CA2174650A1 - Piperidinyl camphorsulfonyl oxytocin antagonists - Google Patents

Abstract

Spirocyclic compounds of formula (I), where m is an integer from zero to one; Y is CH or CH2; and X is C=O, CH2, PO2H, PO2Z, where Z is C1-10 alkyl, SO2, CONH, or SO2NH. Such compounds are useful as oxytocin and vasopressin receptor antagonists useful in the treatment of preterm labor, dysmenorrhea and for the stoppage of labor preparatory to cesarean delivery, timing of parturition, uterine hyperactivity, endometriosis, hypertension, congestive heart failure, hyponatremia and cognitive disorders. Also disclosed are pharmaceutical compositions containing these compounds, methods of their use and methods of their preparation.

Description

2~ 21 74 65~ PCT/IJS94/13483 TITLE OF THE INVENTION
PLPERIDINYLCAMPHORSULFONYL OXYTOCIN ANTAGONISTS
FIELD OF THE INVENTION
This application is a cnntin~ tinn-in-part of U.S. Seriai No.
08/153,521, filed November 16, 1993, the contents of which are hereby incorporated by reference, which is a continll ~rinn-in-part of U.S. Serial No. 07/954,596, filed September 30, 1992, now abandoned, which is a cul,lillu~ion of U.S. Serial No. 07/759,254, filed September 13, 1991, now abandoned, which is a cnn~in~ ion-in-part of U.S. Serial No.
07/612,344, filed November 13, 1990, now ~h~n~lnnP~I the contents of all previous app]ications are hereby incorporated by reference.
The present imvention provides novel compounds, novel compositions, methods of their use and methods of their m~nllf~chlre7 such compounds generally pharmacologically useful as agents in obstetric and gynecologic therapy. The aforementioned phammacologic activities are useful in the treatment of m~mm~l~ More specifically, the compounds of the present invention can be used in the treatment of preterm labor, stopping labor preparatory to Caesarean delivery, and in the treatment of dysmenorrhea. At the present time, there is a need in the area of obstetric and gynecologic therapy for such agents.
BACKGROUND OF THE INVENTION
In the field of obstetrics, one of the most important problems is the management of preterm labor. A significant number of the pregnancies progressing past 20 weeks of gestation experience p~Clll~Ult~ labor and delivery, which is a leading cause of neonatal morbidity and mortality. Despite major advances in neonatal care, retention of the fetus in utero is preferred in most instances.
Tocolytic (uterine-relaxing) agents that are currently in use include ~2-adrenergic agonists, m~nPsillm sulfate and ethanol.
Ritodrine, the leading ~2-adrenergic agonist, causes a number of cardiovascular and metabolic side effects in the mother, including tachycardia, increased renin secretion, hyperglycemia (and reactive .... , . . . , .. .. . . ... . _ _ _ _ _ _ _ _ _ WO 95/14025 PCr~US94113483 21 7~650 hypoglycemia in the infant). Other ~2-adrenergic agonists, including terbutaline and albuterol have side effects similar to those of ritodrine.
nf~Sillm sulfate at plasma concentrations above the therapeutic range of 4 to 8 mg/dL can cause inhibition of cardiac conduction and 5 neuromuscular transmission, ~ dloly depression and cardiac arrest, thus malcing this agent unsuitable when renal function is impaired.
Ethanol is as effect;ve as ritodrine in preventing pl~lllalulc labor, but it does not produce a corresponding reduction in the incidence of fetal d~ y distress that administration of ritodnne does.
It has been proposed that a selective oxytocin antagonist would be the ideal tl~colytic agent. In the last few years, evidence has ~rcllm~ tPd to strongly suggest that the hormone oxytocin may be a physiological initiat~r of labor in several m~mm~ n species including humans. Oxytocin is believed to exert this effect in part by directly 5 contracting the uterine myometrium and in part by ~nh~nrin~ the synthesis and release of contractile pr )st~ n-lins from the uterine endometrium/decid~a. These pr st~ n~lins may, in addition, be illll,ollallL in the cer~/ical ripening process. By these ml~l h~nicms, the process of labor (term and preterm) is initiated by a heightened 20 sensitivity of the ut~rus to oxytocin, resulting in part as a result of a well-documented increase in the number of oxytocin receptors in this tissue. This "up-regulation" of oxytocin receptors and enhanced uterine sensitivity appears to be due to trophic effects of rising plasma levels of estrogen towards term. By blocking oxytocin, one would block both the 25 direct (contractile) and indirect (enhanced prostaglandin synthesis) effects of oxytocin on the uterus. A selective oxytocin blocker, or antagonist, would lilcely be more efficacious for treating preterm labor than current regimens. In addition, since oxytocin at term has major effects only on the uterus, such an oxytocin antagonizing compound 30 would be expected t~ have few, if any, side effects.
The compounds of the present invention are also useful in the treatment of dysmenorrhea. Thi~ condition is characteri~ed by cyclic pain associated with menses during ovulatory cycles. The pain i.s thought to result from uterine contractions and ischemia, probably WO 95/14025 2 1 7 4 6 5 0 Pcr~Sg4/l3483

- 3 -mediated by the effect of prost~ nrlins produced in the secretory endometrium. By blocking both the direct and indirect effects of oxytocin on the uterus, a selective oxytocin antagonist can be more efficacious for treating dy~ eno~lllea than current regimens.
An additional use for the present invention is for the stoppage of labor ~ ,al~lluly to Caesarean delivery. Certain spiroindanylpiperidines and spiroind~llyl~ilJelidines are known (U.S.
Patents 3,654,287 and 3,666,764), however, they are reported to be useful as ~n~sth!~ti~ agents which is quite distinct from the utility of the o present invention.
It is, therefore, a purpose of this invention to provide sllbst:lnces which more effectively antagonize the function of oxytocin in disease states in animals, preferably m~mm~l~, especially in humans. It is another purpose of this invention to prepare novel compounds which more selectively inhibit oxytocin. It is still another purpose of this invention to provide a method of antagonizing the functions of oxytocin in disease states in m~mm:~lc It is also a purpose of this invention to develop a method of preventing or treating oxytocin-related disorders of preterm labor and dy~lllenullllea by antagonizing oxytocin.
It has now been found that compounds of the present invention are antagonists of oxytocin and bind to the oxytocin receptor.
When the oxytocin receptor is bound by the compounds of the present invention, oxytocin is antagonized by being blocked from its receptor and thus being unable to exert its biologic or pharmacologic effects.
25 These compounds are useful in the treatment and prevention of oxytocin-related disorders of animals, preferably m~mm~ls and especially humans. These disorders are primarily preterm labor and dysmenorrhea. The compounds would also find usefulness for stoppage of labor preparatory to Caesarean delivery. Additionally, such 30 compounds are useful in inducing cu-l~ldc~plion in m:~mm~l~ inasmuch a~ oxytocin antagonists have now been shown to inhibit the release of oxytocin-stimulated llltPini~in~ hormone (LH) by anterior pituitary cells.

2 1 7~

- 4 -Compounds of the present invention are also inhibitors of vasopressin and can bind to the vasopressin receptor. These compounds are useful in inducillg vasodilation, treating hypertension, inducing diuresis and inhibiting platelet ~ tin~fion.
Additionally, it has now been found that the compounds of the present inventioll are balanced oxytocin and vasopressin antagonists useful for treating preterm labor and dysmenorrhea, SU~IARY OF TH~ NTION
0 The compounds and their pharrn~( e--fic~lly acceptable salt.s and esters of the present invention are those of the general structural formula I
~2 R3 ~\~ Rs ,R6 ~ R Y
L~ R8 wherem X is o C
3 o CH2.
P02H, P02Z where Z is Cl lo alkyl, so2~
S02NH or -

5 ~ PCrlUS94/13483 o C - NH .
Y is absent or is CH or CH2;
m is an integer of from zero to one;
o n is an integer of from zero to five;
p is an integer of from zero to five;
q is an integer of from zero to five;
Rl and R2 are each independently hydrogen, halogen, hydroxy, Cl -10 alkyl, Cl lo alkoxy or trifluoromethyl;
R3 is hydrogen, halogen, hydroxy or oxo with the proviso that when R3 is oxo then the 2,3 bond is saturated;
R4is 3 0 hydrOgell, phenyl, or Cl lo alkyl, R5 and R6 are in-l~pçn~ ntly hy ~roge ., WO 95/14025 Pcr/uss4ll34s3 .
- - 21 1~6

- 6 -C1 1o alkyl or Cl 1o hydroxyalkyl; or RS and R6 together are oxo or n~nbstitllted or substituted C3-6 cycloalkyl, where said substituents are hydroxy, Cl lo alkyl, CI -10 hydroxyalkyl, Cl lo alkoxy or Cl-3 alkoxyalkoxyalkoxyalkyl;
R7and R8 are independently hydrogen or hydroxyl;
R9 is C7 1 0 alkoxy, Cl -10 alkoxycarbonyl, Cl l o alkoxycarbonylalkylarninocarbonyl, cyano, --O--P-OH
OH
Cl lo alkyl substituted phosphonate, --O-S- OH

Cl lo alkyl substituted sulfonate, --(CH2)n --N--R~2 W0 95/14025 2 1 7 4 ~ ~ ~ PCrlUss4/l34s3 --(CH2)p --N- CO--Rl~
--(CH2)q--CO-NH--R14 or llnsllbstihl~d or substituted C1 l0 alkyl where said lr ~ ~ l is Rl; or R7 and R8 are, together with the carbons to which they are attached, o joined to form a 5-membered hetercyclic ring containing 2 hetero atoms where said hetero atoms are N and O;
R10 is hydroxyl, carboxyl, C1 loalkoxy, C1 1o alkoxycarbonyl, R1S or cyano;

Rll iS
hydrogen, C 1- l o alkyl, Cl lo carboxyalkyl or C1 1o alkoxycarbonylalkyl;
R12is hydrogen, Cl lo alkylsulfonyl, Cl_10 alkarylsulfonyl, C1-10 aralkylsulfonyl, Cl 1o alkoxyarylsulfonyl, aminosulfonyl, Cl lo alkylaminosulfonyl, Cl lo dialkylaminosulfonyl, llncllhstihlt~d or sllh~stihltpd C4 15 cycloalkylalkyl, bicycloalkyl or tricycloalkyl where said substituent is oxo or sulfonyl sllh.s~ih~tPd by uul~ub~Lilul~d or s~lhstih-t~d C7 ls cycloalkyl, bicycloalkyl or tricycloalkyl where said ,lr~ are oxo, o~ime or hydroxy;
R13 is amino, Cl lo alkylarnino, Cl lo alkyl which is llnsllhstihltPd or mono- or di-substituted by 6, C2 l0 alkeny~ which is unsubstituted or substituted by Rl6, phenyl substituted by R 17, lln~lhctitlltPd or sllh~tihlt~d C3-8 cycloalkyl where said sllhstitllPnt.~ al-e Cl lo ~Ikyl or carboxy, N riic--hstitlltPd by Cl lo alkyl, llnsllhstihltPd or suhstih-tPd C7 l5 bicycloalkyl or tricycloalkyl where said s--l-~tit-~Pnt i.s carbox~, Cl lo alkoxy or R18;
Rl4 i.s Cl-lO alkyl, Cl lo aminoalkyl, Cl lo alkoxycarbonylalkyl, or C l l o carboxyalkyl, _ 9 _ R15 is lm~lhstihltl-d or SllhstitlltPd 5 membered heterocyclic rings containirlg I hetero atom where said hetero atom is N and said substituents are 0X0, amino, Cl 1o alkylamino, Cl 1o carboxyalkylcarbonylamino, Cl lo dicarboxyalkylamino or o Cl lo alkyloxycarbonylalkylamino;
R16 is llr~cllhstihlt~d or s~lbstih~t~d C4-8 cycloalkyl where said substituents are hydroxy or carboxy, C10 15 bi- or tricycloalkyl, halogen, hydroxy, carboxy, oxo, oxime, Cl lo aikylthio, Cl lo alkylsulfunyl, Cl_10 alkylsulfonyl, Cl lo alkoxycarbonyl, R20, R21, amino, aminocarbonyl, Cl lo dialkylaminocarbonyl, Cl lo alkylamino, Cl lo dialkyiamino, Cl lo alkylcarbamate, WO 95/14025 PCrlUS94113483 .
~7~5 Cl lo alkylcarbonate, C1 loalkylureide, Cl lo aralkyl.;a-l,all.dl~, lm~llh~titlltl~d or sl~hstihlt~d aryloxy where said substituents are amino, C1 1o alkyl or Cl lo aminoalkyl, Cl lo aralkoxy, n~llhStihlt~d or ,~llhstihlf~d Cl 1o alkaryloxy where said sllhstihl.ont is Cl lo ~Ikylcarbamate, N di~ub~Lilu~d by Cl lo alkyl and Cl lo carboxyalkyl or N tri-substituted by two Cl lo alkyls and by Cl lo alkoxycarbonylalkyl with the proviso that a trifluoroacetic acid counterion be present;
Rl7 is amino, 2 halogen, Cl_lo alkyl, Cl loalkoxy, nitro, phenylcarbon~l or un~ub~LiLul~d ~r ~l-bctihlt~d 5 membered heterocyclic rings c-nt~inin~ I hetero atom where said hetero atom is O and wherein said ~ub~Li~u~ is oxo;
R 18 is ull~ul~liLul~d or ~lhstihll~d heterocyclic rings selected from azetidinyl, pyrrolidinyl, pyrrolyl, piperidinyl, ;

WID 9~114025 2 1 7 4 6 5 0 PCrlUS94/13483 piperizinyl, pyridinyl, pyrimidinyl, tetrahydrofuranyl, furanyl7 dioxolanyl, thienyl, 1, 3-thiazolidinyl or tetrahydrooxazolyl; where said s~lhstit-~PnS~ are one or more of 1 o oxo, hydroxy, carboxy, amino, Cl lo carboxyalkyl, Cl_10 alkyl, Cl 1oalkoxy, C1 -10 aralkoxy, C1 1oalkaryloxy, C1 1o alkoxycarbonyl, Cl-10 alkoxycarbollylamino~
CI-10 alkoxycarbonylalkyl, Cl lo aralkoxycarbonyl or s-lh~titl-tPd or lln~llhstitlltPd phenyl where said sub~Lilu~
are 2~ Cl_5 alkyl, carboxy or halogen;
R20 is lln~lhstitlltPd or substituted heterocyclic rings selected from azetidinyl pyrrolidinyl, pyrrolyl, tetrahydroimidazolyl, irnidazolyl, WO 9S114025 PCI'IUS94/13483 2~ 7~65~

tetrazolyl, piperidinyl, pyridinyl, hexah~dloa~ yl, thienYI~
1, 3-thiazolidinyl or tetrahydrothi~lzinyl; where said sllhsfitll~ntc are one or more of C1-10 alkyl, o Cl-10 aralkyl.
Cl lo aralkoxy, Cl lo alkaryl, amino, C1 1o alkylarnino, Cl -10 dialkylarnino, oxo, oxime, fused phenyl, C1 1o alkoxycarbonyl, C1 1o ;llkylcarbonate, C1 lo alkylureide, Cl lo alkyl~,dll)al.laL~, or llncllhs~itllt~-l or substituted ~-membered heterocyclic rings ha~ing I hetero atom where said hetero atom is N and said s~lhstitl~nt is one or more of oxo or fused phenyl;
R21 iS
unsub~tituted or s~hstit--~d phenyl where said substituent~ are halogen, C1 1o alkyl, C I 10 carboxyalkyl, Cl lo alkoxy, r WO9S/14025 2~ 74~ PCI/US94113483 5- or 6-membered heterocyclic rings having l or 2 hetero atoms where said hetero atoms are N or S, hydro~y, carbo~y or --O-S--OH
o In one embodiment of the instant invention are compounds the formula ~, X~y~

~vherein Xis o C, CH2, Po2H7 PO2Z ~vhere Z is C1 l0 alkyl, or SO2;

Rl and R2 are each indPp~n~l~n~ly hydrogen, halogen or Cl lo alkyl;

WO 95/14025 PC~IUS94/13483 21 7~65G

R4 is hydrogen or phenyl;
RS and R6 are independently hydrogen or C1 1o alkyl; or RS and R6 together are oxo or n~-hstihlt~d or ~llhctihltt~d C3-6 cycloalkyl, where said sllhstit-l.onts are hydroxy, CI-10 hydroxyalkyl or C1-3 al~oxyalkoxyalkoxyalkyl;
R9 is C7-1o alkoxy, C1 1o alkoxycarbonyl, C1 1o alkoxycarbonylalkylarninocarbonyl, --O--P-OH
C1 1o alkyl substituted OH
ll o phosphonate, Cl lo alkyl substituted sulfonate, --(CH2)n --N--R12 --(CH2)p --1\1- CO--Rl3 llnsllh~titllt~d or substituted Cl lo alkyl where said substituent i.s WO 95/14025 2 1 7 ~ ~ 5 ~ PCT/IJS94113483 R10; or R7 and R8 are, together with the carbons to which they are attached, joined to form a 5-membered heterocyclic ring containing 2 hetero 5 atoms where said hetero atoms are N and O;
R12 iS
hydrogen, Cl loalkylsulfonyl, o C1 1oalkarylsulfonyl, Cl lo alkoxyarylsulfonyl, ~min--sulfonyl, C1 1odialkylaminosulfonyl, ns~lh~titlltPd or sllbstit~lt~ d C3-15 cycloalkylalkyl where said 1 5 .s. .~ iS
oxo or sulfonyl sl~hstihlt~d by IIII~III).,Iill~l~d or substituted C3 15 cycloalkyl where said ~ ll is oxo R 14 is Cl lo alkyl, Cl lo arninoalkyl or Cl loalkoxycarbonylalkyl;
R16 iS
lln~llhctihltt~d or ~llhstitllt~d C5-6 cycloaLkyl where said s~lhstitll~nt is hydroxy, C10-15 tricycloalkyl7 halogen, hydroxy, carboxy, oxo, WO 9~/1402~ PCTIUS94113483 2~ 74650 Cl loalkyltlhio, C1 lo alkylsulfonyl, C1 lo alkoxycarbonyl, R20, R21, amino, Cl -10 alkylamino, C1 1o dialkylamino, Cl lo alkylc~lb~ulla~
Cl loaralkyl~all,~"là~, unsubstituted or ~ub~Li~ul~d aryloxy where said sllh~tit~ nt~
are arnino, Cl 1o alkyl or C1 1oaminoalkyl, C1 1o aralkoxy, lln~llhstitlltPd or sllhstitllt~d C1 l0 alkaryloxy where said l l r l l l is Cl lo alkyl ,alballlat~, N ,li~ d by Cl lo alkyl and Cl lo carboxyalkyl; or N tri-s--hstitllt~d by two Cl-lo alkyls and by Cl-lO
alkoxycarborlylalkyl with the proviso that a counterion be present from tlhe group c--n~i~tin~ of Cl ~ halogenated carboxylic acids;

Rl~ is llncllh~titllt~d or s--h~tin-t~d II~Lelu~j~lic rings selected from azetidinyl, pyrrolidinyl, pyrrolyl, plpendmyl, piperizinyl, pyridinyl, pyrimidinyl, tetrahydrofuranyl, WO 95/14025 2 ~ 7 ~ 6 5 0 Pcr/usg4/l3483 .

furanyl, dioxolanyl, thienyl, 1, 3-thiazolidinyl or tetrahydrooxazolyl, where said substituents are one or more of oxo, hydroxy, carboxy, Cl lo carboxyalkyl, o C1 loalkyl, C1 1o alkoxy, Cl lo aralkoxy, C1 1oalkoxycarbonyl, Cl lo alkoxycarbonylalkyl or C1 1o aralkoxycarbonyl;
R20 is l~n~llhstinl~Pd or substituted heterocyclic rings selected from hexahydroazepinyl, pyrrolidinyl, pyrrolyl, tetrahydroimidazolyl, imidazolyl, tetrazolyl, piperidinyl, pyridinyl, azetidinyl, thienyl, 1, 3-thiazolidinyl or tetrahydrothiazinyl; where said ~llb~titl-~nt~ are one or more of Cl-lO alkyl, Cl lo aralkyl, Cl lo aralkoxy, arnino, ~.~

WO 9S/14~nS PCrlUS94113483 fused phenyl, Cl 1o alkoxycarbonyl, Cl lo alkylcarbamate, or lm~llh~titlltt-d or s~lhstitlltPd S-membered heterocyclic rings having I hetero atom where said hetero atom is N and said Ut;lll iS
oxo or fused phenyl;
10 R21 iS
unsubstituted or substituted phenyl ~vhere said substituent~ are Cl-10 alkyl, Cl la carboxyalkyl, Cl lo alkoxy, S- or 6-membered heterocyclic rings having I or 2 hetero atoms where said hetero atoms are N or S, hydroxy, carboxy or o --O-S--OH
a In a class are the compounds selected from WO95/14025 21 74 650 PC!rluss4ll34s3 .

CH3 ,CH3 N : ~
o ~CH3 HN ~H3 So CH3 [~ CH3 ,CH3 :~o N
2s ~H
HN~So CH3 In another class are the compounds selected from WO 9511402!i PCrrllS94/13483 2 1~65~ --~2~?oH o O NHz N ~ ;~
~\ or ~OH
S2 1l H--~- ?
H
Illustral:ive of the invention is a pharmaceutical composition, Cu~ g a compound of formula I in a pharmacologically effective amount for antagonizing the binding of oxytocin to its receptor site, and a ph~-l-la~u~ically acceptable carrier.
Illustrations of the present invention include methods of antagonizing the binding of oxytocin to its receptor site, preventing preterrn labor, treating dysmenorrhea, and stopping labor preparator~
to cesarean delivery, in a mammal in need thereof, comprising the ~tep of ~.l,,,i,,;~lP, jllg to the mammal a pharmacologically effective amount of a compound of formula I.
Exemplifying the invention are method~ of antagonizing vasopressin from binding to it~ receptor site, inducing vasodilation, treating hypertension, inducing diuresi~. and inhibiting platelet :~g~ tin~tion, in a mammal in need thereof comprising the step of wo 95/14025 2 1 7 4 ~ ~ ~ PC~rllJS94113483 rl illg to the mammal a pharmacologically effective amount of a compound of formula 1.
An example of the invention is a method of increasing fertility and embryonic survival in a farm animal comprising 5 adl~ elillg to the farm animal a pharmacologically effective amount of a compound of the present invention.
Further illustrating the invention is a method for improving survival of a farm animal neonate CUIIIIJI i~illg controlling timing of parturition to effect delivery of the neonate during daylight hours by adlllilli~l~lillg to a farm animal which is expected to deliver the neonate within 24 hours a pharmacologically effective amount of a compound of the present invention.
Another illustration of the invention is a method of controlling the timing of estrus in a farm animal, comprising 15 ~-lminist~ring to the farm animal a pharmacologically effective amount of a compound of the present invention.
Also included in the instant invention is a compound of the formula n HN
n `loT ~ ~
H
and the pharm~relltir~lly acceptable salt.s thereof.
An embodiment of the invention is a ph~rm~elltical composition, comprising the compound of formula n in a . .

WO 95/1402~ PCTIUS94/13483 21 7~55 pharmacologically effective amount for antagonizing the binding of oxytocin to its receptor site, and a pharm~cel-tir ~lly acceptable carrier.
Illustrative of the invention are methods of antagonizing the binding of oxytocin to its receptor site, preventing preterm labor, 5 treating dysmrnnrrhp~ and stopping labor preparatory to cesarean delivery, in a mammal in need thereof, comprising the step of ad.~ li..g to the mammal a pharmacologically effective amount of a compound of formula II.
Also ~nmp~ed in the inst~nt invention are methods of treating preterm labor and d~ lellolll-ea in a mammal in need thereof, c~,lll~,lisillg ~ rl;ll~ to the mammal a pharmacologically effective amount of a compoumd which binds to a human oxytocin receptor with a binding affinity which is no more than ten-fold higher or ten-fold lower than the binding affinity with which the compound binds to a human 15 arginine-vasopressill-vla (AVP-Vla) receptor.
More particularly illustrating the the invention are the methods of treating preterm labor and dysmenorrhea, wherein the compound binds to the human oxytocin receptor with a binding affinity which is no more than five-fold higher or five-fold lower than the 20 binding affinity with which the compound binds to the human AVP-VIa receptor.
Exemplifying the invention are the methods of treating preterrn labor and dysmenorrhea, wherein the compound is selected from ~2 -- OH o NH2 NH J~
or WO 95/14025 2 1 7 4 6 5 ~ PCI/US94/13483 ~2 Salts and esters encompassed within the term "ph:lrm~relltir~lly acceptable salts and esters" refer to non-toxic salts of the compounds of this invention which are generally prepared by reacting the free base with a suitable organic or inorganic acid.
15 Rep.~se.llalive salts and esters include the following:
Acetate, Ben7~n~s~1fonate, Benzoate, Bicarbonate, Bisulfate, Bitartrate, Borate, Bromide, Calcium Edetate, Camsylate, Carbonate, Chloride, Clavulanate, Citrate, Dihydrochloride, Edetate, Edisylate, Estolate, Esylate, Fumarate, Gluceptate, Gluconate, 20 Glutamate, Glycollylarsanilate, Hexyl~so~ a~e, Hydrabamine, Hydrobromide, Hydrochloride, Hydroxynaphthoate, lodide, Isothionate, Lactate, Lactobionate, Laurate, Malate, Maleate ,Mandelate, Mesylate, Methylbromide, Methylnitrate, Methylsulfate, Mucate, Napsylate, Nitrate, N-methylgl~ min~ ammonium salt, Oleate, Oxalate, Pamoate 25 (Embonate), Palmitate, Pantothenate, Phosphate/diphosphate, Polygalacturonate, Salicylate, Stearate, Sulfate, Subacetate, Succinate, Tannate, Tartrate, Teoclate ,Tosylate, Triethiodide and Valerate.
The term "pharmacologically effective amount" shall mean that amount of a drug or pharmaceutical agent that will elicit the 30 biological or medical response of a tissue, system, animal or human that is being sought by a researcher or clinician.
The term "alkyl" shall mean straight or branched chain alkanes of one to ten total carbon atoms, or any number within this range.

