medium containing Earl's salts, 10% fetal bovine serum, 1% glutamine and 1% Penicillm-Steptomycin. <br><br> Constructs and Transfections <br><br> A 3 2 kb EcoRI-Kpnl fragment of the av subumt and a 2.4 kb Xbal- Xhol fragment 5 of the P3 subumt were inserted into the EcoRI - EcoRV cloning sites of the pCDN vector (Aiyar et al, 1994) which contains a CMV promoter and a G418 selectable marker by blunt end ligation For stable expression, 80 x 10 6 HEK 293 cells were electrotransformed with av+P3 constructs (20 |Ag DNA of each subunit) using a Gene Pulser (Hensley et al, 1994) and plated in 100 mm plates (5x10^ cells/plate) After 48 hr, the growth medium .0 was supplemented with 450 )ig/mL Geneticm (G418 Sulfate, GIBCO-BRL, Bethesda, MD). The cells were maintained in selection medium until the colonies were large enough to be assayed <br><br> Immunocytochemical analysis of transfected cells <br><br> To determine whether the HEK 293 transfectants expressed the vitronectin :5 receptor, the cells were immobilized on glass microscope slides by centrifugation, fixed m acetone for 2 mm at room temperature and air dried Specific reactivity with 23C6, a monoclonal antibody specific for the avP3 complex was demonstrated using a standard indirect immunofluorescence method. <br><br> 10 Cell Adhesion Studies <br><br> Corning 96-well ELISA plates were precoated overnight at 4°C with 0 1 mL of human vitronectin (0 2 (J.g/mL in RPMI medium). At the time of the experiment, the plates were washed once with RPMI medium and blocked with 3 5% BSA in RPMI medium for 1 hr at room temperature. Transfected 293 cells were resuspended in RPMI medium, <br><br> -28- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> supplemented with 20 mM Hepes, pH 7 4 and 0.1% BSA at a density of 0 5 x 10^ cells/mL 0 1 mL of cell suspension was added to each well and incubated for 1 hr at 37°C, in the presence or absence of various avp3 antagonists Following incubation, 0 025 mL of a 10% formaldehyde solution, pH 7 4, was added and the cells were fixed at room 5 temperature for 10 mm. The plates were washed 3 times with 0.2 mL of RPMI medium and the adherent cells were stained with 0.1 mL of 0.5% toluidine blue for 20 min at room temperature. Excess stain was removed by extensive washing with deionized water The toluidine blue incorporated into cells was eluted by the addition of 0.1 mL of 50% ethanol containing 50 mM HCl Cell adhesion was quantitated at an optical density of 600 nm on a 10 microtiter plate reader (Titertek Multiskan MC, Sterling, VA) <br><br> Solid-Phase avp5 Binding Assay: <br><br> The vitronectin receptor avps was purified from human placenta. Receptor preparation was diluted with 50 mM Tns-HCl, pH 7.5,100 mM NaCl, 1 mM CaCl2,1 mM 15 MnCl2, 1 mM MgCl2 (buffer A) and was immediately added to 96-well ELISA plates at 0 1 ml per well 0 1 -0.2 (jg of avP3 was added per well The plates were incubated overnight at 4°C At the time of the experiment, the wells were washed once with buffer A and were incubated with 0 1 ml of 3.5% bovine serum albumin in the same buffer for 1 hr at room temperature Following incubation the wells were aspirated completely and washed twice 20 with 0.2 ml buffer A <br><br> In a [3H]-SK&amp;F-107260 competition assay, various concentrations of unlabeled antagonists (0 001-100 pM) were added to the wells, followed by the addition of 5.0 nM of [3H]-SK&amp;F-107260. The plates were incubated for 1 hr at room temperature Following incubation the wells were aspirated completely and washed once with 0 2 ml of ice cold 25 buffer A in a well-to-well fashion The receptors were solubilized with 0 1 ml of 1 % SDS and the bound [3H]-SK&amp;F-107260 was determined by liquid scintillation counting with the addition of 3 ml Ready Safe in a Beckman LS 6800 Liquid Scintillation Counter, with 40% efficiency. Nonspecific binding of [3H]-SK&amp;F-107260 was determined in the presence of 2 |iM SK&amp;F-107260 and was consistently less than 1% of total radioligand input The IC50 30 (concentration of the antagonist to inhibit 50% binding of [3H]-SK&amp;F-107260) was determined by a nonlinear, least squares curve-fitting routine, which was modified from the LUNDON-2 program The K, (dissociation constant of the antagonist) was calculated according to Cheng and Prusoff equation K, = IC50/ (1 + L/Kj), where L and K&lt;j were the concentration and the dissociation constant of [3H]-SK&amp;F-107260, respectively. <br><br> 35 <br><br> -29- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT7US98/19466 <br><br> Inhibition of RGD-mediated GPIIb-IIIa binding <br><br> Purification of GPIIb-IIIa <br><br> Ten units of outdated, washed human platelets (obtained from Red Cross) were 5 lyzed by gentle stirring m 3% octylglucoside, 20 mM Tns-HCl, pH 7 4, 140 mM NaCl, 2 mM CaCl2 at 4°C for 2 h The lysate was centrifuged at 100,000g for 1 h. The supernatant obtained was applied to a 5 mL lentil lectin sepharose 4B column (E Y Labs) preequilibrated with 20 mM Tns-HCl, pH 7.4, 100 mM NaCl, 2 mM CaCl2, 1% octylglucoside (buffer A) After 2 h incubation, the column was washed with 50 mL cold 10 buffer A. The lectin-retained GPIIb-IIIa was eluted with buffer A containing 10% dextrose. All procedures were performed at 4°C The GPIIb-IIIa obtained was &gt;95% pure as shown by SDS polyacrylamide gel electrophoresis <br><br> Incorporation of GPIIb-IIIa in Liposomes 15 A mixture of phosphatidylsenne (70%) and phosphatidylcholine (30%) (Avanti <br><br> Polar Lipids) were dried to the walls of a glass tube under a stream of nitrogen Purified GPIIb-IIIa was diluted to a final concentration of 0 5 mg/mL and mixed with the phospholipids in a protein phospholipid ratio of 1:3 (ww). The mixture was resuspended and sonicated in a bath sonicator for 5 mm. The mixture was then dialyzed overnight using 20 12,000-14,000 molecular weight cutoff dialysis tubing against a 1000-fold excess of 50 mM Tns-HCl, pH 7.4,100 mM NaCl, 2 mM CaC12 (with 2 changes). The GPIIb-IIIa-contaimng liposomes wee centnfuged at 12,000g for 15 min and resuspended in the dialysis buffer at a final protein concentration of approximately 1 mg/mL The liposomes were stored at -70C until needed <br><br> 25 <br><br> Competitive Binding to GPIIb-IIIa <br><br> The binding to the fibrinogen receptor (GPIIb-IIIa) was assayed by an indirect competitive binding method using [^HJ-SK&amp;F-107260 as an RGD-type hgand The binding assay was performed in a 96-well filtration plate assembly (Millipore Corporation, 30 Bedford, MA) using 0 22 um hydrophilic durapore membranes. The wells were precoated with 0 2 mL of 10 (ig/mL polylysine (Sigma Chemical Co., St. Louis, MO.) at room temperature for 1 h to block nonspecific binding. Various concentrations of unlabeled benzazepines were added to the wells in quadruplicate [^Hl-SK&amp;F-107260 was applied to each well at a final concentration of 4 5 nM, followed by the addition of 1 pg of the purified 35 platelet GPIIb-IIIa-containing liposomes The mixtures were incubated for 1 h at room temperature The GPIIb-IIIa-bound [3H]-SK&amp;F-107260 was seperated from the unbound by filtration using a Millipore filtration manifold, followed by washing with ice-cold buffer <br><br> -30- <br><br> Prmted from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> (2 times, each 0 2 mL) Bound radioactivity remaining on the filters was counted in 1 5 mL Ready Solve (Beckman Instruments, Fullerton, CA) in a Beckman Liquid Scintillation Counter (Model LS6800), with 40% efficiency Nonspecific binding was determined in the presence of 2 |iM unlabeled SK&amp;F-107260 and was consistently less than 0.14% of the 5 total radioactivity added to the samples All data points are the mean of quadruplicate determinations <br><br> Competition binding data were analyzed by a nonlinear least-squares curve fitting procedure. This method provides the IC50 of the antagonists (concentration of the antagonist which inhibits specific binding of [3H]-SK&amp;F-107260 by 50% at equilibrium) 10 The IC50 is related to the equilibrium dissociation constant (Ki) of the antagonist based on the Cheng and Prusoff equation. Ki = IC50/(l+L/Kd), where L is the concentration of [3H]-SK&amp;F-107260 used in the competitive binding assay (4.5 nM), and Kd is the dissociation constant of [3H]-SK&amp;F-107260 which is 4.5 nM as determined by Scatchard analysis <br><br> Preferred compounds of this invention have an affinity for the vitronectin receptor 15 relative to the fibrinogen receptor of greater than 10 1. Most preferred compounds have a ratio of activity of greater than 100:1 <br><br> The efficacy of the compounds of formula (I) alone or in combination with an antineoplastic agent may be determined using several transplantable mouse tumor models 20 See U S Patent Nos 5,004,758 and 5,633,016 for details of these models <br><br> The examples which follow are intended in no way to limit the scope of this invention, but are provided to illustrate how to make and use the compounds of this invention. Many other embodiments will be readily apparent to those skilled in the art <br><br> 25 <br><br> -31 - <br><br> Printed from Mimosa <br><br> WO 99/15508 PCT/US98/19466 <br><br> EXAMPLES <br><br> General <br><br> 5 Proton nuclear magnetic resonance ('H NMR) spectra were recorded at either 250 <br><br> or 400 MHz Chemical shifts are reported in parts per million (8) downfield from the internal standard tetramethylsilane (TMS). Abbreviations for NMR data are as follows. s=singlet, d=doublet, t=tnplet, q=quartet, m=multiplet, dd=doublet of doublets, dt=doublet of triplets, app=apparent, br=broad. J indicates the NMR coupling constant measured in 10 Hertz CDCI3 is deuteriochloroform, DMSO-d6 is hexadeuteriodimethylsulfoxide, and CD3OD is tetradeutenomethanol Infrared (IR) spectra were recorded in transmission mode, and band positions are reported in inverse wavenumbers (cm"'). Mass spectra were obtained using electrospray (ES) or FAB ionization techniques Elemental analyses were performed either m-house or by Quantitative Technologies Inc , Whitehouse, NJ. Melting 15 points were taken on a Thomas-Hoover melting point apparatus and are uncorrected All temperatures are reported in degrees Celsius Analtech Silica Gel GF and E. Merck Silica Gel 60 F-254 thin layer plates were used for thin layer chromatography Both flash and gravity chromatography were earned out on E. Merck Kieselgel 60 (230-400 mesh) silica gel. Analytical and preparative HPLC were carried out on Rainin or Beckman 20 chromatographs. ODS refers to an octadecylsilyl denvatized silica gel chromatographic support 5 p Apex-ODS indicates an octadecylsilyl denvatized silica gel chromatographic support having a nominal particle size of 5 p, made by Jones Chromatography, Littleton, Colorado YMC ODS-AQ® is an ODS chromatographic support and is a registered trademark of YMC Co. Ltd, Kyoto, Japan. PRP-1® is a polymeric (styrene-25 divinylbenzene) chromatographic support, and is a registered trademark of Hamilton Co., Reno, Nevada Cehte® is a filter aid composed of acid-washed diatomaceous silica, and is a registered trademark of Manville Corp., Denver, Colorado. <br><br> Ethyl (±)-10,l l-dihydro-3-methoxy-5H-dibenzo[a,d]cycloheptene-10-acetate, ethyl (±)-10,l l-dihydro-3-hydroxy-5H-dibenzo[a,d]cycloheptene-10-acetate, and ethyl (±)-30 10,ll-dihydro-3-(tnfluoromethanesulfonyloxy)-5H-dibenzo[a,d]cycloheptene-10-acetate were prepared according to WO 9701540-A1. 2-[2-(4-Methoxybenzylammo)pyndin-6-yljethanol was prepared according to WO 95/32710 6-Methoxy-l-indanone was prepared by the method of House and Hudson (J. Org Chem. 1970,35,647). <br><br> -32- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCMJS98/19466 <br><br> Preparation 1 <br><br> Preparation of 2-r(3-hvdroxv-l-pronvl')aminolpvndine-N-oxide <br><br> 5 a) 2-[(3-Hydroxy-l-propyl)amino]pyndme-N-oxide <br><br> A mixture of 2-chloropyndine-N-oxide hydrochloride (16 6 g, 0 1 mole), 3-amino-1-propanol (15.3 mL, 0.2 mole), NaHCC&gt;3 (42 g, 0.5 mole), and rerr-amyl alcohol (100 mL) was heated to reflux After 21 hr, the reaction was cooled, diluted with CH2CI2 (300 mL), and suction filtered to remove insoluble matenals. The filtrate was concentrated and 10 reconcentrated from toluene to leave a yellow oil. Silica gel chromatography (20% MeOH/CHCl3) gave the title compound (15.62 g, 93%) as a yellow solid: TLC (20% MeOH/CHCl3) Rf 0.48, &gt;H NMR (250, CDCI3) 8 8 07 (dd, J = 6 6, 1.2 Hz, 1 H), 7 34 (br t, 1 H), 7 10 - 7 30 (m, 1 H), 6 64 (dd, J = 8 5, 1.4 Hz, 1 H), 6 40 - 6.60 (m, 1 H),449(brs, 1 H), 3 65 - 3.90 (m, 2 H), 3 35 - 3 60 (m, 2 H), 1 75 - 2 00 (m, 2 H); MS (ES) m/e 169 (M+ 15 H)+ <br><br> Preparation 2 <br><br> Preparation of 2-f(3-hvdroxv-1 -pronvllaminol-4-nitropvridine-N-oxide <br><br> 20 <br><br> a) 2-Chloro-4-nitropyridine-N-oxide <br><br> A solution of conc H2SO4 (30 mL) and fuming HNO3 (54 mL) was added dropwise at 0°C to a solution of 2-chloropyridine-N-oxide hydrochloride (15 2 g, 91 56 mmole) in conc. H2SO4 (30 mL) The reaction mixture was heated at 90°C for 1 hr, then 25 was cooled to RT and poured onto ice (500 g) The reaction mixture was kept at RT overnight, then was cooled in an ice bath, and 50% NaOH was added slowly to give a precipitate This was collected and dried to give the title compound (5.88 g, 37 %) as a pale yellow solid ]H NMR (400 MHz, CDCI3) 8 8.42 - 8 37 (m, 2 H), 8.06 -8.04 (m, 1 H) <br><br> 30 a) 2-[(3-Hydroxy-l-propyl)amino]-4-nitropyndine-N-oxide <br><br> According to the procedure of Preparation 1, except substituting 2-chloro-4-nitropyndme-N-oxide for the 2-chloropyridine-N-oxide hydrochloride, the title compound was obtained as yellow powder following silica gel chromatography (1 9 MeOH/CH2Cl2) Recrystallization from Me0H/CH2Cl2/Et20 gave the title compound- MS (ES) 214 1 35 (M+H)+ <br><br> -33- <br><br> Printed from Mimosa <br><br> WO 99/15508 PCT/US98/19466 <br><br> Preparation 3 <br><br> Preparation of 2-[C3-hvdroxv-l-DropvnaminoT-4-methvlpvridine-N-oxide <br><br> 5 a) 2-Chloro-4-methylpyridine <br><br> Sodium nitnte (13.88 g, 200 mmole) was added slowly at 0°C to a solution of 2-amino-4-picohne (15 0 g, 139 mmole) in conc. HCl (200 mL) The reaction mixture was allowed to warm to RT and was stirred for 16 hr, then was poured onto ice (500 g) The pH was adjusted to 8 0 with conc. NH4OH, and the mixture was extracted with ether (3 x 300 0 mL). The combined ether layers were washed sequentially with H2O (2 x 200 mL) and brine (200 mL). Drying (MgSC&gt;4) and concentration gave the title compound (10 3 g, 58%) as a faintly yellow oil MS (ES) m/e 127.8 (M + H)+ <br><br> b) 2-Chloro-4-methylpyridine-N-oxide hydrochloride <br><br> 5 A mixture of 2-chioro-4-methylpyridine (10 0 g, 78.3 mmole) and 34% peracetic acid (76 05 g, 91 0 mmole) in glacial AcOH (10 mL) was heated at 70°C for 3 hr The reaction mixture was cooled, conc HCl (35 mL) was added, and the mixture was concentrated on the rotavap Recrystallization from n-butanol followed by trituration with ether gave the title compound (7 16 g, 51%) as a white solid MS (ES) m/e 143 9 (M + ,0 H)+ <br><br> c) 2-[(3-Hydroxy-l-propyl)amino]-4-methylpyridine-N-oxide <br><br> A mixture of 2-chloro-4-methylpyridine-N-oxide hydrochloride (7 16 g, 39 mmole), 3-ammopropanol (6 01 g, 80 mmole), and NaHC03 (16 8 g, 200 mmole) in tert-:5 amyl alcohol (50 mL) was heated at reflux for 19 hr. The reaction mixture was diluted with CH2CI2 (200 mL) and filtered, and the filtrate was concentrated on the rotavap Recrystallization from CH2Cl2/Et20 gave the title compound (5 41 g, 75%) as a yellow solid" TLC (15% MeOH/CH2Cl2) Rf 0.44, 'H NMR (400, CDCI3) 8 7 92 (d, J = 6 7,1 H), 7 28 (br t, 1 H), 6 43 (s, 1 H), 6.33 (dd, J = 6.6,2.1 Hz, 1 H), 3.73 (t, J=5 7 Hz, 2 H), 3 47 -0 (q, H=6.3 Hz, 2 H), 2 29 (s, 3 H), 1.82- 1 88 (m, 2 H), MS (ES) m/e 183 (M+ H)+ <br><br> -34- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> Preparation 4 <br><br> Preparation of 6-(methvlamino)-2-pvridvlethanol <br><br> 5 a) 2-(fert-Butoxycarbonylamino)-6-picoline <br><br> A solution of 2-amino-6-picohne (21 63 g, 200 mmole) and di-ferf-butyl dicarbonate (52 38 g, 240 mmole) in CH2CI2 (200 mL) was concentrated on the rotavap at 50°C, and the resulting residue was allowed to rotate on the rotavap at 50°C under vacuum After 21.5 hr, the reaction was diluted with hexanes (400 mL) and filtered through silica gel 0 (hexanes followed by 20% EtOAc/hexanes) Concentration left the title compound (41.84 g, quantitative) as a light yellow oil which gradually solidified on standing 'H NMR (250 MHz, CDCI3) 8 7 71 (d, J = 8 3 Hz, 1 H), 7 40 - 7.65 (m, 2 H), 6 80 (d, J = 7 5 Hz, 1 H), 2 43 (s, 3 H), 1 50 (s, 9 H), MS (ES) m/e 153 (M + H - C4H8)+. <br><br> 5 b) 2-[(ferf-Butoxycarbonyl)methylamino]-6-picoline <br><br> NaH (60% in mineral oil, 3.60 g, 90 mmole) was added in portions over several min to a solution of 2-(ter/-butoxycarbonylamino)-6-picohne (15.62 g, 75 mmole) and lodomethane (9 3 mL, 150 mmole) in anhydrous DMSO (75 mL) at 15°C (cool water bath). The internal temperature rose to 35°C When gas evolution had subsided, the cool water 0 bath was removed and the reaction was allowed to stir at RT After 0 5 hr, the dark yellow mixture was poured onto ice/H20 (300 mL) and extracted with Et20 (3 x 300 mL). The combined organic layers were washed sequentially with H2O (2 x 75 mL) and brine (75 mL). Drying (MgS04) and concentration left a yellow oil which was chromatographed on silica gel (7% EtOAc/hexanes) The title compound (13.01 g, 78%) was obtained as a :5 faintly yellow oil 1H NMR (250 MHz, CDCI3) 87 51 (app t, 1 H), 7 37 (d, J = 8.2 Hz, 1 H), 6 86 (d, J = 7 2 Hz, 1 H), 3.38 (s, 3 H), 2 49 (s, 3 H), 1.50 (s, 9 H); MS (ES) m/e 223 (M + H)+. <br><br> c) Ethyl-6-[(ferf-butoxycarbonyl)methyIammo]-2-pyridylacetate 10 LDA was prepared at 0°C under argon from dnsopropylamme (19 5 mL, 139.14 <br><br> mmole) and 2.5 M n-BuLt in hexanes (46 4 mL, 115.95 mmole) in dry THF (350 mL). <br><br> This solution was cooled to -78°C and a solution of 2-[(fert-butoxycarbonyl)methylamino]-6-picoline (10 31 g, 46.38 mmole) in dry THF (46 mL) was added dropwise over 10 min Additional dry THF (2 mL) was used in transfer. The orange solution was stirred at -78°C (5 for 15 min, then diethyl carbonate (6 2 mL, 51.02 mmole) was added rapidly The red solution was stirred at -78°C for 15 min, then was quenched with half-saturated NH4CI (175 mL) The mixture was warmed to +5°C and extracted with EtOAc (175 mL) then with <br><br> -35- <br><br> Prmted from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> CH2CI2 (2 x 100 mL) The combined organics were washed with brine (100 mL). dried (MgSC&gt;4), and concentrated The cloudy yellow oil was chromatographed on silica gel (15% EtOAc/hexanes) to afford the title compound (10.72 g, 79%) as a light yellow oil ]H NMR (250 MHz, CDCI3) 8 7 51 - 7 63 (m, 2 H), 6 91 - 7 03 (m, 1 H), 4 19 (q. J = 7 1 5 Hz, 2 H), 3.77 (s, 2 H), 3 38 (s, 3 H), 1.27 (t, J = 7 1 Hz, 3 H), 1 51 (s, 9 H), MS (ES) m/e 295 (M + H)+. <br><br> d) EthyI-6-(methylamino)-2-pyridylacetate <br><br> A solution of ethyl-6-[(/err-butoxycarbonyl)methylamino]-2-pyridylacetate (10.72 10 g, 36 42 mmole) in anhydrous dioxane (91 mL) was cooled to the point of partial crystallization of the solvent, and 4 M HCl/dioxane (91 mL, 364.2 mmole) was added. The solution was warmed to RT and stirred for 17 hr, then was concentrated The resulting light yellow solid was slurried with CH2Cl2/toluene and reconcentrated to leave the title compound (8 48 g, quantitative) as a light yellow powder *H NMR (250 MHz, CD3OD) 8 15 7 84 (dd, J = 9.0, 7 2 Hz, 1 H), 6 96 (d, J = 9 0 Hz, 1 H), 6 78 (d, J = 7.2 Hz, 1 H). 