Note: Descriptions are shown in the official language in which they were submitted.
WO 94/1 1012 PCI~/US93/105~1
~1 47503
TITLE OF THE INVENTION
SUBSTITUTED DIPEPTIDE ANAL~GS PROMOTE RELEASE OF
GROWTH ~IORMONE
5 BACKGROUND OF THE INVENTION
Growth hormone, which is secreted from the pituitary,
stim~ tes growth of all tissues of the body that are capable of growing.
In addition, growth ho~none is known to have the following basic
effects on the metabolic process of the body:
1. Increased rate of protein synthesis in all cells of the body;
2. Decreased rate of carbohydrate lltili7~tion in cells of the
body;
3. Increased mobilization of free fatty acids and use of fatty
acids for energy.
A deficiency in growth hormone secretion can result in
various medical disorders, such as dwarfism.
Various ways are lcnown to release growth hormone. For
example, chemicals such as arginine, L-3,4-dihydroxyphenylal~nine
(L-DOPA), glucagon, vasopressin, and insulin induced hypoglycemia, as
20 weli as activities such as sleep and exercise, indirectly cause growth
hormone to be released from the pituitary by acting in some fashion on
the hypoth~l~mll~ perhaps either to decrease somatostatin secretion or to
increase the secretion of the known secretagogue growth hormone
releasing factor (GRF) or an unknown endogenous growth hormone-
25 releasing hormone or all of these.
In cases where increased levels of growth hormone weredesired, the problem was generally solved by providing exogenous
growth hormone or by ~rlmini~tering an agent which stiml~l~ted growth
hormone production and/or release. In either case the peptidyl nature
30 of the compound necessitated that it be ~1mini~tered by injection.
Initially the source of growth hormone was the extraction of the
pituitary glands of cadavers. This resulted in a very expensive product
and carried with it the risk that a disease associated with the source of
the pituitary gland could be transmitted to the recipient of the growth
WO 94/11012 PCI/US93/}05~1
? 5 L~
hormone. Recently, recombinant growth hormone has become available
which, while no longer carrying any risk of disease tr~n.cmi.~sion, is still
a very expensive product which must be given by injection or by a nasal
spray.
Other compounds have been developed which stimulate the
release of endogenous growth hormone such as analogous peptidyl
compounds related to GRF or the peptides of U.S. Patent 4,411,890.
These peptides, while considerably smaller than growth hormones are
still susceptible to various proteases. As with most peptides, their
potential for oral bioavailability is low. The ins~ant compounds are
highly substituted dipeptide analogs for promoting the release of growth
hormone which are stable under various physiological conditions which
may be ~clmini~tered parenterally, nasally or by the oral route.
SUMMARY OF THE INVEN~ION
The instant invention covers certain substicuted dipeptide
analogs which have the ability to stim~ te the release of natural or
endogenou~ gr~wth h~mlone. The compolmds thlls have ~e ability to
be used to treat conditions which require the stiiinulation of growth
20 hormone production or secretion such as in hllm~n~ with a deficiency of
natural growth hormone or in ~nim~l~ used for food production where
the stim~ tion of growth hormone will result in a larger, more produc-
tive ~nim~l. Thus, it is an object of the instant invention to describe the
diphenyl substituted dipeptide analogs. It is a further object of this
25 invention to describe procedures for the preparation of such com-
pounds. A still further object is to describe the use of such compounds
to increase the secretion of growth hormone in hurnans and ~nim~l~. A
still further object of this invention is to describe compositions
cont~ining the substituted dipeptide analogs ~or the use of treating
30 hllm~n~ and ~nim~l~ so as to increase the level of growth hormone
secretions. Further objects will become apparent from a reading of the
following description.
WO 9~/11012 PCr/US93/10551
-- - 214~3
DESCRIPTION OF THE INVENTION
The novel substituted dipeptide analogs of the instant
invention are best described in the following structural formula I:
(X)n
la
R2a~rl=~ O(C I 2)p ,R4
l O R3a~ (L)w--(C H2)q--N--C--C H--N--C--A--N~ R5
where L is
,~/
3b
~/ ~J R
1b~ R2b
20 nisOorl;
p is O to 6;
q is O to 4;
w is O or 1 ;
OH R1 O
X is C=O, O, S(O)m, -CH-, -N-, -CH=CH-;
m is O to 2;
30 Rl, R2, R1a, R2a, Rlb, and R2b are independently hydrogen, halogen,
Cl-C7 alkyl, C1-C3 perfluoroalkyl, Cl-C3 perfluoroalkoxy, -S(O)m
R7a, cyano, nitro, R7bo(cH2)v-~ R7bCoo(cH2)v-7 R7boco-(cH2)
R4R5N(CH2)V-, R7bCoN(R4)(cH2)v-~ R4R5NCo(cH2)v-~ R4R5-
NCOO(CH2)V-, phenyl or substituted phenyl where the substituents are
from 1 to 3 of halogen, Cl-C6 alkyl, Cl-C6 alkoxy, or hydroxy; R7a
WO 9~1/11012 PCI/US93/~05~1
and R7b are independently hydrogen, Cl-c3 perfluoroalkyl, Cl-C6
aLkyl, substituted Cl-C6 alkyl, where the substitu.ents are phenyl or
substituted phenyl; phenyl or substituted phenyl where the phenyl
substituents are from 1 to 3 of halogen, C1-C6 alkyl, C1-C6 alkoxy, or
5 hydroxy and v is O to 3;
R3a and R3b are independently hydrogen, R9, Cl-C6 alkyl substituted
with R9, phenyl substituted with R9, or phenoxy substituted with R9;
o R9 is
N--N N--N
,N ~ " ~a
R4a
R7bo(CH2)V-, R7bCoo(cH2)v-~ R7boCo(cH2)v-~ R7bCo(CH2)V-,
R7bo(cH2)vco-~ R4bR 1 2CN(CH2)V -, R 1 2aR 1 2~NCO(CH2)v-,
R12aR12bNCS(CH2)v, R4bRl2aNN(Rl2b)co(cH2)v-
~R4bR 1 2aNN(R 1 2b)cs(cH2)v-~ R4bR 1 2aNcoN(R 1 2C)(CH2)V-,
2 R4bR 1 2aNCSN(R 1 2C)(CH2)V-, R4bR 1 2aNN(R 1 7~b)CoN(R 1 2C)(CH2)V-
,R4bR 1 2aNN(R 1 2b)CSN(R 1 2c)(cH2)v-~ R4bR 1 ~aNN(R 1 2b)
COO(CH2)V-, R4bR12aNCoo(cH2)v-~ or R130CoN(Rl2c)(cH2)
where v is O to 3;
25 ~12a, R12b and R12c are independently RSa~ ORsa~ or CoRsa; R12a
and R12b, or R12b and R12C, or Rl2a and Rl2c~ or R4b and R12a, or
R4b and Rl 2a~ or R4b and Rl2c~ or Rl 3 and Rl 2c~ can be taken
together to form -(CH2)r-B-(CH2)S- where B is cHRl~ 0, S(O)m or
NRlO~ m is 0, 1 or 2, r and s are independently O to 3 and R1 and Rl 0
30 are as defined;
R13 is Cl-C3 perfluoroalkyl, Cl-C6 alkyl, substituted Cl-C6 alkyl,
where the substituents are hydroxy, -NR lOR 1 1, carboxy, phenyl or
substituted phenyl; phenyl or substituted phenyl ~vhere the substituents
WO 94/11012 PCr/US93/10551
2~503
on the phenyl are from 1 to 3 of halogen, Cl-C6 alkyl, Cl-C6 alkoxy or
hydroxy where RlO and Rl l are as defined;
R14 is hydrogen, Rl, R2 independently disubstituted phenyl, Cl -ClO
5 alkyl or substituted Cl-Clo alkyl where the substituents are from 1 to 3
of imidazolyl, indolyl, hydroxy, fluoro, S(o)mR7a~ Cl-C6 alkoxy, C3-
C7 cycloalkyl, R1, R2 independently disubstituted phenyl, Cl-C3
alkoxy, R 1, R2 independently disubstituted phenyl Cl -C5 alkanoyloxy,
Cl-C5 alkoxycarbonyl, carboxy, formyl or -NRloRl 1 where Rl, R2,
10 and R1 l are as defined;
Rl5 is hydrogen, trifluoromethyl, Rl, R2 independently disubstituted
phenyl, Rl, R2 independently disubstituted naphthyl, C3-C7 cycloalkyl,
Cl-C1o alkyl, substituted Cl-C1o alkyl where the substituents are from
5 1 to 3 of hydroxy, fluoro, S(o)mR7a~ C1-C6 alkoxy, C3-C7 cycloalkyl,
R1, R2 independently disubstituted phenyl, R1, R2 independently disub-
stituted phenyl C1-C3 alkoxy, R1, R2 independently disubstituted
naphthyl, Rl, R2 independently disubstituted naphthyl Cl-C3 aLkoxy,
Cl-C5 alkanoyloxy, Cl-C5 alkoxycarbonyl, carboxy, formyl,
2 -NR l OR l l or R l, R2 independently disubstituted heterocycle, where the
heterocycle is imidazole, thiophene, furan, pyrrole, oxazole, thiazole,
triazole, tetrazole, pyridine, benzofuran, benzothiophene, benzimi-
dazole, indole, 7-azaindole, oxindole or indazole; where Rl, R2, RlO
and Rl l are as defined above;
R4, R4a, R4b, R5 and R5a are independently hydrogen, phenyl,
substituted phenyl, C1 -Clo alkyl, substituted C1 -ClO alkyl, C3-clo
alkenyl, substituted C3-Clo alkenyl, C3-C1o alkynyl, or substituted C3-
Clo aL~cynyl where the substituents on the phenyl, alkyl, alkenyl or
30 alkynyl are from l to 5 of hydroxy, C1-C6 alkoxy, C3-C7 cycloalkyl,
fluoro, R1, R2 independently disubstituted phenyl Cl-C3 aIkoxy, R1,
R2 independently disubstituted phenyl, Cl-C20-alkanoyloxy, C l-C5
alkoxycarbonyl, carboxy, fo~nyl, or -NR l OR 1 1 where R l O and R 1 1
are independently hydrogen, Cl-C6 alkyl, phenyl, phenyl Cl-C6 alkyl,
WO 9~/11012 PCI/US93/105~1
- 6 -
- C1-Cs-alkoxycarbonyl, C1-Cs-alkanoyl or C1-C6 alkyl; or R4 and R5
can be taken together to form -(CH2)rB(CH2)S- where B, r, s, Rl and
R10 are as defined above;
R6 is hydrogen, Cl-C1o alkyl, phenyl or phenyl C1-C1o alkyl;
A is
~8
o --(CH2)x--C--(CH2)y
R8a
where x and y are independently 0-3;
R8 and R8a are independently hydrogen, Cl-Clo alkyl, trifluoromethyl,
phenyl, substituted Cl-Clo aLkyl where the subs~ituents are from 1 to 3
of imidazolyl, indolyl, hydroxy, fluoro, S(o)mR7a~ Cl-C6 aLkoxy, C3-
C7 cycloalkyl, Rl, R2 independently disubstitute~ phenyl Cl-C3 alkoxy,
Rl, R2 independently disubstituted phenyl, Cl-Cs-alkanoyloxy, C1-Cs
20 alkoxycarbonyl, carboxy, formyl, or ~ oRl 1 where R10 and Rl 1
are as defined above; or R8 and R8a can be taken together to form
-(CH2)t- where t is 2 to 6; and R8 and R8a can independently be joined
to one or both of R4 and RS to form alkylene bridges between the
terminal nitrogen and the alkyl portion of the A group wherein the
25 bridge contains from 1 to 5 carbon atoms;
and pharmaceutically acceptable salts thereof.
In the above structural formula and throughout the instant
specification, the following terms have the indicated meanings:
The alkyl groups specified above are intended to include
3 o those alkyl groups of the designated length in either a straight or
branched configuration. Exemplary of such alkyl groups are methyl,
ethyl, propyl, isopropyl, butyl, sec-butyl, tertiary butyl, pentyl,
isopentyl, hexyl, isohexyl, and the like.
The alkoxy groups specified above are intended to include
those alkoxy groups of the designated length in either a straight or
WO 94/11012 PCr/US93/10551
~ 2147~03
branched configuration. Exemplary of such alkoxy groups are
methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, tertiary
butoxy, pentoxy, isopentoxy, hexoxy, isohexoxy and the like.
The term "halogen" is intended to include the halogen atom
5 fluorine, chlorine, bromine and iodine.
Certain of the above defined terms may occur more than
once in the above formula and upon such occurrence each term shall be
defined independently of the other.
Preferred compounds of the instant invention are realized
o when in the above structural formula:
nisOor1;
p is O to 4;
q is O to 2;
15 wisOorl;
R10
I
X is O, S(O)m, -N-, -CH=CH-;
20 misO~o2;
Rl, R2, Rla~ R2a7 Rlb~ and R2b are independently hydrogen, halogen,
C1-C7 alkyl, Cl-C3 perfluoroalkyl, -s(o)mR7a~ R7bo(CH2)v-,
R7bCoo(cH2)v-~ R7boco(cH2)v-~ phenyl or substituted phenyl where
25 the substituents are from 1 to 3 of halogen, Cl-C6 alkyl, Cl-C6 alkoxy,
or hydroxy;
R7a and R7b are independently hydrogen, Cl-c3 perfluoroalkyl, C1-C6
alkyl, substituted C1-C6 alkyl, where the substituents are phenyl; phenyl
30 andvisoto2;
R3a and R3b are independently hydrogen, R9, Cl-C6 alkyl substituted
with R9, phenyl substituted with R9, or phenoxy substituted with R9;
WO 9 1/1 1012 PCI/US93/105~1
,3
- 8 -
R9 is as defined above;
R12a, Rl2b and Rl2c are independently RSa~ OR Sa~ or CoR5a; R12a
and R12b, or R12b and R12C, or R13 and R12b or R12a and R4b can be
5 taken together to form -(cH2)r-B-(cH2)s- where B is CHR1, O, S(O)m
or NR10~ m is 0, 1 or 2, r and s are independently 0 to 3, R1 is as
defined above and R10 is hydrogen, C1-C6 alkyl, phenyl C1-C6 alkyl or
C1 -C5 alkanoyl-C1 -C6 alkyl;
o R13 is C1-C3 perfluoroalkyl, Cl-C6 alkyl, substituted C1-C6 alkyl,
where the substituents are hydroxy, NRloRl 1, carboxy, phenyl or
substituted phenyl; phenyl or substituted phenyl where the substituents
on the phenyl are from 1 to 3 of halogen, Cl-C6 aLkyl, Cl-C6 alkoxy or
hydroxy,
R14 and R15 are as defined above,
R4, R4a, R4b, R~ and R5a are independently hydrogen, phenyl, substl-
tuted phenyl, Cl-Clo alkyl, substituted Cl-Clo alkyl, where the
20 substituents on the alkyl or phenyl are from 1 to 5 of hydroxy, C1-C6
alkoxy, C3-C7 cycloalkyl, fluoro, Rl substituted or Rl, R2 indepen-
dently disubstituted phenyl C1-C3 alkoxy, Rl substituted or R1, R2
independently disubstituted phenyl, C1-C20-aLkanoyloxy, Cl-CS
alkoxycarbonyl, carboxy or formyl;
R4 and R5 can be taken together to form -(CH2)rB(CH2)S- where B is
CHRl, O, S(O)m or N-R10, r and s are independently 1 to 3 and R1 and
R10 are as defined above;
3 R6 is hydrogen, Cl -Clo alkyl or phenyl Cl -Cl o aLkyl;
WO 94/11012 PCr/US93/10551
~ 21~75~3
Ais
~8
--(CH2)x--C--(CH2)y--
. R8a
where x and y are independently 0-2;
R8, R8a and R8b are independently hydrogen, Cl-Clo alkyl, substituted
o C1-Clo alkyl where the substituents are from 1 to 3 of imidazolyl,
indolyl, hydroxy, fluoro, S(o)mR7a~ C1-C6 alkoxy, Rl, R2
independently disubstituted phenyl, C1-Cs-alkanoyloxy, C1-Cs
alkoxycarbonyl, carboxy, formyl, -NRlORl 1 where R10 and Rl 1 are
independently hydrogen, Cl-C6 aLkyl, or Cl-C5 alkanoyl-Cl-C6 alkyl;
or R8 and R8a can be taken together to form -(CH2)t- where t is 2 to 4;
and R8 and R8a can independently be joined to one or both of R4 and
R5 to form alkylene bridges between the terminal nitrogen and the alkyl
portion of the A group wherein the bridge contains from 1 to 5 carbon
atoms;
20 and pharmaceutically acceptable salts thereof.
Additional preferred compounds are realized in the above
structural formula when:
nisOor l;
25 pisOto3;
q is O to 2;
wisOor 1;
X is 0, S(O)m or -CH=CH-;
misOor 1;
Rl, R2, R1a, R2a, Rlb, and R2b are independently hydrogen, halogen,
C1-C7 alkyl, Cl-C3 perfluoroalkyl, -S(O)mR7a~ R7bo(cH2)v-
~R7bCoo(cH2)v-~ R7boco(cH2)v-~ phenyl or substituted phenyl where
the substituents are from 1 to 3 of halogen, Cl-C6 alkyl, Cl-C6 aLkoxy,
or hydroxy;
WO 94/1 1012 PCI /US93/10551
2~
- 10 -
R7a and R7b are independently hydrogen, C1-C6 alkyl, substituted Cl-
C6 alkyl, where the substituents are phenyl and v is O to 2;
5 R3a and R3b are independently hydrogen, R9, Cl-C6 alkyl substituted
with R9, phenyl substituted with R9, or phenoxy substituted with R9;
R9 is as defined above;
R12a, Rl2b and R12c are independently R5a or OR5a. R12a and R12b,
or R12b and Rl2c~ or R13 and Rl2b or Rl2a and R4b can be taken
together to form -(CH2)r-B-(CH2)S- where B is CHR1, O, S(O)m or
NR1 0, m is 0, 1 or 2, r and s are independently O to 2, Rl is as defined
above and R10 is hydrogen, Cl-c6 alkyl or Cl-cs alkanoyl-Cl-C6
5 aLkyl;
R13 is Cl-C6 alkyl, substituted Cl-C6 alkyl, where the substituents are
phenyl or sllbstituted phenyl phenyl or sllbstib~ted phenyl where the
substituents on the phenyl are from 1 to 3 of halogen, C1-C6 alkyl, Cl-
20 C6 alkoxy or hydroxy;
R14 and R15 are as defined above;
R4, R4a, R4b~ R5 and Rsa are independently hydrogen, C1-C1o alkyl,
25 substituted C1-C1o alkyl, where the substituents on the alkyl are from 1
to 5 of hydroxy, Cl-C6 alkoxy, fluoro, Rl substituted or Rl, R2
independently disubstituted phenyl, C1-C20-alkanoyloxy, C1-C5
alkoxycarbonyl or carboxy;
30 R6 is hydrogen or Cl-Clo alkyl;
WO 9 S/1 1012 PCI/US93/10551
~147~D~
A lS
~8
--(CH2)x--C--(CH2)y
R8a
where x and y are independently 0-2;
R8, R8a and R8b are independently hydrogen, C1-Clo alkyl, substituted
o C1-C10 alkyl where the substituents are from 1 to 3 of imidazolyl,
indolyl, hydroxy, fluoro, S(o)mR7a~ Cl-C6 alkoxy, Rl substituted or
Rl, R2 independently disubstituted phenyl, C1-C5-alkanoyloxy, C1-C5
alkoxycarbonyl, carboxy; or R8 and R8a can be taken together to form
-(CH2)t- where t is 2; or R8 and R8a can independently be joined to one
or both of R4 and R5 to form alkylene bridges between the tennin~l
nitrogen and the alkyl portion of the A group wherein the bridge
contains from 1 to 5 carbon atoms;
and pharmaceutically acceptable salts thereof.