2~ 7465~ --- ~4 -The term "lower alkyl" shall mean straight or branched chain alkanes of orle to five total carbon atoms, or any number within this range. ~
The term "alkenyl" shall mean straight or branched chain alkenes with one o'r more degrees of unsaturation at any position on the chain, of two to ten total carbon atoms, or any number within this range.
The term "alkynyl" shall mean straight or branched chain alkynes with one or more degrees of u~lsi~lu~aLion at any position on the chain, of two to terl total carbon atoms, or any number within this range.
The term "aryl" shall mean phenyl, naphthyl or fluorenyl.
The term "cycloalkyl" shall mean cyclic rings of alkanes of three to eight total carbon atoms.
The term "trihaloalkylsulfonyloxo" shall mean the sllhstihl~nt o O
Whenever the term "alkyl" or "aryl" or either of their prefix roots appear in a name of a ~ ctihl~nt (e.g. aralkoxyaryloxy), it shall be interpreted as including those limitations given above for "alkyl" or "aryl." Designated numbers of carbon atoms (e g Cl-10) shall refer independently to the number of carbon atoms in an alkyl or cyclic alkyl moiety or to the alkyl portion of a larger substituent in which alkyl appears as it.s prefix root.
The terrn "oxo" shall refer to the substituent =0.
The term "halogen" shall include iodine, bromine, chlorine and fluorine.
The term "preterm labor" shall mean expulsion from the uterus of a viable il~fant before the normal end of gestation, or more particularly, onset of labor with effacement and dilation of the cervix before the 37th weel~ of gestation. It may or may not be associated with vaginal bleeding or rupture of the membranes.

-WO 95/14025 2 1 7 4 6 ~ ~ PC~/IJS94113483 The term "dysmenorrhea" shall mean painful menstruation.
The term "Caesarean delivery" shall mean incision through the abdominal and uterine walls for delivery of a fetus.
As used herein, the definition of each expression when it 5 occurs more than once in any structure, can be independent of its definition elsewhere in the same structure.
The term "substituted" shall be deemed to include multiple degrees of substitution by a named ~ub~ u~
Where multiple substituent moieties are disclosed or claimed, the sl-hstihlt~d compound can be independently sllhtitlltl~d by one or more of the disclosed or claimed substituent moieties, singly or plurally .
The compounds of the present invention, may have asymmetric centers and occur as racemates, racemic mixtures and as 15 individual diastereomers, or enantiomers with all isomeric forms being included in the present invention. Therefore, where a compound is chiral, the separate enantiomers, sllhst~nti~lly free of the other, are included within the scope of the invention; further included are all mixtures of the two enantiomers. Also included within the scope of the 20 invention are polymorphs and hydrates of the compounds of the instant invention.
Oxytocin (OT) and arginine vasopressin (AVP) are structurally related peptide hormones of the pituitary gland having distinct biological functions. OT is released from the pituitary in 25 response to stimuli related to parturition (e.g., labor, suckling of the neonate). Circulating OT then stimulates uterine activity to promote labor and delivery and contracts mammary gland myoepithelium to elicit milk-letdown postpartum. AVP, on the other hand, is secreted into the bloodstream by di~lu~ es in hemostasis such as reduced 30 blood pressure or blood volume and/or increased plasma osmolality.
AVP acts to correct these imh~ n(~ec by enhancing peripheral va.scular resistance and by promoting water reabsorption by the kidney. These effects of AVP are mediated by distinct receptors of the vascular smooth muscle (AVP-Vla receptor~) and kidney (Avp-v2 receptors), WO 9~i/14025 PCT/US94/13483 2 ~

respectively. AVP Vla receptors are also located in uterine smooth muscle to cause contraction and on platelets to mediate aggregation. By contrast, the contra~tile responses of the uterus and mammary glands to OT appear to be transduced by a separate, single receptor subtype. A
5 third, relatively obscure vas~ ,OOill receptor subtype, AVP-Vlb, has been identified in tlle pituitary gland where it mediates the stimulatory effects of AVP on adrenocorticotrophic hormone (AC~I) release. The human OT, Avp-vla and Avp-v2 receptors have recently been cloned and expressed.
The ability of the compounds of the present invention to antagonize oxytoc~ makes these compounds useful as pharmacologic agents for m~mm~l~, especially for humans, for the treatment and prevention of disorders wherein oxytocin may be involved. Examples of such disorders irnclude preterm labor and dysmenorrhea. These 15 compounds may also find usefulness for stoppage of labor ~l~paldtOIy to Cesarean deliver~.
The oxytocin antagonist compounds of the present invention are also Lseful for improving reproductive efficiency in farm animals. In certain farm animals (e.g., sheep, cattle, swine and goats), 20 the beginning of the estrous cycle is typically marked by behavioral estrus when the female animal accepts the male for mating. Ovulation of the ovarian follicle occurs shortly after onset of estrus and cells in the follicle give rise to the corpus luteum. The cells that form the corpus luteum produce progesterone and they also produce oxytocin.
25 The secretion of oxytocin from the corpus luteum and/or pituitary acts on the uterine endollletliulll to stimulate the secretion of pro.~t~ n-1inc (in particular PGF) which, in turn, causes the regression of the corpus luteum of the ovary. PGF is, therefore, the luteolytic hormone. In the cycling animal (i.e., where mating and fertilization have not occurred), 30 destruction of the corpus luteum removes the source of progesterone which is key to the preparation of the uterus for pregnancy. The presence of a viable conceptus (i.e., the embryo and its associated membranes) is necessary to prevent the luteolytic process. In fact, the first key signal that the conceptus must produce is the one to prevent WO95/14025 2 l 746~ PCT/US94113483 regression of the corpus luteum (i.e., the maternal recognition of pregnancy signal). Thus, in the animal where mating and fertilization have occurred, the conceptus secretes a factor that antagonizes the action of oxytocm to induce luteolysis. This results in ~ rl~AIloe of a S functioning corpus luteum and the continued secretion of progesterone which is obligatory to the initiation of pregnancy.
Adl.lilli~LIdLion of an oxytocin antagonist of the present invention at this critical period after fertilization (i.e., just prior to or during the period of maternal recognition of pregnancy) supplements o the natural signal from the conceptus (i.e., maternal recognition of pregnancy) to prolong corpus luteal function. The result is to increase pregnancy rates by enhancing the chances of impregnation through a reduction in embryonic loss. Thus, to improve fertility in a farm animal, a mated animal, for example, a mated ewe, IS treated with an oxytocin antagonist compound beginning on between day 10 to day 15 after onset of estrus. The oxytocin antagonist compound is ad~ L~Icd to the mated animal for a period of one day to three weeks, preferably one week to three weeks, most preferably one week to two weeks.
The compounds of the present invention are also useful in farm animals for controlling the timing of parturition so that delivery of neonates occurs during the daytime. Approximately 80% of livestock are delivered at night and up to 5 to 10% of newborns die because the deliveries are not monitored properly. An oxytocin antagonist compound of the present invention A~ lt;d to the mother on the evening before expected delivery delays parblrition so that the delivery occurs during the daylight hours. By delaying the timing of parturition, proper monitoring of the delivery and the neonates is ensured, resulting in increased survival rates of the newborns.
ln addition, the oxytocin antagonists of the instant invention can also be used to control the timing of estrus in a cycling farm animal by preventing luteal regression. An oxytocin antagonist compound of the instant invention is administered to a cycling farrn animal prior to expected e~strus to prevent regression of the corpus luteum. Daily = = = ~ , , WO 95114025 PCT/USg4/13483 21 74~5~ ~

administration of tlle compound retards estrus until administration of the compound ceases. Preferably, the oxytocin antagonist compound is t~ l,;d at lea~t I day prior to expected estrus. By delaying estrus in a group of farm animals, a farmer can synchronize estrus among the 5 group to provide time and cost savings in farm management.
The compounds of the present invention also bind to the vasopressin receptors and are therefore useful as vasopressin antagonists. Vasopressin antagonists are useful in the treatment or prevention of disease states involving vas~ disorders, including o their use as diuretics and their use in congestive heart failure.
It has llow been found that compounds of the present invention are balanced oxytocin and vasopressin antagonists useful for treating preterm labor and dysmenorrhea. The terms "balanced oxytocin and vasopressin antagonist(s)," "balanced oxytocin/vasopressin 15 compound(s)," and "balanced oxytocin/arginme-vasopressin (AVP)-VIa compound(s)," as used herein, are defined as a compound which has a ten-fold or less sep;lration between the binding affinity of the compound for a human oxytocin receptor and the binding affinity of the compound for a human AVP-`Vla receptor. Preferably, there is a five-fold or less 20 separation between the binding affinities of the compound to the oxytocin and AVP-Vla receptors. It is believed that a balanced oxytocin/vasopressin compound will provide a more effective treatment for preterm labor a~d dysmenorrhea. The fact that oxytocin receptors are up-regulated in the uterine myometrium during labor and before 25 and during menstruation (AVP-Vla receptors are also present but are not up-regulated), t~he observation that the uterus responds to both oxytocin and AVP and that the sensitivity of the uterus to the contractile effects of oxytocin is highest during labor and menstruation, and the fact that oxytocin/A.VP-stim~ t- d synthesis of contractile pros~ n-lin~;
30 occurs during labor and ~ lu~ion indicates that a balanced oxytocin/vasopressin antagonist compound would provide effective treatment for both preterm labor and dysmenorrhea. In addition, it appears that the vasoconstrictor activity of AVP (AVP-Vla response) contributes to uterine ischemia and pain. Thus, balanced WO95/14025 2 ~ 7 ~ PCT/US94/13483 oxytocin/v~sul,lc~ compounds, such as the balanced oxytocin/vasopressin compounds disclosed herein, provide a novel approach fûr the treatment of preterm labor and dysmenorrhea.
The compounds of the present invention can be 5 ~11minist~red in such oral dosage forms as tablets, capsules (each including timed release and sustdined release formulations), pills, powders, granules, elixers, tinctures, suspensions, syrups and emulsions.
Likewise, they may also be ~mini~t~red in imtravenous (both bolus and irlfusion), intraperitoneal, subcutaneous or intramuscular form, all using forms well known to those of ordinary skill in the pharmaceutical arts.
An effective but non-toxic amount of the compound desired can be employed as a tocolytic agent.
The dosage regimen utili~ing the compoumds of the present invention is selected in accordance with a variety of factors including 15 type, species, age, weight, sex and medical condition of the patient; the severity of the condition to be treated; the route of administration; the renal and hepatic function of the patient; and the particular compound or salt thereof employed. An ordinarily skilled physician or v~le~illdli~l can readily d~L~ li"e and prescribe the effective amount of 20 the drug required to prevent, counter or arrest the progress of the condition.
Oral dosages of the present invention, when used for the indicated effects, will range between about 0.3-6.0 gm/day orally.
Intravenously, the most preferred doses will range from 0.1 to about 10 25 mg/minute during a constant rate infusion. Advantdgeously, compounds of the present invention may be administered in a single daily dose, or the total daily dosage may be administered in divided doses of two, three or four times daily. Furthermore, preferred compounds for the present invention can be administered in intranasal forrn via topical use 30 of suitable intranasal vehicles, or via transdermal routes, using those forms of transdermal skin patches well known to those of ordinary skill in that art. To be ~ll",illi.~ d in the form of a transdermal delivery system, the dosage administration will, of course, be continuous rather than intermittant throughout the dosage regimen.
. . _ ~

WO 9~/14025 PCrNS94/13483 ~ 14~
- 3() -In the ]nethods of the present invention, the compounds herein described in detail can forrn the active ingredient, and are typically ~rlminist~led in admixture with suitable pharmaceutical diluents, excipients or carriers (collectively referred to herein as 5 "carrier" materials) suitably selected with respect to the intended form of administration, tllat is, oral tablets, capsules, elixirs, syrups and the like, and consistl~nt with conventional pharm~e-lticAI practices.
For instance, for oral adl.lil.i~L-~llion in the form of a tablet or capsule, the acti~e drug component can be combined with an oral, non-toxic pharm~el-tir~lly acceptable inert carrier such as ethanol, glycerol, water and the like. Moreover, when desired or necessary, suitable binders, lubricants, disintegrating agents and coloring agents can also be incorporated into the mixture. Suitable binders include starch, gelatin, natural sugars such as glucose or beta-lactose, corn 5 sweeteners, natural and synthetic gums such as acacia, tragacanth or sodium alginate, carboxymethylcellulose, polyethylene glycol, waxes and the like. Lubricants used in these dosage forms include sodium oleate, sodium stearate, m~n( sillm stearate, sodium benzoate, sodium acetate, sodium chloride and the like. Disintegrators include, without 20 limit:~tinn, starch, methyl cellulose, agar, bentonite, zanthan gum and the like.
The compounds of the present invention can also be adl..i--i~ d in the IForm of liposome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles and m~ m~llar 25 vesicles. Liposome3 can be formed from a variety of phospholipids, such as cholesterol, stearylamine or phosphatidylcholines.
Compounds of the present invention may also be delivered by the use of monoclonal antibodies as individual carriers to which tlle compound molecules are coupled. The compounds of the present 3~ invention may al.so be coupled with soluble polymers as targetable drug carriers. Such polymers can include polyvinylpyrrolidone, pyran copolymer~ polyhydroxypropylmethacrylamidephenol, polyhydroxyethylasl~artamidephenol, or polyethyleneoxidepolylysine substituted with palmitoyl residue.~. Furthermore, the compounds of the WO 95/~4025 2 1 7 4 6 ~ G PC170S94/13483 present invention may be coupled to a class of biodegradable polymers useful in achieving controlled release of a drug, for example, polylactic acid, polepsilon caprolactone, polyhydroxy butyric acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates and cross-linked or a llL,llil,alllic block copolymers of hydrogels.
The present invention is also directed to combinations of the compounds of formula I with one or more agents useful in the treatment of oxytocin related disorders such as preterm labor, dysmenorrhea and stopping labor prior to cesarean delivery. For example, the compounds of the instant invention may be effectively administered in combination with effective amounts of other agents used in the treatment of preterm labor, such as antenatal steroids (e.g., ~irxi1~,rlllAcolle). Preferred combinations are ~imlllt~n~ous or alternating tlca~lllC.Il~, of an oxytocin receptor antagonist of the present invention and an antenatal steroid. These combinations have beneficial effects on the neonate by both decreasing uterine activity to prolong gestation and increasimg fetal maturation. In accordance with the method of the present invention, the individual components of the combination can be A(lmini~tered separately at different times during the course of therapy or concurrently in divided or single combimation forms. The instant invention is therefore to be understood as embracing all such regimes of ~imllltAn~ous or alternating treatment and the term "A~lminictrring" is to be interpreted accordingly. It will be understood that the scope of combinations of the compounds of this invention with other agents useful for treating oxytocin related conditions includes in principle any combination with any ph~rmArelltirAl composition useful for treating preterm labor, dysmenorrhea or stopping labor prior to cesarean delivery.
The compounds of the instant invention can be prepared readily accordin~ to the following reaction schemes and Examples or modifications the,-eof using readily available starting materials, reagents and conventional synthesis procedures. In these reactions, it is also possible to make use of variants which are themselves known to those of ... . . .. .. .. _ _ _ _ _ _ WO 95114025 PCrNS94/13483 2~ 74~

ordinary skill in th] s art, but are not mentioned in greater detail.
Reference can also be made to U.S. Patent No. 5,091,387, issued February 25, 1992, the entire disclosure of which is incorporated by reference. Additiollal reference can also be made to the method of Matier, et al., J. Org. Chem., Vol. 36, No. 5, 650-654 (1971), the entire disclosure of which is incorporated by reference, on elaboration of indenes and their 2-oxo derivatives to ~,uilului~elidine analogs. Also irlcorporated by reference is a variant of this procedure described in Matier, et al., J. Org. Chem., Vol. 36, No. 5, 650-654 (1971).
0 The most preferred compounds of the invention are any or all of those specifically set forth in these Examples. These compounds are not, however, ta be construed as forming the only genus that is considered as the invention, and any combination of the compounds or their moieties may itself form a genus. The following examples further 5 illustrate details for the preparation of the compounds of the present invention. Those sk.illed in the art will readily understand that known variations of the cnrl~iitic)nc and processes of the following preparative procedures can be used to prepare these compounds. All ~tillll)eldLUl~S
are degrees Celsius lmless noted otherwise.

3~
-WO 951140~5 2 1 7 4 ~ 5 0 Pcr/uss4ll34s3 Abbreviations used in the Examples are as follows:
BOP = bell~,u~lid/ul-l-yloxytris(dimethylamino)phosphonium hexafluorophosphate 5 DCM = dichloromethane DIEA = diisu~-u~lethylamine DMF = dimethylformamide EtOAc = ethyl acetate EtOH = ethanol EDC = l-ethyl-3-(3-dimethylaminopropyl)carbodiimide FAB MS = fast atom bombardment mass spectroscopy HOBT = 1-hydroxybenzotriazole HPLC = high pressure liquid chromatography MeOH = methanol 15 NMR = nuclear magnetic resonance THF= tetrahydrofuran TLC = thin layer chromatography 21 7465~

lEXAMPLE 1 `~O2C~

clcocH2CH=cH2 ~5O2C~
H NHCOCH2CH=CH2 WO 9511402~ PCTIUS94/13483 21 7~5~

5 ~
SO2C~

ClSO2N(CH3)3 H N
so2CH2 H NHSO2N(CH3)2 ~46~ --~O2C' ~

ClCO2CH2CH3 20 5O2CH~
2s H NHCO2CH2CH3 W095/14025 2 1 7 4 ~ 5 ~

~02C~

CICO~

:~o C~ ~
so2CH2 NHC
H

wo 95/~402s 2 1 ~

~O~C~

CICO~
N

2 o 52C~
H NHCO~N

21 74S~i~

~02C~/

CO2H ,~
Step 1 Step 2 2s ~ ~ A, SO2CH2 /~
H NHCO~ ¦

PC~IU594/13483 wo 95/141)25 2~ 7~

HO2CC H2--N ,N

2~ ~ `r' SO2CH2 HCOCH2--N' ,N
\=N

, I--II~Y 217465C PCT~US 94/13483 ~1 Rec'd PCT/PTO 2 4 ~T~995 REACTIONS~HFMF`S PART2 ~02C~

CICO

`~O2C~
H NHCO/~

ll,lFND~D S~

rr~Y
21 74~5G ~ q 4~r 1 7 ~
~1 Re~'d ~CTIPTO 24 OCT19~5 ~02CH~

HNOC"
H~
H o So~CH~$
H 'N- C

wo 1 21 74650 REACTION SCHEMES. PART 3 .o2cH2~

N ~
so2CH2 I R~ Y 2 1 7 ~GT a ~ I 1 3 ~ 8 2 4 aGT

REACTION SCHEMES, PART 3 cont'd SO2CH~

HO~O
to ~N~

SO2C~ ,,~OCH
H NHCO Nl co2c(CH3)3 S02C~> ~c~OH
H NHCO N
C02C(CH 3)3 ~MENDED SllEEr ,~Y 21 74~PCT~US 9~/13483 " ~ .' 2~ GCT1995 REACTIQN S~HFMF~ PART 3 (~ONTD

~O2C~
H NHCO N
co2c(CH3)3 HCI

~' S02CH 2 , H NHCO N
H HCI
~, ;,F~

WO 9~/1402!; PCT/US94113483 21 7465'J

REACTION SCHEMES. PART 3 CONT'D
H N
so2CH2 H

so2CH2 H NHCOCH
N

H

~VO 9~11402~i PCT/US94/13483 2 ~ 7 ~b~O

5 ~ ~
so2CH2 H NH
o 2 N

H NHCOCH2~
0 NHCO2C(CH3)3 W11)95/14025 2 ~ l'CT/US94/13483 .

REACTION SCHEMES. PART 4 CONT'D

5 ~`J~

H NHCOCH2~
o NH2 HCI

21 7465~ --REACTION SCHEMES~ PART ~
@~ ~ HOOCCHCH2~NH
~ NHCO2C(CH3)3 so2C~I2 y, (, N=\
H NHCOCHCH2~,NH
NHCO2C(cH3)3 ~2C~, N
H NHCO IHCH2~,NH

WO g5/14025 PCI'IUS94/13483 2~ 74~150 REACTION SCHE~F.`~. PART 5 CONT'D
~, NHCO2C(CH3)3 '~ HOOClHCH2~qCH

~,~
SO2CH2 HCOfHCH2~
NHCO2C(CH3)3 2s EXAMPLE 19 H ~HcocHcH

=-- ~
WO 95/14025 PCT/~IS94~13483 2 1 ~ U

]~EACTION SCHl~MF~. PART 6 [~ HOOCCH2--N~) ~ ~ NHC02C(CH3)3 s02CH2 ,y"

SO2CH2 y.

~--<
NHco2c(cH3)3 H NHCOCH2--N~

WO 9~/14025 2 1 7 4 6 5 ~ PCT/US94/13483 REACTION SCHEMES. PART 7 (CICH2CH2)2NH HCI + [(CH3)3COC0]20 Et3N
(ClCH2CH2)2NCO2-t-Bu ~ + (ClCH2CH2)2NCO2-t-Bu LiN[Si(CH3)312 N~o t Bu HCI, EtOAc WO 95/14û25 PCT/US94/13483 ~ 1746~

~F~CTION SCHEMES. PART 7 CONT'D
5 [~ + r~
H HCI O

Et3N

52 ~
O

-WC 95114025 ~ 1 74 ~ PCTIUS94113483 - ~3 -REACTION SCHEMES. PART 7 CONT'D

pyridine N-OH
Ni(R), H2 ' 3 i~

3c Endo Isomer WO 95/14~25 PCT/US94/13483 - ~4 -RE~ACTION SCHEMES. PART 7 CONTD

H
EDC, HBT, NEt3, DMF
lo 2.
HCI
~2 ~H
HN~ N~, O N~
H HCI

III~Y 217465~ PCT/US 94/1~483 51 Re~'d P~P~O 24 ~rl9~S

REACTION SC~ . PART X
H HCI
CISO\.
N Et3 N
SO2~
Benzene AMENDED SltE~T

~7~
- ~6 -RF.~CTION SCHEMES. PART ~ CONT'D
N H~

+ TRACE OF EXO ISOMER
1. BH3 THF
H202, OH
2. pyridinium chlorochromate, ~V095/14025 21 74650 PCT/US94/13483 REACTION SCHEMES. PART 8 CONrD
- 5 ~J~

Anthracene Toluene I2 - L_ ~r ~

WO 9~/14025 PCT/US94/13483 2 1 ~ 4 ~
~F~CTION SCHEMES~ PART 8 CONT'D
'~ 1~>
so2,o ~ Q
~INI~ L~

_ 59 _ REACTION SCHEMES. PART ~ CONT'D
so2,~

2 s FXAMPLE 34 ' ,A ~

.

p ~ ~ 2 4 ~G ~ ,~S

REA(3TIONSCHFI~F~ PART9 N
SO2~
V--OH
Toluene [~ ~f ~ CH3 -->

~MENDEI) SHEET

REACTION SCHEMES~ PART 10 1. Methyliminodiacetic acid anhydride, toluene THF
2. CH31, DMF

7\~oH

~0 2 5 <
c02CH3 WO 9~/14025 -- PCT/US94113483 2~74650 RE~CTIQN SCHEMES. PART 11 Nl 2 ~ ~o;~~

~CT/US 94/13483 61 R~c'd PCTIPTO 24 OCT199S

RF~CT~ON ScHFl\/lF~ PART 11 CQN~D
¢~
N

, \
Zn - HOAc N
E~AMpl F 3 ~MENDED ~H~

Wo 95114025 PCI~/US94/13483 21 7~650 EXAMPLE A
~02CH2~
o '"'NH
Endo-(1 S)- I '(((2-amino-7,7-dimethylbicyclo(2.2. 1 )-hept- I -yl)-melllyl)-sulfonyl)~piro(lH-indan-l .4'-piperidine) Di-t-butyl dicarbonate (31g, 0.14 mole available from Aldrich) and bis(2-chloroethyl)amine hydrochloride (21.6g, 0.12 mole 5 Aldrich) were combined in CH2C12 (250 ml) stirred at ambient ldLul~ and tr~ated with triethylamine (12.8 g, 0.127 mole) added dropwise over 15 minutes. After I hour, another 1.5 ml of triethylamine was added. After a total of 2.5 hours, the mixture was poured onto a silica gel column packed with CH2C12:hexane 20 (1:1), and eluted ~ith CH2C12. The combined product fractions were evaporated to dryness in vacuo to give N,N-bis(2-chloroethyl)-t-butyl-carbamate.
To a solution of indene (10.3 g, 89 mmole) in dry tetrahydrofuran (TMF, 18 ml) cooled in an ice bath and maintained 25 under a nitrogen blanket was added lithium bis(trimethylsilyl)amide (Aldrich, 177 ml of a l.OM ~olution in THF; 177 mmole) over 15 minutes. The mixl:Ure was stirred in the cold for 30 minutes, then added over 15 minutes to a solution of N,N-bis(2-chloroethyl)-t-butylcarbamate (2~1.2 g, 88 mmole) stirred in an ice bath. The mixture 30 was ~tirred for 2 h~urs in the cold and for 30 minutes at ambient temperature under nitrogen, then evaporated in vacuo to a foam.
CH2C12 was added and the resulting mixture poured onto a silica gel column packed with 40% hexane in CH2C12. The column ~, ?,, ~