4 22 (q, J = 7 1 Hz, 2 H), 3.93 (s, 2 H), 3.05 (s, 3 H), 1.27 (t, J = 7 1 Hz, 3 H), MS (ES) m/e 195 (M + H)+ <br><br> e) 6-(Methylamino)-2-pyridylethanol <br><br> 20 A solution of 1 0 M L1AIH4 in THF (95 mL, 95 mmole) was added dropwise to a mechanically stirred suspension of ethyl-2-(methylamino)-6-pyndylacetate (7.34 g, 31 82 mmole) in dry THF (64 mL) at 0°C under argon. The addition was done slowly until gas evolution subsided, then the remaining solution was added rapidly. Addition required 5-7 mm The reaction was warmed to RT and stirred for 45 min, then was heated to reflux 25 After 10 min, the reaction was cooled to 0°C and worked up by sequential dropwise addition of H2O (3 6 mL), 15% NaOH (3 6 mL), and H2O (10 8 mL) The mixture was stirred for 15 min at 0°C and 15 min at RT, then was filtered through a Buchner funnel The filter pad was washed with plenty of THF, and the filtrate was concentrated. The residue was reconcentrated from toluene, then was chromatographed on silica gel (5% 30 MeOH in 1 1 EtOAc/CHC^) to afford the title compound (3 23 g, 67%) as a yellow oil which solidified to a waxy solid. ^H NMR (250 MHz, CDCI3) 8 7.36 (dd, J = 8.3,7.3 Hz, 1 H), 6.42 (d, J = 7 3 Hz, 1 H), 6.26 (d, J = 8.3 Hz, 1 H), 4.93 - 5.28 (m, 1 H), 4.38 - 4 60 (m, 1 H), 3 96 (t, J = 5 4 Hz, 2 H), 2.90 (d, J = 5.2 Hz, 3 H), 2 84 (t, J = 5.4 Hz, 2 H), MS (ES) m/e 153 (M + H)+ <br><br> -36- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> Preparation 5 <br><br> Preparation of 2-(ethvlaminoV4-thiazoleethanol <br><br> 5 a) Ethyl 2-acetylammo-4-thiazoleacetate <br><br> Ethyl 2-amino-4-thiazoleacetate (3 72 g, 20 mmole) was taken up in acetic acid (4 mL) and acetic anhydride (4 mL), and the resulting suspension was heated at reflux for 3 hr. Concentration and flash chromatography on silica gel (5% MeOH/CH2H2) gave the title compound (4 1 g, 91%) as a white solid: MS (ES) m/e 229 (M + H)+. <br><br> 0 <br><br> b) 2-(Ethylamino)-4-thiazoleethanol <br><br> To a stirred solution of 1.0 M LiAUfy in THF (179 mL, 179 mmole) was added dropwise a solution of ethyl 2-acetylamino-4-thiazoleacetate (4 4 g .17.9 mmole) in THF (50 mL) After complete addition, the reaction mixture was heated at reflux for 3 hr, then 5 was worked up by sequential addition of H20 (0.7 mL), 10% NaOH (0 7 mL), and H2O (2 1 mL) The resulting mixture was filtered through cehte® and the filtrate was concentrated Purification by flash chromatography on silica gel (5% MeOH/CH2H2) gave the title compound (1 6 g, 53%) as an amber oil MS (ES) m/e 173 (M + H)+ <br><br> 0 Preparation 6 <br><br> Preparation of 6-amino-2-pvridvlethanol a) 6-Amino-2-pyridylethanol .5 A solution of 2-[2-(4-methoxybenzylamino)pyridin-6-yl]ethanol (0 95 g, 3 7 <br><br> mmole), prepared according to the procedure of WO 95/32710, m 6 N HCl was heated at 60°C After 16 hr, the reaction was concentrated in vacuum and the residue was made basic with dry KOH The resulting mixture was extracted with MeOH, and the MeOH extracts were dried (MgSO/j) and concentrated Flash chromatography on silica gel (5% 0 MeOH/CH2H2) gave the title compound (0.2 g, 40%) as a pale yellow oil: MS (ES) m/e 139 (M + H)+. <br><br> -37- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> Preparation 7 <br><br> Preparation of 3-(4-nitrobenzvloxvcarbonvlaminoVl-propanol <br><br> 5 a) 3-(4-Nitrobenzyloxycarbonyl)amino-l-propanol <br><br> To a suspension stirred under argon at room temperature of 4-nitrobenzyl chloroformate (5 g, 23 mmol) and tnethylamine (6 4 mL, 46 mmol) in THF (25 mL) was added 3-ammo-1 -propanol (1.9 mL, 26 mmol) The resulting mixture was stirred for 72 hr, then was concentrated The residue was purified by chromatography on silica gel (0 5 - 2% 0 MeOH/CH2Cl2) to give the title compound (2 g, 34%) as a pale yellow oil MS (ES) 255 3 (M+H)+. <br><br> Preparation 8 <br><br> 5 Preparation of 1 -Nl-hydroxy- l-propvllaminolisoqumohne-N-oxide a) 1-Chloroisoqumoline N-oxide <br><br> 1-Aminoisoquinoline N-oxide hydrochloride (Deady, L W Synthetic Communications 1977, 509-514) was converted to 1-chloroisoquinoline N-oxide using 0 potassium nitrite and conc HCl according to the general method described in the literature (Brown, E V J. Amer. Chem. Soc 1957, 79, 3565-3566) The title compound was prepared as a light brown solid MS (ES) m/e 179 9 (M + H)+ <br><br> b) l-[(3-Hydroxy-l-propyl)amino]-isoquinoline N-oxide <br><br> According to the procedure of Preparation 1 (a), except substituting 1-chloroisoqumoline N-oxide for the 2-chloropyridine-N-oxide hydrochloride, the title compound was prepared as an amber solid- MS (ES) m/e 219.1 (M + H)+ <br><br> Preparation 9 <br><br> Preparation of 2-IN-(3-methanesulfonvloxv-1 -propylVN-ftert-butoxvcarbonvllaminolDvridine-N-oxide a) 2-[N-(3-Hydroxy-l-propyl)-N-(ferr-butoxycarbonyl)amino]pyridine-N-oxide <br><br> A solution of 2-[(3-hydroxy-l-propyl)amino]pyridine-N-oxide (8 0 g, 47 6 mmol) in rm-BuOH (80 mL) was treated with di-terf-butyl dicarbonate (11 4 g, 55 3 mmol) After 18h, the solution was concentrated and the residue was triturated with hexane The <br><br> -38- <br><br> Printed from Mimosa <br><br> WO 99/15508 PCT/US98/19466 <br><br> resulting solid was dried in vacuo to give the title compound (12.5 g, 98%) as an off-white solid MS (ES) m/e 269 3 (M + H)+. <br><br> b) 2-[N-(3-Methanesulfonyloxy-1 -propyl)-N-(rm-butoxycarbonyl)amino]pyridine-N-5 oxide <br><br> Methanesulfonyl chloride (0.17 mL, 2 20 mmole) was added dropwise to a solution of 2-[N-(3-hydroxy-l-propyl)-N-(rm-butoxycarbonyl)amino]pyridine-N-oxide (0.50 g, 1 86 mmole) and pyridine (0 23 mL, 2 84 mmole) in CHCI3 (5 mL, dried over K2CO3) at 0°C When complete by TLC, the reaction was diluted with CHCI3, washed with ice water, 0 dried (Na2SC&gt;4), and concentrated. Silica gel chromatography (10% MeOH/CHCl3) gave the title compound (0 41 g, 64%) as a colorless oil. 'H NMR (250 MHz, CDCI3) 8 8 25 (dd, J = 6 0,1.9 Hz, 1 H), 7.25 (m, 4 H), 4 35 (t, J = 6 2 Hz, 2 H), 3.75 (t, I = 6 6 Hz, 2 H), 3 00 (s, 3 H), 2 00 (m, 2 H), 1 40 (s, 9 H). Unchanged 2-[N-(3-hydroxy-1-propyl)-N-(?ert-butoxycarbonyl)amino]pyridine-N-oxide (0 18 g, 36%) could also be recovered from the 5 chromatographic purification <br><br> Preparation 10 <br><br> Preparation of ethvl (±1-10.1 l-dihvdro-3-hvdroxv-5H-dibenzora.dkvcloheptene-10-acetate a) 6-Methoxy-l-phenylindene <br><br> A solution of 3.0 M phenylmagnesium bromide in Et20 (680 mL, 2 04 mole) under argon at ambient temperature was diluted with Et20 (700 mL) with stirring, and a solution of 6-methoxy-l-indanone (277 g, 1 71 mole) in THF (1400 mL) was added dropwise over 1 hr The reaction mixture was stirred for 2 h at ambient temperature and then was poured with stirring into saturated NH4CI (2 8 L) H2O (1.4 L) was added, and the organic phase separated The aqueous phase was extracted with Et20 (2 x 1 L), and the combined organic extracts were concentrated to give crude 6-methoxy-l-phenyl-1-indanol (445 g) as a brown oil. This oil was dissolved in toluene (2.5 L), and p-toluenesulfonic acid monohydrate (12 3 g, 0.065 mole) was added The solution was stirred and heated at reflux for 16 hr using a Dean-Stark trap with a condenser. H2O collection was minimal after 2 h and totaled 28 mL The solution was cooled and extracted sequentially with 5 % aqueous Na2C03 (1 L) and H2O (2 x 1 L) The organic layer was concentrated to give a dark brown oil (400 g). This oil was distilled under vacuum to give the title compound (298 2 g, 79%) as a yellow oil bp 152 - 190°C/2 0 Torr, TLC (10% EtOAc/hexanes) Rf 0.75. <br><br> -39- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> b) 2-Benzoyl-4-methoxyphenylacetic acid <br><br> Acetone (4 2 L) was chilled to 10°C, and a solution of 6-methoxy-l-phenyhndene (271 g, 1 22 mole) in acetone (1 8 L) was added over 1 5 hr concurrently with Jones reagent (1.8 L, prepared from CrC&gt;3 (470 g, 4 70 mole), H2O (1 L), and conc H2SO4 (405 mL)) 4 5 % Aqueous OSO4 (153 mL) was added to the resulting mixture in two portions, one at the onset of addition and the second at the mid-point of the addition, maintaining the temperature of the reaction mixture below 15°C Following the addition, the reaction mixture was warmed to 22°C and stirred for 1 5 h, during which time a mild exotherm increased the temperature to 28°C The reaction mixture was then cooled to below 20°C 0 and isopropanol (1 L) was added, dropwise initially and rapidly after the initial exotherm diminished Stirring became difficult during this phase The temperature reached 32°C during the isopropanol addition H2O (2 L) was added and the mixture was transferred to a separatory funnel Additional H2O was added to dissolve the precipitated chromous acid, and the mixture was extracted with CH2CI2 (2 L). The organic (upper) layer was separated 5 and the aqueous phase was extracted with CH2CI2 (2 x 1 L) The combined CH2CI2 <br><br> extracts were washed sequentially with H2O (2 L) and saturated brine (2 L), and then were concentrated to give a moist gray solid (416 g). This was triturated with a mixture of acetone and EtOAc and filtered and dried to give the title compound (225 4 g, 71%) as an off-white solid: mp 158-159°C <br><br> 0 <br><br> c) 2-Benzyl-4-methoxyphenylacetic acid <br><br> 2-Benzoyl-4-methoxyphenylacetic acid (215 5 g, 0 80 mole) was divided into two equal portions, and each was dissolved in glacial AcOH (1 5 L) in a 2.5 L pressure bottle. 5 % Pd/C (10 g, 0 0048 mole) was added to each, and each mixture was shaken at ambient :5 temperature under hydrogen on a Parr apparatus. After 2.5 hr, the mixtures were filtered to remove the catalyst, and the filter pads were washed with EtOAc The combined filtrates were concentrated to give the title compound (215 g, quantitative) as a heavy yellow oil which crystallized on standing 1H NMR (250 MHz, CDCI3) 8 7.05 - 7 35 (m, 6 H), 6.77 (dd, J = 8 3,2 7 Hz, 1 H), 6.71 (d, J = 2 7 Hz, 1 H), 4 00 (s, 2 H), 3.76 (s, 3 H), 3 54 (s, 2 10 H) <br><br> d) 10,1 l-Dihydro-3-methoxy-5H-dibenzo[a,d]cyclohepten-10-one <br><br> A solution of 2-benzyl-4-methoxyphenylacetic acid (215 g of crude material that contained 204 6 g (0 80 mole) of pure material) in CH2CI2 (1 L) was stirred under argon at 15 ambient temperature, and DMF (1 mL) was added, followed by oxalyl chloride (400 mL, 4 59 mole) The oxalyl chloride was added over 1 hr, dropwise initially to control the vigorous gas evolution The solution was stirred for 16 h at ambient temperature and then <br><br> -40- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> was concentrated to give the crude acid chloride (207 7 g, 0.756 mol, 95 %) as a yellow liquid This liquid was dissolved in CH2CI2 to a total volume of 500 mL, and the solution and AICI3 (100 8 g, 0.756 mol) were added concurrently over 1 hr to CH2CI2 (3 7 L) with stirring under argon at ambient temperature. The temperature was 28°C at the completion 5 of the addition The reaction mixture was stirred for 16 h at ambient temperature, during which time a solid precipitated H20 (1 L) was added, initially dropwise, over a period of 30 mm. The mixture was then separated and the organic phase was washed sequentially with H2O (1 L) and 5 % aqueous NaHCC&gt;3 (1 L) The CH2CI2 solution was then concentrated to give a yellow solid (175 3 g). Recrystallization from EtOAc/hexane gave 0 the title compound (128 g, 71%). mp 107 - 109°C <br><br> e) Ethyl (±)-10,11 -dihydro-10-hydroxy-3-methoxy-5H-dibenzo[a,d]cycloheptene-10-acetate <br><br> A 1 0 M solution of lithium bis(tnmethylsilyl)amide in hexanes (1282 mL, 1 282 mole) was added to THF (4 0 L) at -70°C under argon, then EtOAc (146 mL, 1 49 mole) was added dropwise over 20 min The reaction mixture was allowed to stir for 15 min, then N,N,N',N-tetramethylethlylenediamine (378 mL, 2 5 mole) was added over 20 min The reaction mixture was stirred for 10 min, then a solution of 10,1 l-dihydro-3-methoxy-5H-dibenzo[a,d]cyclohepten-10-one (119.2 g, 0 50 mol) m anhydrous THF (1 26 L) was added dropwise over 40 mm The temperature was maintained below -65°C during all of these additions The reaction mixture was stirred for 20 min at -65 to -70°C and then was poured into saturated aqueous NH4CI (6.2 L) with vigorous stirring. The organic layer was separated and the aqueous phase was extracted with EtOAc (2 x 1 L) The combined organic extracts were washed with H2O (2 x 1 L) and then were concentrated to give a light brown oil (175 g) Thin-layer chromatography (20 % EtOAc/hexanes) showed Rf 0 5 major (desired product) and Rf 0 7 minor (recovered ketone) The crude product was chromatographed on silica gel (2 kg, 10 % EtOAc/hexanes) to afford the title compound (101 g, 61 %) as a yellow oil. *H NMR (250 MHz, CDCI3) 5 7 63 (d, J = 7 7 Hz, 1 H), 7 00 - 7.30 (m, 4 H), 6.80 (d, J = 2 6 Hz, 1 H), 6 69 (dd, J = 8 2, 2.6 Hz, 1 H), 3.95 - 4 35 (m, 2 H), 4.07 (s, 2 H), 3 76 (s, 3 H), 3 68 (s, 1 H), 3 64 (d, J = 14 2 Hz, 1 H), 3.35 (d, J = 14 2 Hz, 1 H), 2.79 (d, J = 16.0 Hz, 1 H), 2.66 (d, J = 16.0 Hz, 1 H), 1 22 (t, J = 7.2 Hz, 3 H) <br><br> f) Ethyl (+)-10,l l-dihydro-3-methoxy-5H-dibenzo[a,d]cycloheptene-10-acetate <br><br> Ethyl (±)-10,11 -dihydro-10-hydroxy-3-methoxy-5H-dibenzo[a,d]cycloheptene-10-acetate (101 g, 0.31 mole) was dissolved in glacial acetic acid (1 8 L) and 12 N HCl (28 5 mL, 0 34 mole) was added. The mixture was placed in a 2 5 L pressure bottle containing <br><br> -41 - <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> 5% Pd/C (20 g, 0 0094 mole), and the resulting mixture was shaken at 35°C under hydrogen on a Parr hydrogenation apparatus equipped with a jacket heater After 18 hr, the reaction was cooled to ambient temperature, and the catalyst was removed by filtration The filtrate was concentrated to give a light yellow oil (85 1 g). This was chromatographed on silica gel (2 kg, step-gradient with 5 % to 10 % EtOAc/hexanes) to afford the title compound (69.1 g, 72%) as an oil: NMR (250 MHz, CDCI3) 8 7 05 - 7.22 (m, 4 H), 7 01 (d, J = 8.2 Hz, 1 H), 6 76 (d, J = 2 7 Hz, 1 H), 6 67 (dd, J = 8.2, 2 7 Hz, 1 H), 4 30 (d, J = 15 0 Hz, 1 H), 4 11 - 4.25 (m, 2 H), 3.85 (d, J = 15 0 Hz, 1 H), 3 70 - 3 90 (m, 1 H), 3 77 (s, 3 H), 3.31 (dd, J = 15.0,4.1 Hz, 1 H), 2.93 (dd, J = 15 0, 9.2 Hz, 1 H), 2 64 (dd, J = 15 6, 5 0 Hz, 1 H), 2 52 (dd, J = 15 6, 9 3 Hz, 1 H), 1 27 (t, J = 7.1 Hz, 3 H) <br><br> g) Ethyl (±&gt;-10,1 l-dihydro-3-hydroxy-5H-dibenzo[a,d]cycloheptene-10-acetate <br><br> A solution of ethyl (±)-10,l l-dihydro-3-methoxy-5H-dibenzo[a,d]cycloheptene-10-acetate (8.5 g, 0 027 mole) in CH2CI2 (150 mL) was chilled to -10°C with stirring under argon Ethanethiol (10 7 mL, 0.144 mole) was added, followed by AICI3 (20 6 g, 0 154 mole) in two portions over 15 min. An exotherm increased the temperature to 0°C following the additions, and the temperature was then increased to 25°C using a water bath. The reaction mixture was stirred at 25 to 30°C for 2 25 hr, at which point it was poured into ice-H20. The organic layer was separated, methanol (100 mL) was added, and the mixture was extracted with CH2CI2 (2 x 50 mL). The combined CH2CI2 extracts were washed with H2O (250 mL) and then were concentrated to give a viscous oil (8 6 g). This was taken up m Et20 (150 mL) and the ether was boiled off while replacing it with hexane The desired phenol first separated as an oil which crystallized on stirring at ambient temperature Two crops of solid were collected to afford the title compound (7 1 g, 89 %) mp 110- 112°C <br><br> Preparation 11 <br><br> HPLC separation of the enantiomers of ethvl C±V10.1 l-dihvdro-3-hvdroxv-5H-dibenzora.dlcvcloheptene-10-acetate a) Ethyl (R)-(+)-10,l l-dihydro-3-hydroxy-5H-dibenzo[a,d]cycloheptene-10-acetate and ethyl (S)-(-)-10,11 -dihydro-3-hydroxy-5H-dibenzo[a,d]cycloheptene-10-acetate <br><br> Ethyl (±)-10,1 l-dihydro-3-hydroxy-5H-dibenzo[a,d]cycloheptene-l 0-acetate was resolved into its enantiomers using the following conditions Daicel Chiralcel OJ® column (21 2 x 250 mm), 20% ethanol in hexane mobile phase, 15 mL/min flow rate, uv detection at 254 nm, 140 mg injection, tR for ethyl (S)-(-)-10,l l-dihydro-3-hydroxy-5H- <br><br> -42- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> dibenzo[a,d]cyclohexene-l0-acetate = 10 4 min , tg for ethyl (R)-(+)-10,l l-dihydro-3-hydroxy-5H-dibenzo[a,d]cyclohexene-10-acetate = 13 1 mm <br><br> Preparation 12 <br><br> Preparation of ethvl (±~)-10.1 l-dihvdro-7-fluoro-3-hvdroxv-5H-dibenzofa.dlcvcloheptene-10-acetate a) 1 -(3-Fluorophenyl)-6-methoxy-1 -indanol <br><br> According to the procedure of Preparation 10 (a), except substituting 3-fluorophenylmagnesium bromide for the phenylmagnesium bromide, the title compound was obtained as an amber oil MS (ES) m/e 276.0 (M + H)+ <br><br> b) 1 -(3-FluorophenyI)-6-methoxymdene <br><br> According to the procedure of Preparation 10 (a), except substituting l-(3-fluorophenyl)-6-methoxy-l-indanol for the 6-methoxy-l-phenyl-1-indanol, the title compound was obtained as a colorless oil following silica gel chromatography (4% EtOAc/hexanes). MS (ES) m/e 241.1 (M + H)+. <br><br> c) 2-(3-Fluorobenzoyl)-4-methoxyphenylacetic acid <br><br> According to the procedure of Preparation 10 (b), except substituting 2-(3-fluorophenyl)-6-methoxymdene for the 6-methoxy-l-phenylindene, the title compound was obtained as a white solid: MS (ES) m/e 289 2 (M + H)+ <br><br> d) 2-(3-Fluorobenzyl)-4-methoxyphenylacetic acid <br><br> According to the procedure of Preparation 10 (c), except substituting 2-(3-fluorobenzoyl)-4-methoxyphenylacetic acid for the 2-benzoyl-4-methoxyphenylacetic acid, the title compound was obtained as a colorless oil- MS (ES-) m/e 273 2 (M - H)" <br><br> e) 10,11 -Dihydro-7-fluoro-3-methoxy-5H-dibenzo[a,d]cyclohepten-10-one <br><br> According to the procedure of Preparation 10 (d), except substituting 2-(3-fluorobenzyl)-4-methoxyphenylacetic acid for the 2-benzyl-4-methoxyphenylacetic acid, the title compound was obtained as a white solid. Mp 129 - 130°C; MS (ES) m/e 279 2 (M + Na)+ <br><br> -43- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> f) Ethyl (±)-10,11 -dihydro-7-fluoro-10-hydroxy-3-methoxy-5H-dibenzo[a,d]cycloheptene-10-acetate <br><br> According to the procedure of Preparation 10 (e), except substituting 10,11-dihydro-7-fluoro-3-methoxy-5H-dibenzo[a,d]cyclohepten-10-one for the 10,11 -dihydro-3-methoxy-5H-dibenzo[a,d]cyclohepten-10-one, the title compound was obtained following silica gel chromatography (8% EtOAc/hexanes). MS (ES) m/e 362 2 (M + NH4)+. <br><br> g) Ethyl (±)-10,l l-dihydro-7-fluoro-3-methoxy-5H-dibenzo[a,d]cycloheptene-l 0-acetate <br><br> According to the procedure of Preparation 10 (f), except substituting ethyl (±)-10,ll-dihydro-7-fluoro-10-hydroxy-3-melhoxy-5H-dibenzo[a,d]cycloheptene-10-acetate for the ethyl (±)-10,l 1-dihydro-10-hydroxy-3-methoxy-5H-dibenzo[a,d]cycloheptene-10-acetate, the title compound was obtained as a colorless oil following silica gel chromatography (10% EtOAc/hexanes)- MS (ES) m/e 329 2 (M + H)+ <br><br> h) Ethyl (±)-10,l l-dihydro-7-fluoro-3-hydroxy-5H-dibenzo[a,d]cycloheptene-l 0-acetate <br><br> According to the procedure of Preparation 10 (g), except substituting ethyl (±)-10,1 l-dihydro-7-fluoro-3-methoxy-5H-dibenzo[a,d]cycloheptene-l 0-acetate for the ethyl (±)-10,l l-dihydro-3-methoxy-5H-dibenzo[a,d]cycloheptene-10-acetate, the title compound was obtained as a white solid following silica gel chromatography (1% MeOH/CH2Cl2) MS (ES) m/e 315 0 (M + H)+, 332 0 (M + NH4)+. <br><br> Preparation 13 <br><br> Preparation of ethvl (±V10.1 l-dihvdro-2-(,dimethvlamino')methvl-7-fluoro-3-hvdroxv-5H-dibenzof a.dlcvcloheptene-10-acetate a) Ethyl (±)-10,l l-dihydro-2-(dimethylamino)methyl-7-fluoro-3-hydroxy-5H-dibenzo[a,d]cycloheptene-10-acetate <br><br> To a solution of ethyl (±)-10,l l-dihydro-7-fluoro-3-hydroxy-5H-dibenzo [a,d]cycloheptene-10-acetate (0 4 g, 1 33 mmol) in 95% ethanol containing 2 M dimethylamine in MeOH (1.0 mL) was added 37% aqueous formaldehyde solution (0 5 mL) at RT under argon After 20 hr, the reaction was heated to reflux for 5 hr, then was concentrated on the rotavap The residue was partitioned between H2O and Et20, and the layers were separated The aqueous layer was extracted with Et20, and the combined organic layers were washed with brine, dried (MgSO,^), and concentrated on the rotavap to give the title compound (330 mg, 67%) as a colorless oil 'H NMR (400 MHz, CDCI3) 8 7 20 (m, 1 H), 6 88 (m, 2 H), 6.67 (s, 2 H),4 25 (d, J = 15 1 Hz, 1 H), 4 18 (q, 2 H), 3.78 <br><br> -44- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> (m, 1 H), 3 74 (d, J = 15 1 Hz, 1 H), 3.55 (s, 2 H), 3 20 (dd, 1 H), 2 80 (dd, 1 H). 2 60 (dd, 1 H), 2 53 (dd, 1 H), 2 29 (S, 6 H), 1 27 (t, 3 H), MS(ES) m/e 372 3 (M + H)+ <br><br> Preparation 14 <br><br> Preparation of ethvl (±1-10.1 l-dihvdro-3-hvdroxv-2-methvl-5H-dibenzofa.d'lcvcIoheptene-10-acetate a) Ethyl (±)-10,ll-dihydro-2-formyl-3-methoxy-5H-dibenzo[a,d]cycloheptene-10-acetate <br><br> POCI3 (17 mL) was added dropwise to a solution of ethyl (±)-10,l l-dihydro-3-methoxy-5H-dibenzo[a,d]cycloheptene-l0-acetate (1 0 g, 3 mmol) m dry DMF (40 mL) at RT under argon, and the dark solution was heated at 90°C for 48 hr The reaction was concentrated on the rotavap and the residue was partitioned between H2O and EtOAc The organic layer was separated, dried (MgS04), and concentrated on the rotavap The residue was reconcentrated from xylenes (to remove any remaining DMF) then was chromatographed on silica gel (7% EtOAc in hexanes) to afford the title compound (230 mg, 21 %) as a colorless oil MS (ES) m/e 339 3 (M + H)+ <br><br> b) Ethyl (±)-10,11 -dihydro-3-methoxy-2-methyl-5H-dibenzo[a,d]cycloheptene-10-acetate <br><br> A mixture of ethyl (±)-10,l l-dihydro-2-formyl-3-methoxy-5H-dibenzo[a,d]cycloheptene-l 0-acetate (220 mg, 0.65 mmol), 10% Pd/C (90 mg), glacial HO Ac (15 mL), and conc HCl (2 mL) was shaken at RT under hydrogen (60 psi). After 20 hr, the mixture was filtered through cehte®, and the filtrate was concentrated to afford the title compound (200 mg, 95%) as a colorless oil: MS (ES) m/e 325 2 (M + H)+. <br><br> c) Ethyl (±)-10,11 -dihydro-3-hydroxy-2-methyl-5H-dibenzo[a,d]cycloheptene-10-acetate <br><br> To dry CH2CI2 (30 mL) cooled in an ice bath was added diethylsulfide (0 38 mL, 3 3 mmol) followed by AICI3 (438 mg, 3 3 mmol) To this solution was added dropwise a solution of ethyl (±)-10,ll-dihydro-3-methoxy-2-methyl-5H-dibenzo[a,d]cycloheptene-10-acetate (200 mg, 0 6 mmole) in dry CH2CI2 (6 mL), and the resulting mixture was stirred at RT for 2 hr. The reaction was quenched with 1 0 N HCl (10 mL), and the layers were separated The organic layer was dried (MgS04) and concentrated on the rotavap to give the title compound (100 mg, 56%) as a colorless oil MS (ES) m/e 311 2 (M + H)+ <br><br> -45- <br><br> Printed from Mimosa <br><br> WO 99/15508 PCT/US98/19466 <br><br> Preparation 15 <br><br> Preparation of ethvl (+V10.1 l-dihvdro-3-hvdroxv-6-methvl-5H-dibenzofa.dlcvcloheptene-10-acetate <br><br> 5 <br><br> a) 6-Methoxy-1 -(2-methylphenyl)-1 -indanol <br><br> According to the procedure of Preparation 10 (a), except substituting 2-methylphenylmagnesium bromide for the phenylmagnesium bromide, the title compound was obtained as an oil. MS (ES) m/e 277 0 (M + Na)+ <br><br> 10 <br><br> b) 6-Methoxy-1 -(2-methylphenyl)indene <br><br> According to the procedure of Preparation 10 (a), except substituting 6-methoxy-l-(2-methylphenyl)-1 -indanol for the 6-methoxy-l-phenyl-1-indanol, the title compound was obtained as a colorless oil following silica gel chromatography (3% EtOAc/hexanes): MS 15 (ES) m/e 237.2 (M + H)+ <br><br> c) 4-Methoxy-2-(2-methylbenzoyl)phenylacetic acid <br><br> According to the procedure of Preparation 10 (b), except substituting 6-methoxy-l -(2-methylphenyl)indene for the 6-methoxy-l-phenyhndene, the title compound was 20 obtained as a viscous oil MS (ES) m/e 285 3 (M + Nlty)"1" <br><br> d) 4-Methoxy-2-(2-methylbenzyl)phenylacetic acid <br><br> According to the procedure of Preparation 10 (c), except substituting 4-methoxy-2-(2-methylbenzoyl)phenylacetic acid for the 2-benzoyl-4-methoxyphenylacetic acid, the title 25 compound was obtained as a viscous oil: MS (ES) m/e 288.2 (M + NH4)+. <br><br> e) 10,11 -Dihy dro-3-methoxy-6-methyl-5H-dibenzo[a,d]cyclohepten-10-one <br><br> According to the procedure of Preparation 10 (d), except substituting 4-methoxy-2-(2-methylbenzyl)phenylacetic acid for the 2-benzyl-4-methoxyphenylacetic acid, the title 30 compound was obtained as a white solid following silica gel chromatography (6% EtOAc/hexanes) MS (ES) m/e 253.0 (M + H)+. <br><br> f) Ethyl (±)-10,ll-dihydro-10-hydroxy-3-methoxy-6-methyl-5H-dibenzo[a,d]cycloheptene-10-acetate 35 According to the procedure of Preparation 10 (e), except substituting 10,11- <br><br> dihydro-3-methoxy-6-methyl-5H-dibenzo[a,d]cyclohepten-10-one for the 10,11 -dihydro-3- <br><br> -46- <br><br> Pnnted from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> methoxy-5H-dibenzo[a,d]cyclohepten-10-one, the title compound was obtained following silica gel chromatography (8% EtOAc/hexanes). MS (ES) m/e 358 2 (M + NH4)+ <br><br> g) Ethyl (±)-10,l l-dihydro-3-methoxy-6-methyl-5H-dibenzo[a,d]cycloheptene-l 0-acetate 5 According to the procedure of Preparation 10 (f), except substituting ethyl (±)- <br><br> 10,11 -dihydro-10-hydroxy-3-methoxy-6-methyl-5H-dibenzo[a,d]cycloheptene-10-acetate for the ethyl (±)-10,11 -dihydro- 10-hydroxy-3-methoxy-5H-dibenzo[a,d]cycloheptene-10-acetate, the title compound was obtained as a colorless oil following silica gel chromatography (5% EtOAc/hexanes) MS (ES) m/e 325 3 (M + H)+. <br><br> 10 <br><br> h) Ethyl (±)-10,l 1-dihydro-3-hydroxy-6-methyl-5H-dibenzo[a,d]cycloheptene-l 0-acetate <br><br> According to the procedure of Preparation 10 (g), except substituting ethyl (±)-10,1 l-dihydro-3-methoxy-6-methyl-5H-dibenzo[a,d]cycloheptene-l 0-acetate for the ethyl (±)-10,l l-dihydro-3-methoxy-5H-dibenzo[a,d]cycloheptene-l 0-acetate, the title compound 15 was obtained as a white solid after trituration with MeOH MS (ES) m/e 311 2 (M + H)+ <br><br> Preparation 16 <br><br> Preparation of ethvl (±&gt;-10.1 l-dihvdro-3-13-(4-nitro-l-oxopvndin-2-vlamino'&gt;-l-20 propvloxvl-5H-dibenzofa.d1cvcloheptene-10-acetate a) Ethyl (±)-10,l l-dihydro-3-[3-(4-nitro-l-oxopyndin-2-ylamino)-l-propyloxy]-5H-dibenzo [a,d]cycloheptene-10-acetate <br><br> A solution of 2-[(3-hydroxy-l-propyl)amino]-4-nitropyndine-N-oxide (0 85 g, 4 25 mmole) and diethyl azodicarboxylate (0.63 mL, 4 mmole) in anhydrous DMF (10 mL) was added dropwise to a solution of ethyl (±)-10,l l-dihydro-3-hydroxy-5H-dibenzo[a,d]cycloheptene-l 0-acetate (0 59 g, 2 mmole) and triphenylphosphine (1 10 g, 4 2 mmole) m anhydrous DMF (10 mL) at RT under argon After 23 hr, the reaction was concentrated and the residue was reconcentrated from xylenes (2 x). Silica gel 30 chromatography (gradient 1 1 EtOAc/hexanes, then EtOAc, then 5% MeOH in M <br><br> EtOAc/CHC^) gave crude title compound. Unchanged (±)-10,l l-dihydro-3-hydroxy-5H-dibenzo[a,d]cycloheptene-l 0-acetate could be recovered from the 1 1 EtOAc/hexanes fractions Rechromatography of the crude title compound (3% MeOH in 1 • 1 EtOAc/CHCl3) gave clean title compound (0.72 g, 73%) as a yellow foam. TLC (10% 35 MeOH m 1 1 EtOAc/CHCl3) Rf 0 59, !H NMR (250 MHz, CDC13) 5 8 19 (d, J = 7 1 Hz, 1 H), 7 46 (d, J = 2 9 Hz, 1 H), 7 35 (dd, J = 7.1,2 9 Hz, 1 H), 7.00 - 7 30 (m, 5 H), 7 00 (d, J = 8 2 Hz, 1 H), 6 81 (d, J = 2 6 Hz, 1 H), 6 70 (dd, J = 8 2,2 6 Hz, 1 H), 4 29 (d, J = 15 1 <br><br> -47- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/U S98/19466 <br><br> Hz, 1 H), 4.18 (q, J = 7.1 Hz, 2 H), 4.08 (t, J = 5.5 Hz, 2 H), 3 86 (d, J = 15.1 Hz, 1 H), 3 72 - 3 90 (m, 1 H), 3.59 (q, J = 6 3 Hz, 2 H), 3.30 (dd, J = 15 0, 4.2 Hz, 1 H), 2 93 (dd. J = 15.0, 9.3 Hz, 1 H), 2 64 (dd, J = 15 6,5.1 Hz, 1 H), 2 51 (dd, J = 15.6,9 3 Hz, 1 H), 2 10 - <br><br> 2 30 (m, 2 H), 1 27 (t, J = 7.1 Hz, 3 H); MS (ES) m/e 492 (M + H)+. <br><br> 5 <br><br> Preparation 17 <br><br> Preparation of ethvl (SV10.1 l-dihvdro-3-r3-(4-nitro-l-oxopvndin-2-vlaminoVl-propyloxv1-5H-dibenzofa.d1cvcloheptene-10-acetate <br><br> 10 <br><br> a) Ethyl (S)-10,1 l-dihydro-3-[3-(4-nitro-l-oxopyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate <br><br> According to the procedure of Preparation 16, except substituting ethyl (S)-10,l 1-dihydro-3-hydroxy-5H-dibenzo[a,d]cycloheptene-l 0-acetate for the ethyl (±)-10,11-15 dihydro-3-hydroxy-5H-dibenzo[a,d]cycloheptene-10-acetate, the title compound was prepared MS (ES) m/e 492 (M + H)+ <br><br> Preparation 18 <br><br> 20 Preparation of 10.1 l-Dihvdro-3-methoxv-5H-dibenzoFa.dlcvclohepten-10-one a) 2-Benzyl-4-methoxyphenylacetic acid <br><br> A solution of 2-benzoyl-4-methoxyphenylacetic acid (13 0 g, 0 048 mol), prepared by the method of J Med Chem 1981,24, 998, in glacial acetic acid (600 mL) was treated 25 under argon with 4 3 g. of 10% Pd/C and hydrogenated at 50 psi for 17 hours The mixture was filtered using cehte® and the filtrate was concentrated and reconcentrated from toluene and methylene chloride to give 14 2 of the title compound NMR (400 MHz, CDCI3) 8 <br><br> 3 52 (s, 2H), 3 75 (s, 3H), 4.0 (s, 3H), 6.7 (m, 2H), 7 15 (m, 6H). <br><br> 30 b) 10,ll-Dihydro-3-methoxy-5H-dibenzo[a,d]cyclohepten-10-one <br><br> A solution of 2-benzyl-4-methoxyphenylacetic acid (14.2 g, 0.055m) in benzene (120 mL) and thionyl chloride (28 mL) was refluxed for 1 hour and concentrated The acid chloride was dissolved in dry methylene chloride (40 mL), and the solution was added dropwise under argon to a solution of A1C13 (14 7 g, 0.11 mol) in methylene chloride (600 35 mL) The reaction was stirred under an argon atmosphere for 2 5 hours at room temperature, then was quenched with ice-water (200 mL) The layers were separated, and the organic phase was washed sequentially with 10% NaOH solution, water, and dil. HCl <br><br> -48- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> The resulting solution was diluted with ether (200 mL), dried over MgS04, and concentrated. The solid residue was triturated with ether/hexane (1 1) and 9 35 g of the title compound was collected by filtration: Mp 105 - 106°C; NMR (400 MHz, CDCI3) 8 3 72 (s, 3H), 4 1 (s, 2H), 4.2 (s, 2H), 6 7 (d, IH), 6.82 (s, IH), 7 30 (m, 4H), 8.1 (d, IH) <br><br> 5 <br><br> Preparation 19 <br><br> Preparation of ethvl (±)-10.1 l-dihvdro-3-methoxv-5H-dibenzora.d1cvcloheptene-l 0-acetate <br><br> 10 a) Ethyl (±) 3-(3-methoxyphenyl)indeneacetate <br><br> To a cold solution of 3-(3-methoxyphenyl)indene (4 g, 18 mmol), prepared by the method of J Med Chem 1981, 24, 998, in THF (15 mL) at 0°Cwas added dropwise a solution of LiN(TMS)2 (20 mL, 1M in THF) over 5 min. The resulting solution was added dropwise to a solution of ethyl bromoacetate (3.34 g, 20 mmol) in THF (15 mL) at -78°C 15 over 30 min After 2.5 h, the mixture was quenched with saturated ammonium chloride solution and the layers were separated The organic layer was dried over MgS04 and concentrated to give the crude product which was purified by column chromatography (Si02/2-4% EtOAc/hexane) to give title compound (1 1 g) *H NMR (400 MHz, CDCI3) 8 1 30 (t, 3H), 2 50 (m, IH), 2.85 (m, IH), 3 85 (s, 3H), 4 0 (m, IH), 4 20 (q, 2H), 6 6 (s, IH), 20 6 9 (m, IH), 7 2 (s, IH), 7 35 (m, 6H). <br><br> b) Ethyl (±) 3-[(3-methoxybenzoyl)]phenylsuccmate <br><br> A solution of ethyl (±) 3-(3-methoxyphenyl)mdeneacetate (1 1 g, 3.6 mmol) in acetone (30 mL) was treated with 4% aqueous solution of osmium tetroxide (0 5 mL) 25 followed by a dropwise addition of 1 2 M Jones reagent (5 mL, 6 mmol) according to the literature procedure (J Org Chem 1993, 58, 4745) After stirring overnight at room temperature, the dark reaction mixture was quenched with isopropanol (2 5 mL), followed by sodium bisulfite (0 9 g) and water (30 mL). The product was extracted with ethyl acetate, washed with brine, dried over MgSC&gt;4, and concentrated to give a solid residue 30 Trituration with 1.1 ether / hexane gave 0 76 g of the title compound 1H NMR (400 MHz, CDCI3) 8 1 18 (t, 3H), 2.90 (m, IH), 3.3 (m, IH), 3.92 (s, 3H), 4.1 (q, 2H), 4.4 (m, IH), 4.4 (d, IH), 7 25 (m, 2H), 7 5 (m, 6H). <br><br> c) Ethyl (±) 3-[(3-methoxybenzyl)]phenylsuccinate <br><br> 35 A mixture of ethyl (±) 3-[(3-methoxybenzoyl)]pheny [succinate (0.76 g., 2.1 mmol) <br><br> and 10% Pd/C (0 6 g) in glacial acetic acid (35 mL) was hydrogenated at 50 psi for 17 hours The mixture was filtered using celite® and the filter pad was washed with acetic <br><br> -49- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> acid. The filtrate was concentrated and reconcentrated from toluene and methylene chloride to give 0 65 g of the title compound 'H NMR (400 MHz, CDCI3) 8 1 20 (t, 3H), 2.20 (m, IH), 3 0 (m, IH), 3 74 (s, 3H), 4.1 (q, 2H), 4 18 (q, 2H), 4 4 (d, IH), 6 2 (m, 2H), 7.22 (m, 6H) <br><br> 5 <br><br> d) Ethyl (±)-10,l l-dihydro-3-methoxy-l l-oxo-5H-dibenzo[a,d]cycloheptene-10-acetate <br><br> To a magnetically stirred solution of ethyl (±) 3-[(3-methoxybenzyl)]phenylsuccmate (0.65 g, 1.9 mmol) in dry methylene chloride (10 mL) were added DMF (0 2 mL) and oxalyl chloride (0 2 mL, 2 28 mmol) After 1 5 h, the 10 solution was added dropwise to a suspension of aluminum chloride (0 6 g, 4 5 mmol) in dry methylene chloride (15 mL) The mixture was quenched after 2 h with ice water, the layers were separated, and the aqueous layer was extracted with methylene chloride. The combined organic layers were dried over MgSC&gt;4 and concentrated The residue was purified by column chromatography (Si02/2-4% EtOAc/hexane) to give title compound 15 (0.3 g): ]H NMR (400 MHz, CDCI3) 8 1.28 (t, 3H), 2.88 (m, IH), 3.55 (m, IH), 3.84 (s, 3H), 3 88 (d, IH), 4 18 (q, 2H), 4 85 (d, IH), 4 95 (m, IH), 5 8 (m, 2H), 7.22 (m, 4H), 8.1 (s, IH) <br><br> e) Ethyl (±)-10,l l-dihydro-3-methoxy-5H-dibenzo[a,d]cycloheptene-l 0-acetate <br><br> 20 A mixture of ethyl (±)-10,11 -dihydro-3-methoxy-11 -oxo-5H-dibenzo[a,d]cyc!oheptene-10-acetate (0 3 g , 0 93 mmol) and 10% Pd/C (0.3 g) in glacial acetic acid (25 mL) was hydrogenated at 50 psi for 18 hours The mixture was filtered using cehte® and washed with acetic acid The filtrate was concentrated and reconcentrated from toluene and methylene chloride to give 0 25 g of the title compound' 'H NMR (400 MHz, CDCI3) 8 25 1 28 (t, 3H), 2.60 (m, 2H), 2 90 (m, IH), 3.30 (m, IH), 3 80 (s, 3H), 3 85 (d, IH), 4 18 (q, 2H), 4.30 (d, IH), 6 70 (m, 2H), 7 0 (d, IH), 7 22 (m, 4H) <br><br> Preparation 20 <br><br> 30 Preparation of ethvl (±1-10.1 l-dihvdro-3-hvdroxv-dibenzorb.floxepine-10-acetate a) 4-Methoxy-2-phenoxyacetophenone <br><br> According to the procedure of Harris, T W. etal (J Med Chem 1982,25(7), 855 - 858), 2-fluoro-4-methoxyacetophenone (1 00 g, 5.95 mmole) was reacted with phenol to 35 give the title compound (1.27 g) as an oil. 