CA1310792C

CA1310792C - N-heterocyclic alcohol renin inhibitors

Info

Publication number: CA1310792C
Application number: CA000528852A
Authority: CA
Inventors: Denis E. Ryono; Harold N. Weller, Iii
Original assignee: ER Squibb and Sons LLC
Current assignee: ER Squibb and Sons LLC
Priority date: 1986-02-03
Filing date: 1987-02-03
Publication date: 1992-11-24
Anticipated expiration: 2009-11-24

Abstract

ABSTRACT

N-Heterocyclic Alcohol Renin Inhibitors Compounds of the formula are disclosed wherein R1 is N-heterocyclic moiety.
These compounds intervene in the conversion of angiotensin to angiotensin II by inhibiting renin and thus are useful as antihypertensive agents.

Description

HA375a N-HETEPcOCYCLIC ALCOHOL RENIN INHIBITORS
.... . _ .

Jones et al. in WO 84/03044 disclose renin inhibiting tetra-, penta-, or hexapeptide analogues of the formula X-D-E-A-B-Z-W

where X and W are terminal groups; D, E, B and Z, of which any one or, except with reduced analogues, two may be absent, are aromatic, lipo-philic or (in the case of E) aromatic, lipophilic, or basic amino acid or amino acid analogue residues, and ~ is an analogue of a lipophilic or aromatic dipeptide residue wherein the peptide link is replaced by one to four-atom carbon or carbon-nitrogen link which as such or in hydrated form is an unhydrolyzable tetrahedral analogue of the transition state of the peptide bond as given above. In particular, A is defined as R4 Rl R2 -N - C - M C -~' ~A375a

-2~
1 3 1 079~
wherein M can b~ -C~O~.
Sz~lke et al. in European Patent ~pplication 104,041, published in March 1984, disclose renin inhibitory polypeptides including the partial sequence S
X -A- B- Z- W and X-Ph~-Hi8-A-~-Z~W

10 wherein A i~ Rl R3 R2 O.

-Ng-C~- G-~ C~- C-and G is OE~

-C8 -C~2 X is hydkog~n, protecting g~oup, or an amino acyl residue, B i~ a lipop~ilic ami~o acyl r~sidu~, and Z plu~ W ar~ an a~ino alcohol r~idu~ or Z i~
a~inoacyl and W is h~dro~y, e~t~r, amido, ~tc.
Mat~u~da et al. in U.S. Patent 4,54~,926 disclo~s renin inhibiting peptido~ of th~ for~ula N~
O C~ O Bu~
~ 2 1l 1 R - C- NH - C~ - C ~N~ C~-X

HA375a

-3-1 3 t 07q2 wherein But represents an isobutyl or sec-butyl group and X includes a group of the formula -CH(R2 )_y Gordon et al. in U.S. Patent 4,514,391 disclose hydroxy substituted peptide compounds of the formula OH R Rl o NH
C~O , which possess angiotensin converting enzyme or lS enkephalinase inhibition activity.

~ his invention is directed to new hetelocyclic alcohol containing renin inhibitors of formula I
including pharmaceutically acceptable salts thereof ~I) ll l 11 1 ~5 X ~ NH- CH - C ~ NH -CH - C -NH -CH -CH - R

0~
O O
Il ~ 11 3~ X is R6-~CH2)m~, R6-(CH2)m-C N CH C , Rlo o Il l 11 11 R6-(CH2)m-C-t NH ~CH~ C ~--q , R6-~CH2)m-O-C-, _4~ HA375a 13107Q~
o o Il 11 R6--(CH2)n~C- , R6-(CH2)m~ C-O
Il CN _ C , R6~(CH2)m-S2- or o ,, R6- ( CEI2 )m-CH--c-(CH2)m, R3, R4, R5 and R1o are independently selected from hydrogen, lower alkyl, halo substituted lower alkyl, -(CH2)n-aryl, -(CH2)n-heterocyclo, -(CH2)n-OH, -(CH2)n-O-lower alkyl, -(CH2)n-NH2, -(CH2)n-SH, -(CH2)n-S-lower alkyl, ~(CH2)n~~(CH2)g~OH, ( 2)n (CH2)g~NH2~ ~(CH2~n~S-(CH2) -OH~

ll -(CH2)n-C-OH, ~(CH2)n~S~(CH2)g~NH2, N~

30 -(CH2)n-NH-C , ~.

_5_ ~375a 1 3 1 07~2 o Il ( 2~n C NH2 ~ (CH~)n ~ J ~ I-(CH2) and -(CH2~n-cycloalkyl~
R6 and R6' are independently selected from lower alkyl, cycloalkyl, aryl and heterocyclo.
p is zero or one.
g is zero or one.
m and m' are independently selected from 5 zero and an integer from 1 to 5.
n is an integer from 1 to 5.
g is an integer from 2 to 5.

7 is CH2 O CH2 ~ or -CH2 ~ .

R8 is 2,4-dinitrophenyl, -C-O-C~2 ~ , -52 ~ CH3 ~ or -CH2 O-CH

~0 -6- HA375a Rl is a fully saturated, partially saturated, or unsaturated monocyclic N-heterocyclic ring of 5 or 6 atoms containing at least one N atom or a ~icyclic ring in which such N-heterocyclic ring is fused to a benzene rin~. The N-heterocyclic ring can also include an O or S atom or up to three additional N atoms. The N-heterocyclic ring is attached to -CH- by way of an available carbon OH
atom. An available N atom in the N-heterocyclic ring can be substituted with an N-protecting group-such as -CH2-0-CH2 ~ , -S2 ~ H3 ~

or 2,4-dinitrophenyl, or lower alkyl, -(CH2)n ~ , or -(CH2)n-cycloalkyl. Similarly, an available C
atom in the monocyclic ring or an avaliable C atom in the benzene portion of the bicyclic ring can be substituted with lower alkyl, ~(CH2)n~> ' or -(CH2)n-cycloalkyl.
Preferred N-heterocyclic rings are lR2 lR2 ~ ~ Rg, N ~ 9 ' N ~ 9 R

r~ ~ 2 Rg HA375a ~7 1 3 1 0792 ~S ~o N $1 N ~J
R~ ~'<9 R9 ~ N ~ Rg ' --N~ ~ 9 _~Rg , ~ Rg ~ ~Rg T R~ and ~N~ Rg -2 5 CH2 0 CH2~ , -S02~ c~3 2,4-dinitrophenyl, hydrogen, lower alkyl, -(CH2) or -(CH2)n-cycloalkyl.
Rg is hydrogen, lower alkyl, -(CH2) or -(CH2)n-cycloalkyl.

-8- HA375a ~ 3 1 0 7 9 2 ,--~ N- represents a heterocyclic ring of the S ~ormula ~ (CH2)~\

Y N-(CH~)b wherein Y is -CH2, O, S, or N-Rg, a is an integer from 1 to 4, and b is an integer from 1 to 4 provided that the sum of a plus b is an integer ~rom 2 to 5 and such heterocyclic rings wherein one carbon atom has a substitu~nt selected from lower alkyl, lower alkoxy, lower alkylthio, halo, CF3, nitro, or ~ydroxy.

~0 This invention in its broadest aspects relates to the compounds of formula I above, to compositions and the method of using such compounds as antihypertensive agents, and intermediates useful in the preparation of such compounds.
The term lower alkyl used in defining various symbols refers to straight or branched chain radicals having up to seven carbons.
Similarly, the terms lower alkoxy and lower alkylthio refer to such lower alkyl groups attached to an oxygen or sulfur. ~he preferred lowar alkyl groups are straight or branched chain o~ 1 to 5 carbons.

_g_ HA375a 1 31 07~2 The term cycloalkyl refers to saturated rings of 4 to 7 carbon atoms with cyclopen~yl and cyclohexyl being most preferred.
The term halogen refers to chloro, bromo and fluoro.
The term halo substituted lower alkyl refers to such lower alkyl groups described above in which one or more hydrogens have been replaced by chloro, bromo or fluoro groups such as trifluoro-methyl, which is preferred, pentafluoroethyl,2,2,2-trichloroethyl, chloromethyl, bromomethyl, --etc.
The term aryl refers to phenyl, l-naphthyl, 2-naphthyl, mono substituted phenyl, l-naphthyl, lS or 2-naphthyl wherein said substituen~ is lower alkyl of 1 to 4 carbons, lower alkylthio of 1 to 4 carbons, lower alkoxy of 1 to 4 carbons, halogen, hydroxy, amino, -NH-alkyl wherein alkyl is of 1 to

4 carbons, or -N(alkyl)2 wherein alkyl is of 1 to 4 carbons, di or tri substituted phenyl, l-naphthyl or 2-naphthyl wherein said substituents are selected from methyl, methoxy, methylthio, halo~en, and hydroxy.
The term heterocyclo refers to fully saturated or unsatuxated rings of 5 or 6 atoms containing one or two 0 and S atoms and/or-one to four N atoms provided that the total number of hetero atoms in the ring is 4 or less. The hetero ring is attached by way o an available carbon atom.
Preferred hetero groups include 2- and 3-thienyl, 2- and 3-furyl, 2-, 3- and 4-pyridyl. The term hetero also includes bicyclic rings wherein HA375a -lO- I 3 1 0792 the five or six membered ring containing 0, s and N atoms as defined above is fused to a benzene ring. The preferred bicyclic ring is benzimida201yl.
The compounds of formula I wherein o Il 6 ~ H2)m C
can be prepared by coupling an alcohol of the formula (II) OH

with a peptide of the formula ~III) R6-(CH2)m~O~C-~ N~ -CH - C~-p NH- CH - COOH

This reaction is preferably performed in a solvent such as dimethylformamide and in the presence of hydroxybenzotria201e, N-methylmorpholine, and a coupling agent such as dicyclohexylcarbodiimide.
The corresponding compounds of formula I
wherein p is zero can be prepared by coupling the alcohol of formula II with the amino acid of the formula -11- HA375a 1 3 1 0 ~q~
(IV~

Il I
R6-(cH2)m-o-c-NH-cH -COOH
to yield the products of the formula (V) Rl R3 Il l 11 1 R6(C~)m-O-C-NH-CH-C -NH-CH -CH-R

OH

When R6-(CH2)m- is t-butyl or benzyl, lS then t~e product of formula V can be treated so as to remove the t-butoxycarbonyl or ben2yloxycarbonyl group such as by the use of anhydrous hydrochloric acid when R6 is t-butyl to yield the amine of the formula (VI) Il H2N-C~--C-- N~--CH--CH-Rl I

~5 OH

Coupling with the acylated amino acid of the formula (VII) O R
~.
R6-(CH2) -O-C -NH -CH -COOH

yialds ~he products of formula I wherein p iS one.

HA375a ~ 3 1 ~7q2 The compounds of formula I wherein X is o Il other than R6-(CH2)m-O-C-s can b~ prepared by treating the product of formula I wherein X is O o -C -o-c(cH3)3 or -C O C~2 ~
to remove the t-butoxycarbonyl or benzyloxycarbonyl group and yield the intermediates of the formula _-(VIII) R5 1I R4 11 l3 H2N-CH--C--NH--CH--C--N~--CH~ CH-R

OH

The amine of form~la VIII or formula VI is treated with the halide of the formula (IX) R6- ( CH2 )m-halo particularly where halo is Br to give the products of formula I wherein X is R6-(CH2)m~.
~he compounds of formula I wherein X is o R6-O-(CH2)n-C- can be prepared by treating ~le amine of formula VIII or VI with the acid chloride -13- HA375a of the formula (X) o Il ~6 (CH2)n-c-Cl in the presence of triethylamine.
The compounds of formula I wherein X is R6-(CH2)m-S02- can be prepared by treating the amine of formula VIII or VI with the substituted sulfonyl chloride of the formula (XI) 6 ( 2)m 2 ~he compounds of formula I wherein X is o Il R6-(CH2)m-C- can be prepared by treating the amine of formula VIII or VI with the acid chloride of the formula (XII) O
Il R6- ( CH2 )m-C-Cl ~5 in ~he presence of triethylamine. Alternatively, these compounds can also be prepared by coupling the carboxylic acid of the fo~ula (XIII) 3~ 0 R6-(CH2)m-C-OH

~ 3 1 0792 HA375a to the amine of formula VI or VIII in the presence of a coupling agent sucn as dicyclohexylcarbo~
diimide and 1-hydroxybenzotriazole hydrate.
~he compounds of formula I wherein X is Rl o O
Il l 11 R6- ( CH2 )m~C~ (NH-CH--C ~

and q is one can be prepared by acylating the amino acid of the formula (XIV) -.
RllO
H2~CH - C - OH
with the acid chloride of formula XII in the presence of base such as sodium hydroxide (i.e. at a pH of about 8) and in a solvent such as tetra-hydrofuran and water to give the acylated amino ~O acid of the formula (XV) Rlo Il l 11 R~-~C~2) -C-N~-CH - C - OH
~5 The amino acid of formula XV is then coupled to the amine of formula VI or VIII in the presence of dicyclohexylcarbodiimide and l-hydroxybenzo-triazole hydrate to give the desired compounds of formula I.
The compounds of the ~ormula I wherein X is o Il R~-(CH2)m-NH-C- and p is one can be prepared by coupling an amino acid of the formula HA375a (XVI) Il l 11 6 (CH2)m-N~-c-NH-cH--C--OH
to the amine of formula VI in the presence of a coupling agent such as dicyclohexylcarbodiimide and l-hydroxybenzotriazole hydrate.
Similarly, the compounds of formula I
wherein X is Q
Il . . .
R6- ( CH2 )m-NH-C-and p is zero can be prepared by coupling an amino acid of the formula (XVII) R6-(CH2)m-NH-C-NH--CH--C--OH
to an alcohol of formula II in the presence of a coupling agent such as dicyclohexylcarbodiimide and l-hydroxybenzotriazole hydrate.
The compounds of formula I wherein X is O

CN_ C and p is one can be prepared by coupling an amino acid of the formula (XVIII) O R5 O ~.
Il l 11 CN _ C _NH C~_ C _ OH

to the amine of formula VI in the presence of a coupling agent such as dicyclohexylcarbodiimide 1 3 1 07q2 HA375a and l-hydroxyben20triazole hydrate.
Similarly, the compounds of formula I

wherein X is C N - C- and p is zero can be prepared by coupling an amino acid of the formula ~XIX) Il 1 11 .
CN_ C_ NH ~ CH - C -O~H

to the alcohol of formula II in the presence of a coupling agent such as dicyclohexylcarbodiimide and 1-hydroxybenzotriazole hydrate.
The amino acid intermediates of formulas XVI, XVII, XVIII, and XIX can be prepared by treating an amine R6-(CH2)m-NH2 with phosgene and N-methyl ~0 morpholine followed by reaction with an amino acid methyl ester hydrochloride salt of the formula (~) l 11 H2N-CH - C - 0 -CH3 ~Cl or o the formula (XXI) 3a 1 11 ~.
H2N-CH - C -0 -CH3 ~ HCl in the presence of N-methyl morpholine. Removal of the methyl ester group by treatment with aqueous 3~ sodium hydroxide gives the desired intermediate.

HA375a The products of formula I wherein X is R5-(CH2)m CH -C-I

R' can be prepared by coupling the carboxylic acid of the formula _~
(XXII) o Il R5-(CH2)m CH- C -o~

(CH2)~, I

R'6 to the amine of formula VI or VIII in the presence of dicyclohexylcarbodiimide and l-hydroxybenzo-triazole hydrate. Alternatively, the acid of formula XXII can be converted to the acid chloride and this acid chloride can then be coupled to the amine of formula VI or VIII in the presence of triethylamine and tetrahydrofuran or water and sodium bicarbonate.

13~07q2 HA375a The compounds of formula I wherein X is O o Il ~I 11 R6-(C~2)m- C - N - C~-C-can be prepared by acylating proline with the acid chloride of formula XII in the presence of base such as sodium hydroxide, i.e., a pH of about 8~
and a solvent mixture of tetrahydrofuran and water to giv~ ..
(XXIII) O o R6-(CH~)m C N - CH-C-OH

The acylated amino acid of formula XXIII is then coupled to the amine of formula VIII or VI in the presence of a coupling agent such as dicyclo-hexylcarbodiimide and l-hydroxybenzotriazole hydr~te.
~ he alcohol of formula II can be prepared by treating the N-heterocyclic starting material of the formula H-Rl with n-butyl lithium to yield the lithium compound of the formula ~YIV) Li-R

.

HA375al 3 1 0 7, 2 This lithium N-heterocyclic compound is then reacted with the aldehyde o the formula ~YV) Prot-NH- CH -CH
wherein Prot is an amino protecting group such as t-butoxycarbonyl. Removal of the t-butoxycarbonyl group such as by treatment with hydrochloric acid 1~ givas the alcohol of formula II.
The aldehyde of formula XXV is prepared by .
treating the N-protected ~-amino acid ester of the formula (~XVI) l 11 Prot-NH-CH- C- O-alkyl with lithium borohydride to give the alcohol of the formula ~YVII) I

Prot -NH- CH - CH2 OH
Traatment of the alcohol of formula XXVII with ~5 pyridine-sulfur trioxide complex or with periodinane reagent [see Dess et al., J. Org.
Che~., Vol~ 48, p. 5155-4156 (1983)] gives the d~sired aldehyde.
In the above reactions, if any of R3, R4, R5 ~nd Rlo are -(CH2)n-aryl wherein aryl is phenyl, l-naphthyl, 2-naphthyl substituted with one or 1 3 ~ 0792 HA375a more hydroxy or amino groups, -(CH2)n-heterocyclo wherein heterocyclo is an imidazolyl, -(CH2)n~NH2, ( 2)n SH, (CH2)n~OH, -(CH2) -NH-C or o Il -(CH2)n-C-OH then the hydroxyl, amino, imidazolyl, mercaptan, carboxyl, or guanidinyl function should be protected during the reaction. Suitable protect-ing groups include benzyloxycarbonyl, t-butoxycarbonyl, benzyl, benzhydryl, trityl, etc., and nitro in the case of guanidinyl. The protecting group is removed by hydrogenation, treatment with acid, or by other known means following completion of the reaction.
The various peptide intermediates employed in above procedures are known in the literature or can be read~ly prepared by~known methods. See for example, The Peptides, Volume 1, "Major Methods Of Peptide Bond Formation", Academic Press (1979).
Preferred compounds of this invention axe those of formula I wherein:
o ll X is R6 (C~2)m C

Rlo R6 (CH2)m N~ C~ , R6-(CH2)m~~C~

1 3 1 ~7q2 -21- HA375a o o R6- ( CH2 )m-N}I-C- CN_C_ or O
Il 6 ( H2)m ~CH C
(CIH2~m~

R~ and R6' are independently selected from straight or branched chain lower alkyl of up to 5 carbons, cycloalkyl of 4 to 6 carbons, phenyl, l-naphthyl, and 2-naphthyl.
m and m' are independently selected from zero, one and two.
C CN_ CN_ ~o ~ /~ ~ ~
JW O N- S ~ N- HN N
H3C-N ~N-~s Especially preferred are the compounds wherein X is O
3~
(H3C)3C-O-C-- ~ ~ ~ CH27~ CH-C- ,, 1 3 1 07~2 O o l } c- , (H3C)3-C-CH2-C-o (H3C)3-C-NH-C-o o o (H3C)3-C-C- , C-N~-CH--C-( CH2 ) 4 O O
CN_C_ O N-C-\ -f~
or H3C-N N-~ 9 N ~ 9 ~3Rg ~R ~ N _ -R2 1 3 1 07~2 HA375a Rg ~ ~ ~ Rg , Rg R2 ~ Rg ~ Rg , especially ~NI ~ ' ~ N

2 is CH2 CH2 ~ , hydrogen, straight or branched chain lower alkyl of up to 5 carbons, or -(CH2)n ~ wherein n is an integer from 1 - to 3.

HA375a -24~

Rg is hydrogen, straight or branched chain lower alkyl of up to 5 carbons, or -~CH2) S wherein n is an integer from 1 to 3.
R3 is straight or branched chain lower al~yl of 3 to 5 carbons, -(CH2)n-cyclopentyl, -(CH2)n-cyclohexyl, or -(CH2)n ~
wherein n is an integer from 1 to 3, especially -.

-CH2 ~ or -CH2CH(CH3)2 R4 is hydrogen, straight or branched chain lower alkyl of up to 5 carbons, -(CH2)4-NII2 , r fJ 2 ~ ~N-cH2-o-cH2 ~ , -CX~ ~ ~ -CH2 ~ ~ _cu2 ~ OH , -C~ -C:~2~N , -Cn2~ ' 1 3 ~ ~) 7 HA375a -(C~2) ~ '` or -''^2 ~ ~2 ~2 especially 2 ~ ~J

R5 is strai~ht or branched chain lower alkyl of up to 5 carbons, -CH2 ~ , -(C:.2) ~ , -CH2 ~ - OCH3 -CH2-~-naphthyl), -CH2-(~-naphthyl), -CH2 ~ OH, -CH2-cyclopentyl, -CH2-cyclohexyl~ C:' ~

-c-~2~ ,3 ~ 2 ~ , especially benzyl.

y 1 31 07~2 HA375a Rlo is (CH2)4 NH2 The compound of foxmula I form salts with a variety of inorganic and organic acids. The non-toxic pharmaceutically acceptable salts are pre-ferred, although other salts are also useful in isoiating or purifying the product. Such pharma-ceutically acceptable salts include those formed with hydrochloric acid, methanesulfonic acid, sulfuric acid, acetic acid, maleic acid, etc. The ~alts are obtained by reacting the product with an~
equivalent amount of the acid in a medium in which the salt precipitates.
The compounds o formula I contain asymmetric centers when any or all of R3, R4, R5 and Rlo are other than hydrogen and at the carbon to which the -OH
group i~ attached. Thus, the compounds of formula I
can axist in diastereoisomeric forms or in mixtures thereof. The above described processes can utilize racemates, enantiomers or diastereomers as starting materials. When diastereomeric products are prepared, they can be separated by conventional chromatographic or fractional crystallization methods.
The compounds of formula I, and the pharma-~5 ceutically acceptable salts thereof, are anti-hypertensive agents. They inhibit the conversion o~ angiotensinogen to angiotensin I and therefore, are useful in reducing or relieving angiotensin r~lated hypertension. The action of the enzyme renin on angiotensinogen, a pseudoglobulin in blood plasma, produces angiotensin I.

~ 3 ~ 0792 HA375a Angiotensin I is converted by angiotensin converting enzyme (ACE) to angiotensin II. The latter is an active pressor substance which has been implicated as the causative agent in several S forms of hypertension in various mammalian species, e.~., humans. The compounds of this invention intervene in the angiotensinogen ~
(renin) ~ angiotensin I ~ tACE) ~ angiotensin II
sequence by inhibiting renin and reducing or eliminating the formation of the pressor substance angiotensin II. Thus by the administration of a _.
composition containing one (or a combination~ of the compounds of this invention, angiotensin dependent hypertension in a species of mammal (e.g., humans) suffering therefrom is alleviated.
A single dose, or preferably two to four divided d~ily doses, provided on a basis of about 100 to 1000 mg., preferably about 250 to 500 m~. per kg. of body weight per day is appropriate to reduce blood ~0 pressure. ~he substance is preferably administered orally, but parenteral routes such as the subcutaneous, intramuscular, intraveneous or intraperitoneal routes can also be employed.
The compounds of this invention can also be ~S formulated in combination with a diuretic for the treatment of hypertension.
~ combination product comprising a compound of this invention and a diuretic can be administered in an efective amount which comprises a total daily dosage of about 1000 to ~000 mg~, preferably about 3000 to 4000 mg. of a I 3 1 07~2 HA375a compound of this invention, and about 15 to 300 mg., preferably about 15 to 200 mg. of the diuretic, to a mammalian species in need thereof.
Exemplary of the diuretics contemplated for use in combination with a compound of this invention are the thiazide diuretics, e.g., chlorothiazide, hydrochlorothiazide, flumethiazide, hydroflu-methiazide, bendroflumethiazide, methyclothiazide, trichloromethiazide, polythiazide or benzthiazide as well as ethacrynic acid, ticrynafen, chlorthalidone, furosemide, mu~olimine, bumetanide, triamterene, amiloride and spironolactone and salts o~ such compounds.
The compounds of formula I can be formulated for use in the reduction of blood pressure in compositions such as tablets, capsules or elixirs for oral administration or in sterile solutions or suspensions for parenteral administration.
About 100 to 500 mg. of a compound of formula I is compounded with physiologically acceptable vehicle, carrier, excipient, binder, preservative, stabilizer, flavor, etc., in a unit dosage form as called for by accepted pharmaceutical practice.
The amount of active substance in these ~5 compositions or preparations is such that a suitable dosage in the range indicated is obtained.
The following examples are illustrative of the invention. All temperatures are given in degrees centigrade.