~0 9S/1402~i PCT/~JS94/13483 was eluted with 40~o hexane m CH2cl2 followed by CH2C12, and the product fractions were evaporated to dryness in vacuo to provide l'-(t-butyloxycarbonyl)-spiro(indene- 1 ,4'-piperidine).
- 1'-(t-Butyloxycarbonyl)spiro(indene-1,4'-piperidine) (16 g, 5 56 mmole) in ethyl acetate (250 ml) was stirred in an ice bath and saturated with HCl(g) for 30 minutes. The mixture was evaporated to dryness. Ethyl acetate was added and removed in vacuo three times, and the residue was triturated with diethyl ether and filtered to provide spiro(lH-indene-1,4'-piperidine) hydrochloride. The free base was obtained by slurrying the hydrochloride in aqueous sodium bicarbonate solution and extracting with CH2cl2. The organic layer was separated, dried over sodium sulfate, filtered, and evaporated to dryness in vacuo to provide spiro(lH-indene-1,4'piperidine.
Spiro(lH-indene-1,4'piperidine) (308 mg, 1.66 mmol) and 15 (+)-lo-~a~ hol~ulfonyl chloride (418 mg, 1.66 mmol) were combined in CH2C12 and treated with triethylamine (0.23 ml). The mixture was stirred at ambient ~ ldlUI~ for 15 minutes, then poured onto a silica gel column and eluted with 1:1 CH2cl2:hexane. The product fractions were combined and evaporated to dryness in vacuo to provide (IS)-I'-2 (((7,7-dimethyl-2-oxobicicylo-(2.2. 1 ) hept- I -yl)-methyl)sulfonyl)spiro-(lH-indene-1.4'-piperidine) as a solid which was recrystallized from petroleum ether and dried ovemight in vacuo at ambient l~ ,elalul~.
( 1 S)- 1 '-(((7,7-dimethyl-2-oxobicyclo(2.2. 1 )hept- 1-yl)methyl)sulfonyl)spiro(lH-indene-1,4'-piperidine) (30 g, 0.075 mole) 25 irl pyridine (500 mL) was heated in an oil bath to 70C (internal).
Hydroxylamine hydrochloride (30 g) was added in three portions over ca. 20 minutes. After 2 hours, an additional 10 g of hydroxylamine hydrochloride was added (over 10 minutes). At 30, 40, and 50 minutes additional elap~ed time, further 3 g lots of hydroxyl-amine 30 hydrochloride were added. After another 30 minutes, the mixture was poured into water (2 L) and extracted 3 times with ethyl acetate (300 mL portions). The organic layers were combined, washed with I N HCI
(600 mL total), dried over sodium sulfate, filtered, and evaporated to dryness in vacuo. EtOH (abs; ca. 250 mL) was added to the resulting WO 95/14025 PCrJUS94113483 .
~7 thick syrup and the solution allowed to stand at ambient temperature overnight. The m~xture was filtered aLLd the fLltrate boiled down to ca.
80 mL. After standing, the mixture was again filtered and boiled down to ca. 20 mL. After a third filtration, the filtered solids were combined to give (lS)-1'-(((7,7-dimethyl-2-oximinobicyclo(2.2.1)hept-1-yl)-methyl) sulfonyl)spiro(lH-indene-1,4'-piperidine) (28 g).
Fresh~y prepared, activated Raney Nickel catalyst (ca. 30 g) in water was allowed to settle and the water decanted. Abs. ethalLol (300 mL) was added, and the mixture swirled and again allowed to settle. The solven~ was decanted. lwo more wash-decant cycles with 150 mL of ethanol were similarly carried out. (lS)-1'-(((7,7-dimethyl-2-oximinobicyclo(2.2.1)hept-1-yl)methyl)sulfonyl)-spiro(lH-indene-1,4'-piperidine) (30 g) was stirred in a mixture of abs. ethanol (450 mL) and 2-methoxyethanol (900 mL), nitrogen was bubbled through the 5 suspension/solutioll, and the Raney Nickel catalyst was added. The mixture was hydrogenated under 50 psi overlLight. TLC (9:1 CH2C12MeOH, silica gel) showed the reaction to be complete. The catalyst was remo~led by filtration, and the filtrate evaporated to dryness in ~Ç~Q. The crlLde solid (27 g) was divided into 7 g batches, and each 20 batch was dissolved in methylene chloride (ca. 200 mL) and flash c~Lromatographed on silica (700 g in a 100 mm column, packed and eluted with 8% (v~v) methanol ilL methylene chloride), taking 200 mL
fractions. The exo isomer of the title amine was obtained in fractions ca. 5-7, and the desired endo isomer in fractions ca. 8-16. TLC was on 25 silica, eluted with 8% methanol-methylene chloride, phosphomolybdic acid stain. The combined product fractions were evaporated to dryness to provide the title compound (4.5 g from each 7 g lot, ca. 18 g total) a.s a colorless solid.

r WO 95/14025 PCI/~JS94/13483 21 7465~

EXAMPLE I

N
s02CH2 \~ H
0 HN~
o Endo-(lS)-1'-(((2-(but-3ene-1-ylamino)7,7-dimethyl-bicyclo(2.2.1)-hept-l -vl)-methyl)sulfonyl)~piro(lH-indan-1 .4'-piperidine) To a solution of endo-lS-1'-(((2-amino-7,7-dimethyl-bicyclo(2.2. 1 .)-hept- I -yl)-methyl)-sulfonyl)spiro(l H-indan- 1,4'-piperidine) (50 mg, 0.000125 m) in methylene chloride (3 ml) in an atmosphere of nitrogen was added crotonyl chloride (13.1 mg, 14 ml, 0.000125 M) via a syringe. After two minutes, triethylamine (50 ml) 20 was added to make the solution basic (ph=9). After I hour, the reaction mixture was concentrated and the oil was chromatographed on a silica "flash" column using 40% ethyl acetate in hexane as solvent. Fraction~
27-40 contained a fluorescent spot. These were concentrated and ether-hexane was added and collc~ aled to yield the title compound 25 as a white foam.
NMR (CDC13) was c~llsi~ lt with structure.
HPLC~ 210-97 .181% 254-99.3 86%
Mass Spectra: Calculated, m/e-470.679; Found, m/e=471.3 30 Analysis c~ t~d for C27H3gN2O3S
C, 68.90; H, 8.14; N, 5.95 Found: C, 68.99; H, 8.19; N, 5.7 wo 95/14025 PCr~sg4/l3483 2~ 7465~

SO ~, .

- I
HN `SO' N `CH
Endo-( 1 S)- I '-(((2-(dimethylaminosulfonylamino)-7 ,7-dimethyl-bicyclo(2.2. 1 )-hept- I -yl)-methyl)-sulfonyl)spiro( I H-indan- 1,4-idine) In an oven-dried flasl; (50 ml) under nitrogen was dissolved endo- I S- I '-(((2-amino-7,7-dimethylbicyclo(2.2. 1 .)-hept- I -yl)-methyl)-sulfonyl)-spiro(lH-indan-1,4'-piperidine)(50 mg, 0.000125 M) in methylene chlor;de (3 ml). Dimethylaminosulfonyl chloride (17.9 mg, 0.000125 M) was added, and after two minutes, triethylamine (50 ml) was added. After one hour, the reaction mixture was concentrated under reduced pressure (20 mm). A silica "flash" column using methylene chloride(9)-methanol(l) as solvents was used to purify the title compound as a tan solid after adding and concentrating with hexane.
NMR (CDC13) was con~ l with structure.
HPLC: 210-96.4Z% 254-100%
Mass Spectra: Calculated, 509.736; Found, 510.3 Analysis calculated for C2sH3gN3O4S2 C, 5951; H, 7.93; N, 8.04 Found: C, 59.58; H, 7.71; N, 7.87 wo 9~/14025 PCrN594113483 21 74~5~

SO ~.
HN\I~O~/
Endo-( 1 S)- I '-(((2-(ethoxycarbonylamino)-7,7-dimethyl-bicyclo-(2.2.1 )-hept- I -vl)-methYI)-sulfon~d)spiro( 1 H-indan- 1 .4-piperidine) ~ an oven-dried flask (50 ml) under nitrogen was dissolved endo- 1 S- I '-(((2-amino-7 ,7-dimethylbicyclo(2.2. 1 .)-hept- I -yl)-methyl)-sulfonyl)-spiro(lH-indan-1,4'-piperidine)(S0 mg, 0.000125 M) in methylene chloride (3 ml). Ethyl chloluru~ a~ (13.6 mg, 12 ml, 0.000125 M) was âdded via syringe. After two minutes, triethylamirle (50 ml) was added to make the solution basic (pH=9). After 30 minutes, the reaction mixture was concentrated under reduced pressure (20 mm).
The oil was chromatographed on a silica "flash" column with 40% ethyl acetate in hexane as eluent. Fractions 12-20 contained the product.
These fractions were concentrated under reduced pressure (20 mm).
Hexane-ether was added to the oil and removed under reduced pressure to yield 17 mg of the title compound as a white powder.
NMR (CDC13) was consi~ with structure.
HPLC 210-99.~17% 254-100%
Mass Spectra: Calculated 474.668, Found 475.3 Analysis c:~lrlll~t~d for C26H3gN2O4S
C, 65.79; H, 8.07; N, 5.90 Found: C, 6G.05; H, ~.22; N, 5.72 WO 9~/14025 PCrllJS94/13483 ~1 74~5~

EX~MPLE 4 ~`32-~
o H~ ~
Endo-(lS)-I '-(((2-~2-thiophenecarbonylamino)-7,7-dimethylbicyclo-(2.2.1)-hept-1 -vl)-~nethyl)-sulfonyl)spiro(lH-indan-l .4-piperidine) In an oven-dried flask (50 ml) under nitrogen was dissolved endo- I S - I '-(((2-amino-7,7-dimethylbicyclo(2.2. 1 .)-hept- I -yl)-methyl)-sulfonyl)-spiro(lH-indan-1,4'-piperidine)(50 mg, 0.000125 m) in methylene chloride (3 ml). The 2-thiophene carbonyl chloride (18 20 mg, 13.4 ml, 0.000125 m) was added via syringe. Triethylamine (50 ml) was added to ni~ake the mixture basic. The reaction mixture was concentrated to oil and chromatographed using a silica "flash" column with 40% ethyl acetate in hexane as eluent. Fractions 12-20 contained the product. Fractions 11-18 contained the desired product and these were corl~cntr~t~d to an oil. Ether-hexane was added and removed to yield 27 mg of the title compound as a white amorphous powder.
HPLC 210-98.873% 254-97/931%
Mass Spectra: Cal~ulated, 512.738; Found, 513.3 Analysis calculated for C28H36N203S2 0.72 hexane C,63.~7; H,7-11; N,5-33 Found: C, 63~9; H, 6.7~; N, 5.08 W~ 95/14025 PCINS94/13483 21 7465a ~l2 HN~
Endo-(lS)-I '-(((2-(isonicotinylamino)-7,7-dimethyl-bicyclo-(2.2.1 )-hept- 1 -yl)-methyl)-sulfonyl)spiro( I H-indan- 1 ~4-piperidine) In an oven-dried flask (50 ml) under nitrogen was dissolved endo- 1 S- I '-(((2-amino-7,7-dimethylbicyclo(2.2. 1 .)-hept- I -yl)-methyl)-sulfonyl)-spiro( l H-indan- 1 ,4'-piperidine)(50 mg, 0.000 125 m) in methylene chloride (3 ml). The isonicotinoyl chloride dihydro-chloride (22.38 mg, 0.000125 m) was added. After 2 minutes, triethylamine (50 ml) was added so the pH equaled 9. The solvent was removed under reduced pressure and the product was purified by a "flash" silica column using methylene chloride (9)-methanol (1) as solvents. The product was contained in fractions 6-10 which were cnnl çntr~t~d. The oil was treated with hexane-ether which was removed under reduced pressure. This gave 18 mg of the title compound as a white foam.
NMR (CDC13) was CollSi~ with structure.
HPLC 210-95.84% 254-98.04%
3 Mass Spectra~ lc~ tPrl 507.7; Found, 508.3 Analysis calculated for C2gH37N3O3S
C, 68.61; H, 7.35; N, 8.28 Found: C, 6~.46; H, 6.99; N, 7.90 WO 9~14025 PCT/US94/13483 2l 7~

~O~c5i~
lo HN
o Endo-( 1 S)- I '-(((2-(noradamantyl- 1 -carbonylamino)-7,7-dimethyl-bicyclo(2.2. 1 )-hept- I -yl)-methyl)-sulfonyl)spiro( I H-indan- 1,4-piperidine) Step 1: Noradamantyl-l-carbonyl chloridç
To a solution of noradarnantylcarboxylate (20.8 mg, 0.000125 M) in melhylene chloride (4 ml) in a round bottom flask (50 ml) under an atmos]?here of nitrogen at 0C was added oxalyl chloride (120 ml) via syringe. Dimethylformamide (7 ml) was added and the solution was stirrçd at 0C for 15 minutes. The reaction mixture was allowed to warm to room ~ ul~ and stir for 45 minutes. The solvent was removed under reduced pressure (20 mm). Twice ?"1~1iti~n:~1 methylene chloride was added and removed under reduced pressure. The acid chloride of the title compound was kept under high vacuum (0.05 mm) for one hour before using in the next step.
Step 2: Endo-( I S)- I '-(((2-(noradamantyl- 1 -carbonylamino)-7, 3 7-dime~hylbicyclo(2.2. 1 )-hept- I -yl)-methyl)sulfonyl)-~piro( I H-indan- I ~4'-piperidine) Adamalltyl-l-carbonyl chloride was dissolved in methylene chloride (4 ml) and endo-lS-1'-(((2- amino-7,7-dimethylbicyclo(2.2.1)-hept- I -yl)-methyl)-sulfonyl)spiro( I H-indan- I ,4'-piperidine) (50 mg, 0.000125 m) was added. After two minutes, triethylamine ( 100 ml) wa~s 2~ 7465G

introduced via syringe. The solvent was removed under reduced pressure and the oil was chromatographed on silica with 40% ethyl acetate in hexane as elutant. The product fractions were collected and concentrated to an oil, which became an amorphous white solid upon 5 addition and removal of ether-hexane. This gave 10 mg of the title compourld.
NMR (CDC13) was COII~ ell~ with structure.
HPLC 210-98.427% 254-100%
o Mass Spectra: Calculated, m/e 550.81; Found, m/e 551.3 Analysis calculated for C33H46N2O3S
C, 71.96; H, 8.42; N, 5.09 Found: C, 71.57; H, 8.71; N, 4.77 H NHJ~O
O
Exo-(lS)-1'-(((2-(ethyl succinoylamino)-7,7-dirnethylbicyclo(2.2.1)-hept-l -Yl)-methvl)-.sulfonyl)s~iro(lH-indan-l .4'-pireridine) To a solution of exo-(lS)-1'-(((2-(amino)-7,7-dimethylbicyclo(2.2.1)-hept-1-yl)methyl)-sulfonyl)spiro(lH-indan-1,4'-30 piperidine) (50 mg, 0.000125 M) in methylene chloride (3 ml) under ablanket of nitrogen was added ethyl succinoyl chloride (20.6 mg, 0.000125 M). After two minutes, triethylamine (50 ml) was added with a syringe. The pH of the solution was 9. After 15 minutes, a new spot wa~s observed on the tlc (silica~0% ethyl acetate in hexane). The reaction mixture was concentrated and the oil was chromatographed on WO 9!i/14025 PCIIUS94/13483 .
21 7~650 a silica "flash" column with 40% ethyl acetate as solvent. Fractions 24-48 contained the product. These fractions were collected and concentrated. Eth~r-hexane was added and removed to yield 21.3 mg of the title compound as a white powder.
NMR (CDC13) was consistent with structure.
HPLC 210-99.190% 254-96.914%
Mass Spectra: Calculated mle, 530.732; Found m/e, 531.5 Analysis calculated for C2gH42N2OsS 0.25 H2O
C, 65.08; H, 8.00; N, 5.23 Found: C, 65.16; H, 8.03; N, 5.09 '2C~ H N =N
HN ~ ~N
o Endo-( 1 S)- I '-(((2-(tetrazole- 1 -acetylamino)-7 ,7-dimethylbicyclo-(2.2.1)-hept-1-vl)-meth~ ulfonyl)-spiro(lH-indan-l.4-piperidine) In an oven-dried flask (50 ml) under nitrogen was dissolved endo-1$-1'-(((2-amino-7,7-dimethylbicyclo(2.2.1.)-hept-1-yl)-methyl)-sulfonyl)-spiro(lH-indan-1.4'-piperidine)(50 mg, 0.000125 M).
tetrazole-l-acetic acid (16 mg, 0.000125 M), and benzotriazol-l-yloxytris(dimethylamino)phosphonium-hexafluorophosphate (Sequalog) (55 mg, 0.000125 M) in acetonitrile (3 ml). Diisopropylethylamine (50 ml) was added to make the solution basic. After tlc (silica-methylene chloride(9)-methallol(l)) showed a new spot, the reaction was concentrated to an oil. The oil was dissolved in methylene chloride--Wl)95/14025 PCrlUS94113483 ether (1:3) and the solution was washed with water, sodium bicarbonate (sat., aqueous), water, 10% potassium bisulfate, and brine. After drying with sodium sulfate, the solution was filtered and concentrated to oil, which was chromatographed on a silica "flash" column with 10%
5 methanol in methylene chloride. Fractions 5-10 contained the product and were collected and concentrated. Ether-hexane was added and removed to yield the title compound as a white powder.
NMR (CDC13) was consistent with structure.
HPLC: 210-98% 254-100%
Mass Spectra: Calculated, 513.679; Found, 513.3 Analysis c~ ted for C26H36N6O3S 0-25 H2O
C, 60.37; H, 7.11; N, 16.25 Found: C, 60.28; H, 6.96; N, 16.18 EX~MPLE 9 20 ~ ~

HN~o o Endo-( 1 S)- I '-(((2-(2-furoylamino)-7 ,7-dimethylbicyclo-(2.2. 1)-hept- I -yl)-methyl)sulfonyl)spiro( I H-indan- I .4-piperidine) In an oven-dried flask (50 ml) under nitrogen was dissolved endo- 1 S- I '-(((2-arnino-7,7-dimethylbicyclo(2.2. 1 .)-hept- I -yl)-methyl)-sulfonyl)-spiro(lH-indan-1,4'-piperidine)(50 mg, 0.000125 M) in methylene chloride (3 ml) and 2-furoyl chloride (16.3 mg, 0.000125 M) was added. After two minutes, triethylamine (150 ml) was added so the pH equaled 9. The solvent wa.s removed under pressure and the ,, _ _ , . _ . , . _ . . . ... . . . . . ..

WO 95114025 PtCTlUS94/13483 .

product was purified by a silica "flash" column using 40% ethyl acetate in hexane as solvents. The product was contained in fractions 6-10 which were concentrated. The oil was treated with hexane-ether which was removed under reduced pressure.

NMR (CDC13) was consistent with structure.
HPLC: 210-99.341% 254-99.576%
Mass Spectra~ It~ (l, 496.674; Found, 497.3 Analysis calculated for C2gH36N2O4S
o C,67.71;H,7.31;N,5.64 Found: C, 67.~1; H, 7.39; N, 5.46 ~o2cH2 7 \ /, HN
o Endo-(lS)-1'-(((2-(S-(1)-2-oxo-pyrrolidin-5-yl-carbonyl)amino)-7,7-dimethylbicyclo(2.2.1)-hept-1-yl)-methyl)-sulfonyl)spiro(lH-indan-I ~4-piperidine) In an oven-dried flask (50 ml) under nitrogen was dissolved endo- I S- I '-(((2-amino-7,7-dimethylbicyclo(2.2. 1 .)-hept- I -yl)-methyl)-sulfonyl)-spiro(lH-indan-1,4'-piperidine)(50 mg, 0.000125 M), S-(1)-2-oxo-pyrrolidine-5-carboxylic acid (16.1 mg, O.OOOI'25 M), and benzotriazol- I -yloxytris(dimethyl-amino)phosphonium-hexafluoro-phosphate (Sequalo~) (55 mg, 0.000125 M) in acetonitrile (3 ml).
Diisopropyl-ethylamine (50 ml) was added to make the solution basic.
After tlc(silica-methylene chloride(9)-methanol(l)) showed a new SpOt.

WO 95114025 PCrlUS94113483 the reaction was concentrated to an oil. The oil was dissolved in methylene chloride-ether(1:3) and the solution was washed with water, sodium bicarbonate (sat., aqueous), water, 10% potassium bisulfate, and brine. After drying with sodium sulfate, the solution was filtered and 5 concentrated to an oil. Ether-hexane was added and removed under reduced pressure to give the title compound as a white solid.
NMR (CDC13) was consistent with structure.
HPLC: 210-93.102% 254-92.292%
0 Mass Spectra: Calculated, 513.705; Found, 514.3 EXAMPLE l I
\~
~0 o Endo-(lS)-I '-(((2-(L-N-(tert.-butoxycarbonyl)-4-benzyl-oxy-2s prolinoylamino)-7,7-dimethylbicyclo(2.2. 1 )-hept- I -yl)-methyl-.sulfonyl)~spiro( I H-indan- l ~4-piperidine) In an oven-dried flask (50 ml) under nitrogen was dissolved endo-lS-1'-(((2-amino-7,7-dimethylbicyclo(2.2.1.)-hept-1-yl)-methyl)-sulfonyl)spiro( I H-indan- I ,4'-piperidine)(50 mg, 0.000125 M), 30 L-N(tert.-butoxycarbonyl)-4-benzyloxy-proline (40.2 mg, 0.000125 M), and benzotriazol-l-yloxytris(dimethylamino)phosphonium-hexafluorophosphate sequalog (55 mg, 0.000125 M) in acetonitrile (3 ml). Diisopropylethylamine (50 ml) wa~ added to make the solution basic. After tlc(silica-methylene chloride(9)-methanol(1)) showed a new spot, the reaction was concentrated to an oil. The oil was dissolved wo 951140~ PCr/US94/13483 ~i ~4~5~

in methylene chloride-ether(l :3) and the solution was washed with water, sodium bicarbonate (sat., aqueous), water, 10% potassium bisulfate, and brin~. After drying with sodium sulfate, the solution was filtered and concentrated to an oil which was purified by "flash" silica 5 chromatography with 40% ethyl acetate in hexane as solvent. Fractions 15-24 contained the product and they were collected and concentrated.
Ether-hexane treatrnent provided a white solid which was the title compound. A 10 ~lg sample was used for characte~ization and testing with the remainder being used in s~lbceq ll~nt steps.

NMR (CDC13) was consistent with structure.
HPLC: 210-99.OQ3% 254-100%
Mass Spectra: Cal'd, 705.965; Found, 706 Analysis calculated for C40HssN306S 0.25 H20 C, 67.62; H, 7.87; N, 5.91 Found: C, 67.56; H, 8.1 1; N, 5.59 ~O2C~
HN N~

Endo-( I S)- I '-(((2-(L-N(tert . -butoxycarbonyl)-4-hydroxy-prolinoylamino)-7,7-dimethylbicyclo(2.2. 1 )-hept- I -yl)methyl-~u If onyl)~piro( I H-indan- I .1 -piperidine) A mix~ure of Endo-(lS)-1'-(((2-(L-N(tert.-butoxy-carbonyl)-4-benzyloxy-prolinoylamino)-7,7-dimethyl)bicyclo(2.2. 1 )-21 7465~

hept- 1 -yl)-methyl)-sulfonyl)spiro( 1 H-indan- 1 ,4-piperidine), ethanol (5 ml), and Pd(OH)2 (60 mg) was hydrogenated on the Parr apparatus at 50 psi for 18 hours. I'he catalyst was removed by filtration and the solvent was removed under reduced pressure to yield the title compound 5 as a white solid.
NMR (CDC13) was cu,lsi~ with structure.
HPLC: 210-93.5% 254-(7.381%
Mass Spectra: ('~lr~ t~-1, 615 840; Found, 616 o Analysis calculated for C33H49N3O6S 1.5 H2O
C, 61.65; H, 8.15; N, 6.53 Found: C, 61.46; H, 7.88; N, 6.90 SO2CH2 H ~>
HN~NH
o Endo-( I S)- I '-(((2-(L-4-hydroxyprolinoylamino)-7,7-dimethyl-bicyclo(2.2. 1 )-hept- I -yl)-methyl)-sulfonyl)spiro( I H-indan- 1,4-piperidine)hydrochloride - A mixture of Endo-(lS)-1'-(((2-(L-N(tert.-butoxy-carbonyl)-~-hydroxyprolinoylamino)-7,7-dimethyl)bicyclo('2.2. 1 )-hept-I-yl)-methyl)-sulfonyl)spiro(lH-indan-1,4-piperidine), was dissolved in ethyl acetate (5 ml) and cooled to -5C. This was placed under a nitrogen atmosphere and hydrogen chloride gas was bubbled in for 10 minutes. The solvent was removed under reduced pressure. Ether was WO 95114025 PC'rlUS94/13483 21 74651~

added and removed under reduced pressure to give 15.48 mg of the title compound as a white solid.
NMR (CDC13) was consistent with structure.
HPLC: 210-93.50% 254-93.812%
Mass Spectra: Calculated, 515.721 (free base); ~ound, 516.4 Analysis calculated for C28H41N34S HCI 0-25 H2O
C, 60.41; H, 7.69; N, 7.54 Found: C, 60.08; H, 7.73; N, 7.35 A~
so2CH2 H
N
H
Endo-( 1 S)- I '-(((2-(3-indolylacetylamino)7 ,7-dirnethyl-bicyclo-(2.2.1)-hept-1-yllmethyl-sulfonyl~spiro(lH-indan-1.4-piperidine) In an oven-dried flask (50 ml) under nitrogen was dissolved endo- I S - I '-(((2-amino-7,7-dimethylbicyclo(2.2. 1 .)-hept - I -yl)-methyl)-sulfonyl)spiro(lH-indan-1,4'-piperidine)(50 mg, 0.000125 M), 3-indolyl acetic acid (21.9 mg, 0.000125 M), and benzotriazol-l-yloxytris(dimethylamino)phosphonium-hexafluorophosphate (Sequalog) (55 mg, 0.0001'25 ~)in acetonitrile (3 ml). Diisopropylethyl~mine (50 ml) was added to mlake the solution basic. After tlc(silica-methylene chloride(9)-methanol(l)) showed a new spot, the reaction was concentrated to an oil. The oil was dissolved in methylene chloride-ether (1:3) and the solution was washed with water, sodium bicarbonate (sat., aqueou.s)~ water, 10% potassium bisulfate~ and brine. After WO 9~/14025 2 1 7 4 ~ ~ ~ PCT/IJS94/13483 drying with sodium sulfate, the solution was filtered and concentrated to an oil which was purified by "flash" silica chromatography with 10%
methanol in methylene chloride as solvent. Fractions 12-19 contained the product and they were collected and concentrated. Ether-hexane 5 treatment provided a white solid which was the title compound.
MR (CDC13) was consistent with structure.
HPLC: 210-99.829% 254-92.699%
Mass Spectra: Calculated, 559.777; Found, 560.3 o Analysis calculated for C33H41N3O3S 0.5 H2O
C, 69.69; H, 7.44; N, 7.51 Found: C, 69.70; H, 7.57; N, 7.20 ~02CH, 7~
O ~ H
Endo-(lS)-I '-(((2-(S-3-(tert.-butoxycarbonyl)amino)-2-oxo-1-pyrro-lidineacetylamino)7 ,7-dimethylbicyclo-(2.2. 1 )hept- I -yl)-methyl-sulfonyl)~piro( I H-indan- I .4-piperidine) In an oven-dried flask (50 ml) under nitrogen was dissolved endo-lS-1'-(((2-amino-7,7-dimethylbicyclo(2.2.1.)-hept-1-yl)-methyl)-sulfonyl)spiro(lH-indan-1,4'-piperidine)(50 mg, 0.000125 M), S-3-((tert.-butoxycarbonyl)amino)-2-oxo-1-pyrrolidine acetic acid (32.2 mg, 0.000125 M), and l,el~uL-iazol-l-yloxytris (dimethylamino)-phosphoniumhexafluoro-phosphate (Sequalog) (55 mg, 0.000125 M) in acetonitrile (3 ml). Diisopropylethylamine (50 ml) wa~ added to make 21 7~0 the solution basic. After tlc(silica-methylene chloride(9)-methanol(l)) showed a new spo~, the reaction was concentrated to an oil. The oil was dissolved in methylene chloride-ether(1:3) and the solution was washed with water, sodium bicarbonate (sat., aqueous), water, 10% potassium 5 bisulfate, and brine. After drying with sodium sulfate, the solu-tion was filtered and ~ C~ lal~d to an oil which was purified by "flash" silica chrornatography with 10% methanol in methylene chloride as solvent. Fractions 15-28 cnnt~inPd the product and they were collected and concentrated. Ether-hexane treatrnent provided a white o solid which was the title compound.
NMR (CDC13) was consistent with structure.
HPLC: 210-94.005% 254-96.33%
Mass Spectra: ('~ tPd m/e, 642.859; Found m/e, 643.4 Analysis c:~ t~dforC34HsoN4O6S"0.1hexane C, 63.79; H, 7.95; N, 8.59 Found: C, 63.~6; H, 8.32; N, 8.29 ~DO2CH~ O