'H NMR (300 MHz, CDCI3) 8 7 90 (d, J = 8.8 Hz, 1 H), 7 35 (m, 2 H), 7 20 (m, 1 H), 7 05 (m, 2 H), 6.70 (dd, J = 2 4, 8 8 Hz, 1 H), 6 35 (d, J = 2.4 Hz, 1 H), 3.75 (s, 3 H), 2.61 (s, 3 H). <br><br> -50- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> b) 2-(4-Methoxy-2-phenoxyphenyl)-1 -morphohn-4-ylethan-1 -thione <br><br> According to the procedure of Harris, T. W, et al. (J. Med. Chem 1982,25(7), 855 - 858), 4-methoxy-2-phenoxyacetophenone (1.69 g, 6 98 mmole), sulfur (0 36 g, 11.2 5 mmole), and morpholine (0 98 mL, 11 2 mmole) were reacted to give the title compound (1 24 g) as a white solid- MS (ES) m/e 344 0 (M + H)+ <br><br> c) 2-(4-Methoxy-2-phenoxyphenyI)acetic acid <br><br> To a solution of 2-(4-methoxy-2-phenoxyphenyl)-l-morphohn-4-ylethan-l-thione 0 (0.35 g, 1.02 mmole) in i-PrOH (15 mL) and H2O (15 mL) was added KOH (0.57 g, 10 2 mmole) The reaction was heated at reflux for 18 hr, then was cooled to RT, diluted with H2O, and washed with Et20. The aqueous layer was acidified to pH = 4 with conc. HCl and was extracted with CHCI3 The combined extracts were dried over MgSC&gt;4 and concentrated to give the title compound (0 22 g) as a white solid This was used without 5 further purification: MS (ES) m/e 259 0 (M + H)+ <br><br> d) 3-Methoxydibenzo[b,f]oxepin-10-one <br><br> A solution of 2-(4-methoxy-2-phenoxyphenyl)acetic acid (594 mg, 2.3 mmole) in thionyl chloride (10 mL) was heated at reflux for 30 mm, then was concentrated to dryness, 0 and the residue was reconcentrated from toluene The resulting residue was dissolved in dry CH2CI2 (3 mL), and the solution was added dropwise at RT to a suspension of AICI3 (673 mg, 5 06 mmole)in dry CH2CI2 (4 mL) in a flame-dried flask under argon After stirring for 2 5 hr, the mixture was diluted with CH2CI2 (10 mL) and washed sequentially with 1 0 N NaOH and brine Drying (MgS04), concentration, and flash chromatography on 5 silica gel (5% EtOAc/hexanes) gave the title compound (264 mg, 48%) as a light yellow oil- *H NMR (300 MHz, CDCI3) 5 3.80 (s, 3 H), 4 02 (s, 2 H), 6 74 - 8 08 (m, 7 H) <br><br> e) Ethyl (±)-10,l l-dihydro-10-hydroxy-3-methoxydibenzo[b,f]oxepine-10-acetate <br><br> Anhydrous EtOAc (0.94 mL, 9 6 mmole) was added dropwise to a solution of &gt;0 lithium bis(trimethylsilyl)amide (1.0 M in THF, 7 mL, 7 mmole) in dry THF (7 mL) in a flame-dried flask at -78 °C under argon. After 0.5 hr, TMEDA (2.4 mL, 16 mmole) was added After another 5 min, a solution of 3-methoxydibenzo[b,f]oxepin-10-one (760 mg, 3.2 mmole) in THF (2 mL) was added dropwise over 3 mm Additional dry THF (0 4 mL) was used in transfer The reaction was stirred at -78 °C to -40 °C for 1 hr, then was i5 quenched with saturated NH4CI (10 mL) The mixture was warmed to RT and extracted with EtOAc Drying (MgS04), concentration, and flash chromatography on silica gel (10% EtOAc/hexanes) gave the title compound as a clear oil 'H NMR (300 MHz, CDCI3) 8 <br><br> -51 - <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT /US98/19466 <br><br> 1 14 - 1.20 (t, 3 H), 1 21 - 1 30 (m, 1 H), 2 62 - 2 68 (dd, 1 H), 2 94 - 3 02 (dd, 1 H), 3 24 -3 30 (dd, 1 H), 3.40-3 46 (dd, 1 H), 3 40 - 3.46 (dd, 1 H), 3 78 (s, 3 H), 4 08 - 4 18 (m, 2 H), 6 60 - 7 26 (m, 6 H), 7.64 - 7.68 (dd, 1 H) <br><br> 5 f) Ethyl (±)-10,l l-dihydro-3-methoxydibenzo[b,f]oxepine-l 0-acetate <br><br> Boron trifluoride etherate (0.48 mL, 3 9 mmole) was added to a solution of ethyl (±)-10,l 1-dihydro-10-hydroxy-3-methoxydibenzo[b,f]oxepine-l 0-acetate (690 mg, 1 95 mmole) and tnethylsilane (0 62 mL, 3.9 mmole) in dry CH2CI2 at 0 °C under argon After 20 mm, the reaction was quenched with 5% NaHCC&gt;3, and the mixture was extracted with 10 CH2CI2. Drying (MgS04) and concentration gave a yellow oil. This was dissolved in absolute ethanol (20 mL) and 10 % Pd/C (413 mg, 0 39 mmole) was added The mixture was hydrogenated for 3 hr at 50 psi on a Parr hydrogenation apparatus The catalyst was removed by filtration through cehte®, and the filtrate was concentrated to afford the title compound (523 mg, 86%) as a clear oil 'H NMR (300 MHz, CDCI3) 8 7 18-6.58 (m, 7 15 H), 4.18 - 4 08 (m, 2 H), 3 80 (s, 3 H), 3.80 - 3.74(m, 1 H), 3.40 - 3 30 (dd, 1 H), 2 98 - 2 84 (dd, 1 H), 2 74 - 2 62 (dd, 1 H), 2 60 - 2 52(m, 1 H), 1 32 - 1 20 (t, 3 H) <br><br> g) Ethyl (±)-10,l l-dihydro-3-hydroxydibenzo[b,f]oxepine-l 0-acetate <br><br> A solution of ethyl (±)-10,l l-dihydro-3-methoxydibenzo[b,f]oxepine-10-acetate 20 (523 mg, 1 68 mmole) in CH2CI2 (6 8 mL) was added dropwise to a cold solution of BBr3 m CH2CI2 (1 0 M, 6 7 mL, 6.7 mmole) at 0 °C under argon The reaction was stirred for 20 min, then CH3OH (7 mL) was added carefully The mixture was concentrated and the residue was flash chromatographed on silica gel (15 - 20% EtOAc/hexanes) to afford the title compound (407 mg, 89%) as a pale yellow oil MS (ES) m/e 299 (M + H)+ <br><br> 25 <br><br> Preparation 21 <br><br> Preparation of 2-IY3-bromo-l-propvl')aminol-4-methvlpvridine-N-oxide hvdrobromide <br><br> 30 a) 2-[(3-Bromo-l-propyl)amino]-4-methylpyndine-N-oxide hydrobromide <br><br> A solution of SOBr2 (5.0 mL, 64.5 mmole) in CH2CI2 (20 mL) was added dropwise over 15-20 min to a solution of 2-[(3-hydroxy-l-propyl)ammo]-4-methylpyridine-N-oxide (10.0 g, 54.87 mmole) in CH2CI2 (100 mL) at 0 °C. The reaction was warmed to RT and stirred for 2 hr, then Et20 (200 mL) was added slowly The 35 solvents were decanted away from the gummy precipitate, and the precipitate was washed with additional CH2Cl2/Et20 (several times). The resulting brownish-yellow residue solidified on standing in a refrigerator overnight. This solid was collected and washed with <br><br> -52- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> Et20 to afford the title compound (15.07 g) as a yellow solid. Additional title compound (2 05 g) was obtained as white needles by concentration of the combined organic layers The total yield of title compound was 17-89 g (96%) MS (ES) m/e 245 and 247 (M + H)+ <br><br> 5 The following compounds illustrate methods for preparing the biologically active compounds of this invention from intermediate compounds such as described in the foregoing Preparations <br><br> Example 1 <br><br> 0 <br><br> Preparation of (+V10.1 l-dihvdro-3-f4-Cpvridin-2-vlamino)-l-butvn-5H-dibenzora.dlcvcloheptene-10-acetic acid a) 4-(2-Tetrahydropyranyloxy)-1 -tributylstannyl-1 -butyne <br><br> A solution of n-butyllithium in hexanes (1.6 M, 18.8 mL, 30 mmole) was added in a stream over 2 mm to a solution of 2-(3-butynyloxy)tetrahydro-2//-pyran (4 7 mL, 30 mmole) m dry THF (60 mL) at 0°C under argon After 0 5 hr, tnbutyltin chloride (8 1 mL, 30 mmole) was added all at once, and the reaction was warmed to RT After 3 hr, the reaction was diluted with hexanes (300 mL) and washed sequentially with H2O (2 x 60 mL), 10% KF (2 x 30 mL), and saturated brine (60 mL) Drying (Na2SC&gt;4), concentration, and silica gel chromatography (3% EtOAc/hexanes) gave the title compound (3 58 g, 27%) as a nearly colorless oil. TLC (5% EtOAc/hexanes) Rf 0 37; ^H NMR (400 MHz, CDCI3) 6 4 66 (narrow t, 1 H),375-396 (m,2H),349-362(m,2H),256 (appt,2H), 1 76-1.91 (m, 1 H), 1.65 - 1 78 (m, 1 H), 1.42- 1 65 (m, 10 H), 1 22- 1 41 (m,6H),0 82- 1 08 (m, 15 H) <br><br> b) Ethyl (±)-10,l l-dihydro-3-[4-(2-tetrahydropyranyloxy)-l-butyn-l-yl]-5H-dibenzo[a,d]cycloheptene-10-acetate <br><br> A mixture of ethyl (±)-10,l l-dihydro-3-(tnfluoromethanesulfonyloxy)-5H-dibenzo[a,d]cycloheptene-l0-acetate (1 34 g, 3.13 mmole), 4-(2-tetrahydropyranyloxy)-l-tnbutylstannyl-1-butyne (1.66 g, 3.76 mmole), LiCl (398 mg, 9.39 mmole), bis(tnphenylphosphine)palladium dichlonde (110 mg, 0 094 mmole), and anhydrous dioxane (31 mL) was heated at reflux under argon After 1 5 hr, the reaction was concentrated to remove most of the dioxane, and the residue was taken up in Et20 (100 mL) 10% KF (50 mL) was added and the mixture was stirred briskly for 0.5 hr The aqueous layer was removed and the Et20 layer was filtered through a mixture of cehte® and MgS04 The filtrate was concentrated and the residue was chromatographed on silica <br><br> -53- <br><br> Pnnted from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> gel (10% EtOAc/hexanes) to afford the title compound (1 12 g, 83%) as a pale yellow oil TLC (20% EtOAc/hexanes) Rf 0.40, NMR (400 MHz, CDCI3) 5 7 21 - 7.30 (m. 1 H), 7 06 -7 20 (m, 5 H), 7.00 (d, J = 7 8 Hz, 1 H), 4 69 (t, J = 3.6 Hz, 1 H), 4 31 (d, J = 15.2 Hz, <br><br> 1 H), 4 11 - 4.23 (m, 2 H), 3 76 - 3 97 (m, 4 H), 3 59 - 3 68 (m, 1 H), 3 48 - 3 57 (m. 1 H), 5 3 34 (dd, J = 15 2, 4 1 Hz, 1 H), 2 97 (dd, J = 15 2, 9.5 Hz, 1 H), 2 70 (t, J = 7 3 Hz. 2 H), <br><br> 2 65 (dd, J = 15 7,4 8 Hz, 1 H), 2 51 (dd, J — 15 7, 9.5 Hz, 1 H), 1.78 - 1.92 (m, 1 H), 1 68 -1.78 (m, 1 H), 1 44 - 1.68 (m, 4 H), 1.27 (t, J = 7.1 Hz, 3 H); MS (ES) m/e 455 (M + Na)+. <br><br> c) Ethyl (±)-10,l l-dihydro-3-[4-(2-tetrahydropyranyloxy)-l-butyl]-5H-0 dibenzo[a,d]cycloheptene-10-acetate <br><br> A mixture of ethyl (±)-10,l l-dihydro-3-[4-(2-tetrahydropyranyloxy)-l-butyn-l-yl]-5H-dibenzo[a,d]cycloheptene-l 0-acetate (1 2 g, 2 77 mmole), 10% Pd/C (0.3 g, 0 28 mmole), and EtOAc (28 mL) was shaken at RT under hydrogen (50 psi) on a Pan-apparatus After 3 hr, the reaction was filtered through cehte® and the filtrate was 5 concentrated Silica gel chromatography (10% EtOAc/hexanes) gave the title compound (1 06 g, 88%) as a colorless oil. TLC (20% EtOAc/hexanes) Rf 0.51, *H NMR (400 MHz, CDCI3) 8 7.05 - 7.20 (m, 4 H), 6.92 - 7.03 (m, 3 H), 4.53 - 4 60 (m, 1 H), 4.34 (d, J = 15.1 Hz, 1 H), 4 12 - 4.26 (m, 2 H), 3 80 - 3 90 (m, 3 H), 3 71 - 3 80 (m, 1 H), 3.44 - 3 53 (m, 1 H), 3 35 - 3 44 (m, 1 H), 3 33 (dd, J = 15.1,4.1 Hz, 1 H), 2 95 (dd, J = 15 1, 9 4Hz, 1 H), 0 2.65 (dd, J = 15.5,4 9 Hz, 1 H), 249 - 2 61 (m, 3 H), 1.77- 1 90 (m, 1 H), 1 45 - 1 77 (m, 9 H), 1 27 (t, J = 7 1 Hz, 3 H); MS (ES) m/e 459 (M + Na)+ <br><br> d) Ethyl (±)-10,l l-dihydro-3-(4-hydroxy-l-butyl)-5H-dibenzo[a,d]cycloheptene-10-acetate <br><br> :5 A solution of ethyl (±)-10,11 -dihydro-3-[4-(2-tetrahydropyranyloxy)-1 -buty 1]-5H- <br><br> dibenzo[a,d]cycloheptene-l 0-acetate (456 0 mg, 1 04 mmole) and /j-toluenesulfonic acid monohydrate (60 mg, 0 31 mmole) in absolute EtOH (10 mL) was stirred at RT After 2 hr, the reaction was quenched with 5 % NaHC03 (1 mL) and concentrated to remove the EtOH. The residue was diluted with H2O (2 mL) and extracted with CH2CI2 Drying 0 (MgS04), concentration, and silica gel chromatography (1 1 EtOAc/hexanes) gave the title compound (342.4 mg, 93%) as a colorless oil. TLC (1.1 EtOAc/hexanes) Rf 0 49; *H NMR (250 MHz, CDCI3) 8 6 85 - 7.25 (m, 7 H), 4 34 (d, J = 15 1 Hz, 1 H), 4 08 - 4 30 (m, 2 H), 3 75 - 3.95 (m, 2 H), 3.53 - 3 72 (m, 2 H), 3.33 (dd, J = 15 1,4 1 Hz, 1 H), 2 95 (dd, J = 15.1, 9 4 Hz, 1 H), 2 40 - 2 75 (m, 4 H), 1 45 - 1 80 (m, 4 H), 1 27 (t, J = 7 1 Hz, 3 H), (5 MS (ES) m/e 353 (M + H)+ <br><br> -54- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/U S98/19466 <br><br> e) Ethyl (±)-10,l l-dihydro-3-[4-(N-phthalidimido)-l-butyI]-5H-dibenzo[a,d]cycloheptene-10-acetate <br><br> Diethyl azodicarboxylate (0 2 mL, 1 26 mmole) was added dropwise to a solution of ethyl (±)-10,l ]-dihydro-3-(4-hydroxy-l-butyl)-5H-dibenzo[a,d]cycloheptene-l 0-acetate 5 (0 37 g, 1 05 mmole), tnphenylphosphme (0 33 g, 1 26 mmole), and phthalimide (0 19 g, 1 26 mmole) in anhydrous THF (10 mL) at RT under argon. After 23 hr, the reaction was concentrated on the rotavap Silica gel chromatography (30% EtOAc/hexanes) gave the title compound (0.35 g, 70 %) as a colorless oil- MS (ES) m/e 504 3 (M + Na)+ <br><br> 0 f) Ethyl (±)-10,11 -dihydro-3-(4-amino-1 -butyl)-5H-dibenzo[a,d]cycloheptene-10-acetate Hydrazine monohydrate (0 11 g, 2.18 mmole) was added to a solution of ethyl (±)-10,1 l-dihydro-3-[4-(N-phthahdimido)-l-butyl]-5H-dibenzo[a,d]cycloheptene-10-acetate (0 35 g, 0 73 mmole) in absolute EtOH (10 mL) and toluene (2 mL) at RT. The reaction was stirred at RT for 17 hr, then was filtered, and the filter pad was washed with toluene 5 Concentration on the rotavap gave the title compound (0 23 g, 90%) as a colorless solid MS (ES) m/e 352.3 (M + H)+ <br><br> g) Ethyl (+)-10,l l-dihydro-3-[4-(l-oxopyridin-2-ylamino)-l-butyl]-5H-dibenzo[a,d]cycloheptene-10-acetate <br><br> 0 A mixture of 2-chloropyndme-N-oxide hydrochloride (0 31 g, 1.88 mmole), ethyl <br><br> (±)-10,l l-dihydro-3-(4-amino-l-butyl)-5H-dibenzo[a,d]cycloheptene-l 0-acetate (0.22 g, 0 63 mmole), and NaHC03 (0.26 g, 3 13 mmole) in tert-amyl alcohol (6 mL) was heated at reflux for 21 hr. The reaction mixture was diluted with CH2CI2 (100 mL) and filtered, and the filtrate was concentrated on the rotavap Silica gel chromatography (1 9.5 15 MeOH/CH2Cl2/EtOAc) gave the title compound (82 mg, 30%) as a yellow oil MS (ES) m/e 445 2 (M + H)+. <br><br> h) Ethyl (±)-10,l l-dihydro-3-[4-(pyridin-2-ylamino)-l-butyl]-5H-dibenzo[a,d]cycloheptene-10-acetate <br><br> 10 A mixture of ethyl (±)-10,11 -dihydro-3-[4-( 1 -oxopyridin-2-ylamino)-1 -butyl]-5H- <br><br> dibenzo[a,d]cycloheptene-l 0-acetate (0 07 g, 0.16 mmole), 10% Pd/C (0.08 g, 0.075 mmole), cyclohexene (0.16 mL, 1.6 mmole), and isopropanol (4 mL) was heated at reflux under argon for 14 hr, then the catalyst was removed by filtration through cehte® The filter pad was washed with isopropanol and MeOH, and the filtrate was concentrated on the 15 rotavap to give the title compound (0 046g, 69%) as a clear oil. MS (ES) m/e 429 3 (M + h)+ <br><br> -55- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> 1) Ethyl (±)-10,l l-dihydro-3-[4-(pyridin-2-ylamino)-l-butyl]-5H-dibenzo[a,d]cycloheptene-10-acetic acid <br><br> A mixture of ethyl (±)-10,l l-dihydro-3-[4-(pyridin-2-ylamino)-l-butyl]-5H-dibenzo[a,d]cycloheptene-l 0-acetate (46 mg, 0.11 mmole) and 1 0 N LiOH (0 66 mL, 0 66 5 mmole) in THF (3 mL) and H2O (3 mL) was stirred at RT After 24 hr, the reaction mixture was concentrated on the rotavap, and the residue was diluted with H2O (5 mL). The solution was cooled in an ice bath, and ION AcOH was added slowly to give a white precipitate. Chromatography on C-18 YMC (45% CH3CN/H2O containing 0.1% TFA) gave the title compound (13 mg, 21%) as a white solid: MS (ES) m/e 401 3 (M + H)+. 10 Anal. Calcd for C26H28N2O2 0 75 H20 • 1.5 CF3C02H. C, 59.54; H, 5.31, N, 4 72 Found C, 59 69, H, 5.31; N, 4.72 <br><br> Example 2 <br><br> 15 Preparation of (±)-10.1 l-dihvdro-3-r3-(4-ethoxvpvridin-2-vlamino)-l-Dropvloxvl-5H-dibenzora.dkvcloheptene-10-acetic acid a) Ethyl (+)-10,l l-dihydro-3-[3-(4-ethoxy-l-oxopyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate 20 Ethyl (±)-10,l l-dihydro-3-[3-(4-nitro-l-oxopyridin-2-ylamino)-l-propyloxy]-5H- <br><br> dibenzo[a,d]cycloheptene-10-acetate (0.67 g, 1.36 mmole), 1.0 M NaOEt in ethanol (6 8 mL, 6 8 mmole), and absolute ethanol (6 8 mL) were combined, and the mixture was warmed in an oil bath preset at 70°C. A dark solution was produced, which was warmed for 10 min, then the oil bath was removed, and the solution was allowed to stir for an 25 additional 5-7 min without external heating The resulting solution was cooled in ice, and the reaction was quenched with glacial acetic acid (0 47 mL, 8 2 mmole). The mixture was concentrated and the residue was partitioned between CH2CI2 (10 mL) and half-saturated NH4CI (10 mL) The layers were separated and the aqueous layer was extracted with CH2CI2 (2x10 mL) The combined organic layers were dried (MgS04) and concentrated, 30 and the residue was reconcentrated from toluene to leave a reddish-orange oil Silica gel chromatography (5% MeOH/CHClj) gave the title compound (601.1 mg, 90%) as a yellow oil TLC (5% MeOH/CHCl3) Rf 0.36; JH NMR (250 MHz, CDCI3) 8 7 95 (d, J = 7.1 Hz, 1 H), 6 88 - 7.30 (m, 6 H), 6 77 (d, J = 2 6 Hz, 1 H), 6.67 (dd, J = 8.2, 2 6 Hz, 1 H), 5.95 -6 20 (m, 2 H), 4.28 (d, J = 15 0 Hz, 1 H), 4.18 (q, J = 7.2 Hz, 2 H), 4 04 (t, J = 5 6 Hz, 2 H), 35 3 65 - 4 00 (m, 4 H), 3 46 (q, J = 6 5 Hz, 2 H), 3 30 (dd, J = 15.0,4.2 Hz, 1 H), 2 93 (dd, J = 15 0,9 2 Hz, 1 H), 2 65 (dd, J = 15 6,5.0 Hz, 1 H), 2 52 (dd, J = 15 6,9 4 Hz, 1 H), 1 95-2.25 (m, 2 H), 1 10 - 1 45 (m, 6 H), MS (ES) m/e 491 (M + H)+ <br><br> -56- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT /US98/19466 <br><br> b) Ethyl (±)-10,l l-dihydro-3-[3-(4-ethoxypyridin-2-ylammo)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate <br><br> A mixture of ethyl (±)-10,ll-dihydro-3-[3-(4-ethoxy-l-oxopyridin-2-ylamino)-l-5 propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate (601.1 mg, 1.23 mmole), cyclohexene (1 2 mL, 12 3 mmole), 10% Pd/C (130 mg, 0 012 mmole), and absolute ethanol (12.3 mL) was heated at reflux under argon After 23 5 hr, the reaction was hot-filtered through celite® and the filter pad was washed with ethanol The filtrate was concentrated and the residue was reconcentrated from toluene Silica gel chromatography (5% MeOH in 1 • 1 10 EtOAc/CHCl3) gave the title compound (528 1 mg, 90%) as a light yellow oil. TLC (10% MeOH in EtOAc/CHCl3) Rf 0 67; &gt;H NMR (400 MHz, CDC13) 8 7 89 (d, J = 5 8 Hz, 1 H), 7 05 - 7.18 (m, 4 H), 6.99 (d, J = 8 2 Hz, 1 H), 6 77 (d, I = 2 6 Hz, 1 H), 6.66 (dd, J = 8 2, 2 6 Hz, 1 H), 6 17 (dd, J = 5 8, 2 1 Hz, 1 H), 5.86 (d, J = 2 1 Hz, 1 H), 4 73 (brt, 1 H), 4.28 (d, J = 149Hz, 1 H),4 11 -4.25 (m,2H),4 04(t, J = 5 9Hz, 2H), 3 98 (q, J = 70Hz,2 15 H), 3 83 (d, J = 14 9 Hz, 1 H), 3 76 - 3 85 (m, 1 H), 3 43 (q, J = 6 4 Hz, 2 H), 3.30 (dd, J = 15 0,4.1 Hz, 1 H), 2 93 (dd, J = 15 0,9 2 Hz, 1 H), 2.64 (dd, J = 15 6,4 8 Hz, 1 H), 2.52 (dd, J = 15.6, 9 5 Hz, 1 H), 2.01 -2 11 (m,2H), 1 37 (t, J = 7.0 Hz, 3 H), 1 27 (t, J = 7 0 Hz, 3 H); MS (ES) m/e 475 (M + H)+ <br><br> 20 c) (+)-10,l l-Dihydro-3-[3-(4-ethoxypyridin-2-ylamino)-l-propyloxy]-5H-dibenzoIa,d]cycloheptene-10-acetic acid <br><br> 1 0 N NaOH (1 7 mL, 1 7 mmole) was added dropwise to a solution of ethyl (+)-10,11 -dihydro-3-[3-(4-ethoxypyridin-2-ylamino)-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene-l 0-acetate (528.1 mg, 1 11 mmole) in absolute ethanol (11 mL) 25 at RT, and the solution was warmed in an oil bath preset at 45°C. After 20 hr, the reaction was concentrated, and the residue was reconcentrated from H2O. The resulting residue was dissolved in H2O (10 mL) and the solution was filtered The pH was adjusted to 7 with 1.0 N HCl, and the mixture was stirred briskly to convert the initially-formed gummy precipitate into a solid. Trituration with a glass rod and a spatula aided in this 30 transformation The pH of the resulting mixture was readjusted to 7, and the solid was collected and washed with plenty of H2O. The filtrate was concentrated and the residue was dissolved m H2O with the aid of a little 1 0 N NaOH The pH was adjusted to 7 to afford a small second crop The crops were combined and dried in vacuum (40 - 50°C) to afford the title compound (453 7 mg, 82%) as an off-white solid HPLC (Hamilton PRP-35 1®, 45% CH3CN/H20 containing 0 1% TFA) k' = 1.32, 'h NMR (400 MHz, DMSO-d6) 8 7.78 (d, J = 6 6 Hz, 1 H), 7 35 - 7 65 (m, 1 H), 7.02 - 7.22 (m, 4 H), 6 97 (d, J = 8 3 Hz, 1 H), 6 82 (d, J = 2 4 Hz, 1 H), 6 68 (dd, J = 8 3, 2 4 Hz, 1 H), 6 29 (dd, 1 H), 6.15 (narrow d, <br><br> -57- <br><br> Pnnted from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> 1 H), 4 20 (d, J = 14.6 Hz, 1 H), 3 93 - 4 12 (m, 4 H), 3 89 (d, J = 14 6 Hz, 1 H), 3 60 - 3 71 (m, 1 H), 3 30 - 3 50 (m, 2 H), 3 20 (dd, J = 15.1,4 1 Hz, 1 H), 2 83 (dd, J = 15 1, 10 1 Hz, <br><br> 1 H), 2 60 (dd, J = 16.0, 5 3 Hz, 1 H), 2 48 (dd, J = 16 0, 8 9 Hz, 1 H, partially obscured by residual solvent signal), 1 90 - 2.05 (m, 2 H), 1 30 (t, J = 6.9 Hz, 3 H), MS (ES) m/e 447 (M <br><br> 5 + H)+ Anal Calcd for C27H30N2O4 • 1.5 HCl- C, 64 70, H, 6.33, N, 5 59. Found C, 64.53; H, 6 14; N, 5.31 <br><br> Example 3 <br><br> 0 Preparation of (±1-10.11-dihvdro-3-f2-f2-(ethvlamino)thiazol-4-vll-l-ethoxvl-5H-dibenzora.dlcvcloheptene-10-acetic acid a) Ethyl (±)-10,l l-dihydro-3-[2-[2-(ethylamino)thiazol-4-yl]-l-ethoxy]-5H-dibenzo[a,d]cycloheptene-10-acetate <br><br> 5 A solution of 2-(ethylammo)-4-thiazoleethanol (0.33 g, 1.9 mmole) and diethyl azodicarboxylate (0.30 mL, 1 9 mmole) in anhydrous DMF (5 mL) was added dropwise over 5 mm to a solution of ethyl (±)-10,l l-dihydro-3-hydroxy-5H-dibenzo[a,d]cycloheptene-l 0-acetate (296 4 mg, 1 mmole) and tnphenylphosphme (525 mg, 2 mmole) in anhydrous DMF (5 mL) at RT The reaction was kept cool in a RT water :0 bath during the addition After 16 hr, the reaction was concentrated and the residue was reconcentrated from xylenes (2 x) Silica gel chromatography (20% EtOAc/hexanes) gave the title compound (145 0 mg, 32%) as a yellow oil TLC (1 1 EtOAc/hexanes) Rf 0.60, 1H NMR (250 MHz, CDCI3) 5 7.00 - 7 30 (m, 4 H), 6 98 (d, J = 8.2 Hz, 1 H), 6 77 (d, J = <br><br> 2 6 Hz, 1 H), 6 68 (dd, J = 8 2,2 6 Hz, 1 H), 6 21 (s, 1 H), 5 00 - 5.25 (m, 1 H), 4 04 - 4 38 !5 (m, 5 H), 3.81 (d, J = 15 1 Hz, 1 H), 3 70 - 3 90 (m, 1 H), 3.13 - 3 40 (m, 3 H), 2.99 (t, J = <br><br> 6.7 Hz, 2 H), 2 92 (dd, J = 14 9, 9.3 Hz, 1 H), 2 64 (dd, J = 15 6, 5 0 Hz, 1 H), 2 51 (dd, J = 15 6, 9 3 Hz, 1 H), 1 27 (t, J = 7.2 Hz, 3 H), MS (ES) m/e 451 (M + H)+ <br><br> b) (±)-10,1 l-Dihydro-3-[2-[2-(ethylamino)thiazol-4-yl]-l-ethoxy]-5H-10 dibenzo[a,d]cycloheptene- 10-acetic acid <br><br> 1.0 N LiOH (0.32 mL, 0 32 mmole) was added dropwise to a solution of ethyl (±)-10,11 -dihydro-3-[2-[2-(ethylamino)thiazol-4-yl]-1 -ethoxy]-5H-dibenzo[a,d]cycloheptene-10-acetate (145 0 mg, 0 32 mmole) in THF (2 4 mL) and H2O (0 48 mL) at 0°C. The resulting pinkish-orange two-phase mixture was stirred at 0°C for 10 min, during which 15 time the color faded to orangish-yellow, then was warmed to RT After 1 5 hr, a little more H2O (5 drops) was added, and the reaction was stirred for 42 hr, then was cooled to 0°C and neutralized with TFA (0 025 mL). The THF was removed on the rotavap, and the <br><br> -58- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> resulting oily residue was diluted with 0.1% TFA/CH3CN to give a homogeneous solution ODS chromatography (gradient 40% CH3CN/H2O containing 0 1% TFA, then 45% CH3CN/H2O containing 0.1 % TFA) gave fractions containing the title compound These were pooled, and the CH3CN was removed on the rotavap. The resulting aqueous mixture 5 was made basic at 0°C to afford a homogeneous solution Careful acidification to pH 4 - 5 with 1 0 N HCl gave a solid precipitate, which was collected, washed with plenty of H2O, and dried to afford the title compound (80 9 mg, 51 %) as an off-white solid HPLC (Hamilton PRP-1®, 45% CH3CN/H20 containing 0 1% TFA) k' = 0.89, JH NMR (400 MHz, CD3OD) 8 7.02 - 7 18 (m, 4 H), 7 00 (d, J = 8 3 Hz, 1 H), 6.79 (d, J = 2 6 Hz, 1 H), 10 6.68 (dd, J = 8 3, 2.6 Hz, 1 H), 6 45 (s, 1 H), 4.26 (d, J = 14.9 Hz, 1 H), 4.20 (t, J = 6 4 Hz, 2 H), 3 87 (d, J = 14 9 Hz, 1 H), 3.68 - 3 80 (m, 1 H), 3 34 (q, J = 7 3 Hz, 2 H, partially obscured by residual solvent signal), 3 30 (dd, 1 H, obscured by residual solvent signal), 2 99 (t, J = 6 4 Hz, 2 H), 2 92 (dd, J = 15.0, 9.4 Hz, 1 H), 2 62 (dd, J = 15.9, 5 0 Hz, 1 H), 2 47 (dd, J = 15.9, 9.3 Hz, 1 H), 1 27 (t, J = 7 3 Hz, 3 H); MS (ES) m/e 423 (M + H)+. 15 Anal. Calcd for C24H26N2O3S 0.67 CF3C02H: C, 61 03, H, 5 39, N, 5 62 Found C, 61.21; H, 5.36; N, 5.60. <br><br> Example 4 <br><br> 20 Preparation of (S1-10.11 -dihvdro-3-r3-(pvridin-2-vlaminoV 1 -propvloxv1-5H-dibenzora.dlcvcloheptene-10-acetic acid a) Ethyl (S)-l 0,11 -dihydro-3-[3-( 1 -oxopyridin-2-ylamino)-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate 25 To a stirred solution of ethyl (S)-10,l l-dihydro-3-hydroxy-5H- <br><br> dibenzo[a,d]cycloheptene-10-acetate (35 g, 118 mmol) in dry THF (1 1 L) and dry DMF (600 mL) under argon were added 2-[(3-hydroxy-l-propyl)amino]pyridine-N-oxide (29 4 g, 175 mmol) and tnphenylphosphine (45.9 g, 175 mmol) After all solids had completely dissolved (-1 h), the reaction was cooled to 0 °C in an ice bath and dnsopropyl 30 azodicarboxylate (36 4 mL, 95%, 175 mmol) was added via syringe The reaction was allowed to warm slowly to RT and was stirred for 18 h Concentration and flash chromatography on silica gel (95:5 CHCl3/MeOH) followed by a second purification by flash chromatography on silica gel (80'20'5 CHC^/EtOAc/EtOH) gave the title compound (37 66 g, 71 %) as a pale yellow solid foam 1H NMR (400 MHz, DMSO-d6) 8 8 08 (dd, J 35 = 6 3,1 1 Hz, IH), 7.29 (t, IH), 7.19-7 06 (m, 5H), 6.97 (d, J = 8 3 Hz, IH), 6 84 (d, J = 2.5, IH), 6 79 (dd, J = 8 5, 1 6 Hz, IH), 6.69 (dd, J = 8 3, 2 6 Hz, IH), 6 57 (m, IH), 4 17 (d, J = 14 7 Hz, IH), 4 13-4 07 (m, 2H), 4 00 (t, 2H), 3 91 (d, J = 14.7 Hz, IH), 3 66 (m, <br><br> -59- <br><br> Printed from Mimosa <br><br> WO 99/15508 PCT/US98/19466 <br><br> IH), 3 39 (t, 2H), 3 19 (dd, J = 15.1,4 5 Hz, IH), 2 85 (dd, J = 15.1,10 0 Hz, IH). 2 65 (dd, J = 15 8, 5.4 Hz, IH), 1.99 (m, 2H), 1 18 (t, 3H), MS (ES) m/e 447 3 (M + H)+ <br><br> b) Ethyl (S)-10,1 l-dihydro-3-[3-(pyridin-2-ylamino)-l-propyloxy]-5H-5 dibenzo[a,d]cycloheptene-10-acetate <br><br> To a stirred solution of ethyl (S)-10,1 l-dihydro-3-[3-(l-oxopyndin-2-ylammo)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate (37 66 g, 84 mmol) in isopropanol (700 mL) were added 10% palladium on activated carbon (18 g, 16 9 mmol, carefully pre-wetted in isopropanol under argon) and cyclohexene (85 mL, 839 mmol) The reaction was 0 then heated to reflux under argon in an oil bath set at 90 °C After 6 h an additional amount of 10% palladium on activated carbon (18 g, 84 mmol, carefully pre-wetted in isopropanol under argon) and cyclohexene (85 mL, 839 mmol) were added After an additional 18 h the reaction was hot-filtered through celite®, and the filter pad was washed with 1.1 MeOH/CHCl3 (600 mL) The filtrate was concentrated under vacuum and the 5 residue was purified by flash chromatography on silica gel (95 5 CHC^/MeOH) to give the title compound (29 2 g, 81 %) as a pale yellow oil 'H NMR (400 MHz, DMSO-dg) 8 7.94 (dd, J = 5 4, 1.9 Hz, IH), 7 35-7.31 (m, IH), 7 18 (d, J = 7.2 Hz, IH), 7.14-7 06 (m, 3H), 6 97 (d, J = 8.3 Hz, IH), 6.83 (d, J = 2.6, IH), 6 68 (dd, J = 8 3, 2.6 Hz, IH), 6.54 (t, IH), 6 44 (m, 2H), 4 17 (d, J = 14 6 Hz, IH), 4 13-4 02 (m, 2H), 4 00 (t, 2H), 3 91 (d, J = 14 7 0 Hz, IH), 3.66 (m, IH), 3.35 (m, 2H), 3 19 (dd, J = 15 1,4 4 Hz, IH), 2 86 (dd, J = 15 1, 10 1 Hz, IH), 2 65 (dd, J = 15.8, 5.4 Hz, IH), 2.55 (dd, J = 15.8, 8 7 Hz, IH), 1.93 (m, 2H), 1 18 (t, 3H); MS (ES) m/e 431 4 (M + H)+ <br><br> c) (S)-10,ll-Dihydro-3-(3-(pyndin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetic acid <br><br> To a stirred solution of ethyl (S)-10,ll-dihydro-3-[3-(pyndin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate (29 20 g, 68 mmol) in dioxane (350 mL) was added aqueous 1 0 N NaOH (110 mL, 110 mmol) The cloudy reaction was stirred at 50 °C in an oil bath for 24 h, then the resulting homogeneous solution was neutralized with aqueous 1 0 N HCl (110 mL, 110 mmol). The solution was concentrated to near dryness by rotary evaporation to precipitate out the product. The supernatant was decanted off and the remaining gummy solid was dried under vacuum and redissolved in 1 1 methanol/CHCl3 The clear solution was then reconcentrated by rotary evaporation and thoroughly dried under vacuum The remaining solid was triturated with a small volume of water, filtered and dried under vacuum to give the title compound (26 85 g, 94%) as an off-white powder HPLC (Hamilton PRP-1®, 35% CH3CN/H2O containing 0 1% TFA) k' = 2.88, !H NMR (400 MHz, DMSO-d6) 8 7 94 (dd, J = 4.7, 1 6 Hz, IH), 7.38 (m, IH), 7 18 <br><br> -60- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> (d, J = 7 3 Hz, IH), 7 14 (d, J = 3.9 Hz, 2H), 7.08 (m, IH), 6 97 (d, J = 8 4 Hz, IH), 6 83 (d, 3 = 8 6 Hz, IH), 6 78 (br s, IH), 6 68 (dd, J = 8.3,2 6 Hz, IH), 6 50 (d, J = 8 3 Hz, IH), 6 47 (dd, IH), 4 20 (d, J = 14.6 Hz, IH), 4.00 (t, 2 H), 3 88 (d, J = 14 6 Hz, IH), 3 67 (m, IH), 3.37 (m, 1 H), 3.20 (dd, J = 15 2,4 4 Hz, IH), 2 83 (dd, J = 15 2,10 1 Hz, IH), 2 60 5 (dd, J = 15 9, 5.3 Hz, IH), 2 50 (dd, IH), 1 95 (m, 2 H), MS (ES) m/e 403 3 (M + H)+ <br><br> Anal Calcd for C25H26N2O3 H20 C, 71.41, H, 6 71, N, 6 66 Found* C, 71 21, H, 6.53, N, 6 54 <br><br> Example 5 <br><br> 10 <br><br> Preparation of (±)-10.11 -dihvdro-3-f2-(6-aminopvndin-2-vl')-1 -ethoxvl-5H-dibenzola.dlcvcloheptene-10-acetic acid a) Ethyl (±)-10,l l-dihydro-3-[2-(6-aminopyndin-2-yl)-l-ethoxy]-5H-15 dibenzo[a,d]cycloheptene-10-acetate <br><br> A solution of 6-amino-2-pyridylethanol (0 23 g, 1 68 mmole) and diethyl azodicarboxylate (0.26 mL, 1.68 mmole) in anhydrous DMF (5 mL) was added dropwise to a solution of ethyl (±)-10,l l-dihydro-3-hydroxy-5H-dibenzo[a,d]cycloheptene-10-acetate and tnphenylphosphme (0 48 g, 1 82 mmole) m anhydrous DMF (5 mL) at RT After 1 hr, 20 the reaction was concentrated and the residue was purified by flash chromatography on silica gel (1 1 EtOAc/hexanes) to afford the title compound (0 030 g) MS (ES) m/e 417 (M + H)+. <br><br> b) (±)-10,11 -Dihydro-3-[2-(6-aminopyridin-2-yl)-1 -ethoxy]-5H-dibenzo[a,d]cycloheptene-25 10-acetic acid <br><br> A solution of ethyl (±)-10,l l-dihydro-3-[2-(6-aminopyndin-2-yl)-l-ethoxy]-5H-dibenzo[a,d]cycloheptene-l 0-acetate (0.030 g, 0.072 mmole) and 1.0 N NaOH (0 14 mL, 0 14 mmole) in MeOH (2 mL) was stirred at RT overnight, then was concentrated The residue was dissolved in H2O and the pH of the solution was adjusted to 7 with 1.0 N HCl. 30 Concentration and chromatography on a C-18 Bond Elute column (10:9:1 <br><br> CH3CN/H20/TFA) gave the title compound (0.013 g). MS (ES) m/e 389 (M + H)+. <br><br> -61 - <br><br> Printed from Mimosa <br><br> WO 99/15508 PCT/US98/19466 <br><br> Example 6 <br><br> Preparation of (Rl-10.1 l-dihvdro-3-f3-(pvndm-2-vlaminoVI-propvloxvl-5H-dibenzora.dlcvcloheptene-10-acetic acid <br><br> 5 <br><br> a) Ethyl (R)-10,l l-dihydro-3-[3-(l-oxopyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-l 0-acetate <br><br> A solution of 2-[(3-hydroxy-l-propyl)amino]pyridine-N-oxide (0.70 g, 4 mmole) and diethyl azodicarboxylate (0.65 mL, 4 mmole) in anhydrous DMF (20 mL) was added 10 dropwise over 10 min to a solution of ethyl (R)-10,l l-dihydro-3-hydroxy-5H- <br><br> dibenzo[a,d]cycloheptene-l 0-acetate (0 45 g, 2 mmole) and tnphenylphosphme (1 2 g, 4 mmole) in anhydrous DMF (8 mL) at RT under argon After 23 5 hr, the reaction was concentrated on the rotavap, and the residue was reconcentrated from xylenes to remove residual DMF Silica gel chromatography (1-4% CH3OH/CH2CI2) gave the title 15 compound (0 50 g, 74 %) as a yellow oil. MS (ES) m/e 447 (M + H)+ <br><br> b) Ethyl (R)-10,l l-dihydro-3-[3-(pyndin-2-ylammo)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate <br><br> A mixture of ethyl (R)-10,l l-dihydro-3-[3-(l-oxopyridin-2-ylamino)-l-20 propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate (0 5 g, 1 mmole), 10% Pd/C (0 25 g, 0.2 mmole), cyclohexene (2 mL, 20 mmole), and isopropanol (10 mL) was heated at reflux for 18 hr, then the catalyst was removed by filtration through celite® Silica gel chromatography (0 5-2% CH3OH/CH2CI2) gave the title compound (0 4 g, 83%) as a light yellow oil: MS (ES) m/e 431 (M + H)+ <br><br> 25 <br><br> c) (R)-10,11 -Dihydro-3-[3-(pyridin-2-ylamino)-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene- 10-acetic acid <br><br> A mixture of ethyl (R)-10,l l-dihydro-3-[3-(pyndin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-l 0-acetate (0 4 g, .93 mmole) and 1.0 N NaOH (1 1 mL, 1.1 30 mmole) in absolute EtOH (10 mL) was warmed in an oil bath set at 50°C After 18 hr, the reaction was concentrated on the rotavap and the residue was dissolved in H2O. The aqueous solution was adjusted to pH 4 with 3 N HCl, and the solid precipitate was collected and washed with H2O The material was dried in high vacuum at 40°C to afford the title compound (0 36 g, 96%) as a nearly colorless solid. [a]Q +50 8° (c = 0.12, CH3OH), MS 35 (ES) m/e 403 (M + H)+. Anal. Calcd for C25H26N2O3 0.5 H20 C, 72 97, H, 6.61; N, 6.80 Found C, 73 09; H, 6 38, N, 6 58 <br><br> -62- <br><br> Printed from Mimosa <br><br> WO 99/15508 PCT/US98/19466 <br><br> Example 7 <br><br> Preparation of (SV10.1 l-dihvdro-3-(2-t6-(methvlanimo'&gt;pvTidin-2-vn-l-ethoxvl-5H-dibenzo[a.dlcvcloheptene-10-acetic acid <br><br> 5 <br><br> a) Ethyl (S)-10,l l-dihydro-3-[2-[6-(methylamino)pyridin-2-yl]-l-ethoxy]-5H-dibenzo[a,d]cycloheptene-10-acetate <br><br> According to the procedure of Example 6 (a), except substituting 6-(methylamino)-2-pyndylethanol for the 2-[(3-hydroxy-l-propyl)amino]pyndine-N-oxide, and ethyl (S)-10 10,1 l-dihydro-3-hydroxy-5H-dibenzo[a,d]cycloheptene-10-acetate for ethyl (R)-10,11-dihydro-3-hydroxy-5H-dibenzo[a,d]cycloheptene-10-acetate, the title compound was obtained as colorless oil following silica gel chromatography (0 2-2% MeOH/CH2Cl2)-MS (ES) 431.2 (M + H)+ <br><br> 15 b) (S)-10,11 -Dihydro-3-[2-[6-(methylamino)pyndin-2-y 1]-1 -ethoxy ]-5H-dibenzo[a,d]cycloheptene-10-acetic acid <br><br> Ethyl (S)-10,ll-dihydro-3-[2-[6-(methylamino)pyridin-2-yl]-l-ethoxy]-5H-dibenzo[a,d]cycloheptene-l 0-acetate (80 mg, 0.18 mmol) was dissolved in THF (4 mL), and a solution of LiOH H20 (35 mg, 0.84 mmol) in H20 (4 mL)was added The solution 20 was stirred at RT for 72 hr, then was diluted with ether (10 mL). The supernatant was decanted and the solid was suspended in H20. Careful acidification to pH 4 with 3 N HCl gave the title compound as a white solid MS(ES) 403 (M + H)+. Anal Calcd for C25H26N2°3 • 0 75 h20' c- 72 18; H, 6.66; N, 6 73 Found' C, 72 44, H, 6.52; N, 6 71. <br><br> 25 Example 8 <br><br> Preparation of (+V10.1 l-dihvdro-3-f3-f3.4.5.6-tetrahvdropvnmidin-2-vlaminoVl-propvloxvl-5H-dibenzora.dlcvcloheptene-10-acetic acid <br><br> 30 a) Ethyl (±)-10,l l-dihydro-3-[3-(4-nitrobenzyloxycarbonyl)amino-l-propy loxy ]-5H-dibenzo[a,d]cycloheptene-10-acetate <br><br> According to the procedure of Example 6 (a), except substituting 3-(4-nitrobenzyloxycarbonylammo)-l-propanol for the 2-[(3-hydroxy-l-propyl)amino]pyridine-N-oxide, and ethyl (±)-10,ll-dihydro-3-hydroxy-5H-dibenzo[a,d]cycloheptene-10-acetate 35 for ethyl (R)-10,1 l-dihydro-3-hydroxy-5H-dibenzo[a,d]cycloheptene-10-acetate, the title compound was obtained as amber oil. MS (ES) 533 3 (M + H)+ <br><br> -63- <br><br> Printed from Mimosa <br><br> WO 99/15508 PCT/US98/19466 <br><br> b) Ethyl (±)-10,l l-dihydro-3-(3-amino-l-propyloxy)-5H-dibenzo[a,d]cycloheptene-10-acetate <br><br> A mixture of ethyl (±)-10,l l-dihydro-3-[3-(4-nitrobenzyloxycarbonylammo)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-l 0-acetate (1 6 g, 3 mmol), 10% palladium on 5 charcoal (0 8 g, 1 mmol), and ethanol (50 mL) was shaken under H2 (48 psi) for 3 hr, then the catalyst was removed by filtration through celite® The filtrate was concentrated to give the title compound (1.2 g, 100%) as a yellow oil* MS (ES) 348 2 (M + H)+ <br><br> c) Ethyl (±)-10,l l-dihydro-3-[3-(pynmidin-2-ylamino)-l-propyloxy]-5H-10 dibenzo[a,d]cycloheptene-10-acetate <br><br> A mixture of ethyl (±)-10,11-dihydro-3-(3-amino-l-propyloxy)-5H-dibenzo[a,d]cycloheptene-l 0-acetate (0 4 g, 1 mmole), sodium bicarbonate (0 5 g, 6 mmole), 2-bromopyrimidine (0 34 g, 2 mmole) and ethanol (10 mL) was heated at reflux under argon for 18 hr The solution was then decanted and concentrated The residue was 15 purified by chromatography on silica gel (0 2 - 2% MeOH/CH2Cl2) to give the title compound (0.17 g, 34%) as a pale yellow oil MS (ES) 432.3 (M + H)+. <br><br> d) Ethyl (±)-10,l l-dihydro-3-[3-(3,4,5,6-tetrahydropynmid-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate <br><br> 20 A mixture of ethyl (+)-10,11 -dihydro-3-[3-(pynmidin-2-ylamino)-1 -propyloxy]- <br><br> 5H-dibenzo[a,d]cycloheptene-l0-acetate (0.17g, 0.38 mmol), 10% palladium on charcoal (0.085g, 0.08 mmol), 4 M HCl in dioxane (0.1 mL, 0.4 mmol) and ethanol (5 mL) was shaken under H2 (48 psi) for 6 hr, then the catalyst was removed by filtration through cehte®. The filtrate was concentrated to give the title compound (0 19 g) as a yellow oil 25 MS (ES) 436 3 (M + H)+. <br><br> e) (±)-10,11 -Dihydro-3-[3-(3,4,5,6-tetrahydropynmid-2-ylamino)-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetic acid <br><br> A solution of ethyl (±)-10,l l-dihydro-3-[3-(3,4,5,6-tetrahydropyrinud-2-ylamino)-30 l-propyloxy]-5H-dibenzo[a,d]cycloheptene-l 0-acetate (0 17g, 0.36 mmol), lithium hydroxide monohydrate (0.042g, 1 mmol), THF (3 mL), and water (10 mL) was stirred at room temperature for 20 hr, then was concentrated The residue was dissolved m water, and the solution was brought to pH 4 with 3 N HCl The resulting solution was kept in the refrigerator overnight, then the supernatant was decanted away from the solid. The solid 35 was dried in vacuum to give the title compound (0 145 g, 91 %) as a tan solid- MS (ES) 408 3 (M + H)+ Anal Calcd for C24H29N3O3 1.3 HCl C, 63 37, H, 6.71, N, 9 23 Found- C, 63.67; H, 6 84; N, 9 46 <br><br> -64- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> Example 9 <br><br> Preparation of (±)-10.1 l-dihvdro-3-[3-fisoauinoline-l-vlamino')-l-propvloxvl-5H-5 dibenzofa.dlcvcloheptene-1 Q-acetic acid a) Ethyl (±)-10,l l-dihydro-3-[3-(l-oxoisoquinoline-l-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate <br><br> According to the procedure of Example 6 (a), except substituting l-[(3-hydroxy-l-10 propyl)ammo]-isoquinoline N-oxide for the 2-[(3-hydroxy-l-propyl)aminoJpyndine-N-oxide, and substituting ethyl (±)-10,1 l-dihydro-3-hydroxy-5H-dibenzo[a,d]cycloheptene-10-acetate for the ethyl (R)-10,l l-dihydro-3-hydroxy-5H-dibenzo[a,d]cycloheptene-10-acetate, the title compound was prepared as a pale yellow oil. MS (ES) m/e 497 2 (M + H)+ <br><br> 15 <br><br> b) Ethyl (±)-10,11 -dihydro-3-[3-(isoqumoline-1 -ylamino)-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate <br><br> According to the procedure of Example 6 (b), except substituting ethyl (±)-10,l 1-dihydro-3-[3-( 1 -oxoisoquinoline-1 -ylamino)-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene-20 10-acetate for the ethyl (R)-10,l l-dihydro-3-[3-(l-oxopyndin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-l 0-acetate, the title compound was prepared as a clear oil MS (ES) m/e 481 3(M + H)+. <br><br> c) (±)-10,11 -Dihydro-3-[3-(isoquinoline-1 -ylamino)-1 -propyloxy]-5H-25 dibenzo[a,d]cycloheptene-10-acetic acid <br><br> According to the procedure of Example 6 (c), except substituting ethyl (±)-10,l 1-dihydro-3-[3-(isoquinoline-l-ylammo)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate for the ethyl (R)-10,l l-dihydro-3-[3-(pyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate, the title compound was prepared as an amber solid. 30 MS(ES)m/e453 2(M + H)+. Anal CalcdforC29H28N203 1.3 TFA 0 25 H20 C, 62 71, H, 4 96, N, 4 63. Found. C, 62 45, H, 4 92; N, 4 41 <br><br> -65- <br><br> Pnnted from Mimosa <br><br> WO 99/15508 PCT/US98/19466 <br><br> Example 10 <br><br> Preparation of (±&gt;10.1 l-dihvdro-3-f3-f4-('ethvlthio)Dvridin-2-vlaminol-l-propvloxvl-5H-dibenzola.dlcvcloheptene-10-acetic acid <br><br> 5 <br><br> a) Ethyl (±)-10,l l-dihydro-3-[3-[4-(ethylthio)-l-oxopyridin-2-ylamino]-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate <br><br> A solution of ethyl (±)-10,l l-dihydro-3-[3-(4-mtro-l-oxopyndin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene- 10-acetate (300 mg, 0 61 mmol) and sodium 10 thioethylate (145 mg, 1.22 mmol) in DMF (5 mL) was warmed at 70°C for 3h The solvent was removed on the rotavap and the residue was purified by silica gel chromatography (2 -6% CH3OH/CH2CI2) to give the title compound (90 mg) as an orange oil MS (ES) m/e 507.3(M + H)+ <br><br> 15 b) Ethyl (±)-10,11 -dihydro-3-[3-[4-(ethylthio)pyndin-2-ylamino]-1 -propyloxy ]-5H-dibenzo [a,d]cycloheptene-10-acetate <br><br> A mixture of ethyl (±)-10,l l-dihydro-3-[3-[4-(ethylthio)-l-oxopyndin-2-ylamino]-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate (60 mg, 0.119 mmol), Fe powder (70 mg), and glacial acetic acid (2 mL) was heated at 100°C for 1.5 h The mixture was 20 cooled to RT and diluted with H2O and EtOAc, and the pH was adjusted to 7 - 8 with solid Na2C03- The layers were separated, and the aqueous layer was extracted with EtOAc. The combined organic layers were washed with H2O, dried (MgSO^, and concentrated to give the title compound (60 mg) as a yellow oil MS (ES) m/e 491.3(M + H)+ <br><br> 25 c) (±)-10,l l-Dihydro-3-[3-[4-(ethylthio)pyridin-2-ylamino]-l-propyloxy]-5H-dibenzo [a,d]cycloheptene-10-acetic acid <br><br> According to the procedure of Example 6 (c), except substituting ethyl (±)-10,l 1-dihydro-3-[3-[4-(ethylthio)pyndin-2-ylammo]-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate for the ethyl (R)-10,l l-dihydro-3-[3-(pyndin-2-ylamino)-l-propyloxy]-5H-30 dibenzo[a,d]cycloheptene-10-acetate, the title compound was prepared as a yellow. 'H NMR (400 MHz, DMSO-d6) 5 7.77 - 7 76 (d, 1 H), 7 17 - 7.15 (d, 1 H), 7.13-7 12 (d, 2 H), 7.08 - 7.07 (m, 1 H), 6 96 - 6 94(d, 1 H), 6 81 - 6.80 (s, 1 H), 6.68 - 6 67 (d, 1 H), 6.52 (s, 1 H), 6.35 - 6 33 (d, 2 H), 6.30 (s, 1 H), 4.20 - 4.16 (d, 1 H), 3.99 - 3 96 (t, 2 H), 3.89 -3 85 (d, 1 H), 3.65 - 3 63 (m, 1 H), 3.36 - 3 32 (m, 2 H), 3 22 - 3 15 (m, 1 H), 2.96 - 2 90 35 (m, 2 H); 2.85 - 2.78 (m, 1 H), 2 62 - 2 56(m, 2 H), 1 94 - 1 90(m, 2 H), 1 26 - 1.22 (t, 3 H), MS (ES) m/e463 4(M + H)+ <br><br> -66- <br><br> Printed from Mimosa <br><br> WO 99/15508 PCT/US98/19466 <br><br> Example 11 <br><br> Preparation of (±)-!0.11 -dihvdro-2-methvl-3-l3-fpvndin-2-vlaminoV 1 -propvloxvl-5H-dibenzofa.dlcvcloheptene-10-acetic acid <br><br> 5 <br><br> a) Ethyl (±)-10,l 1 -dihydro-2-methyl-3-[3-[N-(/ert-butoxycarbonyl)-N-( 1 -oxopyridin-2-yl)amino]-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate <br><br> NaH (60% dispersion in mineral oil, 0 14 g, 0 37 mmol) was added to a solution of ethyl (±)-10,l l-dihydro-3-hydroxy-2-methyl-5H-dibenzo[a,d]cycloheptene-10-acetate (100 10 mg, 0 32 mmol) in DMSO (2 mL) under argon, and the reaction was stirred at RT for 0 5 hr A solution of 2-[N-(3-methanesulfonyloxy-1 -propyl)-N-(fert-butoxycarbonyl)amino]pyndine-N-oxide (160 mg, 0 4 mmole) in DMSO (1 mL) was then added dropwise The reaction was stirred at RT under argon for 18 hr, then was quenched with water (20 mL) and extracted with EtOAc. Drying (MgS04), concentration, and silica 15 gel chromatography (1 % MeOH/CH2Cl2) gave the title compound (85 mg, 42%) as a colorless oil. MS (ES) m/e 561 3 (M + H)+. <br><br> b) Ethyl (±)-10,l l-dihydro-2-methyl-3-[3-(l-oxopyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate <br><br> 20 TFA (0 16 g, 1 4 mmol) was added dropwise to a solution of ethyl (+)-10,l 1- <br><br> dihydro-2-methyl-3-[3-[N-(rerf-butoxycarbonyl)-N-( 1 -oxopyridin-2-yl)amino]-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate (80 mg, 0 14 mmole) in dry CH2CI2 (3 mL) The reaction was stirred for 5 hr then was concentrated on the rotavap to afford the title compound (60 mg, 43%) as a colorless oil MS (ES) m/e 461 1 (M + H)+. <br><br> 25 <br><br> c) Ethyl (±)-10,l l-dihydro-2-methyl-3-[3-(pyndin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate <br><br> According to the procedure of Example 6 (b), except substituting ethyl (±)-10,l 1-dihydro-2-methyl-3-[3-( 1 -oxopyridin-2-ylamino)-l -propyloxy]-5H- <br><br> 30 dibenzo[a,d]cycloheptene- 10-acetate for the ethyl (R)-10,l l-dihydro-3-[3-(l-oxopyndin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate, the title compound was prepared as an off-white solid. MS (ES) m/e 417.3 (M + H)+ <br><br> d) (±)-10,11 -Dihydro-2-methyl-3-[3-(pyndin-2-ylamino)-1 -propyloxy]-5H-35 dibenzo[a,d]cycloheptene-10-acetic acid <br><br> According to the procedure of Example 6 (c), except substituting ethyl (±)-10,l 1-dihydro-2-methyl-3-[3-(pyndin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene- <br><br> -67- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> 10-acetate for the ethyl (R)-10,l l-dihydro-3-[3-(pyridin-2-ylamino)-l-propy loxy ]-5H-dibenzo[a,d]cycloheptene- 10-acetate, the title compound was obtained as an off white solid 1H NMR (400 MHz, CDC13) 5 7 75 (d, 1 H), 7 65 (t, 1 H), 7.15 (m, 3 H), 7 05 (m, 1 H), 6 83 (s, 1 H), 6 7 (d, 1 H), 6 65 (m, 1 H), 6 60 (s, 1 H), 4.25 (d, J = 15 1 Hz, 1 H), 4.05 5 (t, 2 H), 3 80 (m, 1 H), 3 75 (d, J = 15 1 Hz, 1 H), 3.50 (t, 2 H), 3 25 (dd, 1 H), 2 85 (dd, 1 H), 2 68 (dd, 1 H), 2.60 (dd, 1 H), 2 15 (t, 2 H), 2.10 (s, 3 H), MS (ES) m/e 417 3 (M + <br><br> H)+ <br><br> Example 12 <br><br> 10 <br><br> Preparation of (+V10.11-dihvdro-2-fdimethvlamino)methvl-7-fluoro-3-r3-(pvridin-2-vlamino')-1 -propvloxv1-5H-dibenzora.d1cvcloheptene-10-acetic acid a) Ethyl (±)-10,l l-dihydro-2-(dimethylamino)methyl-7-fluoro-3-[3-(l-oxopyndin-2-15 ylamino)-1 -propy loxy]-5H-dibenzo[a,d]cycloheptene-10-acetate <br><br> According to the procedure of Example 6 (a), except substituting ethyl (±)-10,ll-dihydro-2-(dimethylamino)methyl-7-fluoro-3-hydroxy-5H-dibenzo[a,d]cycloheptene-10-acetate for the ethyl (R)-10,11 -dihydro-3-hydroxy-5H-dibenzo[a,d]cycloheptene-10-acetate ethyl, the title compound was obtained following silica gel chromatography (gradient: 1 1 20 EtOAc hexanes, then EtOAc, then 20% MeOH/CH2Cl2, then 30% MeOH/C^Cy MS (ES) m/e 522 3 (M + H)+ <br><br> b) Ethyl (±)-10,l l-dihydro-2-(dimethylamino)methyl-7-fluoro-3-[3-(pyndin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate <br><br> 25 According to the procedure of Example 6 (b), except substituting ethyl (±)-10,11 - <br><br> dihydro-2-(dimethylamino)methyl-7-fluoro-3-[3-(l-oxopyndin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate for the ethyl (R)-10,11 -dihydro-3-[3-( 1 -oxopyndin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate, the title compound was obtained following silica gel chromatography (10% MeOH/CH2Cl2) MS 30 (ES) m/e 506 2 (M + H)+ <br><br> c) (±)-10,1 l-Dihydro-2-(dimethylamino)methyl-7-fluoro-3-[3-(pyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetic acid <br><br> Saponification was conducted according to the procedure of Example 6 (c), except 35 substituting ethyl (±)-10,l l-dihydro-2-(diniethylamino)methyl-7-fluoro-3-[3-(pyndin-2-ylamino)-l-propy loxy ]-5H-dibenzo[a,d]cycloheptene- 10-acetate for the ethyl (R)-10,l 1-dihydro-3-[3-(pyndm-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate <br><br> -68- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> The reaction was acidified with glacial HOAc, and the crude product was desalted by chromatography on XAD-2 resin to afford the title compound as a white solid MS (ES) m/e 478.3 (M + H)+. Anal. Calcd for C3oH36FN305 1 25 H20. C, 64 32, H, 6 92; N, 7 50 Found C, 63 87; H, 6 47, N, 7 96 <br><br> 5 <br><br> Example 13 <br><br> Preparation of CSI-lO.l l-dihvdro-3-[3-("4-methvlpvridin-2-vlaminoVl-propvloxvl-5H-dibenzora.dlcvcloheptene-10-acetic acid <br><br> 10 <br><br> a) Ethyl (S)-10,l l-dihydro-3-[3-(4-methyl-l-oxopyndin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate <br><br> A solution of 2-[(3-hydroxy-l-propyl)amino]-4-methylpyridine-N-oxide (1 72 g, 9 45 mmole) and diethyl azodicarboxylate (1 49 mL, 9 45 mmole) in anhydrous DMF (50 15 mL) was added dropwise over 10 min to a solution of ethyl (S)-10,l l-dihydro-3-hydroxy-5H-dibenzo[a,d]cycloheptene- 10-acetate (1.4 g, 4 72 mmole) and tnphenylphosphme (2 60 g, 9 92 mmole) in anhydrous DMF (50 mL) at RT under argon. After 19 hr, the reaction was concentrated on the rotavap, and the residue was reconcentrated from xylenes to remove residual DMF. Silica gel chromatography (gradient- 30% EtOAc/hexanes (0.5 L), 20 then EtOAc (1 L), then 5% MeOH/CHCl3) gave the title compound (1 31 g, 60 %) as a yellow oil MS (ES) m/e 461 3 (M + H)+. <br><br> b) Ethyl (S)-10,l l-dihydro-3-[3-(4-methylpyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate <br><br> 25 A mixture of ethyl (S)-10,l l-dihydro-3-[3-(4-methyl-l-oxopyndin-2-ylamino)-l- <br><br> propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate (0 86 g, 1 87 mmole), 10% Pd/C (0 86 g, 0.81 mmole), cyclohexene (1 89 mL, 18 7 mmole), and isopropanol (20 mL) was heated at reflux under argon for 19 hr, then the catalyst was removed by filtration through cehte® Silica gel chromatography (19 10 MeOH/C^C^/EtOAc) gave the title compound (0 65 30 g, 78%) as a clear oil MS (ES) m/e 445.2 (M + H)+. <br><br> c) (S)-10,11 -Dihydro-3-[3-(4-methylpyridin-2-ylamino)-1-propy loxy ]-5H-dibenzo[a,d]cycloheptene-10-acetic acid <br><br> A mixture of ethyl (S)-10,ll-dihydro-3-[3-(4-methylpyridin-2-ylamino)-l-35 propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate (2 08 g, 4 69 mmole) and 1 0 N NaOH (7.0 mL, 7 0 mmole) in absolute EtOH (45 mL) was warmed in an oil bath set at 45°C After 18 hr, the reaction was concentrated on the rotavap and the pH was adjusted to 7 with <br><br> -69- <br><br> Prmted from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> 1 0 N HCl. The solid precipitate was collected and washed with H20 Drying overnight afforded the title compound (1 61 g, 82%) as a nearly colorless solid- MS (ES) m/e 417 4 (M + H)+ Anal. Calcd for C26H28N2O3 1 0 H20: C, 71.87; H, 6 96, N, 6 45 Found. C, 71 63, H, 6 96, N, 6.30 <br><br> 5 <br><br> Example 14 <br><br> Preparation of fSVlO.l l-dihvdro-3-13-r4-&lt;,2-proDvloxv')pvridin-2-vlamino1-l-propvloxv1-5H-dibenzora.d1cvcloheptene-10-acetic acid <br><br> 10 <br><br> a) Isopropyl (S)-10,l l-dihydro-3-[3-[4-(2-propyloxy)-l-oxopyridin-2-ylamino]-l-propy loxy ]-5H-dibenzo[a,d]cycloheptene-10-acetate <br><br> A mixture of ethyl (S)-10,l l-dihydro-3-[3-(4-nitro-l-oxopyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate (0 2 g, 0 4 mmole) and sodium 15 isopropoxide (0 067 g, 0 8 mmole) in isopropanol (5 mL) was heated at 80°C for 3 5 hr, <br><br> then more sodium isopropoxide (0.05 g, 0.6 mmole) was added, and the reaction was stirred at RT overnight Concentration and silica gel chromatography (gradient 5% - 15% MeOH/CH2Cl2) gave the title compound (0 106 g, 52%) as a light brown oil. MS (ES) 519 3 (M + H)+. <br><br> 20 <br><br> b) Isopropyl (S)-10,l l-dihydro-3-[3-[4-(2-propyloxy)pyridin-2-ylamino]-l-propyloxy]-5H-dibenzo[a,d]cy cloheptene-10-acetate <br><br> According to the procedure of Example 13 (b), except substituting isopropyl (S)-10,1 l-dihydro-3-[3-[4-(2-propyloxy)-1 -oxopyridin-2-ylamino]-1 -propy loxy]-5H-25 dibenzo[a,d]cycloheptene-10-acetate for the ethyl (S)-10,11 -dihydro-3-[3-(4-methyl-1 -oxopyndin-2-ylammo)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate, the title compound was obtained as slightly yellow oil following silica gel chromatography (5% MeOH/CH2Cl2) MS (ES) 503 4 (M + H)+. <br><br> 30 c) (S)-10,ll-Dihydro-3-[3-[4-(2-propyloxy)pyndin-2-ylammo]-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetic acid <br><br> According to the procedure of Example 13 (c), except substituting isopropyl (S)-10,1 l-dihydro-3-[3-[4-(2-propyloxy)pyndin-2-ylamino]-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate for the ethyl (S)-10,l l-dihydro-3-[3-(4-35 methylpyndm-2-ylamino)-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate, the title compound was obtained as white powder. MS (ES) 461.3 (M + H)+ Anal Calcd for C28H32N2°4 0 96 HCl C, 67 86, H, 6 70, N, 5 65 Found C, 68.26, H, 6 86, N, 5 25 <br><br> -70- <br><br> Printed from Mimosa <br><br> WO 99/15508 PCT/US98/19466 <br><br> Example 15 <br><br> Preparation of (S)-lO.l l-dihvdro-3-r3-(4-chloropvndin-2-vlamino)-l-propyloxv1-5H-5 dibenzora.dlcvcloheptene-10-acetic acid a) Ethyl (S)-10,l l-dihydro-3-[3-(4-chloro-l-oxopyndin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate <br><br> A solution of ethyl (S)-10,1 l-dihydro-3-[3-(4-nitro-l-oxopyridin-2-ylamino)-l-10 propyloxy]-5H-dibenzo[a,d]cycloheptene- 10-acetate (0 47 g, 0 96 mmole) in acetyl chloride (7 mL, 98 mmole) was heated at reflux for 1 hr The reaction mixture was poured onto ice (50 g), and the pH was adjusted to 8.0 using saturated NaHC03 (caution bubbles violently1) The mixture was extracted with CH2CI2 (2 x 100 mL), and the combined organic layers were washed sequentially with H2O (50 mL) and brine (50 mL) Drying 15 (MgSC&gt;4) and concentration gave the title compound MS (ES) 481.2 (M + H)+ <br><br> b) Ethyl (S)-10,1 l-dihydro-3-[3-(4-chloropyridin-2-ylamino)-l-propyloxy]-5H-dibenzo [a,d]cycloheptene-10-acetate <br><br> A mixture of ethyl (S)-10,l l-dihydro-3-[3-(4-chloro-l-oxopyndin-2-ylamino)-l-20 propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate (0 13 g, 0 27 mmole) and 2 0 M PCI3 in CH2CI2 (8 mL, 16 mmole) was heated at reflux for 22 hr The reaction mixture was cooled and poured onto ice (200 g), and the pH was adjusted to 12 using 40% NaOH CH2CI2 (2 x 100 mL) extraction, drying (MgS04), concentration, and silica gel chromatography (4% MeOH/C^Cl^ gave the title compound (93 mg, 74%) as light 25 yellow oil. MS (ES) 465 3 (M + H)+ <br><br> c) (S)-10,11 -Dihydro-3-[3-(4-chloropyridin-2-ylamino)-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene- 10-acetic acid <br><br> According to the procedure of Example 13 (c), except substituting ethyl (S)-10,11-30 dihydro-3-[3-(4-chloropyndin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate for the ethyl (S)-10,1 l-dihydro-3-[3-(4-methylpyridin-2-ylammo)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate, the title compound was obtained as off-white powder MS (ES) 437 2 (M + H)+. Anal. Calcd for C25H25N203 1 0 HCl C, 63.43; H, 5 54, N, 5 92. Found C, 63 11, H, 5 82, N, 5.62 <br><br> 35 <br><br> -71 - <br><br> Printed from Mimosa <br><br> WO 99/15508 PCT/US98/19466 <br><br> Example 16 <br><br> Preparation of (SV10.1 l-dihvdro-3-r3-f4-("dimethvlamino)pvndin-2-vlaminol-l-propvloxvl-5H-dibenzo[a.dlcvcloheptene-10-acetic acid <br><br> 5 <br><br> a) Ethyl (S)-1Q,1 l-dihydro-3-[3-(4-chloro-l-oxopyndm-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate <br><br> A solution of ethyl (±)-10,l l-dihydro-3-[3-(4-nitro-l-oxopyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate (0.47 g, 0.96 mmole) in acetyl 10 chloride (7 mL, 98 mmole) was heated at reflux for 1 hr The reaction mixture was poured onto ice (50 g), and the pH was adjusted to 8 0 using saturated NaHCC&gt;3 (caution- bubbles violently') The mixture was extracted with CH2CI2 (2 x 100 mL), and the combined organic layers were washed sequentially with H2O (50 mL) and brine (50 mL) Drying (MgSCty) and concentration gave crude title compound which was carried forward without 15 further purification MS (ES) 481.3 (M + H)+ <br><br> b) Ethyl (S)-10,l l-dihydro-3-[3-[4-(dimethylamino)-l-oxopyridin-2-ylammo]-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate <br><br> A mixture of ethyl (S)-10,l l-dihydro-3-[3-(4-chloro-l-oxopyridin-2-ylamino)-l-20 propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate (0 96 mmole) and 2 0 M <br><br> dimethylamine in MeOH (3 mL, 6 mmole) was refluxed for 16 hr. Concentration and silica gel chromatography (7% MeOH/CH2Cl2) gave the title compound (0 049 g, 10 %) as a light brown powder MS (ES) 490.3 (M + H)+ Unchanged ethyl (S)-10,l l-dihydro-3-[3-(4-chloro-1 -oxopyridin-2-yiamino)-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate 25 was also recovered from the chromatographic purification c) Ethyl (S)-10,l l-dihydro-3-[3-[4-(dimethylamino)pyridin-2-ylamino]-l -propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate <br><br> According to the procedure of Example 13 (b), except substituting ethyl (S)-10,11 -30 dihydro-3-[3-[4-(dimethylamino)-l-oxopyridin-2-ylamino]-l-propyloxy]-5H- <br><br> dibenzo[a,d]cycloheptene-10-acetate for the ethyl (S)-10,l l-dihydro-3-[3-(4-methyl-l-oxopyridin-2-ylammo)-1 -propyloxy ]-5H-dibenzo[a,d]cycloheptene-10-acetate, the title compound was obtained as white powder following silica gel chromatography (8% MeOH/CH2Cl2)' MS (ES) 474.3 (M + H)+ <br><br> 35 <br><br> -72- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> d) (S)-10,11 -Dihydro-3-[3-[4-(dimethylamino)pyndm-2-ylamino]-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetic acid <br><br> According to the procedure of Example 13 (c), except substituting ethyl (S)-10,11-dihydro-3-[3-[4-(dimethylamino)pyridin-2-ylamino]-l-propyloxy]-5H-5 dibenzo[a,d]cycloheptene-10-acetate for the ethyl (S)-10,1 l-dihydro-3-[3-(4- <br><br> methylpyndin-2-ylamino)-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate, the title compound was obtained as white powder' MS (ES) 446 2 (M + H)+ Anal Calcd for c27h3 1n3°3 ' 0 5 h2° 1 0 HCI C, 66 04, H, 6 77, N, 8.56 Found: C, 65 96, H, 6.60, N, 8.26 <br><br> 10 <br><br> Example 17 <br><br> Preparation of (S)-IO.I1 -dihvdro-3-l3-C4-ethoxvpvridin-2-vlamino')-1 -propvloxv1-5H-dibenzofa.dlcvcloheptene-10-acetic acid <br><br> 15 <br><br> a) Ethyl (S)-10,11 -dihydro-3-[3-(4-ethoxy-1 -oxopyndin-2-ylamino)-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate <br><br> According to the procedure of Example 2 (a), except substituting ethyl (S)-10,l 1-dihydro-3-[3-(4-nitro-l-oxopyridin-2-ylamino)-l-propyloxy]-5H- <br><br> 20 dibenzo[a,d]cycloheptene- 10-acetate (496.9 mg, 1 01 mmol) for the ethyl (±)-10,l 1-dihydro-3-[3-(4-nitro-1 -oxopyndin-2-ylamino)-1 -propyloxy]-5H-dibenzo[a,d]cyc!oheptene- 10-acetate, and using 0 53 M NaOEt (4.0 mL, 2 12 mmol) and absolute ethanol (10 mL) in the displacement reaction, the title compound (456 2 mg, 92%) was prepared: MS (ES) m/e 491 (M + H)+. <br><br> 25 <br><br> b) Ethyl (S)-10,l 1-dihydro-3-[3-(4-ethoxypyridm-2-ylamino)-1-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate <br><br> According to the procedure of Example 2 (b), except substituting ethyl (S)-10,l 1-dihydro-3- [3-(4-ethoxy-1 -ox opyridm-2-y lamino)-1 -propy loxy ]-5H- <br><br> 30 dibenzo[a,d]cycloheptene-10-acetate (456 2 mg, 0 93 mmole) for the ethyl (+)-10,l 1-dihydro-3-[3-(4-ethoxy-l-oxopyndin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate, the title compound (475.2 mg, quantitative) was prepared MS (ES) m/e 475 (M + H)+ <br><br> -73- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> c) (S)-10,11 -Dihydro-3-[3-(4-ethoxypyridin-2-ylamino)-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetic acid <br><br> ION NaOH (2 0 mL, 2 0 mmole) was added to a solution of ethyl (S)-10,l 1-dihydro-3-[3-(4-ethoxypyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-5 acetate (475 2 mg, 1 0 mmol) in absolute ethanol (10 mL), and the solution was warmed at 50°C in an oil bath After 20 hr, the reaction was concentrated and the aqueous residue was cooled to 0°C in an ice bath. 1 0 N aqueous HCl (2 0 mL, 2 0 mmole) was slowly added with stirring An opaque solid residue precipitated and was collected on a sintered glass funnel. Drying in a vacuum desiccator overnight gave the title compound (452.6 mg, 83%) 10 MS (ES) m/e 447 (M + H)+ Anal Calcd for C27H30N2O4 0.20 H20 1.75 HCl. C, 63.10, H, 6 30; N, 5 45 Found C, 63.10; H, 5 98, N, 5.38 <br><br> Example 18 <br><br> 15 Preparation of (±V10.1 l-dihvdro-7-fluoro-3-f3-(pvndin-2-vlamino'l-l-propvloxvl-5H-dibenzofa.dlcvcloheptene-10-acetic acid a) Ethyl (±)-10,l l-dihydro-7-fluoro-3-[3-(l-oxopyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate <br><br> 20 According to the procedure of Example 6 (a), except substituting ethyl (±)-10,11 - <br><br> dihydro-7-fluoro-3-hydroxy-5H-dibenzo[a,d]cycloheptene-10-acetate for the ethyl (R)-10,1 l-dihydro-3-hydroxy-5H-dibenzo[a,d]cycloheptene-10-acetate, the title compound was obtained as a colorless oil following silica gel chromatography (gradient 1 1 EtOAc/hexanes, then EtOAc, then 4% MeOH/CH2Cl2): MS (ES) m/e 465 3 (M + H)+. <br><br> 25 <br><br> b) Ethyl (±)-10,11 -dihydro-7-fluoro-3-[3-(pyridin-2-ylamino)-1 -propyloxy]-5H-dibenzo[ a,d]cycloheptene-l 0-acetate <br><br> According to the procedure of Example 6 (b), except substituting ethyl (±)-10,l 1-dihydro-7-fluoro-3-[3-(l-oxopyndin-2-ylamino)-l-propyloxy]-5H-30 dibenzo[a,d]cycloheptene-10-acetate for the ethyl (R)-10,11 -dihydro-3-[3-( 1 -oxopyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate, the title compound was obtained: MS (ES) m/e 449.2 (M + H)+ <br><br> c) (±)-10,11 -Dihydro-7-fluoro-3-[3-(pyndin-2-ylammo)-1 -propyloxy]-5H-35 dibenzo[a,d]cycloheptene-10-acetic acid <br><br> According to the procedure of Example 6 (c), except substituting ethyl (+)-10,11-dihydro-7-fluoro-3-[3-(pyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10- <br><br> -74- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> acetate for the ethyl (R)-10,l l-dihydro-3-[3-(pyridin-2-ylamino)-l-propy loxy ]-5H-dibenzo[a,d]cycloheptene-10-acetate, the title compound was obtained MS (ES) m/e 421 1 (M + H)+ Anal Calcd for C25H25FN2O3 0 5 H20 C, 69.99, H, 6 10, N, 6 52 Found C, 69.86, H, 5 90, N, 6.35. <br><br> 5 <br><br> Example 19 <br><br> Preparation of (±&gt;-10.1 l-dihvdro-6-methvl-3-r3-(pvndin-2-vlamino'&gt;-l-propvloxv1-5H-dibenzora.dlcvcloheptene-10-acetic acid <br><br> 10 <br><br> a) Ethyl (±)-10,l l-dihydro-6-methyl-3-[3-(l-oxopyndin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate <br><br> According to the procedure of Example 6 (a), except substituting ethyl (±)-10,l 1-dihydro-3-hydroxy-6-methyl-5H-dibenzo[a,d]cycloheptene-10-acetate for the ethyl (R)-15 10,11 -dihydro-3-hydroxy-5H-dibenzo[a,d]cycloheptene-10-acetate, the title compound was obtained as a colorless oil following silica gel chromatography (gradient: 1 1 EtOAc/hexanes, then EtOAc, then 4% MeOH/CH2Cl2). MS (ES) m/e 461 3 (M + H)+ <br><br> b) Ethyl (±)-10,1 l-dihydro-6-methyl-3-[3-(pyridin-2-ylamino)-l-propyloxy]-5H-20 dibenzo[a,d]cycloheptene-10-acetate <br><br> According to the procedure of Example 6 (b), except substituting ethyl (±)-10,l 1-dihy dro-6-methyl-3-[3-( 1 -oxopyridin-2-ylamino)-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate for the ethyl (R)-10.1 l-dihydro-3-[3-(l-oxopyndin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene- 10-acetate, the title compound was 25 obtained following silica gel chromatography (1% MeOH/CH2Cl2): MS (ES) m/e 445.3 (M + H)+ <br><br> c) (±)-10,1 l-Dihydro-6-methyl-3-[3-(pyndin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetic acid <br><br> 30 According to the procedure of Example 6 (c), except substituting ethyl (±)-10,l 1- <br><br> dihydro-6-methyl-3-[3-(pyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate for the ethyl (R)-10,l l-dihydro-3-[3-(pyndin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate, the title compound was obtained as a white solid MS (ES) m/e 417 3 (M + H)+ Anal. Calcd for C26H28N2O3 1.25 H20 C, 71.13, H, 35 7 02; N, 6 38 Found: C, 71.33, H, 6 67, N, 6 01. <br><br> -75- <br><br> Printed from Mimosa <br><br> WO 99/15508 PCT/US98/19466 <br><br> Example 20 <br><br> Preparation of CSV 10.1 l-dihvdro-3-[3-(4-aminoPvridin-2-vlaminoVl-propvloxvl-5H-5 dibenzofa.dlcvcloheptene-10-acetic acid a) Ethyl (S)-10,11 -dihydro-3-[3-(4-nitro-1 -oxopyridin-2-ylamino)-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate <br><br> A solution of dnsopropyl azodicarboxylate (1 7 mL, 8 mmole) in THF (10 mL) was 10 added dropwise to a solution of ethyl (S)-10,l l-dihydro-3-hydroxy-5H- <br><br> dibenzo[a,d]cycloheptene-10-acetate (426 5 mg, 1.5 mmole), 2-[(3-hydroxy-l-propyl)amino]-4-nitropyndine-N-oxide (1 7 g, 8 mmole), and tnphenylphosphme (2 5 g, 8 mmole) in anhydrous DMF (20 mL) at 0 °C under argon The yellow solution was kept at 0 °C for 10 mm, then was warmed to RT. After 23 hr, the reaction was concentrated Silica 15 gel chromatography (gradient. 30% - 100% EtOAc/hexanes) gave the title compound (2 7 g, 81%) as an orange foam: MS (ES) m/e 491 8 (M + H)+. <br><br> b) Ethyl (S)-10,11 -dihydro-3-[3-(4-aminopyridin-2-ylamino)-1 -propyloxy]-5H-dibenzo[a,d]cy cloheptene-10-acetate <br><br> 20 A mixture of ethyl (S)-10,l l-dihydro-3-[3-(4-nitro-l-oxopyndin-2-ylamino)-l- <br><br> propyloxyJ-5H-dibenzo[a,d]cycloheptene-10-acetate (2 7 g, 6 mmole), cyclohexene (6 mL, 60 mmole), 10% Pd/C (1 2 g, 1.10 mmole), and isopropanol (30 mL) was heated at reflux under argon for 20 5 hr, then was hot-filtered through cehte® The filter pad was washed with hot EtOAc, and the combined filtrates were concentrated The residue was 25 chromatographed on silica gel (5% MeOH/CHCl3) to afford the title compound (2.4 g, 98%) as a colorless foam: MS (ES) m/e 445.9 (M + H)+. <br><br> c) (S)-10,11 -Dihydro-3-[3-(4-aminopyndin-2-ylamino)-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetic acid <br><br> 30 A mixture of ethyl (S)-10,l l-dihydro-3-[3-(4-aminopyridin-2-ylamino)-l- <br><br> propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetate (2.4 g, 5 mmole), LiOH • H2O (0 3 g, 7 mmole), THF (30 mL), and H2O (10 mL) was stirred at RT for 48 hr, then was concentrated The residue was diluted with H20 and extracted with Et20 The Et20 layers were discarded. The aqueous layer was stirred with gentle warming under vacuum to 35 remove residual organic solvents, then was filtered. The resulting aqueous solution was stirred at RT while the pH was slowly and carefully adjusted to 5.5 - 6 0 with 1.0 N HCl The mixture was stirred for 0 5 hr, then the solid was collected by suction filtration and <br><br> -76- <br><br> Printed from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> washed with plenty of H2O Drying in high vacuum at 60 °C gave the title compound (1 0 g, 42%) as a glassy solid MS (ES) m/e 417.7 (M + H)+ Anal Calcd for C25H27N3O3 1 4 HCl (468 554) C, 64 08; H, 6 11; N, 8 97. Found C, 64.16, H, 6 20, N, 8 71 <br><br> 5 Example 21 <br><br> Preparation of (±1-10.1 l-dihvdro-3-r3-('4-methvlpvridin-2-vlaminoV I-pronvloxvldibenzorb.floxepine-10-acetic acid <br><br> 10 a) Ethyl (±)-10,l 1-dihydro-3-[3-(4-methyl-l-oxopyridin-2-ylamino)-l-propyloxy]dibenzo[b,f]oxepme-10-acetate <br><br> A mixture of ethyl (±)-10,ll-dihydro-3-hydroxydibenzo[b,f]oxepine-10-acetate (257 mg, 0.86 mmole), 2-[(3-bromo-l-propyl)amino]-4-methylpyridine-N-oxide hydrobromide (308 mg, 0.94 mmole), NaOH pellets (110 mg, 2 75 mmole), and CH3CN (4 15 mL) was stirred at RT under argon overnight The mixture was filtered and the solids were washed with CH3CN. The filtrate was concentrated, and the residue was flash chromatographed on silica gel (1 - 2.5% CH3OH/CH2CI2) to afford the title compound (190 mg, 48%) as a white foam MS (ES) m/e 462 6 (M + H)+ <br><br> 20 b) Ethyl (±)-10,11 -dihydro-3-[3-(4-methylpyridin-2-ylamino)-1 -propyloxy]dibenzo[b,f]oxepine-10-acetate <br><br> A mixture of ethyl (±)-10,ll-dihydro-3-[3-(4-methyl-l-oxopyridin-2-ylamino)-l-propyloxy]dibenzo[b,f]oxepine-10-acetate (183 mg, 0 4 mmole), 10% Pd/C (85 mg, 0 08 mmole), cyclohexene (810 mg, 8 mmole), and isopropanol (4 mL) was heated at reflux 25 overnight. The catalyst was removed by filtration through cehte®, and the filter cake was washed with ether. The filtrate was concentrated to afford the title compound (122 mg, 68%) as a clear oil MS (ES) m/e 446 9 (M + H)+. <br><br> c) (±)-10,11 -Dihydro-3-[3-(4-methylpyndin-2-ylamino)-1 -propyloxy]dibenzo[b,f]oxepine-30 10-acetic acid <br><br> A mixture of ethyl (±)-10,l l-dihydro-3-[3-(4-methylpyridin-2-ylamino)-l-propyloxy]dibenzo[b,f]oxepme-10-acetate (119 mg, 0 27 mmole) and 0 991 N NaOH (0 545 mL, 0 54 mmole) in absolute EtOH (2 mL) was warmed in an oil bath set at 45 °C After 20 hr, the reaction was concentrated on the rotavap, and the residue was dissolved in 35 H2O (1 5 mL) The solution was filtered to remove insoluble material, and the filtrate was carefully neutralized by dropwise addition of 1 0 N HCl (0.54 mL, 0 54 mmole) The precipitate was collected and dried m high vacuum to afford the title compound (68 mg, <br><br> -77- <br><br> Prxnted from Mimosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> 58%) as a white solid MS (ES) m/e 418 9 (M + H)+. Anal Calcd for C25H26N2O4 0 45 HCl C, 69.05, H, 6 13, N, 6 44 Found C, 69 25, H, 6 27, N, 6 16 <br><br> Example 22 <br><br> 5 <br><br> Preparation of (±"&gt;-10.1 l-dihvdro-3-r2-r6-(niethvlamino')pvridin-2-vP-l-ethoxvldibenzorb.floxepine-10-acetic acid a) Ethyl (±)-10,l l-dihydro-3-[2-[6-[N-tert-butoxycarbonyl)-N-methylamino]pyridin-2-yl]-10 1 -ethoxy]dibenzo[b,f]oxepine-10-acetate <br><br> A solution of 6-[N-(tert-butoxycarbonyl)-N-methylamino]-2-pyndylethanol (397 mg, 1 58 mmole) and dnsopropyl azodicarboxylate (0.31 mL, 1.58 mmole) in anhydrous CH2CI2 (8 mL) was added dropwise over 10 min to a solution of ethyl (±)-10,l 1-dihydro-3-hydroxydibenzo[b,f]oxepine-10-acetate (186 mg, 0.63 mmole) and tnphenylphosphme 15 (413 mg, 1 58 mmole) in anhydrous CH2CI2 (3 2 mL) at RT under argon. After 22 hr, the reaction was concentrated on the rotavap, and the residue was flash chromatographed on silica gel (2 -13% EtOAc/hexanes) to give the title compound (146 mg, 44%) as a clear oil MS (ES) m/e 533 0 (M+ H)+. <br><br> 20 b) Ethyl (±)-10,l l-dihydro-3-[2-[6-(methylamino)pyridin-2-yl]-l-ethoxy]dibenzo[b,f]oxepine-10-acetic acid <br><br> 4 N HCl in dioxane (1 3 mL, 5.2 mmole) was added dropwise to a solution of ethyl (±)-10,11 -dihydro-3-[2-[6-[N-tert-butoxycarbonyl)-N-methylamino]pyndin-2-yl]-1 -ethoxy]dibenzo[b,f]oxepine-10-acetate (140 mg, 0.26 mmole) in CH2CI2 (1.3 mL) After 25 12 hr, the mixture was concentrated, and the residue was triturated with ether to afford the title compound as a white solid MS (ES) m/e 432.9 (M + H)+. <br><br> c) (±)-10,11 -Dihydro-3-[2-[6-(methylamino)pyridin-2-yl]-1 -ethoxy ]dibenzo[b,f]oxepine-10-acetic acid <br><br> 30 A mixture of ethyl (±)-10,l l-dihydro-3-[2-[6-(methylamino)pyndin-2-yl]-l- <br><br> ethoxy]dibenzo[b,f]oxepine-10-acetic acid (0.26 mmole) and 0.991 N NaOH (0 525 mL, 0 52 mmole) in absolute EtOH (2 mL) was warmed in an oil bath set at 50 °C. After 20 hr, the reaction was concentrated on the rotavap and the residue was dissolved in H2O (1 5 mL) The solution was filtered to remove insoluble material, and the filtrate was carefully 35 neutralized by dropwise addition of 1 0 N HCl. The precipitate was collected and dried m high vacuum to afford the title compound (72 mg, 30% for 2 steps) as an off-white solid <br><br> -78- <br><br> Prmted from Mxmosa <br><br> WO 99/15508 <br><br> PCT/US98/19466 <br><br> MS (ES) m/e 405 0 (M + H)+ Anal Calcd for C24H24N2O4 1 25 HCl 0 25 H20 C, 63.42, H, 5 71, N, 6 16. Found. C, 63 35, H, 5 9, N, 6 16 <br><br> Example 23 <br><br> 5 <br><br> Preparation of (51-10.1 l-dihvdro-3-r3-(2-aminopvridin-4-vl')-l-propvloxv1-5ff-dibenzola.