AU638093B2

AU638093B2 - Intermediates of peptidylaminodiols

Info

Publication number: AU638093B2
Application number: AU70281/91A
Authority: AU
Inventors: Dale John Kempf; Jay Richard Luly; Jacob John Plattner
Original assignee: Abbott Laboratories
Current assignee: Abbott Laboratories
Priority date: 1985-01-23
Filing date: 1991-02-05
Publication date: 1993-06-17
Anticipated expiration: 2006-01-28
Also published as: AU7028191A

Description

63p 093 S F Ref: 1068101 FORM COMMONWEALTH OF AUSTRALIA PATENTS ACT 1952 COMPLETE SPECIFICATION

(ORIGINAL)

FOR OFFICE USE: Class Int Class 0 S. S Complete Specif.cation Lodged: Accepted: Published: Priority: Related Art: Name and Address of Applicant: Address for Servic.:: Abbott Laboratories 14th Street and Sheridan Road North Chicago Illinois UNITED STATES OF AMERICA Spruson Ferguson, Patent Attorneys Level 33 St Martins Tower, 31 Market Street Sydney, New South Wales, 2000, Australia Complete Specification for the invention entitled: Intermediates of Peptidylaminodiols The following statement is a full description of this invention, including the best method of performing it known to me/us 55* S* 0

S

5845/3 w INTERMEDIATES OF PEPTIDYLAMINODIOLS Technical Field The present invention relates to novel synthetic intermediates employed in the processes for making organic compounds and compositions which inhibit renin. The invention describes these organic compounds and compositions which inhibit renin and a method of treating hypertension with such compounds.

Background Art Renin is a proteolytic enzyme synthesized and stored principally in a specific part of the kidney called the juxtaglomerular apparatus. Any of three different physiologic circumstances may cause the release of renin into the circulation: a decrease in th4 blood pressure entering or within the kidney itself; a decrease in the blood volume in the body; or a fall in the concentration of sodium in the distal tubules of the kidney.

When renin is released into the, blood from the kidney, thL reninangiotensin system is activated, leading to vasoconstriction and conservaticn of sodium, both of which result in increased blood pressure.

The renin acts on a circulating protein, angiotensinogen, to cleave out a fragment called angiotensin I AI TCW/KXW:1052v 2 itself has-only-slight pharmacogic- activity but,-after additional cleavage by a second enzyme, angiotensin converting enzyme (ACE), forms the potent molecule angiotensin II (AlI). The major pharmacological effects of AII are vasoconstriction and stimulation of the adrenal cortex to release aldosterone, a hormone which causes sodium retention. All is cleaved by an aminopeptidase to form angiotensin IIX (AIII), which, compared to All, is a less potent vasoconstrictor but a more potent inducer of aldosterone release.

Inhibitors of renin have been sought as agents for control of hypertension and as diagnostic agents for identification of cases of hypertension due to renin excess.

With these objectives in mind, the renin-anglotension system has been modulated or manipulated, in the past, with ACE inhibitors. However, ACE acts on several substrates other than angiotensin I most notably the kinins which cause such *0 undesirable side effects as pain, "leaky" capillaries, prostaglandin release and a variety of behavioral and neurologic effects. Further, ACE inhibition leads to the accumulation of AI. Although AI has much less vasoconstrictor activity than All, its presence may negate some of the hypotensive effects of the blockade of AII synthesis.

Inhibition of other targets in the renin-angiotensin system such as AII with compounds such as saralasin can block All activity, but would leave unimpaired and perhaps enhance the hypertensive effects of AIII.

On the other hand, there are no known side effects which result when renin is inhibited from acting on its substrate. Considerable research efforts have thus been carried out to develop useful inhibitors of reni. Past research efforts have been directed to renin. Past research efforts have' been directed to -3renin antibodies, pepstatin, phospholipids and substrate analogs such as tetrapeptides and oct peptides to tridecapeptides. These inhibitors either demonstrate po r activity in inhibiting renin production or poor specificity for inhibiting renin only. However, Boger et al. have reported that statine-containing peptides possess potent and specific renin-inhibiting activity (Nature, Vol. 303, p. 81, 1983). In addition, Szelke and co-workers have described polypeptide analogs containing a non-peptide link (Nature, Vol. 299, p. 555, 1982) which also cause potent renin inhibition and show a high specificity for this enzyme.

Disclosure of the Invention According to a first embodiment of the present invention' there is provided a compound of the formula:

OP

2

P

1 NH R'

OP

3 wherein R' is loweralkyl, vinyl or arylalkyl; P 2 and P 3 are independently selected from hydrogen and an 0-protecting group; and P 1 is hydrogen or an N-protecting group; or an acid addition salt thereof.

According to a second embodiment of the present invention there is provided a compound of the formula:

OP

P1NH- CH3 OP3 CH 3 wherein P 1 is hydrogen or an N-protecting group and P2 and P 3 are independently selected from hydrogen and an 0-protecting group; or an acid addition salt thereof.

The specification describes a compound of the formula: 3A u wherein A is hydrogen; loweralkyl; arylalkyl; OR 10 or SR 10 wherein R 10 is hydrogen, lowe~ralkyi or arninoalkyl; NR 11

R

12 wherein R1 and R 12 are independently selected from hydrogen, loweralkyl, aminoalkyl, cyanoalkyl and hydroxyalkyl;

CS

S

S*

S

0O

S.

9 P .855

S

Rl B 0 R 13

B

orS

S

0@SSSS

S

OS@e 9.

5* 0 0S S S 5.5.

S

S..

55 0 S S

S.

4wherein B is NH, alkylamino, S, 0, CH 2 or CHON and R 13 is loweralkyl, cycloalkyl aryl, arylalkyl, alkoxy, alkenyloxy, hydroxyalkoxy, dihydroxyalkoxy, arylalkoxy, arylalkoxyalkyl, amino, alkylamino, dialkylamino, (hydroxyalkyl)(alkyl)amino, aminoalkyl,)'J-protected aminoalkyl, alkylamino-I alkyl, (N-protected)(alIkyl )aminoalkyl, dIralkylauiinoalkyl, (5-membered or 6membered heterocyclic ring or a 5-membered or 6-membered heterocyclic ring to which is fused a benzene ring) alkyl,, or an unsubstituted 5-membered or 6-membered heterocyclic ring or a 5-6imbered or 6-membered heterocyclic ring to which is fused a benzene ring, or a monosubstituted 5-membered or 0i 6-membered heterocyclic ring or a 5-membered or 6-menibered heterocyclic ring to which is fused a benzene ring wherein the substituent is hydroxy, oxo, amino, alkylamino, dialkylamino or loweralkyl, provided that when the or 6-membered heterocyclic ring or the 5-membered or 6-membered heterocyclic ring to which is fused'a benzene ring is satitrated the substituent cannot be oxo; *W is C=O or CHOH; be Ul is CH 2 or NR, provided that when W is CHOH, U is CH; Ris loweralkyl, cycloalkylmethyl, benzyl, 4-methoxybenzyl, halobny,(l-naphthyl)methyl, (2-naphthyl)methyl, (4-imidazolyl)methyl, (x,x-dimethylbenzyl, l-benzyloxyethyl, phenethyl, phenoxy, thiophenoxy or anilino; provided if R1is phenoxy, thiop henoxy or anilino, B is CH 2 or CHOH or A is hydrogen; R 2 is hydrogen or loweraikyl; R 3is loweralkyl, loweralkenyl, EUalkoxy)alkokylalkyl, (thioalkoxy)alkyl, benzyl *0000: or a 5-membered or 6-membered heterocyclic ring or a 5-membered or 6-membered heterocyclic ring to which is fused a benzene ring substituted methyl; R 4 is loweralkyl, cycloalkylmethyl or benzyl; R 5 is vinyl, 0 formyl, hydroxymethyl or hydrogen; R 7 is hydrogen or loweralkyl; R 8 and 7 Rare independently selected from OH and NH 2 and R6is hydrogen, loweralkyl, vinyl or arylalkyl; with the provision that when A is R 13 CONN wherein R 1 is loweralkyl or alkoxy, R 1 is benzyl, l-naphthylmethyl or 2-naphthylmethyl, N is C=O, U is NH, R 3 is loweralkyl or imidazolemethyl, R 4 is benzyl, Ris hydrogen, R 8 is hydroxy and R 9 is hydroxy, then Ris vinyl or arylalkyl when R7is hydrogen and R6is loweralkyl, vinyl or arylalkyl when Ris loweralkyl; and also with the provision that when A is NH 2 or R 13 CONH- wherein R 13 is loweralkyl, alkoxy or benzyloxy, R1'is benzyl, l-naphthylmethyl, 2-naphthylmethyl or (4-imidazolyl)methyl, TCN/KXW: 1052v K) W is C U is NH, R 3 is (4-imidazoilyl)methyl, R 5 is hydrogen, R 6 is hydiogen ,,loweralkyl or arylalkyl, R 7 is hydrogen or loweralkyl, R 8 is hydroxy and R 9 is hydroxy or amino, then R 4 is not loweralkyl; and with the proviso that when A is hydrogen, loweralkyl, arylalkyl,I S hydroxy, amino, alkylamino, (N-protected)amino, or (N-protected)alkylamino; and R 1 is loweralkyl, cycloalkylmethyl, benzyl, 4-methoxybenzyl, (l-naphthyl)methyl, (2-naphthyl)methyl or (4-imidazolyl~methyl; and W is C=O; and U is NR 2 and R 3 is loweralkyl, CH 3 SCH 2 benzyl, (4-imidazolyl)methyl or pyrazolylmethyl; and R8and R 9 are -OH; then- R 5 is other than hydrog en; or pharmaceuticaYly acceptable salts or esters thereof.- In the specification is also described a compound of the formula: 0 R 3

P

tq

II

R 0 R5 R **9 wheei A*sIyakl R1 rS 1 hri 0i yrgn or 0 wherein A is NH, alkyl; OR 10 or SR 0 HereinHan R1 is hyd oe ly amno (hy5 loweralkylaloramino NR 1

R

2 whrin R 11 nd R 1 a-roee mnakl independental (Nel ectedm hyrogen, oealkyl, aialkyl, cynoaylky an hyroylkyl;gtowihisfsdabezn in)akl TCW/KXW: 1052v -6- 6 O NL 0 2 OH

OOH

0 OH

OH

SN

The term "N-protecting group" or "N-protected" as used herein refers to those groupj intended to protect nitrogen atoms against undesirable reactions during synthetic procedures or to prevent the attack of exopeptidases on the final compounds or to increase the solubility of the final compounds and includes but is not limited to acyl, acetyl, pivaloyl, t-butylacetyl, t-butoxycarbonyl(Boc), benzyloxycarbonyl (Cbz) or benzoyl groups or an L- or 0- aminoacyl residue, which may itself be N-protected S similarly.

The term "loweralkyl" as used herein refers to straight or branched chain alkyl radicals containing from 1 to 6 carbon atoms including but not limited to methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, secbutyl, n-pentyl, 1-methylbutyl, 2,2-dimethylbutyl, 2-methylpentyl, 2,2dimethylpropyl, n-hexyl and the like.