Still further preferred compounds of the instant invention
are realized in the above structural formula when;
n is O or 1 ;
p is O to 2;
qis 1;
w is 1 ;
X is 0, S(O)m;
misOor 1;
R1, R2, Rla7 R2a~ Rlb~ and R2b are independently hydrogen, halogen~
Cl-C7 alkyl, Cl-C3 perfluoroalkyl, -S(O)mR7a, R7bo(cH2)v-~
R7bCoo(cH2)v-~ phenyl or substituted phenyl where the substituents
are from 1 to 3 of halogen, Cl-C6 aL~yl, Cl-C6 alkoxy, or hydroxy;
R7a and R7b are independently hydrogen, C1-C6 alkyl, substituted C1-
C6 alkyl, where the substituents are phenyl and v is O or 1;
WO 9~/t1012 PCI/US93/10551
- 12 -
R3a and R3b are independently hydrogen, R9, or C1-C6 alkyl
substituted with R9;
R9 is as defined above;
R12a, Rl2b and Rl2c are independently RSa. Rl2a and Rl2b~ or R12b
and R12C, or R13 and R12b or R12a and R4b can be taken together to
form -(CH2)r-B-(CH2)S- where B is CHRl, O, S(O)m or NR10, m is 0,
1 or 2, r and s are independently O to 2, R1 is as defined above and R10
is hydrogen, Cl-C6 alkyl or C1-Cs alkanoyl Cl-C6 alkyl;
R13 is C1-C6 alkyl, substituted C1-C6 alkyl, where the substituents are
phenyl or substituted phenyl; phenyl or substituted phenyl where the
substituents on the phenyl are from 1 to 3 of halogen, Cl-c6 alkyl, C1-
C6 alkoxy or hydroxy;
R14 and RlS are as defined above;
R4, R4a, R4b, R5 and RSa are independently hydrogen, C1-C1o alkyl,
substituted Cl-Clo alkyl, where the substituents on the alkyl are from 1
to 3 of hydroxy, Cl-C3 aLkoxy, fluoro, Rl substituted or Rl, R2 inde-
pendently disubstituted phenyl, Cl-C20 alkanoyloxy, C1-Cs alkoxy-
carbonyl or carboxy;
R6 is hydrogen;
Ais
~8
3 o --(CH2)x--Cl--(CH2)y
R8a
where x and y are independently 0-1:
WO 94/11012 PCI/US93/10551
~ 2I47~3
- 13 -
R8, R8a and R8b are independently hydrogen, C1-C1o alkyl, substituted
C1-C1o alkyl where the substituents are from 1 to 3 of imidazolyl,
indolyl, hydroxy, fluoro, S(o)mR7a~ Cl-C6 alkoxy, Rl substituted or
Rl, R2 independently disubstituted phenyl, Cl-C5-alkanoyloxy, C1-Cs
5 alkoxycarbonyl, carboxy; or R8 and R8a can be taken together to form
-(CH2)t- where t is 2; and R8 and R8a can independently be joined to
one or both of R4 and R5 to form aLkylene bridges between the terminal
nitrogen and the alkyl portion o~ the A group wherein the bridge
contains from 1 to 5 carbon atoms;
and pharmaceutically acceptable salts thereof.
Representative preferred growth hormone releasing
compounds of the present invention include the following:
1. (R)-a-[(3-Amino-3-methyl-1-oxobutyl)amino]-N-[[2'-(lH-
tetrazol-5-yl)[l ,1 '-biphenyl]-4-yl]methyl] -benzene-
b~lt~n~mide
2. (R)-oc-[(2-Amino-2-methyl-l-oxopropyl)amino]-N-[[2'-(lH-tetrazol-5-yl)[ 1,1 '-biphenyl]-4-yl]-methyl]benzene-
but~n~mide
3. (R)-o~-[[3-[2(R)-Hydroxypropyl]amino-3-methyl-1-oxo-
butyl]amino]-N-[[2'-(1H-tetrazol-5-yl)[1,1'-biphenyl]-4-
yl]methyl]benzenebut~n~mi(le
4. (R)-a-[[3-[2(S),3-Dihydroxypropyl]amino-3-methyl-1-
oxobutyl]amino]-N-[[2'-(1 H-tetrazol-5-yl)[ 1,1 '-biphenyl]-4-
yl]methyl]benzeneb--t~n~mitle
5 . (R)-oc-[(3-Amino-3-methyl- 1 -oxobutyl)amino] -N-[[2'-(1 H-
tetrazol-5-yl)[ 1,1 '-biphenyl]-4-yl]methyl]-benzenepentana-
mide
WO 9~/1101~ PCI/US93/~05~1
- 14 -
6. (R)-a-[(2-Amino-2-methyl- 1 -oxopropyl)amino] -N-[[2'-
(lH-tetrazol-5-yl)[ 1,1 '-biphenyl]-4-yl]-methyl]benzene-
pent~n~mide
7 . (R)-a-[[3-[2(R)-Hydroxypropyl]amino-3-methyl- 1-
oxobutyl]amino]-N-[[2'-( 1 H-tetrazol-5-yl)[ 1,1 '-biphenyl]-4-
yl]methyl]benzenepent~n~mide
8. (R)-a-[[3-[2(S),3-Dihydroxypropyl]amino-3-methyl-1-
o oxobutyl]amino]-N-[[2'-(lH-tetrazol-5-yl)[1,1'-biphenyl]-4-
yl]methyl]benzenepent~n~mide
9 . (R)-a-[(3-Amino-3 -methyl- 1 -oxobutyl)amino] -N-[[2'-( 1 H-
l 5 tetrazol-5-yl)[ 1,1 '-biphenyl]-4-yl]methyl]-lH-indole-3-
propanamide
10. (R)-a-[(2-Amino-2-methyl- 1 -oxopropyl)amino]-N-[[2'-
( 1 H-tetrazol-5-yl)[ 1,1 '-biphenyl] -4-yl] -methyl] -1 H-indole-
3-propanamide
1 1. (R)-a-[[3-[2(R)-Hydroxypropyl]amino-3-methyl-1-
oxobutyl]amino]-N-[[2'-(1 H-tetrazol -5-yl)[ 1, 1 '-biphenyl]-4-
yl]methyl]- 1 H-indole-3-propanamide
12. (R)-a-[[3-[2(S),3-Dihydroxypropyl]amino-3-methyl-1-oxo-
butyl]amino]-N-[[2'-(1 H-tetrazol-5-yl)[ 1,1 '-biphenyl] -4-
yl]methyl]- 1 H-indole-3-propanamide
13 . (R)-a-[(3 -Amino-3 -methyl- 1 -oxobutyl)amino] -N-[[2'-( 1 H-
tetrazol-5-yl)[ 1,1 '-biphenyl]-4-yl]me~hyl]-3-[(phenyl-
methyl)oxy]propanamide
WO 94/11012 PCI/US93/10551
21~7~
14. (R)-o~-[(2-Amino-2-methyl-1-oxopropyl)amino]-N-[[2'-
( 1 H-tetrazol-5-yl)[ 1,1 '-biphenyl]-4-yl] -methyl]-3-[(phenyl-
methyl)oxy]propanamide
15. (R)-a-[[3-[2(R)-Hydroxypropyl]amino-3-methyl-1-oxo-
butyl]amino]-N-[[2'-(1 H-tetrazol-5-yl)[ 1,1 '-biphenyl]-4-
yl]methyl] -3-[(phenylmethyl)oxy]-propanamide
16. (R)-a-[[3-[2(S),3-Dihydroxypropyl]amino-3-methyl-1-
oxobutyl]amino]-N-[[2'-(lH-tetrazol-S-yl)rl,l'-biphenyl]-4-
yl]methyl]-3-[(phenylmethyl)oxy] -propanamide
17 . (R)-a-[(3-Amino-3-methyl- 1 -oxobutyl)amino] -N-[[2'-(1 H-
tetrazol-5-yl)[ 1,1 '-biphenyl]-4-yl]-methyl]-3-[[(2,6-
difluorophenyl)methyl]oxy]-propanamide
18. (R)-a-[(2-Amino-2-methyl- 1 -oxopropyl)amino] -N-[[2'-
(1 H-tetrazol-5-yl)[ 1,1 '-biphenyl]-4-yl]-methyl] -3-[[(2,6-
difluorophenyl)methyl]oxy]-propanamide
19. (R)-a-[[3-[2(R)-Hydroxypropyl]amino-3-methyl-1-
oxobutyl]amino]-N-[[2'-(1 H-tetrazol-5-yl)[ 1,1 '-biphenyl]-4-
yl]methyl]-3-[[(2,6-difluorophenyl)-methyl]oxy]pro-
panamide
20. (R)-a-[[3-[2(S),3-Dihydroxypropyl]amino-3-methyl-1-
oxobutyl]amino]-N-[[2'-(lH-tetrazol-5-yl)[1,1 '-biphenyl]-4-
yl]methyl]-3-[[(2,6-difluorophenyl)-methyl]oxy]-
propanamide
21. (R)-4'-[[[2-[(3-Amino-3-methyl- 1 -oxobutyl)amino]- 1 -oxo-
4-phenylbutyl]amino]methyl]-N-ethyl[ 1,1 '-biphenyl]-2-
carboxamide
WO 91/11012 PCI/US93/105~1
2~
- 16 -
22. (R)-4'-[[[2-[(2-Amino-2-methyl- 1 -oxopropyl)-amino]- 1-
oxo-4-phenylbutyl]amino]methyl]-N- ethyl[ 1,1 '-biphenyl]-2-
carboxamide
23. (R)-4'-[[[2-[[3-[2(R)-Hydroxypropyl~amino-3-methyl-1-
oxobutyl]amino]- 1 -oxo-4-phenylbutyl]-amino]methyl] -N-
ethyl[ 1,1 '-biphenyl]-2-carboxamide
24. (R)-4'-[[[2-[[3-[2(S),3-Dihydroxypropyl]amino-3-methyl- 1-
o oxobutyl]amino]- 1 -oxo-4-phenylbutyl]-amino]methyl]-N-
ethyl[ 1,1 '-biphenyll-2-carboxamide
25 . (R)-4'-[[[2-[(3-Amino-3-methyl- 1 -oxobutyl)amino]- 1 -oxo-
l 5 5-phenylpentyl]amino]methyl]-N-ethyl[ 1,1 '-biphenyl]-2-
carboxamide
26. (R)-4'-[[[2-[(2-Amino-2-methyl- 1 -oxopropyl)-amino]- 1-
oxo-5-phenylpentyl]amino]methyl] -N-ethyl[ 1,1 '-biphenyl] -
2-carboxamide
27. (R)-4'-[[[2-[[3-[2(R)-Hydroxypropyl~amino-3-methyl-1-
oxobutyl]amino]- 1 -oxo-5-phenylpentyl]-amino]methyl]-N-
ethyl[ 1,1 '-biphenyl]-2-carboxamide
28. (R)-4'-[[[2-[[3-[2(S),3-Dihydroxypropyl]amino-3-methyl-1-
oxobutyl]amino]- 1 -oxo-5-phenylpentyl]-amino]methyl] -N-
ethyl[ 1,1 '-biphenyl]-2-carboxamide
29. (R)-4'-[[[2-[(3-Amino-3-methyl- 1 -oxobutyl)amino]- 1 -oxo-
3 3-(1 H-indole-3-yl)propyl]amino]methyl]-N-ethyl[ 1,1'-
biphenyl] -2-carboxamide
WO 94/1 1012 PCr/US93/10551
~7~
30. (R)-4'-[[[2-[(2-Amino-2-methyl- 1 -oxopropyl)-amino]- 1-
oxo-3-( 1 H-indole-3 -yl)propyl] amino] -methyl] -N-ethyl[ 1, 1 '-
biphenyl] -2-carboxamide
31. (R)-4'-[[[2-[[3-[2(R)-Hydroxypropyl]amino-3-methyl-1-
oxobutyl]amino]-l -o~o-3-(lH-indole-3-yl)propyl]amino]-
methyl]-N-ethyl[ 1,1 '-biphenyl]-2-carboxamide
32. (R)-4'-[[[2-[[3-[2(S),3-Dihydroxypropyl]amino-3-methyl-1-
o oxobutyl] amino] -1 -oxo-3 -(1 H-indole-3-yl)propyl] -
amino]methyl] -N-ethyl[ 1,1 '-biphenyl]-2-carboxamide
33 . (R)-4'-[[[2-[(3-Amino-3-methyl- 1 -oxobutyl)amino]- 1 -oxo-
3-[(phenylmethyl)oxy]propyl]amino]methyl]-N-ethyl[ 1, 1'-
1 5 biphenyl]-2-carboxamide
34. (R)-4'-[[[2-[(2-Amino-2-methyl-1-oxopropyl)-amino]-1-
oxo-3-[(phenylmethyl)oxy]propyl]amino] -methyl] -N-
ethyl[ 1,1 '-biphenyl]-2-carboxamide
35. (R)-4'-[[[2-[[3-[2(R)-Hydroxypropyl]amino-3-methyl-1-
oxobutyl]amino]- 1 -oxo-3-[(phenylmethyl)-oxy]propyl]-
amino]methyl] -N-ethyl [ 1,1 '-biphenyl] -2-carboxamide
36. (R)-4'-[[[2-r[3-[2(S),3-Dihydroxypropyl]amino-3-methyl-1-
oxobutyl]amino]- 1 -oxo-3-[(phenylmethyl)-oxy]propyl]-
amino]methyl]-N-ethyl[ 1,1 '-biphenyl] -2-carboxamide
37 . (R)-4'-[ [[2-[(3-Amino-3-methyl- 1 -oxobutyl)amino] -1 -oxo-
3 3-[[(2,6-difluorophenyl)methyl]oxy]propyl]-amino]methyl]-
N-ethyl[ 1,1 '-biphe~yl]-2-carboxamide
WO 94/l 1012 PCl/US93/~0551
- 18 -
38. (R)-4'-[[[2-[(2-Amino-2-methyl- 1 -oxopropyl)-amino]- 1-
oxo-3-[[(2,6-difluorophenyl)methyl]oxy]-propyl]amino] -
methyl] -N-ethyl[ 1,1 '-biphenyl] -2-carboxamide
39. (R)-4'-[[[2-[[3-[2(R)-Hydroxypropyl]arnino-3-methyl-1-
oxobutyl]amino]-1 -oxo-3-[[(2,6-difluoro-phenyl)methyl]-
oxy]propyl]amino]methyl]-N-ethyl-[ 1,1 '-biphenyl]-2-
carboxamide
o 40. (R)-4'-[[[2-[[3-[2(S),3-Dihydroxypropyl]amino-3-methyl-1-
oxobutyl]amino]-l -oxo-3-[[(2,6-difluoro-phenyl)methyl]-
oxy]propyl]amino]methyl]-N-ethyl-[ 1,1 '-biphenyl]-2-
carboxamide
41. (R)-a-[(3-Arnino-3-methyl-1-oxobutyl)arnino]-N-[[2'-
[[(methylamino)carbonyl]amino] [ 1,1 '-biphenyl]-4-
yl]me~yl]benzenebllt~n~mide
42. (R)-a-[(2-Amino-2-methyl- 1 -oxopropyl)amino]-N-[[2'-
2 o [ [(methylamino)carbonyl] amino] [ 1,1 '-biphenyl] -4-
yl]methyl]benzeneb-lt~n~mi~le
43. (R)-a-[[3-[2(R)-Hydroxypropyl]amino-3-methyl-1-
oxobutyl]amino]-N-[[2'-[[(methylamino)carbonyl]-
2 5 amino] [ l, l '-biphenyl] -4-yl]methyl]benzeneb lt~n~mi de
44. (R)-a-[[3-[2(S),3-Dihydroxypropyl]amino-3-methyl-1-
oxobutyl]amino]-N-[[2'-[[(methylamino)carbonyl]-
amino] [ 1,1 '-biphenyl]-4-yl]methyl]benzenebllt~n~mide
45 . (R)-a-[(3-Amino-3 -methyl- 1 -oxobutyl)amino] -N-[ [2'-
[[(methylamino)carbonyl]amino] [ 1,1 '-biphenyl]-4-
yl]methyl]benzenepent~n~mide
WO 9~/1 tO12 PCI/US93/10551
.