HA37sa 7 3 ~ ~192 Example N -[N-[(1,1-Dimethylethoxy~carbonylL~L~ y~
alanyl]-N-[(S)-l-~hydroxy(lH-imidazol~2-yl~methyll-3-methylbutYl~-L-histidinamide, ace~lc acid solvate a) N-[(l,l-Dimethylethoxy)carbonyll-L-leucinal Thionyl chloride (50.2 ml., 691 mmole) is added dropwise over a period of 20 minutes to a stirred (ice-cold) suspension of L-leucine (72.05 g., 550 mmole) in absolute ethanol. After the addition is completed, the ice-bath is removed and the reaction mixture is stirred at room temperature for one hour. It is then refluxed for }ive hours on a steam bath. It is then concentrated in vacuo and diluted with ether. The -~eparated crystals are filtered to give 99.9 g. of L-leucine, ethyl ester, monohydrochloride; m.p.
~130) 135-137; [~]22 = +14.7 ~c = 2.3, methanol), A solution of L-leucine, ethyl ester, mono-hydrochloride (46.85 g., 239.4 mmole) in 75%
ethanol (500 ml.) is added to a vigorously stirred solution of sodium borohydride (35 g., 907 mmole) in 75% ethanol (500 ml.) over a period of thirty minutes. After the addition is completed, the ~5 reaction mixture is refluxed for 3 hours. Ethanol is removed ln vacuo. The agueous solution is extracted with ethyl acetate. The aqueous solution is again extracted with ethyl acetate after saturating with sodium chloride. The combined ethyl acetate solution after washing with saturated sodium chloride solution is evaporated to HA375a 1 3 1 0 7 9 2 give 20.8 g. of crude (S~ 2-amino-4-methyl~l-pentanol.
Di-tert-butyl dicarbonate (38.73 g., 177.5 mmole) is added to a stirred (ice-bath) solution of (S)-2-amino-4-methyl-1-pentanol (20.8 g., 177.5 mmole) in tetrahydrofuran (340 ml.). After stirring in an ice-bath for 15 minutes, the reaction mixture is stirred at room temperature for 3 hours. The tetrahydrofuran is removed ln vacuo. This crude product (~0.53 g.) is combined with 36.25 g. from a previous run and the entire ~lount is chromatographed over silica gel (800 g;)_~
using the solvent system ethyl acetate:hexane (1:1) to yield 72.6 g. of (S)-2-[[(l,l-dimethyl-ethoxy)carbonyl]amino]-4-methyl-1-pentanol as an oil; [~]D2 = -26.2 (c = 1.5, methanol).
Trie~hylamine (63 ml., 450 mmole), dimethyl-sulfoxide (63.9 ml., 900 mmole), and pyridine-sulur trioxide complex (71.6 g., 450 mmole) ~re added to a stirred (room temperature) solution of (S)-2-[[(1,1-dimethylethoxy)carbonyl]amino]-4 methyl-l-pentanol (32.59 g., 150 mmole) in methylene chloride (750 ml.). After stirring the reaction mixture for 15 minutes, it is evaporated in vacuo at room temperature. Ice-water (450 ml.) ~5 is added to the residue which is then extracted with ether. The e~her extract is successively washed with 10% citric acid, water, saturated sodium bicarbonate, and water. The ether extxact on evaporat,on gives 19.73 g. of N-[(l,l-dimethyl-ethoxy)carbonyl~-L-leucinal as an oil; ~]22 = _50O
(c = 4.6, methanol).

~ -31~ HA37sa 1 3 1 0 7 q 2 b) ~-~(S)-1-Amino-3-methylbutyll-1-[(phenyl-methoxy)methyl~-lH-imidazole-2-methanol, dihydrochloride 2.5 M n-Butyllithium solution in hexane (4.6 ml., 11.5 mmole) is added to a solu~ion of l-[(phenylmethoxy)methyl]-lH-imidazole (2.06 g., 10.94 mmole; prepared as described by Brown et al., J.Chem.Soc. Perkin Trans. I, l9B2, p. 1553) in tetrahydrofuran (35 ml.) at -70 under argon.
After 45 minutes at -70, an orange colored solution results. A~ this point, a solution of --N-[(1,1-dimethyleth~oy)carbonyl]-L-leucinal (1.18 g., 5.47 mmole) in tetrahydrofuran (7 ml.) is carefully added over a period of 2 - 3 minutes.
The reaction is kept at -70 for one hour, then at 0 for 15 minutes, and then is quenched by the addition of saturated ammonium chloride (6 ml.).
The reaction mixture is diluted with ether and rinsed with several portions of water and brine, and dried over magnesium sulfate. Concentration ln vacuo gives 3.13 g. of crude product. Flash chromatography on silica gel (120 g. of Whatman LPS-1) elutiny with petroleum ether-acetone t4:1) gives 261 mg. of ~-[(S)-l-[[(l,l-dimethylethoxy)-carbonyl]amino]-3-methylbutyl]-1-[(phenylmethoxy) methyl]-lH-imidazole-2-methanol, fast moving isomer; [a]22 = +5.8 (c - 0.5, chloroform), 564 mg. of a-[(S)-l-[[(1,1-dimethylethoxy)-carbonyl]amino]-3-methylbutyl]-l~[(phenylmethoxy)-methyl]-lH-imidazole-2-methanol, slow moving isomer; [a]22 - +12.2 (c = 0.5, chloroform), and 625 mg. of a mixture fraction for a total -32- HA375a yield of 1.45 g. of product; TLC (silica gel;
petroleum ether:acetone, 3:1) Rf = 0.39, 0.13.
Anal. calc'd. for C22H3~N304:
C, 65.48; H, 8.24; N, 10.41 Found: C, 65.05; H, 8.20; N, 10.21 (fast moving isomer) C, 65.26; H, 8.37; N, 10.29 (slow moving isomer) A (1:1) isomeric mixture of the above fast and slow moving isomers (493 mg., 1.22 mmole) is dissolved in 10 ml. of ethyl acetate and cooled in_.
an ice-water bath under argon. The solution is saturated with dry hydrochloric acid and stirred cold for one hour, and then concentrated 1n vacuo to give 420 mg. of crude product. Chromatography on a 30 x 350 mm. HP-20 column gradient eluted from 350 ml. of 9:1 to 1:9, 0.01 N aqueous hydro-chloric acid:acetonitrile at 9 ml.~2 min./fraction yields 402 mg. of a-[(S)-1-amino-3-methylbutyl-1-[(phenylmethoxy)methyl]-lH-imidazole-2-methanol, dihydrochloride; m.p. 98-117; [a]22 = _0 3O
(c = 1, methanol). TLC (silica gel; n-butanol:
pyridine:acetic acid:water, 4:1:1:1) Rf = 0.78.
Anal. calc'd for C17H25N3O2 2HC1 1.7 H2O:
~5 C, 50.17; H, 7.53; N, 10.33; Cl, 17.42 Found: C, 50.13; H, 7.54; N, 10.29; C1, 17.58.
c) N-~N-[(l,1-Dimethylethoxy)caxbonyl~-L-phenyl-alanvll-l'-[(phenylmethoxy)methyl]-L-h-istldine Thionyl chloride (27.2 ml., 375 mmole) is added in drops to a stirred solution in an ice-1 3 1 07q2 _33_ HA375a bath of L-histidine (38.75 g., 2~0 mmole) in methanol (500 ml.). After 15 minutes the ice-bath is removed and the reaction mixture is stirred at room temperature for one hour. After refluxing for 48 hours, it is concentrated ln vacuo. The separated crystals are filtered using methanol for washings to 48.93 g. of L-histidine, methyl ester, dihydrochloride. The methanolic solution on dilution with ether affords an additional 10 g. of product; m.p. 208 - 209; [~]2D2 = +10.1 (c = 1.8, water).
Triethylamine (28 ml., 200 ml.) and di-tert-butyl dicarbonate (48 g., 220 mmole) are added to a suspension of L-histidine, methyl ester (24.~ g., 100 mmole) in methanol (80 ml.). After 3.5 hours, the mixture is filtered and the methanolic solution is concentrated ln vacuo. The residue is taken into chloroform and washed with 10~ citric acid. The crude product on crystallization-from isopropyl ether affords 23.1 g. of N,l'-bis~(l,1-dimethylethoxy)carbonyl]-L-histidine, methyl ester;
m.p. (62) 88 - 95; [~]2D2 = +25 4 (c = 1 1 carbon tetrachloride).
Benzylchloromethyl ether (11.6 ml., 83.6 mmole) is added to a solution of N,1'-bis[~1,1-dimethyl-ethoxy)carbonyl]-L-histidine, methyl ester (24.7 g., 66.9 mmole) in dry methylene chloride (156 ml.) and the reaction mixture is stirred at room temperature for 5 hours. After concsntrating ~n vacuo and on dissolution in ethyl acetate 17.85 g.
of N-[~1,1-dimethylethoxy)carbonyl]-1'-[~phenyl-methoxy)methyl]-L-histidine, methyl ester, mono HA375a 1 3 1 07~2 hydrochloride crystallizes out; m.p. (148) 152 -153; ~]D = -19.5~ (c = 1.8, methanol). This methyl ester product is dissolved in hydrogen chloride in acetic acid solution (60 ml., 1.5 N
and kept at room temperature for 15 minutes. It is then evaporated in vacuo and the residue is dissolved in hot isopropanol. After cooling, the separated crystals are filtered to yield 7.08 g.
of l-[(phenylmethoxy)methyl]-L-histidine, methyl ester, dihydrochloride; m.p. (170) 173 - 17~.
l-[(Phenylmethoxy)methyl]-L-histidine, methyl-ester, dihydrochloride (1.79 g., 4.94 mmole), l-hydroxybenzotriazole (0.756 g., 4.94 mmole), and N-[(1,1-dimethylethoxy)carbonyl]-L phenylalanine (1.31 g., ~.94 mmole) are dissolved in dimethyl-formamide (16 ml.). While stirring the above solution in an ice-bath, dicyclohexylcarbodiimide (1.02 g., 4.94 mmole) and N,N-diisopropylethylamine (1.72 ml., 10 mmole) are added. After 3 hours the ice-bath is removed and the reaction mixture is stirred at room temperature overnight. It is then concentrated to dryness and the residue is tritura-ted with ethyl acetate. The saparated urea is filtered off. The ethyl acetate solution is washed with saturated sodium bicarbonate and then it is evaporated. The residue upo~ crystallization from ethyl acetate gives 1.97 g. of N-[N-[(l,1-dimethylethoxy)carbonyl]-L-phenylalanyl]~
~(phenylmethoxy)meth~l]-L-histidine, methyl ester;
m.p. (165) 166 - 168.

13107~2 HA375a N-[N-[~l,1-Dimethylethoxy)carbonyl]-L-phenylalanyl]-l'-[(phenylmethoxy)methyl]-L-histidine, methyl ester (4.5 g., 8.4 mmole) is dissolved in hot methanol (25 ml.). After cooling to room temperature aqueous sodium hydroxide solution t9.24 ml., lN) is added and the mixture is stirred at room temperature for 3 hours. It is then concentrated in vacuo and water (60 ml.) is added to the residue. After cooling the aqueous 1~ ~olution in an ice-bath, it is acidified to pH 4~5 u~ing aqueous hydrochloric acid. It is then --extracted with ethyl acetate to yield 3.95 g. of crystalline N-[N-[~1,l-dimethylethoxy)carbonyl]-L-phenylalanyl]~ [(phenylmethoxy)methyl]-L-histidine; m.p. 193 - 194; [~]22 = -4.8 (c = 1.1, dimethylformamidè).
d) N -~N-[(l,l-Dimethylethoxy~carbonyl]-L-phenyl-alanyll-N- r (s,-1- ~hydroxy[l-[(phenYlmethoxY)methvl~-lH-imidazol-2-yllmethyl~-3-methylbutyl]-3'-[(phenvl-~0 methoxy)methyl]-L-histidinamide N-Methylmorpholine (154 mg., 1.52 mmole~ is added to a mixture of N-[N-[(l,l-dimethylethoxy)-carbonyl]-L-phenylalanyl]-l'-[(phenylmethoxy)methyl]-L-hi~tidine (397 mg., 0.759 mmole), ~-[(S)-l-amino-3-~S methylbutyl]-l-[(phenylmethoxy)methyl]~lH-imidazole-2-methanol, dihydrochloride (309 mg., 0.759 mmole), and l-hydroxybenæotriazole hydrate (116 mg., 0.759 mmole) in dimethylformamide (5 ml.) cooled in an ice-water bath under argon followed by the 30 addition of dicyclohexylcarbodiimide (157 mg., 1 3 1 07q2 HA375a 0.759 mmole). The reaction mixture is kept cold for 2 hours and then refigerated overnight. The reaction mixture is then diluted with ethyl acetate (30 ml.), chilled for 20 minutes, and then filtered. The filtrate is further diluted with ether and the organic solution is rinsed with two portions of water (15 ml.), saturated sodium bicarbonate solution (15 ml.), and brine, dried over magnesium sulfate, and concentrated ln vacuo to give 615 mg. of crude product. Two flash chromatographies on silica gel (LPS-l) eluting .-with chloroform:methanol:ammonia (25:2:0.05) gives 385 mg. of N2-~N~ dimethylethoxy)carbonyl]-L-phenylalanyl]-N-[(S)-l-[hydroxy[l-[(phenylmethoxy)-methyl]-lH-imidazol-2-yl]methyl]-3-methylbutyl]-3'-[(phenylmethoxy)methyl]-L-histidinamide; m.p.
69-84; [~]D = -14.3 (c = 1, methanol). TLC
(silica gel; chloroform:methanol:ammonia, 25:2:0.05) Rf = 0.2, 0.26.
2 Anal- calc d- for C45H57N7O7 05 H2O
C, 66.15; H, 7.16; N, 12.00 Found: C, 66.20; H, 7.13; N, 11.73.
e) N2-[N-[(l,1-Dimethylethoxy)carbonyl]-L-phenYl-alanyl]-N-[(S~ hydroxy(lH-imidazol-_-yl)methyl]-3-methylbutyll-L-histidinamide, acetate A solution of the product from part (d) (337 mg., 0.413 mmole) in a mixture of methanol (16 ml.), water (3.1 ml.), and lN aqueous hydrochloric acid (0.908 ml.) is stirred under an atmosphere of hydrogen (balloon) in the presence o ?0% palladium hydroxide on carbon catalyst 1 3 1 37 9 ~

HA375a (80 mg.). After 17 hours, the reaction mixture is filtered, concentrated ln vacuo, and flash chroma-tographed on 36 g. of silica gel (LPS-l) eluting with chloroform:methanol:water:acetic acid (80:20:~.5:1). Pooliny of the product containing fractions gives 187 mg. of N2-[N-[(l,l-dimethyl-ethoxy)carbonyl]-L-phenylalanyl]-N-[(S)-1-[hydroxy(lH-imidazol-2-yl)methyl]-3-methylbutyl]-L-histidinamide, acetate; m.p. 90-118;
[~]D~ = -30.8~ (c = 0.5, methanol).
TLC`(silica gel; chloroform:methanol:water:acetic --acid, 90:20:2.5:1) Rf = 0.14 and 0.17.
Anal. calc'd. for C29H41N7O5 2.4 C2 ~ 2 C, 54.2S; X, 7.36; N, 13.11 Found: C, 54.16; H, 7.01; N. 13.07.
Exam~le 2 N -~N-[(l,l-Dimethylethoxy)carbonyl]-L-~henyl-alanyl]-N-~(S)-1-[(S)-hYdroxy(lH-imidazol-2-yl)-methyll-3-me_h~vlbutyl~-L-histidinamide, acetic ?O acid solvate "
a) NC-[N-L51,1-Dimethylethoxv)carbonyl]-L-~henyl-alanyl~-N-[(S)-l-~(S~-hydroxy~ (phenylmethoxy) m-thyl]-lH-imidazol-2-yllmethyll-3-methylbutyl~
3'- r (~henvl--m-ethoxy)methyl]-L-histidinamide ~5 A solution of ~-[(S)-1-[[(1,l-dimethylethoxy)-c~rbonyl]amino]-3-methylbutyl]-1-[~phenylmethoxy)-mathyl]-lH-imidazole-2-methanol, fast moving isomer, from Example l(b), 320 mg., 0.79 mmole) in ethyl acatate (7 ml.) is cooled in an ice-water bath and saturated with gaseous hydrogen chloride. After remaining in the bath for 15 minutes, the bath is ~ 3 1 37q2 HA375a removed and the mixture is allowed to stand at ambient temperature in a stoppered flask, for 65 minutes. The solution is concentrated 1n vacuo to give 300 mg. of a solid white powder residue of ~-[(S)-1-amino-3-methylbutyl]-1-[(phenylmethoxy)methyl]-lH-imida~ole-2(S)-methanol, dihydrochloride; m.p. 70-100~.
The above powder (270 mg., 0.66 mmole) is dissolved in dry dimethylformamide (5 ml.).
1-H~droxybenzotriazole hydrate (100 mg., O.66 mmole), N-[N-[(1,l-dimethylethoxy)carbonyl]- ' L-phenylalanyl]~ [(phenylmethoxy)methyl]-L-histidine ~342 mg., 0.66 mmole) and N-methyl-morpholine (1~7 mg., 1.38 mmole) are added and the mixture is cooled in an ice-water bath and treated with dic~clohexylcarbodiimide (149 mg., 0.66 mmole). The mixture is stirred, in a stoppered flask, in the cold for 15 minutes and then refrigerated for 18 hours. After diluting to a volume of 35 ml. with ethyl acetate, the mixture is filtered to remove insolubles and 35 ml. of ether is added. After washing with water (2 x 15 ml.), saturated sodium bicarbonate solution ~15 ml.) and brine, the mixture is dried ~5 over magnesium sulfate and concentrated ln vacuo to give 600 mg. of crude product as a viscous oil.
This residue is chromatographed on silica gel (LPSl, 90 g.) eluting with chloroform:methanol:
con-entrated ammonium hydroxide (30:~:0.05) to give 200 mg. of N2-[N-[(l,l-dimethylethoxy)-carbonyl]-L-phenylalanyl]-N~[(S)-l-[(S)-hydroxy-HA375a [1-[(phenylmethoxy)methyl]-lH-imidazol-2-yl]-methyl]-3-methylbutyl]~3'-[(phenylmethoxy)methyl]-L-histidinamide as a glassy solid;
m.p. 70-100; [a]D = 27.5 (10 mg./ml. of methanol). TLC (silica gel; chloroform:methanol:
concentrated ammonium hydroxide, 30:2:0.05) Rf = 0.10.
Ana a 45 57 7 7 C, 65.86; H, 7.17; N, 11.95 Found: C, 65.86; H, 7.12; N, 11.76.
b) `N2-[N-[(1,1-DimethylethoxY)carbo~y~J-L-phenyl~
alanyl]-N-r(S)-l-[~S~-hydroxY(lH-imidazol-2-yl)methyll-3-methylbutyll-L-histidinamide To a solution of the product from part (a) (194 mg., 0.236 mmole) dissolved in methanol (8.9 ml.) is added water (1.7 ml.), lN a~ueous hydrochloric acid (0.504 ml.) and 20% palladium hydroxide on carbon catalyst (150 mg.~. The mixture is stirred under an atmosphPre of hydrogen (balloon~ for 18 hours. It is then filtered and concentrated ln vacuo to give 153 mg. of crude product. Flash chromatography (LPS-l silica gel, 25 g.) eluting with chloroform:methanol:water:
acetic acid (90:20:2.5:1) followed by lyophilli-zation from 1% aqueous acetic acid of the pooledproduct containing fractions gives 124 mg. of N2-[N-C(1,1-dimethylethoxy)carbonyl]-L-phenyl-alanyl]-N-[(S)-l-[(S)-hydroxy(1~-imidazol-2-yl)-methyl]-3-methylbutyl]-L-histidineamide, acetic acid solvate; m.p. 105-110; [a]D ~ -30.1 1 3 1 07"2 HA375a (~ - 0.5, methanol). TLC Isilica gel; chloroform:
methanol:water:acetic acid, 90:20:2.5:1) Rf = 0.11.
Anal calc'd. fox C~gH41N7O5~ 1.3 C2H4O~ 2 C, 55.23; H, 7.45; N, 14.27 Found: C, 55.19; H, 7.20; N, 14.29.
Example 3 N2-[N-[(1,l-Dimethylethoxy)carbonyl]-L-phen~
alanvl]-N-[(S)~1-[(R)-hydroxy~lH-imidazol-2-yl)-methyll-3-methvlbutyl]-L-histidinamlde, acetic acid solvate a) N~-[N- L( 1,1-Dimethylethoxy)carbonyll-L-phenylalanyll~N-~(S)-l-[(R)-hydroxy[l [(phenyl-methoxy)-methyl]~lH-imidazol-2-yllmethyl]-3-methyl-butyll-3'- E (phenylmethoxy)methyl]-L-histidinamide) A solution of ~-[(S)-1-[[(1,l-dimethylethyl-ethoxy)carbonyl]amino]~3-methylbutyl]-1-[(phenyl-methoxy)methyl]-lH-imidazole-2-methanol,slow movin~
isomer, from Example l(b), (404 mg., 1 mmole) in ethyl acetate (15 ml.) is cooled in an ice water ba~h to 0 and saturated with gaseous hydrogen chloride. After remaining in the bath for lS
minutes, the bath is removed and the mixture is allowed to stand at ambient temperature for 45 minutes. Removal o the solvents ln vacuo gives ~5 396 mg. o~ solid a-[(S)-l-amino-3-methylbutyl]-1-[(phenylmethoxy)methyl]-lH-imidazole-2(R)-methanol, dihydrochloride.
To a mixture of this amine salt (0.81 mmole), N-~N~tl,l-dimethylethoxy)carbonyl]-L-phenylalanvl]-l'-[(phenylmethoxy?methyl]-L-histidine (423 mg., 0.81 mmole), and 1-hydroxybenzotriazole hydrate 1 3 1 07q2 HA375a (124 mg., 0.81 mmole~ in 6 ml. of dimethylformamide cooled in an ice-water bath under argon is added N-methylmorpholine (164 mg., 1.62 mmole) followed by dicyclohexylcarbodiimide (167 mg., 0.81 mmole).
The reaction mixture is refrigerated overnight, ~len diluted with ethyl acetate and filtered. The organic solution is rinsed with water, saturated sodium bicarbonate solution, and brine, dried over magnesium sulfate, and concentrated ln vacuo to give 566 mg. of crude product. Flash chromato-graphy (LPS-l silica gel, 60 g.) eluting with chloroform:methanol:ammonia (30:2:0.05) gives 270 mg. of N2-[N-[(1,1-dimethylethoxy)carbonyl]-L-phenylalanyl]-N-[(S)-l-[(~)-hydroxy~l-[(phenylmethoxy)-methyl]-lH-imidazol-2-yl]methyl]-3-methylbutyl]-3'-[(phenylmethoxy)methyl]-L-histidinamide;
m.p. 78-88; [~]D = -9.2 (c - 1, methanol). TLC (silica gel; chloroform:methanol:
ammonia, 30:2:0.5) Rf = 0.10.
Anal. calc'd. for C45H57N707:
C, 66.89; H, 7.11; N, 12.14 Found: C, 66.69; H, 7.18; N, 12.17.
b) N2-[N-[(1,1-Dimethylethoxy)carbonylL-L-phenyl-alanyl]-N-[(S)-1-[(R?~drox~y(1 -imidazol-2-yl)-methyll-3-methYlbutyl]-L-histidinamide To a olution of the product from part (a) (255 mg., 0.316 mmole) dissolved in methanol (11.7 ml.) is added water (2.2 ml.), lN agueous hydrochloric acid (0.663 ml.) and 20% palladium hydroxide on carbon catalyst (150 mg.). The mixture is stirred under an atmosphere of hydrogen HA375a (balloon) for 18 hours. It is then filtered and concentrated ln vacuo to give 208 mg. of crude product. Flash chromatography (LPS-1 silica gel, 35 g.) eluting with chloroform:methanol:water:
acetic acid (90:20:2.5:1)followed by lyophillization from 1% aqueous acetic acid of the pooled product containing fractions gives 144 mg. of N2-[N-[(l,1-dimethylethoxy)carbonyl]-L-phenylalanyl]-N-[(S)~1-[(R)-hydroxy(lH-imidazol-2-yl)methyl]-3~methylbutyl]-L-histidinamide, acetic acid solvate; _.
m.p. 194 - 196 (dec.); [~]D = 8.6 (c = 0.5, methanol). TLC (silica gel:chloroform:
methanol:water:acetic acid, 90:~0:2.5:1) Rf = 0.14.
Anal. calc~d~ for C29H41N7~5 1-3 C2H4~ 2 C, 55.22; H, 7.47; N, 14.36 Found: C, 55.15; H, 7.18; N, 14.31.
Example 4 N2-~N-L(l,1-Dimethylethoxy)carbonyl]-L-phenylalanyl]-N-r(S)-1-~S?-hydroxy(lH_imidazol-2_ l)methyl~-2-~henyl-ethYl~-L-histidlnamide, diacetate salt a) N-[(1,1-Dim_thylethoxy)carbonyll-L-Phenylalaninal To a solution of lithium borohydride (940 mg.
43.2 mmole) in dry tetrahydrofuran (90 ml.) cooled in an ice-bath under nitrogen is added a solution containing N-[(1,1-dimethylethoxy)carbonyl]-L
phenylalanine, N-hydroxysuccinimide ester (6.0 g., 16.6 mmole) [prepared according to the procedure of Anderson et al., JACS, Vol. 86, p.l839 (1964)] in tetrahydrofuran (60 ml.). The addition is carried out over a period of 5 minutes. After an HA375a additional 20 minutes at 0, the reaction mixture is poured into 1 1. of cold 10% potassium bisulfate.
The mixture is extracted with ethyl acetate (4 x 150 ml) and the combined organic extracts are rinsed with saturated sodium bicarbonate, water, and brine, dried over magnesium sulfate, and concentrated ln vacuo to give 3.8 g. of crude product. Flash chromatography (Merck 9385 silica gel, ~50 g.) eluting with chloroform:methanol (100:1, 50:1, and finally 25:1) gives 2.4 g. of purified product. Recrystallization from ether-hexane gives 1.9 g. of (S)-2-[[(1,1-dimethylethoxy)-carbonyl]amino]-2-phenylmethyl-1-ethanol; m.p.
95-96; [~]D = -27.5 (c = 1, methanol). TLC
(silica gel; chloroform:methanol:acetic acid, 25:1:1) Rf = 0.54.
Anal. calc'd. for C14H21NO3 C, 66.90; H, 8.42; Ni 5.57 Found: C, 67.02; H, 8.49; ~, 5.23.
~0 Pyridine sulfur trioxide complex (3.04 g., 19.1 mmole) is added to dry dimethylsulfoxide ~7 ml.) under argon and stirred at room temperature for 15 minutes. This mixture is then treated with a mixture of (S)-2-[[(1,1-dimethyl-~5 ethoxy)carbonyl]amino]-~-phenylmethyl-l-ethanol (1.~ g~, 4.77 mmole) and diisopropylethylamine 7 g., 19.1 mmole) in dry methylene chloride ~7 ml.), added rapidly along with the simultaneous application of an ice-water cooling bath. The ic~-bath is removed and after 10 minutes the reaction mixture is poured onto a mixture of 50 ml.