The term "loweralkenyl" as used herein refers to a loweralkyl radical S which contains at least one carbon-carbon double bond.

The term "arylalkyl" as used herein refers to an unsubstituted or substituted aromatic ring appended to an alkyl radical including but not limited to benzyl, 1- and 2-naphthylmethyl, halobenzyl and alkoxybenzyl.

The term "aminoalkyl" as used herein refers to -NH 2 appended to a loweralkyl radical.

to The term "cyanoalkyl" as used herein refers to -CN appended to a loweralkyl radical.

The term "hydroxyalkyl" as used herein refers to -OH appended to a loweralkyl radical.

The term "alkylamino" as used herein refers to a loweralkyl radical appended to an NH radical.

The term "cycloalkyl" as used herein refers to an aliphatic ring TCW/KXW:1052v R I i i Sit 61 1 I 1 T 7 -7having 4 to 7 carbon atoms.

The term "cycloalkylmihyl" as used herein refers to a cycloalkyl group appended to a methyl radical, including but not limited to cyclohexylmethyl.

The term "aryll' as used herein refers to a substituted or, unsubstituted aromatic ring including but not limited to phenyl,'naphthyl, halophenyl and alkoxyphenyl.

The terms "alkoxy" and thioalkoxy" as used herein refer to R140and R I 4 respectively, wherein R14 is a lowera,1;yl group.

The term "alkenyloxy" as used herein refeSto R 15 0- wherein R5 ist 15 weri is an unsaturated alkyl group.

The term "hydroxyalkoxy" as used herein refers t6 -OH appended to an alkoxy 'Ijdical.

The term "dihydroxyalkoxy" as used herein refers to an alkoxy radical which is disubstituted- with -OH radicals.

The term "arylalkoxy" as used herein refers to an aryl appended to an .0 alkoxy 0: The term "arylalkoxyalkyl" as used herein refers to an arylalkoxy appended to a loweralkyl radical.

20 The term "(thioalkoxy)alkyl" as used"herein refers to thioalkoxy appended to a loweralkyl radical.

The term "dialkylamino" as used herein refers to,-NR 16

R

17 wherein

R

16 and R17 are indepe;,ently selected from loweralkyl groups.

The term "E(alkoxy)alkoxylalkyl" refers to an alkoxy group appended 2o to an alkoxy group which is appended to a loweralkyl radical.

00 The term "(hydroxyalkyi)(alkyl)amino" as used herein refers to

-NR

18

R

19 wherein R 18 is hydroxyalkyl and R 1 is loweralkyl.

"The term "N-protected aminoalkyl" as used herein refers to which is appended to a loweralkyl group, wherein R20 is an N-protecting group.

The term "alkylamn\)alkyl" as used herein refers to NHR 21 appended to a loweralkyl radical, wherein R21 is a loweralkyl group.

The term "(N-prclected)(alkyl)aminoalkyl" as used herein refers to NR20R21 which is appended to a loweralkyl radical, wherein R20 and R21'are as defined above.

The term "dialkylaminoalkyl" as used herein refers to NR 22

R

23 is appended to a loweralkyl radical wherein R22 and R 23 are independently TCW/KXW:1052v -8sselected from loweralkyl.

The term "(heterocyclic)alkyl" as used herein refers to a heterocyclic group appended to a loweralkyl radical, including but not limited to imidazolylalkyl.

The term "O-protecting group" as used herein refers to a substituent which protects hydroxyl groups and includes but is not limited to substituted methyl ethers, for example, methoxymethyl, benzyloxymehyl, 2-methoxyethoxymethy1, 2-(trimethylsilyl)ethoxymethyl and tetrahydropyranyl; substituted ethyl ethers, for example, 2,2,2-trichloroethyl, t-butyl, benzyl and triphenylmethyl; silyl ethers, for example, trimethylsilyl, t-butyldimethylsilyl and t-butyldiphenylsilyl; cyclic acetals and ketals, for example, methylene acetal, acetonide and benzylidene acetal; cyclic ortho esters, for example, methoxymethylene; cyclic carbonates; and cyclic boronates.

The term "heterocyclih7 ring" or "heterocyclic" as used herein refers 0 to a 5-membered or 6-membered heterocylic ring or a 5-membered or 6-membered heterocyclic ring to which is fused a benzene ring. The "heterocyclic ring" or "heterocyclic" generally contains from one to three .heteroatoms selected from the group consisting of nitrogen, oxygen, and .0 sulfur; having various degrees of unsaturation; wherein the nitrogen and S sulfur heteroatoms may optionally be oxidized; wherein the nitrogen heteroatom may optionally be quarternized; and including any bicyciic group in which any of the above heterocylic rings is fused to a benzene ring.

Heterocyclics in which nitrogen is the heteroatom are preferred. Fully saturated heterocyclics are also preferred. Preferred heterocyclics are: pyrryl, pyrrolinyl, pyrrolidinyl, pyrazolyl, pyrazolinyl, pyrazolidinyl, imidazolyl, imidazolinyl, imidazolidinyl, pyridyl, piperidinyl, p-rkzinyl, piperazinyl, pyrimidinyl, pyridazinyl, oxazolyl, oxazolidinyl, thoh4zoly1, isoxazolidinyl, morpholinyl, thiazolyl, thiazolidinyl, isothiazolyl, 30 isothiazolidinyl, indolyl, quinolinyl, isoquinolinyl, benzimidazolyl, benzothiazolyl, benzoxazolyl, furyl., thienyl and benzothienyl.

Saturated heterocyclics may be .unsubstituted or monosubstituted with hydroxy, oxo, amino, alkylamino, dialkylamino or loweralkyl. Unsaturated heterocyclics may be unsubstituted or monosubstituted with hydroxy, amino, alkylamino, dialkylamino or loweralkyl.

The most preferred heterocylics are as follows: TCW/KXW:1052v 9- N

N

O 0

OH

I

N

/ND

(CH, )n

X

wherein n is 1 or 2 and X is N, NH, 0, S, provided that X is the point of connection only when X is N,

N

wherein is NH, N-oweraky, S, or SO or wherein Y is NH, N-loweralkyl, 0, S, or SO or t US)S*e a he 4 4 hr44 a 4

Z,

t-

ZI

2f wherein Z1 is N, 0, or S and not the point of connection and Z 2 is N when it is the point of connection and NH, 0 or S when it is not the point of connection.

'b The terms "Ala", "His", "Leu", "Phs", "Tyr", "Cys", "Gly", "Lys", S"Sar" and "Pro" as used herein refer to alaniieg, histidine, leucine, S phenylalanine, tyrosine, cysteine, glycine, lysine, sarcosine and proline, r'especti vely.

Most of these compounds may be made as shown in Scheme I. The amino '1 "15 diol intermediate of the invention 5 represents a transition-state mimic for the Leu-Val scissile bond of the renin substrate, angiotensinogen.

Incorporation of this amine into the angiotensinogen sequence in place of Leu-Val Ile-Protein provides potent inhibitors of human renin. For example, acylation of amine 5 with an acyl-Phe-His-OH residue or other appropriately modified amino acid derivatives produces small peptide analogues which are potent renin inhibitors.

TCK KXW:1 052v 4 a. 0 0 0 0* 0903 a *0 9 0 t 0 490 0 C L) 0 I ~4L06

P

1

-NII

R

A-M.

2 PEPTIDE-NIIn R it 6 I~x

P

1 I is an N-protecting group; P 2 and P 3 are independently selected from hydrogen or an 0-protecting group.

R is loweralkyl, cycloalkylmethyl or benzy].

R' is hydrogen, loweralkyl, vinyl or arylalkyl.

lIX is an acid.

35-2528i~ 11 More particularly, the process shown in Scheme I describes an N-protected-aminoaldehyde 1 (P 1 is an N-protecting group) which is treated with an yield to give the corresponding allylic amine 2. Oxidation gives a diol of the invention 3 (P 2 and P 3 are both hydrogen), N-deprotection gives 4; and free-basing gives amine 5. Either intermediate A or 5 can be converted to 7 by standard peptide coupling methods. The same sequence (3 7) can be carried out with hydroxy protecting groups present (where P 2 and/or P 3 are 0-protecting groups), the final step then being 0-deprotection. Alternatively, allylic amine 2 may be N-protected, peptide coupled using standard methods to give 6, and then oxidized to give the desired peptide diols I.

The protected aminodiol fragment of the invention may be alternatively prepared as shown in Scheme II. Aldehyde 3 (prepared, for example, by oxidation of alcohol 8) is converted to its cyanohydrin Addition of an organometallic reagent (such as a Grignard reagent) and acidic workup provides ketone 12. Reduction of ketone 12 then provides the desired protected aminodiol 13.

05* se 'j TCW/KXN:1052v

S

S S 55 S S 0* 5 0 0.S *5 ese S S S S S S S S S S S S S S C S S S S C S S. 055 5 555 SCHEME 11 Pj-N~lOXIDATION ofIl 0 REDUCTIO14 R' iI N-hMgX pl -R and R' are as defined for Scheme 1, excapt chat R' cannoc be hydrogen.

13 The following Examples will serve to further illustrate preparation of the intermediates of the invention and the final compounds.

Example 1 2(S)-t-Butyloxycarbonylamino-l-cyclohexylbut-3-ene A 0 0 C solution of potassium hexamethyldisilazide (22.9 mmol in 115 mL of 5:1, tetrahydrofuran (THF): dimethyl sulfoxide (DMSO) was added dropwise to triphenylmethylphosphonium iodide (24.81 mmol). After stirring at 0°C for 1 hour, the solution was cooled to -78 0 C and a solution of Boc-cyclohexylalaninal [4.90 g, 19.08 mmol, prepared by Swern oxidation (Mancuso, Huang, and Swern, J. Org. Chem. 1978, 43, 2480) of Boccyclohexylalaninol] in dry THF (95 mL) was added. After stirring at -78°C for 1 hour, the mixture was allowed to warm to room temperature. The reaction mixture was quenched with aqueous ammonium chloride and extracted S with ether (2x300 The combined organic phase was washed with 10% HC1 15 (200 mL), saturated NaHSO3 (2x200 mL), H 2 0 (2x200 mL), saturated NaHCO3 (2x200 mL), and brine (200 mL), dried (MgSO 4 filtered, and 3 evaporated. The residue was purified by chromatography (40 m S'02; S ether: hexane, 15:85) to give the desired compound in 60% yield. Mass spectrum: 254.

Example 2 Boc-Phe-Ala Amide of (2S)-Amino-l-cyclohexylbut-3-ene The resultant compound of Example 1 (310 mg, 1.22 mmol) was dissolved in 1 M anhydrous HC1 in anhydrous methanol (35 mL). After 22 hours, the solvent was evaporated to give 230 mg of the corresponding amine 5 hydrochloride which was used without further purification.

TCWIKXO: 1052v To a stirred -13 0 C, solution of Boc-Phe-Ala-(408' mg, 1.21 mmo 1) in dry THF (41 mt) containing N-methylmorpholine (122 mg, 1.21 mmol) was added isobutyl chloroformate (165 mg, 1.21 mmol) dropwise.