2~75
- 19 -
46. (R)-a-[(2-Amino-2-methyl- 1 -oxopropyl)amino]-N-[[2'-
[ [(methylamino)carbonyl] amino] [ 1 ,1 '-biphenyl] -4-
yl]methyl]benzenepent~n~mide
47. (R)-a-[[3-[2(R)-Hydroxypropyl]amino-3-methyl-1-
oxobutyl]amino]-N-[[2'-[[(methylamino)carbonyl]-
amino][l ,1 '-biphenyl]-4-yl]methyl]benzenepent~n~mide
48. (R)-a-[[3-[2(S),3-Dihydroxypropyl]amino-3-methyl-1-
o oxobutyl]amino]-N-[[2'-[[(methylamino)carbonyl]-
arnino] [ 1,1 '-biphenyl]-4-yl]methyl]benzenepent~n~mide
49. (R)-a-[(3-Amino-3-methyl-1-oxobutyl)amino]-N-[[2'-
[[( methylamino)carbonyl]amino][1 ,1 '-biphenyl]-4-
yl]methyl]-1 H-indole-3-propanamide
50. (R)-a-[(2-Amino-2-methyl- 1 -oxopropyl)amino] -N-[[2'-
[[(methylamino)carbonyl]amino][l ,1 '-biphenyl]-4-
yl]methyl]- 1 H-indole-3-propanamide
51. (R)-a-[[3-[2(R)-Hydroxypropyl]amino-3-methyl-1-
oxobutyl]amino]-N-[[2'-[[(methylamino)carbonyl]-
amino] [ 1,1 '-biphenyl]-4-yl]methyl] -1 H-indole-3 -
propanamide
52. (R)-a-[[3-[2(S),3-Dihydroxypropyl]amino-3-methyl-1-
oxobutyl]amino]-N-[[2'-[[(methylamino)carbonyl]-
amino][ 1,1 '-biphenyl]-4-yl]methyl]-1 H-indole-3-
propanamide
53. (R)-a-[(3-Amino-3-methyl-1-oxobutyl)amino]-N-[[2'-
[[(methylamino)carbonyl]amino] ~1,1 '-biphenyl]-4-
yl]methyl] -3-[(phenylmethyl)oxy] -propanamide
WO 94/ltO12 PCI/US93/10551
- 20 -
54. (R)-a-[(2-Amino-2-methyl- 1-oxopropyl)amino]-N-[[2'-
[[(methylamino)carbonyl]amino] [ 1,1 t-biphenyl]-4-
yl]methyl]-3-[(phenylmethyl)oxy] -propanamide
55. (R)-a-[[3-[2(R)-HydrOXypropyl]amillo-3-methyl- 1-
oxobutyl]amino] -N-[[2'-[[(methylamino)carbonyl]-amino] -
[ 1,1 '-biphenyl]-4-yl]methyl]-3-[(phenyl-methyl)oxy]propa-
namide
o 56. (R)-a-[[3-[2(S),3-Dihydroxypropyl]amino-3-methyl-1-
oxobutyl]amino]-N-[[2'-[[(methylamino)carbonyl]-
amino] [ 1,1 '-biphenyl]-4-yl]methyl]-3-[(phenyl-
methyl)oxy]propanamide
57. (R)-a-[(3-Amino-3-methyl-1-oxobutyl)amino]-N-[[2'-
[[(methylamino)carbonyl]amino] [ 1,1 '-biphenyl]-4-
yl]methyl]-3-[[(2,6-difluorophenyl)-methyl]oxy]-
propanamide
58. (R)-a-[(2-Amino-2-methyl-1-oxopropyl)amino]-N-[[2'-
[[(methylamino)carbonyl]amino] [ 1,1 '-biphenyl]-4-
yl]methyl]-3-[[(2,6-difluorophenyl)-methyl] oxy] -
propanamide
59. (R)-a-[[3-[2(R)-HydrOXypropyl]amillo-3-met
oxobutyl]amino]-N-[[2'-[[(methylamino)carbonyl]-
amino] [ 1,1 '-biphenyl] -4-yl]methyl] -3 -[ [(2,6-
difluorophenyl)methyl]oxy]propanamide
60. (R)-a-[r3-[2(S),3-Dihydroxypropyl]amino-3-methyl- 1-
oxobutyl]amino]-N-[[2'-[[(methylamino)carbonyl]-amino]-
[ 1,1 '-biphenyl]-4-yl]methyl] -3-[ [(2,6-difluorophenyl)-
methyl]oxy]propanamide
WO 94/1 1012 PCltUS93/10551
~ 2~ ~ 7~
61 . (R)-a-[(3-Amino-3 -methyl- 1 -oxobutyl)amino]-N-[ [2'-
hydroxymethyl[ 1,1 '-biphenyl] -4-yl]methyl]-benzene-
bllt~n~mide
62. (R)-a-[(2-Amino-2-methyl- 1 -oxopropyl)amino]-N-[[2'-
hydroxymethyl[ 1,1 '-biphenyl]-4-yl]methyl]-benzene-
bllt~n~mide
63. (R)-a-[[3-[2(R)-Hydroxypropyl]amino-3-methyl-1-
o oxobutyl]amino]-N-[[2'-hydroxymethyl[ 1,1 '-biphenyl]-4-
yl]methyl]benzenebllt~n~mide
64. (R)-a-[[3-[2(S),3-Dihydroxypropyl]amino-3-methyl-1-
oxobutyl]amino]-N-[[2'-hydroxymet,hyl[ 1,1 '-biphenyl]-4-
yl]methyl]benzenebllt~n~mide
65. (R)-a-[(3-Amino-3-methyl-1-oxobutyl)amino]-N-[[2'-
hydroxymethyl[l ,1 '-biphenyl]-4-yl]methyl]-benzene-
pent~n~mide
66. (R)-a-[(2-Amino-2-methyl- 1 -oxopropyl)amino]-N-[[2'-
hydroxymethyl[ 1,1 '-biphenyl]-4-yl]methyl]-benzene-
pent~n~mide
2 5 67. (R)-a-[[3-[2(R)-Hydroxypropyl]amino-3-methyl- 1-
oxobutyl]amino]-N-[[2'-hydroxymethyl[ 1,1 '-biphenyl]-4-
yl]met,hyl]benzenepent~n~mide
68 . (R)-a-[ [3-[2(S),3-Dihydroxypropyl]amino-3-methyl- 1-
3 oxobutyl]amino]-N-[[2'-hydroxymethyl[ 1,1 '-biphenyl]-4-
yl]methyl]benzenepent~n~mide
WO 94/11012 PCI/US93/105~1
- 22 -
69. (R)-a-[(3-Amino-3-methyl-1-oxobutyl)amino]-N-[[2'-
hydroxymethyl [ 1 ,1 '-biphenyl] -4-yl]mlethyl] -1 H-indole-3 -
propanamide
70. (R)-a-[(2-Amino-2-methyl- 1 -oxopropyl)amino]-N-[[2'-
hydroxymethyl[ 1,1 '-biphenyl]-4-yl]mlethyl]- 1 H-indole-3-
propanamide
71. (R)-a-[[3-[2(R)-Hydroxypropyl]amino-3-methyl- 1-
o oxobutyl]amino]-N-[[2'-hydroxymethyl[1,1'-biphenyl]-4-
yl]methyl]- 1 H-indole-3-propanamide
72. (R)-a-[[3-[2(S),3-Dihydroxypropyl]amino-3-methyl-1-
oxobutyl]amino]-N-[[2'-hydroxymethyl[ 1,1 '-biphenyl]-4-
yl]methyl]-1H-indole-3-propanamide
73. (R)-a-[(3-Amino-3-methyl-1-oxobutyl)amino]-N-[[2'-
hydroxymethyl[ 1,1 '-biphenyl]-4-yl]methyl]-3-[(phenyl-
methyl)oxy]propanamide
74. (R)-a-[(2-Amino-2-methyl- 1 -oxopropyl)amino]-N-[[2'-
hydroxymethyl[ 1,1'-biphenyl]-4-yl]m!ethyl]-3-[(phenyl-
methyl)oxy]propanamide
75. (R)-a-[[3-[2(R)-Hydroxypropyl]amino-3-methyl-1-
oxobutyl]amino]-N-[[2'-hydroxymethyl[ 1,1 '-biphenyl]-4-
yl]methyl] -3-[(phenylmethyl)oxy]propanamide
76. (R)-a-[[3-[2(S),3-Dihydroxypropyl]amino-3-methyl-1-
3 oxobutyl]amino]-N-[[2'-hydroxymethyl[ 1,1 '-biphenyl]-4-
yl]methyl]-3-[(phenylmethyl)oxy]propanamide
WO 94/1 1012 PCI /US93/105~1
- 23 -
77. (R)-a-[(3-Amino-3-methyl-1-oxobutyl)amino]-N-[[2'-
hydroxymethyl[ 1,1 '-biphenyl]-4-yl]methyl]-3-[[(2,6-
difluorophenyl)methyl]oxy]propanamide
78. (R)-a-[(2-Amino-2-methyl- 1 -oxopropyl)amino]-N-[[2'-
hydroxymethyl[ 1,1 '-biphenyl]-4-yl]methyl]-3-[[(2,6-
difluorophenyl)methyl]oxy]propanamide
79. (R)-a-[[3-[2(R)-Hydroxypropyl]amino-3-methyl- 1-
o oxobutyl]amino]-N-[[2'-hydroxymethyl[ 1,1 '-biphenyl]-4-
yl]methyl]-3-[[(2,6-difluorophenyl)-methyl]oxy]-
propanamide
80. (R)-a-[[3-[2(S),3-Dihydroxypropyl]amino-3-methyl-1-
oxobutyl]amino]-N-[[2'-hydroxymethyl[ 1,1 '-biphenyl]-4-
yl]methyl]-3-[[(2,6-difluorophenyl)-methyl]oxy]-
propanamide
81. (R)-a-[(3-Amino-3-methyl-1-oxobutyl)amino]-N-[[2'-
2 0 [[[(methylamino)carbonyl]amino]methyl]-[ 1,1 '-biphenyl]-4-
yl]methyl]benzeneb-lt~n~mide
82. (R)-a-[(2-Amino-2-methyl-1-oxopropyl)amino]-N-[[2'-
[[[(methylamino)carbonyl]amino]methyl]-[ 1,1 '-biphenyl]-4-
2 5 yl]methyl]benzenebllt~n~mi(le
83 . (R)-a-[[3-[2(R)-Hydroxypropyl]amino-3-methyl- 1-
oxobutyl]amino]-N-[[2'-[[[(methylamino)carbonyl]-
amino]methyl] [ 1,1 '-biphenyl] -4-yl]methyl]benzene-
3 bllt~n~mide
WO 9-1/11012 PCI/US93/~0551
- 24 -
84. (R)-a-[[3-[2(S),3-Dihydroxypropyl]amino-3-methyl-1-
oxobutyl]amino] -N-[[2'-[[[(methylamino)carbon-yl]-
amino]methyl] [ 1,1 '-biphenyl] -4-yl]methyl]-benzene-
bllt~n~mide
85. (R)-a-[(3-Amino-3-methyl-1-oxobutyl)amino]-N-[[2'-
[[[(methylamino)carbonyl]amino]methyl]-[ 1,1 '-biphenyl] -4-
yl]methyl]benzenepent~n~mide
o 86. (R)-a-[(2-Amino-2-methyl- 1 -oxopropyl)amino]-N-[~2'-
[[[(methylamino)carbonyl]amino]methyl]-[l ,1 '-biphenyl]-4-
yl]methyl]benzenepent~n~mide
87. (R)-a-[[3-[2(R)-Hydroxypropyl]amino-3-methyl-1-
oxobutyl]amino]-N-[[2'-[[[(methylamino)carbonyl]-
amino]methyl] [ 1,1 '-biphenyl] -4-yl]methyl]benzene-
pent~n~mide
88. (R)-a-[[3-[2(S),3-Dihydroxypropyl]~mino-3-methyl-1-
oxobutyl]amino]-N-[[2'-[[[(methylamino)carbon-
yl]amino]methyl][ 1,1 '-biphenyl]-4-yl~methyl]ben-
zenepent~n~mide
89. (R)-oc-[(3-Amino-3-methyl-1-oxobutyl)amino]-N-[[2'-
[[[(met,hylamino)carbonyl]amino]methyl]-[ 1,1 '-biphenyl]-4-
yl]methyl]- 1 H-indole-3-propanamide
90. (R)-a-[(2-Arnino-2-methyl- 1 -oxopropyl)amino] -N-[[2'-
[[[(methylamino)carbonyl]amino]methyl]-[ 1,1 '-biphenyl]-4-
yl]methyl]- 1 H-indole-3-propanamide
WO 94/11012 PCI/US93/10551
2 1 ~ 3
- 25 -
91. (R)-a-[[3-[2(R)-Hydroxypropyl]amino-3-methyl-1-
oxobutyl]amino]-N-[[2'-[[[(methylamino)carbonyl]-
amino]methyl][ 1 ,1 '-biphenyl] -4-yl]methyl] -1 H-indole-3-
propanamide
92. (R)-a-[[3-[2(S),3-Dihydroxypropyl]amino-3-methyl-1-
oxobutyl]amino]-N-[[2'-[[[(methylamino)carbon-yl]amino] -
methyl] [ 1,1 '-biphenyl] -4-yl]methyl]- 1 H-indole-3-
propanamide
93 . (R)-a-[(3-Amino-3-methyl- 1 -oxobutyl)amino]-N-[ [2'-
[[[(methylamino)carbonyl]amino]methyl]-[ 1,1 '-biphenyl]-4-
yl]methyl]-3-[(phenylmethyl)-oxy]propanamide
94. (R)-a-[(2-Amino-2-methyl- 1 -oxopropyl)amino] -N-[[2'-
[[[(methylamino)carbonyl]amino]methyl]-[ 1,1 '-biphenyl]-4-
yl]methyl]-3-[(phenylmethyl)-oxy]propanamide
95. (R)-a-[[3-[2(R)-Hydroxypropyl]amino-3-methyl-1-
oxobutyl]amino]-N-[[2'-[[[(methylamino)carbonyl]-
amino]methyl] [ 1,1 '-biphenyl]-4-yl]methyl]-3-
[(phenylmethyl)oxy]propanamide
96. (R)-a-[[3-[2(S),3-Dihydroxypropyl]amino-3-methyl-1-
oxobutyl]amino]-N-[[2'-[[[(methylamino)carbon-yl]-
amino]methyl] [ 1,1 '-biphenyl]-4-yl]methyl] -3-[(phenyl-
methyl)oxy]propanamide
97. (R)-a-[(3-Amino-3-methyl-1-oxobutyl)amino]-N-[[2'-
[[[(methylamino)carbonyl]amino]methyl]-[ 1,1 '-biphenyl]-4-
yl]methyl] -3-[[(2,6-difluoro-phenyl)-methyl]oxy]propana-
mide
WO 94/1 1012 PCr/US93/10551
- 26 -
98. (R)-o~-[(2-Amino-2-methyl- 1 -oxopropyl)amino]-N-[[2'-
[[[(methylamino)carbonyl]amino]methyl]-[ 1,1 '-biphenyl] -4-
yl]methyl]-3-[[(2,6-difluoro-phenyl)methyl]oxy]-
propanamide
99. (R)-oc-[[3-[2(R)-Hydroxypropyl]amino-3-methyl-1-
oxobutyl]amino]-N-[[2'-[[[(methylamino)carbonyl]-
amino]methyl] [ 1,1 '-biphenyl]-4-yl]methyl] -3-[[(2,6-
difluorophenyl)methyl]oxy]propanamide
100. (R)-o~-[[3-[2(S),3-Dihydroxypropyl]amino-3-methyl-1-
oxobutyl]amino]-N-[[2'-[[[(methylamino)carbon-
yl]amino]methyl] [ 1,1 '-biphenyl] -4-yl~methyl]-3-[[(2,6-
difluorophenyl)methyl]oxy]propanamide
Representative examples of the nomenclature employed are
given below:
CH
,O, ~ ~ 3
~CH2~NH CH2 NH2
HN o ,N=N
CH2~NH
WO 94/11012 PC~/US93/105~1
21~73~3
- 27 -
(R)-a-[(3-Amino-3-methyl- 1 -oxobutyl)amino]-N-[[2'-( 1 H-tetrazol-S-
yl)~ 1.1 '-biphenyll -4-vllmethyllbenzenepropanamide
o
~,H CH3' CH3
HN O O
CH2~NHCH3
(R)-a-[(2-Amino-2-methyl- 1 -oxopropyl)amino]-N-[[2'-[[(methyl-
amino)carbonyl~amino] [ 1 ,1 '-biphenyl]-4-yl]-methyl]-1H-imidazole-4-yl-
propanamide
~ o CH3~ ~CH3
CH2~NH 2 H H~.C~OH
HN O
CH2~¢~NHCH3
(R)-4'-[[[2-[[3-[2(R)-Hydroxypropyl~amino-3-methyl- 1 -oxobutyl3-
30 amino]-1-oxo-3-(lH-indole-3-yl)propyl~amino]-methyl~-N-methyl[l11'-
biphenyll -2-carboxamide
WO 9~/11012 PCI/US93/10551
~t~
- 28 -
,o, CH3~ ,CH3
C~ ,C~ N_CHz~ ,CH20H
[~3 CH2~S'CH2~ NH 2 H H~.C~oH
CH2~2oH
W
(S)-oc-[[3-[2(S),3-Dihydroxypropyl]amino-3-methyl-l -oxobutyl]amino] -
N-[[2'-hydroxymethyl[ 1, l '-biphenyl]-4-yl]methyl]-3-[(phenylmethyl)-
thiolpropanamide
The compounds of the instant invention all have at least one
5 asymmetric center as noted by the asterisk in the structural Formula I
above. Additional asymmetric centers may be present on the molecule
depending upon the nature of the various substituents on the molecule.
Each such asymmetric center will produce two optical isomers and it is
intended that all such optical isomers, as separated, pure or partially
20 purified optical isomers or racemic mixtures thereof, be included within
the ambit of the instant invention. In the case of the asymmetric center
represented by the asterisk in Formula I, it has been found that the
compound in which the 3-amino substituent is above the plane of the
structure, as seen in Formula Ia, is more active and thus more preferred
5 over the compound in which the 3-arnino substituent is below the plane
of the structure. This center will be designated according to the R/S
rules as either R or S depending upon the values of X, n, p and R15.
WO 94/11012 ~ PCl`/US93/10551
- 29 -
R15
I
(X)n
R3a~ (L)w--~C H2)q--,N - C - C--N, - C--A--N
The instant compounds are generally isolated in the form of
their ph~rm~ceutically acceptable acid addition salts, such as the salts
derived from using inorganic and organic acids. Examples of such
acids are hydrochloric, nitric, sulfuric, phosphoric, formic, acetic,
15 trifluoroacetic, propionic, maleic, succinic, malonic and the like. In
addition, certain compounds cont~inin~ an acidic function such as a
carboxy can be isolated in the form of their inorganic salt in which the
counterion can be selected from sodium, potassium, lithium, calcium,
m~nesium and the like, as well as from organic bases.
Compounds I of the present invention are prepared from
amino acid intermediates II as described in the following reaction
schemes.
Arnino acid intermediates 1 are, in some cases, commer-
cially available in the form of their N-t-butoxycarbonyl or N-
benzyloxycarbonyl derivatives. These intermediates can also be
prepared by a variety of methods described in the literature and
f~mili~r to one skilled in the art. For example, the Strecker synthesis
may be employed for the construction of racemic amino acid inter-
mediates. Resolution can be achieved by classical methods, for example
separation of diastereomeric salts by fractional cryst~lli7~tion. Alterna-
tively, a chiral amino acid synthesis may be employed using the
procedures described by R.M. Williams and M.N. Im (J. Amer. Chem.
Soc., 113, 9276-9286, 1991.). Conversion of the free amino acid
product to its N-t-butoxycarbonyl (BOC) derivative can be achieved by
a number of methods known in the art, for example, treatment with di-
WO 9~/t1012 PCI/US93/10551
2~5~
- 30 -
t-butyl dicarbonate in an inert solvent such as melhylene chloride.
Benzyloxycarbonyl (CBz) protected derivatives are obtained by
treatment of the amino acid with, for example, benzyl chloroformate.
SCHEME 1
Rl5 Rl 15
(Xl)n PhCH20H, EDC (Xl)n
O~ (C72)P DMAP, CH2,C12 (CHI2)p
,C--CH--H- BOC PhCH20- ,C,--CH--H- BOC
-- R15
CF3COOH, CH2C12 (Xl)n
or HCI, THF (c72)p
PhCH2O- ,C,--CH--NH2-HX
o
R15
2 5 3 aldehyde, I
sodium cyano- (Xl)n
borohydride (CH, 2)p
PhCH20- ,C, ~CH--~H2
R6
As shown in Scheme 1, formation of the benzyl ester 2 is
carried out by treatment with benzyl alcohol in the presence of a
coupling agent, such as 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide
hydrochloride (EDC), in methylene chloride with a catalytic amount of
WO 9~/1 1012 PCI'/US93/10551
21~7~
4-dimethylaminopyridine. Removal of the BOC protecting group
through the use of trifluoroacetic acid in methylene chloride or
hydrochloric acid in tetrahydrofuran gives the amine salt 3. Reductive
alkylation with an aldehyde and a mild reducing agent, such as sodium
5 cyanoborohydride, leads to the desired intermediate rl.