WO 9~/1402~ PCr/US94113483 Endo-(lS)-1'-(((2-(S-3-amino)-2-oxo-1-pyrrolidine-acetylamino)7, 7-dimethylbicyclo(2.2. 1 )-hept- l -yl)-methyl)-sulfonyl)spiro( I H-indan-1 ,4-piperidine)hydrochloride Endo-lS-1 '-(((2-(S-3-(tert.-butoxycarbonyl)amino)-2-oxo-1 -pyrrolidineacetylamino-)-7,7-dimethyl-bicyclo(2.2. 1 .)hept- 1 -yl)-methyl)-sulfonyl)spiro(lH-indan-1,4'-piperidine) was dissolved in ethyl acetate (5 ml) and cooled to -5C. This was placed under a nitrogen atmosphere and hydrogen chloride gas was bubbled in for 10 minutes.
The solvent was removed under reduced pressure. Ether was added and o removed under reduced pressure to give 23.7 mg of the title compound as a white solid.
NMR (CDC13) was ~.",~ "l with structure.
HPLC: 210-96.562% 254-95.493%
Mass Spectra: Calculated for free base m/e, 542; Found m/e, 543.3 Analysis calculated for C2gH42N4O4S " H2O " HCI
C, 58.32; H, 7.59; N, 9.38 Found: C, 58.14; H, 7.97; N, 9.21 so2CH2 HN ~ NH

Step 1: Endo-( I S)- I '-(((2-(L-N-tertbutoxycarbonyl-histidinoyl -amino)-7 ,7-dimethylbicyclo(2.2. 1 )-hept- I -yl)-methyl-~ulfonvl)spiro( I H-indan- I ,4-piperidine) ~ i 14~

In an oven dried flask (50 ml) under nitrogen were dissolved endo-lS-1'-(((2-amino-7,7-dimethylbicyclo(2.2.1.)hept-1-yl)-methyl)-sulfonyl)spiro(lH-indan-1,4'-piperidine) (50 mg, 0.000125 M), L-N-tert, butoxyc~rbonyl histidine (29.9 mg, 0.000125 M), and f benzotriazol-l-yloxytris(dimethylamino)phosphonium-hexafluoro-phosphate (Sequalog) (55 mg, 0.000125 M) in acetonitrile (3 ml).
Diisopropylethylamine (50 ml) was added to make the solution basic.
After tlc(silica-me~ylene chloride(9)-methanol(l)) showed a new spot, the reaction was concentrated to an oil. The oil was dissolved in methylene chloride-ether (1:3) and the solution was washed with water, sodium bicarbonate (sat., aqueous), water, 10% potassium bisulfate, and brine. After drying with sodium sulfate, the solution was filtered and concentrated to an oil, which was purified by "flash" silica chromatography with 10% methanol in methylene chloride as solvent.
Fractions 15-28 contained the product and they were collected and c~ncrntratf~-l Ether-hexane treatment provided a white solid which as the title cornpound.
NMR (CD30D) was consistent with structure.
HPLC: 210-97.822% 254-100%
Mass Spectra: Calculated m/e, 639.859; Found m/e, 640.3 Step 2: Endo-( 1 S)- I '-(((2-(L-N-histidinoylamino)-7,7-dimethyl-bicyclo(2.2. 1 )-hept- I -yl)-methyl-sulfonyl)spiro( I H-indan- I .4-piperidine~
Endo- I S- 1 '-(((2-(L-N-tert.butoxycarbonyl-histidinoyl -amino)-7,7-dimethylbicyclo(2.2.1 .)-hept-l -yl)-methyl)-sulfonyl)-spiro(lH-indan-1,4'-piperidine) (50 mg, 0.000125 M) was dissolved in ethyl acetate (5 ml) and cooled to -5C. This was placed under a 30 nitrogen atmosphe~e and hydrogen chloride gas was bubbled in for 10 minutes. The solvent was removed under reduced pressure. Ether was added and nemoved und~r reduced pressure to give 27.5 mg of the title compound as a white solid.

NMR (CD30D) was consistent with structure.
HPLC. 210-98.652% 254-100%
Mass Spectra: Calculated for free base m/e, 539.7; Found m/e, 540.3 Arlalysis calculated for C29H4lN5o3s 2HC1 2H2 C, 53.69; H, 7.30; N, 10.79 Found: C, 53.58; H, 7.45; N, 11.00 ~ H~O~, o Endo-( l S)- I '-(((2-(D-N-(tert.butoxycarbonyl)-im-benzylhistidino-ylamino)-7,7-dimethylbicyclo(2.2. 1 )-hept- I -yl)-methyl)sulfonyl)-spiro( I H-indan- I .4'-piperidine) In an oven-dried flask (50 ml) under nitrogen were dissolved endo-lS-I '-(((2-amino-7,7-dimethylbicyclo(2.2.1 .)-hept-l -yl)-methyl)-sulfonyl)spiro(lH-indan-1,4'-piperidine) (50 mg, 0.000125 M), D-N-tert. butoxycarbonylim-benzyl-histidine (43 mg, 0.000125 M), and benzotriazol- I -yloxytris(dimethylamino)pho~ i " " ~hf~xafluoro phosphate (Sequalog) (55 mg, 0.000125 M) in acetonitrile (3 ml).
Diisopropyl-ethylamine (50 ml) was added to make the solution basic.
After tlc(silica-methylene chloride(9)-methanol(1)) showed a new spot, the reaction was concentrated to an oil. The oil was dissolved in methylene chloride-ether (1:3) and the solution was washed with water, sodium bicarbonate (sat., aqueous), water, 10% potassium bisulfate, and brine. After drying with sodium sulfate, the solution was filtered and WO 9~;114025 PCINS94/13483 ~7 7~5~

concentrated. Eth~r-hexane provided a white solid which was the title compound.
NMR (CDC13) was consistent with structure.
HPLC: 210-99551%
Mass Spectra: Calculated, m/e 730.Q62; Found, m/e 730.6 Analysis calcul~lted for C41HssNsOsS: 0.65 H20 0.4 hexane C,67.15;H,8.04;N,9.02 Found: C, 67.18; H, 7.73; N, 9.02 Endo-(lS)-I '-(((2-(D-im-benzylhistidinoylamino)-7,7-dimethyl-bicyclo-(2.2. 1 )-hept- I -yl)-methyl)-sulfonyl)spiro( I H-indan- 1,4-25 piperidine)-hydrochloride Endo-~ I S)- I '-(((2-(D-N-(tert.butoxycarbonyl)-im-ben~ylhistidinoylamino)-7,7-dimethylbicyclo(2.2.1 .)-hept- I -yl)-methyl)-sulfonyl)sl~iro(lH-indan-1.4'-piperidine) was dissolved in ethyl acetate (5 ml) and cooled to -5C. This was placed under a nitrogen 30 atmosphere and hydrogen chloride gas was bubbled in for 10 minute.s.
The solvent was removed under reduced pressure. Ether was added ~nd removed under reduced pressure to give the title compound as a white solid.
NMR (CDC13) Wd.S consistent with structure.

-WO95/1402S 21 7 465~ PCr/uss41l34s3 HPLC: 210-100%
Mass Spectra: (~ 1 m/e, 629.879; Found m/e, 630 Analysis calculated for C36H47NsO3S HCI 0.1 ethyl acetate 1.4 H2O
C, 59.33; H, 7.20; N, 9.51 Pound: C, 59.36; H, 7.03; N, 9.50 HN
O N~
O
Endo-(lS)-I '-(((2-(S-3-tert.butoxycarbonylamino-2-oxo-1-ylamino)-7,7-dimethylbicyclo(2.2. 1 )-hept- I -y 1 )-methyl-sulfonyl)~piror I H-indan- I ,4-piperidine) In an oven-dried flask (50 ml) under nitrogen was 2 5 dissolved endo- 1 S- I '-(((2-amino-7,7-dimethylbicyclo(2.2. 1 .)-hept- 1 -yl)-methyl)-sulfonyl)spiro( I H-indan- I ,4'-piperidine)(50 mg, 0.000125 M), S-3-tert.butoxycarbonylamino-2-oxo-1-azepine acetic acid (32.8 mg), and benzotria~ol-l-yloxytris (dimethylamino)phosphonium-hexafluoro-phosphate sequalog (55 mg, 0.000125 M) in acetonitrile (3 ml).
30 Diisopropylethylamine (50 ml) was added to make the solution basic.
After tlc(silica-methylene chloride(9)-methanol(l)) showed a new spot, the reaction was concentrated to an oil. The oil was dissolved in methylene chloride-ether(1.3) and the solution was washed with water, sodium bicarbonate (sat., aqueous), water, 10% potassium bisulfate, and , .. . . . , _ . .

WO 95/14025 PCrlUS94/13483 2 ~ i7 ~

brine. After drying with sodium sulfate, the solution was filtered and concentrated to an oil, which was purified by "flash" silica chromatography with 10% methanol in methy~ene chloride as solvent.
The fractions which contained the product were collected and cnnc~n~ p~ Ether-hexane treatment provided a white solid which was the title compound. A 10 mg sample was saved for characterization and testing.
NMR (CDC13) was Co~ L~ with structure.
o HPLC 210-100%
Mass Spectra calculated m/e 670.918 found m/e 671 Analysis calcul~ted for C36Hs4N4O6S
C, 64.44; H, 8.11; N, 8.35 Found: C, 64.12; H, 8.23; N, 8.03 lS

20 ~ ~
SO2CH2 . H ~NH2 \~ N~j o Endo-(1 S)- I '-(((2-(S-3-amino-2-oxo- 1 -azepine acetyl-amino)-7,7-dimethylbicyclo(2.2. 1 )-hept- I -yl-methyl)-sulfonyl)spiro( 1 H-indan-30 1.4'-piperidine) hvdrochloride Endo( I S)- I '-(((2-(S-3-tert-butoxycarbonyl-amino-2-oxo- 1-azepine acetylamino)-7,7-dimethylbicylo-(2.2. 1 .)-hept- I -yl)-methyl)-sulfonyl)spiro(lH-indan-1,4'-piperidine) was dissolved in ethyl acetate (~ ml) and cooled to 5C. This was placed under a nitro~en atmosphere and hydrogen chloride gas wa~ bubbled in for 10 minutes. The solvent 21 7465û
.

was removed under reduced pressure. Ether was added and removed under reduced pressure to give the title compound as a white solid.
NMR(DCDl3) was Collsi~ t with structure.
HPLC 210-100%
Mass Spectra: Calculated m/e, 570.802; Found m/e, 571 Analysis calculated for C3 lH46N4O4S " HCI 1.10 H2O " 0.40 ethyl acetate C, 59.11; H 7.97, N, 8.46 Found: C, 59.09; H 8.00, N, ~.45 HN ~6~`~
O NH
Endo-(lS)-1'-(((2-(4-imidazoleacetylarnino)-7,7-dirnethylbicyclo-25 (2.2.1)-hept-1-yl)-methyl)-sulfonyl)spiro(lH-indane-1,4'-piperidine)-hydrocloride N~N-bis(2-chloroethyl)-t-butyIcarbamate Di-t-butyldicarbonate (62 g, 0.28 mole, available from 30 Aldrich) and bis(2-chloroethyl)amine hydrochloride (55 g, 0.21 mole ,available from Aldrich) were stirred together in methylene chloride (400 mL). Triethylamine (42 mL, 0.3 mole) was added dropwise but briskly to the stirred suspen.sion. After 10 minutes, additional triethylamine (ca. 5-6 mL) was added to adjust the pH of the mixture (a~s determined by spotting a sarnple on E. Merck pH 5-10 colorpHast sticks, WO 9511402~ PC.rll~S94/13483 2~ ,74~

moistened with water) to 9-9.5. The mixture was stirred for 1 hour at ambient tc~ Jclalu,~7 then filtered. The filtrate was divided iJn half, and each half (ca. 300 mL) was flash chromatographed on a separate 15", 2"diameter silica column packed with 3:2 (v:v) methylene 5 chlori~lf h~Y:ln~ and eluted with 9:1 methylene chloride:hexane. 100 mL fractions were taken, and the product was obtained in fractions ca.
6-12. Assay was b~ TLC on silica gel plates, eluted with methylene chloride and visualized with phosphomolybdic acid stain (Rf ca. 0.75).
Evaporation of the combined product fractions from both columns in vacuo provided the title compound (70 g) as a colorless oil. The oil was twice dissolved in dry THF and evaporated in vaGuo to remove methylene chloride.
I '-(t-Butyloxycarb~nyl)spiro( I H-indene- 1 ,4'-piperidine) To a solution of indene (36.2 g, 310 mmole) in dry tetrahydrofuran (TH~F, 40 mL) cooled in an ice bath and mAinlAin~d under a nitrogen blanket was added (dropped funnel) lithium bis(trimethylsilyl)-amide (620 mL of a 1.0 M solution in THF; 620 mmole, available from Aldrich) over 30 minutes. The mixture was 20 stirred in the cold for 30 minutes, then Llall~rcll~,d by cannula over 20 minutes to a solution of N,N-bis(2-chloroethyl)-t-butylcarbamate (70 g, 290 mmole) in TH[F (40 mL), stirred in an ice bath. The mixture was stirred for 2 hours iln the cold and for 30 minutes at ambient temperature under nitrogen, then evaporated in vacuo to a foam.
25 CH2C12 (400 mL) was added and the resulting mixture divided in half.
Each half was poured onto a silica gel column (15", 2" id) packed and eluted with 1:1 hexane:CH2C12 (2 L) followed by 1:4 hexalne: CH2cl2 The product fractiolls (fractions 3-9 of 200 mL fractions) were evaporated to drynes~ in vacuo to provide l'-(t-butyloxycarbonyl)spiro 30 (indene-1,4'piperidille)~ 90 g total, as a crude yellow solid. The solid was taken up in boiling hexane (400 mL), cooled, and the crystallized solid filtered. The filtrate was repeatedly boiled down to 1/2 its volume, cooled and filtered to obtain successive crops, providing a total 2 ~ 7~6~

of 54 g of pure product. The residue was rechromatographed to provide another 6.7 g (60.7 g total).
Spiro(l H-indene- I ~4'-piperidine) hvdrochloride 1'-(t-Butyloxycarbonyl)spiro(indene-1,4'-piperidine) (60.7 g) in ethyl acetate (700 mL) was stirred in an ice bath and saturated with HCI (g) for 30 minutes, keeping the internal L~ eldlul~ < 12C.
The mixture was stirred in the cold an additional 30 mimutes, then evaporated to dryness. Ethyl acetate was added and removed in vacuo o three times, and the residue was triturated with diethyl ether and filtered to provide spiro(lH-indene-1,4'-piperidine) hydrochloride.
( 1 S)- 1'-(((7 ,7-dimethyl-2-oxobicyclo(2.2. 1 )hept- 1 -)methyl)-sulfonyl)-spiro( I H-indene- 1 ~4'-piperidine) Spiro(lH-indene-1,4'piperidine) hydrochloride (45.4 g, 0.2 mole) and (+)-10-~,a l,~hol~ulfonyl chloride (62.5 g, 0.25 mole, available from Aldrich) were combined in CH2C12 (700 mL) and treated with triethylamine (68.5 mL, 0.5 mole). Additional triethylamine was added as needed to adjust the pH of the mixture to 9-9 5 (moistened E. Merck colorpHast sticks). The mixture was stirred at ambient I~ Jtla~ul~ for 1 hour, then poured onto a silica gel column (10", 2" id) packed with CH2CL2 and eluted with 1:1 Et2o:cH2cl2.
The product fractions were combined and evaporated to dryness in vacuo to provide the title compound as a solid which was recrystallized from petrolium ether and dried 6 hours in vacuo at arnbient lalul~: (m.p. 146-147C).
TLC: Rf=0.44 silica gel (CH2C12).
NMR: Consistent with structure.
HPLC: >99.7% pure.
MS: Molecular ion at m/e - 399 Analysis calculated for C23H2gNO3S
C, 69.14; H, 7.32; N, 3.51 Found: C, 6~.97; H, 7.2; N, 3.38 2~ 7465~ --.

( I S)- 1 '-(((7,7-dime~hyl-2-oximinobicyclo(2.2. 1 )hept- I -yl)-methyl)-sulfonyl)spiro( I H-indene- I .4'-piperidine) (lS)-1'-(((7,7-dimethyl-2-oxobicyclo(2.2.1)hept-1-yl)methyl)sulfonyl)spiro(lH-indene-1,4'piperidine) (30 g, 0.075 mole) 5 in pyridine (500 mL) was heated in an oil bath to 70C (irlternal).
Hydroxylamine hyclrochloride (30 g) was added in three portions over ca. 20 minutes. Af~er 2 hours, an ~l(1itian~l 10 g of hydroxylamine hydrochloride was ~Idded (over 10 minutes). At 30, 40 and 50 minutes ~ltlition~l elapsed time, further 3 g lots of hydroxyl-amine hydrochloride were added. After another 30 minutes, the mixture was poured into water (2 L) and extracted 3X with ethyl acetate (300 mL portions). The organic layers were combined, washed with IN HCI (600 mL tatal), dried over sodium sulfate, filtered, and evaporated to dryness in vacuo. EtOH ~abs; ca. 25Q mL) was added to 15 the resulting thick syrup and the solution allowed to stand at ambient t~ dlUI~ overnight. The mixture was filtered and the filtrate boiled down to ca. 80 mL. After standing, the mixture was again filtered and boiled down to ca. 20 mL. After a third filtration, the filtered solids were combined to give the title compound (28 g).

Endo-(lS)-1'(((2-amino-7,7-dimethylbicyclo(2.2.1)hept-1 -yl)-methyl)sulfonyl)spiro( I H-indane- I .4'-piperidine) Freshly prepared, activated Raney Nickel catalyst (ca. 30 g) in water was allowed to settle and the water decanted. Abs. ethanol 25 (300 mL) was added, and the mixture swirled and again allowed to settle. The solvent ~as decanted. Two more wash-decant cycle~ with 150 mL of ethanol were similarly carried out. (lS)-1'(((7,7-dimethyl-2-oximinobicyclo(2.2. 1 )hept- I -yl)methyl)sulfonyl)-spiro( I H-indene-1,4'-piperidine) (30 g) was ~tirred in a mixture of abs. ethanol (450 30 mL) and 2-methoxyethanol (900 mL), nitrogen was bubbled through the suspension/~olution, and the Raney Nickel catalyst was added. The mixture was hydrogenated under 50 psi overnight- TLC (9:1 CH2cl2:-MeOH. silica gel) showed the reaction to be complete. The catalyst was removed by filtration, and the flltrate evaporated to dryness in vacuo.
.

WO 95/14025 PCrlUS94113483 ~2~ 746~0 The crude solid (27 g) was divided into 7 g batches, and each batch wasdissolved in methylene chloride (ca. 200 mL) and flash chromato-graphed on silica (700 g in a 100 mm column, packed and eluted with

- 8% (v/v) methanol in methylene chloride), taking 200 mL fractions.
5 The exo isomer of the title amine was obtained in fractions ca. 5-7, and the desired endo isomer in fractions ca. 8-16. TLC was on silica, eluted with 8% methanol/methylene chloride, phosphomolybdic acid stain.
The combined product fracbons were evaporated to dryness to provide the title compound (4.5 g from each 7 g lot, ca 18 g total) as a colorless solid.
Endo-(l S)- I '(((2-(4-irnidazoleacetylamino)-7,7-dimethylbicyclo-(2.2.1)-hept-1-yl)methyl)sulfonyl)spiro(lH-indane-1 ,4'-piperidine hydrochloride 4.56 g (11.3 mmols) of Endo-(lS)-1'(((2-amino-7,7-dimethylbicyclo-(2.2.1)hept-1-yl)methyl)sulfonyl)spiro(lH-indane-1 ,4'-piperidine) was dissolved in 50 mL DMF and the solution treated with 2.30 g (14.1 mmols) of 4-imidazole acetic acid, 1.9 g (14.1 mmols) of l-hydroxyl,~l,,ulliazule hydrate (HBT), and 2.7 g (14.1 mmols) of 1-20 ethyl-3-(3-dimethylall-il-op-~yl) carbodiimide HCI (EDC). The pH of the suspension was adjusted to 9.5 with 4.65 mL (33.4 mmols) of triethylamine and the reaction mixture stirred at 25C for 18 hours.
DMF was removed in vacuo and the crude purplish residue treated with water and extracted with EtOAc (3X). The organics were 2s combined, washed with H2O (IX), brine (iX), dried over Na2SO4, filtered and stripped to dryness in vacuo. Flash chromatography of the crude product on silica gel (114/10/1 of CH2cl2lMeoHlconc. NH40H) gave 5.29 g (92%) of desired product as a white foam.
5.17 g (10.1 mmols) of this product was dissolved in 100 30 mL EtOAc. While stirring vigorously, a solution of HCI (g) in EtOAc was added dropwise until precipatation ceased. The slightly gurnmy mixture was stirred 15 minutes at 25C, then evaporated to dryness in vacuo. The residue was restripped 3X from EtOAc, then 3X from WO g511402~ PCr/US94/13483 Et2O. The white solid was scraped from the walls of the flask, t~iturated with Et2O and 5.3 g of hydrochloride salt collected.
M.P.: 93-167C (slow fo~m) 5 HPLC: 99.4%
PMR: (~nn~i~t~nt with structure, plus 0.30 ethyl acetate, 0.05 ether and H2O.
M.S.: M+H + 511 ~FAB) Analysis calculated for C2gH3gN4O3S HCI 0.30 C4Hgo2 0.05 C4H1oO " 0.4 H2O. (M.W. 584.48) C, 60 41; H, 7.36; N, g.59 Found: C, 60.38; H, 7.46; N, 9.33 ~XAMPLE 23 SO2CH2~_ Br~(~
\/
I -[[6-bromo- 1 ,7-dimethyl -2-oxobicyclo[2.2. 1 ]hept-7-yl)methyl] -sulfonyll-spirol IH-indene-l .4'-piperidinel Dissol~ed 100 mg of (+)-3-Bromocamphor-sulfonic acid 30 ~mmnnillm salt (.304 mM) in 15 mL of DLM. Added 5 eq of SOC12 (FW=I 18.97; d=1.631; 1.52 mM; 120 mL). The mixture was allowed to react overnight, l~hen concentrated to obtain 92.7 mg of the crude sulfonyl chloride. The 92.7 mg of sulfonyl chloride was dissolved in 25 mL of dry THF. T~ this was added 1.1 eq of indene salt (68.59 mg) WO 95/14025 PCI`/US94/13483 2l~74650 and 2 eq of TFA, which was allowed to react at room temperature for - three hours. The product was concentrated and washed with HCI, H2O
and brine. Flask chromatography in 20% EtOAc/petroleum ether.
HPLC 97.7% at 13.52 min.