After 3 minutes, a -13"C solution of the above amine hydrochloride (230 mg, 1.21 mmol) in 1:1, THF:dimethyl for amide (DMF) (4 mL) containing N-methylmorpholine (122 mg) was added dropwise. The mixture was warmed to room temperature for 2 hours. Evaporation provided a residue wbhch was partitioned between ethyl acetate ji mL) and 1 M H 3

PO

4 (10 mL) The organic phase was washed wiLh brine (10 mL), saturated NaHCO 3 (10 mL), and brine (10 mLt). Drying, filtering, evaporating, and chromatographing (55 g SiO 2 95':5, CH 2

C

12

:CH

3

OH)

ir gave the desired compound (462 mg, 81%).

0 *see*.

a S.

.00.

*5 Si

S..

Example 3 Boc-Phe-Ala Amnide of 3(S)-Amino-4-cyclohery1- 1,2(R,S)-dihydroxybutane S 56 S 5 5* S. C S

S.

S

To a stirred solution cE the resultant compound of Example 2 (100 mg, 0.212 mmol) in THF (5 mL) were added Os0 4 solution (0.065 mL of a 2.5 W/V% solution in t-butanol) and N-methylmorpholine N-oxide (57 mg, 0.424 mmol) sequentially. After 4.5 hours, brine 25 mLt) was added, and the mixture was extracted with ether S(4x8 mLt). The combined organic phase was washed with 10% Na 2

SO

3 (3 x 6 mL), 0.1 M H 3

PO

4 (5 mt), and brine (5 mL). Drying, filtering, and evaporating provided the desired product (97 mg, Mass spectrum: M= 505.

Example 4 3(S)-t-Butyloxycarbonylamino-4-cyclohexvl- 1,2(R,S)-dihydroxvbutane To a stirred solution of 2(S)-t-butyloxycarbonylamino-l-cycl ohexylbut-3-ene (1.00 g, 3.95 mmol) in THF (20 mL) were added OsO 4 solution (1.2 mL of a W/V% solution in t-butanol) and N-methylmorpholine N-oxide (1.07 g, 7.90 mmol). After 24 hours, the mixture was partitioned between ether (50 mL) and brine (25 mL). The layers were separated, and the organic phase was extracted with ether (3 x 25 mL). The combined organic phase was washed with 10% Na 2

SO

3 (4x10 mL), 1.0 M H 3

PO

4 (2x8 mL), and brine (15 mL).

Drying and evaporating provided the desired product as an oil (1.14 g, 100%). 'H NMR shows a 1:1 mixture of diastereomers (NH 4.43 and 4.56 ppm).

Example Boc-Phe-His Amides of 3(S)-Amino-4-cyclohexyl- 2(R, S)-hydroxy-l-t-butyldimethylsilyloxybutane The resultant compound of Example 4 (1.10 g, 3.82 mmol) was treated with anhydrous 1M HC1/CH 3 0H mL) for 16 hours at which time evaporation and drying provided the corresponding amine hydrochloride (0.85 g, 100%).

9 *9 9 ei To a suspension of the above hydrochloride salt (344 mg, 1.54 mmol) and imidazole (105 in dichloromethane (15 mL) were added triethylamine (156 mg) and t-butyldimethylsilyl chloride (232 mg). The 25 solvent was evaporated after 31 hours, and the residue was then re-dissolved in anhydrous dimethylformarmide (DMF, 15 mL) Boc-Phe-His (619 mg) and l-hydroxybenzotriazole (HOBT, 312 mg) were then added. After cooling the stirred solution to -23°C, 1,3-dicyclohexylcarbodiimide (DCC, 318 mg) was added. The mixture was warmed to room temperature 3 hours later. After 13 hours the solvent was evaporated in vacuo, and the residue was dissolved in ethyl acetate (40 mL), filtered, washed with saturated NaHCO 3 (2x10 mL) and brine (10 mL), and dried (Na 2

SO

4 Filtration and evaporation provided a residue which was chromatographed 1 i 16 on silica gel eluting with dichloromethane/methanol mixtures to give 441 mg of the desired product.

Mass spectrum: 686.

Anal. calcd. for C 36 H 59

N

5 Si: C, 63.0; H, 8.7; N, 10.2. Found: C, 62.8; H, 9.0; N, 9.9.

Example 6 Boc-Phe-His Amides of 3(S)-Amino-4-cyclohexyl- 1,2(R)-dihydroxybutane To a stirred solution of the resultant product of Example 5 (200 mg, 0.291 mmol) in anhydrous THF mL) at 0 C was added tetrabutylammonium fluoride (0.58 mL of a 1 M solution in THF). The solution was warmed 15 to room temperature for 4 hours and then evaporated.

The residue was dissolved in chloroform and washed with water (3X) and brine Drying and evaporating provided a gum which was treated with hot ethyl acetate (8 mL). Cooling and filtration provided 25 mg of the desired material. Mass spectrum: 572.

Anal. Calcd for C 30

H

45

N

5 0 6 1/2H 2 0: C, 62.1; H, 8.0; N, 12.1. Fotnd: C, 62.4; H, 8.2; N, 12.0.

0g*S Example 7 25 (4S)-2,8-Dimethyl-4-[(toluenesulfonyl)amino]- To a stirred -78°C solution of toluenesulfonyl (Ts)-Leu (15 g, 53 mmol) in dry THF (240 mL) was added 00 4 butyl lithium (57.8 mL of a 0.91 M solution in hexane) followed 15 minutes later by isopentyl magnesium bromide (185 mL of a 0.8 M solution in THF). The mixture was heated at reflux for 3 days, then cooled and poured into 0°C 1 M HC1 (500 mL). The layers were separated and the aqueous phase was extracted with ether (3x150 mL). The combined organic layers were washed with saturated NaHCO 3 (2x150 mL) and brine (150 mL). Drying and evaporating provided a residue which was chromatographed on silica gel to give 7.43 g of the desired product. Mass spectrum: 340.

Anal. calcd. for C 18

H

29 NO S: C, 63.7; H, 8.6; N, 4.1. Found: C, 64.0; H, 8.6; N, 4.1.

Example 8 (4S)-2,8-Dimethyl-5-hydroxy-4-[(toluenesulfonyl) To a stirred 0°C solution of the resultant compound of Example 7 (79 mg, 0.23 mmol) in dry THF.

(8 mL) was added vinyl magnesium bromide (1.5 mL of a 1.0 M solution in THF) dropwise. The mixture was warmed (room temperature, 10 hours), quenched (8 mL H20 2 mL brine), acidified with 0. 1 M H 3

PO

4 and extracted with ether (3 x 4 mL). The combined ether phase was washed (4 mL brine), dried (Na 2

SO

4 filtered, and evaporated to give 81 mg of the desired product as a 4:1 mixture of diastereomers.

Example 9 Boc-Phe-Ala Amide of (4S)-Amrino-2,8-dimethyl- S* To a solution of the resultant compound of 25 Example 8 (400 mg, 1.09 mmol) in liquid ammonia (80 mL) was added sodium (150 mg, 6.5 mmol). After 6 hours the ammonia was allowed to slowly evaporate under a stream S. of nitrogen. Benzene (50 mL) and 1:i, ethanol:water s'e 3 (20 mL) were added with stirring. The layers were separated, and the,, aqueous phase was extracted with ether. The combined organic phase was dried (Na 2

SO

4 filtered, and evaporated to give 85 mg of the desired product.

Following the procedure of Example 2, but replacing the amine hydrochloride and N-methylmorpholine with the above resultant product, gave the desired major diastereomer in 35% yield after chromatography. FAB mass spectrumn: -570.

Anal. calcd. for C 0

H

49 N 0 5 C; 67,8;I H, 9.3; N, 7.9. Found: C, 67.7; H, 9.6; N, 7.3.

Example Boc-Phe-Ala Amnide of (3S)-Amino-2-hydroxy-2i Following the procedure of Example 3 with the resultant compound of Example 9 except replacing N-methylmorpholine N-oxide with aqueous' Na1O 4 gave the.

desired compound.

ExaMple 11 Boc-Phe-Ala Amide of 3-Amino-1,2-dihydroxy- Treatment of the resultant compound of PExample 10 with one equivalent of NaBH 4 in methanol provided the desired compouind after aqueous work-up.

Example 12 Bo-Phe-Ala Arnide. of 3-Amino-i, 2-dih droxy 2-i Scale up of the procedure of Example 8 led to :the isolation of the,. minor diastereomer pure after chromatography, Treatment as in Examples 9, 10, and 11 p, ,ovided-the desired isomer of the resultant product of "goo Example 11.

Examvle 13 j 2(S)-t-Butyloxcycarbonylarnino-1-cyclohexyl- 6,-methylhept-3-ene To a stirred -8C solution of Boc-cyclohexylalanine methyl ester (40 g, 140 mmcl) in anhydrous toluene (250 m.L) was added di isobutyl aluminum hydride (130 1.5 14 solution in toluene, 121.4 mL) at 19 a rate to keep the internal temperature below -60 0

C.

After stirring for an additional 20 minutes at -78 0

C,

the aldehyde solution is used immediately as described below.

To a potassium hydride (35% dispersion in oil, 32.09 g) suspension in a o0C mixture of anhydrous THF/DMSO (1000 mL/200 mL) under dry N 2 was added l1,1,,3,3,3-hexamethyldisilazane (209 49.07 g) dropwise. After stirring at 0°C for 1 hour, the resulting solution was added via cannula to a 0 C flask containing isopentyltriphenylphosphonium bromide (209 125.66 .The mixture was stirred vigorously for 1 hour at which time it was cooled to -78°C. The -78°C 0 aldehyde solution prepared above 'was then added v\ 15 cannula. After stirring at -78°C for 15 minutes, the *0 mixture was allowed to slowly warm to rocm temperature and then heated to 40°C for 12 hours. The mixture was then cooled to' room temperature and quenched with Smethanol (7.65 mL) followed by aqueous Rochelle salts (100 mL saturated solution and 500 mL H 2 The mixture was then extracted with ethyl acetate The combined extracts were washed with water and brine.

Drying (?igSO 4 and evaporating provided crude alkene which was chromatographed on silica gel (ether/hexane) 25 to give 16.5 g of the desired compound as an 85:1,5 mixture of cis:trans isomers. Mp=53-55 0 C. Mass spectrum: M4 =309.

Anal. calcd. for C 9

H

35 NO2: C, 73.7; H, 11.4; N, 4.5. Found: C, 73.8; H, 11.4; N, Example 14 2(S)-t-Butyloxycarbonylamino-l-cyclohexyl-3,4dihyd.roxy-6-methylheptane: The and 3(S)4(R) Diastereomers To a solution of the resultant compound of Example 13 (8.50, 27.5 mmol) in dry THF (150 mL) were added 0s0 4 (2.8 mL of a 2.5% solution in t-butanol and N-methylmorpholine N-oxide (9.28 g, 68.7 mmol). After 4 days the mixture was partitioned between ether (200 mL) andX brine (100 mL). The aqueous layer was backextracted with ether (2x100 mL), and the combined organic phase was washed with 10% Na 2 50 3 0.1 M4 H POg and brine. Drying (MgSO 4 and 'evaporating provided a residue (10.81 g) which was chromatographed on silica gel to elute a 60% yield of the 4 diols in the following order.