1 :~1 07~2 EA375a each of ice-water and ether. The aqueous portion is further extracted with ether (2 x 50 ml.) and the combined organic extracts are washed with 5%
potassium bisulfate, water, saturated sodium bicarbonate, water, and brine, dried over magnesium sulfate, and concentrated ln vacuo to give 949 mg. of N-[(1,1-dimethylethoxy) carbonyl]-L-phenylalaninal; m.p. 71-80;
[~]D = +37-7 (c = 1, methylene chloride).
TLC (silica gel; petroleum ether:acetone, 3:l) Rf = 0.45.
b) N - [ ( S ? -1- ~ ~ 51, 1-Dimethylethoxy ? carbonyl~amino]-2-phenylethyl-1-[(phenylmethoxy)methyll-lH-imidazole-2-methanol 2.5 M n-Butyllithium solution in hexane (2.8 ml., 6.99 mmole) is added to a solution of l-[(phenylmethoxy)methyl]-lH-imidazole (1.28 g., 6.82 mmole) in dry tetrahydrofuran (20 ml.) at -70 under argon. After 45 minutes at -70, a solution of N-[(1,1-dimethylethoxy)carbonyl]-L~
phenylalaninal (850 mg., 3.41 mmole) in tetra-hydrofuran (4 ml.) is added over a period of several minutes. After 2 hours at -70, the reaction is warmed to 0 and then guenched by the addition of saturated ammonium chloride (3 ml.).
The reaction mixture is then treated with ether (lO0 ml.) and water (1.5 ml.). The organic extract is rinsed with water (10 ml.) and brine, dried over magnesium sulfate, and concentrated ln 30 vacuo to give 2.04 g. of crude product. Flash ~~
chromatography (LPS-1 silica gel, 80 g.) eluting ~ 31 0~7n~
HA375a with petroleum ether:acetone (3:1) gives 234 mg.
oî ~-[~S)-l-[[(1,1-dimethylethoxy)carbonyl]amino]-2-phenylethyl-1-[(phenylmethoxy)methyl~-lH-imidazole-2-methanol (fast moving isomer), 152 mg. of a mixture fraction, and 294 mg. of a slow moving isomer. TLC (silica gel; petroleum ether:acetone, 2:1) Rf = 0.13, 0.08.
c) N~-[N- Ul,l-Dimethylethoxy)carbonYll-L-phenylalanyll-N- r ( s ~ hydroxy[l~ henyl-methoxy)methy,ll-lH-imidazol-2-yllmethyl]-2-phenylethyll-3'~phenylmethoxy)methy~l]-L- --histidinamide An approximately (1:1) mixture of the fast and slow moving isomer products from part (b) (512 mg., 1.17 mmole) is dissolved in ethyl acetate (20 ml.). The solution is cooled in an ice-water bath and saturated ~ith gaseous hydrogen chloride. The stoppered reaction is kept cold for o~e hour, and then concentrated ln vacuo to 492 mg. of liyht tan colored a-[(S)-l-amino-2-phenylethyl]-l-[(phenylmethoxy)methyl]-lH-imidazole-2-methanol, dihydrochloride.
This amine salt (1.17 mmole) is heated with l-hy~roxyben~otriazole hydrate (179 mg., ~5 1.17 mmole), N-[N-[(l,l-dimethylethoxy)carbonyl]-L-phenylalanyl]-l'-[(phenylmethoxy)methyl]-L-histidine (611 mg., 1.17 mmole), and dimethyl-~ormamide (9 ml.). The above mixture is cooled in an ice-bath under argon and treated with dicyclo-hexylcarbodiimide (237 mg., 1.17 mmole). The reaction mixture is refrigerated overnight, then HA375a diluted with ethyl acetate and filtered. The filtrate is diluted with ether and xinsed with water, saturated sodium bicarbonate, and brine, dried over magnesi~ sulfate, and concentrated ln S vacuo to give 882 mg. of crude product. Flash chromatography (LPS-1 silica gel, 120 g.) eluting with chloroform:methanol:ammonia (30:2:0.05) yields 160 mg. of N2~[N-[(l,l-dimethylethoxy)-carbonyl]-L-phenylalanyl]-N-[(S)-l-[hydroxy[1-~(phenylmethoxy)methyl]-lH-imidazol-2-yl]methyl]-~-phenylethyl]-3'-[(phenylmethoxy)methyl]-h-histidinamide (fast moving isomer), 239 mg. of the slow moving isomer, and about 60 mg. of a mixed fraction. TLC (silica gel; chloroform:methanol:
ammonia, 30:2:0.05) Rf = 0.28, 0.26. [~]D = -25.4 (c = 1, methanol) for the fast moving isomer and [~]D = -29.1 (c = 1, methanol) for the slow moving isomer.
d) N2-[N-r(l,l-Dimethylethoxy)carbonyll-L-phenyl-alanyll-N-[(S~ S)-hydroxy(lH-imidazo1-2-y~methyl]-2-phenylethy~l_L-histidinamine, diacetate salt A mixture of the fast moving isomer from part (c) (157 mg., 0.186 mmole), 20% palladium ~5 hydroxide on carbon catalyst (100 mg.), methanol (6.9 ml~), water (1.3 ml.), and lN aqueous hydro-chloric acid (0.391 ml.~ is stirred at room temperature under hydrogen (balloon) for 25 hours.
The reaction mixture is then filtered and 30 concentrated in vacuo to give 139 mg. of crude ~--product. Flash chromatography ILPS-silica gel, 13107~2 HA375a 21 g.) eluting with chloroform:methanol:water:
acetic acid (90:20:2.5:1) yields 107 mg. of N2-[N-[~l,l-dimethylethoxy)carbonyl]-L-phenylalanyl]-N-[(S)-l-[~S~hydroxy(lH-imidazol-2-yl)-methyl]-2-phenylethyl]-L-histidinamide, diacetate salt; m.p. 210-212 (d); [~]D = -21.4 (C = 0.5, methanol). TLC (silica gel; chloroform:
methanol:water:acetic acid, 90:20:2.5:1) Rf = 0.09.
Anal. calc~d. fox C32H39N7O5 2C~ 4 2 2 lOC, 57.06; H, 6.78; N, 12.94 Found: C, 57.01; H, 6.50; N, 12.94. ~-Example 5 N2- rN- r (l,l-Dimethylethoxy)carbonvl]~L-phenyl-alanyll-N-r(S)-l-[(R)-hydroxy(lH-imidazol 2-yl)-methyl~-2-phenylethyl]-L-histidinamide acetate salt ~1:1.5) A mixture of N2-~N-[(1,l-dimethylethoxy)-carbonyl]-L-phenylalanyl]-N-[(S)-l-[hydroxy[l-[(phenylmethoxy)methyl]-lH-imidazol-2-yl]methyl]-2-phenylethyl]-3'-~(phenylmethoxy)methyl]-L-histidinamide (slow moving isomer) (236 mg., 0.280 mmole) [prepared in Example 4(c)], 20%
palladium hydroxide on carbon catalyst (100 mg.), methanol (10.4 ml.), water (2.0 ml.), and lN aqueous hydrochloric acid (0.588 ml.) is stirred at room temperature under hydrogen (balloon) for 25 hours. It is then filtered and concentrated ln vacuo to give 193 mg. of crude product. Flash chromatography (LPS-l silica gel, 28 g.) eluting with chloroform:methanol:water:
acetic acid (90:20:2.5:1) yields 198 mg. of 1 3 1 07~2 HA375a product. Lyophilli~atio~ from 1% agueous acetic acid (polycarbonate filtered) yields 201 mg. of N -[N~ dimethylethoxy)carbonyl]-L-phenyl~
alanyl]-N-[(S)-l~[(R)-hydroxy~ imidazol-2-yl)methyl]-2-phenylethyl]-L-histidinamide, acetate salt (1:1.5); m.p. 1g6 - 217; [~]D = ~30 7 (c = 0.5, methanol). TLC (silica gel; chloroform:
methanol:water:acetic acid, 90:20:2.5:1) Rf = 0.09.
~nal. calc'd. for C32H39N7s 1-5 C2H42 2 C, 57.47; ~, 6.81; N, 13.41 Found: C, 57.47; H, 6.47; N, 13.40.
Example 6 "
N -[N-[(l,l-Dimethylethoxy)carbonYll~L-phenyl-alanyll-N-r(lS)-2-cyclohexy~ s-~-hy~ lH
lS imida~ol-2-vlmethylLethYl]-L-histidinamide, monoacetate salt a) (S~-2 LL~ _meth~ethoxy~arbonyl~amino 2-Phenylmethyl-1-ethanol To a solution containing N-~(l,l-dimethyl-ethoxy)carbonyl]-L-phenylalanine (lO g., 37.7 mmole) in dimethylformamide (40 ml.) is added solid sodium bicarbonate (4.75 g., 56.6 mmole~ and iodomethane (16 g., 113 mmole). The mixture is h~ated at ~0 under argon for 12 hours, ~he cooled ~S and the reaction mixture partitioned between water (150 ml.) and ether (250 ml.). The organic layer is rinsed with 2% aqueous sodium bicarbonat~
(2 x lO0 ml.), 2% aqueous sodium bisulfite (lO0 ml.),~water (2 x 100 ml.)~ and brine, dried over magnesium sulfate, and concentrated ln vacuo to give 10.5 g. of N-[(1,l-dimethylethoxy)carbonyl]-1 ~` 1 979 HA375a L-phenylalanine, methyl ester as an oil;
[~]D = +47-7 (c = 1, methylene chloride3 TLC (silica geli petroleum ether:acetone, 6:1) Rf = 0.41.
Anal. calc'd. for C15H21NO4:
C, 64.49; H, 7.58; N, 5.01 Found: C, 64.56; H, 7.60; N, 5.29.
To a solution containing N~[(l,1-dimethylethoxy)carbonyl]-L-phenylalanine, methyl ester (10 g., 35.8 mmole) dissolved in a mixture of tetrahydrofuran (190 ml.) and absolute ethanol (190 ml.) is added lithium chloride (6.09 g., 143.2 mmole). The resulting homogeneous solution is treated with sodium borohydride (5.42 g., 143.2 mmole) and the reaction is stirred at room temperature under argon for 24 hours. The reaction mixture is next filtered using ether (a'oout 700 ml.) to rinse the filter cake. The resulting filtrate is rinsed with water ( 3 x 200 ml.~ and brine (200 ml.), dried over magnesium sulfate, and concentrated 1n vacuo to give 9 g. of crude product. Recrystallization from ether/hexane gives 7.59 g. of (S)-2-[[(1,1-dimethylethoxy)carbonyl]amino]-2-phenylmethyl-1-ethanol; m.p. 94 - 96; [a]D ~ -27-2 (c = 1, methanol). TLC(silica gel; petroleum ether:acetone, 3:1) Rf = 0.39.
Anal. calc'd. for C14H21NO3:
C, 66.90; H, 8.42; N, 5.57 Found: C, 66.80; H, 8.57; N, 5.38.

- 1 3 ~ 07q2 HA375a b) [(S)-2-Cyclohexyl-1-(hydroxymethyl)ethyl carbamic acid, 1,1-dimethylethyl ester A solution of (S)-2-[[(1,1-dimethylethoxy)-carbonyl~amino]-2-phenylmethyl-1-ethanol (7 g., 27.8 mmole) in methanol (70 ml.) is hydrogenated at 55 psi on a Parr Shaker using 5% rhodium on alumina (500 mg.) as catalyst. After 17 hours, the reaction mixture is filtered and concentrated in vacuo to yield 7.36 g. of [(S)-2-cyclohexyl-1-(hydrox~ethyl)ethyl]carbamic acid, 1,1-dimethyl-ethyl ester as an oil; [~]D = -23.3 (c = 1, methylene chloride). TLC (silica gel; petroleum ether:acetone, 3:1) Rf = 0.5.
Anal. calc'd. for Ci4H27NO3:
C, 65.33; H, 10.57; N, 5.44 Found: C, 64.94; H, 10.55; N, 5.23.
c) (S)-(2-Cyclohexyl-1- I ormylethyl~carbamic acid, l,l-dimethyleth~l estex A solution of [(S)-2-cyclohexyl-1-(hydroxy-methyl)ethyl]carbamic acid, l,l-dimethylethyl ester ~4.6 g., 17.9 mmole) in methylene chloride (40 ml.) is added to a mixture of Dess-Martin periodinane reagent (8 g., 19 mmole) [prepared according to Dess et al., J. Org, Chem., Vol. 48, p. 4155 (1983)] and t-butanol (1.5 g., 19 mmole) in methylene chloride ~70 ml.) which had been stirred at room temperature before the addition.
A slight exotherm (to 32) results. After 30 minutes, the reaction mixture is quenched in ether (800 ml.), resulting in the separation of a white solid. A mixture of sodium thiosulfate pentahydrate 7 ~ ,~
HA375a (31.3 g., 126 mmole) in saturated sodium bicar-bonate solution (200 ml.) is added, with stirring.
The resulting two-phase mixture is separated and the oxganic phase is washed with water, saturated sodium bicarbonate (2 x 100 ml.), water, and brine, dried over magnesium sulfate, and concentrated ln vacuo to give 3.8 g. of (S)-(2-cyclohexyl-l-formylethyl)carbamic acid, 1,1 ~imethylethyl ester as a colorless oil.
1~ d) ~(lS)-l-(Cyclohexylmethyl)-2-hydroxY-2-~1-[(phenvlmethoxy)methylJ-lH-imidazol-2-yl]-ethyl~carbamic acid, l,1-dimethyleth~l ester 2.5 M n-Butyllithium solution in hexane (12 ml./ 31 mmole~ is added to a solution of l-[(phenylmethoxy)methyl]-lH-imidazole ~5.3 g., 28 mmole) in tetrahydrofuran (90 ml.) at -70 under argon. After stirring for 15 minutes, (S)-(2-cyclohexyl-1-formylethyl)carbamic acid, l,l-dimethylethyl ester (3.6 g., 14 mmole) in 0 tetrahydrofuran (36 ml.) is added dropwise over a period of 5 minutes at a reaction temperature of -65 to -70. After 2 hours at ~70, the bath is warmed to 0 and saturated ammonium chloride (25 ml.) is added followed by ether (300 ml.) and water (25 ml.). The organic phase is washed with water (2 x 50 m.) and brine, dried over magnesium sulfate, and concentrated ln vacuo. The resulting crude product (8.4 g.) is flash chromatographed ~LPS-l silica gel) eluting with acetone:petroleum ether ~1:4) to give 580 mg. of [(lS)-1-(cyclo- ~
hexylmethyl)-2-hydroxy-2-[1 [(phenylmethoxy)methyl]-~ 3 ~ O~q2 HA375a lH-imidazol-2-yl]ethyl]carbamic acid, 1,1-dimethylethyl ester (fast moving isomer), 370 mg.
of a mixed fraction, and 2 g. of a slow moving isomer. TLC (silica gel; acetone:petroleum ether 1:4) Rf = 0.15, 0.10.
Fast moving isomer; [~]D = -21.5 (13 mg./ml., methanol).
Anal.calc'd. for C25H37N3O4 2 C, 66.61; H, 8.45; N, 9.32 Found: C, 66.55; H, 8.39; N, 9.00.
Slower moving isomer; [~]D = +9.1 -' (14 mg./ml., methanol).
Anal- calc'd. for C25H37N304 022 H2O
C, 67.08; H, 8.43; N, 9.39 Found: C, 67.08; ~, 8.35; N, 9.01.
e~ N2-~N-[(l~l-Dimethylethoxy)carbonyl]-L-henylalanyll-N-~(S~-l-r(S)-hydroxy11-[(phenyl-methoxy)methyl~-lH-imidazol-2-~l]methYl]-?-cyclohexylethyll~3'-[(phenylmethox~methyl~-L-histidinamide A solution of the fast moving isomer from part (d) (430 mg., 0.97 mmole) in ethyl acetate (8 ml.) is cooled in an ice-water bath and saturated with gaseous hydrogen chloride. After remaining in the cold for 15 minutes, the mixture is kept in a stoppered flask at ambient temperature for 45 minutes. The mixture is concentrated ln vacuo to give 384 mg. of the amine dihydrochloride salt.
This amine salt (306 mg., 0.8 mmole) is dissolved in dry dimethyformamide (6 ml.) and 13tO7q2 HA375a N-[N-[(l,l-dimethylethoxy)carbonyl]-L-phenyl-alanyl]-1'-[(phenylmethoxy)methyl~-L-histidine (442 mg., 0.8 mmole) is added followed by l-hydroxybenzotria201e hydrate (122 mg., 0.8 mmole) and N-methylmorpholine (160 mg., 1.6 mmole). After cooling in an ice-water bath, dicyclohexylcarbodiimide is added. The mixture is ~tirred in the cold for 15 minutes and then refrigerated overnight in a stoppered flask. The solids that separate ~rom the solution are recovered by filtration after the mixture is diluted to a volume of 45 ml. with ether. The filtrate is washed with water, sa~ura~ed sodium bicarbonate solution, and brine, dried over magnesium sulfate, and concentrated in vacuo to give a viscous oil residue (587 mg.). This residue is preabsorbed on silica gel (Baker's, 3 g.~ and flash chromatographed (LPS-1 silica gel, 90 g.) eluting with chloroform:methanol:concentrated ~0 ammonium hydroxide ~30:2:0.05) to give reco~ery of the product in two fractions (333 mg. of impure and 70 mg. of pure product). The impure fraction is rechromatographed as above to give a recovery o~
206 mg. This is combined with the 70 mg. fraction ~S and chromatographed again according to the above procedure to give 255 mg. of N2-[N-[(l,1-dimethyl-e~hoxy)carbonyl]-L-phenylalanyl]-N-[(lS)-1-[~S)-hydroxy-[l-[(phenylmethoxy)methyl]-lH-imidazol-2-yl]methyl]-2-cyclohexylethyl]-3'-[~phenylmethoxy)-mathyl~-L-histidinamide as a glassy solid;
m.p. 74-77 ~gradual melt); [a]D = -24.3 1 3 ~ 079~

~3753 . SD~ - .