2~' 20 (IR-syn)-1'-[[(1,7-dimethyl-2-oxobicyclo[2.2.1]hept-7-yl)methyl)-sulfonyll -spiror I H-indene- I .4'-piperidinel To a solution of the product of Example 23 (107 mg, 0.300 mmol) in glacial acetic acid (10 mL) was added zinc dust (24 mg, 0.367 mmol). The temperature was then increased to reflux. After 30 25 mmutes, the mixture was cooled to room temperature, filtered, then concentrated under reduced pressure. Purification by flash chromatography (35% ethyl acetate in petroleum ether as eluent) afforded P~0 mg of the product as a white amorphous foam.
NMR (300 MHz, CDC13): Consistent with structure.
HPLC (Vydac C18 column, gradient from 95/5 to 0/100 H20/CH3CN
with 0.1% TFA, 15 min. gradient, flow rate = 1.5 mL/min): purity =
98%, r.t. = 12.67 min.
FABMS: [M+l]+] at400.91 Analysis calculated for C23H2gNO3S
i wo ssrl402s PcrNss4ll3483 .
2~ i 4~5 C, 69.13; H, 7.32; N, 3.51 Found: C, 69.25; H, 7.30; N, 3.48 E~XAMPLE 25 `so2c~-~\D//o o,3~-CF3 o (IS)-1'(((2-(trifluoromethyl)sulfonyl)oxy)2,3-ene-7,7-dimethyl-bicyclo(2.2. 1 )hept- I -yl)methyl)sulfonyl)-spiro(l H-indene- 1,4'-piperidine) To a solution of (IS)-1(((7,7-dimethyl-2-bicyclo(2.2.1)-hept-l-yl)methyl)sulfonyl)spiro(lH-indene-1,4'-piperidine) (2g, 5.0 20 mmol) and 2,6-di-tert-butyl-4-methylpyridine (1.5 g, 7.5 mmol) in CH2C12 (25 ml) was added triflic anhydride (1.3 ml, 7.5 mmol) and the mixture was stirrea at room L~ e-a~u-~ for 30 minutes. The mixture was then diluted with CH2C12 (30 ml) and filtered. The filtrate was then washed with 5% HCI (2x50 ml), saturated NaHCO3 (2x50 ml), and 25 brine, dried over Na2SO4, and evaporated.
The crude triflate was purified by flash chromatography eluting with 20% et~lylacetate in hexanes to yield 1.7 g of the title product as a white foam (64%).
3 0 HPLC RT = 12.75 min.
NMR (CDC13) in agreement with title compound.
FAB MS: 532 (M+l) Analysis calculat~d for C42H2gNS2OsF3 N, 2.63. C, 54.22, H, 2.63 Found: N, 2.37: C, 54.~2; H, 5.34 2 1 7 ~ 6 ~ 0 5 1~
N\ /~1 ~0 H3CO \

( 1 S)- I '-(((2-(dimethylphosphonyl)oxy)2,3-ene-7 ,7-dimethyl-yc1O(2.2.1)hept-1-yl)sulfonyl)spiro(lH-indene-1.4'-pipendine) A mixture of the triflate product of Example 25 (50 mg, 0.097 mrnol), dimethyl phosphite (13 ml, 0.14 mmol), triethylamine (61 ml, 0.44 mmol), and tetrakis(triphenylphosphine)r~ lm (5 mg, 0.005 mmol) in DMF (3 ml) was stirred under an atmosphere of argon 20 for I hour. The reaction was then diluted with CHC13 (25 ml). The chloroform was washed with 5% HCI (2x25 ml) and brine, dried over Na2SO4 and evaporated to dryness. Purification via flash chroma-tography (25% ethylacetate in hexanes) afforded 40 mg of the title compound as a white solid (~6%).
HPLC Rl` = 10.31 min.
NMR(CDC13) in agreement with title compound FAB MS: 492 (M+l) Analysis calculated for C2sH34NSOsP 0.5 H2O
~o N, 2.80; C, 59.99; H, 7.05 Found: N, 2.71; C, 59.90; H, 7.49 wo 95/14025 PCr/US94113483 21 ,~G~i~

so2cH2~?
N ~ O/~
S~
[l-[[[[exo-2-hydroxy-777-dimethyl-l-[spiro[lH-indene-l74-piperidin]-I -yl-sulphonyl)met 1yl]bicyclo]2.2. 1 ]hept-2-yl]methyl]amino]carbonyl]-3-(methvlthio)propyll-carbamic acid-l.l-dimethylethyl ester To a stirred solution of (IS)-I'-(((exo-2-hydroxy-endo-2-:~min~m!~thyl-7,7-dimethylbicyclo(2.2.1)hept-1-yl)methyl)sulfonyl)-spiro(lH-indene-1,4'-piperidine) (600 mg, 1.39 mmole) in 5 ml of dry, degassed N,N-dimethylformamide was added Boc-L-methionine (381 mg, 1.53 mrnole) and 743 mg (1.68 mmole) of benzotriazol-l-yloxy tris(dimethylamino)phosphonium hexafluorophosphate (BOP) at room temperature. The resulting reaction mixture was protected from moisture and the p~ was adjusted to 8-9 with diusopropylethylamine.
After one hour, all volatile components were removed under reduced pressure and the residue was dissolved in 250 ml of ethyl acetate. This 601ution was washed in succession with 10% aqueous citric acid, 50%
sodium bicarbonate solution, and brine. The organic phase was dried (sodium sulfate) ani concentrated. Chromatography of the crude reaction product on silica gel (1:1 ethyl acetate-hexane elution) afforded the title compound as an amorphous solid: m.p. 82-92C.
HPLC: >92% pure at 214 nM
NMR: Consistent ~vith structure and verified presence of solvent WO 95/14025 PCr/lJS94/13483 21 74~

FAB MS: 670 (M+ + thiogIycerol) Analysis calculated for C34Hs1N3O6S2 O.5CHC13 C, 57.42; H, 7.19; N, 5.82 Found: C, 57.58; H, 7.50; N, 5.83 ~r~
- OH o H~S~
' 2-amino-N-[[exo-2-hydroxy-7,7-dimethyl- 1 -[(spiro[lH-indene-l ,4'-piperidin] I 'ylsulfonyl)methyl]bicyclo]2.2. 1 ]hept-2-yl]methyl] -4-methylthio)-butanamide A c-~ntinl~o~l~ stream of dry HCI gas was passed for S
minutes into an ice cold solution of ethyl acetate (2 ml) containing 20 mg of the product of Example 27. After 30 minutes at 0C, the reaction mixture was cu~ ai~d and the residue was chromatographed on two 0.25 mm precoated silica gel plates (chloroform-methanol-ammonium hydroxide, 93:7:0.7 v/v elution). The title compound was obtained as a solid (10.7 mg): m.p. 78-80"C.
TLC: Rf=0.28 (CHC13-CH30H-NH40H, 95:5:0.5) NMR: Consistent with structure and verifies presence of solve FABMS: 562(M++H) Analysis calculated for C2gH43N3O4S2âO.75CHCI-~
C, 54.86; H, 6.77; N, ~.45 Found: C, 54.52; H, 6.90, N, 6.3Z

~ 7 ~

s 0 ~OH~ ~
5-(1,3-dihydro-1,3-dioxo-2H-isoindol-2-yl)-N-[~2-hydroxy-7,7-dimethy- I -[(spiro[ I H-indene- I ,4'-piperidin] - I '-ylsulfonyl)methyl] -bicyclo[2.2.1]hept-2-yl]-methyl]dihydro-4-oxo-2H-1,3-Thiazine-3(4H)-~et~m j~l~
To a slirred solution of (lS)-I'-(((exo-2-hydroxy-2-20 aminomethyl-7,7-di~ llyl~icyclo(2.2.1)hept-1-yl)-sulfonyl)spiro(lH-indene-1,4'-piperidine) (67 mg, 0.16 mmole) in 1.5 ml of dry, degassed N~N-dilll~lllylru~ dlllide was added 4-oxo-~-phthalyl-1,3-thiazine-acetic acid (62 mg, 0.192 mmole) and 88 mg (0.20 mmole) of l~ell~ullia~
yloxytris(dimethylamino)-phosphonium hexafluorophosphate (BOP) at 2s room l~ ,eldlul~;. The resulting reaction mixture was protected from moisture and the pH was adjusted to 8-9 with diisopropylethylamine.
After 24 hours all volatile components were removed under reduced pressure and the residue was suspended in 5 ml of toluene.
Cnn~ntr~tion of this suspension afforded the crude product which wa.
30 chromatographed on silica gel (96:4:0.4 CHC13-CH30H-NH40H) to give the title compcund (109 mg): m.p. 134-137 C.
NMR: Consistent vvith structure and verifies presence of solvent;
HPLC: >94% pure at 214 nM;
FAB MS: 733 (M+~H) 21 7465~

Analysis calculated for C3gH44N4O7S21.1CHC13 C, 54.34; H, 5.26; N, 6.48 Found: C, 54.12; H, 5.22; N, 6.42 H ~H2 5-arnino-N-[[1 -[[spiro[lH-indene-l ,4'-piperidin]-1-yl)-sulfonyl]-methyl]-exo-2-hydroxy-7,7-dimethylbicyclo-[ 1.1. I ]hept-2-yl]-methylldihvdro-4-oxo-2H- I .3-Thiazine-3(4H)-acetamide The product of Example 29 (60 mg) was dissolved in 3 ml of methanol and treated with 50 ml (1.59 mmole) of 95% hydrazine at 23C. After 24 hours the solvent was removed under vacuum and the residue was suspended in toluene. Rotoevaporation of this suspension gave the crude product, which was chromatographed on four 0.5 mm procoated silica gel plates (chloroform-methanol-ammonium hydroxide, 92:8:0.~ v/v elution). The title compound was obtained as an amorphous solid (27 mg): m.p. 103-106C.
NMR: Consistent with structure and verifies presence of solvent HPLC: >97% pure at 214 nM;
FAB MS: 605 (M++H) Analysis calculated for C30H44N4OsS2 0.6CHC13 C, 54.33; H, 6.64; N, 8.28 Found: C, 54.12; H, 6.60; N, 8.20 WO 9!i11402~ PCT/US94/13483 21,7~

E~XAMPLE 31 5 g3 N ~/--\SO2 ~

1 '-[(5-oxospiro[bicyclo[2.2.1]heptane-7,1 '-cyclopro-pan]-2-yl)-sulfonyll-spirorlH[-indane-l ~4'-piperidinel Spiro( I H-indane- I ,4'-piperidine)hydrochloride(3 g, 13 .S
mmole) was dissolved in 30 ml of methylene chloride. The resulting solution was cooled to 0C and treated with 3.76 ml of triethylamine.
This was followed by the dropwise addition of chloroethyl-sulfonylchloride (1.57 ml, 15 mmole). The reaction mixture was allowed to warm to room temperature over one hour and was filtered.
The filtrate wa~ washed with water~ sodium bicarbonate solution, and brine. The dried organic washings were ~ullct;llllal~d and the residue was column chromatographed on silica gel (98:2 methylene chloride-ether elution) to give 1.88 g of l'-(vinylsulfonyl)spiro(lH-indane-1,4'piperidine)(m.p. 114-115C).
I '-(Vinylsulfonyl)spiro( I H-indane- 1 74'piperidine) ( 185 mg, 0.67 mmole) was combined with 5.1 g (55 mmole) of ~ ulle~diene in 4 ml of benzene arld heâted to reflux for 48 hours. The solvent and excess reagent were removed under reduced pressure and the residual ûil was chrûmatûglâphed on ~ilica gel (4:1 hexâne-ethyl âcetate elution) to give 216 mg of a solid which was recrystallized from methanol to give white needles (m.p. 176-177C). Of this material, 110.7 mg was dissolved in 5 ml of dry THF under nitrogen. This solution was cooled to 0C and treated with 0.1 ml of borane-THF complex. After stirring for one hour, an additional 0.1 ml of Borane-THF complex was added to the reaction mi~;ture and stirring was continued at room temperature for I hour more. The reaction mixture was recooled to 0C and WO 95114025 2 ~ 7 ~ 6 5 0 PCIIUS94/13483 quenched with the dropwise addition of water. After 10 minutes, 0.3 ml of 3N sodium hydroxide solution and 0.3 ml of 30% hydrogen peroxide solution was added. The resulting mixture was heated to ~0C
for 1 hour and allowed to stand at 23C overnight. The reaction 5 mixture was partitioned between ethyl acetate and brine. The aqueous phase was extracted with ethyl acetate and the combined organic extracts were dried (MgSO4) and concentrated. Purification of the crude reaction product by preparative TLC on silica gel (2:1 hexane-ethyl acetate) afforded two coll,po,l~llL~. The more polar product (m.p. 232-o 233C, from methanol) (0.15 mmole) was dissolved in 5 ml ofmethylene chloride and oxidized with pyridinium chlorochromate (2 mmole) at 23C. The reaction mixture was filtered through celite and the filtrate was applied to 0.25 mm precoated silica gel plates. Elution with 2:1 hexane-ethyl acetate and recrystallization from methanol 5 afforded the title compound as a white solid: m.p. 210-212C.
NMR: Consistent with structure and verifies presence of solvent HPLC: >96% pure at 214 nM;
FAB MS: 386 (M++H) Analysis calculated for C22H27NO3so 3cH2o C, 67.59; H, 7.12; N, 3.58 Found: C, 67.66; H, 7.22; N, 3.53 WO 95/14025 PCI'NS94/13483 ~ l 4f~5f~

I '-[(g, I 0-dihydro-~, 1 0-ethanoanthracen- 1 ] -yl)sulfonyl] -spiro[ I H-indane- I .4'-pipericlinel I'-(Vinylsulfonyl)spiro(lH-indane-1,4'piperidine) (152 mg) was combined with 350 of anthracene m 15 ml of toluene and 5 heated to reflux for 8 days. The solvent and excess reagent was removed under reduced pressure and the residual oil was chromatographed on silica gel (4:1 hexane-ethyl acetate elution) to give the title compound as an amorphous solid.
o NMR: Consistent ~vith structure and verifies presence of solvent HPLC: >99% pure at 214 nM;
FAB MS: 456 (M~+H) Analysis calcul~ted for C2gH2gNO2S0.05C H2O 0.25 HC13 C, 72~3; H, 6.08; N, 2.88 Found: C, 72.23; H, 5.92; N, 2.67 20 l~ ~
SO2~
I '-(spiro[bicyclo[2.2. 1 ]hept-5-ene-7,1 '-cyclopentan]-2-ylsulfonyl)-spiro~ 11 I-indane- I l ~'-piperidinel l'-(Vinylsulfonyl)spiro(lH-indane-1,4'piperidine) (100 mg) was combined with 500 mg of spirononadiene in 3 ml of toluene 30 and heated to reflux for 17 hours. The solvent and excess reagent were removed under reduced pressure and the residual oil was chromato-graphed on silica g~l (4:1 hexane-ethyl acetate elution) to give the title compound a~ an amorphous solid: m.p. 115-119C.
NMR: Consistent with structure and verifies presence of ~olvent WO 95/14025 2 1 7 4 ~ 5 0 PCr/US94/13483 HPLC: >99% pure at 214 nM;
FAB MS: 398 (M++H) Analysis ~lC~ d for C24H31N02SO.05CHC13 C, 71.57; H, 7.76; N, 3.47 Found: C, 71.94; H, 7.81; N, 3.42 ~
52/~
I '-(bicyclo[2.2. 1 ]hept-5-en-2-ylsulfonyl)spiro[ 1 H-indane- 1,4'-piperidinel I '-(Vinylsulfonyl)spiro( I H-indane- 1 ,4'piperidine) ( l O0 mg) was combined with ten equivalents of cyclohPY~ n-~ in 10 ml of toluene and heated to reflux for 30 hours. The solvent and excess reagent were removed under reduced pressure arld the residual oil was chromagraphed on silica gel (4:1 hexane-ethyl acetate elution) to give the title compound as an amorphous solid: m.p. 162-166C.
NMR: Consistent with structure and verifies presence of solvent HPLC: >99% pure at 214 nM;
FAB MS: 358 (M++H) Analysis calculated for C21H27N02SO.lH20 C, 70.19; H, 7.63; N, 3.90 Found: C, 70.30; H, ~.03; N, 3.61 WO 95114025 PCrlUS94113483 .
2~ 7~55 E~AMPLE 35 `o~O~OCH3 1'-[[2'-[[(2-methoxyethoxy)methoxy]methyl]spiro]bicyclo[2.2.1]hept-5-ene-7.1'-cyclopropanl-2-yllsulfonyll-spirorlH-indane-1 .4'-piperidinel 1 '-(Vinylsulfonyl)spiro( 1 H-indane- 1 ,4'piperidine) (2.45 g) was combined with 2.2 g of hydroxymethylspirocycloheptadiene and 3 mg of hydroquinone in 8 ml of toluene and heated to reflux for 17 5 hours. The solven~ and excess reagent were removed under reduced pressure and the residual oil was chromatographed on silica gel (1:2 hexane-ethyl aceta~e elution) to give three components. The least polar of these products (80 mg) was dissolved in methylene chloride containing 78 mg of diisopropylethylamine and treated with 49 mg of 2-20 methoxyethoxy-m~.thyl chloride. The reaction mixture was protected from moisture and stirred at 23C overnight. The reaction mixture wa~
conl ~ntr~t~d and tile residual material was applied to 0.5 mm precoated silica gel plates. Elution with 1:1 hexane-ethyl acetate afforded the title compound.

NMR: Consisten~ with structure and verifies presence of solvent;
PLC: >90% pure at 214 nM;
FAB MS: 4RR (M~ +H) wo 95/14025 PCr/US94113483 21 7465~

E~XAMPLE 36 ~ /
\NJ~N~co2cH3 (lS-)-2-[[[exo-2-hydroxy-7,7-dimethyl-1-[(spiro[lH-indene-1 ,4'-piperidin]-l '-ylsulfonyl)methyl]bicyclo[2.2.1]hept-2-yl]methyl]amino]-N-(2-methoxy-2-oxoethyl)-N,N-dimethyl-2-oxo-~1l"."~."i"iu"l salt with trifluoroacetic acid (1:1) To a stirred solution of (lS)-I'-(((exo-2-hydroxy-endo-2-aminomethyl-7,7-dimethylbicyclo(2.2. 1 )hept- I -yl)methyl)sulfonyl)-spiro(lH-indene-1,4'-piperidine) (108 mg, 0.25 mmole) in 5 ml of 20 toluene was added I ml of toluene containing 0.5 mmole of methyliminodiacetic acid anhydride. Tetrahydrofuran (2 ml) was added and the white suspension was stirred at 23C overnight. The reaction mixture was filtered and concentrated. The residue was dissolved in 5 ml of N,N-dimethylformamide and treated in succession ~vith methyl 2s iodide (0.5 ml) and (liisoplu~ylethylamine (0.2 ml). The resulting solution was protected from moisture and heated for 5 hours at 50C.
An additional 0.5 ml of methyl iodide was added and heating was continued for 5 hours more. The reaction mixture was concentrated and the residue was purified via reverse phase preparative HPLC
30 (Vydac Protein & Peptide C-18 column, mobile phase = 0.1% trifluoro-acetic acid (TFA) in water-acetonitrile). The fractions containing the title compound were pooled and c~ elltldl~d. The residue was dissolved in dioxane and this solution was freeze-dried to yield the TFA
salt of the title compound as a white powder:

~Vo 9~114025 PCr/US94113483 .
2~ 74~53 NMR: (~nnsi~t~nt with structure and verifies presence of solvent HPLC: >99% pure at 214 nM;
FAB MS: 588 (M~-+H) Analysis calcul~lted for C31H46N3O6S 1.8CF3CO2H 1.0 dioxane C,52~1;H,6.27;N,4.77 Found: C, 52.~8; H, 6.40; N, 4.76 [~
S2~
Br>~/ l\
0~

(lR-syn)-1'-[[(1,7-dimethyl-2-oxobicyclo[2.2.1]hept-7-yl)methyl]-sulfonvll-spiro~ lH-indene-l ~4'-piperidinel Disso~ved 100 mg (.304 mM; FW=328.23) of (-)-3-brom~- ~mrhosulfonic acid ~mm~nillm salt in 15 mL of DCM. Added S
25 eq of tllionylchloride (FW=I 18.97; d=1.631; 1.52 mM; 120 mL) and allowed mixture to react at 0C under a N2 balloon for I hour. After I
hour the mixture ~as concentrated to obtain 63.12 mg of crude acid chloride (.191 mM).
Dissolved 47.13 mg of indene HCI salt in 20 mL of THF.
30 The acid chloride ~vas added along with 2 eq of DEA, and allowed to react at room temperature for I hour, then concentrated and washed with~x~00mLofI NNCI,2x200mLH2Oand2x200mLbrine, then concentrated and dried over sodium sulfate. Flash chromatography in 25% EtOAc/petroleum ether.

wo 95114025 PCrlUS94/13483 2~ 74~50 N
O î~
~/~
To a solution of the product of Example 37 (107 mg, 0.300 5 mmol) in glacial acetic acid (10 mL) was added zinc dust (24 mg, 0.367 mmol). The temperature was then increased to reflux. After 30 minutes the mixture was cooled to room ~ yeldtu-~, filtered, then ~u-~ d under reduced pressure. Purification ~y flash chromatography (35% ethyl acetate in petroleum ether as eluent) 20 afforded 80 mg of the product as a white amorphous foam.
NMR (300 MHz, CDC13): Consistent with structure;
HPLC: (Vydac C18 column, gradient from 95/S to 0/100 H20/CH3CN
with 0.1% TFA, 15 min. gradient, flow rate = 1.5 mL/min.):
25 Purity: 98%, r.t. = 12.67 min.
FAB MS: [M + l]+ at 400.91 Analysis calculated for C23H2gNO3S
C, 69.13; H, 7.32; N, 3.51 Found: C, 69.25; H, 7.30; N, 3.48 ~o WO 95/14025 PC~r/US94113483 21 7~650 N

I '-[[(7,7-dimethyi-.~-oxobicyclo[2.2. 1 ]hept-l -yl)-methyl]sulfonyl]-3'-phenvl-spiror I H-indene- I .4'-piperidinel Ethanolamine (6.66 mL, 0.1103 mmol) was placed in a 3-neck round bottom flask and heated to reflux. Once at relux, styrene oxide (6.28 mL, 0.0557 mmol) was added dropwise to the reaction mixture over 15 m~nutes. The reaction mixture was refluxed an additional 2 hours and then allowed to cool. The product, N-(2-hydroxyethyl)-N-(2-hydroxy-2-phenylethyl)amine, was obtained as a clear oil by vacuunl distillation.
N-(2-hydroxyethyl)-N-(2-hydroxy-2-phenylethyl)amine (1.30 g, 7.16 mmol) was dissolved in chloroform and then while under a nitrogen blanket, thionyl chloride (1.05 mL, 14.32 mrnol) was added dropwise over 10 minutes. The reaction mixture was then refluxed for I hour and allowed to cool to room temperature. Water was then added 25 to the mixture and allowed to stir I hour. The reaction mixture was separated and the organic layer was washed three times with 3N HCI.
The aqueous layers were combined and then basified with 40% sodium hydroxide. The aqueous layer was extracted 3 times with ether, the organics were combined, and dried over sodium sulfate. The organic 30 layer was filtered and the filtrate concentrated to a yellow oil.
The yellow oil was dissolved in ether and then di-t-butyl dicarbonate (1.50 g, 6.87 mmol) was added along with triethylamine (500 ml, 3.59 mmol).The mixture was allowed to stir overnight and then it was washed with H2O (2x), 0.1 N HCI (2x) and sodium bicarbonate (2x). The organic layer was dried over sodium sulf~te, 21 7455i3 filtered, and the filtrate concentrated to a yellow oil. The yellow oil was dissolved in CH2C12 and poured onto a silica gel column. The column was eluted with 1:1 CH2C12/hexane. The product fractions - were combined and concentrated to dryness to yield the product, N-(2-5 hydroxyethyl)-N-(2-hydroxy-2-phenylethyl)-t-butyl-carbamate.
To a solution of indene (268 ml, 2.30 mmol) in dry THF (2 mL) cooled in an ice bath and m~int~in~d under a nitrogen blanket was added lithium bis(trimethylsilyl)amide (IM solution in THF, 4.6 mL, 4.60 mmol) over 10 minutes. The mixture was stirred in the cold bath o for 30 minutes, then added over 10 minutes to a solution of N-(2-hydroxyethyl)-N-(2-hydroxy-2-phenylethyl)-t-butylcarbamate (694 mg, 2.30 mmol) stirred in an ice bath. The mixture was stirred for 2 hours in the cold and for 30 minutes at 25C under nitrogen, then concentrated to an orange oil. 10% ethyl acetate in hexane was added 15 and the resulting mixture poured onto a silica gel column packed with 10% ethyl acetate in hexane. Elution was with the sarne solvent and the product fraction were cnncçntr~t~d to provide l'-(t-butyloxycarbonyl)-spiro-(indene-3 '-phenyl - I ,4'-piperidine .
1 '-(t-Butyloxycarbonyl)spiro(indene-3-'phenyl- 1,4'-20 piperidine) (290.9 mg, 0.842 mrnol) in ethyl acetate was stirred in anice bath and saturated with HCI (g) for 30 minutes. The mixture was concentrated to dryness and reconcentrated from ether three times to yield spiro ( I H-indene-3 -phenyl- I ,4'-piperidine)-hydrochloride.
Spiro( I H-indene-3'-phenyl- 1 ,4'-piperidine)hydrochloride 25 (205.2 mg, 0.690 mmol) and (+)-10-camphorsulfonyl chloride (196 mg, 0.782 mmol) were combined in THF and the pH was adjusted to 9 with triethylarnine (196 mL, 1.41 mmol). The reaction mixture was stirred ovemight at 25C, then concentrated to dryness, and redissolved in CH2C12, then poured onto a silica gel column and eluted with CH2C12 30 The product fractions were combined and evaporated to dryness to provide the title compound which was crystallized from ether and dried in vacuo overnight.
m.p.: 1~3 - 215C

WO 95~14025 PC~r/US94/13483 2~ i74~

NMR: Consistent with structure HPLC: >95% pure MS: M + H 476.3 (FAB) Analysis calcul~lted for C29H33N03S SOH20 C, 71.~6; H, 7.07; N, 2.89 Found: C, 71.~R; H, 7.03; N, 2.87 ~
SO2 y N,LO
H

(I S(la,2a,4a))-2-h~droxy-7,7-dimethyl-1 -((spiro-(lH-indene,l ,4'-yiperidin)- I '-yl-sull-onvl)methyl)-bicyclo-(2.2. 1 )heptane-2-acetic acid (100 mg, 0.217 mmol) diphenylphosphoryl azide (51.~ mL.
0.239 mmol), and triethylamine (66.2 mL, 0.471 mmol) were combined in DMF. The mixtllre was allowed to stir overnight and then it was concentrated to dryness. The resulting residue was dissolved in CH2C12, poured onto a silica gel column, and eluted with 5% methanol in CH2C12. The product fractions were combined and concentrated to dryness. The title compound was obtained as a white solid from ether and dried in vacuo overnight.
NMR: Consistent with structure HPLC: >93% pure MS: M + H 4~7.3 (FAB) Analysis caiculated for C2sH32N2O4S
C, 6~.76; H, 7.06; N, 6.14 Found: C, 65.76; H, 7.42; N, 5.80 .