Mass spectrum: Anal. calcd. for C 19

H

37 NO 4 C, 66.4; H, 10.9; N, 4.1. Found: C, 66.4; H, 10.8; N, 3.9.

cac. Mass spectrum: 344. 'Anal.

bac f or C 19

H

37 N0 4 C, 66.4; H, 10.9; N, 5.1.

Found: C, 66.4; H, 11.1; N, 6S~e Mass spectrum: (14-iH) =344.

Mass spectrum: =344. Anal.

calcd. f or C1 9 H 37 NO 4 C, 66,~ H, 10.9; N, 4.1.

Found: C, 66.0; H,,10.7; N, Examole *Boc-Phe-His Amide of 2(S)-Am ino-1-cyclohexyl- 3(R).4 (S )-dihyrdroxy-6-nt ethylheptane 5The 3 ,4 dia'stereomer of Example 14 was deprotected with HCl/methanol, and the resulting product was coupled to Boc-Phe-His .using l-hydroxybenzo- *~.triazole and l,3-dicyclohexylcarbodiimide according to the procedure of Example 5. The desired product was obtained in 40-60% yield. Mass spectrum: 628.

Anal.", calcd. for C H N' 0 H 0: C 34 535 6 2 63.2; H, 10.8. Found: C,'63.2; H, 8.4; N, 10.5.

Example 16 3oc-Phe-H is Amnide of- 2(S)-Amino-1-cyclohexyl- 3(S) ,4(S)-dihydroxy--6-methylheptane-, Following the pr~ocedure of Example 15, but replacing the diastereomer with the 3(S 4 (S) diastereoner gave the desired compound. Mass spe%';,rum: 628. 1 Anal. calcd." for C H N 0 6 1/2H 0: C, 64. 1; H, 8. 6; N, 11.0. Found: C, 64. 0; H. 8. 6; N, Example 17 Boc-Phe-His Amide of 2(§S)-Amino-l-cyclohe'vl- 4(R)-dihydrox, -6-methylheptane Following the procedure of Example 15, but .replacing the, diastereomer with the 3(R),4(R) *diastereomier gave the desired compound. M ass spectrum: AnHal 28. calcd. for C 34

H

53

N

5 0 6 11 2

C,

63.2; H, 8.6; N, 10.8. Found:,C, 63)1; H, 8.5; N, 10.7.

-Exarnole 18 Boc-Phe-His Amide of 2(S)-AminOj-l-c'cIohexyl- 4(R)-dihydroxy-6-methylheptane 25 Following the procedure of Example 15, but replacing the diastereomer IWIIth thl--i 3(S),4(R) diastereomer gave the desired compound. Mass spectrum:' (M-iH) 628.

Anal. calcd. for C 34

H

53 7 5 0 6 3/4H 2 0: 730 C, 63. 7; H, 8. 6; N, Found: C, 63.8; H, 8.8; N, 1- Example 19 A. 3(R,S)-hydroxy-l-pentene To a stirred -78°C solution of Boc-cyclohexylalanine methyl ester (10.2 g, 35.8 mmol) in dry toluene (60 mL) was added diisobutylaluminum hydride (34 mL of a 1.5 M solution in toluene). After minutes, vinyl magnesium bromide (108 mL of 1 M solution in THE) was added. After stirring for 15 hours at 0°C, the mixture was carefully quenched with methanol, treated with Rochelle salts (22 mL of saturated aqueous solution in 140 mL H 2 and filtered. After extracting the solids 5 times with aethyl acetate, the extracts and filtrate were combined 15 and the organic phase was washed with brine, dried, filtered, and evaporated to an oil (10.2 g), a Chromatography on silica gel eluting with hexane/ethyl acetate mixtures provided 6.1 g of the desired product.

Anal. calcd. for C 16

H

29

NO

3 1/4H 2 0: C, 66.8; H, 10.3; N, 4.9. Found: C, 66.9; H, 10.2; N, 4.7.

B. 4(S)-Cyclohexylmethyl-5(R,S)-vinyl-2- S" oxazolidinone The resultant product of Example 19A (2.80 g, *1 9.88 mmol) in dry dimethylformamide (DMF) (50 mL) was Sadded to a stirred suspension of NaH (593 mg of a dispersion in oil, 14.8 mmol, hexane washed) in dry DMF mL). After 3 hours, the mixture was quenched (750 mL water 100 mL brine) and extracted with ether (5x100 mL). The combined organic phase was washed with brine (3x50. mL), dried (MgSO 4 filtered, and evaporated to an oil 2.23 g. The NMR spectrum of the crude product revealed an 82:18 mixture of 5 S:5 R diastereomers.

Silica gel chromatography gave 80% recovery of pure diastereomers. 5 S: Anal. calcd. for C 12

H

19

NO

2 C, 68.9; H, 9.1; N, 6.7. Fou 68.4; H, 9.2; N, Mass spectrum: 210. 5 R: Mass\ spectrum: 210.

C. 5(R)-Carboxy-4(S)-cyclohexylmethyl-2-oxazolidinone To a solution of the compound from Example 19B (1 g, 4.78 mmol) dissolved in 16 mL of benzene and 3 mL of acetic acid was added a solution of 3.01 g of potassium permanganate in 16 mL of water. The resultant two-phase mixture was vigorously stirred and treated by portionwise addition with 153 mg of tetrabutylammonium bromide. After stirring for 2 hours at room temperature, the mixture was quenched with aqueous sodium bisulfite, acidified to pH=3, and extracted with .ow ethyl acetate. Drying and evaporating gave the desired product as an oil in 59% yield.

D. 4(S)-Cyclohexylmethyl-5(R)-[3-(3hydroxvDentvl)1-2-oxazolidinone 0 0

I

0.0 To a solution of the compound from Example 19C dissolved in tetrahydrofuran and cooled to -78 0 C was added 3.5 equivalents of ethyl magnesium bromide. After stirring at -78 0 C for 1.5 hours and at room temperature for 1 hour, the reaction mixture was quenched with water and extracted with ether. The dried ethereal extract was evaporated to afford a 73% yield of product.

E. 2(S)-Amino-l-cyclohexyl-3(R)-3,4dihydroxy-4-ethylhexane A solution of the compound from Example 19D (1.69 mmol) and barium hydroxide octahydrate (3.38 mmol) in dioxane (60 mL) and water (40 mL) was heated at reflux under N 2 for 21 hoirs. The solid barium carbonate was filtered and the filtrate was partially evaporated. The residue was diluted with water and the resulting solution was extracted with ether. The.

organic extract was washed with brine solution, dried over MqSO 4 and evaporated to give the desired product in 76% yield.

F. Boc-Phe-His Amide of 2(S)-Amino-l-cyclohexyl- 3(R)-3 ,4-dihydroxy-4-ethylhexane The resultant product of Example 19E was' coupled to Doc-Phe-His using l-hydroxybenzotriazole and l,3-dicyclohexylcarbodiimide according to the procedure of Example 5 to give the desired pr oduct in 55% yield.

Example Boc-His Amnide of 2(S)-Amino-l-cyclohexyl- 3(R),4(S)-dihydroxy-6-rnethylheptane The procedure of Example 15 was followed except Boc-Phe-His was replaced with Boc-His. Mass spectrum: 480.

calcd. for C 25 H N 0 3/4H 0: 20 25 444 52 C, 60.8S; H, 9. 1; N, 11..3. Found: C, 60.9; 9.2; N; Example 21 ~:'.TBA-CHA-His Amnide of 2(S)-Amino-l-cyclohexyl- 25 3(R) ,4(S)-dihydroxy-6-rnethylhep~tane.

resultant compound of Example 20 was deprotected with HCl/methanol, and the resulting pro~duct was coupled to t-butylacetyl-cyclohexylalanine (THA- CHA) using the DCC/HOBT method of Example 5. HRM'S calcd. for C 35

H

61 N 5 0 5 P 632.4751. Found: 632.4759.

Example 22 Ethoxycarbonyl-(OCH 3 )Tyr-His Amide of 2(S)- Amnino-l-cyclohex).-3 ,4(Sg)-dihydroxy-6--methylheptane Using the procedure of Example 21, but replacing TBA-CHA with ethoxycarbonyl-(OCH3)Tyr-His gave the desired compound. Mass spectrum: (M+iH) 630.

Example 23 Acetyl-N-methylPhe-His Amide of 2(S)-Amiino-l-.

cyclohexyl-3 ,4(S)-dihydroxy-6-methylheptane Using the procedure of Example 21, but replacing TBA-CHA with acetyl-N-methylPhe gave the desired compound. Mass spectrum: M+ 583.

Example 24 15 2. Ac-Pi-His Amnide of 2(S)-Amino-l-cyclohexyl- 3(R) ,4(S)-dihydroxy-6-methylheptane *Using the procedure of Exaznpls 21, b'it replacing TBA-CHA with O-acetyl-L-3-phenyllactic acid (Ac-Pl-OH) gave the desired compound. HRM4S calcd. for C 31

H

46 N 4 0 6 (Z4+H) 571.3495. Found: 571.3489.

Example see#$:.

P1-His Amide of 2(S)-Amino-l-cyclohexyl-3(R) dihydroxy-6-methylhept ane The resultant compound of Example 24 (37.4 mg, *0.065 mmol) in MeOH at 0 0 C was treated, with K 2 C0 3 (9.1 mg, 0.065 mmol) for 30 minutes at 0 0 C. Evaporation provided, a residue which was partitioned between ethyl acetate and water. The organic phase was washed (brine), dried (MgSO 4 and evaporated to give the desired compound (32 mg, Mass spectrum: =529.

Anal. calcd. for C 31

H

46 N 4 0 6 1f/2H 2 0: 64.8; H, 8.4; N, 10.4. Found: C, 64.6; H, 8.3; N, 10.1.

ExamPle 26 Boc-l-Nal-His Amide of 2(S)-Amino-1-cyclohexyl- 3(R),4(S)-dihydroxy-6-methyl-heptane Using the procedure of Example 21, but replacing TBA-CHA with Boc-l-naphthylalanine (Boc-1-Nal) provided the desired compound. Mass spectrum: =678.

Example 27 Dba-His Amide of 2(S)-Amino-l-cyclohexyl- 3(R),4(S)-dihydroxy-6-methylheptane Using the procedure of Example 21, but e replacing TBA-CHA with 2,2-dibenzylacetic acid (Dba-OH) 15 gav- the desired compound. HRIMS calcd. for C C 3 6

H

5 0 N 0 4 603.3910. Found: 603.3899.

Example 28 0 goe Pp-His Amide of 2(S)-Amino-1-cyclohexyl-3(R) dihydroxy-6-methylheptane Using the procedure of .Example 21, but replacing TBA-CHA with 3-phenyl-propionic acid .(Pp-OH) gave the desired compound. Mass spectrum: C g~ 513.