~c = 1, methanol3. TLC (silica g~l: chloroo~s math3nol:conc. ammo~iu~ hydroxid~, 30:2 0.0 R~ = û.17.
Anal . calc ' d- for C4~3E3t61N7~7 1E~2 C, 66,57; ~, 7.33; ~ 11.32 Found: C, 66.55; E~, 7.26; N, 11.37.
f) ~__ A mixt~re~ of th~ produc~ ~o~
part ~) (24~ mg., 0.~8 m~ola) ~ 20% ~alladl~u~
h~dro~ida oQ car~o~ ~ataly~ 185 ~g.) in m~ o~
~i ml.~ ~lu5 wat~r (0.~1 ~O) ~nd l~t h~lkos:l~lor~c acid (0.62 ml., 0.62 ~oles) i4 8tl~0~ at roo~a tllperatur~ in a~ at~o~ o~ hydrog~~ lloo~
for 60 hour~ addition~l 100 n~. s:~ ca~ly~t and 4 ml. o~ m*thanol ar~ ad~3d a~
stir~ed in a~ atmosph~ of hy~go~ ~or 20 hour~. After filterin51 ~ough~ite~, tl~
filt~at2 i~ eonc~era~d i_ ~ to qiv~$ 200 mg, o~ ~ gl~s~y solid seEI~du~. Thi~ ros~ldu~ i~ th~
fla~h c~ro~atoslraph~a ~I,PS~ ca ~ 0 sr~) ~lutiny wit~ c3~10ro~on~:mothanol :wat~s a~otlc: ~id ~9Os20:2.Ssl) to gi~ 120 Dl~. o~ h ~ r no~,id 25 ~ ak i~ lyol?hilliz~d froDI~ 3X alC~ cia to givo _120 mg. o~ an amorphous whits Jol~d. Thi~ ~olid i~ ~o~hro~atograph~c~ (IPS-l ~ilic~ gol, 16 ~. ) eluting with chloroform:m~ ol:w~t~r:ac~
acid ~90:20:205:1) to give~ 69.4 m~. of N2 tN~
30 dimcthyl~l:ho~y~carbo~yl~-L-phsnylalanyl]-N-t~15) 2-* Trademark ~ .

~ ' ,.

1 3 1 079~

HA375a cyclohexyl-1-[(S)-hydroxy-lH-imidazol-2-ylmethyl]-ethyl] L-histidinamide, monoacetate salt; m.p. 162 (gradual melt, shrinks at 110);
[~]D = -35.6 (5 mg./ml., methanol). TLC (silica gel; n-butanol:pyridine.acetic acid:water, 4:1:1:1) Rf = 0.61.
Anal. calc'd- for c32H45N7o5 1 C2 4 2 C, 57.07; H, 7.65; N, 13.70 Found: C, 57.05; H, 7.40; N, 13.72.
10Example 7 N2 [N-~(l,l-Dimethylethoxx~ y~
alanyll-N-~lS)-2-cy~clohexyl-l-[(R)-hydroxy-lH
imidazol-2-ylmethyllLethyll-L-histidinamide~
monoacetate salt a) N2-rN-r(l,l-DimethYlethoxy)carbonyll-L-phenylalanyl]-N-[(lS)-l- L t R)-hydroxv~ phenYl-methoxv)methyl~-lH-imidazol-2-yl]meth~yl]-2-cyclohexylethyll-3~ henylmetho-xy)me-thyl]-L
histidinamide A solution of [(lS)-1-(cyclohexylmethyl)-2-hydroxy-2-[1-[(phenylmethoxy)methyl]-lH-imidazol-2-yl]ethyl]carbamic acid, 1,1-dimethylethyl ester (slow moving isomer) (467 mg., 1,05 mmole) [prepared in Example 6 (d)]
in ethyl acetate (25 ml.) is cooled in an ice-water bath under argon and saturated with gaseous hydrogen chloride. The stoppered reaction is kept cold for one hour and then concentrated ln vacuo to give 467 mg. of the amine dihydrochloride salt.
This amine salt (374 mg., 0.84 mmole) is ~~
dissolved in dimethylformamide (6 ml.) along with ~310:7~
HA375a N-[N-[(l,l-dimethylethoxy)carbonyl]-L-phenyl alanyl]-l'-[(phenylmethoxy)methyl]-L-histidine (439 mg., 0.84 mmole) and l-hydroxybenzotriazole hydrate (128 mg., 0.84 mmole). The mixture is cooled under argon in an ice-water bath and treated with N-methylmorpholine (170 mg., 1.68 mmole) followed by dicyclohexylcarbodiimide (173 mg., 0.84 mmole). The stoppered reactlon mixture is refrigerated o~ernight, then filtered and extracted with ethyl acetate. The organic ~.
solution is rinsed with water, saturated sodium bicarbonate, water, and brine, dried over maynesium sulfate, and concentrated ln vacuo to 730 mg. of crude product. Flash chromatography (LPS-l silica gel, 70 g.) eluting with chloroform:methanol:
concentrated ammonia (30:2:0.05) gives 271 mg.
of N2-[N-[(1,1-dimethylethoxy)carbonyl]-L-phen~l-alanyl]-N-[(lS)-l-[(R)-hydroxy[l-[(phenylmethoxy)-methyl]-lH-imida~ol-2-yl]methyl]-2-cyclohexyl-ethyl]-3'-[(phenylmethoxy)methyl]-L-histidinamide;
[~]D = -8.7 ~c = 1, methanol).
TLC ~silica gel; chloroform:methanol:conc. ammonia, 30:2:0.05) R~ = 0.14.
Anal. calc'd. for C48H61N7O7:
~5 ~, 67.98; H, 7.25; N, 11.56 Found C, 67.80; H, 7.27; N, 11.44.
b) N -[N-~(l,l-Dimethylethoxy ? carbonyl]-L-phenyl~
alanyl~-N-[(lS ? -2-cyclohexyl-1-[(R~-hydroxy-lH-imidazol-2-ylmethylle-hyl]-L-histidinamide 3~ monoacetate salt ~`
.
The product from part (a) (265 mg., 0.312 mmole) is dissolved in methanol (11.6 ml.) 1 3 1 07q2 _57_ HA375a to which is added water (2.2 ml.) followed by lN aqueous hydrochloric acid ~0.66 ml.) and 20%
palladium hydroxide on carbon catalyst (100 mg.).
The mixture is stirred under a hydrogen atmosphere (balloon) for 17 hours, then filtered and concentrated in vacuo to give 250 mg. of crude product. Flash chromatography (LPS-l silica gel, 35 g.) eluting with chloxoform:methanol:water (tap distilled):acetic acid (90:20:2.5:1) followed by lyophillization of the product containiny fractions gives 186 mg. of material. Rechroma- --tography using double distilled water (LPS-l silica gel, 20 g.) eluting with the 90:20:2.5:1 solvent system gives 95 mg. of N2-[N-[(l,1-dimethylethoxy)carbonyl]-L-phenylalanyl]-N-[(lS)-2-cyclohexyl-1-[(R)-hydroxy-lH-imidazol-2-ylmethyl]ethyl]-L-histidinamide, monoacetate sal~; m.p. 193 - 197 (d 200);
[~]D = -19 (c = 0.5, methanol). TLC (silica gel; chloroform:methanol:water:acetic acid, 90:20:2.5 1) Rf = 0.07.
Anal- calc'd- for c3~H45N7O5 0-8 C2 4 2 C, 59.26; H, 7.55; N, 14.40 Found: C, 59.24; H, 7.49; N, 14.37.
25Example 8 N-[(lS?-2-C clohex~l-l-[(R)-hydroxY(lH-imidazol-2-yl)~
methyl]ethy~ N2~[N~ l-dimethylethoxy)carbonyll-L
phenylalanyll-L-leucinamide~ acetate salt a) N-[N-r(l,l-Dimethvlethoxy~ rbonyl~-L-30 ~ phenylalanyl]-L-leucine~ methyl ester A solution of diisopropylethylamine (8.7 ml., 50 mmole) in tetrahydrofuran (50 ml.) is added dropwise to a mixture of N-[(l,1-dimethylethoxy)-carbonyl~-L-phenylalanine (13.265 g., 50 mmole), L-lPucine, methyl ester (9.085 g., 50 mmole) and 1 3 1 07~2 HA375a l-hydroxybenzo~riazole hydxate (7.65 g., 50 mmole) in tetrahydrofuran (lO0 ml.) at 0. This is followed by the addition of dicyclohexylcarbo-diimide (10.315 g., 50 mmole). The reaction is stirred at 0 for 2 hours and left stirring overnight at room temperature. The next day, the precipitated dicyclohexyl urea is filtered off, the solvents are stripped down and the residue is diluted with ethyl acetate (200 ml.). The organic solution is washed sequentially with saturated aqueous sodium bicarbonate solution (2 x 100 ml.) and saturated aqueous sodium chloride (2 x lO0 ml.), dried over sodium sulfate, filtered, and concentrated to give crude product.
~5 Crystallization from ethyl ether gives 7.05 g. of pure product. Concentration of the mother liquor solutions gives 4.57 g. of crystalline product. An additional I.35 g. of product is obtained by chromatographic purification of the crude product obtained from the left-over mother liquors (40 g.
silica gel, eIuting with 4:1 hexane:ethyl acetate).
Thus, a total of 12.96 g. of N- [N- [(1,1-dimethyl-ethoxy)carbonyl]-L-phenylalanyl]-L~leucine, methyl ester is obtained; m.p. 104 - 105; [~]D = -17.5 ~5 (c = 1.2, methanol). TLC (silica gel; hexane:
ethyl acetate, 1:1) Rf - 0.57.
Anal. calc'd. for C21H32N2O5:
C, 64.30; H, 8.15; N, 7.14 Found: C, 64.12; H, 8.16; N, 7.02.
b) N-~LlS)-2-Cyclohexyl-l-[(R)-hydroxy[l-[(pheny methoxy)methyl]-lH-imidazo N2-[N-~(l,l-dimethylethoxy)carbonyl]-L-~henyl-alanyl]-L-leucinamide The methyl ester product from part (a) (1.176 g., 3 mmole) in tetrahydrofuran (12 ml.) HA375a is treated with aqueous lN sodium hydroxide (3.3 ml., 3.3 mmole). After two hours at room temperature, the mixture is refluxed for two hours The solvents are stripped down and the residue is taken up in water, acidified, and extrac~ed with ethyl aceiate. The organic layer is dried over sodium sulfate and concentrated to give a residue which is resubjected to hydrolysis with aqueous lN sodium hydroxida (1.5 ml., 1.5 mmole) in methanol (12 ml.). After three hours at room temperature, the solvents are stripped down. The residue is taken up in water ~100 ml.), extracted with ethyl acetate t2 x ~5 ml.), and the organic layer is discarded.
The aqueous layer is carefully acidified (pH 3.9), extracted with ethyl acetate (3 x 25 ml.), dried over sodium sulfate and concentrated to give ~53 mg. of N-[N-(1,1-dimethylethoxy)carbonyl]-L-phenylalanyl]-L-leucine.
~0 To a mixture of this acid (753 mg., 2.0 mmole), [(lS)-1-(cyclohexylmethyl)-2-hydroxy-2-[l-[~phenylmethoxy)methyl]-lH-imidazol-2-yl]ethyl]-carbamic acid, l,l-dimethylethyl ester, slow isomer ~rom Example 6(d) (887 mg., 2.0 mmole), and ~S l-hydroxyben~otriazole hydrate (306 mg., 2.0 mmole) in ~etrahydrofuran (8 ml.) at 0 is added diiso-propylethylamine (731 ~1., 4.2 mmole) followed by dicyclohexylcarbodiimide (416 mg., 2.0 mmole).
T~e reaction mixture is stirred at 0 for about 2 hours and then kept overnight in the cold room (about 5). The next day, the precipitated dicyclo~
hexyl urea is filtered off and the residue is redissolved in ethyl acetate, washed with saturated aqueous sodium bicarbonate solution (2 x 30 ml.) and saturated aqueous sodium chloride HA375a (2 x 30 ml.), dried over sodium sulfate, filtered, and c~ncentrat~d to give 1.339 g. of a crude residue. Repeated flash chromatographies (silica gel, eluting with 9:1:0.1 and 15:1:0.05 chloroform:methanol:acetic acid) gives 1.021 g. of N-[(lS)-2-cyclohexyl 1-[(R)-hydroxy[l-[(phenylmethoxy)-methyl]-lH-imidazol-2-yl~methyl]ethyl]-N2-[N-[(1,1-dimethylethoxy)carbonyl]-L-phenylalanyl]-L-leucinamide.
TLC (silica gel; chloroform:methanol:acetic acid, 9:1:0.1) Rf = 0.5.
c) N-~(lS) c-~-lo>e~y~ l IlR)-hydroxy~lH-imidazol-2-yl)methyl]ethyl]-N2-[N-~(l,l-dimethylet oxy)car-bonyl]-L-phenvlalanyll-L-leucinamide, acetate salt 20% Palladium hydroxide on carbon catalyst (100 mg.) is weighed in a side arm flask and flushed with hydrogen using a balloon. Ths L=leucinamide product from part (b) (295 mg., 0.42 mmole) is added via a syringe as a solution in methanol (5 ml.). This is followed by sequential addition of mathanol (8.5 ml.~, water (3.0 ml.), and aqueous 10% hydrochloric acid (0.307 ml.). The flask is then carefully flushed several times with hydrogen and stirred overnight at room temperature.
The contents of the flask are filtered and the filtrate is concentrated. The resulting residue is flash chromatographed (30 g. of silica gel;
elutins with 90:10:1:0.1 chloroform:methanol:
water:acetic acid) to give 155.3 mg. of N-[(lS)-2-cyclohexyl-1-[(R)-hydroxy(lH-imidazol-2-yl)methyl]-ethyl]-N2-[N-[(1,1-dimethyle~hoxy)carbonyl]-L-phenylalanyl~-L-leucinamide, acetate salt; m.p. 101-109 (gradual melting); [a]D = -25.7 (c = 1.06, methanol). TLC (silica gel; chloroform:

~ 3 t 07q2 HA375a methanol:water:acetic acid, 90:20:2.5:0.1) Rf = 0.21.
Anal- calc'd- for C32H~9N55 0 9 C2 4 2 2 C, 6~.33; H, 8.37; N, 10.75 Found: C, 62.38; H, 8.17; N, 10.75.

Example 9 N-[(lS)-1-(Cyclohexylmethyl)-2(R)~hydroxy~2-(lH-imidazol-2-yl)ethyl]-N2-[N-[(l,1-dimethylethoxy)-carbonyl~-L-phenylalanyl]glycinamide, acetate salt a) N-[N-[(l,1-Dimethylethoxy)carbonyl]-L-~henyl-alanyl~glycine~ ethyl ester To a mixture of N-[(1,1-dimethylethoxy)- -carhonyl]-L-phenylalanine (13.265 g., 50 mmole), glycine, ethyl ester, monohydrochloride (6.98 g., 50 mmole), and l-hydroxybenzotriazole hydrate ~7.65 g., 50 mmole) in tetrahydrofuran (100 ml.) at 0 is added dicyclohexylcarbodiimide (10.315 g., 50 mmole~ as a solution in tetrahydrofuran (25 ml.).
This is followed by the addition of a solution of diisopropylethylamine (8.7 ml., 50 ~mole) in tetra-hydrofuran (25 ml.). The rea`ction is stirred at 0 for 2 hours and then stirred ovexnight at room temperature. The next day, the precipitated dicyclohexyl urea is filtered off and the solvents ~5 stripped down. The residue is diluted with ethyl acetate (200 ml.) and the resulting organic 301ution is washed sequentially with saturated a~ueous sodium bica.rbonate solution (2 x 100 ml.) and saturated aqueous sodium chloride (100 ml.), dried over sodium sulfate, filtered, and concen-trated. The resulting crude material is crystall-ized from ethyl ether to give 9.45 g. of pure product. Concentration of the mother li~uor 1 3 1 07"2 HA375a solution to half its original volume gives an additional 3.7 g. of N-[N-[(1,1-dimethylethoXy)-carbonyl]-L-phenylalanyl]glycine, ethyl ester;
m.p. 90~9~; [~]D = -9.1 (c = 1.41, methanol).
Anal. calc'd. for C18H26N2O5:
c, 61.73; ~, 7.42; N, 7.99 Found: C, 61.64; H, 7.46; N, 8.07.
b) N-[(lS)-l-(Cyclohexylmethyl)-2(R)-hydroxy-~-[1-[(Phenylmethoxy)methyl]-lH-imidazol-2 ethyll-N2-[N-[(l,l-dimeth~lethoxy~carbonyl]-L~
~henylalanyl]glycinamide A solution of the ethyl ester product from part (a) (1.05 g., 3.0 mmole) in tetrahydrofuran (12 ml.) is`treated with agueous lN sodium hydroxide (3.1 ml., 3.1 mmole). ~ydrolysis is found to be complete after 2 hours at room tempera-ture. The solvents are stripped down and the residue is taken up in saturated agueous sodium bicarbonate solution (15 ml.) and extracted with ether (30 ml.). Some sodium salt precipitates at this stage and is filtered and separated from the biphasic layer. The ethereal layer is discarded.
The aqueous layer is carefully acidified (pH 5.0 and finally 2.5) and reextracted with ethyl acetate (3 x 20 ml.). The precipitated sodium salt is taken up in water, the aqueous solution is acidified (pH 2.5), and extracted with ethyl acetate (3 x 20 ml.). The combined organic extract is dried over sodium sulfate, filtered, and con-centrated to give 599 mg. of N-[(l,1-dimethyl-ethoxy)carbonyl]-L-phenylalanine]glycine.
To a mixture of the above acid (plus an added amount from a small scale run) (644 mg., 2.0 mmole) and [(lS)-1-(cyclohexylmethyl)-2-hydroxy-2-1 31 07q2 HA375a [l-[(phen~lmethoxy)methyl]-lH-imidazol-2-ylJ-ethyl]carbamic acid, l,l-dimethylethyl ester, slow isomer from Example 6(d) (887 mg., 2.0 mmole) in dimethylformamide ~8 ml.) at 0 is added l-hydroxy ben~otriazole hydrate (306 mg., 2.0 mmole), diiso-propylethylamine (731 ~1., 4.~ mmole), and finally dicyclohexylcarbodiimide (420 mg., 2.0 mmole). The reaction mixture is stirred for 2 hours at 0 and then overnight at room temperature. The next day, the precipitated dicyclohexyl urea is filtered off, and the filtrate is concentrated. The ~
residue is taken up in ethyl acetate (75 ml.) and washed sequentially with water (2 x 25 ml.), saturated aqueous sodium bicarbonate (2 x 25 ml.), and saturated aqueous sodium chloride (25 ml.), dried over sodium sulfate, a~d concentrated to give 838 mg. of crude product. Repeated chroma-tographic purifications (silica gel, eluting with 19:1 chloroform:methanol) gives 113 mg. of N-[(lS)-1-(cyclohexylmethyl)-2(R)-hydroxy-2-[1-[(phenylmethoxy)-methyl]-lH-imidazol-2-yl]ethyl]-N2-[N-[(1,l-dimethyl-ethoxy)carbonyl]-L-phenylalanyl]glycinamide.
TLC (silica gel; chloroform:methanol:
acetic acid, 9:1:1) Rf - 0.3.
Anal. calc'd. for C36H49N5O6:
C, 66.81; H, 7.62; N, 10.81 Found: C, 64.49; H, 7.41; N, 10.02.
c) N-~(lS)-l-(Cyclohex~lmethyl)-2(~)-hydroxy-2-tlH-imidazol-2-yl)ethyll-N2-[N-[(l,l-dimethyl-ethoxy)carbonyll-L-phenylalanyl]glycinamide~
acetate salt 20% Palladium hydroxide on carbon catalyst (100 mg.) is weighed in a side arm flask and flushed with hydrogen using a balloon. The glycinamide product ~ 1 0 7 q 2 HA375a from part (b) (118 mg., 0.2 mmole) is added as a 2 ml. methanol solution. This is followed by the sequential addition of methanol (6 ml.), water (2 ml.) and 10% aqueous hydrochloric acid (145 ~1.).
~he reaction mixture is stirred at room temperature for several days. The catalyst is removed by filtration and ~he filtrate is concentrated to give a crude product which after repeated ~hromatographic separations (silica gel, eluting with chloroform:methanol:water:acetic acid, 90:20:2.5:1) yields 9.5 mg. of N-[(lS)-l-(cyclo-hexylmethyl)-2(R)-hydroxy-2-(~H-imidaæol-2-yl)ethyl]-N2-[N-[(l,l-dimethylethoxy)carbonyl]-L-phenyl-alanine~glycinamide, acetate salt;
m.p. 99 - 100; [~D = ~7-0 (c = 0.1, methanol).
TLC (silica geli chloroform:methanol:water:acetic acid, 90:20:2.5:1) Rf = 0.27.
Anal calc~d for C28H41N55 0 7 C2 4 2 2 C, 60.16; H, 7.86; N, 11.92 ~0 Found: C, 60.06; ~, 7.64; N, 11.93.
Exam~le 10 N2-rN-r(l,l-Dimethylethoxy~L~arbonvl]-L-~henylalanylL-N-[(S)-1-r(R~-hvdroxy(2-thia~olyl)methyl]-3-methyl butvl]-L-histidinamide, acetate salt ~5 a) (2S~-2-rr(1,l-Dimethylethox~ bonyl]aminol--methyl-1-(2-thiazolyl2-1-pentanol 2.5N n-Butyllithium in hexane (8 ml.) is added to a solution containing thiazole (1.7 g., ~0 mmole) in dry tetrahydrofuran (60 ml.) cooled to -70 under argon. After stirring a short time, the solid material begins to come out of 1 3 i 0792 HA375a solution. The reaction remains heterogeneous after 35 minutes at -40 to -35. After cooling the reaction to -50, a solution containing N-[(1,1-dimethylethoxy)carbonyl]-L-leucinal (2.2 g., 10 mmole) in tetrahydrofuran (5 ml.) is added.
After 90 minutes the reaction is warmed to -10 and quenched with saturated ammonium chloride (10 ml.).
The reaction mixture is extracted into ether and the organic extract is rinsed with water and brine, dried (MgSO4) and concentrated in va~uo to give 1.8 g. of amide product. Two flash chromato-graphies on first 100 g. and then 150 g. o silica gel (LPS-l) eluting with petroleum ether:acetone (20:1) fails to remove a minor impurity from the co-eluting desired diastereomeric product mixture. A total of 1.2 g. of (2S)-2-[[1,1-dimethylethoxy)carbonyl]amino]-4-methyl-1-(2-thiazolyl)-l-pentanol is obtained. TLC (silica gel; petroleum ether:acetone, 4:1) Rf = 0.37.
[~]D = -38.6 (c = 1, chloroform).
b) [(l,l-Dimethylethoxy)carbonYl~-N -(2,4-dinitro~henyl)-N-[(S~ (R~ydroxy(2-thiaæol~l)-methyl]-3-methvlbutyll-L-histidinamide A solution of the product from part (a) (1.5 g., 5 mmole) dissolved in ethyl acetate (40 ml.) is cooled in an ice water bath under argon. After saturating with dry HCl gas, the reaction is kept cold or 30 minutes. After removal of the ethyl acetate ln vacuo, trituration of the residue gives 1.45 g. of crude bis hydrochloride salt.