&lt;ikvcloheptene-10-acetic acid a) 3-(2-Ammopyridin-4-yl)propan-1 -ol <br><br> 10 A suspension of 3-(2-aminopyndin-4-yl)propanoic acid hydrochloride (0 73 g, 3 60 <br><br> mmol, prepared according to W094/14776 in THF (10 mL) was added over 45 mm to lithium aluminum hydride (12 mL, 12 mmol, 1M in THF) at 0 °C The ice bath was removed and the reaction was allowed to stir at RT for 4.5 h The reaction was cooled to 0 °C, diluted with toluene (22 mL) and quenched by the sequential addition of H20 (0.86 mL) 15 and NaF (1.54 g) The resulting suspension was stirred at 0 °C for 45 min. The reaction mixture was filtered and the precipitate was washed with additional 10% MeOH in CHC1, The combined filtrates were concentrated under reduced pressure. Flash chromatography (10% MeOH/CHCl,, silica gel) gave 0 25 g of the desired material as a clear oil: MS(ES+) m/z 152.7 [M+H]' <br><br> 20 <br><br> b) Ethyl (S)-10,l 1-dihydro-3-[3-(2-aminopyndm-4-yl)-1-propyloxy]-5//-dibenzo[a,&lt;f]cycloheptene-10-acetate <br><br> A solution of Example 1(a) (0 23 g, 1 51 mmol) and di-isopropylazadicarboxylate (0 29 mL, 1.50 mmol) in CH2C12 (7.5 mL) was added dropwise to a solution of 25 tnphenylphosphme (0 39 g, 1 50 mmol) and ethyl 2-[(10S)-3-hydroxy-10,l l-dihydro-5W-dibenzo[a,&lt;f]cyclohepten-10-yl]acetate (0.30 g, 1 00 mmol) in CH2C12 (5 mL) at 0 °C. The ice bath was removed and the reaction was allowed to warm to RT After 18 h, the solvent was removed under reduced pressure Flash chromatogrpahy (50% EtOAc/hexanes to 100% EtOAc, silica gel) gave 0 32 g of material that contained the desired product. A 30 second punfication by flash chromatography (75% to 90% EtOAc/hexanes, silica gel) gave 0.23 g of the desired material. MS(ES+) m/z 430.9 [M+H]*. <br><br> c) (S)-10,11 -dihydro-3-[3-(2-aminopyridin-4-yl)-1 -propyloxy]-5W-dibenzo[a,d]cycloheptene-10-acetic acid <br><br> 35 The compound of Example 1(b) (0 22 g, 0 50 mmol) was dissolved in IN NaOH <br><br> (0.77 mL, 0.77 mmol), EtOH (3 mL) and THF (3 mL). After heating the reaction at 50 °C for 18 h, the solvent was removed under reduced pressure The residue was dissolved in <br><br> -79- <br><br> Printed from Mimosa <br><br> WO 99/15508 PCT/US98/19466 <br><br> H,0 (4 mL) and filtered The filtrate was acidified with 30% TFA in H20 and the resulting precipitate was collected. Preparative HPLC (Hamilton PRP-1®, 3% CH3CN/H,0 - 0 1% TFA) gave 10 mg of the desired material as a hygroscopic solid MS(ES+) m/z 402 6 [M+Hf <br><br> 5 <br><br> Example 24 <br><br> Parenteral Dosage Unit Composition <br><br> 10 A preparation which contains 20 mg of the compound of Example 1 as a sterile dry powder is prepared as follows 20 mg of the compound is dissolved in 15 mL of distilled water The solution is filtered under sterile conditions into a 25 mL multi-dose ampoule and lyophilized The powder is reconstituted by addition of 20 mL of 5% dextrose in water (D5W) for intravenous or intramuscular injection. The dosage is thereby determined by the 15 injection volume Subsequent dilution may be made by addition of a metered volume of this dosage unit to another volume of D5W for injection, or a metered dose may be added to another mechanism for dispensing the drug, as in a bottle or bag for IV drip infusion or other injection-infusion system <br><br> 20 Example 25 <br><br> Oral Dosage Unit Composition <br><br> A capsule for oral administration is prepared by mixing and milling 50 mg of the 25 compound of Example 1 with 75 mg of lactose and 5 mg of magnesium stearate. The resulting powder is screened and filled into a hard gelatin capsule <br><br> Example 26 <br><br> 30 Oral Dosage Unit Composition <br><br> A tablet for oral administration is prepared by mixing and granulating 20 mg of sucrose, 150 mg of calcium sulfate dihydrate and 50 mg of the compound of Example 1 with a 10% gelatin solution The wet granules are screened, dried, mixed with 10 mg 35 starch, 5 mg talc and 3 mg stearic acid; and compressed into a tablet <br><br> -80- <br><br> Printed from Mimosa <br><br></p> </div>

Claims (28)

  1. <div class="application article clearfix printTableText" id="claims">
    <p lang="en">
    WO 99/15508<br><br>
    PCT/US98/19466<br><br>
    The above description fully discloses how to make and use the present invention However, the present invention is not limited to the particular embodiments described hereinabove, but includes all modifications thereof within the scope of the following claims. The various references to journals, patents and other publications which are cited herein comprises the state of the art and are incorporated herein by reference as though fully set forth.<br><br>
    -81 -<br><br>
    Printed from Mimosa<br><br>
    WO 99/15508 PCT/US98/19466<br><br>
    What is claimed is:<br><br>
    1 A compound according to formula (I)<br><br>
    wherein<br><br>
    A is CH2 or O,<br><br>
    R' is H, halo or Cj.galkyI, 10 R2 is H, C ] _6alkyl or CH2NR"R",<br><br>
    X is O or CH2,<br><br>
    Y is<br><br>
    N ^ _ NR"<br><br>
    R"R"hL .N<br><br>
    15<br><br>
    „NL .NR"-<br><br>
    N. ^NR"<br><br>
    N<br><br>
    R"R"N<br><br>
    or<br><br>
    G is NR", S or O,<br><br>
    R' is H, C].galkyl, OCi.galkyl, SC]-6alkyl, NR"R" or halo, 20 each R" independently is H or Cj.^alkyl, and s is 0, 1 or 2;<br><br>
    or a pharmaceutically acceptable salt thereof<br><br>
  2. 2 A compound according to claim 1 in which Y is<br><br>
    25<br><br>
    -82-<br><br>
    Printed from Mimosa<br><br>
    WO 99/15508<br><br>
    PCT/US98/19466<br><br>
    l\L ^NR"-<br><br>
    R'<br><br>
    , wherein R'is H, Cj^alkyl, OCj^alkyl,<br><br>
    SCj^alkyl, NR"R" or CI and each R" independently is H or Cj^alkyl.<br><br>
  3. 3 A compound according to claim 1 in which Y is R"R"N^<br><br>
    J<br><br>
    , wherein each R" is H or C|.4alkyl<br><br>
  4. 4 A compound according to claim 1 in which Y is<br><br>
    10<br><br>
    and s is 1<br><br>
    15<br><br>
    Cj.4alkyl.<br><br>
    ^nynr"<br><br>
    os-^nr"<br><br>
    , wherein each R" independently is H or C^alkyl,<br><br>
  5. 5 A compound according to claim 1 in which Y is<br><br>
    N<br><br>
    R"R"N —^ G<br><br>
    , wherein G is S and each R" independently is H or<br><br>
    20<br><br>
  6. 6 A compound according to claim 1 in which Y is<br><br>
    N NR"-<br><br>
    , wherein R" is H or C^alkyl.<br><br>
  7. 7 A compound according to claim 1 which is: (±)-10,11 -Dihydro-3-[2-(6-aminopyridin-2-yl)-1 -ethoxy]-5H-25 dibenzo[a,d]cycloheptene-10-acetic acid,<br><br>
    -83-<br><br>
    Printed from Mimosa<br><br>
    99/15508<br><br>
    PCT/US98/19466<br><br>
    (±)-10,11 -Dihydro-3-[4-(pyndin-2-ylamino)-1 -butyl]-5H-dibenzo[a,d]cycloheptene-10-acetic acid;<br><br>
    (±)-10,11 -Dihydro-3-[3-(4-ethoxypyridin-2-ylamino)-1 -propyloxy ]-5H-dibenzo[a,d]cycloheptene-10-acetic acid,<br><br>
    (S)-10,1 l-Dihydro-3-[3-(pyridin-2-ylamino)-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetic acid,<br><br>
    (R)-10,11 -Dihydro-3-[3-(pyridin-2-ylamino)-1 -propy loxy ]-5H-dibenzo[a,d]cycloheptene-10-acetic acid,<br><br>
    (±)-10,11 -Dihydro-3-[3-(3,4,5,6-tetrahydropyrimidin-2-ylamino)-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetic acid,<br><br>
    (±)-10,11 -Dihydro-3-[2-[2-(ethylamino)thiazol-4-yl]-1 -ethoxy]-5H-dibenzo[a,d]cycIoheptene-10-acetic acid;<br><br>
    (±)-10,11 -Dihydro-3-[3-(isoquinoline-1 -ylamino)-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetic acid;<br><br>
    (±)-10,11 -Dihydro-7-fluoro-3-[3-(pyndin-2-ylamino)-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetic acid;<br><br>
    (S)-10,1 l-Dihydro-3-[3-(4-methylpyndin-2-ylamino)-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetic acid;<br><br>
    (S)-10,11 -Dihydro-3-[3-(4-ethoxypyndin-2-ylamino)-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene- 10-acetic acid;<br><br>
    (±)-10,1 l-Dihydro-6-methyl-3-[3-(pyndin-2-ylamino)- l-propyloxy]-5H-dibenzo[a,d]cycloheptene-l 0-acetic acid;<br><br>
    (±)-10,ll-Dihydro-2-(dimethylamino)methyl-7-fluoro-3-[3-(pyridin-2-ylamino)-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-l 0-acetic acid,<br><br>
    (S)-10,ll-Dihydro-3-[3-[4-(2-propyloxy)pyndin-2-yIamino]-l-propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetic acid;<br><br>
    (S)-10,ll-Dihydro-3-[2-[6-(methylamino)pyridin-2-yl]-l-ethoxy]-5H-dibenzo[a,d]cycloheptene-10-acetic acid;<br><br>
    (S)-10,11 -Dihy dro-3-[3- [4-(dimethy lamino)pyridin-2-ylamino]-1 -propy loxy]-5H-dibenzo[a,d]cycloheptene-10-acetic acid;<br><br>
    (+)-10,11 -Dihydro-3-[3-[4-(ethylthio)pyridin-2-ylamino]-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetic acid,<br><br>
    (S)-10,11 -Dihydro-3-[3-(4-chloropyndin-2-y lamino)-1 -propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetic acid,<br><br>
    (±)-10,11 -Dihydro-2-methy 1-3- [3-(pyndin-2-y lamino)-1 -propy loxy ]-5H-dibenzo[a,d]cycloheptene-10-acetic acid,<br><br>
    -84-<br><br>
    Printed from Mimosa<br><br>
    (S)-10,1 l-Dihydro3-[3-(4-aminopyridin-2-ylamino)-l -propyloxy]-5H-dibenzo[a,d]cycloheptene-10-acetic acid;<br><br>
    (±)-10,11 -Dihydro-3-[3-(4-methy lpyridm-2-ylamino)-1 -propyloxy ]-dibenzo[b,f]oxepine-10-acetic acid;<br><br>
    (±)-10,11 -Dihydro-3-[2-[6-(methylamino)pyridm-2-yl]-1 -ethoxy]-dibenzo[b,f]oxepme-10-acetic acid; or<br><br>
    (S)-10,11 -Dihydro-3- [3-(2-aminopyridin-4-yl)-1 -propy loxy ]-5H-dibenzo[a,c/]cycloheptene-10-acetic acid;<br><br>
    or a pharmaceutically acceptable salt thereof.<br><br>
  8. 8: A pharmaceutical composition which comprises a compound according to any one of claims 1 to 7 and a pharmaceutically acceptable carrier.<br><br>
  9. 9. A pharmaceutical composition which comprises a compound according to any one of claims 1 to 7, an antineoplastic agent and a pharmaceutically acceptable carrier.<br><br>
  10. 10. The pharmaceutical composition according to claim 9 wherein the antineoplastic agent is topotecan.<br><br>
  11. 11. The pharmaceutical composition according to claim 9 wherein the antineoplastic agent is cisplatin.<br><br>
  12. 12. A compound according to formula (II);<br><br>
    A is CH2 or O;<br><br>
    R1 is H, halo or Cj.galkyl; R2 is H, C]-6alkyl or CH2NR"R"; X is O or CH2;<br><br>
    Y is y<br><br>
    COgC^alkyl<br><br>
    (II)<br><br>
    wherein:<br><br>
    '/NTELLECTUAL PROPERTY OFFICE OF N.Z.<br><br>
    -85-<br><br>
    ' 2 DEC 2001 received<br><br>
    n<br><br>
    NR"-<br><br>
    n<br><br>
    NR"<br><br>
    R'<br><br>
    R"R"N<br><br>
    or<br><br>
    H<br><br>
    G is NR", S or O;<br><br>
    R' is H, Q.^alkyl, OC]-6alkyl, SCj^alkyl, NR"R" or halo; each R" independently is H or Cj.galkyl; and s is 0, 1 or 2;<br><br>
    or a pharmaceutically acceptable salt thereof.<br><br>
  13. 13. A compound according to formula (III):<br><br>
    A is CH2 or O;<br><br>
    R1 is H, halo or C^galkyl,<br><br>
    R2 is H, Ci_6alkyl orCH2NR"R";<br><br>
    X is O or CH2;<br><br>
    R' is H, Cj^alkyl, OCi-6alkyI, SCi-6alkyl, NR"R" or halo; and each R" independently is H or Cj.^alkyl;<br><br>
    or a pharmaceutically acceptable salt thereof<br><br>
  14. 14. A process for preparing a compound of the formula (I) as defined in claim 1, which process compnses reacting a compound of formula (IV) with a compound of formula (V).<br><br>
    R'<br><br>
    (HI)<br><br>
    wherein:<br><br>
    -86-<br><br>
    1 2 DEC 2001 received<br><br>
    C02C^alkyl Y-(CH2)2_3-Ll<br><br>
    (IV) (V)<br><br>
    wherein R*, R2, Y and A are as defined in claim 1, with any reactive functional groups protected, and is OH or halo,;and thereafter removing any protecting groups, and optionally forming a pharmaceutically acceptable salt.;15. A process for preparing a compound of the formula (I) as defined in claim 1, which process comprises reacting a compound of formula (IV) with a compound of 10 formula (VI).;C02Ci-alkyl;(IV);N* NR" (CH2)2, —OH<br><br>
    R'<br><br>
    (VI)<br><br>
  15. 15 wherein R^, R^, R\ R" and A are as defined m claim 1, with any reactive functional groups protected,<br><br>
    and thereafter removing any protecting groups, and optionally forming a pharmaceutically acceptable salt.<br><br>
    20
  16. 16. A process for preparing a compound of the formula (I) as defined in claim<br><br>
    1, which process comprises reacting a compound of formula (IV) with a compound of formula (VII).<br><br>
    25<br><br>
    (IV)<br><br>
    C02C1 „alkyl o-<br><br>
    R\<br><br>
    \<br><br>
    ,N _N+ ^(CH2)2.3—OH<br><br>
    (VII)<br><br>
    -87-<br><br>
    INTELLECTUAL PROPERTY OFFICE OF N.Z.<br><br>
    1 2 DEC 2001 received<br><br>
    wherein R1, R2, R" and A are as defined in claim 1, with any reactive functional groups protected;<br><br>
    and thereafter removing any protecting groups, and optionally forming a pharmaceutically acceptable salt.<br><br>
  17. 17. A compound according to any one of claims 1 to 7 for use as a medicament.<br><br>
  18. 18. The use of a compound according to any one of claims 1 to 7 in the manufacture of a medicament for the treatment of diseases in which antagonism of the avP3receptor is indicated.<br><br>
  19. 19. The use of a compound according to any one of claims 1 to 7 in the manufacture of a medicament for the treatment of diseases in which antagonism of the a^receptor is indicated.<br><br>
    i<br><br>
  20. 20. The use of a compound according to any one of claims 1 to 7 in the manufacture of a medicament for the treatment of osteoporosis.<br><br>
  21. 21. The use of a compound according to any one of claims 1 to 7 in the manufacture of a medicament for the inhibition of angiogenesis.<br><br>
  22. 22. The use of a compound according to any one of claims 1 to 7 in the manufacture of a medicament for the inhibition of tumor growth or tumor metastasis.<br><br>
  23. 23. The use of a compound according to any one of claims 1 to 7 in the manufacture of a medicament for the treatment of atherosclerosis or restenosis.<br><br>
  24. 24. The use of a compound according to any one of claims 1 to 7 in the manufacture of a medicament for the treatment of inflammation.<br><br>
  25. 25. The use of a compound according to any one of claims 1 to 7 and an antineoplastic agent in the manufacture of a medicament for the inhibition of tumor growth in physical combination or for stepwise administration<br><br>
    -88-<br><br>
    INTELLECTUAL PROPERTY OFFICE OF NZ.<br><br>
    1 2 DEC 2001 received<br><br>
  26. 26. The use according to claim 25 wherein the antineoplastic agent is topotecan.<br><br>
  27. 27. The use according to claim 25 wherein the antineoplastic agent is cisplatin.<br><br>
  28. 28. The use of a compound according to any one of claims 1 to 7 and an inhibitor of bone resorption in the manufacture of a medicament for the treatment of osteoporosis in physical combination or for stepwise administration.<br><br>
    10<br><br>
    15<br><br>
    20<br><br>
    25<br><br>
    30<br><br>
    35<br><br>
    intellectual property office of n.z.<br><br>
    1 2 DEC 2001<br><br>
    received<br><br>
    -89-<br><br>
    </p>
    </div>
NZ503389A 1997-09-19 1998-09-18 Vitronectin receptor antagonists comprising substituted dibenzo[a,d]cycloheptene or dibenzo[b,f]oxepine and their use as antagonists of alpha-v-beta-3 receptors or alpha-v-beta-5 receptors NZ503389A (en)

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Families Citing this family (18)

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Publication number Priority date Publication date Assignee Title
CO4920232A1 (en) * 1997-01-08 2000-05-29 Smithkline Beecham Corp DIBENZO [A, D] CYCLLOHEPTANE ACETIC ACIDS WITH VITRONECTIN RECEPTOR ANTAGONIST ACTIVITY
WO2000046215A1 (en) 1999-02-03 2000-08-10 Merck & Co., Inc. Benzazepine derivatives as alpha-v integrin receptor antagonists
DE19936780A1 (en) 1999-08-09 2001-02-15 Basf Ag New integrin receptor antagonists
US6881736B1 (en) 1999-09-07 2005-04-19 Smithkline Beecham Corporation Vitronectin receptor antagonists
EG24179A (en) * 1999-09-07 2008-09-28 Smithkline Beecham Corp Vitronectin receptor antagonists
US6514964B1 (en) 1999-09-27 2003-02-04 Amgen Inc. Fused cycloheptane and fused azacycloheptane compounds and their methods of use
FR2806082B1 (en) * 2000-03-07 2002-05-17 Adir NOVEL BICYCLIC ANTAGONIST VITRONECTIN RECEPTOR COMPOUNDS, PROCESS FOR THEIR PREPARATION AND PHARMACEUTICAL COMPOSITIONS CONTAINING THEM
EP1289959A1 (en) 2000-06-15 2003-03-12 Pharmacia Corporation Dihydrostilbene alkanoic acid derivatives useful as vitronectin antagonists
JP2004513953A (en) * 2000-10-24 2004-05-13 メルク　エンド　カムパニー　インコーポレーテッド Dibenzoxazepine αV integrin receptor antagonist
CN1247258C (en) 2001-04-24 2006-03-29 默克专利有限公司 Combination therapy using anti-angiogenic agents and tnfa
GB0215867D0 (en) * 2002-07-09 2002-08-14 Glaxosmithkline Spa Novel method and compounds
US20040224986A1 (en) 2002-08-16 2004-11-11 Bart De Corte Piperidinyl targeting compounds that selectively bind integrins
UA87854C2 (en) 2004-06-07 2009-08-25 Мерк Энд Ко., Инк. N-(2-benzyl)-2-phenylbutanamides as androgen receptor modulators
EP1973569B1 (en) 2006-01-18 2013-05-22 Merck Patent GmbH Specific therapy using integrin ligands for treating cancer
US20100069302A1 (en) 2007-07-18 2010-03-18 Stefan Krueger Specific therapy and medicament using integrin ligands for treating cancer
JP2012528079A (en) 2009-05-25 2012-11-12 メルク パテント ゲゼルシャフト ミット ベシュレンクテル ハフツング Continuous administration of integrin ligand to treat cancer
KR20160147007A (en) 2014-05-30 2016-12-21 화이자 인코포레이티드 Carbonitrile derivatives as selective androgen receptor modulators
WO2023275715A1 (en) 2021-06-30 2023-01-05 Pfizer Inc. Metabolites of selective androgen receptor modulators

Family Cites Families (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CH443281A (en) * 1964-02-18 1967-09-15 Hoffmann La Roche Process for the preparation of dibenzocycloheptatriene compounds
US3335148A (en) * 1966-02-17 1967-08-08 Lilly Co Eli 9(3-pyridyl)derivative of fluorene, 9-fluorenol, xanthene, 9-xanthenol and the corresponding nonphytotoxic acid addition salts thereof
US5698551A (en) * 1995-04-07 1997-12-16 Novo Nordisk A/S Heterocyclic compounds
JP3960482B2 (en) * 1995-06-29 2007-08-15 スミスクライン・ビーチャム・コーポレイション Integrin receptor antagonist
US5659033A (en) * 1995-09-13 1997-08-19 Neurogen Corporation N-aminoalkylfluorenecarboxamides; a new class of dopamine receptor subtype specific ligands
WO1998015278A1 (en) * 1996-10-07 1998-04-16 Smithkline Beecham Corporation Method for stimulating bone formation
CO4920232A1 (en) * 1997-01-08 2000-05-29 Smithkline Beecham Corp DIBENZO [A, D] CYCLLOHEPTANE ACETIC ACIDS WITH VITRONECTIN RECEPTOR ANTAGONIST ACTIVITY

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