Attachment of the amino acid sidechain to intermediates of
formula II is accomplished by the route shown in Scheme 2. Coupling
is conveniently carried out by the use of an appropriately protected
amino acid derivative, such as that illustrated by formula III, and a
coupling reagent such as benzotriazol-l-yloxytris(dimethylamino)-
phosphonium hexafluorophosphate ("BOP") in an inert solvent such as
methylene chloride. Separation of unwanted side products, and
purification of intermediates is achieved by chromatography on silica
gel, employing flash chromatography (W.C. Still, M. Kahn and A.
5 Mitra, J. Org. Chem., 43, 2923 (1978)) or by medium pressure liquid
chromatography. Removal of the benzyl ester by hydrogenolysis or by
saponification in the presence of a strong base, such as sodium
hydroxide, affords the product IV. It may be appreciated by one skilled
in the art that the protecting group G must be selected to be compatible
20 with the conditions employed for removal of the specific class of ester
present in 4. Hence, as illustrated for the benzyl ester 4, G is taken as t-
butoxycarbonyl. It may further be appreciated that other combinations
of protecting group G and ester functionality may be employed; for
example, the benzyloxycarbonyl protecting group is inert to the
25 standard conditions of aqueous sodium hydroxide employed to
hydrolyze methyl or ethyl esters.
WO 94/11012 PCI/US93/105~1
.
S~3
- 32 -
SCHEME 2
R1s R, 4
(X)n HO--C--A--N--G
(CHl 2)p BOP, triethylamine
PhCH20- ,C,--CH--NH methylene chloride
R6
ll
IR15
(X)n H2, catalyst
(CHI2)p ,R4 NaOH, CH30H
PhCH20- ,C,--CH--N--C-A--N--G
R6
IR15
(X)n G is t-butoxycarbonyl
(CH2)p R4
"
HO-C--CH--N--C-A--N--G
R6
IV
The protected amino acid derivatives III are, in many cases,
3 0 commercially available in t-butoxycarbonyl (BOC) or benzyloxy-
carbonyl (CBz) forms. A useful method to prepare the preferred
sidechain 2 is shown in Scheme 3.
WO 94/1 1012 PCr/US93/10551
~ ~I47~3~-
- - 33 -
SCHEME 3
CH3
CH~C~ + O-C=NSO2Cl ether, ~ 3
CH3 CH3
6 NaH2 3~ ~CH3 BOC20 F¦ CH3
7 8
O CH3~ /CH3
8 LiOH~ HO,C--CH2--C--NHBOC
Reaction of isobutylene with N-chlorosulfonyl-isocyanate 5
in diethyl ether gives the azetidinone derivative 6. Removal of the
chlorosulfonyl group with aqueous sodium sulfite followed by reaction
with di-t-butyl-dicarbonate gives the BOC-protected intermediate 8.
Alk~line hydrolysis gives the protected amino acid derivative 2 in good
2 5 overall yield.
Attachment of the substituted phenyl sidechain V is
achieved as shown in Scheme 4. Using the aforementioned BOP
reagent, coupling is conveniently carried out in an inert solvent, such as
methylene chloride, to give compounds of formula VII in protected
3 o form.
W O 94/11012 PC~r/US93/105~1
- 34 -
S C~DE~EE 4
R15 NHR14
(X)n (C H2)q
(CH2)p 1 1 ~4
O ~6 ~/~
IV
V
~15
BOP, triethylamine (X)n
methylene chloride o(CH2)p R4
R14_N-C--CH--~--C--A--N-G
(CH2)q R6
(L)w
R1a i~ Vll
2~a/~JR3a
G is t-butoxycarbonyl or benzyloxycarbonyl
The substituted phenyl sidechains V are prepared from the
corresponding aLkylating agent VI by displacemel[lt of the leaving group
Y with sodium azide as shown in Scheme 5. Reduction of the azide
product by hydrogenation in the presence of a transition metal catalyst,
or alternatively by reaction with triphenylphosphine followed by
hydrolysis, gives the desired amine derivative 10. Conversion to the
desired intermediate V is achieved by the a~orementioned reductive
alkylation procedure.
WO 9~/1 1012 PCr/US93/10551
- 35 -
SCHEME 5
1. NaN3, DMF l H2
(C72)q 2. H2, catalyst (c72)q
(L) or (L)w
R2a~Rl3a Ph3P, H20 R2a~R3a
Vl 10
NH2 l HR14
(CH2)q aldehyde (C l z)q
sodlum J~
R2a~R3a cyanoborohydride R
V
Y is a leaving group
As illustratrated in Scheme 6 an alternative route involves
coupling of intermediate IV with R14NH2 using one of the coupling
reagents described previously, followed by aLkylatiQn of the amide bond
with VI. Alkylation is carried out in an inert solvent, such as
dimethylformamide, using a strong base such as sodium hydride or
potassium t-butoxide at temperatures of 0-100C. ALkylating agents VI
are, in some cases, commercially available or may be prepared by the
procedures described in the following schemes.
WO 94/11012 PCI/US93/105~1
- 36 -
SCHEME 6
Rl5
(X)n R14NH2, BOP
(CH2)p R4 triethylarnine
ll I methylene chloride
HO-C--CH--N-C--A--N-G
ll l
O R6
IV
IRl5
(X)n
(C72)p o ~4
R 14_ N {~ - CH--N--C--A--N--G
R15
IVa
1. NaH, DMF i)n
2 0 IVa 2. (C72)P o ~4
Y Rl4--N-C-CH--N-C--A--N-G
(C 1 2)q (C 1 2)q R6
~)w (l)W
Vl Vll
G is t-butoxycarbonyl or benzyloxycarbollyl
Alkylating agents VI are, in some cases cornmercially
available compounds or may be prepared by methods described in the
literature and familiar to one skilled in the art.
WO 94/11012 PCr/US93/10551
Compounds of formula I wherein R3a or R3b is a tetrazole
Cl.~.) are prepared as described in Scheme 7 by reaction of IV with a
suitably substituted intermediate 11 cont~ining a nitrile as tetrazole
precursor. Elaboration to the desired product 13 is carried out by
5 treatment with trimethyltin azide in boiling toluene.
SCHEME 7
NHRl4
~15 (CH2)q
HO-C--~CH--N--C -A--N--G R1 a ~
R6 R2a ~ CN
IV 11
BOP, triethylamine
methylene chloride
WO 9~/1 1012 PCr/US93/10551
.
q ~
- 38 -
SCHEME 7 (cont'd)
R15
I
(X)n
(C72)P R IR4
R14_N--C--CH--N--C -A--N--G
(CH2)q
(L)w (CH3)3snN3
~J~ toluene, heat
R1a ' --CN
n2a
n 12
R15
(X)n
0( 72)p R4
Il 11 1
R14--N--C--CH--N--C--A--N--G
2 0 (CH2)q R6
(L)w
2 ~/~ N-N
13
G=t-butoxycarbonyl or benzyloxycarbonyl
WO 94/11012 PCI/US93/}0551
~ 7~D3
- 39 -
A useful method to prepare the preferred intermediate 18
is shown in Scheme 8, and in U.S. Patent 5,039,814.
SCHEME 8
N=N
~ NaN3, ZnCI2 ~N,NH
14
N=N
Ph3CCI, Et3N ~ ,N-CPh3
CH3CN
1. nBuLi
2. ZnCI2
ClZn N= N
~f ~ ,N--CPh3
2s 16
WO 9~/11012 PCr/US93/105~1
4~
- 40 -
SCHEME 8 (cont'd)
CH3
,N-CPh3 ¢~
16
17 Ni(PPh3)2Cl2
CH2Br
N-bromosuccinimide ¢~ N=N
AIBN ~ ,N--CPh3
18
2~
As outlined in Scheme 8, benzonitrile is treated with
sodium azide and zinc chloride to give 5-phenyltetrazole 14 which is
converted to the N-trityl derivative 15 by treatment with triphenyl-
methyl chloride and triethyl~mine. The zinc reagent 16 was prepared
25 by treatment with n-butyl lithium followed by zinc chloride. Coupling
with 4-iodotoluene using the catalyst bis(triphenylphosphine)nickel(II)
dichloride gives the biphenyl product 17 in high yield. Reaction with
N-bromosuccin~mide and AIBN gives bromide 18. Conversion to the
requisite amine derivative V is achieved by the procedure described in
30 Scheme 5.
Compounds of Formula I wherein R3a or R3b are taken as
R4RSNco can be prepared by several methods. For example, as shown
in Scheme 9, compound 20 wherein R4 and R5 are both hydrogen is
conveniently prepared by hydrolysis of a nitrile precursor 19.
WO 94/11012 PCI`/US93/105~1
~ 1 ~ 7 ~ ~ 3
- 41 -
SCHEME 9
R15
I
(X)n
o (CH2)p R4
R14--N--C--CH--IN--C-A--N--G
(CHI 2)q R16
(L)w K2C03, H22
0 ~ DMSO
`1
/~
R2a
19
R15
(X)n
o(CHI2)p R, 4
R14--N--C--CH--I--C-A--N--G
(CHI 2)q R16
(L)w
R1a~ CN--NH2
R2a
3o Thus, treatment of the nitrile 19 with hydrogen peroxide
and a strong base, such as potassium carbonate, in a polar solvent, such
as dimethylsulfoxide at temperatures of 25C to 150C results in
formation of the amide derivative 20.
A useful method of preparing the intermediate 23 is
outlined in Scheme 10.
WO 94/11012 PCI/US93/10551
.
,4.q;~g`
- 42 -
SCHEME 10
CH3 Br
~ CN Pd(pph3)2cl2
+ ~ DMF, 100C
Sn(CH3)3
21
CH3 CH2Br
NBS 1~1
¢~CN AIBN ~CN
22 23
Thus, treatment of 4-(methylphenyl)l:rimethyl st~nn~ne 21
with 2-bromobenzonitrile in dimethylformamide at 100C in the
presence of bis-triphenylphosphine palladium (II) chloride results in
2 coupling to forrn the biphenyl nitrile 22 in high yield. Conversion to
bromide 23 is achieved by treatment with N-bromosuccinimide and a
radical initiator, such as azobisisobutyronitrile (AIBN), in refluxing
carbon tetrachloride. Conversion to the requisite amine derivative V is
achieved by the procedure described in Scheme 5.
Compounds of Formula I wherein R3a or R3b are taken as
R4R5NCo and R4 and/or R5 are other than hydrogen are prepared
from the corresponding carboxylic acid derivatives 24 as shown in
Scheme 11 .
WO 9~/11012 2 PCI/US93/10551
- 43 -
SCHEME 1 1
- R15
(X)n
o (CHl 2)p R4
R14--N--C--CH--N--C-A--N--G
(CH2)q R6
(L)w R4R5NH
~ BOP
~J CH2cl2
R2a
IR15
(X) n
2 0 R (C I 2)p 8 R, 4
R14--N--C--CH--N--C-A--N--G
(CH, 2)q R6
(L)W
2 5 R 1 a--~--CN--N R4R5
R2a
Coupling of the carboxylic acid derivative 24 with
R4R5NH is conveniently carried out by the use of a coupling reagent
such as benzotriazol-l-yloxytris-(dimethylamino)phosphoniurn
hexafluorophosphate ("BOP") in an inert solvent such as methylene
chloride.
WO 9~/11012 PCltUS93/10551
.
l ~ Q s3
- 44 -
The requisite carboxylic acid precursors can be prepared as
illustrated in Scheme 12 for the biphenyl compowld 24.
WO 94/11012 PCI/US93/10551
7 ~ ~ ~
- 45 -
SCHFMF 12
R15 CH2NH2
(X~n ~
5 (CHI2)p 1l R4 + ~
HO- IC--CH--N-C-A-N-G ~,COOtBu
O R6
IV
IR15
(X)n
BOP,triethylamine ~~ 7 P R IR4
R14--N-C--CH--N-C-A-N-G
methylene chlonde l I
CH2 R6
¢~
2 o ¢~cootBu
27 G is benzyloxycarbonyl
Rl 15
(X)n
CF3COOH o(CH2)p R4
CH2C12 R14--N- C--CH--N- C-A -N- G
(CH2)q R6
[~
~,COOH
24 ~
WO 94/11012 PCr/US93/105~1
.
- 46 -
A convenient method to prepare the useful intelmediate 30
is shown in Scheme 13. Metallation of 4-iodotoluene with t-butyl-
lithium in tetrahydro-furan, followed by treatment with zinc chloride
gives the intermediate zinc reagent 28. Coupling of 28 with t-butyl 2-
5 bromobenzoate in the presence of bis(triphenylphosphine)nickel(II)chloride affords the biphenyl product 29 in high yield. Bromination to
give the desired intermediate 30 is carried out under the aforementioned
conditions. Conversion to the requisite amine derivative V is achieved
by the procedure described in Scheme 5.
SCHEME 13
CH3 CH3 Br
1. 2 t-BuLi, THF, -78C ~ ~CO2tBu
2. ZnCI2, ether ~
ZnCI _ Ni(PPh3)2CI2
28
CH3 CH2Br
¢~ N-bromosuccinimide
CCI4, AIBN, heat
2 5 ~CO2tBU ~CO2tBu
29 30
Compounds of formula I where R3a or R3b is a carbamate.
semicarbazide or urea derivative, wherein this functionality is attached
to the phenyl ring by a nitrogen atom are prepared from intermediate
32, obtained by reaction with a derivative 31 wherein R3a or R3b is a
nitro ~roup as shown in Scheme 14.
WO 94/1 1012 PCT/US93/~0551
- 47 -
SCHEME 14
NHRl4
R15 (cHl2)q
(X)n t
(c72)p 8 IR R1a~
HO- ,C,--CH--IN--C--A--N-G + 2/`~ N2
R6
IV
R15
1 5 (X)n
lo~ (C72)p 0 ~4
BOP, triethylamine R14_N-C--CH--N-C-A-N-G
methylene chloride (CHI 2)q R6
2 0 (L)w
R1a~
R2a NO2
2S 32
G is t-butoxycarbonyl
A useful method of synthesizing a preferred intermediate
36 is shown in reaction Scheme 15.
WO 94/t 1012 PCI/US93/10551
- 48 -
SCHEME 15
~ NO2 PdgPh3)4, NaO~
2-propanol,
B(OH)2 benzene heat 1~1
o ~3 34
CH2Br
1 5 N-bromosuccinimide
CCI4, AIBN, heat
0, NO2
36
Reaction of 4-tolylboronic acid 33 w~.th 2-bromo-
nitroben~ene 34 in the presence of a transition mel-al catalyst such as
25 (tetrakis)triphenylphosphine p~ m (O) in a mi.xed solvent system
cont~ining aqueous sodium hydroxide, water, 2-propanol and benzene at
elevated temperatures for several hours gives the coupled product 35 in
good overall yield. Chromatographic purification and separation of
unwanted by-products is conveniently performed on silica, eluting with
3 0 common organic solvents such as hexane, ethyl acetate and methylene
chloride. Conversion of 3~ to the bromide derivative 36 is
accomplished by treatment with N-bromosuccinimide in refluxing
carbon tetrachIoride in the presence of a radical initiator such as
benzoyl peroxide or 2,2-azobisisobutyronitrile (A:rl3N). Conversion to
WO 94tl1012 PCI/US93/105~1
~ 21 ~ 7~ ~3
- 49 -
the requisite amine derivative V is acheived by the procedure described
in Scheme 5.
As shown in Scheme 16, reduction of the nitro group of 32
is achieved by hydrogenation in the presence of a metal catalyst, such as
palladium on carbon, in a protic solvent such as methanol or ethanol. It
may be appreciated by one skilled in the art that for certain compounds
where catalytic hydrogenation is incompatible with existing
functionality, alternative methods of reduction are indicated, such as
chemical reduction with stannous chloride under acidic conditions. It
should also be noted that the protecting group G in intermediate 32 must
be compatible with the experimental conditions anticipated for reduct-
ion. For example, intermediate 32 wherein G is t-butoxycarbonyl
(BOC) are stable to the conditions of catalytic reduction employed in the
conversion to ~7. Intermediate 37 may also be further elaborated to
15 new intermediate 38 by reductive aLt~ylation with an aldehyde by the
aforementioned procedures.
WO 94/11012 PCr/US93/105~1
rl5~
- so -
SCHEME 16
Rl5
I
(X)n
o (CH2)p IR4
R14_N-C--CH--N-C--A--N-G
(CHI 2)q R6
(L)w H2, PcVC
o Rla~ CH30H
N0 32
R16
(X)n
o (CH2)p IR4
ll l ll
R14_N-C--CH--N-C--A--N-G
(CHI 2)q R6
R1a ' J
2/~ NH
37 G is t-butoxycarbonyl
wo g~ 012 21 ~ 7~ ~ 3 PCr/l iS93/10551
SCHEME 16 (cont'd)
F~15
(X)n
O (CH2)p o ~4
R14_N-C--CH--IN-C--A--N-G
(CHI 2)q R6
aldehyde
R1a_ ] sodium cyanoborohydride
37
F~15
(X)n
20 R (CI H2)p 0 ~4
R14_N-C--CH--N-C--A--N-G
(CHl2)q R6
(L)w
2 5f~ NH R12b
38 G is t-butoxycarbonyl
Elaboration of 37 to carbamate compounds is achieved by
reaction with the appropriate chloroformate reagent in pyridine or in
methylene chloride with triethyl~mine as shown in Scheme 17.
WO 9~/11012 PCI/US93/10551
t~3 - 52 -
SCHEME 17
~7~15
(X)n
,O, (7H2)p ,O, ~4
R14--N-C--CH--N-C--A--N-G
(CHI 2)q R6
(IL)w R13-~
o R1a~ pyridine or
R2a NH2 methylene chloride/Et3N
37
~15
(X)n
R (7H2)P R ~4
R14_N-C--CH--N-C--A--N-G
(CHI 2)q R6
(L)W
R~ ~R13
Transformation of amine intermediate 37 to urea
derivatives is accomplished in several ways. Terrnin~lly disubstituted
compounds 40 can be obtained directly by reaction of 37 with a
disubstituted carbamoyl chloride in an inert solvent such as methylene
chloride in the presence of triethylamine or 4-dimethylaminopyridine.
WO94/11012 ~ 3 4 7~D3 PCI/US93/~0551
.
- 53 -
In addition, monosubstituted compounds 41 wherein either R4b or R12a
is hydrogen are obtained from 37 by reaction with an isocyanate as
shown in Scheme 18.