Anal. calcd. for C 2 9

H

44

N

4 0 4 1/2 H 2 0: C, 66.8; H, N, 10.7. Found: C, 66.6; H, 8.8; N, 10.5.

Example 29 Ethoxycarbonyl-Phe-His Amide of 2(S)-Amino-lcyclohexyl-3(R),4(S)-dihydroxy-6-ethylhelptane Using the procedure' of Example 21, but replacing TBA-CHA with ethoxycarbonyl-Phe gave the desired product. Mass spectrum: 600.

Anal. calcd. for C 3 2 H49N506 1/2H 0: 63.1; H, 8.3; N, 11.5. Found: C, 62.8,; H, 8. 3; N, 11.4 Example Ac-Phe-His Amide of 2(S)-Amrino-l-cyclohexyl- 3(R) ,4(S)-dihydroxy-6-methylheptane Using the pitocedure of Example 21, but repl &cing TBA-CHA with acetyl(Ac)-Phe gave the desired produ ct. Mass spectrum: 570.

calcd. for C 31 H 47N 50 5 l/2H 2 0: C, 64.3; H, 8.2; N, 12. 1. Found: C, 64.2; H, 8.3; N, 12.0.

Example 31 Boc-Leu-His Amide of 2(S)-Amino-l-cyclohexyl- -5 4(S)-dihydroxy-6-methylheptane *Using the procedure of Example 21, but replacing TBA-CHA with Boc-Leu gave thie desired product. Mass spectrum: =594.

Anal. calcd. for C 31

H

55 N 5 0 6 1/2H 2 0: C, 61.8; H, 9.4; N, 11.6. Found: C, 61.8; H, 9.3; N, 11.6.

Example 32 Tbac-Phe-His Amide of 2(S)-Amino-7.-cyclohexvl- 4(S)-dilhydroxy-6-methylheptane .Using the procedure of Example 21, but replacing TiA-CHA with t-butyl-aminocarbonyl-Phe :*(Tbac-Phe) Ngave the desired product. Exact mass calcd for C 34

H

55 N6 05:; 627.4233. Found: 627.4226.

Examnle 33 Hoc-Phe-Ala-Amide-of 2(S)-Amino-l-cyclohexyl- 3(R) ,4(S)-dihydroxv-6-methylheptane Using the procedure of 2, but replacing the resultant compound of Example 1 withl the 3(R),4(S) 28 diastereomer of Examplie 14 gave the desired compound.

Mass spectrum: 560 Anal. callcd,;, for C 3 1

H

1 3 0 6 C, 66.3; H, 9.1; N, 7.5. Found,' C, 66.0;"H, 9.2; N, 7.3.

.Example 34 Boc-Phe-Phe Amide of 2(S)-Amino-l-cyclohexyLj 3(R),4(S)-dihydroxy-6-methylhentane Using the procedure of Example 33, but replacing Boc-Phe-Ala with Boc-Phe-Phe, gave the desired produc t. Mass spectrum: 638.

Anal. calcd. for C 3 7 55N306: C, 69.7; H, 8.7; N, 6.6. Found C, 69.4; H, 8.8; N, .6 go 6

I

6so 04 6 *6 6* 9 I5 Example Boc-Phe-PAla Amide of 2(S)-Amino-l-cyclohexyl- 3(R),4 (S)-dihydroxy-6--methylheptane Using the procedure of Example 33, but replacing Boc-PheAla with Boc-Phe-(3-pyrazoyl) alanine (Boc-Phe-PAla), gave the dflsired compound. Mass spectrum: 628.

Anal. calcd. for C 34 53N506 1/2H 2 0: C, 64.1; H, 8.5; N, 11.0. Found: C, 64.1; H, 8.3; N, 11.2.

25 Example 36 Ethoxycarbonyl-Phe-Leu Amide of 2(S)-Amino- 1-cyclohexl-3CR),4(S)-dihydroxy-6-methvlheptane 6 @6

S

S..

Us ing the procedure of Example 33, but replacing Boc-Phe-Ala with Boc-Phe-Leu, gave the desired compound. Mass spectrum: 576.

Anal. calcd. for C 32

H

53

N

3 0 6 C, 66.7; H, 19.3; N, 7.3. Found: C, 66.4; H, 9.5; N, 7.2.

Example 37 Boc-Phe-(SCH 3 )Cys Amide of 2( S)-Axnino-1-cyclohexyl- 3(R) ,4(S )-dihydroxy-6-meth'ylheptane Using the procedure of Example 33, but replacing Boc-Phe-Ala with Boc-Phe-(SCH 3 )Cys, gave the desired compound. Mass spectrum: 608.

Anal. calod. for C 32

H

53

N

3 0 6 S: C, 62.8; H, 8.8; N, 6.9. Found: C, 62.8; H, 8.9; N, 6.6.

Ts-(NExample 38 T-NMe,N 1 ,-Bn)-His Amide of 2(S)-Amino-icyclohexyl-3 ,4(S)-dihydroxy-6-methylheptane Using the procedure of Example 20, but replacing Boc-His with (N tosyl, N methyl, N imidazole benzyl)-His Ts-(N Me,N IM Bn)-His] (DuVigneau, V.; Behrens, O.K. J. Biol. Chem. 1937, 117, 27), gave the *6desired compound. Mass spectrum: 639.

Example 39 0Ds 20 Ethoxycarbon-yl-Phe-MeHis Amide Of 2(S)-Amino-icyclohexyl-3 ,4(S)-dihydroxy--6-methylhe-ptane To a stirred -7811C solution of the resultant .6..:compound of 'Example 38 (100 mg, 0.156 mrnol) in liquid 25 NH 3 (5 ML) ai d dry tetI.rahydrofuran (5 mL) was added sodium until a dark green/brown color persisted for :mint' tes. Solid, powdered NHi 4 C1 was then added, and the mixture was evaporated. The residue was suspended in water and extracted several times with chloroform.

The combined extracts were dried (Na 2 so 4 filtered, and evaporated to give the MeHis amide of 2(S)-aminol-cyclohexyl-3(R) ,4(S)-dihydroxy-6-methylheptane. The material was coupled to ethoxycarbonyl-Phe using to DCC/HQBT method described in Example 5 to give the desired product. Mass spectrum: =614.

V

I

Example 2( S)-t-Butyloxycarbonylamino-1-cyclohexyl- 7-methyloct-3-ene Using the procedure' of Example 13, but replacing isopentyltriphenylphosphoniwn bromide with isohexyltripheny'lphosphonium bromide, gave the desired product.

Example 41 2(S)-t-Butyloxyc rbonylamino-l.-cyclohexyl- ,(S)-dihydroxy--7-methyloctane Us ing, the procedure of Example 14, but replacing the res~ultant compound of Example 13 with the resultant compound of Example 40, gave the desired i compound.

Example 42 Boc-His Amide of 2(S)-Amino-l-cyclohexyl- 4(S)-dihyd'roxy-7-methyloctane Using the procedure of Example 20, bu~t replacing the diastereomer of Example 14 with the resultant compound of Example 41, gave the desired product. Mass spectrum: (M-iH) =495.

Anal. calcd. for C 26

H

6 N 0 5 1/2H 0: 26 464 C, 62.0; H, 9.4; 11.1. Found: C, 62.2; H, 9.4; N, 4 ~.0410.9.

Example 43 TBA-Phe-His Amnide of 2(S)-Amino-1-cyclohexcyl- 3(R) ,4(S)-dihydroxy-7-methyloctane Using the procedure of Example 15, but replacing the -resultant compound of Example 14 and Boc-Phe-His with the resul tant compound of Example 42 and t-butylacetyl(TBA)-Phe gave the desired compound.

Mass spectrum: (14-iH) =640.

Anal. calcd. for C 36

H

57

N

5 0 5 3/4H 2 0: C, 66.2; H, 9.0; N, 10.7. Found: C, 66.1; H, 9.1; N, 10.6.

Example 44 methylhex-3-ene Using the procedure of Example 13, but replacing isopentyltriphenylphosphonium bromide with isobutyltriphenylphosphonium bromide, gave the desired product. Mass spectrum: M+ 295.

Anal. calcd. for C 8

H

33

NO

2 1/4H 2 0: C, 72.0; H, 11.3; N, 4.7. Found: 71.7; H, 11.1; N, I

I

C C C C C 15 Example 2(S)-t-Butyloxycarbonylamino-l-cyclohexyl- 3(R),4(S)-dihydroxy-5-methylhexane Using the procedure of Example 14, but replacing the resultant compound of Example 13 with the resultant compound of Example 44, gave the desired compound.

Example 46 Boc-Phe-His Amide of 2(S)-Amino-l-cyclohexyl- 3(R),4(S)-dihydroxy-5-methylhexane Using the procedure of Example 15, but replacing the resultant product of Example 14 with the resultant product of Example 45, gave the desired product. Mass spectrum: 614.

Example 47 Ethoxycarbonyl-Phe-Leu Amide of 2(S)-Amino-lcyclohexyl-3(R),4(S)-dihydroxyhexane Following the procedures used to make the resultant compound of Example 36, but replacing isopentyltriphenylphosphonium bromide with propyl- 32 4 triphenyiphosphoniumn bromide, gave the desired product.

Mass spectrum: M+ 547.

Anal. calcd. for C 30

H

49

N

3 0 6 1/4H 2 0: C, 65.2; H, 9.0; N, 7.6. Found: C, 65.0; H, 8.9; N, 7.3.

Example 48 Ethoxycai~bonyl-Phe-Leu 'Am-,ide of 2(S)-Amino-icyclohexyl-3 Following the procedures used to make the resultant compound -of Exampole 36, but replacing isopentyltriphenylphosphonium bromide with phenethyltriph6nylphosphonium, bromide, gave the desired product.

0

S

*5 .4 i

OS

0*

S

S.

S

SO.

S

*OSSS.

4 0e 4* *6 S S S 0* Example 49" Boc-Phe-His Amride of 2(S)-Amino-l-cyclohexyVl- 3(R) ,4(S)-dihYdroxypentane Following the procedures used to make the resultant compound of Example 15, but replacing isopentyltriphenylphosphoniun bromide with ethyltriphenylphosphonium bromide, gave the. desired product.

Mass spectrum: 600.

Anal. calcd. for C' H Nl /4H 0: ?2 49

N

5 0 6 2 C, 63.6; H, 8.3; N, 11.6. Found: C, 63.6; H, 8.3; N, 211.5.

Example )-t-Butyloxylcarbonylamino-l-cyclohexyl- 3(S) 0 so 0 To a stirred -78 0 C solution of Boc-cyclohexylalanine methyl 'ester (35.0 g, 123 mmol) in anhydrous toluene (200 mL) was added diisobutylaluminumm hydride (140 1.5 M solution in toluene, 117 mL) at a rate to keep the internal temperatiare below -60 0 C. After stirring for an additional 20 minutes at -78 0 C, allyl magnesium chloride (184 mL of a 20 M solution in THF) was added. The mixture was allowed to stand at 0 0 C for 16 hours and was then q~uenched with methanol.' The mixture was diluted with ether and then washed seiquentially with citric' acid q) and brine. Drying (14gSO 4 and evaporating provided an oil which, was purified by silica gel chromatography to give the desired compound in 40% yield.