wo g5,l4025 2 1 7 4 6 5 0 PCT/IJS94/13483 ~2 - ~ /
H ~
(lS-exo)-[[2--hydroxy-7,7-dimethyl-1 -[(spiro[lH-indene-1,2'-piperidin)-1'-ylsulfonyl)methyl]bicyclo2.2.1]hept-2-yl]carbarnic acid l.l-dimethylethyl ester ( 1 S)- I '-(((7,7-dimethyl-2-oxobicyclo-(2.2. 1 )-hept- I -yl)-methyl)sulfonyl)spiro(lH-indene-1,4'-piperidine) (1.92 g, 4.81 mmol) was combined with zinc iodide (47 mg, 0.15 mmol) in 2 mL of toluene.
While under a nitrogen atmosphere, the mixture was treated with trimethylsilyl cyanide (960 mL, 7.21 mmol) dropwise. The reaction mixture was then heated to 100C for 3.5 hours. The mixture was allowed to cool and diluted with 10 mL of dry THF. Then lithium ~ min~lm hydride (IM in THF; 8.6 mL, 8.6 mmol) was added dropwise over 5 minutes and the mixture was allowed to stir at 25C for 2 hours.
The reaction mixture was then diluted with ether (N50 mL) and 10%
sodium hydroxide solution was added dropwise until a gray precipitate stopped forming. The mixture was now filtered, the filtrate was washed with sodium bicarbonate and brine, the organic layer was separated and dried over sodium sulfate. The or~anic layer was filtered, the filtrate concentrated and the residue dissolved in CH2cl2. This was poured onto a silica gel column, eluted with 97/3/0.3 of CH2C12/methanol/
ammonium hydroxide and product fractions collected. The product fractions were combined and concentrated to dryness. The desired product, ( I S~- I '-(((~,7-dimethyl-(2-endo-aminomethyl-2-exo-hydroxy)-WO gS~14025 PCrlUS94113483 .
21 ~465G

bicyclo-(2.2.1)-hept-1-yl)-methyl)-sulfonyl)spiro(lH-indene-1 ,4'-piperidine), was obtained as a white foam from ether.
(I S)- I '-(((7,7-dimethyl-(2-endo-aminomethyl-2-exo-hydroxy)-bicycylo (2.2.1)-hept-1-yl)-methyl)-sulfonyl)spiro(lH-indene-1,4'-piperidine) (44.9 mg, 0.104 mmol) and di-t-butyl dicarbonate (25 mg, 0.115 mmo~) were combined in S mL of CH2cl2. The reaction mixture was treate~3 with triethylarnine (18.1 mL, 0.13 mmol) and then stirred for 30 minutes at 25C. The reaction mixture was poured onto a silica gel column and eluted with 15% ethyl acetate in hexane. The 0 product fractions were combined and evaporated to dryness. The title compound was obtained as a white solid from CH2C12/hexane and was dried in ~asl overnight.
m.p.: 71C-1 15C
NMR: Consistent with structure HPLC: >95% pure Analysis calculated for C2gH42N2OsS 0.45 C, 6~I4; H, 8.73; N, 5.03 Found: C, 66.r'1; H, 8.52; N, 4.75 N r \SO2 H o OH
N~
H /
~`OH

WO 9~/14025 PCr/Usg4113483 21 74~

(lS-exo)-3-hydroxy-N-[[2-hydroxy-7,7-dimethyl-1-[(spiro[lH-indene-1 ,4'-piperidin]- 1 '-ylsulfonyl)methyl]bicyclo[2.2. 1 ]hept-2-yl]methyl] -2-(hvdroxymethyl)-2-methyl-propanamide (lS)-1 '-(((7,7-dimethyl-(2-endo-aminomethyl-2-exo-5 hydroxy)-bicicylo-(2.2. 1 )-hept- I -yl)-methyl)-sulfonyl)spiro( 1 H-indene-1,4'-piperdine) (51 mg, 0.118 mmol), 2,2-bis(hydroxymethyl)-propionic acid (19 mg, 0.142 mmol), 1-hydroxybenzotriazole hydrate (HBT) (19 mg, 0.142 mmol), and 1-ethyl-3-(dimethylaminopropyl) carbodiimideâHCl(EDC) (27 mg, 0.142 mmol) were all combined in 5 o mL of DMF. Then, triethylamine (46 mL, 0.33 mmol) was added to adjust the pH to 9, and the mixture was allowed to stir overnight at 25C. The reaction mixture was concentrated to dryness and treated with 5% citric acid solution. The water layer was then basified with saturated sodium bicarbonate and extracted 3 times with ethyl acetate.
5 The organics were combined and dried over sodium sulfate. The organic layer was filtered, the filtrate concentrated, and the resulting residue dissolved in CH2C12. This was then poured onto a silica gel column and eluted with 3% methanol in CH2cl2. The product fractions were combined and evaporated to dryness. The title compound was 20 obtained as a white solid from CH2cl2thexane and was dried in vacuo overnight.
m.p.: 167-170C
NMR: Consistent with structure Z5 HPLC: >99% pure MS: M + H 547.3 (FAB) Analysis calculated for C2gH42N2O6S
C, 63.71; H, 7.74; N, 5.12 Found: C, 63.65; H, 7.59; N, 4.88 wo 95/14025 PCr/usg4ll3483 2 1 7~50 - OH
,~
(lS-exo)-4-benzoyl-N-[[2-hydroxy-7,7-dimethyl-1-[(spiro[lH-indene-1,4'-piperidin]-1'-ylsulfonyl)methyl]bicyclo[2.2.1]hept-2-yl]methyl]-benzamide The procedure of Example 42 was carried out using 49.6 mg, 0.115 mmol of (lS)-(((7,7-dimethyl-(2-endo-aTninomethyl-2-exo-hydroxy)-bicicylo-(2.2.1)-hept-1-yl)-methyl)sulfonyl)spiro(lH-indene-1,4'-piperidine),26.~ mg, 0.138 mmol of EDC, 18.7 mg, 0.138 mmol of HBT, 48 mL, 0.345 rnmol of triethylamine, and substituting 4-benzoylbenzoic acid (31.3 mg, 0.138 mmol) for 2,2-bis-(hydroxy-methyl)-propionic acid. Chromatographic elution was with 2%
methanol in CH2C12. The title compound was obtained and dried in vacuo, ovemight.
m.p.: 110-135C
NMR: Consistent vvith structure HPLC: >94% pure MS: M + H 639 (FAB) Analysis calculated for C3gH42N2O~S0.40C6H14 C, 72.C6; H, 7.13; N, 4.16 Found: C, 72.CI; H, 7.14; N, 4.18 .

D02 ~
- OH,p )H
`N~N~1~0~<
1 '-(bicyclo[2.2. 1 ]hept-5-en-2-ylsulfonyl)spiro[ I H-indane- 1,4'-piperidinel The procedure of Example 42 was carried out using 94.5 mg, 0.22 mmol of (IS)-(((7,7-dimethyl-(2-endo-amino-methyl-2-exo-hydroxy)-bicicylo-(2.2. 1 )-hept- I -yl)-methyl)-sulfonyl)spiro( I H-indene-1,4'-piperidine), 49.8 mg, 0.26 mmol of EDC, 35.2 mg, 0.26 mmol of HBT, 90 mL, 0.66 mmol of triethylamine, and substituting t-boc-L-20 serine(bzl) (76.8 mg, 0.26 mmol) for 2,2-bis-(hydroxymethyl)-propionic acid. Chromatographic elution was with 98/2 of CH2C12/methanol/ammonium hydroxide. The title compound was obtained as a white solid from ether and dried in vacuo overnight.
25 m.p. 75-95C
NMR: Consistent with structure HPLC: >9~% pure MS: M + H 639 (FAB) Analysis ~ t~d for C3gHs3N3O7S
3~ C, 66.16; H, 7.55; N, 5.94 Found: C, 66.04; H, 7.68; N, 5.84 WO 95/14025 PCr/US94/13483 2i7~65`a OHII

H
[lS-[l.alpha., 2 Beta., 2(R*), 4. beta.]]-2-amino-N[[1-[[(2,3-dihydro spiro[ 1 H-indene- I ,4'-piperidin] -yl)sulfonyl]methyl] -2-hydroxy-7,7 -dimethylbicyclor2.2.1 lhept-2-lylmethvll-3-hydroxy-propanamide The product of Example 44 (120 mg, 0.17 mmol) was dissolved in 4% formic acid in methanol. Palladium hydroxide catalyst (25 mg) was added to the solution and it was placed on a PARR
apparatus under sa~ p.s.i. of hydrogen, overnight. The reaction mixlure 20 was filtered over solka floc and the filtrate was concentrated to a clear oil. This clear oil ~as dissolved in 2 mL of ethyl acetate and chilled to 0C. At this point, ~ mL of prechilled sat'd HCI/ethyl acetate solution was added dropwise. The reaction mixture was allowed to stir for I
hour and then concentrated to dryness. The resulting residue was 25 partitioned between ethyl acetate and saturated sodium bicarbonate, and after extracting 3 times with ethyl acetate, the organics were combined and dried over sodium sulfate. The organic layer was filtered, the filtrate was concen~:rated, and the resulting residue was dissolved in CH2C12. This was then poured onto a silica gel column and eluted witl 30 95/5/0 5 of CH2C12/methanol/ammonium hydroxide. The product fractions were combined and evaporated to dryness. The title compound was obtained as a white .solid from ether and dried in vacuo overnight.

WO 9~11402~ PCr~US94/13483 211465~

m.p.: 70-1 lO~C
NMR: Consistent with structure HPLC: >93% pure - MS: M +H 520.3 (FAB) Analysis calculated for C27H41N3OsS0.25 C4Hlo C, 61.31; H, 8.20; N, 7.67 Found: C, 61.35; H, ~S.11; N, 7.58 SOz - ~
H

3-[[[[2-hydroxy-7,7-dimethyl-1-[(spiro[lH-indene-1,4'-piperidin]-1'-ylsulfonyl)methyl]bicyclo[2.2.1]-hept-2-yl]methyl]amino]carbonyl]-bicyclor2.2. 1 lhept-5-ene-2-carboxylic acid ( 1 S)-(((7 ,7-dimethyl-(2-endo-aminomethyl-2-exo-hydroxy)-bicicylo-(2.2. 1 )~hept- I -yl)-methyl)sulfonyl)spiro( I H-indene-1,4'-piperidine) (98.5 mg, 0.23 mmol) was combined with bicyclo (2.2.1)-5-heptene-2,3-dicarboxylic anhydride (43 mg, 0.26 mmol) in 5 mL of THF. Triethylamine (50 mL, 0.36 mmol) was added to the reaction dropwise to adjust the pH to 9. The reaction mixture was stirred overnight and then concentr~ted to dryness. The resulting re.sidue was dissolved in CH2cl2~ poured onto a silica gel column, and eluted with 971310.3 of chloroform/methanol/acetic acid. The product fractions were combined and concentrated to dryness. The title compound was obtained as a white solid from ether and dried in vacuo overnight.
m.p.: 128-16~C
5 NMR: Consistent ~vith structure HPLC: >95% pure MS: M + H 595 (FAB) Analysis r~lc~ tt~d for C33H42N206SO.10 CH2C12 C, 64.74- H, 7.12- N, 4.56 Found: C, 64.76, H, 7.07, N, 4.66 NH~
o ( 1 S)- I '-(((2-endo-cyanomethyl-7,7-dimethyl-exo-2-hydroxybicyclo-25 (2.2.1)-hept-1-yl)methyl)sulfonvl)-spiro(lH-indene-1.4'-piperidine) Lithium bis(trimethylsilyl)amide (21.3 ml of a 1.0M THF
solution, 21.3 mmol) was cooled to -7~C under N2, treated with acetonitrile (1.07 ml, 20.4 mmol) and stirred 15 mmutes. A THF
solution (30 mL) oi` ( I S)- I '-(((7,7-dimethyl-2-oxobicyclo( 2.1 )-hept- I -30 yl)methyl)sulfonyl)spiro(lH-indene-l .4'-piperidine) (40 gm. 10 mmol) was added dropwise and stirred 10 minutes after addition was complete.
6N HCI (2.R ml) w~ls added to the cold reaction all at once and the mixture was allowed to warm to 25C.
The mixture was diluted with H2O (25 ml) and extracted with EtOAc (3 x 100 ml). The or~anic layers were combined~ washed WO 9~/14025 Pcr/uss4ll34s3 21 7~5~

with H20 (25 ml) and brine (25 ml), dried over Na2SO4, filtered and concentrated to dryness in vacuo. Trituration with ether gave the title compound (4.17 gm, 93% yield) as a crystalline solid (m.p. 199-202C).
TLC: Rf = 0.69, Silica GF (7% Et2O in CH2cl2) PMR: Consistent with structure ( I S)- 1 '-(((2-endo-aminoethyl-7,7 -dimethyl-2-exo-hydroxybicyclo-(2.2.1 )-hept- I -yl)methyl)~ulfonyl)spiro( I H-indene- I ~4'-piperidine) IJnder a blanket of nitrogen, Lithium Alllminllm Hydride (8.0 ml, 8.0 mmol, of a 1.0 M THF solution) was cooled to 0C, treated dropwise with a THF solutioP (37 ml) of (IS)-1'-(((2-endo-cyano-methyl-7 ,7-dimethyl-2-hydroxybicyclo-(2.2. 1 )-hept- I -yl)methyl)-sulfonyl)spiro(lH-indene-1,4'-piperidine) (4.4 gm, 10.0 mmol) and the reaction stirred 10 minutes. With rapid stirring, the reaction was treated with H20 (0.37 ml), 20% NaOH (aq) (0.40 ml) and H2O (1.46 ml) followed by extraction with EtOAc (3 x 150 ml). The organic layers were combined, washed with H2O (25 ml) and brine (25 ml), dried over Na2SO4, filtered and cu~ ld~d to dryness in vacuo.
Flash chromatography on silica gel (90/10/1/1 of CH2C12/MeOH/H20/HOAc) provided product fractions which were pooled and concentrated to dryness. The residue was treated with sat'd Na2CO3 (aq) and extracted with EtOAc (3 x 100 ml). The orgarlic layers were combined, washed with H2O (25 ml) and brine (25 ml), 25 dried over Na2SO4, filtered and coricentrated to dryness to give the title compound (2.55 gm, 57% yield) as a white foam/solid.
TLC: Rf = 0.27 Silica GF (90/1011/1 of CH2cl2lMeoHlH2olHoAc) PMR: Consistent with structure (lS)-I '-(((2-endo-aminoethyl-7,7-dimethyl-2-exo-hydroxybicyclo-(1.2. 1 )-hept- I -yl)methyl)sulfonyl)-spiro( I H-indene-1,4'-piperidine) (6û mg. 0.135 mmol) was dissolved in CH2C12 (I ml), treated with 4-iodobenzoyl chloride (3~.0 mg, 0.135 mmol) and Et3N
_ . . . ..

2l7~sa to adjust the pH to 9.5. The reaction was stirred 30 minutes at 25C and flash chromatograplhed on silica gel (8% Et2O in CH2C12) to give the title compound (45 mg, 50% yield) as a white foam from Et2O (m.p.
98-138C, shrinks).

TLC: Rf = 0.35 Silica CF (10% Et20 in CH2cl2) PMR: Consistent ~lith structure HPLC: 95.8% pure M.S.: (FAB) M + H + Thioglycerol = 783 o Analysis calculated for C32H3gIN2O4s0 30c4Hloo;o 35 H2O
C, 56 70; H, 6.12; N, 3.98 Found: C, 56 73; H, 6.11; N, 4.01 ~`0~
= OH A
NH~

(lS-exo-N-[2-[2-hy~roxy-7,7-dimethyl-1 -[(spiro[lH-indene-l ,4'-piperidin] I '-ylsulfonyl)methyl]bicyclo[2.2. 1 ]hept-2-yl]ethyl]-cyclo-propanecarboxamide The title compound was prepared according to the 30 procedure of Exarnple 47 except that cyclopropylcarbonyl chloride (12.4 ml, 0.135 mmol) was s~-h~tih-t~-d for 4-iodobenzoyl chloride.
Flash chromatography of the reaction mixhure on silica gel (15% Et2O in CH2C12) gave the title compound (36.6 mg, 53% yield) as a white foam from Ether (m.p. 93-105C shrinl; + foam).
t WO 95114025 PCrlUS94113483 21 74~50 TLC: Rf= 0.22 Silica GF (15% Et2O in CH2cl2) PMR: Consistent with Structure HPLC: 96.8%
- M.S.: (FAB) M + H + Thioglycerol = 621 Analysis calculated for Calc'd for C2gH40N2O4S0.20C7H1oO
C, 67.84; H, 8.03; N, 5.31 Found: C, 67.69; H, 8.06; N, 5.06 52~ OH
N~

(lS-exo)-N-[2-[2-hydroxy-7,7-dimethyl-1-[(spiro[lH-indene-1 ,4'-piperidin[- I '-ylsulfonyl)Methyl]bicyclo[2.2. 1 ]hept-2-yl]ethyl] -2,2-dimethyl -propanamide The title compound was prepared according to the 25 procedure of Example 47 except that trimethyl acetyl chloride (16.6 ml7 0.135 mmol) was s~lb~titllt~d for 4-iodobenzoyl chloride.
Flash chromatography of the reaction on silica gel (13%
Et2O in CH2C12) gave the title compound (32.8 mg, 46% yield) as a white foam from Et2O (m.p. 78-104C, shrink + foam).
TLC: Rf = 0.2~ Silica GF (15% Et2O in CH2cl2) PMR: Consistent with structure HPLC: 97.6%
M.S.: (FAB) M + H + Thioglycerol = 637 Analysi~ calculated for C30H44N2O4S0.20C4HIoO0.30 H2O =

2~ 74~

C, 67.38; H, 8.56; N, 5.10 Found: C, 67.37; H, 8.52; N, 5.02 OH ~ ~NOz HN~

(IS-exo)-N-[2-[2-hydroxy-7,7-dimethyl-1-[(spiro[lH-indene-1,4'-piperidin] - I '-ylsulfonyl)methyl]bicyclo[2.2. 1 ]hept-2-yl]ethyl] 4-methoxy-benzamide ~._ The title compound was prepared according to the 20 procedure of Exam]?le 47 except that 4-nitrobenzoyl chloride (25.1 mg, 0.135 mmol) wa~ sllhstitl-t~d for4-iodobenzoyl chloride.
Flash chromatography of the reaction on silica gel (10%
Et2O in CH2C12) ga~e the title compound (41.6 mg, 52% yield) as a white foam from Ether (m.p. 11 8-33C, shrink + foam).
TLC: Rf= 0.26 Silica GF (10% Et2O in CH2C12).
PMR: Consistent with structure HPLC: 97.1%
M.S.: (FAB) M + ~1 + Thioglycerol = 702.
30 Analysis ~ t~d for C32H3gN3O6S0.30C4H10O0.20 H20 C, 64.35; H, 6.90; N, 6.7 Found: C, 64.32; H, 6.9~; N, 6.63 wo 9~/1402~ Pcrlus94ll3483 2~ 74~5~

HN~
(IS-exo)-N-[2-[2-hydroxy-7,7-dimethyl-1-[(spiro[lH-indene-1 ,4'-piperidin]-l'-ylsulfonyl)methyl]ethyl]bicyclo[2.2.1]hept-2-yl]ethyl[-4-5 methoxy-benzamide The title compound was prepared according to the procedure of Example 47 except that p-anisoyl chloride (18.3 ml, 0.135 mmol) was s-lhstitllt.od for 4-iodobenzoyl chloride.
Flash chromatography of the reaction on silica gel (15 %
20 Et2O in CH2C12) gave the title compound (36 mg, 46% yield) as a white solid from Et20 (m.p. 98-121~C, shrink).
TLC: Rf= 0.26 Silica GF (15% Et2O in CH2C12) PMR: Consistent with structure HPLC: 955%
M.S.: M + H + Thioglycerol = 687 Analysis calculated for C33H42N20sS0.25 C4HloO
C, 68.36; H, 7.51; N, 4.69 Found: C, 68.14; H, 7.67; N, 4.55 21 746~0 HN~ NH~,~
O
(lS-exo)-1'-[[[2-[2-[[[(1 ,1 -dimethylethyl)amino]-carbonyl]amino]-ethyl] -2-hydroxy-7,7-dimethylbicyclo-[2.2. 1 ]hept- I -yl]methyl] -sulfonyll-spirorlH-indene-1~4'-piperidinel (IS)-I '-[[[endo-aminoethyl-7,7-dimethyl-exo-hydroxy-bicyclo(2.2.1)-hept l-yl]methyl]sulfonyl]spiro(lH-indene-1,4'-piperidine) (60 mg, 0.135 mmol) was dissolved in THF (2 ml), treated with t-butyl isocyanate (15.4 ml, 0.135 mmol) and stirred 30 minutes at After removal of the solvent in vacuQ, the residue was flash chromatographed on silica gel (25% Et2O in CH2C12 to give the title compound (27.9 m~, 38% yield) as a white solid from Et2O (m.p. 97-119C, shlink and foam).
TLC: Rf= 0.21 Silica GF (20% Et2O in CH2C12)PMR: Consistent with structure HPLC: 95 1%
M.S.: (FAB) M + 11 + Thioglycerol = 652 Analysis calculated for C30H4sN3O4S0.25C4H10O0.30 H2O
C, 65.58; H, 8.54; N, 7.40 Found: C, 65.70; H, 8.74; N, 7.05 WO 95/14025 PCrrUS94rl3483 5 ~
~; ~
~`S~O
O ~No~~
To a stirred solution of 10 ml of dry N,N-dirnethyl-formamide Gonts~inin~ endo(lS)-1'-(((2-amino-7,7-dimethyl-bicyclo(2.2.1) hept-1-yl)methyl)sulfonyl)-spiro(lH-indane-1,4'-piperidine) (670 mg, 1.66 mmole) and Na-tert-butyloxycarbonyl-5,5-dimethyl-L-thiazolidine-4-carboxylic acid (500 mg, 1.91 mmole) was added 932 mg (2.11 mmole) of be~ ol-l-yloxytris (dimethyl-amino)phosphonil~m hexafluorophosphate. The pH of the reaction mixture was adjusted to 8.~ with diisopropylethylamine and the reaction 2s mixture was stirred at 23C overnight. The reaction mixture was filtered and concentrated under reduced pressure. The residue was dissolved in ethyl acetate and this solution was washed in succession with saturated sodium bicarbonate solution (3 X 25 ml) and brine. The organic phase was dried (sodium sulfate) and concentrated to yield a 3 brown solid which was purified by chromatography on silica gel (chloroform-methanol-concentrated ammonium hydroxide elution, 98:2:0.2 v/v) to yield a white solid. This material was dissolved in 10 ml of chloroform. The solution was cooled in an icebath and treated dropwise with a solution of 5 ml of chloroforrn containing 2.2 wo 9~/14025 PCr~S94/13483 211~6~0 equivalents of m-chloroperoxy-benzoic acid. After addition was complete the ice bath was removed and stirring was continued for 24 hours. The reactioll mixture was diluted to 100 ml with choroform and was washed with lhl sodium hydroxide solution (2 X 30 ml) and brine.
5 The organic phase was dried (sodium sulfate) and concentrated under reduced pressure. The crude product was purified by chromatography on silica gel (chloroform-methanol-concentrated ammonium hydroxide elution, 98:2:0.2 v/~) to yield the title compound:
o NMR: Consistent with structure and verifies presence of solvent;
HPLC: >94% pure at 214 nM;
FAB MS: 678 (M+ + H);
Elem Analysis calculated for C34HslN3O7S2-0.8CHC13 ~, 54.04; H, 6.75; N, 5.43 Found: ~, 54.11; H, 6.64; N, 5.48 ~2 2s 1 ~ ~
O~NH2 HN ~ ~N
COOCH

WO 95/14025 PCr/US94113483 Endo(lS)-1 '-(((2-amino-7,7-dimethylbicyclo(2.2.1)hept-1-yl)methyl)sulfonyl)spiro(l H-indane- I ,4'-piperidine)(660 mg, 2.05 mmole) and (S)-3-[(tert-butyloxycarbonyl)amino]-2-oxo-1-pyrrolidine-(R)-2-methylcarboxysuccinic acid (826.5 mg, 2.05 mmole) were combined with 392 mg (2.05 mmole) of 1-ethyl-3-(3-dimethylamino-propyl) carbodiimide hydrochloride and 275 mg (2.05 mmole) of 1-hydroxybenzotriazole hydrate in 20 ml of dry methylene chloride at room ~ eldLul~ under nitrogen. The pH of the reaction mixture was adjusted to 8.5 with triethylamine and the resulting solution was stirred o for 12 hours. The reaction mixture was diluted with methylene chloride (150 ml) and the resulting solution was washed with saturated sodium bicarbonate solution (2 X 40 ml), 10% citric acid solution (2 X
40 ml), and brine, then dried (m~gn~ m sulfate) and concentrated to give the crude product. The analytically pure material was obtained via preparative HPLC chromatography employing a Vydac C-18 column (4.5 X 150 mm, water-acetonitrile-1% trifluoroacetic acid 45 minute gradient). The homogeneous fractions c--nt~inin~ product were pooled and cullc~ d. The residue was dissolved in methylene chloride containing trifluoroacetic acid (50%)at room temperature. After 2 hours, the volatiles were removed under reduced pressure to yield the title compound as a trifluoroacetate salt:
NMR: Consistent with structure and confirms presence of solvent;
HPLC: > 97% pure at 214 nm;
FABMs:6l5(M++H);
Elem Analysis calculated for C34H47F3N4OgS-0.3 H20-0.8TFA:
C, 51.59; H, 5.88; N, 6.74.
Found: C, 51.59; H, 5.89; N, 6.81.

WO 95/~4025 PCTIIIS94/13483 217~0 o~,7 HN--~--I`NH~.~O~
o Endo-( I S)-l '-(((2-amino-7,7-dimethylbicyclo(2.2. 1 ) hept-20 1-yl)methyl)sulfon~ll)spiro(lH-indane-1,4'-piperidine)(216 mg, 0.53 mmole) and Na-terl-butyloxycarbonyl-D~L-(3-thienyl)alanine (160 mg, 0.59 mmole) were combined with 113 mg (0.59 mmole) of 1-ethyl-3-(3-dimethylaminop~opyl) carbodiimide hydrochloride and 79 mg (0.59 mmole) of l-h~dlu~y~ lz~ vle in 6 ml of dry methylene chloride at 25 room te,l.l,~.a~u~ mder nitrogen. The pH of the reaction mixture was adjusted to 9 with triethylamine and the resulting solution was stirred for 12 hours. An additional 20 mg of tert-butyloxycarbonyl-D,L-(3-thienyl)alanine, 11 mg of 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride were added and stirring was continued for 30 6 hours more keeping the pH of the medium between 8 and 9. The reaction mixture wa.~ diluted with methylene chloride (150 ml) and the resulting solution was washed with saturated sodium bicarbonate solution (2 X 40 ml), 10% citric acid solution (2 X 40 ml), and brine, then dried (magnesium sulfate) and concentrated to give the crude product. The analytically pure material was obtained via flash column 21 7~50 chromatography on silica gel (chloroform-methanol-concentrated ammonium hydroxide elution, 95:5:0.5 v/v):
NMR: Consistent with structure and confirms presence of solvent;
5 HPLC: > 99% pure at 214 nm;
FAB MS: 656 (M+ + H);
Elem Analysis calc'd for C3sH49N3sS2 -5 H2O:
C, 63.22; H, 7.58; N, 6.32.
Found: C, 63.26; H, 7.54; N, 6.10.