1 3 1 079~

HA375a -6~-This bis hydrochloride salt (860 mg., 3.15 mmole), N-[(l,l-dimethylethoxy)carbonyl]-Nl-(2,4-dinitrophenyl)-L-histidine (1.38 g., 3.15 mmole), and l-hydroxybenzotria7O1e hydrate (482 mg., 3.15 mmole) are stirred in tetrahydro-furan ~30 ml.) under argon; cooled in an ice water bath, and treated with N-methylmorpholine (637 mg., 6.3 mmole) followed by dicyclohexylcarbodiimide (650 mg., 3.15 mmole). The reaction is stirred in the ice bath for a~ hour, refrigerated overnight, and then filtered. The filtrate is diluted with ethyl acetate. The organic extract is rinsed with water, saturated sodium bicarbonate, and brine, dried (MgSO4), and concentrated ln vacuo to give 1.82 g. of crude product. Flash chromatography on silica gel (170., LPS-l) eluting with chloroform:
methanol (20:1) yields 0.46 of fast moving isomex (S), 0.57 g. of a mixture of isomers, and 0.27 g.
of slow moving isomer (R). Rechromatographing the mixture ~raction gives a total of 0.54 g. of (S) isomer and 0.80 g. of r(l,l-dimethylethoxy)-carbonyl]-Nl-(2,4~dinitrophenyl)-N-[(S)-1-[(R)-hydroxy(2-thiazolyl)methyl]-3-methylbutyl]-L-histidinamide; m.p. 82 - 107. TLC (silica gel;
chloroform:methanol 20:1) Rf = 0.15. [~]D =
+12.7 (c = 1, methylene chloride).
c) N -rN-[(1,1-Dimethylethoxy2carb~ -L-p_enyl-alanyll-N-[(S)-l-[(R)-hydroxY(2-thiazolVl)methYll-3-methylbutyll-Nl-(2,4-dinitrophenyl)-L-histidlnamide A solution containing the (R) hydroxy isomer product from part (b) (510 mg., 0.84 mmole) in 1 3 1 ~7~

HA375a -67~

ethyl acetate (25 ml.) is cooled in an ice water bath and saturated with dry HCl. After stirring for 60 minutes, the solvent is removed in vacuo to yield 436 mg. of Nl-(2,4-dinitrophenyl)-N-~(S)-l-S [(R)-hydroxy(2-thiazolyl)me~hyl]-3-methylbutyl]-L-histidinamide, hydrochloride salt.
This crude hydrochloride salt (436 mg., O.46 mmole), N-[(1,l-dimethoxyethoxy)carbonyl]-L-phenylalanine (122 mg., O.46 mmole), and 1-hydxo~y-benzotria~ole hydrate (70 mg., 0.46 mmole) are _-stirred in tetrahydrofuran (10 ml.) under argon, cooled to 0 and treated with N~methylmorpholine (118 mg., l.lS mmole) followed by dicyclohexyl-carbodiimide (95 mg., 0.46 mmole). The reaction is allowed to warm to room temperatuxe overnight, then filtered and diluted with ethyl acetate and ether. The organic extract is rinsed with water, saturated sodium bicarbonate, and brine, dried (MgSO4), and concentrated 1n vacuo to give 360 mg.
~0 of crude product. Flash chromatography on silica gel (40 g., LPS-l) eluting with chloroform:methanol (20:1) yields 250 mg. of N2-[N-[(1,1-dimethylethoxy)-carbonyl]-L-phenylalanyl]-N-[(S)-l-[(R)-hydroxy-(2-thia~olyl)methyl]-3-methylbutyl]-Nl-(2,4-~5 dinitrophenyl)-L-histidinamide, m.p. 100 ~ 115;
[~]D = +25.5 (c = 1, chloroform). TLC(silica gel;
chloroform:methanol, 10:1) Rf = 0~32.
~n~l~ calc d. for C35H42N8OgS 1.5 H2O:
C, 54.04; H, 5.R3; N, 14.41; S, 4.12 Found: C, 53.99; H, 5.59, N, 13.97; S, 4.18.

1 3 ~ 0792 HA375a d) N2-[N-[(l,l-Dimethylethoxv)carbonyl]-L-phenyl-alanyl]~N-L~ 1-L(R)-hydroxy(2-thiazoly~)methyl]-3-methylbutyll-L-histidinamide, acetate salt Mercaptoacetic acid (1.15 g., 12.5 mmole) i5 added to a solution of the product from part (c) (243 mg., 0.324 mmole) in dimethylformamide (3 ml.
under argon. After 2 hours at room temperature, the reaction is extracted into a mixture of ethyl acetate (40 ml.) and ether (20 ml.), rinsed with seven 10 ml. portions of 10% a~ueous sodium carbonate, three 10 ml. portions of water, and brina. The organic extract is dried (MgSO4) and concentrated ln vacuo to 0.26g. of crude product. Flash chromatography on silica gel (25 g., LPS - 1) eluting with chloroform:
methanol:water:acetic acid (90:15:1:0.5) gives 112 mg. of product. This material is dissolved in 0.5% aqueous acetic acid ~20 ml.), millipore filtered, and lyophillized to give 99 mg. of N2-[N-[(l,l-dimethylethoxy)carbonyl]-L-phenylalanyl]-N-[(S)-l-[(R)-hydroxy(2-thiazolyl)methyl]-3-methyl-butyl]-L-histidinamide, acetate salt; m.p. 89-115;
[~]D = -30.2 (c = 0.5, methanol). TLC (silica gel; chloroform:methanol:water:acetic acid, 90:15:1:0.5) Rf = 0.21.
Anal. calc'd for C29H40N6O5S 0.5 C2 4 2 2 C, 56.94; H, 7.01; N, 13.28; S, 5.07 Found: C, 57.00; H, 6.63; N, 13.28; S, 4.78.

I ~` 1 07"~

HA375a Example 11 N-[(S)-2-CYclohexyl--l-[(R~-hydroxy(lH--lmidazol-2 vl)methyl]ethyl]-N2-[N-(pyrrolidinYlcarbonyl)-L
~henylalanyl]~L-histidinamide, dihydrochloride a) N-~1-Pyrrolidinylcarbonyl)-L-phenylalanine, methvl ester N-Methylmorpholine (11 ml., 100 mmole) and phosgene (101 ml. of 12% solution in benzen~, 80 mmole) are added rapidly (dropwise) to a solution of pyrrolidine (3.34 ml., 40 mmole) in methylene chloride (200 ml.) at -30 under argon.
The resulting mixture is stirred for one hour at -30, then for one hour as the temperature warms to ~5, after which the mixture is concentrated ln vacuo at 25. The residue is redissolved in methylene chloride. N-Methylmorpholine (13.2 ml., 120 mmole) followed by L-phenylalanine, methyl ester, hydrochloride (8.63 g., 40 mmole) are then added. The mixture is stirred overnight under argon at 25, after which it is concentrated to dryness. The residue is dissolved in ethyl acetate, washed sequentially with water, lN
hydrochloric acid, and saturated aqueous sodium bicarbonate solution, dried (MgS04), and concenta-~5 ted. The residue is chromatographed on silica gel(Merck, 300 g.) eluting with benzene:acetic acid (6:1). Fractions containing the product (Rf =
0.4) are combined and concentrated. The residue (1.5 g.) is crystallized from ethyl acetate/hexane to give 1.19 g. of N-(1-pyrrolidinylcarbonyl)-L-phenylalanine, methyl ester; m.p. 93 - 95;

~ 3 ~ 0792 HA375a [~]D = -19.4 (c = 1, methanol~.
b) N-(1-Pyrrolidinylcarbonyl?~L-E~henylalanine A mixture of the methyl ester product from part ~a) (1.187 g., 4.3 mmole),aqueous lN sodium hydroxide solution (5.15 ml., 5.15 mmole), and methanol (17 ml.? is stirred at 25 for 4 hours, after which it is concentrated ln vacuo. lN
Hydrochloric acid and ethyl acetate are added to the residue, and the mixture is extracted with ethyl acetate. The extract is dried (MySO4) and concentrated to gi~e N-(l-pyrrolidinylcarbonyl)-L-phenylalanine; [~]D = -12.8 (c = 1, methanol).
c) (~R,~S)-~ Am ~ };IIE~hen lmethoxy)me~hyl]-lH~imidazol-2-yl]cyclohexanepro~anol A solution of [(lS)-l-(cyclohexylmethyl)-2-hydroxy-2-[1-[(phenylmethoxy)methyl]-lH-imidazol-~-yl]ethyl]carbamic acid, 1,1-dimethylethyl ester (slow moving isomer) (3.92, 8.83 mmole) [prepared as described in Example 6(d)] in ethyl acetate (200 ml.) is cooled to 0 and HCl gas is bubbled through the solution for 30 minutes. The mixture is then stirred or 3.5 hours as it warms to room temperature, after which it is concentrated ln vacuo to give 3.56 g. of (aR,~S~-~-amino-~-[1-[(phenylmethoxy)methyl]-lH-imidazol-2-yl]cyclohexane-propanol as a white powder.
d) [(l,1-Dimeth~lethoxy?carbonyl~-N-[(lS,2R)-l-(cyclohexylmethyl)-2-hydroxy-2-[1- r ( phenylmethoxy?-methYl]-lH-imidazol-2-yl]-N3-[(phenylmethoxy?methyl]-30 L-histidinamide -~
Triethylamine (2.06 ml., 14.7 mmole) and dicyclohexylcarbodiimide (1.52 g., 7.35 mmole) ~ 3 1 07~2 HA375a are added to a solution of the product from part (c) (3.06 g., 7.35 mmole), l-hydroxybenzotriazole hydrate (1.13 g., 7.35 mmole) and N-(l,l~dimethyl-ethoxy)carbonyl]-l-[(phenylmethoxy)methyl] L-histidine (2.76 g., 7.35 mmole) in tetrahydrofuran (20 ml.). The mixture is stirred for 18 hours at 25, after which it is filtered. The filtrate is diluted with ethyl acetate, washed with saturated sodium bicarbonate solution, dried (MgS04), and concentxated. The residue (4.92 g.) is chromatographed on silica gel (Merck) eluting ~`
with ethyl acetate:pyridine:acetic acid:water (80:20.6:11) to give as the major product 3.98 g.
of [~l,l-dimethylethoxy)carbonyl~-N-[(lS,2R)-l-(cyclohexylmethyl)-2-hydroxy 2-[1-[(phenylmethoxy)-methyl]-lH-imidazol-2-yl]-N3-[(phenylmethoxy)methyl]-L-histidinamide; [~]D ~ -6.1 (c = 1.8, methanol).
e) N-[(lS,2R)-l-(Cyclohexylmethyl)-2-hydroxy-2-[l-[(phenylmethoxy)methyl]-lH-imidazol-2-yl]ethyll-N3-[(phenylmethoxy)methyl]-L-histidinamide A solution of the product form part (d) (3.88 g., 5.53 mmole) in ethyl acetate (200 ml.) is cooled to 0 in an ice bath and HCl gas is bubbled through the solution for 30 minutes. The resulting mixture is then stirred for 2.5 hours as it warms to 25, after which it is concentrated to a small volume. The resulting white precipitate is collected to give 3.33 g. of N-[(lS,2R)-l~(cyclo-hexylmethyl)-2-hydroxy-2-[1-[(phenylmethoxy~methyl]-lH-imidazol-2-yl]ethyl]-N3~[(phenylmethoxy)methyl]-L-histidinamide as a white powder;

1 3 1 0 7~2 HA375a m.p. 143 ~ 157; [~]D = +18-4 (c = 1.0, methanol).
f) N-[(S)-2-Cyclohexyl-l-[(R~hydroxy-[l-[(ph~nylmethoxv)methyl]~lH-imidazol-2~yl]methyl]-ethyl]-N2-[N-(pyrrolidinYlcarbonyl)~L-phenYlalanyl]
N -[(phenylmethoxy)methyl]-L-histidinamide Triethylamine (0.84 ml., 6.0 mmole) followed by dicyclohexylcarbodiimide (453 mg., 2.2 mmole) are added to a mix~ure of N~ pyrrolidinylcar-bonyl)-L-phenylalanine (577 mg., 2.2 mmole), l-hydroxybenzotriazole hydrate (337 mg., 2.2 mole);
and the L-histidinamide product from part (e) (1.47 g., 2.0 mmole) in tetrahydrofuran (8 ml.).
The resulting mixture is stirred overnight as it warms to 25. It is then filtered and the filtrate is diluted with ethyl acetate, washed with saturated sodium bicarbosate solution, dried (MgSO4~, and concentrated. The residue (1;7 g.) is chromatographed onisilica gel eluting with ethyl acetate:pyridine:acetic acid:water (30:20:6:11) to give as the major product 1.46 g.
of N-[(S)-~-cyclohexyl-l-[(R~-hydroxy-[l-~(phenyl-methoxy)methyl]-lH-imida~ol-2-yl]methyl]ethyl~-N2-[N-(pyrrolidinylcarbonyl)-L-phenylalanyl]-N3-[(phenyl-methoxy)methyl]-L-histidinamide; [~]D = ~14.2 (c=0.8, methanol).
g) N-[(S)-2-Cyclohexyl-1-[(R)-hvdroxy-(lH-imidazol-~-yl~methyl]ethyl]-N2-[N-(pyrrolidinYlcarbonyl~-L-henylalanyl~-L-histidinamide, dihydrochloride A mixture of the product from part (f) (1.41 g., 1.60 mmole), lN hydrochloric acid (3.36 ml., 1 3 1 07q2 HA375a 3.36 mmole), and 20% palladium hydroxide on carbon catalyst (300 mg.) in methanol (25 ml.) is stirred under a stream of hydrogen for 24 hours. It is then filtered and concentrated. The residue is lyophillized from water to give 1.0 g. of N-[(S)-2-cyclohexyl-1-[~R)-hydroxy(lH-imidazol-2-yl)-methyl]ethyl]-N2-[N-(pyrrolidinylcarbonyl)-L-phenylalanyl]-L-histidinamide, dihydrochloride as a fluffy white powder; m.p. (168) 175 - 180;
[~]D = -44.8 (c = 0.9, methanol). TLC (silica gel; ethyl acetate:pyridine:acetic acid:water, 40:20:6:11) R~ = 0.28.
32 44 8 4 2.2HC1 3.5 H20:
C, 51.38; H, 7.17; N, 14.98; Cl, 10.43 Found: C, 51.18; H, 7.09; N, 14.98; Cl, 10.48.
Exam~le 12 N-t(S)-2-Cyclohexyl-l-r(R)-hydroxy(lH-imidazol-2-vl)methyllethyl]-N2-[N-~ dimethylethyl)amino]
carbonyll-L-~henylalanvl]-L-histidinamide, dihydrochloride a) N-[(4-Nitrophenoxy~carbonyl]-L~henylalanine, methyl ester N-Methylmorpholine (2.2 ml., 20 mmole) followed by 4-nitrophenyl chloroformate (2.01 g., 10 mmole) are added to a suspension of L-phenyl-alanine, methyl ester, hydrochloride (2.15 g., 10 mmole) in methylene chloride (40 ml.) at -30.
The resulting mixture is stirred at -30 for 15 minutes, then for 15 minutes at 25, after which it is washed sequentially with lN HCl and saturated aqueous sodium bicarbonate solution, 13~01q2 HA375a dried, and concentrated. The residue (2.96 g.) is crystallized from acetonitrile to give 1.22 g. of N-[(4-nitrophenoxy)carbonyl]-L-phenylalanine, methyl ester; m.p. 130 - 131, [~]D = ~88 (c = 1.5, chloroform). Tlle mother liquor is chromatographed on silica gel (90 g.) eluting with benzene:ethyl acetate (9:1) to give an additional 760 mg. of product.
b) N-[[~l,l-Dimethylethyl)aminolcarbonYll-L-phenylalanine, methyl ester _.
l,l-Dimethylethyl amine (0.56 ml., 5.4 mmole) is added to a solution of the product from part (a) (1.48 g., 4.3 mmole) in toluene (21 ml.) at 0.
The resulting mixture is stirred for 24 hours as it warms to 25, after which it is concentrated ln vacuo. The residue is dissolved in ethyl acetate and the solution is washed sequentially with lN
HCl solution, saturated aqueous sodium bicarbonate solution, and saturated potassium carbonate solution. The organic phase is filtered and the filtrate is washed with aqueous potassium carbonàte solution until the washes are colorless.
The or~anic extract is dried (MgSO4) and concentrated in vacuo. The residue (1 17 g.) is ~5 crystallized from ethyl acetate/hexane to give 850 mg. of N-[[(l,l-dimethylethyl~amino]carbonyl]-L-phenylalanine, methyl ester; m.p. 84-86;
[~]D = +24.4 (c = 0.9, methanol).
c) N-[[~ Dimet~y~l Phenvlalanine A mixture of the methyl ester product from part (b) (838 mg., 3.0 mmole) and aqueous lN

I ~ 1 07q2 HA375a sodium hydroxide solution (3.3 ml., 3.3 mmole) in methanol (3 ml.) is stirred for 2 hours at 25, after which it is concentrated in vacuo. The residue is dissolved in water and washed with ethyl acetate. The agueous layer is made acidic by the addition of lN HCl solution and extracted with ethyl acetate. The extract is dried (MgSO4) and concentrated to give 603 mg. of N-[[(l,1-dimethylethyl)amino]carbonyl]-L-phenylalanine.
[a]D = +39.6 (c = 0.7, methanol).
d) N-r(S)-2-C~clohexyl-1-[(R)-hydroxy[l-[(phenyl-~-methoxy~methvl~ imidazol-2-y~lmethyl]ethyl]-N2-[N-rU~ dimethylethyl2a ino~c rbonyll L-phenyl-alanvl]-N3-~phenylmethoxy~methyl]-L-histidinamlde Triethylamine (0.84 ml., 6.0 mmole) and dicyclohexylcarbodiimide (453 mg., 2.~0 mmole) are added to a mixture of N-[(lS,2R)-l-(cyclohexyl-methyl)-2-hydroxy-2-[1-[(phenylmethoxy)methyl]-lH-imidazol-2-yl]ethyl]-N3-[(phenylmethoxy)methyl]-L-histidinamide (1.47 g., 2.0 mmole) [prepared as set forth in Example ll(e)], the product from part (c) (582 mg., 2.20 mmole), and l~hydroxybenzotriazole hydrate (337 mg., 2.20 mmole) in tetrahydrofuran (8 ml.) at 0. The resulting mixture is stirred ~5 overnight as it warms to 25, after which it is filtered. The filtrate is diluted with ethyl acetate, washed with saturated sodium bicarbonate solution and brine, dried (~gSO4), and concentrated.
The residue is flash chromatographed on silica gel (Merck) eluting with ethyl acetate:pyridine:acetic acid:water (80:20:6:11) to give as the major ~ 3 1 0792 ~76- HA375a product 1.48 g. of N-[(S)-2-cyclohexyl-l~[(R)-hydroxy[l-[(phenylmethoxy)methyl] lH-imidazol-2-yl]methyl]ethyl]-N2-[N-[~(l,l-dimethylethyl)amino]-carbonyl]-L-phenylalanyl]-N3-[(phenylmethoxy)methyl]-L-histidinamide. [~]D = ~3-5 (c = 9, methanol).
e) N-[(S)-2-CyclohexYl-l-[(R)-hYdroxY(lH-imidazol-2-vl)methyl]ethyll-N2-[N-L[(l,l-dimethylethyl)amino]
carbonyl]-L-phe~ylalanyl~-L-histidinamide, dihydrochloride A mixture of the product from part (d) ~-(1.43 g., 1.69 mmole) and 20% palladium hydroxide on carbon catalyst (300 mg.) in methanol (25 ml.) is stirred under a stream of hydrogen for 24 hours, after which it is filtered and concentrated. The residue is lyophilized from water to give 1.064 g.
of N-[(S)-2-cyclohexyl-1-[(R)-hydroxy(lH-imidazol-2-yl)methyl)ethyl]-N2-[N-[[(l,l-dimethylethyl)amino~-carbonyl]-L-phenylalanyl]-L-histidinamide, dihydro-chloride as a white solid; m.p. tl75) 178-185;
[~]D = -27.7~ (c = 0.9, methanol). TLC (silica gel;
ethyl acetate:pyridine:acetic acid:water, 40:20:6:11) Rf = 0.27.
32 46 8 4 2.lHC1 2.5~2O:
C, 52.77; H, 7.35; N, 15.38; Cl, 10.22 Found: C, 52.56; ~, 7.36; N, 15.24; Cl, 10.37.

1 3 1 07q2 HA375a Example 13 N-~(S)-2-Cyclohexyl-l-[(R)-hydroxy(lH-imidazol-2-yl~methyllethyll-N2-[N-(cyclopentylcarbonyl~-L-phenylalanyll-L-histidinamide~ dihydrochloride a) N -(L-Phenylalanyl)-N-[(lS,2R)-2~cyclohexyl-1-[hydxoxy[1~ enylmethoxy)methyll~lH-imidazol-2-yl methyl]ethyl~-N3-l~h nYlmethoxY?methyl]-L-histidin-amide, trihydrochloride salt A solution of N2-[N-[(1,1-dimethylethoxy)-carbonyl]-L-phenylalanyl]-N-[(S)-1-[(R)-hydroxy-[l-(phenylmethoxy)methyl] lH-imidazole-2-yl]- --methyl]-2-cyclohexyleth~1]-3'-[~phenylmethoxy)-methyl]-L-histidinamide (7.63 g., 9.0 mmole~
[prepared as set forth in Example 7(a)] in ethyl acetate (325 ml.) is cooled in an ice-water bath under argon and then saturated with HCl gas. The mixture is stoppered and stirred cold for 30 minutes, then the bath is removed and the mixture is allowed to warm to 25 over 60 minutes.
Removal of the solvents ln vacuo followed by drying of the colorless solid product in vacuo gives 7.64 g. of crude N2-(L-phenylalanyl)-N-[(lS,2R)-2-cyclohexyl-1-[hydroxy[1-[(phenylmethoxy)-methyl]-lH-imidazol-~-yl]methyl]ethyl]-N3-[(phenyl~
~S mathoxy)methyl]-L-histidinamide as a trihydrochloride salt.
b) N-[(S)-2-Cyclohexyl-I- r ( R)-hydroxy[l- r ( phenyl-methoxy)methyl]-lH-imidazol-2-y~methyl]ethyl]-N2-~N-(cyclo~entylcarbonyl)-L-phenYlalanYl]-N3-[(Phenylmethoxy)methyl]-L-histidinamide Triethylamine (0.63 ml., 4.5 mmole) and dicyclohexylcarbodiimide (340 mg., 1.65 mmole) 1 3 1 ~7q2 HA375a are added to a solution of the trihydrochloride salt product from part (a) (1.34 g., 1~5 mmole), l-hydroxybenzotriazole hydrate (252 mg., 1.65 mmole), and cyclopentanecarboxylic acid (0.18 ml., 1.65 mmole). The resulting mixture is stirred for 18 hours at 25 after which it is filtered. The filtrate is diluted with ethyl acetate, washed with saturated sodium bicarbonate solution, dried (MgSO4), and concentrated. The residue (1.25 g.) is chromatographed on silica gel (Merck) eluting with ethyl acetate:pyridine:acetic acid:water (80:20:6:11) to give as the major product 800 mg.
of N-[(S)-2-cyclohexyl-l-[(R)-hydroxy[l-[(phenyl-methoxy)methyl]-lH-imidazol-2~yl]methyl]ethyl]-N2-[N-(cyclopentylcarbonyl)-L-phenylalanyl]-N3-[(phenyl-methoxy)methyl]-L-histidinamide [~]D = -8.1 (c = 1.04, methanol).
c) N-[(S~-2-Cyclohexyl-l-~(R)-hydroxy(lH-imidazol-2-yl)methyl]ethyl]-N -~N-(cyclopentylcarbonyl)-L 0 PhenYlalanvlL-_-hlstidinamide, dihydrochloride A mixture of the product from part (b) (740 mg., 0.87 mmole), 20% palladium hydroxide on carbon catalyst (150 mg.), and 1.0 N hydrochloric acid (1.83 ml., 1.83 mmole) in methanol (30 ml.) ~5 is hydrogenated under a slow stream of hydrogen ~or 18 hours. The mixture is then filtered and concentrated to dryness. The residue is dissolved in water and lyophilized to give N-[(S)-2-cyclohexyl-l-[(R)-hydroxy(lH-imidazol-2-yl)methyl]ethyl]-N2 [N-(cyclopentylcarbonyl)-L-phenylalanyl]-L-histidin-amide, dihydrochloride; m.p. (155) 174-177.