Example 51 2(S)-t-Bvtloycrbonylamino-l-cyclohexyl- 3(),4(S)-dihydroxyhex-5-ene An allylic oxidation using stoichiometric Se 2 and t-butyl hydroperoxide (Umbriet, M.A. and Sharpless, K.B. J. Am. Chemn. Soc. 1977, 99, 5526) was performed on the resultant product of Example 50 to give the desired product after silica gel chromatography, 0*~**gExample 52 Ethoxycarbonyl-Phe-Leu Amide of 2(S)-Amino--lcyclohexyl-3 Following the procedure of Example-1,5, but replacing the resultant product of Example '14 and Boc-Phe-His with the resultant product of Example 51 and ~*:':ethoxycarboriyl-Phe-Leu, gave the desired product, Anal.

calcd. for C 0

H

47 N 0 6 C, 66.03; H, 8.68; N, 25 7.70. Fouind: C, 66.10; H, 8.83; N, 7.43.

Examole 53 (N-Butyl, 4-OCH 3 )-Phenylalanine To a stirred DOC suspension of (4-OCF,, )-phenylalanine (1.00 g, 5.12 mmcl) and butyraldehyde (0.406 g, 110 M4%) in methanol (10 niL) was added sodium -cyanoborohydride (241 mg, 75M4%). The mixture was warmed to room temperature for 23 h and filtered.* The solid was washed with me 'hanol and suction dried to give 1. 07 g of the des.red product. Mass spect-lum: M' 251. *Anal.

0 34 Calcd f or Ci 14 2 1 N0 3 1/3" 2 0: C, 65.3; H, 8. 5; N, 5.4 Found- C, 65.1; 8.3; N, 5.6.

Example (-But 1, 4-OCH )Phe-His Amidb of 2(S)-Am"'no-iylohexyl-3(R),4(.S)- dihYdtoxy-6-methylheptane.

U .i n g the, procedure of Example 21, but replacing TBA-CHA with the resultant product of Example 53 gave the desired compound. Mass spectrum: 614.

Anal.- Calcd for C 3 4

H

55 N 5 0 5 .l/2 H 2 0: C, 65.6; 1; N, ft. 2. Fouhd,,V 65.3; H, 9.0, N, 11.3.

Example H-(4-0C0 3 aPr'-Leul'' Am ,ide of 2(S)-Amino-icycf2-hx" ,4(S)-dihydroxv-6-methylheptane.

Using the pr~cedure of Example 33, but replacing Boc-Phe-Ala with Cbz-(3-I,4- 0CM )"he-Leu provided the 6**protected iodinated prodi'ct. Deprotection and deiCodination was achieved by hydrog!enating 0.59 g in methanol (150 mL) with NaOAc. 3H 2 0 '(0.40 Rh/BaS0 4 (1.5 20% Pd/C (0.29 q) at 4 atmospheres H2 for 2.5 h. Filtration and evaporation provided a a:.a resi~iue which was partitioned between ethyl P.cetate and a.:'sat aHC, 3 The organic layer was washed with dilute 9a 2

S

2 0 3 and brine, dried, filtered, and 25 evaporated to give a solid. Recrystallization from

C

2

C

2 /hexan~e provided 260 .mg of the desired compound. HRMS: M Calcd 3b r C 30

H

52 N 3 0 534.3907. Measured: 534.3925.

Examnple 56 (N,N-Dimethyl ,4-methoxy)-Phe-Leu Amide of 2(S)-Amino-icyclohexyl-3(R) 4(S)-dihydroxy-6-methylheotane.

The resultant product of Example 55 (130 mg, 0.243 mmol) was hydrogenated (1 atmosphere H 2 with 10% Pd/C (39 mg) in methanol /formal in (12 mL/5 mL) for 8 h.

Filter/,!ng and evaporating (high vacuum) provided a 0esidt& e which was chromatographed on silica gel to -give 43 m~g of the desired compound., HRMS: _jcalculated for C 32

H

56

N

3 0 5 562.4220.

Measured: 562:4230.

'Zxample 57 H-Phe-Leu Amide of 2(S)-Amino-1-cYlhexl- S),4 -dihycdo6cy-6-methylheptane.

-17,o 1lowing the procedure of" Example 55, but repla %ng Cbz-(3-I,4-OCH b)Phe-Leu with Cbz-Phe-Leu and omitting NaOAc.3H 2 0 and 2.5% Rh/BaSO 4in the reduction step, provided the desired compound. Mass. spectrum: =504. Anal. Calod for CHN0 C, 69.1; H, VC 29

H

49

N

3 0 4 9. 8; N, Found: C, 90;H, 10.1; N, 8..

ExaMple 58 EN-(2-Cyanoethyl) Phe-Leu Amide of 2(S)-A-mino-icycJlohexyl-3 ,4,(S)-dihydroxy-6-methvlheptane, A suspension of the resultant compound of Example 57 (297 mg, 0,.590 mmol) in acrylonitrile (2 rnL) was -ref luxed _f r 3 days. Evaporation provided a. resi d ue 64which was dissolved in ethyl acetate, filtered, evaporated and chromatographed on silica I(dichloro- ~methane/methanol, 97.5/2.5) to give 162 mg (49 of the desired compound. Mass spectrum: 5517.

Anal. Calcd for C 2N 5 2 4 0 4 C, 69.0; 9.4; N, 10.1. Found: C, 68.6; H, 9.5; N, 9.9.

Example 59 (N-(3-Aminovrovvl) 3Phe-Leu Amide ot 2(S)-Amino-icyclohexyl-:3(R) ,4(,S)-dihydrox.y-6-meth-ylhe~tane.

The resultant compoutrA of,xample 58 (75 mg., 0.135 mmol) was hydrogenated (4 atmol~pheres H 2 over Raney Nickel (85 mg) in anhydrous metb-,,-'.nol/ammonia (20 ,ML/5 mLI for 3 h. Filtration and-- &aporation provided the desired product (68 mg). Mass spectrum: CMiH) +=561.

Example .(N,N-Dimethyl)Gly-Phe-His Amide of 2(S)-Arnino-l-cyclohexv 1-3 -dihydroxy-6--methylheptane.

Using the procedure of Example 56, but replacing the resultant product of Example 55 with the resultant produr-Ct of Example 64, gave the desired product. Mass 613.

Example 61 Cbz-B-Ala-Phe-His Amide of_2(S)-Axnino-l-cyclohexyl- ,4 (S )-dihydroxy-6-methylheptane.

Using the procedure of Example 21, but replacing THA-CHA with Cbz-B-Ala-Phe gave the desired compound.. Mass spectrum: 733. Analysis calculated for C H,-N 0 7 C, 65.5; H, 7.7; N, 11.5. Found- C, 65.2;, H, 7.7; N, 11.2.

Example 62 H-B-Ala-Phe-His Amide of 2(S)-Amino-l-cycloh 4(S)-dihydroxy-6-methylheptane Diacetic Acid Salt.

'levee"The resultant compound of Example 61 (1.00 g, 1.36 mmol) me in acetic acid (14 mL) was -hydrogenated at 1 atmosphere '4 with 10% Pd/C (0 50 g) for 3 h. Filtration, extraction 25 of the catalyst with acetic acid, and evaporation of the combined acetic acid solutions gave a residue which was dissolved in water (25 mL) and lyopholized to provide 891 mg of the desired product. Mass spectrum: 599 -(free base). Analysis Calculated for

C

36

H

58 N 6 0 9 '1/2H 2 0: C, 59.4; H, 8.1; N, 11.5.

Found: C, 59.3., H, 8.0; N, 11.2.

37 Example 63 Cbz-Sar-Phe-His Amide of 2(S)-Amino-l-cyclohexyl- 3(R),4(S)-dihydroxy-6-methylheptane, Using the procedure of Example 21, but replacing TBA-CHA with Cbz-Sar-Phe gave the desired compound. Mass spectrum: 733. Anal. Calcd for C, 64.8; H, 7.7; N, 11.3, Found: 65.0; H, 7.6; N, ll.S.

Example 64 H-S.ar-Phe-His Amide of 2(S)-Amino-l-cyclohexyl- 3(R),4(S)-dihydroxy-6-methylheptane Diacetic Acid Salt.

Using the procedure of Example 62, but replacing the resultant compound of Example 61 with the resultant compound of Example 63 gave the desired product. Mass spectrum: 599 (free base). Anal., calcd for C H NO C36 58H N60 .H 2 0: C, 58.7; H, 8.2; N, 11.4.

Found: 58.5; H, 8.1; N, 11.4.

4* C Example 20 Cbz-GABk-Phe-His Amide of 2(S)-Amino-l-cyclohexyl-3(R), 4(S)-dihydroxy-6-methylheptane.

Using the procedure of Example 21, but relacing TBA-CHA with Cbz-GABA-Phe (GABA is 4-aminobutyric acid) gave the S 25 desired compound.

Examole 66 H-GABA-Phe-His Amide of 2(S)-Amino-1-cyclohexyl-3(R),4(S) -dihydroxy-6-methyl-heptane Diacetic Acid Salt, Using the procedure of Example 62, but replacing the resultant compound of Example 61 with the resultant compound of Example 65 gave the desired product.

Example 67 Cbz-Isonipectoyl-Phe-His Amide of 2(S)-Amino-l-cyclohexyl ,4(S)-dihydroxy-6-methvlheptane.

Using the procedure of Example 21, but replacing TBA-CHA with Cbz-Isonipectoyl-E-,e gave the desired compound.

Mass spectrum: =773. Analysis calculated for C43 H60 N6 0 VH 2 0: C, 65.3; H, 7.9; N, 10.6.

Found: 65.4; H, 7.6; H, 10.5.

Examole 68 H-Isonipectoyl-Phe-His Amnide of 2(S)-Anmino-l-cyclohexyl-, 3(R) ,4(S)-dihYdroxy-6- methylheptane Diacetic Acid Salt.

Using" the procedure jof Example 62, but replacing the resultant compound of Example 61 with the resultant 1 compund of Example 67 gave the desired product. Mass spectrum: ="639 (free base).

S. **Example 69 *Cbz-D-Ala-P'fte-His Amide of 2(S)-Amino-l-cyclohexyl-3 CR), 4(S) -dihydroxy-6-methylheptane.

20 Using the procedure- of Example 21, but replacing TBA-CHA 0 o with Cbz-D-Ala-Phe gave the desired compound. Mass spectrum: =733. Analysis calculated for C 40

H

56 N 6 0 7 l5H 2 0: C, 63.2; H, 7.8; N, 11.0.

Found: C, 63.0; H, N, 11.0.

Example H-D-Ala-Phe-His Amide of 2(S)-Amino-l-cYclohexYl-3(R), :4(S)-dihydroxy-6-methylheptane Diacetic Acid Salt.