15 ~J
S

CO2CH2CH~
Endo( I S)- I '-(((2-amino-7 ,7-dimethylbicyclo(2.2. 1 ) hept- I -30 yl)methyl)sulfonyl)spiro(lH-indane-1,4'-piperidine)(67 mg, 0.156 mmole) and ~I-ethyl-carboxymethyl-5,5-dimethyl-L-thiazolidine-4-carboxylic acid (57 mg, 0.23 mmole) were combined with 44 mg (0.23 mmole) of l-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride and 31 mg (0.23 mmole) of l-hydroxybenzotriazole wo 95/1402s PCrlUS94113483 .

hydrate in 5 ml of dry methylene chloride at room temperature under nitrogen. The pH of the reaction mixture was adjusted to 8.5 with triethylamine and the resulting solution was stirred for 3 hours. The reaction mi~ture was diluted with methylene chloride (50 ml) and the 5 resulting solution was washed with saturated sodium bicarbonate solution (2 X 40 ml), 10% citric acid solution (2 X 40 ml), and brine, then dried (m~gJn~ m sulfate) and concentrated to give the crude product. The analytically pure material was obtained via preparative thick layer chromatography (2 X 20 X 20 mm, pre-coated SiO2 plates, o hexane-ethyl acetate elution, 2:1 v/v). The analytical product was isolated in homogeneous form as a solid:
NMR: Consistent with structure and confirms presence of solvent;
FAB MS: 660 (M+ + H);
Elem. Analysis calc'd for C34H47N3O6S2-0.85 H2O-O.lEtOAc:
C, 60.40; H, 7.59; N, 6.14.
Found: C, 60.37; H, 7.36; N, 6.13.
lEXAMPLE 57 `T' H~
.

-WO9~/14025 21 i~b~ PcTruss4rl34s3 To a stirred solution of 50 ml of dry N,N-dimethyl-formamide containing endo(lS)-1'-(((2-amino-7,7-dimethyl-- bicyclo(2.2. 1 ) hept- I -yl)methyl) sulfonyl)spiro(1 H-indane- 1,4'-5 piperidine) (4.0g, 9.9 mmole) and Na-tert-butyloxycarbonyl-L-methioninesulfone (2.84 g, 11.9 mmole) was added 5.65 g (12.87 mmole) of benzotriazol-l-yloxytris(dimethylamino) phosphonium hexafluorophosphate. The pH of the reaction mixture was adjusted to 8.5 with diisopropylethylamine and the reaction mixture was stirred at o 23C for two hours. The reaction mixture was filtered and concentrated under reduced pressure to give a solid. This material was dissolved in ethyl acetate and this solution was washed in succession with saturated sodium bicarbonate solution (3 X 25 ml), 10% citric acid solution (3 X 25 ml) and brine. The organic phase was dried (sodium 5 sulfate) and concentrated to yield 2-tert-butyloxycarbonylamino-N-[I-[[(2,3-dihydrospiro[lH-indene-1,4'-piperidin]-1'-yl)sulfonyl]methyl]-7,7-dimethylbicyclo[2.2. 1 ]hept-2-yl]-4-(methylsulfonyl)-k~ n~mi~
which was purified by chromatography on silica gel (chloroforrn-methanol-concentrated ammonium hydroxide elution, 95:5:0.5 v/v) to 20 yield a solid. This material was dissolved irl 50 ml of ethyl acetate and the resulting solution was cooled to 0C and treated with a continuous stream of hydrogen chloride gas for 15 minutes. The ice bath was removed, the reaction vessel was capped, and stirring was continued for 45 minutes more at room ~ lp~ldtul~. The solvent and excess 25 hydrogen chloride were removed under reduced pressure to give 2-amino-N-[ I -[[(2,3-dihydrospiro[ I H-indene- I ,4'-piperidin]- I '-yl)-sulfonyl]methyl]-7,7-dimethyl-bicyclo[2.2. 1 ]hept-2-yl]-4-(methyl-sulfonyl)butanamide hydrochloride as an off-white solid (6.5 g).

wo 9S/14025 PCrlUs94ll34S3 2~

~o~ i \ 0~,0 HN _~/
N
o 2-arnillo-N-[1-[~(2,3-dihydrospiro[lH-indene-1,4'-piperidin]-1'-yl)sulfonyl]methyl]-7,7-dimethylbicyclo[2.2.1]hept-2-yl]-4-(methylsulfonyl)-butanamide hydrochloride was chromatographed on 20 silica gel (chloroform-methanol-concentrated ammonium hydroxide elution, 90:10:1 v/~/) to afford a white solid which was dissolved in 15 ml of methanol cf-n~:linin~ 1% acetic acid. To this reaction mixture wa~
added 3 ml of aqueous formaldehyde solution and 1.56 g (24.8 mmole) of sodium cyanoborohydride. The resulting reaction mixture was 25 stirred at room ~ ueldLu~ overnight. The reaction mixture was concentrated in vacuo and the residue was partitioned between ethyl acetate and saturated sodium bicarbonate solution. The phases were separated and the organic phase was washed with brine, dried (sodium sulfate), and conc~ntrated under reduced pressure. The crude product 3~ was purified by ~ ul~latography on silica gel (chloroform-methanol-concentrated ammonium hydroxide elution, 95:5:0.5 v/v) to yield 2-dimethylamino-N-[1-[1(2,3-dihydrospiro[lH-indene-1,4'-piperidin]-1'-yl)sulfonyl]methyl]-7,7-dimethylbicyclo[2.2. 1 ]hept-2-yl] -4-(methyl-sulfonyl)bllt~n~mi~

NMR: Consistent with structure and verifies presence of solvent;
HPLC: >98% pure at 214 nM;
FAB MS: 594(M+ + H);
Elem. Analysis calc'd for C30H47N3OsS2-0.1CHC13:
C, 59.67; H, 7.84; N, 6.94.
Found: C, 59.90; H, 7.24; N, 6.93.
~XAMPLE 58 ~X~
N
0,~
OH
HN~ ~N b,O~¦

To a stirred solution of (IS)-1'-(((2-exo-hydroxy-2-endo-aminomethyl-7 ,7-dimethylbicyclo(2.2. 1 )hept- l -yl)methyl)sulfonyl)-spiro(lH-indene-1,4'-piperidine) (300 mg, 0.694 mmole) in 10 ml of dry, degassed N,N-dimethylformamide was added Na-tert-butyloxy-carbonyl-S-methyl-L-cysteine (212 mg, 0.832 mmole) and 398 mg (0.902 mmole) of benzotriazol-l-yloxy tris(dimethylamino)-phosphonium hexafluorophosphate (BOP) at room I~ UI~. The resulting reaction mixture was protected from moisture and the pH wa~
adjusted to 8-9 with diisopropylethylamine. After 24 hours all volatile wo 95/14025 Pcr/usg4/l3483 2 ~ a components were removed under reduced pressure and the residue was partitioned between ethyl acetate and sodium bicarbonate solution. The phases were separated and the organic phase was washed with saturated sodium bicarbonate solution (3 X 40 ml) and brine, then dried 5 (m:l~nP~illm sulfate) and concentrated. The crude product was chromatographed on silica gel (hexane-ethyl acetate elution, 3:1 v/v) to give the title compound:
NMR: Consistent with structure and verifies presence of solvent;
o HPLC: >98% pure at 214 nM;
FAB MS: 650 (M+ ~ H);
Elem. Analysis calc'd for C33H50N3o6s~:
C, 61.07; H, 7.71; N, 6.47.
Found: C, 61.24; H, 7.98; N, 6.13.

~
2s lsJz,~
O
HN ~NH
~<
O HN~, o To a stirred solution of (lS)-1'-(((2-exo-hydro~y-2-endo-aminomethyl-7,7-dimethylbicyclo(2.~.1 )hept- I -yl)methyl)sulfonyl)-spiro(lH-indene-1,1'-piperidine) (125 mg~ 0.311 mmole) in 7 ml of WO 9511402~ PCr/lJS94/13483 21 74~5~

dry, degassed N,N-dimethylformamide was added S-hydantoinacetic acid (59 mg, 0.372 mmole) and 178 mg (0.404 mmole) of benzotriazol-1-yloxy tris(dimethylamino)pho~ ulliu ll hexafluorophosphate (BOP) at room ~ e~dlu~. The resulting reaction mixture was protected 5 from moisture and the pH was adjusted to 8-9 with diisopropyl-ethylamine. After 24 hours all volatile components were removed under reduced pressure and the residue was partitioned between ethyl acetate (100 ml) and sodium bicarbonate solution. The phases were separated and the organic phase was washed with saturated sodium bicarbonate solution (3 X 40 m]) and brine, then dried (magnesium sulfate) and concentrated. The crude product was initially flash chromatographed on silica gel (chloroform-methanol elution, 96:4, v/v) to give semi-pure material from which the title compound was obtained as a white solid after preparative thick layer chromatography on silica 15 gel (chloroforrn-methanol-concentrated :~mmonillm hydroxide elution, 96:4:0.4, v/v) and trituration with ether-petroleum ether:
NMR: Consistent with structure and verifies presence of solvent;
FAB MS: 543 (M+ + H);
Elem. Analysis calculated for C28H48N4O5S
C, 61.96; H, 7.00; N, 10.32 Found: C, 61.70; H, 7.36; N, 10.09 wo 95114025 Pcrlusg4ll3483 2 1 74~0 EX~MPLE 60 10 \ ~
¦ H
HN
15 o~ N~/~
N

0~0~
(1S)-1~-(((7,7-dimethyl-2-endo-(4-nitrophenyloxycarbonylamino)-bicyclo-(2.2.1)-hept-1 -yl)-methyl)-sulfonyl)spiro(lH-indan-l ,4'-~iperidine (lS)-1'-(((7,7-dimethyl-(2-~n~ mino)-bicyclo(2.2.1)-hept-I-yl)-methyl)sulfonyl)spiro(lH-indene-1,4-piperdine)[3.47mmol] and 4-Nitrophenyl chloro~ormate [3.64mrnol] were combined in THF. The reaction mixture W~IS treated with triethylamine [4.54 mmol] and allowed to stir for 3 hours. The reaction mixture was concentrated to dryness and the reslulting residue was purified by a silica gel column, while eluting with l % ethylacetate in methylene chloride. The product fractions were combined and concentrated to dryness in vacuo. (IS)-I'-(((7,7-dimethyl-(4-~itrophenyloxycarbonyl-2-endo~minr )-bicyclo-(2.21 )-hept- I -yl)-miethy~)-sulfonyl)spiro( I H-indene- 1 ,4'-piperidine was obtained as a white solid from ether.
(lS)-I '-(((7,7-dimethyl-2-endo-(4-nitro-phenyloxy-carbonylamino)-bicicylo-(2.2.1)-hept-1-yl)-methyl)sulfonyl)spiro(ll~-wo 95/14025 Pcr/usg4/l3483 21 74~

indene-1,4'-piperdine)-[0.278mmol] and (benzyloxycarbonyl) piperazic acid [0.334mmol] were combined in DMF. The reaction mixture was treated with triethylamine [0.401mmol] and allowed to stir for 2 hours.
The reaction mixture was concentrated to dryness and the resulting 5 residue was dissolved in CH2cl2. This solution was placed on a silica gel column and eluted with 5% methanol in CH2cl2 and then with 96/4/0.4 of CH2C12/methanol/acetic acid. The product fractions were combined and evaporated to dryness. The title compoumd was obtained as a white solid from ether and was dried in vacuo overnight.
m p.: 9o-l2ol~c NMR: Consistent with structure HPLC: >98% Pure MS: M-->H+=693.6 Analysis calculated for C37H4gN4O7S-0.20mol C4HloO-0 25mol H20 C, 63.74; H, 7.15; N, 7.87 Found: C, 63.78; H, 7.08; N, 7.81 HN
~ N ~O

~VO 95/14025 PCr/US94113483 The procedure of Example 60. second paragraph was carried out using (lS)-1'-(((7,7-dimethyl-2-endo-(4-nitrophenyloxy-carbonylarnino)-bicyclo-(2.2. 1 )-hept- 1 -yl)-methyl)sulfonyl)spiro( 1 H-indan- 1 ,4'-piperidine)[0. 1 93rnmol], triethylamine [0.29mmol~, and 5 ~ "l;"g 3-(t-butoxycarbonylarnino-methyl)piperidine [0.212mrnol]
for (benzyloxycarbonyl) piperazic acid. Chromatographic elution was with 5% ether in CH2C12, 10% ether in CH2cl27 and then 1% methanol in CH2C12. The title compound was obtained from ether and dried in vacuo, overnight.
o m p 95-105C
NMR: Consistent with structure HPLC: >97% Pure MS: M+H+=643.4 Analysis calculated for C3sHs4N4OsS-0 20 H2O
C, 65~2; H, 8.48; N, 8.67 Found: C, 64.~8; H, 8.43; N, 8.86 WO 95J140~5 PCr/US94/13483 2174~50 Endo-( I S)- I '-(((2-(L-4(tert,butoxycarbonylamino)glutaramyl)amino-7,7-dimethylbicyclo(2.2. 1 )hept- I -yl)-methyl)-sulfonyl)spiro-( I H)-indan- I .4'.piperidine N r7`
~
H '~NH2 In an oven dried flask (50ml) under nitrogen was dissolved endo-(lS)-1'-(((2-amino-7,7-dimethyl-bicyclo(2.2.1)hept-1 -yl)-methyl)-sulfonyl)spiro-(lH)-indan-1,4'-piperidine (50mg, 0.125mmol), L-4(tert-butoxycarbonyl)glutaramic acid (34mg, 0.14mmol), 1-ethyl-3-(3-dimethyla..li-.op.vl yl)carbodiimide hydrochloride (30mg, 0.15mmol), and l-hydroxybenz-triazole hydrate (20mg 0.15mmol) in dimethyl formamide (Iml). Triethylamine (50ml) was added and a white solid separated. After stirring for I hour, the solvent was removed under reduced pressure. The residue was dissolved in ether:methylene chloride (3:1). The cloudy solution was washed with sodium bicarbonate (sat., aqueous), water, potassium hydrogen sulfate (10%, aqueous) and brine. After drying over sodium sulfate, the organic material was filtered and concentrated. Silica "flash" chromatography using methylene chloride methanol (9:1) gave the product which was isolated as an oil upon evaporation of the solvents. Hexane:ether WO 9~i/140~i PCIIUS94/13483 2 l ~ 4 63 addition and remov~ll under reduced pressure produced the title compound as a white foam.
HPLC: >99%
MS: M+H+=63 1.3 NMR consictpnt with structure Analysis calcula~ed for C33HsoN4O6S-0.5 DMF-0.5 H20 C, 61.26; H, 8.12; N, 9.32 Found: C, 6127; H, 8.05; N, 9.31 Endo-( l S)- I '-(((2-(4 (imidazole-2-ethylacetyl)amino-7,7 -dimethyl-bicyclo(2.2. 1 )hept- l -yl)-methyl)spiro-( 1 H)-indan-( I ,4'),piperdine 1~ hydrochloride N
2s H ,CH3 CH -Step 1: Endo-( I S)- I '-(((2-4(3-methylbenzyloxy)imida~ole)-2-[ethyl acetyl)amino-7,7-dimethylbicyclo- (2.2.1 )hept- I -vl)-methyl)-sulfonyl)spiro( I H)-indan-( I .4')-pi~eridine Into an over dried flask (50ml) under nitrogen was placed dimethylformamide (4ml). Endo( I S)- I '-((2-amino-7 ,7-dimethyl-WO 95/140~5 PCT/US94/13483 21 ~46~

bicyclo(2.2.1)hept-1-yl)-methyl)sulfonyl)spiro(lH)-indan-(1,4')-piperidine (300mg,0.74mmol), 4(3-methylbenzyloxy)imidazole-2-ethylacetic acid (219mg, 0.8mmol), 1-ethyl-3-methyl-(3-dimethyl-aminopropyl)carbodiimidehydrochloride-(153mg, 0.8mmol) and 1-hydroxybenztriazolehydrate-(108mg, 0.83mmol) were added and the mixture was stirred until solution was achieved. Triethylamine (400ml) was added to ma~e the solution basic (pH 9) and a solid separated.
After stirring ovemight room l~,l,pe,d~ul~, t'ne solvent was removed under reduced pressure. The resulting ~um was dissolved in o methyenechloride:ether (1:3) and the cloudy solution was extracted with sodium bicarboante (sat., aqueous) and brine. After drying the organic layers over sodium sulfate, the solution was filtered and was concentrated. The product was purified by silica gel "flash"
chromatography using methylene chloride, methanol, ammonium hydroxide (9:1:1) as solvents. The product fractions were concentrated to give the title compound as white foam.
HPLC: >95% (42% + 53%) MS: M+H+=659.3 NMR (CDC13) cnn~ist.-nt with structure Arlalysis Ç~ tPd for C3gHsoN404S-0.65H20 C,68.05; H,7.71; N,8.36 Found: C,68.03; H,7.87; N,8.60 2S Nitrogen was bubbled into a solution of Endo(lS)-1'-(((2-4-(3-methylbenzyloxyimidazole)-2-ethylacetylamino-7 ,7-dimethyl -bicyclo(2.2.1)hept-I-yl)-methyl)-sulfonyl)spiro-(1H)-indan-(lm4')piperidine (50mg, 0.076mmol) in absolute ethanol (20ml)-acetic acid (Sml). After 5 minutes, 10%Pd/C(SOmg) was added and the mixture was hydrogentated at 55psi ovemight. The catalyst was - removed by filtration and the solvents were concentrated to dryness under reduced pressure. The residue was purified by silica gel "flash"
chromatography using methylenechloride, methanol,ammonium hydroxide (9:1:1) as solvent. The product fractions were collected and concentrated. The gum was redissolved in methylene chloride and wa.s 21 74~

filtered through a sintered glass funnel. After the solution was concentrated, the re.~idue was dissolved in ethyl acetate (5ml). The solution was cooled to 0 and it was saturated with hydrogen chloride gas (bubbling 5 minutes). The solvent was removed under reduced 5 pressure and ether was added and was removed to give the title compound as a white solid.
HPLC: >90% (42% + 45%) MS: M+H+=539.2 (freebase) o NMR (CDC13) consistent with structure Analysis calcula~ed for C30H42N4O3S-HC1 0.25 ether- 1 30H2O
C, 60.33; H, 7.86; N-9.08 Found: C, 60.33; H, 7.48; N-9.04 H H
HN~N~
o 3 SO2CH~;
To a stirred solution of the product of Example A (500 mg.
1.24 mrnol) in DMF (10 mL) was added the HCI salt of 2-(1-benzyloxy-methylimidazoly-5-yl)-4-methylsulfonylbutanoic acid (506 mg; 1.30 WO 95/14025 PCrllJS94113483 mmol), DIEA (0.70 mL; 4.0 mmol), and BOP (600 mg; 1.35 mmol).
After being stirred at ambient temperature for 14 h, the solvent was removed under reduced pressure. The residue was dissolved in EtOAc (75 mL) and washed with aqueous NaHC03 (3 x 50 mL). The or~anic 5 phase was dried (MgS04), filtered, and the solvent was removed under reduced pressure. The benzyloxymethyl protecting group was removed by dissolving the residue in a 4:1 solution of EtOH-HOAc (10 mL) and hydrogenating over palladium black (75 mg) under I atm of hydrogen for 24 h. The catalyst was removed by filtration through Celite and the o filtrate solvents were removed under reduced pressure. The residue was purified by pressurized silica gel column chromatography using a 50:50:7 by volume mixture of CHC13, EtOAc, and 4% NH40H-MeOH
as eluant. Two diastereomers were obtained and each was Iyophilized from water containing 1% TFA. The title compound is the lower Rf 15 isomer.
Higher Rf Isomer:
Analysis calculated for (C31H44N40sS2)-1.0 TFA-l.l H20 C, 52.45; H, 6.24; N, 7.37 20 Found: C, 52.69; H, 6.12; N, 7.32 TLC: Rf 0.33 (50:50:7 CHC13:EtOAc:4%NH40H-MeOH) HPLC (method A): retention time 8.96 min FABMS:m/z617(M++H) 25 IH NMR (300 MHz, CDC13): free base: d 7.70 (s, lH), 7.55 (br d, lH), 7.15-7.25 (m, 4H), 7.00 (s, IH), 4.35 (m, lH), 2.90 (s, 3H), 1.00 (s, 3H), 0.98 (s, 3H) Lower Rf Isomer:0 Analy~is calculated for (C31H44N405S2) 1 0 TFA-1-3 H20 C7 52.55; H 6.36; N 7.43 Found: C. 52.89; H 6.13; N 7.43 TLC Rf 0.28 (50:50:7 CHC13:EtOAc:4%NH40H-MeOH) HPLC (method A): retention time 9.15 min wo 9511402~ Pc~!r/Usg41l3483 ~ 1-/ 4 ~

FAB MS: m/z 617 (M+ + H) IH NM~ (300 MHz, CDC13): free base: d 7.72 (s, IH), 7.34 (br d, lH), 7.15-7.25 (m, 4H), ~.98 (s, IH), 4.35 (m, IH), 2.90 (s, 3H), 1.00 (s, 3H), 0.96 (s, 3H) S~
H

HN
~,NH2 O ~

To a 0C stirred solution of the product of E~ample A
(1.10 g; 2.74 mmol) in CHC13 (100 mL) was added DIEA (0.75 mL;
4.3 mmol) and benzyl chloroformate (0.51 g; 3.0 mmol). The solution was stirred at 0C for 1 h and then at ambient ~ eldlul~ for 14 h.
The reaction mixtule was concentrated under reduced pressure and the residue was dissolved in 25 mL of a solution of 10;1 MeOH-NH40H.
The mixture was concentrated under reduced pressure, the residue was dissolved in CHCI~ (100 mL) and washed with 5% aqueous HCI (2 x 50 mL) a~d aqueous NaHC03 (100 mL). The organic phase was dried (MgS04), filtered, and the solvent was removed urlder reduced pressure. The residue was purified by pressurized silica gel WO 9511402~ PCI~/US94/13483 21 74~5~

column chromatography using 1:4 EtOAc-hexanes as eluant. The benzyl urethane was obtained as a white foam.
TLC: Rf 0.40 (1:3 EtO~r h~Y~n~s) 5 HPLC (method A): retention time 12.25 min FAB MS: m/z 537 (M+ + H) To a 0C stirred solution of the benzyl urethane (1.25 g;
2.33 mmol) in DMF (20 mL) was added iodomethane (0.435 mL; 7.00 mmol) and sodium hydride (0.140 mg of a 60% dispersion in mineral oil; 3.50 mmol). The solution was stirrred at 0C for 1 h and then at ambient temperature for 18 h. The reaction mixture was treated with HOAc (1 mL) and the solvents were removed under reduced pressure.
The residue was dissolved in EtOAc (100 mL) and washed with aqueous NaHCO3 (2 x 50 mL). The organic phase was dried (MgSO4), filtered, 15 and the solvent was removed under reduced pressure. The residue was purified by plcs~u~ d silica gel column chromatography using 1:5 EtOAc-hexanes as eluant. The N-methyl benzyl urethane was obtained as a white foam.
TLC: Rf 0.34 (1:4 EtOAc:hexanes) 20 HPLC (method A): retention time 12.71 min FAB MS: m/z 551 (M+ + H) To a stirred, argon purged solution of the N-methyl benzyl urethane (1.00 g; 1.82 mmol) in 96:4 MeoH-Hco2H (25 mL) was added p~ lm black (0.37 g). The reaction mixture was stirrred for 25 16 h at ambient te.ll~e.~lul~. The catalyst was removed by filtration through Celite, and the filtrate solvents were removed under reduced pressure. The residue was purified by pressurized silica gel column chromatography using 95:5:0.5 CHC13:MeOH:NH40H as eluant. The N-methyl endo-amine product was obtained as a white foam.
3 TLC: Rf 0.35 (92:~:0.8 CHC13:MeOH:NH40H) HPLC (method A): retention time 7.~8 mir FAB MS: m/z 417 (M+ + H) To a stirred solution of the N-methyl endo-amine (0.650 g;
1.56 mmol) in CHC13 (~0 mL) was added the acid fluoride of Na-Boc-b'3~

L-methionine sulfolIe (510 mg; 1.80 mmol) and DIEA (0.35 mL; 2.0 mmol). The mixtul-e was stirred at ambient temperature for 48 h, and then extracted with 10% aqueous citric acid solution (2 x 30 mL), water (30 mL), and aqueous NaHCO3 (2 x 30 mL~. The organic phase was 5 dried (MgSO4), filtered, and the solvent was removed under reduced pressure. The residue was dissolved in CH2C12 (10 mL), and to the solution was added TFA (6 mL). The mixture was stirred at ambient U~ UI~ for 1.5 h. The solvents were removed under reduced pressure and the residue was purified by preparative reverse phase HPLC using a water-acetonitrile gradient containing 0.1% TFA. The TFA salt of the title compound was obtained as a Iyophili~ed powder.
Analysis calculated for (C2gH4sN3OsS~ 0.4 TFA-0.5 EtOAc C 57.0~ H 7.55 N 6.28 Found: C 57.0~ H 7.44 N 6.28 TLC: Rf 0.18 (95:5:0.5 CHC13:MeOH:NH40H) HPLC (method A): retention time 9.80 min FAB MS: m/~ 580 (M+ + H) IH NMR (300 MH;~, CDC13): d 7.15-7.25 (m, 4H), 5.20 (m, 2H), 3.17 20 (s, 3H), 2.92 (s, 3H), 1.06 (s, 3H), 0.96 (s, 3H) LX~MPLE 66 To a stirred solution of endo-(lS)-1'(((2-amino-7,7-25 dimethylbicyclo(2 2.1 )-hept- I -yl)-methyl)-sulfonyl)spiro( I H-indan-1,4'-piperidine (94 Ing; 0.17 mmol) in ethanol (20 mL) was added N-(3-bromopropyl)-phth~limi~1P (69 mg; 0.255 mmol) followed by diisopropylethyl amine (0.044 mL; 0.255 mm~ol). The temperature wa~
then increased to 50C. After i~ u~ lately 18 hr the ~olution was 30 cooled then concentrated under reduced pressure. The residue was dissolved in methyl~ne chloride (150 mL), washed with IM HCI (2 x 150 mL) then dried over sodium sulfate and concentrated. Purification by flash chromatography (5% methanol in methylene chloride) afforded the title compound (45 mg; 40%) as a white solid.