SCHEME 18
~15
(X)n
1 o o (CH2)p R lR4
R14_N-C--CH--N-C--A--N-G
(c72)q R6 ~4b
~w Rl2a~
R~a ~ NH2 methylene chloride/Et3N
4-dimethylaminopyridine
R15
(X)n
R (CH2)p R ~4
R14_N-C--CH--I -C--A--N-G
2S (CH2)q R6
(L)W
R1a~ 4b
3 0 ~ R
WO 94/1101~ PCI`/US93/10551
?~ 4~ ~ 54 -
SCHEME 18 (cont'd)
F~15
(X)n
R (CIH2)p R 1~4
R14_N-C--CH--N-C--A--N-G
(CHI 2)q R6
1 o (L)w
R1a--~ R12a-N=C=O r
2~/~\NH rnethylene chloride/Et3N
4-dimethylaminopyridine
37
~15
(X)n
R (CIH2)p R ~4
R14_N-C--CH--N-C--A--N-G
(CHI 2)q R6
(L)W
~2~ R 1 2a
Alternatively, amine 37 is converted to an isocyanate 42 by
treatment with phosgene or an equivalent reagent such as bis(trichloro-
methyl)carbonate (triphosgene) as indicated in Scheme 19. Subsequent
reaction of 42 with primary or secondary amines in an inert solvent
WO 94/ltO12 PCI/US93/105~1
2117~
such as methylene chloride gives the corresponding urea derivates 43 in
good yield. Isocyanate 42 is also converted to substituted semi-
carbazides 44 or hydroxy- or alkoxyureas 45 by reaction with
substituted hydrazines or hydroxy- or alkoxylamines, respectively.
SCHEME 19
R15
I
(X)n
,0 (Cl H2)p o ~4
R1 _N-C--CH--IN-C--A--N-G
(CH2)q R6
(L)w
R1a~ ~ (Cl3c0)2Co
J methylene chloride
R2a NH2 triethylamine
~15
(X)n
14 ~0, (CH2)p 0 ~4
R --N-C--CH--I -C--A--N-G
(CHI 2)q R6
(L)W
R2a~N=C=O G is t-butoxycarbonyl
or
42 benzyloxycarbonyl
WO 9~/11012 PCI/US93/10551
~ 4~ 56 -
SCHEME 19 (cont'd)
~15
(X)n
,0l (CI H2)P R F'~4
R14_N-C--CH--N-C--A--N-G
(CHI 2)q R6
1 0 (L)w
R1a~ R4bR12aNH
2a ~ N=C=O methylene chloride
R
42
F~15
(X)n
R (CIH2)p R F~4
R14_N-C--CH--N-C--A--N-G
(CHI 2)q R6
2 5 (L~w
R2a~lNH~N` R12a G is t-butoxycarbonyl
O or
3 0 43 benzyloxycarbonyl
wos4/ll0l2 21~7- ~3 PCI/US93/10551
SCHEME 19 (cont'd)
~15
(Xl)n
,o, (CI H2)P R ~4
R14_N-C--CH--N-C--A--N-G
(C72)q R6
(L)w
R1a~ R4bR12aNN(R12b)H
J methylenechloride
42
~15
(xl)n
,o, (Cl H2)p 0 ~4
R14_N-C--CH--N-C--A--N-G
(CHI 2)q R6
(L)w
~ H~f N 4b G is t-butoxycarbonyl
44 benzyloxycarbonyl
WO 91/11012 PCI/US93/105~1
- 58 -
SCHEME 19 (cont'd)
7~15
(X)n
,o (Cl H2)p O ~4
R14_N-C--CH--N-C--A--N-G
(CHI 2)q R6
(L)w
R1a~ R4b(R5ao)NH
2~/~\NH methylene chloride
42
~15
(X)n
o (Cl H2)p O ~4
R14_N-C--CH--N-C--A--N-G
(CHI 2)q R6
(L)w
R1a~ ol R5a
R2a HN~N~ R4b G is t-butoxycarbonyl
O or
benzyloxycarbonyl
Compounds of formula I where R3a or R3b is a carbazate
or carbamate derivative where attachment to the phenyl ring is through
the oxygen atom of the carbazate or carbamate linkage are prepared
from acetophenone intermediates 46 as indicated in Scheme 20.
~0 9~/11012 PCI/US93t10551
- 59 -
SCHEME 20
~15
(X)n
,01 (7H2)P 1~4
R14_N-C--CH--N-C--A--N-G
(cHl2)q R6
(L)w
Rla--~I m-chloroperbenzoic acid
methylene chloride
R2a o~ 3
46
~15
(xl)n
20,O, (Cl H2)p o ~4
R14_N-C--CH--N-C--A--N-G
(CHI 2)q R6
(L)w
~O~cH
WO 94/11012 PCr/US93/10551
~3~
- 60 -
SCHEME 20 (cont'd)
~15
(X)n
R (CI H2)P R ~4
R --N-C--CH--N-C--A--N-G
(c72)q R6
(L)w
R1a~q R12a N=C=O
2~/~\;OH methylene chloride/Et3N
4-dimethylaminopyridine
LiOH48
1 5 H20
47 R15
I
(X)n
20 o (Cl H2)p 0 ~4
R14_N-C--CH--N-C--A--N-G
(c72)q R6
(L)w
R1a--~1
R2a/~ ~N`R12a
49 o G ist-butoxycarbonyl
benzyloxycarbonyl
WO94/11012 2I ~ 7~ ~3 PCI'/US93/105~1
- 61 -
SCHEME 20 (cont'd)
R1~
I
(X)n
o (Cl H2~P ~~ ,R4
R14_N--C--CH--N--C-A--N--G
(CHI 2)q R6
(L)W
R1a~ 1. N, N'-carbonyldiimidazole
/~ 2. R4bR12aNN(R12b~H
R2a OH methylenechloride
48
lS
Rl 15
(xl)n
2~ ~o~ (Cl H2~P ,R4
R14_N--C--CH--N--C-A--N--G
(CHl 2)q R6
(L)W
2 S R 1 a~ R 1 Zb
O G is t-butoxycarbonyl
benzyloxycarbonyl
Oxidative rearrangement of 46 through the use of a
peroxy-carboxylic acid (Baeyer-Villager reaction) such as m-
chloroperbenzoic acid gives the ester 47 which is hydrolyzed in the
presence of a strong base such as sodium or lithium hydroxide to give
phenol 48.
WO 94/11012 PCI/US93/105~1
.
- 62 -
Reaction of 48 with an isocyanate leads directly to
carbamate analogs 49. Additionally, treatment of 48 with N,N'-
carbonyldiimidazole in dimethylformamide can form an activated
intermediate which will react with substituted hydrazine reagents to give
5 carbazate products 50.
Compounds of formula I wherein R3a or R3b contains the
linkage -CH2N(R12b)- can be prepared from the t-butyl ester inter-
mediate 51 as described in Scheme 21. Removal of the t-butyl ester
through the use of trifluoroacetic acid gives the carboxylic acid 24. It
may be appreciated by one skilled in the art that the protecting group G
in 51 must therefore be compatible with the strongly acidic conditions
employed for ester cleavage, hence G is taken as benzyloxycarbonyl.
Conversion of the carboxylic acid to the benzyl~mine derivative 52 can
be achieved by a five-step sequence consisting of: 1) formation of a
15 mixed anhydride with isobutyl chloroformate; 2) reduction with sodium
borohydride to the benzyl alcohol; 3) formation of the mesylate with
methanesulfonyl chloride; 4) formation of the azide by reaction with
sodium azide, and finally, 5) reduction of the azide with tin(II) chloride.
The benzylamine intermediate 52 can be further elaborated to 53 by the
20 aforementioned reductive ~min~tion procedure.
PCI /US93/10551
WO 94/t 1012
2~7~
- 63 -
SCHEME 21
F~15
(I)n
R(CIH2)p O r4
R14--I -C-CH--IN--C--A--N-G
D6
(CH2)q 1~
(L)w
R1a~ ~ COOtBu
R2a CF3COOH
51 methylene chloride
Flt15
(I )n
R ( IcH2)p R ,R4
R14--IN-C-CH--Nl--C--A--N-G
(CH2)q R
(L)w
R 1 a --C"
l/~J OH
D2a
n
24 G is benzyloxycarbonyl
WO 9~/ 11 0 1 2 PCrt US93/ 1 055 1
- 64 -
SCHEME 21 (Cont'd)
1. iBu-O-C-CI, N-methyl-
morpholine
24 2. sodium borohydride
.
3. CH3SO2CI
4. sodium azide
5. SnCI2, aqueous dioxane
R15
I
(I )n
O (CH2)p R4
R14--N--C--CH--N--C--A--N--G
(C 112)q R 6
(L~W
Rla ~J--CH2NH2
R2a
52 G is benzyloxycarbonyl
2s
WO 94/1 1012 PCr/US93/105~1
21~7~
- 65 -
SCHEME 21 (Cont'd)
~15
. (I)n
R (CI H2)p o ~4
R14--IN-C-CH--N--C--A--N-G
(C lH2)q
(L)w
R1a~ CH2NH2
R2a aldehyde
5? sodium
cyanoborohydride
IRl5
2 0 ( I )n
O (CH2)p R4
R14--N-C-CH--N--C--A--N-G
(CH2)q R6
(L)W
R1a ~ CH2NHR12b
R2a
~ G is benzyloxycarbonyl
WO 94/tlO12 PCI/US93/105~1
. .
~ ~ 4~ ~
- 66 -
Reaction of amine 53 with the appropriate reagents to form
urea-linked compounds 54 and 55 carbamate-linked compounds 56, and
amide-linked structures 57 are illustrated in Scheme 22.
WO 91/11012 PCI/US93/10551
~ 67 2I47~
G--Z G
~ I-Z
0=0 ~ ~0
--Z
~'I -- N
tr x~
O=~
Z~
G G
G
A
,~
O o
V~ _
~ al
G- I G
":
O= C~
~D G
0 Z~
U~ c C`J ~
G--X--c )--c~
O=
~_N_3
~t -- =/
C~
tr
WO 94/11012 PCr/US93/105~1
68
~ I ~
~--Z
C~l I
Z
O = ~ O
CL z--G G--z
G X--C)--C)
G ~ G
~ G
Z
O a)
C`l o
~I 0~
~I~ N I C)
G--Z
~ I Z
~t I G ~ LLI
~--Z
O= ( ) D
CD ~
a:--X--c~--c~ I
=~ I 3~ ~ N
r
G
~VO 9~/ 1 1 0 1 2 6~ ~ 4 7 ~ ~ r Pcr/ US93/ 1 055 1
C~
1 ~ ~ Z t~
O
O = (.) ~C O
Q Z-G a-z
L C I
~:--X~
O=~ ~
Z--I_~ ~ ~ ~¦
~,,1 _ ~ =,,\
G G
z
~ ~ a) c~
~--Z
O=
Q Z--G I
tr: X--C~
O=
r c
G (~
WO 9~/11012 PCI/US93/10551
?~ ~ 4~ ~
G--lZ ~D
~ G
O=c~ ~ ~0
<L~ G- Z
Q Z-G
' I = I
G--X--C~
G ~c
Z
~L ~
.L O Z -)
~ O=C~
v~ I Q-.
(~ G m
G--Z
~t
O = ~ N
Q ~- G
G X--C )--( )
o=~
C~l
G (~ .
WO 94/11012 PCI/US93/105~1
~ 21~7~1~3
A useful preparation of the protected benzylamine
intermediate 62 is shown in Scheme 23. Metallation of 4-bromobenzyl
t-butyldimethylsilylether 5~ with n-butyllithium followed by treatment
5 with trimethyl borate gives the aryl boronic acid 59. Reaction of 59
with 2-bromo-N-(t-butoxycarbonyl)benzylamine 60 in the presence of
tetrakis(triphenylphosphine)palladium(0) and barium hydroxide in
aqueous 1,2-dimethoxyethane at elevated temperature gives the coupled
product 61 in good yield. Desilylation is carried out by treatment with
tetra-n-butylammonium fluoride; conversion to the O-methanesulfonate
62 is achieved by reaction of the intermediate benzyl alcohol with
methanesulfonic anhydride. Conversion to the requisite amine
derivative V is achieved by the procedure described in Scheme 5.
SCHE~IE 23
CH20Si(CH3)2t-Bu CH20Si(CH3)2t-Bu
~ 1. n-C4H9Li, ¢~
Br THF B(OH)2
2. B(OGH3)3
58 59
Br di-t-butyl- Br
,~CH2NH2 dicarbonate ~,CH2NHBOC
3 0 ~ chloride W
WO 94/11012 PCr/US93/105~1
2~ 72-
SCHEME 23 (Cont'd)
Br CH20Si(CH3)2t-Bu
~CH2NHBOC
B(OH)2
~iQ 59
1 o CH20Si(CH3)2tBu
Pd(PPh3)4
Ba(OH)2 ~
aqueous dimethoxy- 1 CH2NHBOC
ethane, heat ~ ~
CH20Si(CH3)2tBu ~
ll l
~ 1. (n-c4Hs)4NF
~ CH2NHBOC 2. (CH3S02)20,
(c2H~)3N
61
2 5 C H2OS02CH3
BOC is t-butoxycarbonyl
~,CH2NHBOC
30 ~
61
WO 94/1 1012 PCI/US93/10551
~ 21~7~0~
Conversion to the final products of formula I wherein R5 is
hydrogen, is carried out by simlllt~neous or sequential removal of all
protecting groups from intermediate VII as illustrated in Scheme 24.
Removal of benzyloxycarbonyl groups can be achieved by a number of
5 methods known in the art; for example, catalytic hydrogenation with
hydrogen in the presence of a pl~tinum or palladium catalyst in a protic
solvent such as methanol. In cases where catalytic hydrogenation is
contraindicated by the presence of other potentially reactive
functionality, removal of benzyloxycarbonyl groups can also be
achieved by treatment with a solution of hydrogen bromide in acetic
acid. Catalytic hydrogenation is also employed in the removal of N-
triphenylmethyl (trityl) protecting groups. Removal of t-butoxy-
carbonyl (BOC) protecting groups is carried out by treatment of a
solution in a solvent such as methylene chloride or methanol, with a
strong acid, such as hydrochloric acid or trifluoroacetic acid.
Conditions required to remove other protecting groups which may be
present can be found in Protective Groups in Organic Synthesis.
WO 94/11012 PCI/US93t105~1
- 74 -
SCHEME 24
IRl5
( I )n
O (CH2)p R4
R14--N--C-CH--IN--C-A--N--(,
(CH2)q R6
(L)w
R3a Removal of
protecting groups
J
R1a R2a
Vll
~:~15
( I )n
R (CI H2)P R F~4
R14--N--C - CH--N--C -A--N--G
(CH2)q R6
(L)w
R1a
l/~\
3 0 R2a R3a
Compounds of formula I wherein R4 and R5 are each
hydrogen can be further elaborated by reductive alkylation with an
WO 94/11012 PCI/US93/10551
~1~7~
aldehyde by the aforementioned procedures or by alkylations such as by
reaction with various epoxides as shown in Scheme 25. The products,
obtained as hydrochloride or trifluoroacetate salts, are conveniently
purified by reverse phase high performance liquid chromatography
5 (HPLC) or by recrystallization.
SCHEME 25
R15
( lx)n
o (CH2)p R4
R14--N -C- CH--N--C--A--N -H
R6
(C ~2)q
(-)w Aldehyde,
sodium cyanoborohydride
R1a~ or epoxides
2 o /~\
R2a R3a
~15
(X)n
R1a
R2~l=~ R (CI H2)P R ,R4
3 <~(L)w--(CH2)q ,N-C-CH--N--C--A--N
It is noted that the order of carrying out the foregoing
reaction schemes is not significant and it is within the skill of one skilled
in the art to vary the order of reactions to facilitate the reaction or to
avoid unwanted reaction products.
WO 94/11012 PCI/US93/10551
2~ 3
- 76 -
The growth hormone releasing compounds of Formula I
are useful in yitro as unique tools for understanding how growth
hormone secretion is regulated at the pituitary level. This includes use
in the evaluation of many factors thought or known to influence growth
hormone secretion such as age, sex, nutritional factors, glucose, amino
acids, fatty acids, as well as fasting and non-fasting states. In addition,
the compounds of this invention can be used in the evaluation of how
other hormones modify growth hormone releasing activity. For
example, it has already been established that somatostatin inhibits
growth hormone release. Other hormones that are important and in
need of study as to their effect on growth hormone release include the
gonadal horrnones, e.g., testosterone, estradiol, and progesterone; the
adrenal hormones, e.g., cortisol and other corticoids, pinephrine and
norepinephrine; the pancreatic and gastrointestinal horrnones, e.g.,
insulin, ~lllc~on, gastrin, secretin; the vasoactive intestinal peptides,
e.g., bombesin; and the thyroid hormones, e.g., thyroxine and
triiodothyronine. The compounds of Formula I c,m also be employed to
investigate the possible negative or positive feedb~ck effects of some of
the pituitary hormones, e.g., growth hormone and endorphin peptides,
on the pituitary to modify growth hormone release. Of particular
scientific importance is the use of these compounds to elucidate the
subcellular mech~nisms me~ tin~ the release of growth hormone.
The compounds of Formula I can be ~lmini~tered to
~nim~l.s, including man, to release growth hormone in vivo. For
example, the compounds can be ~lministered to commercially important
~nim~lc such as swine, cattle, sheep and the like to accelerate and
increase their rate and extent of growth, and to increase milk produc-
tion in such ~nim~l.s. In addition, these compounds can be ~lministered
to humans in vivo as a diagnostic tool to directly determine whether the
pituitary is capable of releasing growth hormone. For example, the
compounds of Formula I can be ~(lmini~tered in vivo to children.
Serum samples taken before and after such ~lminictration can be
assayed for growth hormone. Comparison of the amounts of growth
hormone in each of these samples would be a means for directly
WO 9~/11012 2 1 ~ 7 ~ 9 3 PCI/US93/lOS51
}
- 77 -
determining the ability of the patient's pituitary to release growth
hormone.
Accordingly, the present invention includes within its scope
pharmaceutical compositions comprising, as an active ingredient, at least
one of the compounds of Formula I in association with a pharmaceutical
carrier or diluent. Optionally, the active ingredient of the ph~rm~ceu-
tical compositions can comprise a growth promoting agent in addition to
at least one of the compounds of Formula I or another composition
which exhibits a different activity, e.g., an antibiotic or other
pharmaceutically active material.
Growth promoting agents include, but are not limited to,
TRH, diethylstilbesterol, theophylline, enkephalins, E series prostaglan-
dins, compounds disclosed in U.S. Patent No. 3,239,345, e.g., zeranol,
15 and compounds disclosed in U.S. Patent No. 4,036,979, e.g., sulbenox
or peptides disclosed in U.S. Patent No. 4,411,890.
A still further use of the disclosed novel substituted
dipeptide analogs is in combination with other growth hormone
secretagogues such as GHRP-6, GHRP-1 as described in U.S. Patent
20 Nos. 4,411,890; and publications WO 89/07110 and WO 89/07111 and
B-HT920 or growth hormone releasing factor and its analogs or growth
hormone and its analogs or somatomedins including IGF-l and IGF-2.
A still further use of the disclosed novel substituted dipeptide analogs is
in combination with a2 adrenergic agonists or 133 adrenergic agonists in
2 the treatment of obesity or in combination with parathyroid hormone or
bisphosphonates, such as MK-217 (alendronate), in the treatment of
osteoporosis.