Using the procedure of Example 62, but replacing the resultant compound of Example 61 with the resultant too compound of Example 69 gave the desired product. Mass spectrum: 599 (free base).

39 Example 71 3-Benzyloxycarbonylamino-3-methylbutanoic Acid.

A solution of 2,2-dimethyl-3-carbomethyoxypropionic acid [LeMaul, Bull. Soc. Chim. Fr., 828 (1965), 20 g, 0.125 mol), diphenylphosphorylazide (34.3 g, 0.125 mol) and triethylamine was heated in toluene (150 mL) at 100 0

C

for 2 h. After cooling to 5°C, the toluene solution was washed successively with 0.5M HC1, aqueous NaHCO 3 and brine. Evaporation of the dried solution gave a residue which was chromatographed on silica gel eluting with 60/40 hexane- ether. There was obtained 13 g of methyl 3-isocyanato-3-methylbutanoate as a mobile liquid. A solution of this material in toluene (20 mL) was treated witi benzyl alcohol (13 mL) and the resulting mixture heated at reflux for 40 h. Evaporation of the toluene left a residue which was dissolved in methanol (125 mL) ,and then treated with a solution of NaOH (6.6 g, 0.165 mol) in 22 mL of water. After 5 h, the reaction mixture was partially evaporated, washed with ether and 20 acidified with 6N HC1. Extraction with methylene chloride and evaporation gave 21 g of the desired got product. NMR ,(300 MHz, CDCi 3 1.42 6H), 2.78 2H), 5.08 2H).

Example 72 Cbz-[(B,3-di-Me)-B-Ala]-Phe-OCH 3 A 4.0 g sample of 3-benzyloxycarbonylamino-3-methyl-

C

butanoic acid was coupled to phenylalanine methyl ester hydrochloride (3.43 g) using the mixed anhydride procedure described in Example 2. Purification of the crude product by flash chromatography eluting with 65/35 ether-hexane gave an 86% yield of product. NMR (300 MHz, CDC1 3 1.32 3H), 1.34 3H), 2.46 1H), 2.63 1H), 2.98 (dd, 1H), 3.09 (dd, 1H), 3.70 (s, 3H), 4.86 (dd, 1H), 4.97 H) 5.2 1H), 5.3 (s, 1H), 6.13 1H).

Example 73 Cbz--f (B,1-di-Me)-3-Ala)-Phe-OH To a 0 0 C solution of Cbz-[(13,1-di-Me)-1-Ala)-Phe-OMe (1.5 g, 3.63 mmol) in dioxane (15 mL) was added a solution of lithium hydroxide (0.174 g, 4.15 mmol) in water (7.5 mL). After stirring for 1 h at 0-5 0 C, the reaction mixture was diluted with cold water and extracted 2X- with ether. The aqueous portion was acidified with 6N HCl and extracted with ether. The organic extract was washed with brine and evaporated to give an 87% yield of product as a viscous liquid.

Example 74 Cbz-[(B1-di-Me)-B-Ala]-Phe-His Amide of 2(S)-Amino-i- 1f 5 cYclohex-yl-3(R) dihydrox-y-6-methylheptane.

Using the procedure of Example 21, but replacing TBA-CHA *.:with Cbz-E (3;S-di-Me)-I3-Ala3-Phe gave the desired compound. Mass spectrum: 71l. Anal. Calcd for C 42

H

60 N 6 0 7 .1/4H 2 0: C, 65.5; H, 8.0; N, 20 10.9. Found: C, 65.6; H, 7.9; N, 11.0.

Example H-f (3,B-di-Me)-1-Ala]-PhO-His Amide of 2(S) -Amino-icycJlohexyl-3(R) ,4(S)-dihydroxcy-6-methylheptane Diacetic Acid Salt.

Using the procedure of Example 62, but replacing the resultant compound of Example 61 with the resultant compound of Example 74 gave the desired product. Mass spectrum: 627 (free base). Anal. Calcd for C 38

H

62 N 6 0 9 'H 2 0: C, 59.7; H, 8.4; N, 11.0.

Found: C, 59.5; 8.4; N, 11.3.

Example 76 Cbz-Pro-Phe-His Amide of 2(S)-Arnino-l-cYclohexyl-3(R), 4(S)-dihYdroxy-6-methyl- heptane.

Using the procedure of Example 21, but replacing TBA-CHA with Cbz-Pro-Phe gave the desired compound. Mass spectrum: =759. Analysis calculated for C 42

H

58 N 6 0 7 .1/2H 2 0: C, 65.7, H, 7.7; N, 10.9.

Found: 65.7, H, 7.7; N, 10.9.

Example 77 H-Pro-Phe-His Amide of 2(S)-Amino-1-cyclohexcyl-3(R), 4(S)-dihydroxy-6--methylheptane Acetic Acid Salt.

Using the procedure of Example 62, but replacing the resultant compound of Example 61 with the resultant cqmpound of Example 76 gave the diacetic acid salt as a tacky solid. A portion of the di-salt was partioned *betwen satd. NaHCO 3 and dichJloromethane. The aqueous layer was further extracted with dichloromethane and the *combined organic layers were dried, filtered and evaporated to give the desired product. .Mass spectrum: 625 (free base). Analysis calculated for C C 36

H

56 N 6 0 7 2H 2 0: C, 60 H, 8.4; N, 11.6.

Found: C, 59.9; H, 7.9; N, 11.5.

Examvle 78 3-Benzyloxycarbonylamino-2 ,2-dimethylpropionic Acid.

3-Carbometho.-y-3-methylbutanoic acid ['Bull. Soc. Chim.

Fr., 828 (1965), 7.85 g, 0.049 moll was reacted with diphenylphospDhorylazide and triethylamine as described in Example 71. After heating the toluene solution for 5 h, benzyl alcohol (8 g) was added directly to the reaction mixture and heating- at ref lux was continued for 20 h. Work-up and purification as in Example 71 gave methyl 3-benzyloxycarbonylamino-2 ,2-dimethylpropionate.

NMR~ (300 MHz, CDC 3 1.2 6H), 3.3 2H), 3.68 3H), 5.1 2H), 5.22 1H). A sample of the methyl ester (6.21 g, 0.023 mol) was saponified with 3.1 g (0.78 mol) of NaOH in 100 mL ethanol/10 mL H 2 0 at room temperature for 48 h. Work-up as in Example 71 gave the desired product as a liquid. NMR (300 MHz, CDC1 3 1.23 6H), 3.32 2H), 5.10 2H), 5.27 1H).

Example 79 Cbz-[( a,a -di-Me)-3-Ala]-Phe-OCH 3 To a solution of 3-benzyloxycarbonylamino-2,2-dimethylpropionic acid (1.5 g, 5.97 mmol) in methylene chloride (13 mL) was added oxalyl chloride (0.757 g, 5.97 mmol) and dimethylformamide (30 ul). After stirring for I h at room temperature, the reaction mixture was cooled to 4 0°C and treated successively with phenylalanine methyl ester hydrochloride (1.29 g, 3.97 mmol) and N-methylmorpholine (1.81 g, 17.9 mmol). Stirring for 1 h at S" 0-5°C was followed by distribution between CH2C1 2 20 and 0.5 N HC1. The organic phase was washed with aqueous NaHCO 3 and brine and dried over MgSO 4 Evaporation of the sollent gave a residue which was purified by chromatography. There was obtained a 69% yield of product as a liquid. NMR (300 MHz, CDC1 3 25 1.11 3H), 1.12 3H), 3.05 (dd, 1H), 3.18 (dd, 1H). 3.23 1H), 3.24 1H), 3.75 4.82 (dd, 1H), 5.08 2H), 5.37 (broad t, 1H), 6.04 1H).

U.

Example Cbz-[( a, a -di-Me)-8-Ala]-Phe-OH.

The hydrolysis of the methyl ester was carried out by the procedure described in Example 71 to give the desired product in 90% yield as a viscous liquid.

Example 81 Cbz-t a, ch-di-Me)-51-Ala-Phe-His _AmiL'de of 2(S)-Aminocyclohexyl-3( (S)-dihydroxy-6-rnethylheptane.

Using the procedure of Example 21, but replacing TBA-CHA' with' Cbz-[ a ,a-di--Me)-13-Ala)--Phe gave the desired compound. Mass spectrum: =761.

Example 82 a-Di-Me)-B-Ala]-phe-His Amide of 2(S)-Amino-icyclohexyl-3(R) ,4(S)-dihydroxy-6-methylheptane Bits acetic acid salt.

Using-the compound from Example 81 and the procedure of Example 62 gave the desired product in 71% yield. Mass.spectrum: 627, Example 83 Cbz-Phe-His Amide'of 2(S)-Amino-l-cYclohexyl-3(R), 4(S) -dihydroxy-6-methylheotane :Using the procedure of Example 21 but replacing TBA-CHA 20 with Cbz-Phe gave the desired compound. Mass spectrum: 661.

Example 84 Phe-His Amide of 2(S)-Amino-l-cyclohexyl-3(R), 4 (S )-dihydroxy-6-methylheotane.

25 A solution of the product from Example 83 (180 mg, 0.273 mmol) in methanol .(50 mL) was hydrogenolyzed in a Parr Apparatus with 90 mg of 20% Pd/C and 4 atmospheres of *hydrogen. After the hydrogen uptake ceased, the catalyst was filtered and the filtrate evaporated to the desired product (90 mg, Mass spectrum: se i 527.

44 Example ai -Aminoisobutyryl-Phe-His Amide of 2(S)-Amino-1cyclohexyl-3(R),4(S)-dihydroxy-6-methylheptane.

A mixture of u-aminoisobutyric acid N-carboxy anhydride (10.9 mg, 0.085 mmol) and the product from Example 84 (44.6 mg, 0.085 mmol) in dimethylformamide (3 mL) was stirred at room temperature for 16 h. The dimethylformamide was evaporated in acuo and the residue was distributed between chloroform and water. The organic phase was dried and evaporated to a residue which was chromatogqaphed on silica gel eluting with methanol-chloroform mixtures. There was obtained 35 mg of the desired product. Mass spectrum: (M+H) 612.

Example 86 (Pyridin-3-yl-sulfonyl)-Phe-His "Amide of 2(S)-Amino-1cyclohexyl-3(R),4(S)-dihydroxy-6-methylheptane.

Using the procedure of Example 21, but replacing TBA-CHA 20 with (pyridin-3-yl-Tulfonyl)-Phe gave the desired product.

Example 87 (Pyrazin-2-yl-carbonyl)-Phe-His Amide of 2(S)-Amino-lcyclohexyl-3(R),4(S)-dihydroxy-6-methylheptane.

*goes: -4 0Using the procedure of Example 21, but replacing TBA-CHA with (pyrazin-2-yl-carbonyl)-Phe gave the desired product. Mass spectrum: 634. Anal. Calcd for C 34

H

47

N

7 0. 2/4H 2 0: C,64.0; H, 7.5; N, 15.4. Found: C, 63.9; H, 7.6; N, 15.2.