1 3 1 07~2 HA375a ~]D = -28.2~ (C = 0.97, methanol). TLC (silica gel; ethyl acetate:pyridine:ac~tic acid:water, 40:20:6:11) Rf = 0.2S.
Anal- calc'd- for C33H45N7Q4 2-15 HCl 3-36 H20 C, 53.37; H, 7.31; N, 13.20; Cl, 10.26 Found: C, 53.37; H, 7.30; N, 13.31; Cl, 10.31.
Example 14 N-~(S)-2-Cyclohexyl-1-_[~R)-hydrox ~lH- midazol-?-yl)methyllethYl]-N -~N-(3,3-dimethvl-1-oxobutyl)-L-phenylalanyl]-L-histidinamide, dihy~_chloride a) N- r (S~-2 Cyclohexyl-l-~(R)-hydroxy[l-[(phenvl-.
methoxY)methy~-lH-imidazol-2-ylLmethyllethyll-N2-~N-(3,3-dimethyl-1-oxobu~yl)-L-phenylalanyl]-N3-[(phenvlmethoxy)methyll-L-histidinamide Triethylamine (0.63 ml., 4.5 mmole) followed by dicyclohexylcarbodiimide (340 mg., 1.65 mmole~
are added to a solution of N2-(L-phenylalanyl)-N-[(lS,2R)-2-cyclohexyl-1-[hydroxy[l-[(phenylmethoxy)-methyl]-lH-imidazol-2-yl]methyl]ethyl]-N3-[~phenyl-methoxy)methyl]-L-histidinamide, hydrochloride salt (1.34 g., 1.5 mmole) [prepared as described in Example 13(e)], l-hydroxybenzotriazole hydrate (252 mg., 1.65 mmole~ and 3,3-dimethylbutanoic acid (0.21 ml., 1.65 mmole) in tetrahydrofuran (5 ml.) at 0. The resulting mixture is stirred for 18 hours as it warms to 25, after which it is filtered. The filtrate is diluted with ethyl acetate, washed with saturated aqueous sodium ~ bicarbonate solution, dried (MgS04), and concentrated. The residua (1.23 g.) is purified by flash chromatography on silica gel (Merck, 150 g.) 1 ~ 1 07q2 HA375a --~0--eluting with ethyl acetate:pyridine:acetic acid:
water (80:20:6:11) to give as the major product 730 mg. of N-[(5)~2-cyclohexyl-1-[(R)-hydroxy[l-[(phenylmethoxy)methyl]-lH-imidazol-2-yl]methyl]-ethyl]-N -[N-(3,3-dimethyl~l-oxobutyl)-L-phenyl-alanyl]-N3-[(phenylmethoxy)methyl]-L-histidinamide;
[~]D = -11.5 (c=l, methanol).
b) N-[5S)-2-Cyclohexyl-l-L(RI-h~droxy(lH-imidazol-2-vl)methylL~thYl]-N2-[N-(3,3-dimethyl-1-oxobutyl)-0 L-phenylalanyl]-L-histidinamide~ dihydrochloride A mixture of the product from part (a) (660 mg., 0.8 mmole), 20% palladium hydroxide on carbon catalyst (150 mg.) and 1.0 N hydrochloric acid (1.7 ml., 1.7 mmole) in methanol (20 ml.) is hydrogenated under a slow stream of hydrogen for 24 hours. The mixture is then filtered and concentrated to dryness. The residue (540 mg.) is dissolved in water and activated charcoal (50 mg.) is added. The resulting mixture is filtered and lyophillized to give N-[(S)-2-cyclohexyl-1-[(R)-hydroxy(lH-imida7ol-2-yl)methyl]ethyl]-N2-[N-(3,3 dimethyl-l-oxobutyl)-L-phenylalanyl]-L-histidin-amide, dihydrochloride; m.p. 158~184~;
[~]D = -32.4~ (c = 0.79, methanol). TLC (silica ~el; ethyl acetate:pyridine:acetic acid:water, 40:20:6:11) R~ = 0.27.
r C33H47N704 2.15 HCl 2.5 H20:
C, 54.35; H, 7.4~; N, 13.45; Cl, 10.45 Found: C, 54.16; H, 7.45; N, 13.59; Cl, 10.42.

~ 3 1 ~7~1~

HA375a Example 15 (lS,2R) N-~l-(CvclohexylmethYl~-2-hydro2y-2-51H-imidazol-~-yl~ethvl]-N2-~N-(4 morpholinylcarbonyl)-L-~henvlalanyl]-L-histidinamide, trifluoroacetate salt a) N-(4-Morpholinylcarbonyl)-L-phenvlalanine, methyl ester Morpholine (1.1 ml., 12.5 ml.) is added to a solution of N-[(4-nitrophenoxy)carbonyl]-L-phenyl-alanine, methyl ester (3.44 g., 10 mmole). The resulting mixture is stirred for 2 hours at 25, then at 100 for 5 hours, after which it is concen-trated ln vacuo. The residue is dissolved in ethyl acetate and the solution is washed with saturated potassium carbonate solution until the washes are colorless. The organic extract is dried (MgS04) and concentrated ln vacuo. The residue is crystallized from ethyl acetate/hexane to give 2.3 g. of N-(4-morpholinylcarbonyl)-L-phenylalanine, methyl ester; m.p. 88 - 91;
[a]D = -30.8 (c = 0.6, methanol).
b) N-(4-Moroholinylcarb n ~ ~-L-phenylalanine A mixture of the methyl ester product from part ~a) (2.3 g., 7.8 mmPle) and agueous lN sodium hydroxide solution (8.6 ml., 8.6 mmole) in methanol (12 ml.~ is stirred for 5 hours at 25, after which it is concentrated ln vacuo. The residue i5 dissolved in water and washed with ethyl acetate.
The extract is dried (MgS04) and concentrated to give 2.2 g. of N-(4-morpholinylcarbonyl)-L-phenyl-~ alanine; [~]D = -23.8 (c = 2, methanol).

1 3 1 o 7 r~ L

HA375a c) ~lS,2R)-N-[1-(Cyclohexylmethyl)-2-hydroxy-2-[1-~(phenylmethoxy)methyl]-lH-imidazol-2-yll-N
[N-(4-morpholinylcarb n~-L-~henylalanyl]-N3-[(phenylmethoxy)methyl]-L-histidinamide Triethylamine (0.84 ml., 6.0 mmole) and dicyclohexylcarbodiimide (435 mg., 2.20 mmole) are added to a mixture of N-C(lS,2R)-l-(cyclo-hexylmethyl)-2-hydroxy-2-[1-[(phenylmethoxy)-methyl]-lH-imidazol-2-yl~ethyl]-N3-[(phenyl-methoxy)methyl]-L-histidinamide (1.49 g., 2.0 mmole) [prepared as set forth in Example ll(e)], the product from part (b) ~612 mg., 2.20 mmole), and l-hydroxybenzotriazole hydrate (337 mg., 2.20 mmole) in tetrahydrofuran (8 ml.) at 0.
lS The resulting mixture is stirred for 18 hours as it warms to 25, after which it is filtered. The filtrate is diluted with ethyl acetate, washed with saturated sodium bicarbonate solution and brine, dried (MgSO4), and concentrated. The residue is flash chromatographed on silica gel (Merck) eluting with ethyl acetate:pyridine:acetic acid:water (100:20:6:11) to give as the major product 1.42 g. of (lS,2R)-N-[1-(cyclohexylmethyl)-2-hydroxy-2-[1-[(phenylmethoxy)methyl]-lH-imidazol-2-yl]-N2-[N-(4-morpholinylcarbonyl)-L-phenylalanyl]-N3-[(phenylmethoxy)methyl]-L-histidinamide;
[~]D = -15.3 (c = 0.9, methanol).
d) (lS,2R~-N-[l-(CYclohexYlmethyl)-2-hvdroxy-2-(lH-imidazol-2-yl)ethyl]-N -IN-(4-mor~holinylcar-bonyl)-L-Phenylalanyll-L-histidinamide, trifluoro-acetate salt A mixture of the product from part (c) (1.4 g., 1.6 mmole), 1.0 N hydrochloric acid 1 3 1 0 7, 2 HA375a ~3.55 ml., 3.55 mmole), and 20% palladium hydroxide on carbon catalys~ ~300 mg.) in methanol (25 ml.) is stirxed under a stream of hydrogen for 18 hours, after which it is filtered and concentrated. The residue (1.7 g.) is purified by preparative HPLC
~YMC S15 ODS column 20 x 500 mm., 25 ml/min of 56% agueous methanol containing 1~ trifluoroacetic acid, W absorbance monitored at 215 nm.).
Fractions containins the major product (retention 1~ time 22 minutes) are combined and concentrated.
The residue is lyophilized from water to give 850 mg.
o~ (lS,2R)-N~[l-~cyclohexylmethyl)-2-hydroxy-2-(lH-imidazol-2-yl)ethyl]-N2-[N-(4-morpholinyl~
carbonyl)-L-phenylalanyl]-L-histidinamide, trifluoro-lS acatate salt as a white solid; m.p. (76) 89-112;
[~]D = -38.1 (c = 0.935, methanol). TLC (silica gel; ethyl acetate:pyridine:acetic acid:water, 40:20:6:11) Rf = 0.19.
~nal. calc'd. for C32H44N8O5 2.2 C2 3 2 2 ~0 C, 48.65; H, 5.52; N, 12.47; F, 13.95 Found C, 48.69; H, 5.60; N, 12.49; F, 14.09.
Exam~le 16 (lS,2R~-N2-~N-~(4-Methyl~ erazinyl)carbonylL-L-~henylalanylL-N-tl-(cyclohexylmethyl)-2-h~droxy-2-S ~lH-imida201-2-yl)ethyll-L-histidinamide, trihydrochloride a) N-r(4-Methyl~ zinyl)carbonyl]-L
henvlalanine, methyl ester A solution of N-[(4-nitrophenoxy)carbonyl]-L-phenylalanine, methyl ester (3.44 g., 10 mmole) in toluene (40 ml.) is heated to reflux and l-methyl HA375a piperazine (1.4 ml., 12.5 mmole) is added to the warm solution. The mixture is stirred for 3 hours as it cools to 25, after which it is concentrated ln vacuo. The residue is dissolved in ethyl acetate and washed with aqueous potassium carbonate solution. The residue (3.85 g.) is crystallized from ethyl acetate to give 2.33 g. of N-[(4-methyl-1-piperazinyl)carbonyl]-L-phenylalanine, methyl ester as an off white solid; m.p. 137 - 138~;
[~]D = ~35 9 (c = 1, methanol).
b) N-~(4-Methyl-l-Diperazin~l)carbonyll-L-phenylalanine A solution of the methyl ester product from part (a) (2.29 g., 7.5 mmole) in methanol (12 ml.) and 1.0 N sodium hydroxide solution (8.25 ml., 8.25 mmole) is stirred for 3 hours at 25, after which the methanol is removed ~n vacuo. The residue is acidified by the addition of excess 1.0 N ~Cl solution and is applied to a cationic exchange column (lO0 ml bed of AG 50 W-X2). The column is eluted with water until the eluant is no longer acidic and then is eluted with 2% aqueous pyridine. Product containing fractions are pooled and concentrated. The residue tl.0 g.) is crystal-lized by trituration with refluxing ethyl acetateto give 860 mg. of N-[(4-methyl-1-piperazinyl)car-bonyl]-L-phenylalanine as a crystalline solid;
m.p. 130 - 132.

l3107Q2 HA375a c) (lS,2R)-N2~[N-~(4-MethYl-1-piperazinyl)car-bonyl]-L-phenylalanyll-N-[l-(cyclohexYlmethyl)-2-hydroxy-2-~ (phenylmethoxy)methyl~-lH-imida ~-vllethv~l]-N -[(~henylmethoxy~methyl]-L-histidin-amide Triethylamine (0.92 ml., 6.6 mmole) and dicyclohexylcarbodiimide (453 mg., 2.2 mmole) are added to a solution of N-[(lS,2R)-l-(cyclohexyl-methyl)-2-hydroxy-2-[1-[(phenylmethoxy)methyl]-lH-imida2O1-2-yl~ethyl]-N3-[(phenylmethoxy)methyl]-L-histidinamide (1.5 g., 2~0 mmole) [prepared as -.-described in Example ll(e)~, l-hydroxybenzotriaæole hydrate (337 mg., 2.2 mmole), and the product from part (b) (690 mg., 2.2 mmole) in dimethylfo~mamide (8 ml.) at 0. The mixture is stirred for 18 hours at 25, after which it is filtered. Ethyl acetate is added to the filtrate and the mixture is washed with saturated aqueous sodium bicarbonate solution and brine, dried (MgSO4), and concentrated.
~he residue is flash chromatographed on silica gel (150 g., Merck) eluting with chloroform:methanol:
ammonium hydroxide (100:12.5:0.25) to give as the major product 600 mg. of (lS,2R)-N2~[N-[(4-methyl-l-piperazinyl)carbonyl]-L-phenylalanyl]-N-[l-(cyclohexylmethyl~-2-hydroxy-2-[1-~(phenylmethoxy)-methyl]-lH-imida~ol-2-yl]ethyl]-N3-[(phenylmethoxy)-methyl]-L-histidinamide; [~]D = -21.5 (c = 1, methanol).

1 3 ~ 0792 HA375a d) (lS,2R)-N2-[N-[(4-Methyl-l-~iperazinyl)car-bonyl]-L-phenylalanyll-N-[1-(cyclohexylmethyl~-2-_droxy-2-(lH-imidazol-2~yl)ethyl]-L-histidinamide, trihydrochloride A mixture of the product from part (c) (563 mg., 0.64 mmole~, 20% palladium hydroxide on carbon catalyst (125 mg.), l.ON HCl solution (2.12 ml., 2.12 mmole) and methanol (20 ml.) is hydrogenated under a slow stream of hydrogen for ~0 hours, after which it is filtered and concentrated. The residue is dissolved in water, charcoal filtered, and lyophillized to give 442 mg. of (lS,2R)-N -[N~[~4-methyl-1-piperazinyl)-carbonyl]-L-phenylalanyl]-N-[1-(cyclohexylmethyl)-2-hydroxy-2-(lH-imidazol-2-yl)ethyl]-L-histidinamide, trihydrochloride; m.p. 178 - 202; [~]D = ~ 54 4 (c = 0.97, methanol). TLC (silica gel; chloroform:
methanol:ammonium hydroxide, 100:25:1) Rf = 0.39.
33 47 94 3-3 HCl 4.0 ~2 C, 47.96; H, 7.11; N, 15.26; Cl, 14.16 Found: C, 47.96; H, 7.06; N, 15.32; Cl, 14.19.
Example 17 (lS,2Rt-N-rl-(Cyclohexylmethyl)-2-hYdroxy-2-(2-thiazol~l)ethyll-N2-[N-[.(l,l-dimeth~Ylethoxy~car-~5 bonyll-L-phenylalanyll~L hl idinamide, O.5 acetate salt a) (S~-~- r r (l,l-DimethYlethoxY~carbonyl]amino~-cyclohexane ~ acid Platinum oxide catalyst (5 g.) is added to a solution of N-[(l,l-dimethylethoxy)carbonyl]-L-phenylalanine (120 g., 0.452 mole) in absolute 1 3 1 07~2 HA375a ethanol (1 l.). The mixture is placed on a Parr reduction apparatus at 50 lbs. pressure. The absorption of hydxogen is rapid and the hydrogen reservoir needs continued refilling. The reduction proceeds overnight and after 20 hours is completed. The mixture is filtere~ through Celite and concentrated in vacuo to give 124.4 g. of (S)~
~-[[(l,1-dimethylethoxy)carbonyl]amino]cyclohexanepro-panoic acid as a glassy solid colorless xesidue;
[~]D = ~9-5 (c = 1, methanol). TLC (silica gel;
toluene:acetic acid, 4:1) Rf = 0.62.
b) ~S)-~-[[(1,1-Dimethylethoxy)carbon~l~amlno]-N-methox~-N-methvlcvclohexanepropanamide The product from part (a) (22.6 g., 83.3 m~ole) is dissolved in tetrahydrofuran (250 ml.) under a blanket of argon at 26. Carbonyl-diimidazole solid (16.0 g., 98.7 mmole~ is added in portions over one minute. Moderate gas evolution begins shortly after the addition is completed. The mixture remains colorless through-out. ~he mixture is stirred for 30 minutes at 25 during which time it remains clear and colorless.
O,N-Dimethylhydroxylamine hydrochloride (11.5 g., 118 mmole) is then added in a single portion followed immediately by triethylamine (17.5 ml., 125 mmole) in a single portion. Following the triethylamine addition a white precipitate forms.
The mixture is stirred for 3 hours at 25, after which it is poured into lN HCl (400 ml.) and 30 extracted with ether (3 x 200 ml.). The colorless -~
extracts are combined and washed with saturated HA375a so~ium bicarbonate solution (2 x 200 ml.), dried ~MgSO~), and concentrated to give 24.2 g. of (S)-~-[[~l,l-dimethylethoxy)carbonyl]amino]-N-methoxy-N-methylcyclohexaneprQpanamide; [~]D =
-11.1 (c = 7, methanol).
c) (S)-~l-(Cyclohexylmethyl)-2-oxo-2-(7-thia-2 olyl)ethYl]carbamic acid, l,1-dimethylethyl ester A 2 . 6 M solution of n-butyllithium (19.5 ml., 4.78 mmole) is added a solution of thiazole (4.07 g~, 4.78 mmole) in tetrahydrofuran (80 ml.) at -60 undar argon. The reaction is stirred at -60 for 30 minutes. The product from part (b) (7.5 g., 2.4 mmole) in tetrahydrofuran (15 ml.) is added lS dropwise at -60 and the reaction mixture is stirred until the temperature reaches -20 (about 40 minutes). The reaction is quenched with saturated ammonium chloride (40 ml.) and the product is extracted with ether (4 x 200 ml.).
The organic layer is washed with brine, dried (NgSO4), filtèred and concentrated to yield 7.2 g.
of crude product. This material is purified by filtration through a 60 g. pad of Merck silica using a hexane:ethyl acetate (8:2) solvent system.
The ~iltrate is concentrated _ vacuo to yield 6.0 g.
of crystalline (S)-[l-tcyclohexylmethyl)-2-oxo-2-(2-thiazolyl)ethyl]carbamic acid, l,1-dimethylethyl aster; m.p. 64 - 69. TLC (silica gel; hexane:
ethyl acetate, 8:2) Rf = 0.45.
3~ Anal. calc'd. for C17H26N~S3 0~1 hexane C, 60.93; H, 7.90; N, 8~08; S, 9.24 Found: C, 61.17; H, 8.13; N, 7.95; S, 8.97.

1 3 1 ~792 HA375a d) (lS,2R)~ Cvclohexylmethyl~-2-hydroxy-?-(2 ~hiazolyl)ethyllcarbamic acid, ~ dimethyl-ethYl ester The product from part (c) ~2.73 g., 8.07 mmol~) is dissolved in absolute ethanol (50 ml.) and cooled to 5~. Sodium borohydride (0.6 g., 16.14 mmole~ is added portionwise and the reaction mixture is stirred for one hour, diluted with ether (200 ml.), and ~uenched with lN HCl to pH l.
The organic layer is separated, washed twice with water and brine, dried (MgSO4), and concentrated .-n vacuo. The two isomers are separated by flash chromatography on silica gel (Merck, 300 g.) eluting with ethyl acetate:hexane (3:8). The slower moving isomer is identified as the S,S
configuration and the faster moving isomer is identified as (lS,2R)-[1-(cyclohexylmethyl)-2~
hydroxy-2-~2-thiazolyl)ethyl]carbamic acid, l,l-dimethylethyl ester; [~]D = -30.72 (c = 0.55, methanol). TLC (silica gel; ethyl acetate:hexane;
1:1) Rf = 0.70.
e) (aR,~S)-B-Amino-~-(2-thiazolyl)cyclohexanepro-panol, dihydrochloride The product from part (d) (0.8 g., 2.3 mmole) is dissolved in ethyl acetate ~20 ml.) and HCl is bubbled into the solution for lO minutes, after which it is stirred at room temperature for 4 hours. The reaction mixture is concentrated to give (aR,~S)-~-amino-a-(2-thiazolyl)cyclohexane-propanol, dihydrochloride as a white solid.