2~ ~46~û

Analysis ~ t~d for (C39H50N4O6S)-0-10 H20 0 15 CH2C12 C, 65.54; H, 7.10; N, 7.81 Found: C, 65.52; H, 7.09; N, 7.71 HPLC: (Vydac C18 Column; gradient from 95/5 to /100 H20/CH3CN
with 0.1% TFA. 15 min. gradient, flow rate = 1.5 ml/min.) Rt= 13.81 min. Purity= 100%
IHNMR: Consistent with structure FABMS: m/~ = 703 (M+ + H) 5 ~ >
S2 1~OH O O~,NH2 NH J~"N ~,~

Valine methyl ester (2.5g, 14.9 mmol), t-BOC-methionine (3.73 g,l4.9 mmol), HBT (2.01 g, 14.9 mmol) and EDC (2.33g, 14.91 mmol) were suspended in methylene chloride (60ml). The pH of the reaction was adjusted using E. Merck strips to about 9 with Et3N. After stirring ovemight at ambient ~ a~UI~, the reaction was evaporated and the residue was dissolved in ethyl acetate (60m~). The ethyl acetate solution was extracted three times with 5M citric acid (20 ml), three times with I M sodium bicarbonate (20 ml), one time with water and one time with saturated sodium chloride (20ml). The ethyl acetate solution was then dried over anhydrous sodium sulfate, filtered and evaporated to dryness.
The residue was dissolved in methyl iodide (25ml) and stirred overnight, and the resulting mixture was evaporated to dryness.
The resultin~ methyl sulfonium salt (4.72mmol) was dissolved in a 1:1 .... . . .. . . ... . . . . .. , ... ... , . .. , . . . .. . _ WO 9~i114025 PCrlUS94/13483 2~ 5~

mixture of DMF and methylene chloride (150ml). After cooling to zero degrees in a saltwater/ice bath, sodium hydride (60% in mineral oil:
0.57g, 9.44mmol) ~ivas added. After stirring for 3hr, the reaction was treated with glacial acetic acid (I ml) followed by water (I ml). The 5 mixture was stirred for 15min. at ambient temperature, then evaporated. The crllde product was dissolved in water (150 ml) and neutralized with aqlleous sodium bicarbonate. The mineral oil and neutral by-products were removed by extracting three times with methylene chloride (50 ml). The aqueous layer was then acidified with citric acid and extracted three times with methylene chloride (50ml).
The combined metJtlylene chloride layers were dried over sodium sulfate, filtered and evaporated. Purification of the crude product was carried out by prep~lrative HPLC using a Waters C-18 column in a SepTech ST/Lab 800C ill~llUIll~lll with a water/acetonitrile gradient 15 increasing from lO'Yo to ca. 60% acetonitrile.
The pr~duct so obtained (60 mg, 0.20 mmol) was combined with (lS)~ (((7,7-dimethyl-(2-endo-aminomethyl-2-exo-hydroxy)-bicyclo-(2.2. 1 )-hept- I -yl)-methyl)sulfonyl)-spiro-( I H-indene- 1,4'-piperidine) (86 mg, 0.20 rnmol), HBT (27 mg, 0.20 mmol) and EDC
20 (38 mg, 0.20 mmol) in methylene chloride (10 ml). The pH of the mixture was adjust~d to ca. 9 with triethylamine. After stirring overnight, the reaction was evaporated and the residue was dissolved in ethyl acetate (25ml). The ethyl acetate solution was extracted three times with 5M citric acid (2ml), three times with IM sodium 25 bicarbonate (2ml), ~ne time with water and one time with saturated sodium chloride (2ml). The ethyl acetate solution was then dried over anhydrous sodium sulfate, filtered and evaporated to dryness. The product was dissol~ed in a 40% solution of trifluoroacetic acid in methylene chloride (3 ml) and stirred for Ihr at ambient temperature.
30 The solvent was then evaporated, and the residue was dissolved in methanol and puri~led by preparative HPLC using the conditions described.
NMR: Consistent with structure FAB MS: 613 (M+ + H) HPLC: >99% pure at 214 nM
Analysis calc'd for C33H47N405 2.0 C2HF3o2 l.4H2o:
Calc'd: C, 51.31; H, 6.15; N, 6.47 Fourld: C, 51.31; H, 6.00; N, 6.66.

10 ~ "
SO2 ~~,~

To a solution of (IS)-1'-(((7,7-dimethyl-(2-endo-amino-methyl-2-exo-hydroxy)-bicyclo-(2.2. 1 )-hept- I -yl)-methyl)sulfonyl)-spiro(lH-indene-1,4"-piperdine (100 mg; 0.23 mmol) in DMF was added t-boc-L-valine (60 mg; 0.28 mmol), I-hydroxybenzotriazole (37 mg; 0.28 mmol), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide 25 (53 mg; 0.28 mmol). The pH was adjusted to 9 with triethylamine (96 uL; 0.69 mmol). After stirring for 1.5 hrs., the solution was con~erltr~l~d to dryness and treated with 5% citric acid solution. This water layer was then basified with saturated sodium bicarbonate and extracted 3 times with ethyl acetate. The organics were combined and 3C dried over sodium sulfate, filtered, and concentrated to yield a white oily foam. This white foam wa~ dissolved in ethyl acetate, cooled to 0C and treated with saturated sollution of HCI in ethyl acetate. The reaction mixture was stirred for 45 min., purged with N2 for 30 min., and then concentrated to dryness. The resulting solid was partitioned between ethyl acetate and ~aturated ~odium bicarbonate. The a~lueous 2 1 l4~0 layer was extracted 3 times with ethyl acetate, and the combined organic fractions were dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was puri~led by flash chromatography (eluted witll 98/2/0.2 of methylene chloride/methanol/ammonium hydroxide) to yield 5 the title compound as a white foam.
m.p.: not available NMR: consistent ~vith structure HPLC > 97% pure 0 MS MH-530.3 ~AB)]
CHN Analysis calculated for C29H43N3O4S
C, 65.75; H, 8.18; N, 7.93 Found: C, 65.75; H, 8.18; N, 7.93 2 ¦~OH o ~NH~

To a solution of (IS)-1'-(((7,7-dimethyl-(2-endo-amino-methyl-2-exo-hydr~;)xy)-bicyclo-(2.2.1)-hept-1-yl)-methyl)sulfonyl)-spiro(lH-indene-l,~"-piperdine) 51 mg; 0.12 mmol) in DMF was added 30 t-boc-L-leucine (36 mg; 0.14 mmol), I-hydroxybenzotraizole (19 mg;
0.14 mmol), and 1 -(3-dimethylaminopropyl)-3-ethylcarbodiimide (28 mg; 0.14 mmol). The pH was adjusted to 9 with triethylamine (50 ~LL, 0.36 mmol). After stirling for 18 hrs., the solution was concentrated to dryness and treated with 5% citric acid solution. The water layer was then basified with .saturated sodium bicarbonate and extracted 3 times WO 9!5/14025 PCr/US94/13483 21 74651~

with ethyl acetate. The organics were combined amd dried over sodium sulfate, filtered, and concentrated to yield a yellow oil. This yellow oil was dissolved in ethyl acetate, cooled to 0C and treated with a saturated solution of HCl in ethyl acetate. The reaction mixture was stirred for 5 45 min., purged with N2 for 30 min., and then concentrated to dryness.
The resulting solid was partitioned between ethyl acetate and saturated sodium l,:cd~ atc the aqueous layer was extracted 3 times with ethyl acetate and the combined organic fractions were dried over sodium sulfate, filtered, and concentrated in vacuo. The residue purified by flash chromatography (eluted with 98/2/0.2 of methylene chioride/methanol/qmmnnillm hydroxide) to yield the title compound as a white foam.
m.p.: 88 - 110 C
15 NMR: consistent with structure HPLC: > 98% pure MS: MH-544.3 (FAB) CHN Analysis cq1~ 1l1qt~d for C30H4sN304SØ35H20:
C, 6550; H, 8.37; N, 7.64 Found: C, 65.48; H, 8.39; N, 7.60 SO2 ~OH O O~ NH2 3 0 ~ NJ~
To a ~olution of (lS)-1'-(((7,7-dimethyl-(2-endo-amino-methyl-2-exo-hydroxy)-bicyclo-(2.2. 1 )-hept- I -yl)-methyl)sulfonyl)-spiro(lH-indene-1,4"-piperdine) (72 mg; 0.17 mrnol) in DMF was 21 74~50 added 2-(3-amino-2-oxopyrrolidin-1-yl)acetic acid (52 mg; 0.20 mmol), 1-hydroxybenzotria~ole (27 mg; 0.20 mmol), and 1-(3-dimethyl-aminopropyl)-3-ethylcarbodiimide (38 mg; 0.20 mmol). The pH was adjusted to 9 with triethylamine (70 IlL; 0.50 mmol). After stirrin~ for 5 18 hrs., the solution was concentrated to dryness and treated with 5%
citric acid solution. The water layer was then basified with saturated sodium bicarbonate and extracted 3 times with ethyl acetate. The organics were combined and dried over sodium sulfate, filtered, and concentrated to yield a yellow oil which was purified by flash o chromatography (eluted with 98/2/0.2 of methylene chloride/met~lanol/
ammonium hydroxide). The product was dissolved in ethyl acetate, cooled to 0C, and treated with a saturated solution of HCI in ethyl acetate. The reaction mixture was concentrated to dryness. The resulting solid was partitioned between ethyl acetate and saturated 5 sodium bicarbonate. The aqueous layer was extracted 3 times with ethyl acetate and the organic fractions were combined, and dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by flash chromato~raphy (eluted with 9713/0.3 of methylene chloride/methanol/ammonium hydroxide) to yield the title compound a~
20 a white foam.
m.p.: 115- 155C
NMR: consistent with structure HPLC: > 98% pur~
25 MS: M+H-571 (FAB) CHN Analysis c~lr~ d for C3oH42N4o5s.o.8sH2o-o~25 C4HIoO:
C, 61.58; H, 7.70; N, 9.27 Found: C, 61.56, H, 7.44; N, 9.23 E~A~PLE 7 1 -WO9~/14025 2 1 74 ~5~ PCINS94113483 ~\ N ~
0 H N'~<
,~0+
o Endo ( 1 S)- I '(((2-~minomf~thyl-7 ,7-dimethylbicyclo-(2.2.1 )hept- I -yl)methyl)sulfonyl)spiro(indene- 1 ,4'-piperidine) (337 mg, 5 0.78 mmole) and N-isobutyl-oxycarbonyl-5,5-dimethyl-L-thiazolidine-4-carboxylic acid (230 mg, 0.90 mmole) were combined with 438 mg (0.99 mmole) of benzotriazol-l-yloxytris(dimethylamino)phosphonium hexafluorosphosphate in 5 ml of dry dimethyl formamide at room ,e-~lul~ with protection from moisture (calcium sulfate drying 20 tube). The pH of mixture was adjusted to 8.5 with diisopropyl-ethylamine. The resulting solution was allowed to stir at room e-dlul~ for two hours. The dimethyl~llllalllide wa3 then removed under reduced pressure, and the residue taken up in 150 ml of ethyl acetate and washed with saturated sodium bicarbonate (2 x 25 ml) and 25 brine (I x 25 ml). The ethyl acetate was dried over sodium sulfate, filtered, and ~O~ la~ed to give 941 mg of an oil. The analyticâlly pure material was obtained via chromatography on precoated silica gel plates (1.5 mm x 20 x 20 cm) developed with ethyl acetate in hexane (35:65). The product was obtained as an oil.
NMR: consistent with structure FAB MS: M++H=574 (Compound minus t-BOC) Analysis calculated for C35H51N3O6S2-1.3 CHC13 C, 52.58; H, 5.35; N, 5.07 Found: C, 52.62; H, 6 ,40; N, 4.99 WO 9~/14025 PCI/IIS94/13483 21 74~G

EX~MPLE 72 SO~-->H
N J~S~
H NH
The product of the preceding Example 71 was dissolved in ethyl acetate, and the solution was cooled to 0C. HCI (g) was bubbled into the solution for three minutes, and then let stand at 0C for 30 mirlutes. The ethyl acetate was removed under vacuum and the residue 20 evaporated twice from ethyl acetate. The residue was chromatographed on precoated silica gel plates 6.5 mm x 20 cm x 20 cm) using chloroform-methanol-ammonium hydroxide (97-3-0.3/v-v-v) to develop the plates, The product was obtained as an amophous solid.
25 m p 126 -129C
NMR: consistent with structure FAB MS: M++H = 574.3 Analysis calculated for C30H43N304S2-0.15 CHC13 C,61.19;H,7.35;N,7.10 30 Found: C, 61.]0; H, 7.33; N, 7.12 WO 95/14025 PC~rlUS94113483 21 74~5a [~ ~OH
S2 H J~ N~
H
To a solution of (lS)-1'-(((7,7-dimethyl-(2-endo-amino-methyl-2-exo-hydroxy)-bicyclo-(2.2. 1 )-hept- I -yl)-methyl)sulfonyl)-spiro(lH-indene-1,4-piperdine) (108 mg; 0.25 mmol) in degassed DMF
was added 4-imidazoleacrylic acid (41.44 mg; 0.30 mmol) and benzotriazol-l-yloxy-tris(dimethylamino)phosphonium hexafluorophosphate (132.69 mg; 0.30 mmol). The pH of the reaction mixture was adjusted to 9 with diisopropylethylamine. After stirring for 14 hr, the reaction mixture was concentrated to dryness. The resulting solid was dissolved in methanol and then purified by reverse phase preparative HPLC utilizing a Water C-18 column (45 minute gradient, 5% to 70% acetonitrile/water containing 0.1% trifluoroacetic acid, 13.5 ml/min flow rate). The product containing fractions were combined and Iyophilized to give an amorphous white solid.
NMR: consistent with structure;
HPLC: > 97% pure (254 nM) FAB MS: M+H+ thioglycerol matrix = 659 CHN Analysis calculated for C30H38N4o4s-l~6cF3co2H
Calc'd: C, 54.39; H, 5.44; N, 7.64.
Found: C, 54.50; H, 5.20; N, 7.53.
In addition to those compounds specifically exemplified above, additional compounds of the present invention are set forth in tabular forrn belo~. These compounds are synthesized by use of the synthetic routes and methods described in the above Schemes and 21 7~50 Exarnples and variations thereof well known to those of ordinary skill in the art, and not requiring undue e~ ion. All variables listed in the Tables belo~v are with reference to the following generic structure:
T~BLES
10 ~
R= (CH2) I R~R6 ~CH2)p =~ 2)q L\
Variable 0 95/1402~; PCT/US94/13483 ~74G~

TABLE I
Variable=
NH,SO2~ R H~J~F

NH 'S2\
R` SO2 H 1~' NH R~N~
o NH ~
R~NH~SO2\¢~ R N~3 21 7~650 TA.BLE I (~ONT'D) 5 R, ,so2~ ,J~I H
R NH~~ R~
10 o OH
--NH J~ NH~OCH3 --NH~3 R N~

NH~--~ R~
N~ ,~Br NH~

R ~ N ~f ~CI N HJ~N

Cl'l~S94/13483 WO 95/1402!i P
21 7~65~

O O ~
\--NHJ~ NHJ~ J~J
\--NHJ~ H~

`N~ R~N~CI
R`NJ~OCH3 NJ~ J~OCH3 H
--NH~ R ~OCH3 21 74~5~ ~

TABLE 2 CONT'D
R NH2 R ,~
0 R~--CI N~1113 --N~--\/ R~CO2H
H~f~No2 CH3 R ~ N ~3 R N

WO gS/14025 PCI~/US94/1348 21 7~6~

TABI,E 3 R ~ N ~S2~/~ R --~
H ~`OCH3 H
OH --H~
10 ~ O
N~J~OH ~N
O
o R`f NH2 NH~COOH N~

r WO 95/14025 PCI/US941134~3 2~7~

TABLE 3 CONTrD
R~,N~--COOH ~ N~NH
H CH3 ~ ~ R~N~
NH
R~,COOH ~'H~
R~ O`/ `J'` NH2 H~

2l 7~55 5 NH~ --HJ~
¢~0 0 O~,NH2 R J~ N~ ~H
~ ~ NH
R N ~ C H3 -2~ 746~0 RLF 4 CONT'~
~,~ R N~ ,NH2 O R ~ N ~,CO2H
R N)~ o HCO2~ R N~
O ~ O SO3H
RJ~Hi N
N ~ HN J~----N

WO 9~/14025 2 1 7 4 6 3 0 PCI'IUS94/13483 TABLE S
R~N J~o~0 R~,N~
R~N ~,~ S J~s><
15 H oHcHis ~ o R~N~
O COOH

R~ N ~?N R~ N~ +~

3 o H~ 3 R~N~N~CO2H

WO 9~/14025 PCT/US94/13483 21 /4~0 o N ~ ~ \~ X NH

OH

NH ~ R~ ~=
O COOC,~, R~ Q~=¦

O o ~
~ NH J~N~ NH2 R~ N J~
OH
H J~ J~l OH `~S

R~ J~N~
sl ~o J~ N J~S

15 R~NH~--Nf'l~S~o R`N'~ NH2 o=( N H2 0~

O O
O COOCH3 NH NHJ~O'¦\
NH\I~o~<
R~ ~X~H3O ` NH/J~ N H

R~N ~NH2 2 ~ 3 ~

5 ~ J3 R`N~NH2 NH~J o~S~
10 o O R`
R~ J~NHJ~O~ NHJ~ X
S~ o, CO2CH3 NH/~HJ`~

R~N /~\N~ NH N

21 7~5G

R~ NH~ R~ '~N~?
NH2 o o R~ J~\'^`OH R~ ~N~
NH I R~NH~N
R "S"O R~ NH--~O~
-S~ ~`S~;
R~NH~ J~ ~NH2 H
~f ~
NH

WO 95/14025 PCI'NS94/13483 21 74~50 TABI~E~ 10 R J~, N ~X
R "S"O R~N~
R~ ~N N~/~N~
o,S ~o o NH~--NH ~
R~H~X ~~
SO~2 R~ ~NH2 R\~N~ NH ~--~ NH ~S
O O

WO 9~/1402~ PCr/US94/13483 21 74~50 RADIOLICAND BINDING ASSAYS
The high affinity binding of [3H]oxytocin (OT) to uterine 5 tissue and [3H]arginine vasopressin (AVP) to liver (AVP-Vla site) and kidney (AVP-V2 site) tissue was determined using crude membrane preparations as described previously [Pettibone, D.J., et al., J.
Pharmacol. and Exper. The7-., 256(1): 304-308 (1991)]. Uterine tissue was taken from nonpregnant adult Sprague-Dawley rats (Taconic Farms, Germantown, NY) pretreated (18-24 h) with diethylstilbestrol propionate (DES; 300 llg/kg, i.p.). Uterine tissue (full thickness) was also taken with informed consent from nonlabor pregnant women undergoing cesarean section at 38 to 39 weeks gestation (Oregon Health Sciences Center, Portland, OR). Liver and kidney medulla samples 5 were taken from male rats and from human surgical and early postmortem donors (National Disease Research Interchange, Philadelphia PA; Analytical Biological Services, Wilmin~tr)n, DE).
Competition studies were conducted at equilibrium using 1 nM [3H]oT or 0.5 nM [3H]AVP in the following buffer: 50 mM Tris, 20 5 mM MgC12, 0.1% bovine serum albumin. Nonspecific binding was ~ t~rminf d using I ~lM unlabeled OT or AVP in their respective assay~.
The binding reactions were initiated by the addition of tissue preparation and terminated by filtration using a Skatron cell harvester (model 7019, Skatron, Inc., Sterling, VA). Ki values were calculated 25 for each compound using three to six separate IC50 d~ dlions (K;=lCso/[l- c/Kd]); [Cheng, Y-C; Prusoff, W.H.; Biochem Pharmacol 22:3099 (1973)] with mean Kd values obtained from replicate (n = 3) equilibrium saturation binding assays (10 point, 100 fold concentration range): [3H]oT rat utenus, 0.69 nM; human myometrium, I.l nM;
30 [3H]AVP: rat liver, 0.21 nM; rat kidney, 0.27 nM; human liver, 0.27 nM; human kidney, 1.4 nM. Computer analysis of the saturation assays by EBDA/LIGAND [McPher.son, G.A.; Kinetic, Ebda, Ligand, Lowry:
A Collection of Radioligand Binding Analysis Programs, Elsevier Science Publishers, Amsterdam ( 1985)] indicated that both radioligands apparently bound to single sites in all tissues examined. The final WO 95/14025 PCr/US94113483 2 i 7 ~ b ~ G

protein concentration for the various tissues in each assay ranged from 150 to 300 ,ug/ml [Lowry, P.H.; Rosebrough, N.J.; Farr, A.L.; Randall, R.J.; J. Biol. Chem., 193 265-275 (1951)].
IC50 ~alues were determined for the [3H]oT and [3H]AVP
5 binding assays by linear regression of the relation log concentration of compound vs. percent inhibition of specific binding. Data is either reported as a given percentage of inhibition at a specified concentration, or if an IC50 was c~ r~-l, as a nanomolar concentration.
Rc;~ e IC50 values of the compounds of the instant invention for rat L3H]oT are given below.
l~xample Result For l_HlOT
70% inhib. at 1000nM
2 1100nM
3 50% inhib. at 1000nM
4 620nM
5 190nM
6 60% inhib. at 1000nM
20 7 54% inhib. at 1000nM
8 140nM

9 68% inhib. at 1000nM

10 140nM
I 1 69% inhib. at 1000nM
2512 37% inhib. at 100nM
13 1 80nM
14 76% inhib. at 1000nM
15 160nM
16 61nM
3017 45nM
1~ 52% inhib. at 100nM
19 150nM
20 50% inhib. at 100nM
21 61nM

-- 21 74~5~

Example Result For l-HlOT
22 58.8nM
26% inhib. at 10000nM
26 34% inhib. at 1000nM
27 47nM
28 82nM
29 31nM
45nM
32 36% iohib. at 10000nM
33 30% inhib. at 1000nM
34 48% inhib. at 1000nM
43% inhib. at 1000nM
36 67nM
38 33% inhib. at 1000nM
39 8500nM
71% inhib. at 10000nM
41 120nM
42 96.5nM
43 4300nM
44 39% inhib. at 100nM
25% inhib. at 100nM
46 36% inhib. at 100nM
47 4600nM
48 90nM
49 54nM
so 71% irlhib. at 1000nM
51 84% inhib. at 1000nM
52 290nM
67 7.6 nM
73 45 nM

2l 74~5~ --Binding (Ki) of compoumds to Hulnan OT, Human AVP-VIa and Human AVP-V2:
~xample OT AVP-V I a AVP-V~
s 67 ~~ 3.7 nM 1.4 nM 1600 nM
73 3.8nM 2.7 nM --While the invention has been described and illustrated wit~
reference to certail1 preferred embodiments thereof, those skilled in th~
art will appreciate that various changes, modifications and substitutions can be made therein without departing from the spirit and scope of the invention. For example, effective dosages other than the preferred dosages as set forth hereinabove may be applicable as a consequence of variations in the responsiveness of the mammal being treated for prevention of preterm labor, or for other indications for the compounds of the invention indicated above. Likewise, the specific pharmacological r~sponses observed may vary according to and depending upon the particular active compound selected or whether there are present pharmaceutical carriers, as well as the type of 20 formulation and mode of administration employed, and such expected variations or differences in the results are contemplated in accordance with the objects and practices of the present invention. It is intended, therefore, that the invention be limited only by the scope of the claim~
which follow and that such claims be interpreted as broadly as is 25 reaSO!lable-

Claims

Claims #1-23 have been cancelled.

24. A method of treating preterm labor in a mammal in need thereof, comprising administering to said mammal a pharmacologically effective amount of a compound which binds to a human oxytocin receptor with a binding affinity which is no more than ten-fold higher or ten-fold lower than the binding affinity with which the compound binds to a human arginine-vasopressin-V1 (AVP-V1a) receptor.
25. The method of Claim 24, wherein the compound binds to the human oxytocin receptor with a binding affmity which is no more than five-fold higher or five-fold lower than the binding affinity with which the compound binds to the human AVP-V1a receptor.
26. The method of Claim 25, wherein the compound is selected from or 27. A method of treating dysmenorrhea in a mammal in need thereof, comprising administering to said mammal a pharmacologically effective amount of a compound which binds to a human oxytocin receptor with a binding affinity which is no more than ten-fold higher or ten-fold lower than the binding affinity with which the compound binds to a human arginine-vasopressin-V1 (Avp-V1a).
receptor.
28. The method of Claim 27, wherein the compound binds to the human oxytocin receptor with a binding affinity which is no more than five-fold higher or five-fold lower than the binding affinity with which the compound binds to the human AVP-V1a receptor.

29. The method of Claim 28, wherein the compound is selected from or .

30. A compound selected from the group consisting of and wherein R is selected from , , , , , , .

, , , , or ;
and R' is selected from , , , , or and the pharmaceutically acceptable salts thereof.

31. A compound of the formula wherein R is selected from , , , , , or and the pharmaceutically acceptable salts thereof.

32. The compound of Claim 31 selected from or and the phamaceutically acceptable salts thereof.

33. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a pharmacologically effective amount of a compound as claimed in Claim 32.
34. A method of preventing preterm labor in a mammal in need thereof, comprising the step of administering to said mammal a pharmacologically effective amount of a compound as claimed in Claim 32.
35. A method of treating dysmenorrhea in a mammal in need thereof, comprising the step of administering to said mammal a pharmacologically effective amount of a compound as claimed in Claim 32.