As is well known to those skilled in the art, the known and
potential uses of growth hormone are varied and multitudinous. Thus,
the ~lmini~tration of the compounds of this invention for purposes of
stim~ ting the release of endogenous growth hormone can have the
same effects or uses as growth hormone itself. These varied uses of
growth hormone may be sllmm~rized as follows: stimulating growth
hormone release in elderly humans; Prevention of catabolic side effects
of glucocorticoids, treatment of osteoporosis, stimlll~tion of the immune
WO 94/1 1012 PCI/US93/~0551
~,~4~ ~
- 78 -
system, treatment of retardation, acceleration of wound healing,
accelerating bone fracture repair, treatment of growth retardation,
treating renal failure or insufficiency resulting in growth retardation,
treatment of physiological short stature, including growth hormone
5 deficient children, treating short stature associated with chronic illness,
treatment of obesity and growth retardation associated with obesity,
treating growth retardation associated with Prader-Willi syndrome and
Turner's syndrome; Accelerating the recovery and reducing
hospit~li7~tion of burn patients; Treatment of intrauterine growth
o retardation, skeletal dysplasia, hypercortisolism and Cushings
syndrome; Induction of pulsatile growth hormone release; Replacement
of growth hormone in stressed patients; Treatment of osteochondrodys-
plasias, Noonans syndrome, schizophrenia, depression, Alzheimer's
disease, delayed wound healing, and psychosocial deprivation; treatment
5 of pulmonaly dysfunction and ventilator dependency; Attenuation of
protein catabolic response after a major operation; reducing cachexia
and protein loss due to chronic illness such as cancer or AIDS.
Treatment of hyperinsulinemia incllldin~ nesidioblastosis; Adjllvant
treatment for ovulation induction; To stimulate thymic development and
20 prevent the age-related decline of thymic function; Treatment of
immunosuppressed patients; Improvement in muscle strength, mobility,
m~intenance of skin thickness, metabolic homeostasis, renal hemeostasis
in the frail elderly; Stim~ tion of osteoblasts, bone remodelling, and
cartilage growth; Stimlll~tion of the immlme system in companion
25 ~nim~l~ and treatment of disorders of aging in companion ~nim~
Growth promotant in livestock; and stimulation of wool growth in
sheep.
The compounds of this invention can be ~lmini~tered by
oral, parenteral (e.g., intramuscular, intraperitoneal, intravenous or
3 subcutaneous injection, or implant), nasal, vaginal, rectal, sublingual, or
topical routes of ~(lmini~tration and can be formulated in dosage forms
appropriate for each route of ~c~mini~tration.
Solid dosage forms for oral ~lmini~tration include
capsules, tablets, pills, powders and granules. In such solid dosage
WO 94/1 1012 PCr/US93/105~1
~ 7 r~ ~ 3
- 79 -
forms, the active compound is admixed with at least one inert pharma-
ceutically acceptable carrier such as sucrose, lactose, or starch. Such
dosage forms can also comprise, as is normal practice, additional
substances other than inert diluents, e.g., lubricating agents such as
m~nesium stearate. In the case of capsules, tablets and pills, the dosage
forms may also comprise buffering agents. Tablets and pills can
additionally be prepared with enteric coatings.
Liquid dosage forms for oral ~minilstration include
pharmaceutically acceptable emulsions, solutions, suspensions, syrups,
the elixirs cont~inin~ inert diluents cornmonly used in the art, such as
water. Besides such inert diluents, compositions can also include
adjuvants, such as wetting agents, emulsifying and suspending agents,
and sweetening, flavoring, and perfuming agents.
Preparations according to this invention for parenteral
~mini~tration include sterile aqueous or non-aqueous solutions,
suspensions, or emulsions. Examples of non-aqueous solvents or
vehicles are propylene glycol, polyethylene glycol, vegetable oils, such
as olive oil and corn oil, gelatin, and injectable organic esters such as
ethyl oleate. Such dosage forms may also contain adjuvants such as
preserving, wetting, emulsifying, and dispersing agents. They may be
sterilized by, for example, filtration through a bacteria-retaining filter,
by incorporating sterilizing agents into the compositions, by irradiating
the compositions, or by he~ting the compositions. They can also be
manufactured in the form of sterile solid compositions which can be
dissolved in sterile water, or some other sterile injectable medium
immediately before use.
Compositions for rectal or vaginal ~dmini~stration are
preferably suppositories which may contain, in addition to the active
substance, excipients such as cocoa butter or a suppository wax.
3 Compositions for nasal or sublingual a~lministration are
also prepared with standard excipients well known in the art.
The dosage of active ingredient in the compositions of this
invention may be varied; however, it is necessary that the amount of the
active ingredient be such that a suitable dosage form is obtained. The
WO 94/1 1012 PCr/US93/10551
4~ ~3
- 80 -
selected dosage depends upon the desired therapeutic effect, on the route
of a-lmini~tration, and on the duration of the treatment. Generally,
dosage levels of between 0.0001 to 100 mg/kg. of body weight daily are
~lminictered to patients and ~nim~l~, e.g., m~mm~l~, to obtain effective
5 release of growth hormone.
The following examples are provided for the purpose of
further illustration only and are not intended to be limitations on the
disclosed invention.
o EXAMPLE 1
(R)-a-[(3-Amino-3-methyl- 1 -oxobutyl)amino] -N-l [2'-(1 H-tetrazol-5 -
yl)r l ~ l '-biphenyll -4-yllmethyllbenzene-bllt~n~mide, trifluoroacetate
Step A: 5-Phenyltetrazole
Z~nc chloride (3.3 g, 24.3 mmol, 0.5 eq) was added to 15
mL of N,N-dimethylformamide in small portions ~hile m~int~inin~ the
temperature below 60C. The sllspension of zinc chloride was cooled to
room temperature and treated with S.0 g of benzonitrile (48.5 mmol,
1.0 eq) followed by 3.2 g of sodium azide (48.5 mmol, 1.0 eq). The
heterogeneous mixture was heated at 115C with agitation for 18 hours.
The mixture was cooled to room temperature, water (30 mL) was added
and the mixture acidified by the addition of 5.1 ml_ of concentrated
hydrochloric acid. The mixture was cooled to 0C and aged for one
hour, then filtered and the filter cake washed with 15 mL of cold 0.1N
HCl then dried at 60C under vacuum to afford 6.38 g (43.7 mmol,
90%) of the product.
Step B: 5-Phenyl-2-trityltetrazole
3 To a suspension of 5.0 g (34.2 mmol) of 5-phenyltetrazole
in 55 mL of acetone was added 5.0 mL of triethyl,lmine (3.6 g, 35.6
mrnol, 1.04 eq). After 15 minutes, a solution of 10.0 g of triphenyl-
methyl chloride (35.9 mmol, 1.05 eq) in 20 mL of tetrahydrofuran was
added and the mixture stirred at room temperature for one hour.
WO 94/11012 PCI/US93/10551
21~7~
- 81 -
Water (75 mL) was slowly added and the mixture stirred for one hour
at room temperature. The product was collected by filtration, washed
with 75 mL of water and dried at 60C under vacuum to give 13.3 g
(34.2 mmol, 100%) of the product.
Step C: N-Triphenylmethyl-5-r2-(4'-methylbiphen-4-yl)ltetrazole
A solution of zinc chloride (6.3 g, 46.2 mmol, 0.6 eq) in 35
mL of tetrahydrofuran was dried over molecular sieves. 5-Phenyl-2-
trityltetrazole (30.0 g, 77.3 mmol, 1.0 eq) was dissolved in 300 mL of
dry tetrahydrofuran and the solution gently stirred while being degassed
three times by alternating vacuum and nitrogen purges. The stirred
solution was cooled to -15C and treated slowly with 50.5 mL of 1.6 M
n-butyllithium in hexane (80.0 mmol, 1.05 eq) so as to m~int~in the
temperature below -5C. The solution was m~int~ined at -5 to -15C
15 for 1.5 hours then treated with the dried zinc chloride solution and
allowed to warm to room temperature.
In a separate flask, 4-iodotoluene (20.17 g, 92.5 mmol, 1.2
eq) and bis(triphenylphosphine)nickel(II)dichloride (1.5 g, 2.3 mmol,
0.03 eq) were dissolved in 60 mL of tetrahydrofuran, then degassed and
20 left under an atmosphere of nitrogen. The mixture was cooled to 5C
and treated with 1.5 mL of 3.0 M solution of methylm~gnesium
chloride in tetrahydrofuran (4.5 mmol, 0.06 eq) so as to keep the
temperature below 10C. The solution was warmed to room tempera-
ture and added, under nitrogen purge, to the arylzinc solution. The
25 reaction mixture was stirred vigorously for 8 hours at room tempera-
ture then quenched by the slow addition of a solution of 10 mL of
glacial acetic acid (1.6 mmol, 0.02 eq) in 60 mL of tetrahydrofuran at a
rate so that the temperature was m~int~ined below 40C. The mixture
was stirred for 30 minutes and 150 mL of 80% saturated aqueous
30 sodium chloride was added; the reaction mixture was extracted for 30
minutes and the layers allowed to separate. The organic layer was
removed and washed with 150 mL of 80% saturated aqueous sodium
chloride buffered to pH >10 by the addition of arnmonium hydroxide.
The organic phase was removed and concentrated under vacuurn to
WO 9~/11012 PCIJUS93/10551
'~ - 82 -
approximately 50 mL then 250 mL of acetonitrile was added. The
mixture was again concentrated under vacuum to S0 mL and acetonitrile
added to make the final volume 150 mL. The resulting slurry was
cooled at 5C for 1 hour then filtered and washed with S0 mL of cold
5 acetonitrile followed by lS0 mL of distilled water. The filter cake was
air dried to a free flowing solid then further dried
under vacuum at 50C for 12 hours to afford 30.0 g (62.8 mmol, 81 %)
of the product. lH NMR (200 MHz, CDC13): 2.?8 (s,3H), 6.9-7.05 (m,
lOH), 7.2-7.5 (m, 12H), 7.9 (m, lH).
Step D: N-Triphenylmethyl-5-[2-(4'-bromomethylbiphen4-
yl)ltetrazole
A solution of 3.15 g (6.6 mmol) of N-triphenylmethyl-S-
[2-(4'-methylbiphen-4-yl)]tetrazole in 25 mL of mlethylene chloride was
treated with 1.29 g (7.25 mmol, 1.1 eq) of N-bronnosuccinimide, 80 mg
(05 mmol, 0.07 eq) of ArsN~ 200 mg of sodium ~cetate and 200 mg of
acetic acid. The mixture was heated at reflux for 16 hours then cooled
arld washed with saturated aqueous sodium bicarbonate. The organic
layer was removed, dried over sodium sulfate, fil~ered and concentrated
to a minimllm volume by atmospheric distillation. Methyl t-butyl ether
was added and distillation continued until almost all the methylene
chloride was removed the the total volume reduced to approximately 12
mL and 12 mL of hexanes was then added. The mixture was kept at
room temperature for 2 hours and the product isolated by filtration,
washed with hexanes then dried under vacuum at 50C to give 2.81 g
(5.04 mmol, 76%) of the product. lH NMR (200 MHz, CDC13): 4.38
(s, 2H), 6.9-8.0 (m, 23H). NMR indicates presence of approximately
1 % of the starting material and 7% of the dibromo derivative.
Step E: N-Triphenylmethyl-5-[2-(4'-azidomethylbiphen-4-
yl)ltetrazole
To 5.57 g (10 mmol) of N-triphenylrnethyl-5-[2-(4'-
bromomethylbiphen-4-yl)]tetrazole in 20 mL of dimethyl sulfoxide was
added 614 mg (12.5 mmol, 1.25 eq) of pulverized lithium azide. The
WO 94/1 1012 PCI/US93/~05~1
2I~7~3
- 83 -
reaction was stirred at room temperature for 4 hours, during which
time a thick precipitate formed. The precipitated solids were collected
by filtration and washed with methanol, water, and then methanol again,
and dried under vacuum for 16 hours to yield 4.06 g (78%) of the
product as a white solid. lH NMR (200 MHz, CDC13): 3.46 (s, 2H),
6.82-7.55 (m, 22H), 7.95 (m, lH).
Step F: N-Triphenylmethyl-5-[2-(4'-aminomethylbiphen-4-
yl)ltetrazole
To a solution of 4.06 g (7.8 mmol) of N-triphenylmethyl-
5-[2-(4'azidomethylbiphen-4-yl)]-tetrazole in 15 mL of tetrahydrofuran
was added 2.05 g (7.8 mmol, l eq) of triphenylphosphine in small
portions. The mixture was stirred at room temperature for 2 hours, at
which time 0.2 mL of water was added and the reaction mixture stirred
for 16 hours. The reaction mixture was concentrated to dryness under
vacuum and the crude product chromatographed on a silica flash
column, eluting with chloroform, to give 1.5 g (3.03 mmol, 39%) of
the product. 1H NMR (200 MHz, CDC13): 2.21 (br s, 2H), 3.75 (s, 2H),
6.80-7.94 (m, 22H), 7.94 (m, lH).
Step G: (R)-oc-[t-Butoxycarbonylamino]-N-[[2'-(~-triphenyl-
methyl-tetrazol-5-yl)[ l,1 '-biphenyl]-4-yl]methyl]benzene-
butanamide
To a solution of 30.5 mg (0.11 mmol) of N-BOC-D-
25 homophenyl~l~nine in 1 mL of methylene chloride at room temperature
under a nitrogen atmosphere was added 54 mg (0.11 mmol, l eq) of N-
triphenylmethyl-5-[2-(4'aminomethylbiphen-4-yl)]tetrazole (Step F), 25
mg (0.13 mmol, 1.2 eq) of 1-(3-dimethylaminopropyl)-3-ethylcarbodii-
mide hydrochloride and 13 mg (0.13 mmol, 1.2 eq) of triethyl~mine.
The reaction was stirred at room temperature for 16 hours, then
transferred to a separatory funnel and washed with 2 mL of 5% aqueous
citric acid and 2 mL of saturated aqueous sodium bicarbonate. The
organic layer was removed, dried over magnesium sulfate, filtered and
evaporated to dryness under vacuum. The residue was chromato-
WO 94/11012 PCI/US93/10551
- 84 -
graphed on a silica flash column, eluting with hexane/ethyl acetate (5:1),
to give 29 mg (35%) of the product. lH NMR (200 MHz, CDC13):
1.43 (s, 9H), 2.05 (m, 2H), 2.67 (t, 8Hz, 2H), 4.0~ (m, lH), 4.32 (m,
lH), 4.99 (m, lH), 6.19 (m, lH), 6.78-7.15 (m, 27H), 7.95 (m, lH).
Step H: (R)-a- [t-Butoxycarbonylamino] -N- [ [2'-(1 H-tetrazol-5 -
yl)r 1.1 '-biphenyll-4-yllmethyll-benzene-b-1t~n~mide
A solution of 29 mg (0.038 mmol) o~ the intermediate
obtained in Step G in 1 mL of methanol was hydrogenated at room
o temperature and one atmosphere over 4 mg of 20% palladium
hydroxide on carbon for two hours. The reaction mixture was then
filtered through Celite to remove the catalyst, the solvent removed
under vacuum and the residue flash chromatographed on silica to yield
19 mg (95%) of the title compound. lH NMR (200 MHz, CD30D):
1.43 (s~ 9H), l.9S (m, 2H), 2.64 (m, 2H), 4.00 (m, lH), 4.34 (t, 5Hz,
2H), 7.00-7.30 (m, 8H), 7.50 (m, SH). FAB-MS: calculated for
C2gH32N603 512; found 513 (M+l, 24%).
Step I; (R)-a-Amino-N-[[2'-(1 H-tetrazol-5-yl)[ 1,1 '-biphenyl]-
4yllmethyll-benzenebut~n~mide. hydrochloride
A solution of 19 mg (0.037 mmol) of the intermediate
obtained in Step H in 1 mL of methanol was treated with one drop of
concentrated hydrochloric acid. The mixture was stirred at room
temperature for 16 hours then evaporated to dryness under vacuum.
5 The crude product was purified on reverse phase HPLC on C18, eluting
with methanol/0.1% aqueous trifluoroacetic acid ~linear gradient: 60%
me~anol to 80% methanol over 10 minlltes], to yield 14 mg (84%~ of
the product. lH NMR (200 MHz, CD30D): 2.11 (m, 2H), 2.64 (m,
2H), 3.89 (t, 6Hz, lH), 4.42 (s, 2H), 7.05-7.32 (m, 8H), 7.58 (m, SH).
30 FAB-MS: calculated for C24H24N6O 412; found 413 (M+1,100%).
Step J: 2 2-Dimethylbutanedioic acid~ 4-methyl ester
2,2-Dimethylsuccinic acid (20 g, 137 mmol) dissolved in
200 mL of absolute methanol at 0C was treated clropwise with 2 mL of
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concentrated sulfuric acid. After the addition was complete, the
mixture was allowed to warrn to room temperature and stir for 16
hours. The mixture was concentrated under vacuum to 50 mL and
slowly treated with 200 mL of saturated aqueous sodium bicarbonate.
5 The mixture was washed with hexane (3x) and the aqueous layer
removed and cooled in an ice bath. The mixture was acidified to pH 2
by slow addition of 6N HCI then extracted with ether (8x). The
combined extracts were washed with brine, dried over magnesium
sulfate, filtered and solvents removed under vacuum. The residue was
dried at room temperature under vacuum to afford 14.7 g (91.8 mmol,
67%) of the product as a viscous oil that slowly solidified upon
standing. lH NMR (200 MHz, CDC13): 1.29 (s, 6H), 2.60 (s, 2H), 3.65
(s, 3H).
Step K: 3-Benzyloxycarbonylamino-3-methylbutanoic acid, methyl
ester
To 14.7 g (91.8 mmol) of 2,2-dimethylbutanedioic acid-4-
methyl ester in lS0 mL of benzene was added 13 mL of triethylamine
(9.4 g, 93 mmol) followed by 21.8 mL of diphenylphosphoryl azide
20 (27.8 g, 101 mmol). The mixture was heated under nitrogen at reflux
for 45 minutes then 19 mL (19.9 g, 184 mmol) of benzyl alcohol was
added and refluxing continued for 16 hours. The mixture was cooled,
filtered and the filtrate concentrated to a minimum volume under
vacuum. The residue was redissolved in 250 mL of ethyl acetate,
25 washed with water, saturated aqueous sodium bicarbonate (2x) and
brine. The organic layer was removed, dried over m~nesium sulfate,
filtered and the filtrate concentrated to a minimllm volume under
vacuum. The crude product was purified by medium pressure liquid
chromatography on silica, eluting with hexane/ethyl acetate (4:1), to
afford 18.27 g (68.9 mmol, 75%) of the product. lH NMR (200 MHz,
CDCl3): 1.40 (s, 6H), 2.69 (s, 2H), 3.63 (s, 3H), S.OS (s, 2H), 5.22 (br
s, lH), 7.32 (s, 5H).
Step L: 3-Benzyloxvcarbonylamino-3-methylbutanoic acid
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A solution of 18.27 g (68.9 mmol) of 3-benzyloxycar-
bonylamino-3-methylbutanoic acid methyl ester in 20 mL of methanol
at room temperature was treated dropwise with 51 mL of 2N NaOH
(102 mmol). The mixture was stirred at room temperature for 16
5 hours then transferred to a separatory funnel and washed with hexane
(3x). The aqueous layer was removed, cooled to 0C and slowly
acidified to pH 2 (paper) by dropwise addition of 6N HCl. This mixture
was extracted with ether (6x); combined extracts were washed with lN
HCl and brine, then dried over magnesium sulfate, filtered and solvent
removed under vacuum to afford 17.26 g (68.7 mmol, 99%) of the
product. lH NMR (200 MHz, CDCl3): 1.42 (s, 6H), 2.77 (s, 2H), 5.06
(s, 2H), 5.2 (br s, lH), 7.3 (s, SH).