~ 3 1 07S2 HA375~

f3 N-~N-[(1,1-Dimethvlethoxv)carbonyl]-L-phenyl~
alanyll-1~-(2,~-dinitrophenyl)-L-histidine 2,4-Dinitrofluorobenzene (4.62 ml., 36.74 mmole) is added to a solution of N-[N-[(l,l-dimethylethoxy)carbonyl]-L-phenylalanyl]-L-histidine (12.4 g., 30.8 mmole) in aqueous sodium bicarbonate (lM,82 ml.) and methanol (103 ml.).
After stirring the solution for 2.5 hours, an additional amount of 2,4-dinitrofluorobenzene (0.6 ml., 4.77 mmole) is added. At the end of a total of 4 hours of reaction time, the mixture is acidified with aqueous hydrochloric acid (lN, 51.3 ml.) to a pH of 3.9 and then diluted with water (500 ml.). The separated solid is filtered and washed with water. This solid (16.6 g.) is then crystallized from hot ethyl acetate. Dicyclohexyl-amine (1.9 ml.) is added and the crystallized salt is filtered and the free acid is regenerated by acidification to give 1.008 g. of N-[N-[(l,1-dimethylethoxy)carbonyl]-L-phenylalanyl]-1'-(2,4-dinitrophenyl)-L-histidine; m.p. 180 - 182;
[~]D = +5.2 (c = 1.4, acetic acid).
g) ~lS,2R~ __[1-(CyclohexYlmethYl)-2-hydroxy-2-(2-thiazolyl ? ethyll~N2-rN-r~ m~5hylethoxy)-carbon~l~-L-~henylalanvl]-1'-(2,4-dinitrophenyl~
L-histidinamide .. .. _ N-Methylmorpholine (0.23 ml., 2.1 mmole) is added to a solution of the crude dihydrochloride salt product from part (e~ (0.53 g., 1.5 mmole), N-[N-[(1,1-dimethylethoxy)carbonyl]-L-phenyl-alanyl]-1'-(2,4-dinitrophenyl)-L-histidine HA375a --91~

(O.85 g., 1.5 mmole), and 1 hydroxybenzotriazole hydrate (0.2 g., 1.5 mmole) at 0 under argon.
Dicyclohexylcarbodiimide (0.31 g., 1.5 mmole) is finally added and the reaction is kept at 0 overnight, diluted with ethyl acetate (300 ml.), and filtered. The filtrate is washed with water, s~turated sodium bicarbonate solution, and brine, dried ~gSO4), filtered and concentrated 1n vacuo to yield 1.4 g. of crude product. Chromatography on silica gel (Merck, 200 g.) eluting with methanol:chloroform (5:100) gives 0.8 g. of (lS,2R)-N-[l-(cyclohexylmethyl)-2-hydroxy-2-(2-thiazolyl)ethyl]-N2-[N-[(l,l-dimethylethoxy)-carbonyl]-L-phenylalanyl]-1'-(2,4-dinitrophenyl)-1~ L-histidinamide; [~]D = -15.69 (c = 0.51, methanol). TLC (silica gel; methanol:chloroform,

5:85) Rf = 0.25.
h) (lS,2R)- ~ exvlmet~y~ Ydroxy-2-(2-thlazolyllethyl~-N -[N-~(l,l-dimethylethoxy2-car~g~ lalanyll-L-h~stldinamide, 0.5 acetate salt The product from part (g) (0.8 g., 1.0 mmole) is treated with thiolacetic acid (2.8 ml.) in dimethylformamide (4 ml.) at room temperature.
After 2 hours, the reaction mixture is concentrated ln vacuo, diluted with ethyl acetate (200 ml.) and ether (100 ml.), washed with saturated sodium bicarbonate solution (twice) and brine (twice), dried (MgSO4), and concentrated ln vacuo to give 0.8 g. of crude product. This material is chromatographed on silica gel (Merck, 1 3 1 07q~

HA375a 200 g.) eluting with methanol:chloroform (5:100) to remove the (S,S) isomer contamlnant. The solvent system is changed to chloroform:methanol:
water:acetic acid (90:10:1:0.5) to elute the desired ~S,R) isomer. The solution is concentra~
ted ln vacuo, dissolved in 2% aqueous acetic acid, filtered (millipore), and lyophilliæed to give O . 15 g . of ( 1 S, 2R ) -N- [ 1- ( cyclohexylmethyl)-2-hydro~y-2-(2-thiazolyl)e~hyl]-N2 [N-[(l,l-dimethyl-ethoxy)carbonyl]-L-phenylalanyl]-L-histidinamide, 0.5 acetate salt as a light yellow powder; m.p.
110 - 115; ~]D = -32.75 (c = 0.4, methanol).
TLC (silica gel; methanol:chloroorm, 1:9) Rf = O.29.
Anal. calc'd- for C32H44N65S 0 5 C2~I42 H20 C, 58.90; H, 7.19; N, 12.49; S, 4.76 Found: C, 58.69; H, 6.94; N, 12.38; S, 4.92.
Example 18 N-[(S)-1-~HydroxY(lH~imidazol-4-yl)methyl]-3-methyl~
butvl]-N2-rN-[~-dimethylethoxy)carbonyl]-L-Dhenylalanyl]-L-histidinamide, 0.3 acetate salt a) 1-[(PhenylmethoxY)methyl]-2-(phenylthio)-lH-imidazole A solution of n-butyllithium (3.8 ml., 2.6 N in hexane) is added dropwise to a solution of l-[(phenylmethoxy)methyl]-lH-imidazole (1.88 g., 10 mmole) in tetrahydrofuran (35 ml.) at -60~ under argon. After stirring for 2 hours, this solution is added dropwise to a solution of diphenyldisulfide ~1.88 g., 10 mmole) ~t -60 and stirred at -50 to 30 ~60 for 2 hours. Saturated ammonium chloride `-(20 ml.) is added dropwise at room temperature and ~3~07S2 HA375a the crude product is extracted with ether (2 x 200 ml.), washed with brine, dried ~MgS04) and concentrated in vacuo. Flash chromato~raphY on .
silica gel (120 g., LPS-1) eluting with hexane:
ethyl acetate (7:3) yields 2.2 g. of l-[(phenyl-methoxy)methyl]-2-phenylthio-lH-imidazole. TLC
(silica gel; hexane:ethyl acetate, 7:3) Rf = 0.21.
b) [(l,l-DimethYlethoxy)carbonYl]-N-methoxy-N-methyl-L-leucinamide N-[(l,l-Dimethylethoxy)carbonyl]-L-leucine hydrate (10 g., 40 mmole) is dissolved in excess toluene and concentrated to dryness. The residue is dissolved in tetrahydrofuran (100 ml.) and carbonyldiimida~ole (7.8 g., 48 mmole) is added in a single portion. The mixture is stirred at room temperature for 30 minutes. 0,N-Dimethylhydroxyl-amine hydrochloride (4.3 g., 44 mmole) and txiethyl-amine (6.2 ml., 44 mmole) are then added. The resulting mixture is stirred at room temperature ~0 for 3 hours, after which it is poured into excess lN hydrochloric acid. The mixture is extracted - three times with ethyl acetate and the extracts are washed once with lN hydrochloric acid and twice with saturated sodium bicarbonate solution, ~5 dried (MgSO4), and concentrated to give 9.3 g. of [(l,l-dimethylethoxy)carbonyl]-N-methoxy-N-methyl-L-leucinamide as a colorless oil; [a]~ = -25.2 (c - 1.6, methanol).
c) (S)-r3-Methyl-1-[~3-(phenylmethoxy)methyl]-2-(phenylthio)-3H-imidazol-4 yl]carbonyl]butyl]-carbamic acid, l,l-dimethylethyl ester Lithium dii~opropylamide is prepared 1n 1 3 1 07q2 HA375a --9~L--situ by the dropwise addition of n-butyllithium (3.2 ml., 2.6 N in hexane) to a solution of diisopropylamine (1.2 ml., 8.4 mmole) in tetra-hydrofuran (24 ml.) under argon at -60 for 15 minutes. The product from part (a) (2.36 g., 8 mmole) in tetrahydrofuran (20 ml.) is added dropwise at -50 and stirred at -73 for 10 minutes. The product from part (b) (1.08 g., 3.9 mmole) is added in tetrahydrofuran (6 ml.) at --60 and stirred until the reaction reaches 0 (40 minutes). The reaction is then quenched by -the addition of saturated ammonium chloride (10 ml.) and diluted with ether (400 ml.). The organic layer is separated, washed with saturated ammonium chloride and brine, dried (MgS04), filtered, and concentrated in vacuo. The crude product is purified by flash chromatography on silica gel (480 g., LPS-l) eluting with hexane:
ethyl acetate (8:2) to yield 1.03 g. of (S)-[3-methyl-1-[[3-[(phenylmethoxy)methyl~-2-(phenyl-thio)-3H-imidazol-4-yl]carbonyl]butyl]carbamic acid, i,1-dimethylethyl ester as a yellow solid;
m.p. 86-89. TLC (silica gel; hexane:ethyl acetate, 1:1) Rf = 0.73.
~S Anal. calc'd. for C28H35N3O4S:
C, 65.99; H, 6.92; N, 8.24; S, 6.29 Found: C, 66.19; H, 7.13; N, 8.30; S, 6.45.
d) (S)-[3-Methyl-1-[[3-[(~henyl-e-h-oxy)methyl~ -3H-imidazol-4-yl]carbonyl Lb~ arbamic acid, 1,1-dimethylethyl ester The product from part (c) (0.61 g., 1.02 mmole) is dissolved in methanol (10 ml.) 1 3 1 0 7, 2 HA375a and hydrogenated at atmospheric pressure overnight using 20% palladium hydroxide on carbon (0.5 g.) as cata].yst. The reaction mixture is filtered (Celite), recharged with 0.5 g. of catalyst, and hydrogenated an additional 12 hours. It is then filtered (Celite) and chromatographed on silica gel (60 g., LPS-1) eluting with ethyl acetate:
hexane (2:3). The product containing fractions are concentrated ln vacuo to give 0.38 g. of (S)-[3-methyl-1-[[3-~(phenylmethoxy)methyl]-3H-imidazol-4-yl]carbonyl]butyl]carbamic acid, -.-l,l-dimethylethyl ester. TLC (silica gel; ethyl acetate:hexane, 1:1) Rf = 0.45.
e) (S)~[3-Methyl-1-[~3-[(phenylmethoxy~methYl]-3H-imidazol-4-yl]hYdroxymethvl]butyl]carbamic acid, l,l-dimethylethYl ester A 1 M solution of lithium triethylborohydride (1.6 ml., 1.6 mmole) in tetrahydrofuran is added dropwise, at 0 undr argon, to a solution of the product from part (d) (0.16 g., 0.4 mmole) in tetra-hydxofuran (4 ml.). After 30 minutes, O.S M hydro-chloric acid (2 ml.) is added dropwise at 0, the mixture is diluted with ether (200 ml.) and then washed with water and brine, dried (MgSO4), filtered, and concentrated ln vacuo to give 0.21 g.
of crude product. This material is stirred overnight in 50 ml. of methanol:chloroform (1:1) with 3 g. of Merck silica, filtered, and concentra-ted ln vacuo~ The crude product is chromatographed through silica gel (LPS-1, 100 g.) using a S%
methanol:chloroform solvent system. The product t 3 ~ 0792 HA375a containing fractions are combined and evaporated to give 0.16 g. of (S)-[3-methyl-1-[[3-[(phenyl-methoxy)methyl]-3H-imidazol-4-yl~hydroxymethyl]-butyl]carbamic acid, l,1-dimethylethyl ester as a viscous oil. TLC (silica gel; methanol:chloroform, 1:20) Rf = 0.22.
Anal. calc'd. for C22H23N3O4 H2O
C, 62.70; H, 8.04; N, 9.99 Found: C, 62.70; H, 8.49; N, 9.97.
f) ~-[(S)-l-Amino-3-methylbutyl]~3-L(phenyl-methoxy)methyl]-3H-imidazole-4-methanol A solution of the product from part (e) (0.11 g., 0.26 mmole) in dichloromethane (2 ml.) and trifluoroacetic acid (2 ml.) is stirred at lS 10 for 30 minutes and at room temperature for 10 minutes. The reaction mixture is evaporated ln vacuo and then concentrated from acetonitrile (three times) to give a-[(S)-l-amino-3-methyl-butyl]-3-[(phenylmethoxy)methyl]-3H-imidazole-4-methanol which contains 2.5 M of trifluoro-acetic acid.
Anal. calc'd. for C28H3SN3O4S
C, 44.90; ~, 4.71; N, 7.14 Found: C, 44.50; H, 4.80; N, 7.04 ~S g) N-t(S)-l-rHvdroxyr3-[(~henylmethoxy~methYll-3~-imidazol-4-yl]methylJ-3-methylbutyl]-N2-[N-r(i,l-dimethyleth~y)carbonyl]-L-phenylalanyl N -[(phenylmethoxy)methyl]-L-histidinamide Diisopropylethylamine (0.11 ml., 0.52 mmole) is added to 3 ml. tetrahydrofuran solution of the crude 2.5 M trifluoroacetic acid salt product from 1 3 1 07q2 HA375a part (f) (0.26 mmole), at 0 undr argon, followed by the addition of N-[N-[(l,l-dimethylethoxy)-carbonyl]-L-phenylalanyl]-l'-[(phenylmethoxy)methyl]-L-histidine (135.9 mg., 0.26 mmole), 1-hydroxybenzo-S triazole hydrate (135.2 mg., 0.26 mmole) and finally dicyclohexylcarbodiimide (53.7 mg., 0.26 mmole). After one hour, dimethylaminopyridine (15 mg., 0.12 mmole) is added. ~he reaction mixture is stirred overnight at room temperature, concentrated ln vacuo, dissolved in e~hyl acetat~ .
(150 ml.), and filtered. ~he filtrate is washed with half saturated sodium bicarbonate solution and brine, dried (MgS04), filtered, and concentra-ted i vacuo to give 0.17 g. of crude product. Two flash chromatographies, one on silica gel (Merck, 80 g.) and the other on silica gel (LPS-l, 80 g.
employing the solvent system chloroform:methanol:
ammonia hydroxide (100:10:0.2) yields 60 mg. of N-[(S)-l-[hydroxy[3-[~phenylmethoxy)methyl]-3H-imidazol-4-yl]methyl]-3-methylbutyl]-N2~[N-[(1,1-dimethylethoxy)carbonyl]-L-phenylalanyl]-N3-[(phenylmethoxy)methyl]-L-histidinamide (3:7 isomer ratio).
h) N-~(S)-1-[Hydroxy(lH-imidazol-4-~l)methyl]-3-methylbutyl]-~ ~[N-[(l,1-dimethylethoxy)carbonyll-L-~henylalanyll L-histidinamide, 0.3 acetate salt The product from part (g) (60 mg., 0~74 mmole) is dissolved in methanol t4 ml.), water (0.5 ml.), and lN hydrochloric acid (0.15 ml.) and hydrogenated overnight using 20% palladium hydroxide on carbon catalyst (40 mg.). The ~ 3 ~ 07q2 HA375a reaction mixture is filtered through Celite, concentrated in vacuo, and chromatographed on silica (Merck, 30 g.) eluting with chloroform:
methanol:water:acetic acid ~90:20:2.5:1). The product containing fractions are combined and evaported to yield 27.5 mg. of product. A portion of this sample is dissolved in ethyl acetate and stirred with water. The organic phase is concentrated to yield 12.1 mg. of N [(S)-1-[hydroxy-10 (lH-imidazol-4-yl)methyl]-3-methylbutyl]-N2_[N-[(1~
dimethylethoxy)carbonyl]-L-phenylalanyl]-L-histidinamide, 0.3 acetate salt having an isomer ratio of about 1:4; m.p. 113 - 128. TLC (silica gel; chloroform:
methanol:water:acetic acid, 90:20:2.5:1) Rf = 0.19.
Anal- calc'd- for C~9H41N75 ' 0 3 C2~42 C, 60.70; H, 7.26; N, 16.74 Found: C, 60.80; H, 7.67; N, 16.68.
Example 19 (lS,2R)-N2-[N-~N2-(Cyclobutylcarbonyl~-L-lysyl]-L-phenylalanvll-N-[l-(cyclohexylmethyl)-2-hydroxy-2-(lH-imidazoI-2-yl)ethyll-L-histidlnamide, 3,3-hvdrochloride a) (lS,2R~-N2-~N-LN2-(Cyclobutylcarbonyl)-N6-[(phenylmethoxy)carbonyl~-L-lysYl~-L-~henYlalanyll-N-[l (cyclohexylmethyl)-2-hydroxY~2-~1-[(phenyl-methoxy)methyl~-lH-imidazol-2-vllethyl]-N3-[(phenyl-methoxv)methyl]-L-histidinamide N-Methylmorpholine (586 mg., 5.79 mmole) and dicyclohexylcarbodiimide (398 mg., 1.93 mmole) are added to a mixture of N-[~cyclobutyl)carbonyl]-N6-[~phenylmethoxy)carbonyl]-L-lysine (700 mg., 1 3 1 07~2 HA375a _99_ 1.93 mmole), N2-(L-phenylalanyl)-N-[(lS,2R)-2-cyclohexyl-l-[hydroxy[l-[(phenylmethoxy)methyl]-lH-imida~ol-~-yl]methyl]ethyl]-N3-[(phenylmethoxy)methyl]-L-histidinamide, trihydrochloride salt (1.72 g., 1.~3 mmole) [prepared as described in Example 13(a)], and l-hydroxybenzotriazole hydrate (325 mg., 2~12 mmole) in tetrahydrofuran (20 ml.) at 0~
under argon. The reaction is kept stoppered in a refrigerator for 3 days, then iltered, diluted with ethyl acetate, rinsed with three 15 ml.
portions of water, 15 ml. of saturated sodium .-bicarbonate solution, and brine, dried (MgS04) and concentrated ln vacuo to yield 2.12 g. of crude product. Recrystallization from methanol/
ethyl acetate gives 1.2 g. of (lS,2R)-N2-[N-[N2-(cyclobutylcarbonyl)-N6-~(phenylmethoxy)carbonyl]-L-lysyl]-L-phenylalanyl]-N-[l-(cyclohexylmethyl)-2-hydroxy-2-[1-[~phenylmethoxy)methyl]-lH-imidazol-2-yl]ethyl]-N3-[(phenylmethoxy)methyl]-L-histidinamide;
m.p. 146 - 162 (dec.); [~]D = -14-5 (c = 0.5, methanol).
b) (lS,2R)~N2-[N-[N-_-(CYclobutylcarbonyl)-L-lysyl L-~henylalanyl]-N- r 1-(cyclohexvlmethYl~-2-hydroxy-2-(lH-imidazol-2-yl)ethyl]-L-histidinamide~ 3,3-~5 hvdrochloride A solution containing the product from part (a) (1.05 g., 0.952 mmole), hydrazine hydrate (476 mg., 9.52 mmole), and 20% palladium hydroxide on carbon catalyst (300 mg.) in methanol (50 ml.) is stirred under hydrogen for 24 hours. The reaction mixture is then filtered, concentrated to HA375a remove the methanol, and redissolved in water ~25 ml.) containing lN a~ueous hydrochloric acid ~2.63 ml.). Lyophillization gives 756 mg. of (lS,2R)-N2-[N-[N2-(cyclobutylcarbonyl)-L-lysyl]-L-phenylalanyl]-N-[1-(cyclohexylmethyl)-2-hydroxy-2-(lH-imidazol-2-yl)ethyl]~L-histidinamide, 3.3 hydrochloride; m.p. 174 -187; [~]D = ~30 0 (c - 0.5, methanol). TLC (silica gel; n-butanol:
pyridine:acetic acid:water, 4:1:1:1) Rf = 0.48.
Anal- calc'd- for C38H55N95 3-3 ~Cl 3-3 H2O:
C, 50.84; H, 7.29; N, 14.04; Cl, 13.03 Found: C, 50.84; H, 7.43; N, 14.20; Cl, 12.96.
Example 20 (lS,2~)-N-[l-~Cyclohexylmeth~ hydroxy-2-(lH-imidazol-2-yl)ethyll-N2-[1-oxo-3-phenyl-2-(phenyl~
methyl)propyl~-L-histidinamide, 2.2-trifluoroa etate salt a) 2,2-Bis~phenvlmethvl)propanedioic acid, diethvl ester Diethyl benzylmalonate (8.6 ml., 50 mmole) is added dropwise over 5 minutes to a suspension of sodium hydride (2.0 g., 50 mmole, of 60% dispersion in mineral oil) in tetrahydrofuran (100 ml.). Gas evolution is observed. When the addition is completed, the mixture is heated at reflux for 10 minutes and is then cooled to 25. A solution of benzyl bromide (6.5 ml., 55 mmole) in tetra-hydrofuran (10 ml.) is added dropwise over 10 minutes, after which the mixture is stirred for 20 hours at ?5 under argon. lN Hydrochloric acid (100 ml., 100 mmole) is then added and the mixture 1 3 1 07q2 HA375~

is extracted with e~hyl acetate. Th~ extract is washed with saturated aqueous sodium bicarbonate solution, dried (MgSO4), and concentrated ln vacuo. The residue (17.8 g.) is chromatographed on silica gel (Merck) eluting with hexane:ethyl acetate (5:1) to give 2,2-bis(phenylmethyl)pro-panedioic acid, diethyl ester.
b) ~-(Phenylmethyl)benzenepropanoic acid A solution of the diethyl ester product from part (a) (14.4 g., 4~.5 mmole) in ethanol (lO0 ml.) and 1.0 N aqueous sodium hydroxide solution (95 ml~;
95 mmole) is heated at reflux for 48 hours. The ethanol is removed ln vacuo and ~he remaining aqueous mixture is acidified by the addition of lN
lS hydrochloric acid. The mixture is saturated with sodium chloride and extracted with ethyl acetate.
The extract is dried (MgSO4) and concentrated ln vacuo to give 9.6 g. of the dicarboxylic acid which crystallizes on standing.
A solution of the above material (9.6 g.) in dioxane (100 ml.~ containing concentrated hydrochloric acid (1 ml.) is heated at reflux for 24 hours, after which it is concentrated to dryness. The residue is crystallized from ethyl acetate:hexane to give i.o2 g. of a-(phenylmethyl)-benzenepropanoic acid; m.p. 86 - 87.
c) (lS,2R)-N-[l-(CyclohexylmethYl?-2-hydroxy-2-rl-~(phenylmethoxy)methvl]-lH-imidazol-2-yllethyl]
N2-~l-oxo-3-phenvl-2-(phenYlm thyl)~ropyl]~N3-Ll-(phenylmethoxy)methyl]-L-histidinamide HA375a Triethylamine (0.53 ml., 3.8 mmole) and dicyclohexylcarbodiimide (283 mg., 1.4 mmole) are added to a solution of N-[(lS,2R)-l~(cyclohexyl-methyl)-2-hydroxy-2-[l-[(phenylmethoxy)methyl]-1~-imidazol-2-yl]ethyl] N3-~(phenylmethoxy)methyl]-L-histidinamide (920 mg., 1.25 mmole) [prepared as described in Example ll(e)], l-hydroxybenzotriazole hydrate (210 mg., 1.34 mmole), and the product from part (b) (330 mg., 1.4 mmole) in tetrahydrofuran (5 ml.) at 0. The resulting mixture is stirred for 18 hours at 25, after which it is filtered. The filtrate is diluted with ethyl acetate, washed with saturated sodium bicarbonate solution and brine, dried, and con-centrated. The residue is chromatographed on silica gel (Merck) eluting with ethyl acetate:
pyridine:acetic acid:water (100:20:6:11) to give 900 mg. of (lS,2R)-N-[1-(cyclohexylmethyl)-2-hydroxy~2-[1-[(phenylmethoxy)methyl]-lH-imidazol-2-yl]ethyl]-N2-[1-oxo-3-phenyl-2-(phenylmethyl)propyl]-N3-[(phenylmethoxy)-methyl]-L-histidinamide as the major product;
[~]D = ~5 3~ (c = 1, methanol).
d) (lS,2R)-N-[l-~Cyclohexylmethyl)-2-hydroxy-2-(l~-imidazol-2-vl)ethYl~ [1-oxo-3-phenyl-2-(Phenylmethyl)Propyl]-L-histidinamide, 2,2 trifluoroacetate salt A mixture of the product from part (c) (900 mg., l.l mmole), 20% palladium hydroxide on carbon catalyst (200 mg.), and 1.0 N hydrochloric acid (2~4 ml., 2.4 mmole) in methanol (20 ml.) is 1 3 1 07 ,2 HA375a hydrogenated under a slow stream of hydrogen for 19 hours. The mixture is then filtered and concentrated to dryness. The residue is dissolved in water, activated carbon is added, and the mixture is then millipore filtered and lyophillized to give 590 mg. of a pinkish solid.
This solid is further purified by preparative HPLC (YMC s 15 ODS column, 20 x 500 mm., 67%
aqueous methanol containing 1% trifluoroacetic acid, 25 ml/min., W 220 nm. monitoring).
Fractions containing the major component (retention time 32 minutas) are combined and concentrated. The residue is dissolved in water and lyophillized to give 465 mg. of (lS,2R)-N
[1-(cyclohexylmethyl)-2-hydroxy-2-(lH-imidazol-2-yl)ethyl]-N -[l-oxo-3-phenyl-2-(phenylme~hyl)propyl~-L-histidinamide, 2.2 trifluoroacetate salt as a fluffy white solid; m.p. (70) 78 - 108; [~]D =
-28.0 (c = 0.70, methanol). TLC (silica gel, ethyl acetate:pyridine:acetic acid:water, 40:20:6:11) Rf = 0.36.
Anal. alc d- f r 34 42 6 3 2 3 2 0.8 ~2 C, 54.39; H, 5.4 ; N, 9.91; F, 14.79 Found: C, 54.34; H, 5.55; N, 10.00, F, 14.73.
Example 21 (lS,2R)-N-~l-(Cyclohexylmethyl)-2-hydroxy-2-(lH-imidazol-2-yl)ethyl]-N2-~1-oxo-3-(1-naphthaleny~
2-(1-na~hthalenylme~hyl~propyll-L-histidinamide, d hydrochloride a) 2,2-Bis(l~naphthalenylmethyl~propanedioic acid, diethYl ester Diethylmalonate (15.2 ml., 100 mmole) is added to a suspension of sodium hydride 1 3 1 07~2 HA375a (20Q mmole) in tetrahydrofuran (400 ml.). The addition is accompanied by gas ev~lution. When the addition is completed, ~he mixture is warmed to reflux temperature to yield a homogeneous solution. 1-Chloromethyl naphthalene ~35.4 g., 200 mmole) is added to the refluxing solution over one hour. THe mixture is then stirred at reflux for 17 hours, af~er which it is ~uenched by the addition of excess lN hydrochloric acid. The mixture is extracted with ether and the extract is washed with saturated sodium bicarbonate solution,_ dried (MgSO4), and concentrated to a yellow oil.
The residue is dissolved in hot petroleum ether and the colorless solution is decanted from a brown insoluble gum. The solu~ion is concentrated and the residue is crystallized twice from methanol to give 13g. of 2,2-bis(1-naphthalenyl-methyl)propanedioic acid, diethyl ester as colorless crystals; m. p. 79 - 81.
b) -(l-NaDhthalenylmethyl)-l-na~hthalenyl-~ro~anoic acid A mixture o~ the diethyl ester from part (a) (11.0 g., 25 mmole) and 1.0 N sodium hydroxide solution (50 ml., 50 mmole) in ethanol (50 ml.) is stirred at 25 for 3 days. Sodium hydroxide solid (~ g.) is then added and the mixture is stirred for an additional 6 hours at 25~, after which it is concentrated ln vacuo. The residue is diluted with water, resulting in a dense white precipitate. The precipitate is collectèd and washed with hexane. The filtrate is then washed 1 3~ 07q2 HA375a with hexane and the combine~ hexane washes are extracted with sodium hydroxide Colution. The scdium hydroxide extracts and the solid precipitates are combined and acidified by the addition of concentrated hydrochloric acid. The mixture is then extracted with ethyl acetate and the extract is dried and concentrated. The residue (7.2 g.) is dissolved in dioxane (250 ml.). Concentrated hydrochloric acid (1.0 ml.) is added and the mixture is stirred at 90 for l9 hours, after which it is concentrated ln ' vacuo. The residue is triturated with methanol to give 4.6 g. of ~ naphthalenylmethyl)-1-naphtha-lenylpropanoic acid as a white powder; m.p.
lS 168 ~ 170.
c) N-~(lS,2R)-1-(CyclohexYlmethy~-2-hydroxy-2-(lH-imidazol-2-yl~ethyll-L-histidinamide A mixture of [(l,l-dimethylethoxy)carbonyl]-N-[(lS,2R)-l-(cyclohexylmethyl)-2-hydroxy 2-[1-[(phenylmethoxy)methyl]-1~-imidazol-2-yl]ethyl]-N3-[(phenylmethoxy)methyl]-L-histidinamide (1.46 g., 2.0 mmole) [prepared as set forth in Example 11 (d)], 20% palladium hydroxide on carbon catalyst (350 mg.), and 1.0 N hydrochloric acid (4 ml., 4 mmole) in methanol (15 ml.~ is hydrogenated under a slow stream of hydrogen for 20 hours at 25, after which it is filtered and concentrated to dryness. The residue (1.1 g.) is dissolved in acetic acid l30 ml.) and dry HCl gas is bub~led through the solution for 30 minutes at 25. The mixture is then stirred at 25 for 2 hours, after which is is concentrated 1n vacuo. The residue is 1 3 1 07q2 HA375a triturated with acetonitrile to give 833 mg. of a white solid. The solid is dissolved in excess lN
hydrochloric acid and concentrated to dryness.
The process is repeated three times to give N-[(lS,2R)-1-(cyclohexylmethyl)~2-hydroxy 2-~lH-imidazol~2-yl)ethyl]-L-histidinamide as a white solid.
d) (lS,2R)-N-~-(Cvclohexylmethyl~-2-hydrox~-2-(lH-imidazol-2-yl)ethyll-N2-[1-oxo~3-(1-naphthal-en 1)-2-(1-na~hthalenylmethyl)pr~pyl]-L-histidinamide, dihydrochloride Triethylamine (O.68 ml., 4.9 mmole) and dicyclohexylcarbodiimide (309 mg., 1.5 mmole) are added to a mixture of the product from part (c) (750 mg., 1.5 mmole), 1-hydroxybenzotriazole hydrate (230 mg., 1.5 mmole) and the product from part (b) (511 mg., 1.5 mmole) in dimethylformamide (7 ml.) at 0. The resulting mixture is stirred for 18 hours at 25 after which it is concentrated to dryness. Methanol (9 ml.) and 1.0 N hydro-chloric acid (6 ml.) are added to the residue.
The mixture is filtered and the filtrate is concentrated to dryness. The residue is flash chromatographed on silica gel (Merck) eluting with ethyl acetate:pyridine:acetic acid:water (50:20:6:11) to give a major product (Rf = 0.25) that is homogeneous but dark brown in color. This material is dissolved in l.0 N hydrochloric acid and reconcentrated. The resulting hydrochloride 30 salt (430 mg.) is purified by chromatography on `~
HP-20 eluting with a gradient from 0.01 N hydro-1 3 1 07q2 E~375a chloric acid to methanol. Colorless fractions containing the major product are combined and partially concentrated to remove the methanol. The aqueous residue is lyophillized to give a fluffy electrostatic white solid that is dissolved in water and relyophillized to give 220 mg. of (lS,2R)-N-[1-(cyclohexylmethyl)-2-hydroxy-2 (lH-imidazol-2-yl)ethyl]-N2-[1-oxo-3-(1-naphthalenyl)-2-(1-naphthalenylmethyl)propyl]-L-histidinamide, dihydrochloride; m.p. 163 - 183; [~]D = -56.1 (c = 0.59, methanol). TLC (silica gel; ethyl acetate:pyridine:acetic acid:water, 50:20:6:11) Rf = 0.25.
Anal. calc d- for C42X46 6 3 2 C, 63.06; H, 6.59; N, 10.51; Cl, 9.75 Found: C, 63.03; ~, 6.76; N, 10.61; Cl, 9.78.
~xamples 22 - 46 Following the procedures of Examples 1 to 21, additional compounds within the scope of this invention can be prepared having the forula X-~ NH -CH - C ~ NH - CH- C- N~- CH- CH - R
OH