Step M: 3-Benzyloxycarbonylamino-3-methylbutanoic acid, N-
hydroxysuccinimide ester
A solution of 2.93 g (11.7 mrnol, 0.~5 eq) of 3-benzyl-
oxycarbonylamino-3-methylbutanoic acid in S mL of methylene
chloride at room temperatllre was treated with 1.61 g (14.0 mmol) of
N-hydroxysuccinimide followed by 2.67 g (14.0 Enmol, l eq) of 1(3-
20 dirnethylaminopropyl)-3-ethylcarbodiimide hydrochloride. The
reaction mixture was stirred for 16 hours then tr~msferred to a
separatory furmel, washed with water and dilute aqueous sodium
bicarbonate. The organic layer was removed, dried over magnesium
sulfate, filtered, and solvents removed under vacuum. The crude
25 product was chromatographed on a silica flash column, eluting with
hexane/ethyl acetate (1:1), to give 3.9 g (qll~ntit~tive) of the product.
1H NMR (200 MHz, CDCl3): 1.51 (s, 6H), 2.80 (s, 4H), 3.12 (s, 2H),
5.13 (s, 2H), 7.37 (s, SH). FAB-MS: calculated f~r C17H20N2o6 348;
found 349 (M+l, 40%).
Step N: (R)-a-[(3-Benzyloxycarbonylamino-3-methyl-1-oxo-
butyl)amino]-N-[[2'-(1 H-tetrazol-S-yl)-[ l, l '-biphenyl]-
4yllmethyllbenzenebutanamide
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A solution of 12 mg (0.023 mmol) of the intermediate
obtained in Step I in 0.5 mL of methylene chloride at room temperature
was treated with 9.2 mg (0.027 mmol, 1.2 eq) of 3-benzyloxy-
carbonylamino-3-methylbutanoic acid, N-hydroxysuccinimide ester
(Step M) and 3.5 mg (0.027 mmol, 1.2 eq) of diisopropylethyl~mine.
The reaction mixture was stirred at room temperature for 48 hours.
Solvents were emoved under vacuum and the crude residue was purified
on HPLC to give 14 mg of product. lH NMR (200 MHz, CD30D):
1.29 (s, 3H), 1.36 (s, 3H), 1.90 (m, 3H), 2.45 (m, 3H), 4.31 (m, lH),
4.72 (d, 20Hz, lH), 5.02 (d, 20Hz, lH), 7.00-7.62 (m, 18H). FAB-MS:
calculated for C37H39N704 645; found 646 (M+l, 100%).
Step O: (R)-oc-[(3-Amino-3-methyl- 1 -oxobutyl)amino] -N-[[2'-(1 H-
tetrazol-5-yl)[1,1 '-biphenyl] -4-yl]methyl]benzenebutana-
mide~ tri-fluoroacetate
A solution of 14 mg (0.21 mmol) of the intermediate
obtained in Step N in 1 mL of methanol was hydrogenated at room
temperature and one atmosphere over l mg of 20% palladium
hydroxide on carbon for 16 hours. The reaction mixture was filtered
20 through Celite and the filtrate concentrated under vacuum and the
residue chromatographed on reverse phase high pressure liquid
chromatography on C18, eluting with methanol/0.1% aqueous
trifluoroacetic acid (linear gradient; 65% methanol increased to 85%
over 10 minllt~s), to give 10 mg (72%) of the title compound. lH NMR
25 (200 MHz, CD30D): 1.32 (s, 3H), 1.36 (s, 3H), 1.81 (m, lH), 1.95 (m,
2H), 2.26 (m, lH), 2.52 (s, lH), 2.63 (m, lH), 4.34 (m, lH), 4.76 (d,
14Hz, lH), 4.95 (d, 14Hz, lH), 7.01-7.70 (m, 13H). FAB-MS:
calculated for C29H33N7O2 511; found 512 (M+l, 100%).
E~XAMPLE 2
(R)-a-[(3-Amino-3-methyl- 1 -oxobutyl)amino] -N-[[2'-(1 H-tetrazol-5-
yl)rl.l'-biphenvll-4-yllmethyllbenzene-propanamide. trilCluoroacetate
wo 94/11012 Pcr/US93/~0551
88-
The title compound was prepared from N-BOC-D-
phenylalanine by the methods described in Example 1. lH NMR (200
MHz, CDCl3): 1.27 (s, 3H), 1.40 (s, 3H), 2.49 (d, l5Hz, lH), 2.62
(d, 15Hz, lH), 3.10 (m, 2H), 4.44 (m, 2H), 4.80 (m, lH), 7.16 (s, 4H),
7.35 (s, 5H); 7.70 (m, 4H). FAB-MS: calculated ~or C28H3 lN702 497;
found 498 (M+1,100%).
13XAMPLE 3
(R)-oc-[(3-Amino-3-methyl-1-oxobutyl)amino]-N-[[2'-(lH-tetrazol-5-
yl)[1,1 '-biphenyl]-4-yl]methyl]- 1 H-indole-3-propanamide,
trifluoroacetate
Step A: (R)-oc-[t-Butoxycarbonylamino]-N-[12'-(~-triphenyl-
me~yl-tetrazol-5-yl)[ 1,1 '-biphenyl]- ~-yl]methyl]-(Nim-
fo~nyl)indole-3 -propanamide
Prepared from N-triphenylmethyl-5-[2-(4'-aminomethyl-
biphen-4-yl)]tetrazole and Nim-fo~nyl-Na-BOC D-tryptophan by the
procedure described in E~xarnple 1, Step G. lH NMR (200 MHz,
CDCl3): 1.40 (s, 9H), 3.18 (br s, 2H), 4.24 (br s, 2H), 4.48 (m, lH),
5.24 (s, lH), 6.28 (s, lH), 6.70-7.70 (m, 27H), 7.90 (m, lH), 8.82 (br s,
lH).
Step B: (R)-a-[t-Butoxycarbonylamino]-N-[[2'-(lH-tetrazol-5-
yl)[1,1 '-biphenyl] -4-yl]methyl]-(Nim-formyl)indole-3-
propanamide
Prepared from the intermediate obtained in Step A by the
procedure described in Example 1, Step H. lH NMR (200 MHz,
CD30D): 1.38 (s, 9H), 3.14 (m, 3H), 6.90-7.70 (m, 13H), 9.02 (br s,
30 lH). FAB-MS: calculated for C30H31N704 553; found 554
(M+ 1,40%).
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Step C: (R)-o~-Amino-N-[[2'-(1 H-tetrazol-5-yl)[ l,1 '-biphenyll-4-
yl]methyl] -(Nim-formyl)indole-3 -propanamide,
trifluoroacetate
A solution of 80 mg (0.14 mmol) of the intermediate
5 obtained in Step B in 4 mL methylene chloride was treated with l mL of
anisole followed by 4 mL of trifluoroacetic acid. The reaction mixture
was stirred at room temperature for 1 hour, then all volatiles were
removed under vacuum. The residue was purified by reverse phase
high pressure liquid chromatography on C18, eluting with
methanol/0.1% aqueous trifluoroacetic acid (45:55), to give 64 mg
(77%) of the product. lH NMR (200 MHz, CD30D): 3.26 (m, 2H),
4.11 (t, 6Hz, lH), 4.28 (m, 2H), 6.99 (s, 4H), 7.28-7.70 (m, 9H), 8.75
(br s, lH). FAB-MS: calculated for C26H23N7O2 465; found 466
(M+1, 22%).
Step D: 4.4-Dimethylazetidin-2-one
A 3-neck 3L round bottom flask equipped with a magnetic
stirrer, thermometer, cold finger condenser and nitrogen bubbler was
charged with lL of ether. The flask was cooled to -65C and into it was
20 condensed 500-600 mL of isobutylene. The cold finger condenser was
replaced with a dropping funnel and 200 mL (325 g, 2.30 mol) of
chlorosulfonyl isocyanate was added dropwise over 1.5 hours. The
mixture was m~int~ined at -65C for 1.5 hours then the dry ice/acetone
cooling bath replaced with methanol/ice and the internal temperature
25 slowly increased to -5C at which time the reaction initi~ted and the
internal temperature rose to 15C with evolution of gas. The internal
temperature remained at 15C for several minutes then dropped back
down to -5C and the mixture stirred at -5C for 1 hour. The
methanol/ice bath was removed and the reaction mixture warmed to
30 room temperature and stirred overnight.
The reaction mixture was transferred to a 3-neck 12L
round bottom flask fitted with a mechanical stirrer and diluted with 2L
of ether. The well-stirred reaction mixture was treated with 2L of
saturated aqueous sodium sulfite. After 1 hour, an additional lL of
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saturated aqueous sodium sulfite was added followed by sufficient
sodium bicarbonate to adjust the pH to approximately 7. The mixture
was stirred another 30 minutes then the layers allowed to separate. The
ether layer was removed and the aqueous layer reextracted with 2 x lL
of ether. The combined ether extracts were washed once with 500 mL
of saturated aqueous sodium bicarbonate and once with 500 mL of
saturated aqueous sodium chloride. The ether layer was removed, dried
over m~nesium sulfate, filtered and concentrated under vacuum to give
33 g of a pale yellow oil. The aqueous layer was made basic by the
o addition of solid sodium bicarbonate and extracted with 3 x lL of ether.
The combined ether extracts were washed and dried as described above,
then combined with the original 33 g of pale yellow oil and concen-
trated under vacuum to give 67.7 g of product. ~urther extraction of
the aqueous layer with 4 x lL of methylene chlor;de and washing and
drying as before gave an additional 74.1 g of product. Still further
extraction of the aqueous layer with 4 x lL of methylene chloride gave
an additional 21.9 g of product. The combined product (163.7 g, 1.65
mol, 72%) was used in Step E without purification. lH NMR (200
MHz, CDCl3): 1.45 (s, 6H), 2.75 (d, 3Hz, 2H), 5.9 (br s, lH).
Step E: N-(t-Butoxycarbonyl)-4.4-dimethvla7etidin-2-one
A 5L, 3-neck round bottom flask equipped with a magnetic
stirrer, thermometer, nitrogen bubbler and addition funnel was charged
with 88.2 g (0.89 mol) of 4,4-dimethylazetidin-2--one, 800 mL of
methylene chloride, 150 mL of triethyl~mine (1.08 mol) and 10.9 g
(0.089 mol) of 4-dimethylaminopyridine. To the stirred solution at
room temperature was added dropwise over 15 minlltes a solution of
235 g (1.077 mol) of di-t-butyl-dicarbonate in 300 mL of methylene
chloride. The reaction mixture was stirred at room temperature
overnight then diluted with lL of methylene chlo ride and washed with
500 mL of saturated aqueous ammonium chloride, 500 mL of water,
and 500 mL of saturated aqueous sodium chloride. The organic layer
was separated, dried over magnesium sulfate, filtered and concentrated
~VO 94J11012 21 ~ 7~ D 3 PCT/US93/10~1
- 91 -
under vacuum to afford 180.3 g of crude product as an orange solid.
The material was used directly in Step F without purification.
1H NMR (200 MHz, CDC13): 1.50 (s, 9H), 1.54 (s, 6H), 2.77 (s, 2H).
5 Step F: 3-t-Butoxycarbonylamino-3-methylbutanoic acid
A 3L, 3-neck round bottom flask equipped with a magnetic
stirrer, thermometer, nitrogen bubbler and addition funnel was charged
with 180.3 g (0.89 mol) of N-(t-butoxycarbonyl)-4,4-dimethylazetidin-
2-one dissolved in lL of tetrahydrofuran. The solution was cooled to 0-
o 5C and treated dropwise with 890 mL of l.OM aqueous lithiumhydroxide over 30 minutes. The reaction mixture was stirred at 0-5C
for 2 hours then diluted with lL of ether and lL of water. The layers
were allowed to separate and the aqueous layer was reextracted with an
additional lL of ether. The aqueous layer was acidified by the addition
5 of lL of saturated aqueous sodium bisulfate, then extracted with 1 x lL
and 2 x 500 mL of ether. The combined organic layer and ether
extracts were washed with 500 mL of saturated aqueous sodium
chloride, dried over magnesium sulfate and concentrated under vacuum
to give 173 g of a yellow oil that solidified upon standing. The material
20 was slurried with warm hexane then filtered and dried under high
vacuum to afford 168.5 g (0.775 mol, 87%) of product as a white solid.
lH NMR (200 MHz, CDC13): 1.39 (s, 6H), 1.44 (s, 9H), 2.72 (s, 2H).
FAB-MS: calculated for C1oH1gNO4 217; found 218 (M+H, 54%).
25 Step G: (R)-oc-[(3-t-Butoxycarbonylamino-3-methyl-1-oxo-
butyl)amino]-N-[[2'-(lH-tetrazol-5-yl)-[1,1 '-biphenyl]-4-
yllmethvll -(Nim -formyl)-indole-3-propanamide
A solution of 64 mg (0.11 mmol) of the intermediate
obtained in Step C in 1 mL of methylene chloride at 0C was treated
30 with 25 mg (0.12 mmol, 1.1 eq) of dicyclohexylcarbodiimide and the
resulting solution stirred at 0C for 30 minutes. A solution of 53 mg
(0.24 mmol, 2 eq) of 3-t-butoxycarbonylamino-3-methylbutanoic acid
and 12 mg (0.12 mmol, 0.017 mL, 1.1 eq) of triethylamine in 1 mL of
methylene chloride was added and the mixture stirred for 5 hours al
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room temperature. The reaction mixture was evaporated to dryness
under vacuum and the residue was dissolved in 1 mL of anisole and
treated with 4 mL of trifluoroacetic acid. The mixture was stirred at
room temperature for 30 minutes the concentrated under vacuum. The
5 residue was taken up in methanol and the solids removed by filtration.
The filtrate was concentrated under vacuum; the residue was purified by
reverse phase high pressure liquid chromatography on C18 to yield 31
mg (42%) of the product. lH NMR (200 MHz, CD30D): 1.14 (s, 3H),
1.25 (s, 3H), 2.37 (d, 15Hz, lH), 2,48 (d, 15Hz, l ~I), 3.11 (m, 2H), 4.26
(m, 2H), 4.72 (m, lH), 6.92-7.70 (m, 13H), 8.55 ~br s, lH).
FAB-MS: calculated for C31H32NgO3 564; found 565 (M+1, 80%).
Step H: (R)-a-[(3-Amino-3-methyl- 1 -oxobutyl)amino]-N-[[2'-(1 H-
tetrazol-5 -yl) [1,1 '-biphenyl] -4-yl]methyl] - 1 H-indole-3 -
propanamide. trifluoroacetate
A solution of 31 mg (0.046 mmol) of the intelmediate
obtained in Step G in 1 mL of methanol was treated with 0.2 mL of
concentrated hydrochloric acid and the resulting mixture heated at 65C
for 1.5 hours. All volatiles were removed under ~acuum and the
20 residue purified by reverse phase high pressure liquid chromatography
on C18, eluting with methanol/0.1% aqueous trifluoroacetic acid
(50:50), to give 20 mg (67%) of the title compound. lH NMR (200
MHz, CD30D): 1.15 (s, 3H), 1.28 (s, 3H), 2.38 (d, 16Hz, lH), 2.50 (d,
16Hz, lH), 3.19 (m, 2H), 4.20 (d, 14Hz, lH), 4.32 (d, 14Hz, lH), 4.73
5 (t, 7Hz, lH), 6.90-7.70 (m, 13H). FAB-MS: calculated for
C30H32N8o2 536; found 536 (70%).
EXAMPLE 4
(R)-2-[(3-Amino-3-methyl- 1 -oxobutyl)amino]-N-phenyl-N-[ [2'-(1 H-
tetrazol-5-yl)r l . l '-biphenyll-4-yllmethyll-butanamide~ trifluoroacetate
Step A: (R)-2-(t-Butoxycarbonylamino)-N-phenyl-butanamide
WO 94/11012 PCI/US93/10551
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~ 7 ~ ~ ~
- 93 -
Prepared from N-BOC-D-2-arninobutanoic acid and aniline
by the method described in Example 1, Step G. lH NMR (200 MHz,
CDC13): 1.00 (t, 6Hz, 3H), 1.45 (s, 9H), 1.82 (m, 2H), 4.30 (m, lH),
5.59 (d, 7Hz, lH), 7.00-7.62 (m, 5H), 8.90 (br s, lH). FAB-MS:
calculated for C15H22N2O3 278; found 279 (M+1, 40%).
Step B: (R)-2-(t-Butoxycarbonylamino)-N-phenyl-N-[[2'-(~-
triphenylmethyl-tetrazol-5-yl)-[1,1'-biphenyl] -4-yl]methyl] -
butanamide
A solution of 70 mg (0.25 mmol) of (R)-2-(t-butoxy-
carbonylamino)-N-phenyl-but~n~mide in 0.5 mL of dry dimethyl-
formamide was treated with 10 mg (0.25 mmol, l eq) of 60% sodium
hydride oil dispersion. The mixture was stirred at room temperature
for 20 minutes, then treated with a solution of 140 mg (0.25 mmol, l
5 eq) of N-triphenylmethyl-5-[2-(4'-bromomethylbiphen-4-yl)]tetrazole
(Example 1, Step D) in 0.5 mL of dry dimethylformamide. The
mixture was stirred at room temperature for 2 hours, then quenched by
the addition of 2 mL of water. The mixture was extracted several times
with ethyl acetate; the combined extracts were washed with 5% aqueous
citric acid, saturated aqueous sodium bicarbonate then dried over
magnesium sulfate, filtered and the filtrate dried under vacuum. The
crude material was chromatographed on a flash silica column, eluting
with hexane/ethyl acetate (1:1), to give 91 mg (48%) of the product. lH
NMR (200 MHz, CDC13): 0.74 (t, 6Hz, 3H), 1.32 (s, 9H), 1.58 (m,
25 2H), 4.20 (m, lH), 4.79 (s, 2H), 6.82-7.60 (m, 27H), 7.86 (m, lH).
Step C: (R)-2-(t-Butoxycarbonylamino)-N-phenyl-N-[[2'-(lH-
tetrazol-5-yl)-~1.1'-biphenyll -4-yllmethyllbutanamide
Prepared from the intermediate obtained in Step B by the
3 procedure described in Example 1, Step H. 1 H NMR (200 MHz,
CDC13): 0.74 (t, 6Hz, 3H), 1.36 (s, 9H), 1.55 (m, 2H), 4.13 (m, lH),
4.70 (d, 14Hz, lH), 5.02 (d, 14Hz, lH), 5.22 (m, lH), 7.00-7.62 (m,
12H), 8.01 (m, lH). FAB-MS: calculated for C29H32N6O3 512; found
513 (M+l, 100%).
~VO 9~/11012 PCT/US93/lOS;i
Q3 94
Step D: (R)-2-Amino-N-phenyl-N-[[2'-(1H-tetrazol-5-yl)[l,l'-
biphenyll-4-yllmethyllbut~n~mide. hydrochloride
Prepared from the intermediate obtained in Step C by the
method described in Example 1, Step I. 1H NMR (200 MHz, CD30D):
0.83 (t, 7Hz, 3H), 1.68 (m, 2H), 3.80 (m, lH), 4.81 (d, 14Hz, lH), 5.01
(d, 14Hz, lH), 7.00-7.70 (m, 13H). FAB-MS: calculated for
C24H24N60 412; found 413 (M+1, 100%).
Step E: 3-t-Butoxycarbonylamino-3-methylbutanoic acid, N-
hydroxysuccinimide ester
Prepared from 3-t-butoxycarbonylamino-3-methylbutanoic
acid (Example 3, Step F) and N-hydroxysuccinimde by the procedure
describe in Example 1, Step M. lH NMR (200 MHz, CDCl3): 1.41 (s,
9H), 1.43 (s, 6H), 2.82 (s, 4H), 3.07 (s, 2H), 4.72 (br s, lH).
Step F: (R)-2-[(3-t-Butoxycarbonylamino-3-methyl-1-oxobutyl)-
amino]-N-phenyl-N-[ [2'-(1 H-tetra-zol-5 -yl) [1,1 '-biphenyl] -
4yllmethyll -but~n~mi de
Prepared as in Example 1, Step N from (R)-2-amino-N-
phenyl-N-[[2'-(lH-tetrazol-5-yl)[l,l'-biphenyl]-4-yl]methyl]b~lt~n~mide,
hydrochloride and 3-t-butoxycarbonylamino-3-methylbutanoic acid, N-
hydroxysuccinimide ester. lH NMR (200 MHz, CD30D): 0.74 (t, 7Hz,
3H), 1.32 (s, 6H), 1.40 (s, 9H), 1.58 (m, 2H), 2.4S (d, 13Hz, lH), 2.58
(d, 13Hz, lH), 4.29 (m, lH), 4.78 (d, 14Hz, lH), 4.9~ (d, 14Hz, lH),
6.97-7.79 (m, 13H). FAB-MS: calculated for C34H41N704 611; found
612 (M+l, 100%).
Step G: (R)-2-[(3-Amino-3-methyl-1-oxobutyl)amino]-N-phenyl-N-
[[2'-(lH-tetrazol-5-yl)[ l,1 '-biphenyl] 4-yl]methyl]-
butanamide. -trifluoroacetate
The title compound was prepared from the intermediate
obtained in Step F by the procedure described in Example 3? Step H.
1H NMR (200 MHz, CD30D): 0.72 (t, 7Hz, 3H), 1.32 (s, 3H), 1.40 (s,
WO 94/1 1012 PC~/US93/~05~1
77~
3H), 1.58 (m, 2H), 2.51 (m, 2H), 4.28 (m, lH), 4.73 (d, 14Hz, lH),
4.99 (d, 14Hz, 1 H), 6.95-7.83 (m, 1 3H). FAB-MS: calculated for
C29H33N7O2 51 1; found 512 (M+1, 100%).
E~AMPLE 5
2-[(3-Amino-3-methyl- 1 -oxobutyl)amino]-N-[[2'-(1 H-tetrazol-5-
yl)r1.1'-biphenyll-4-yllmethyllacetamide. trifluoroacetate
Step A: 2-t-Butoxycarbonylamino-N-[[2'-(~-triphenylmethyl-
tetrazol-5-yl)~ 1~1 '-biphenyll-4-yll-methyllacetamide
Prepared from N-triphenylmethyl-5-[2-(4'-aminomethyl-
biphen-4-yl)]tetrazole (Example 1, Step F) and N-BOC-glycine by the
procedure described in Example 1, Step G. lH NMR (200 MHz,
CDC13): 1.43 (s, 9H), 3.72 (d, SHz, 2H), 4.32 (d, 6Hz, 2H), 5.06 (m,
lH), 6.30 (m, lH), 6.82-7.68 (m, 22H), 7.95 (m, lH).
Step B: 2-t-Butoxycarbonylamino-N-[[2'-(1H-tetrazol-5-yl)[1,1'-
biphenyll -4-yllmethyll acetamide
Prepared from the intermediate obtained in Step A by the
procedure described in Example 1, Step H. 1H NMR (200 MHz,
CD30D): 1.42 (s, 9H), 3.69 (s, 2H), 4.38 (d, 6Hz, 2H), 6.90-7.28 (m,
4H), 7.42-7.69 (m, 4H). FAB-MS: calculated for C21H24N603, 408;
found 409 (M+1, 20%).
Step C: 2-Amino-N-[[2'-(1 H-tetrazol-5-yl)[ 1,1 '-bi-phenyl]-4-
yllmethyllacetamide~ hydrochloride
Prepared from the intermediate obtained in Step B by the
procedure described in Example 1, Step I. lH NMR (200 MHz,
30 CD30D): 3.84 (s, 2H), 4.35 (s, 2H), 7.10-7.83 (m, 8H). FAB-MS:
calculated for C1 6H1 6N60 308; found 309 (M+1, 100%).
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Step D: 2-[(3-Benzyloxycarbonylamino-3-melthyl-1-oxo-butyl)-
amino] -N-[[2'-( l H-tetrazol-5-yl)[1,1 '-biphenyl]-4yl] -
methyll acetamide
Prepared from the intermediate obtained in Step C and 3-
5 benzyloxycarbonylamino-3-methylbutanoic acid, N-hydroxysuccinimide
ester by the procedure described in Example 1, Step N. lH NMR (200
MHz, CD3OD): 1.37 (s, 6H), 2.60 (s, 2H), 3.79 (s, 2H), 4.33 (s, 2H),
5.00 (s, 2H), 6.95-7.65 (m, 13H). FAB-MS: calculated for
C26H33N7O4 507; found 508 (M+1, 20%).
Step E: 2-[(3-Amino-3-methyl-1-oxobutyl)amino]-N-[[2'-(lH-
tetrazol-5-yl)[1,1 '-biphenyl]-4-yl]methyl]acetamide,
trifluoroacetate
Prepared from the intermediate obtained in Step D by the
procedure described in Example 1, Step O. 1H N~R (200 MHz,
CD30D): 1.38 (s, 6H), 2.52 (s, 2H), 3.89 (s, 2H), 4.38 (s, 2H), 7.00-
7.70 (m, 8H). FAB-MS: calculated for C21H25N7O2 407; found 408
(M+ 1,100%).
EXAMPLE 6
(R)-o~-[(2-Arnino-2-methyl- 1 -oxopropyl)amino]-N-[[2'-[[(methyl-
amino)carbonyl]amino] [1,1 '-biphenyl] -4-yl]-methyl] - 1 H-indole-3-
propanamide~ trifluoroacetate
Step A: N-(t-Butoxycarbonyl)-D-tryptophan benzyl ester
Finely divided t-butoxycarbonyl-D-tryptophan (3 g, 10
mmol) was suspended in methylene chloride and benzyl alcohol (1.08
mL, 10 mmol) and 4-dimethylaminopyridine (0.1" g, 1 mmol) were
3 added and stirred at room temperature. Solid 1-(3-dimethylamino-
propyl)-3-ethylcarbodiimide hydrochloride (1.92 g, 10 mmol) was then
added in three roughly equal portions over 5 minutes. The reaction
mixture was stirred for 3 hours at room temperanlre during which time
the reaction mixture became a homogeneous solut;on. The reaction
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mixture was poured into water (100 mL) and extracted with methylene
chloride (2 x 50 mL). The combined methylene chloride layers were
washed with 5% aqueous citric acid solution (100 mL) and 5% aqueous
sodium bicarbonate solution (100 mL). The resulting methylene
5 chloride layer was dried over magnesium sulfate, filtered and
evaporated under vaccum to give an off-white solid. This solid material
was chromatographed on silica gel using ethyl acetate~exanes (2:3 v/v)
as eluant. This afforded 3.56 g (91%) of the desired benzyl ester as a
white amorphous powder.
lH NMR (400 MHz, CDCl3): 1.42 (s, 9H), 3.27 (d, 2H), 4.69 (m, lH),
5.17 (ABq, 2H), 6.78 (br s, lH), 7.15-7.42 (m, 8H), 7.53 (d, lH), 7.97
(br s, lH).
Step B: D-Tryptophan benzyl ester
N-(t-Butoxycarbonyl)-D-tryptophan benzyl ester (3.5 g,
8.87 mmol) was dissolved in methylene chloride (10 mL) and stirred at
room temperature and trifluoroacetic acid (20 mL) was added dropwise
to the ester. The reaction mixture was stirred at room temperature for
one hour during which time the reaction darkened. The reaction
20 mixture was directly evaporated under vacuum to give a white solid.
This solid was dissolved in chloroform (100 mL) and washed with
saturated aqueous sodium bicarbonate. The aqueous layer was extracted
with chloroform (2 x 25 mL) and the combined chloroform layers were
dried over potassium carbonate. Filtration and concentration of the
25 chloroform solution under vacuum gave a pale yellow oil (3.14 g, 82%)
which was mainly ~e desired product.
lH NMR (400 MHz, CDC13): 1.58 (s, 9H), 3.09 (dd, lH), 3.27 (dd,
lH), 3.88 (m, lH), 5.10 (s, 2H), 6.93 (br s, lH), 7.15-7.39 (m, 8H),
7.59 (d, lH), 8.03 (br s, lH).
Step C: (R)-a-[(2-t-Butoxycarbonylamino-2-methyl-1-oxopropyl)-
aminol-lH-indole-3-propanoic acid. benzyl ester
Crude D-tryptophan benzyl ester (1.0 g, 3.40 mmol), 1-
hydroxybenztriazole hydrate (0.63 g, 4.1 mmol) and t-butoxycarbonyl-
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o~-methylalanine (0.84 g, 4.11 mmol) were stirredi together at room
temperature in chloroform (20 mL). 1-(3-Dimethylaminopropyl)-3-
ethylcarbodiimide hydrochloride (980 mg, 5.11 mmol) was added to
this mixture in a single portion. The reaction mixture was stirred at
5 room temperature for 4 hours and worked up by pouring into water (50
mL). The chloroform layer was separated and washed with 5% aqueous
citric acid solution (25 mL) and 5% aqueous sodium bicarbonate
solution (25 mL). The chloroform layer was dried over magnesium
sulfate, filtered and evaporated under vacuum to 2fford a thick oily
foam. Chromatography on silica gel using ethyl acetate/hexanes (2:3
v/v) afforded a yellow foam (0.822 g 50%). lH ~R (400 MHz,
CDCl3): 1.30 (s, 9H), 1.39 (s, 6H), 3.29 (m, 2H), 4.88 (m, lH), 5.03
(s, 2H), 6.88 (br s, lH), 7.05-7.35 (m, 8H), 8.53 (d, lH), 7.98 (br s,
lH).
Step D: (R)-oc-[(2-t-Butoxycarbonylamino-2-methyl-1-oxopropyl)-
aminol - 1 H-indole-3 -propanoic acid
The benzyl ester (0.82 g, 1.71 mmol) obtained in Step C
and 10% palladium on carbon (150 mg) were stirred together in ethyl
20 acetate (5 mL). The solution was degassed and a hydrogen atmosphere
introduced over the reactants using a balloon for 32 hours. The
reaction products were isolated by filtering the reaction mixture
through a Celite plug. The plug was washed with additional ethyl
ac~tate (3 x 10 mL). The combined filtrates were evaporated under
25 vacuum to afford the product (680 mg, 102%). l~I NMR (400 MHz,
CDCl3): 1.30 (s, 9H), 1.41 (s, 6H), 3.32 (dd, lH), 3.42 (m, lH), 4.87
(br s, lH), 6.82 (d, lH), 7.13-7.35 (m, 8H), 7.60 ~d, lH), 8.28 (br s,
lH).
3 Step E: 4-Methyl-2'-nitro-1 1'-biphenyl
A vigorously stirred mixture of 34 g (0.25 mol) of 4-
tolylboronic acid and 34 g (0.17 mol) of 2-bromo-1-nitrobenzene in a
mixture of 170 mL of 5N sodium hydroxide, 57 mL of water, 215 mL
of 2-propanol and 1080 mL of benzene was treated with 11.9 g of
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(tetrakis)triphenylphosphine palladium(0). The two-phase mixture was
heated at reflux for three hours. The cooled reaction mixture was
filtered through Celite and the filter cake washed with fresh benzene.
The organic layer was separated and washed with water (3x), dried over
magnesium sulfate and filtered. The filtrate was evaporated under
vacuum and the residue (46.1 g) purified by preparative high pressure
liquid chromatography on silica gel, eluting with hexane/ethyl acetate
(20:1), to give 28.05 g of the product. lH NMR (400 MHz, CDCl3):
2.38 (s, 3H), 7.20 (m, 4H), 7.43 (m, 2H), 7.59 (t, lH), 7.8 (d, lH).
EI-MS: calculated for C13H1 lNO2 213; found 213 (M+).
Step F: 4-Bromomethyl-2'-nitro-1~1'-biphenyl
Prepared from 4-methyl-2'-nitro-1,1'-biphenyl by the
procedure described in Example 1, Step D. 1H NMR (200 MHz,
CDC13): 4.53 (s, 2H), 7.2-7.7 (m, 7H), 7.85 (m, lH).
Step G: 4-Azidomethyl-2'-nitro-1~1'-biphenyl
Prepared from 4-bromomethyl-2'-nitro-1,1'-biphenyl by
the procedure described in Example 1, Step E. lH NMR (200 MHz,
CDCl3): 4.39 (s, 2H), 7.2-7.7 (m, 7H), 7.85 (d, lH).
Step H: 4-Aminomethyl-2'-nitro-1.1'-biphenyl
Prepared from 4-azidomethyl-2'-nitro-1,1'-biphenyl by the
procedure described in Example 1, Step E. lH NMR (200 MHz,
25 CDCl3): 3.90 (s, 2H), 7.2-7.7 (m, 7H), 7.83 (d, lH).
Step I: (R)-a-[(2-t-Butoxycarbonylamino-2-methyl-1-oxopropyl)-
amino]-N-[[(2'-nitro)[ 1,1 '-biphenyl]-4-yl]methyl]-1H-
indole-3 -propanamide
The acid (338 mg, 0.87 mmol) from Step D and 4-
aminomethyl-2'-nitro-1,1'-biphenyl (200 mg, 0.87 mmol) and
triethylamine (0.245 mL, 1.76 mmol) were dissolved in methylene
chloride (8 mL) and stirred at room temperature. Benzotriazolyl-1-
yloxytris(dimethylamino)phosphonium hexafluorophosphate (388 mg,
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0.87 mmol) was added in a single portion. The reaction mixture was
stirred togerther for 3.5 hours then the reaction was quenched by
adding saturated aqueous sodium chloride (10 mL) and extracted with
methylene chloride (3 x 20 mL). The combined extracts were dried
5 over magnesium sulfate, filtered and evaporated under vacuum. The
resulting thick gum was chromatographed on silica gel using ethyl
acetate/hexanes (1 :2 v/v) to give 297 mg (50%) of an orange semi-solid.
1H NMR (400 MHz, CD3OD): 1.08 (s, 9H), 1.26 (s, 3H), 1.32 (s, 3H),
3.29 (dd, lH), 3.43 (dd, lH), 4.35 (m, 2H), 4.64 (m, lH), 7.00-7.20 (m,
7H), 7.34 (d, lH), 7.42 (m, lH), 7.53 (t, lH), 7.61 (d, lH), 7.67 (t, lH),
7.83 (d, lH), 8.22 (s, lH).
Step J: (R)-a-[(2-t-Butoxycarbonylamino-2-methyl-1-oxopropyl)-
arnino]-N-[[(2'-amino)[1,1 '-biphenyl]-4-yl]methyl]-lH-
l 5 indole-3-propanamide
The amide (142 mg, 0.24 mmol) from Step I was dissolved
in ethanol (5 mL) and 10% palladium on carbon (15 mg) was added.
The ethanolic mixture was degassed and a hydrogen atmosphere
introduced and m~int~ined above the reaction mixture for 2.5 hours
20 using a balloon. The hydrogenation catalyst was removed by filtration
through a Celite pad. The pad was washed carefu~ly with methylene
chloride (4 x 5 mL). The combined filtrates were evaporated under
vacuum to give a powdery white foam (124 mg, 92%). lH NMR (400
MHz, CD30D): 1.09 (s, 9H), 1.26 (s, 3H), 1.31 (s, 3H), 3.29 (m, lH),
2S 3.43 (dd, lH), 4.36 (m, 2H), 4.62 (m, lH), 6.87 (m, 2H), 7.00-7.45 (m,
lOH), 7.60 (d, lH), 8.20 (s, lH). FAB-MS: calculated for
C33H3gNsO4 569, found 570 (M+l).
Step K: (R)-a-[(2-t-Butoxycarbonylarnino-2-1nethyl-l-oxopropyl)-
amino~-N-[~2'-[[(methylamino)-carbonyl]amino][l ,l '-
biphenvll-4-yllmethvll- 1 H-indole-3-~ropanamide
The amine (25 mg, 0.04 mmol) from Step J was dissolved
in methylene chloride and methyl isocyanate (0.009 mL, 0.15 mmol)
was added to the amine. The reaction mixture was stirred together at
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room temperature for 2.5 hours then the volatiles were removed
directly under vacuum. The resultant residue was chromatographed on
silica gel using ethyl acetate/hexanes (4: 1 v/v) to afford the desired
product (23 mg, 82%). lH NMR (200 MHz, CD30D): 1.10 (s, 9H),
1.24 (s, 3H), 1.30 (s, 3H), 2.64 (d, 3H), 3.33 (ABq, 2H), 4.33 (m, 2H),
4.60 (m, lH), 6.23 (m, lH), 6.96-7.45 (m, 11H), 7.58 (d, lH), 7.68 (m,
lH), 8.20 (m, lH). FAB-MS: calculated for C3sH42N6Os 626; found
627 (M+l).
Step L: (R)~ [(2-Amino-2-methyl-1-oxopropyl)amino]-N-[[2'-
[(methylaminocarbonyl)amino] [1,1 '-biphenyl] -4-yl]methyl] -
lH-indole-3-propanamide~ trifluoroacetate
The intermediate obtained in Step K (15 mg, 0.023 mmol)
and anisole (0.01 mL, 0.09 mmol) were dissolved in methanol (0.5 mL)
15 and hexanes (0.5 mL). To this solution 0.5 mL of 9 N aqueous hydro-
chloric acid was added. The reactants were stirred at room temperature
for 30 minutes then the hexane layer was removed using a pipette. The
aqueous methanolic layer was evaporated at atmospheric pressure using
a fast stream of nitrogen at room temperature. The solid material thus
20 obtained was purified by reverse phase medium pressure liquid chroma-
tography on C8, eluting with methanol/0.1% aqueous trifluoroacetic
acid (85:15 v/v). ~his afforded 11.3 mg (0.018 mmol, 74%) of the title
compound. 1H NMR (400 MHz, CD30D): 1.38 (s, 3H), 1.56 (s, 3H),
2.66 (s, 3H), 3.18 (dd, lH), 3.33 (dd, lH), 4.35 (ABq, 2H), 4.78 (t, lH),
25 6.98-7.47 (m, lOH), 7.62 (d, lH), 7.64 (d, lH). FAB-MS: calculated
for C30H34N6o3 526; found 527 (M+l).