wherein the substituents are as defined below.

1 3 1 07q2 _ 108- HA375a ~`

z~

~, Q
Q Q Q

z-~ z-~

N N
~ I Q Q

o = ~ o _~
~ y ~ ~ ~ Q
o = V 2: Z
~n O O=y 0= ~

1~ ¦ ` N

~ 3 ~ 07~
HA37 5a 2~ 2 N N
~ N
U V ~
~ ¦ U U

2 [~
N y U

O=c,~ O=U ~=
~_ N I ~_ U -- U '~

O = ~ Z ~ Z
_ =~t O--y 0=~
Z U

O , . ~.
. U~ `D ' ~ , HA37 5a 'o~ ~z =-Z~

~ ~ N
- Q
N N ~
r~ I

Z_-- O ~
y ~ _N ¦ O _ O

$ o=$ o=$ ~
- ~ o ô~ ~

r ' ,~ ~

1 3 1 0.7~2 HA37 5a 0~ z~

N
U ~ ~
~, ~, 3 ~ r~ I U N ~

Z~ _ Z~

O=
O = t.~ l <~3_ I ~N ~ 0 -D

O= Z O=y Z
n_ ~ O Z~ ~
", ~ . Y

~3 ~ .

1 3 1 07q2 HA375a Q ~ ~
~ ~1 u-~
.. ,, I ~ ~ Z = Z

N N N
U U U
U UN N

~ _ 2N~2 ~ I U U U

O =U
U ~ :: _ y O _ U

N I N
t I ~ [~

~ I `, 1 3 ~ 07Q2 HA37 5a u ~ ~ ~ ~u ~`I ~ N
2~ U Q~

U U~ U

~, Z~ Z-~ z N ~`I ~ ~ /
~ I U UU C.) O 0=0 ~ ~ =~ ~

1 3 ~ O/7q2 _ 114- ~1A375a =z~ ~z ~ Z~Z

? ? S' N

O =
O = ~ ~N~
o _ ~, Cl ~:
~ N Z O -- C~ ~
,~ ~ ZO = O O ~ Z

3 ~

1 3 ~ 07q2 ~A375a ~a~
1000 tabl~t~ ~ac~ corltaining th~ followi~g in~redient~ s N2~ lN~ Dim~thyl~thoacy) S casbonyl ] -L-ph~nyl al 3~yl ~ N~
L ( lS ) -2 ~cyclohexyl-l- [ ( R ) -hydro~y~
l~-i;nidazol-2 -ylm~t:hy~ ~ ~thy3 ] -L-histidina~id~, monoacot~ts salt 250 mg.
Cornstarch lOQ mg.
G~lati~ 20 m~.
*~vicol (micxoc:~stalli~o c~llulosel~)50 mg.
~agne~ium stoa~t~
~5 D35~-15 ar~- praparod ~OD~ ~u~ici~nt bullc gu~ti~ y mi~cing th~ active ~nono~c~t~t~ ~lt cO~pOU~la ~d corn~arch w$t~ a~ agu~ouc ~ utlon o~
g~latin. Th~ mixtur~ dl ~d ~o~ ~o ~n3 powd~r. q~ ~Avic~ d th~ t~ ag~
20 st~arat~ ar~ ad~s~d w~th gr~ul~o~O T~L8 mixtus~ i~ th~ comp~e~8~ iDI a t~lbl9'e ~p~081~ to for~ 1000 t~ ~ co~ 25~ ~O o~
activ~
In a 3iln~ oa~er, t~l~t~ co~i~ q 25 2S0 mg. o~ ~ produc~ o~ ~y o~ E:x~splo~ 1 ~o 6 8 ~o 46 ca~ b~ prop&~a~.
_` A ~in~l~ proce~du~o c~ b~ ~ploy~a to ~on~
t~l6t~ co.sl~ai~ 500 laq. o~ ~c~iv~ adi~

* Trademark .

1 3 1 ~7q2 HA375a Exam~le 4~
An injectable solution is prepared as follows:
N -[N-~(1,1-Dimethylethoxy) 5 carbonyl]-L-phenylalanyl]-N-[(S)~1-[(R)-hydxoxy(lH~imidazol-2-yl)methyl]-3-methylbutyl]-L-histidinamide, acetate salt1000 g.
Methyl paraben 5 g.
10 Propyl paraben 1 g.
Sodium chloride 5 g. -.-The active substance, preservatives, and sodium chloride are dissolved in 3 liters of water for injection and then the volume is brought up to 5 liters. The solution is filtered through a sterile filter and aseptically filled into pre-sterilized vials which are closed with presteriliæed rubber closures. Each vial contains 5 ml. of solution in a concentration of 200 mg. of active ingredient per ml. of solution for in] ectlon.
In a similar manner, an injectable solution containing 200 mg. of active ingredient per ml. of solution can be prepared for the product of any of Examples 1, 2, and 4 to 46.

HA375a ExamDle 49 -lO00 tablets each containlng the following ingredients:
N2-[N-[(lll-Dimethyle~hoxy)-S carbonyl]-L-phenylalanyl]-N-[ ( lS ) -2-cyclohexyl-1-[(R)~hydroxy lH-imidazol-2-ylmethyl]ethyl]-L-histidinamide, monoacetate salt 500 mg.
lO Avicel 300 mg.
Hydrochlorothia7ide 14.5 mg.
Lactose 113 mg.
Cornstarch 15.5 mg.
Stearic acid 7 m~.
950 mg.
are prepared from sufficient bulk quantities by slugging the N2-[N-[(l,1-dimethylethoxy)-carbonyl]-L-phenylalanyl]-N-[(lS)-2-cyclohexyl-l-[(R)-hydroxy-lH-imidazol 2-ylmethyl]ethyl]-L-histidinamide, monoacetate salt, Avicel, and a portion of the stearic acid. The slugs are ground and passed through a #2 screen, then mixed with the hydrochlorothiazide, lactose, cornstarch, and remainder of the stearic acid. The mixture ls compressed into 950 mg. capsule shaped tablets in a tablet press. The tablets are scored for dividing in half.
In a similar manner, tablets can be prepared containing 500 mg. of the product of any of Examples l to 6 and 8 to 46.

Claims

HA375a The embodiments of the invention in which an exclusive property or privilege is claimed are defined as follows:

1. A compound of the formula including a pharmaceutically acceptable salt thereof wherein:

X is R6-(CH2)m-, , , R6-(CH)m-O-?-, R6-O-(CH2)n-?- , R6 (CH2)m NH-?- , , R6-(CH2)m-SO2- or HA375a ;

R3, R4, R5 and R10 are independently selected from the group consisting of hydrogen, lower alkyl, halo substituted lower alkyl, -(CH2)n-aryl, -(CH2)n-heterocyclo, (CH2)n-OH, -(CH2)n-O-lower alkyl, -(CH2)n-NH2, -(CH2)n-SH, -(CH2)n-S-lower alkyl, -(CH2)n-O-(CH2)g-OH, -(CH2)n-O-(CH2)g-NH2, -(CH2)n-S-(CH2)g-OH, -(CH2)n-?-OH, -(CH2)n-S-(CH2)g-NH2, and -(CH2)n-cycloalkyl;

HA375a R5 and R6' are independently selected from the group consisting of lower alkyl, cycloalkyl, aryl and heterocyclo;
p is zero or one;
q is zero or one;
m and m' are independently selected from the group consisting zero and an integer from 1 to 5;
n is an integer from 1 to 5;
g is an integer from 2 to 5;

R7 is or R8 is 2,4-dinitrophenyl, or ;

R1 is a fully saturated, partially saturated, or unsaturated monocyclic N-hetero-cyclic ring of 5 or 6 atoms containing at least HA375a one N atom or a bicyclic ring in which said N-heterocyclic ring is fused to a benzene ring wherein said N-heterocyclic ring can also include an O or S atom or up to three additional N-atoms, an available N atom in said N-heterocyclic ring can be substituted with -2,4-dinitrophenyl, lower alkyl, or -(CH2)n-cycloalkyl, an available carbon atom in said monocyclic N-heterocyclic ring or in the benzene portion of said bicyclic N-heterocyclic ring can be substituted with lower alkyl, , or -(CH2)n-cycloalkyl, and said N-heterocyclic ring is bonded to the moiety at an available carbon atom;
the term lower alkyl refers to straight or branched chain radicals having up to seven carbon atoms;
the term cycloalkyl refers to saturated rings of 4 to 7 carbon atoms;
the term halo refers to Cl, Br, and F;
the term halo substituted lower alkyl refers to such lower alkyl groups in which one or more hydrogens have been replaced by chloro, bromo or fluoro groups;

HA375a represents a heterocyclic ring of the formula wherein Y is -CH2, O, S, or N-R9, wherein R9 is hydrogen, lower alkyl, or -(CH2)n-cycloalkyl, a is an integer from 1 to 4, and b is an integer from 1 to 4 provided that the sum of a plus b is an integer from 2 to 5 and such heterocyclic rings wherein one carbon atom has a substituent selected from lower alkyl, lower alkoxy, lowex alkylthio, halo, CF3, nitro, or hydroxy;
the term aryl refers to phenyl, 1-naphthyl, 2-naphthyl, mono subctituted phenyl, 1-naphthyl, or 2-naphthyl wherein said substituent is lower alkyl of 1 to 4 carbons, lower alkylthio of 1 to 4 carbons, lower alkoxy of 1 to 4 carbons, halogen, hydroxy, amino, -NH-alkyl wherein alkyl is of 1 to 4 carbons, or -N(alkyl)2 wherein alkyl is of 1 to 4 carbons, di or tri substituted phenyl, 1-naphthyl or 2-naphthyl wherein said substituents are methyl, mathoxy, methylthio, halogen or hydroxy; and the term heterocyclo refers to fully saturated or unsaturated rings of 5 or 6 atoms containing one ox two O or S atoms and/or one to four N atoms provided that the total number of hetero atoms in HA375a the ring is 4 or less and bicyclic rings wherein the five or six membered ring containing O, S and N atoms as defined above is fused to a benzene ring.

2. A compound of Claim 1 wherein:
R1 is HA375a R2 is 2,4-dinitrophenyl, hydrogen, lower alkyl, , or -(CH2)n-cycloalkyl;

R9 is hydrogen, lower alkyl, , or -(CH2)n-cycloalkyl; and n is an integer from 1 to 5.

3. A compound of Claim 2 wherein:

X is R6-(CH2)m-?-, R6-(CH2)m-O-?-, R6-(CH2)m-NH-?- , or HA375a R1 is or ;

HA375a R2 is , hydrogen, straight or branched chain lower alkyl of up to 5 carbons, or ;

R9 is hydrogen, straight or branched chain lower alkyl of up to 5 carbons, or ;

R3 is lower alkyl of 3 to 5 carbons, -(CH2)n-cyclopentyl, -(CH2)n-cyclohexyl or ;
n is an integer from 1 to 3;
R4 is hydrogen, straight or branched chain lower alkyl of up to 5 carbons, -(CH2)4-NH2, HA375a , or ;

and R5 is straight or branched lower alkyl of up to 5 carbons, , -CH2-(.alpha.-naphthyl), -CCH2-(.beta.-naphthyl), , -CH2-cyclopentyl, HA375a -CH2-cyclohexyl, , or ;

R10 is -(CH2)4-NH2;
R6 and R6' are independently selected from the group consisting of straight or branched chain lower alkyl of up to 5 carbons, cycloalkyl of 4 to 6 carbons, phenyl, 1-naphthyl, and 2-naphthyl;
m and m' are independently selected from the group consisting of zero, one, and two; and HA375a , or .

4. A compound of Claim 3 wherein X is (H3C3)-C-O-?-, , , (H3C)3-C-CH2-?-, (H3C)3-C-NH-?- , (H3C)3-C-?- , , HA375a ,or ;

R1 is ,or ;

R3 is or -CH2CH(CH3)2;

R4 is ; and R5 is benzyl.

5. The compound of Claim 4 wherein:
x is (H3C)3-C-O-?- ;
p is one;
R3 is -CH2CH(CH3)2 ; and R1 is .

6. The compound of Claim 5, N2-[N-[(1,1-dimethylethoxy)carbonyl]-L-phenylalanyl]-N-[(S)-1 [(R)-hydroxy(1H-imidazol-2-yl)methyl]-3-methylbutyl]-L-histidinamide, acetic acid solvate.

HA375a

7. The compound of Claim 4 wherein:

X is (H3C)3-C-O-?- ;
p is one;

R3 is ; and R1 is .

8. The compound of Claim 7, N2-[N-[(1,1-dimethylethoxy)carbonyl]-L-phenylalanyl]-N-[(1S)-2-cyclohexyl-1-[(R)-hydroxy-1H-imidazol-2-ylmethyl]-ethyl]-L-histidinamide, monoacetate salt.

9. The compound of Claim 4 wherein:
x is (H3C)3-C-O-?- ;
p is one;
R3 is -CH2CH(CH3)2; and R1 is .

HA375a

10. The compound of Claim 9, N2-[N-[(1,1-dimethylethoxy)carbonyl] L-phenylalanyl]rN-[(S)-1-[(R)-hydroxy(2-thiazolyl)methyl]-3-methylbutyl]-L-histidinamide, acetate salt.

11. The compound of Claim 4 wherein X is ;

p is one;

R3 is ; and R1 is .

12. The compound of Claim 11, N-[(S)-2-cyclohexyl-1-[(R)-hydroxy(1H-imidazol-2-yl)methyl]-ethyl]-N -[N-(pyrrolidinylcarbonyl)-L-phenylalanyl]-L-histidinamide, dihydrochloride.

13. The compound of Claim 4 wherein X is (H3C)3-C-NH-?-;
p is one;
R3 is ; and R1 is .

HA375a

14. The compound of Claim 13, N-[(S)-2-cyclohexyl-1-[(R)-hydroxy(1H-imidazol-2-yl)-methyl]ethyl]-N2-[N-[[(1,1-dimethylethyl)amino]-carbonyl]-L-phenylalanyl]-L-histidinamide, dihydrochloride.

15. The compound of Claim 4 wherein:

R6 is ;
p is one;
R3 is ; and R1 is .

16. The compound of Claim 15, N-[(S)-2-cyclohexyl-1-[(R)-hydroxy(1H-imidazol-2-yl)methyl]-ethyl]-N -[N-(cyclopentylcarbonyl)-L-phenylalanyl]-L
histidinamide, dihydrochloride.

17. The compound of Claim 4 wherein:

X is (H3C)3-C-CH2-?- ;
p is one;

R3 is ; and R1 is .

HA375a

18. The compound of Claim 17, N-[(S)-2-cyclohexyl-1-[(R)-hydroxy(1H-imidazol-2-yl)-methyl]ethyl]-N2-[N-(3,3-dimethyl-1-oxobutyl)-L-phenylalanyl]-L-histidinamide, dihydrochloride.

19. The compound of Claim 4 wherein:
X is (H3C)3-C-O-?- ;
p is one;

R3 is ; and R1 is .

20. The compound of Claim 19, (1S,2R)-N-[1-(cyclohexylmethyl)-2-hydroxy-2-(2-thiazolyl)-ethyl]-N2-[N-[(1,1-dimethylethoxy)carbonyl]-L-phenylalanyl]-L-histidinamide, 0.5 acetate salt.

21. The compound of Claim 4 wherein:

X is ;

p is one;

R3 is ; and R1 .

HA375a

22. The compound of Claim 21, (1S,2R)-N-[1-(cyclohexylmethyl)-2-hydroxy-2-(1H-imidazol-2-yl)-ethyl]-N2-[N-(4-morpholinylcarbonyl)-L-phenylalanyl]-L-histidinamide, 2.2 trifluoroacetate salt.

23. The compound of Claim 4 wherein:

X is (H3C)3-C-O-?- ;
p is one;
R3 is -CH2CH(CH3)2; and R1 is .

24. The compound of Claim 4 wherein:

X is ;
p is one;
R3 is ; and R1 is .

HA375a

25. The compound of Claim 24, (1S,2R)-N2-[N-[(4-methyl-1-piperazinyl)carbonyl]-L-phenyl-alanyl]-N-[1-(cyclohexylmethyl) 2-hydroxy-2-(1H-imidazol-2-yl)ethyl]-L-histidinamide, trihydrochloride.

26. The compound of Claim 4 wherein X is ;
p is one;
R3 is ; and R1 is .

27. The compound of Claim 26, (1S,2R)-N2-[N-[N2-(cyclobutylcarbonyl)-L-lysyl]-L-phenylalanyl]-N-[1-(cyclohexylmethyl)-2-hydroxy-2-(1H-imidazol-2-yl)ethyl]-L-histidinamide, 3.3 hydrochloride.

28. The compound of Claim 4 wherein:

X is HA375a p is zero;
R3 is ; and R1 is

29. The compound of Claim 28, (1S,2R)-N-[1-(cyclohexylmethyl)-2-hydroxy-2-(1H-imidazol-2-yl)ethyl]-N2-[1-oxo-3-(1-naphthalenyl)-2-(1-naphthalenylmethyl)propyl]-L-histidinamide, dihydrochloride.

30. The compound of Claim 3 wherein:

X is (H3C)3-C-O-?- ;
p is one;
R5 is ;
R4 is ;
R3 is ; and R1 is .

HA375a

31. The compound of Claim 3 wherein:
X is (H3C)3-C-O-?- ;
p is one;

R5 iS ;

R4 is -CH2CH(CH3)2 ;

R3 is ; and R1 is .

32. The compound of Claim 3 wherein:

X is (H3C)3-C-O-?- ;
p is one;

R5 is ;
R4 is hydrogen;

R3 is ; and R1 is .

HA375a

33. The compound of Claim 3 wherein:

X is ;

p is zero;
R4 is ;

R3 is ; and R1 is .

34. The compound of Claim 3 wherein X is (H3C)3-C-? ;
p is one;

R5 is ;

R4 is -(CH2)4 NH2;

R3 is ; and R1 is .

35. A pharmaceutical composition comprising an effective amount of a compound as defined in claims 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33 or 34, or a pharmaceutically acceptable salt thereof, together with a pharmaceutically acceptable carrier therefor.

36. A pharmaceutical composition for use in treating hypertension in a mammalian species comprising an anti-hypertensively effective amount of a compound as defined in claims 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33 or 34, or a pharmaceutically acceptable salt thereof, together with a pharmaceutically acceptable carrier therefor.