Examole 88 (Imidazol-4-yl-acetyl)-Phe-Leu Amide of 2(S)-Amino-lcyclohexyl-3(R),4(S)-dihydroxy-6-methylheptane.

Using the coupling conditions of Example 21 with 4-imidazoleacetic acid and the resultant product of Example 57 provided the desired product. Mass spectrum: 612. Analysis calculated for

C

34

H

53

N

5 0 5 1/2H 2 0: C, 65.9; H, 8.9; N, 11.3, Found: C, 65.9; H, 8.9; N, 11.3 Example 89 (Pyrrol-2-yl-carbonyl)-Phe-His Amide of 2(S)-Amino-1cyclohexyl-3(R),4(S)-dihydroxy-6-methylheptane.

Using the procedure of Example 21, but replacing TBA-CHA with (pyrrol-2-yl-carbonyl)-Phe gave the desired product. Mass spectrum: (M+H)r 621.

Example Allyloxycarbonyl-Phe-Leu Amide of 2(S)-Amino-1-cyclohexyl -3R),4(S)-dihydroxy-6-methyhe taine.

Using the procedure of Example 33, but replacing Boc-Phe-Ala with allyloxy carbonyl-Phe-Leu provided the desired product. Mass spectrum: 588. Anal.

20 Calcd for C 33

H

53 3 0 6 C, 67.4; H, 9.1; N, 7.2.

Found:--C, 67.6; H, 9.0; N, 7.1.

Example 91 3-Hydroxypropyloxycarbonyl-Phe-Leu Amide of 2(S)-Amino-lcyclohexyl-3(R),4(S)-dihydroxy-6-methylheptane.

4To a stirred 0C solution of the resultant compound of Example 90 (1.25 g, 2.13 mmol) in dry tetrahydrofuran (THF, 50 mL) was added 9-borabicycloE3.3.11- nonane S(9-BBN, 25.5 mL of a 0.5M solution in THP). The mixture was warmed to room temperature for 12 h and then cooled to 0 0 C. Water (15 mL) and 3M NaOH (4.5 mL) were added followed 2 min later by 30% H 2 0 2 (5 mL). The mixture was partitioned between brine (20 mL) and ethyl acetate (100 mL). The organic phase was washed (brine), dried (Na 2

SO

4 filtered, and evaporated to a thick oil. Recrystallization twice (dichloromethane/ether) provided 670 mg of the desired compound. Mass spectrum: 605. Analysis calculated for

CC

33

H

55

N

3 0 7 C, 65.4; H, 9.2; N, 6.9. Found: C, 65.4; H, 9.1; N, 6.8.

Example 92 Cbz-Gly Ester of the Resultant Compound of Example 91 (at 3-Hydroxypropyloxy Group), To a stirred 0 C suspension of the resultant compound of Example 91 (60 mg, 0.099 mol), Cbz-Gly-OH (20.7 mg, 0.099 mmol), and 4-dimethylaminopyridine (60 mg, 0.495 mmol) in dichloromethane (10 mL) was added ethyldimethylaminopropyl carbodiimide hydrochloride (38 mg, 0.198 mmol). The mixture was warmed at room temperature for 15 h and then diluted with dichloromethane and washed sequentially with 1M H 3

PO

4 satd NaHC033 33 4 3 and brine. Drying (Na 2

SO

4 filtering, and evaporating provided 57 mg of the desired compound. Mass spectrum: 797.

20 Example 93' H-Gly-Ester of the Resultant Compound of Example 91 (at 3-Hydroxypropyloxy Group).

The resultant compound of Example 92 (13 mg, 0.016 mmol) was hydrogenated (I -atmosphere H 2 with 10% Pd/C (4 4 25 mg) in methanol for 3 h. Filtration, evaporation and chromatography on silica (dichloromethane/methanol, 95/5- 90/10) provided 4 mg of the desired p roduct. HRMS: calcd for C 3 5

H

58

N

4 0 8 663.4333. Found: 663.4355.

Example 94 Lysine Ester of the Resultant CbmDound of Example 91 (at 3-Hydroxyropyloxy Group) Diacetic Acid Salt.

Following the procedure of Example 92 but replacing Cbz-Gly-OH with a,e-di- Cbz-Lys-OH provide the desired protected peptide. Hydrogenation according to the procedure of Example 93, but replacing methanol with acetic acid provide the desired compound.

Example Hemisuccinate Ester of the Resultant Compound of Example 91 (at 3-Hydroxypropyloxy Group).

Using the procedure of Example 92, but replacing Cbz-Gly with benzyl succinate provided the protected product.

Deprotection was achieved by following the procedure of Example 103 to give the desired product.

Example 96 Phosphate Ester of the Resultant Compoul Example 91 (at 3-Hydroxypropyloxy Grov 15 Using the procedure of Example 92, but\ Cbz-Gly with dibenzylphosphate provided the p d product.

Example 97 20 2(R,S),3-Dihydroxypropyloxycarbonyl-Phe-Leu Amide of 2(S) -Amino-l-cyclohexyl-3(R),4(S)-dihydroxy-6-methylheptane.

Following thepr6ocedure of Example 14, but replacing the resultant compound of Example 13 with the resultant 25 compound of Example 90, and heating the mixture at for 24 h, gave the desired product. Mass spectrum: (M+H) 622. Anal. Calcd for C 33

H

55 .1/2H 2 0: C, 62.8; H, 8.9; N, 6.7. Found: C, 63.0; H, 8.6; N, 6.7.

Example 98 Cbz-Gly Mono- and Diesters of the Resultant Compound of Example 97 (at the 3-Hydroxypropyloxy and 2, 3-Dihydroxypropyl Groups, Respectively).

Using the procedure of Example 92, but replacing the resultant compound of Example 91 with the resultant compound of Example 97, provided a mixture of the detsired mono- and die:3ters. separation was achieved by silica gel chromatography.

99 H-Gly-Ester of the Resultant Compound of Example 97 (at the 3-Hydro ropl-Group) Acetic Acid Salt.

Using the procedure of Example 93, but replacing the resultant compound of Example 92 with *the resul-tant monoester of Example 98' and replacing methanol with acetic acid, gave the desired product,

I

ssti

B

6S

G

0e.

9 6 4 4.

64...

4.6

S

I Example 100 H-Gly-Diester of the Resultant Compound of Example 97 (at the 2,3-Dihydroxypropyl.Grou-p) Diacetic Acid Salt.

Using the procedure of Example 93, but replaci~ the resultant compound of Example 92 with the resultant diester of Example 98 and replacing methanol with acetic acid, prov~ded! the desired coiarounzd.

Example 101 Ethoxycarbonvl.-(OBn)Thr-His Amide of 2(S)-Amino-icyc lohexyl-3 -dihydroxyv-6-methylhexane.

6660.0 *6 64. 6 Usipng the proaete of Exavi.IC 01L but replacing TBA-CHA 25 with ethoxycarbony-threonine benzyl ether E(OBn)Thr) gave the (iered compound. Mass spectrum, 616. Anal. Calcd for C 32

H

49

N

5 0 7 C, 62.4; H, N, 11.4. Found: 62.3; H, 8.0; N, 11.3.

Example 102 Benzyloxyacetyl-Phe-His Amide of 2(S)-Aznino-l-cyclohexyl- 3(R) ,4(S)-dihydroxy-6-methvlhe-otane.

Using the procedure of Example 21, but replacing TBA-CHA with benzyloxyacetyl-Phe gave the desired compound.

Mass spectrum: 6761. Analysis calculated for C 38

H

53

N

5 0 6 1/4H 2

C,

Found: 67.0; H, 7.9; N, 67,1; H, 7 9; N, 10.3.

I

Ex~ample, 1,03 Hydroxyacetyl-Phe-Hij. Aride of 2(.)-Amino-1-cyclohexyl- 3(R) ,4(S)-dihydroxy-6-methylheptane.

The resulltant compound of Examplt 1.02 (250 mg, 0.370 minol) in acetic acid (3.7 nIL) was hydrogenated at 1 atmosphere H 2 with 10% Pd/C (125 mg) for 23 h.

Filtration, extraction of -the catalyst with acetic acid, aid evaporation of the combined acetic acid solutions gave a residue whicli was partitioned between ethyl acetate and satd. aq. NaHCO 3 Exhaustive extraction of the acrueous phase with ethyl acetate, combinaticn of all organic layers, and eva.poration provided crude product wheach was recrystallized (ethyl acetate/methanol1 methylcyclohexane) to giire 157 mg of the desired a product. mass spectLuxn: 586. Anal. Calcd for C H 7 N 0 6 .H 0: C, 61..7; H, 8.2; N, 11.6.

Found: C, 62.1; 1H, 8.1; N, 11.4.

Examp 4 e 104 Acetyl-B--Ala-Phe-His Amide 2(S)-Aminio-l-cyclohexyl- 3(R) -dihydrox--6-metbylheptane.

Using the, procedure of Example 21, but replacing TBA-CHA 25 with Acetyl-B-Ala-',Phe provided the entire compound.

9) S q S.d 9 Exam-Dle 105 i-Bu-Pl-His Amide of 2(S)-Amino-l.-cyclohexyl-.3(R) d ihydr oxy- 6-methy lhvpt ane.

30 Using the procedure of Example 21, but replacing TBA--HA with O-isobutyl-L-3-phenyl lactic acid (i-Bu--Pl-OH) gave the desired compound.

Example 106 IsobutYrl-Ho-o--Phe methyl ester To a suspension of -amino-4-phenylbutyric acid (Homo-Phe) methyl ester hydrochloride (0.83 g, 3.61 mmol) in methylene chloride cooled in an ice bath was added successively isobutyric anhydride (0.57 g, 3.61 mol) and N-methylmorpholine (0.79 rnL, 7.22 mmol). After stirring for 30 min at 0-5 0 C, the reacb.ion mixture was distributed between methylene chloride and 0.5N HCI.

The organic layer was washed with aqueous NaHCO 3 and brine solution and then dried over MgSO 4 Evaporation of the solvent gave a solid residue which was triturated with hexane to provide 700 mg of product, mp 72-730.

Example 107 I sobutvrvl-Homo-Phe a 4* 0 0 as~ @0 Sao The hydrolysis of the methyl ester was carried out by the procedure described in Example 73 give the desired product in 90% yield.

Example i08 Isobutyryl-Homo-Phe '-His An~de off 2(S)-Amino-l-cyclohexyl- 3(R) ,4 S -dihydroxy-6-methylheptane, Using the procedure of Example 21, but replacing TBA-CHA with isobutyryl-homo- Phe gave the desired compound.

Mass spectrum: =612.

Example 109 2(S)-tM (4-Morpholinyl)carbonylloxy-3-phenylpropir-i.ic acid methyl ester.

To L-phenyllactic acid methyl 6ster (3.2 g) was added 150 mL of 12.5% phosgene in toluene and 25 drops of dimethylformamide. After stirring for 16 h at room temperature, the solvent-was evaporated and the residue chased several times with benzene, The resulting product was dissolved in methylene chloride (50 mL) cooled to 0 0 C and treated by dropwise addition with 3,86 g 044 mol) of morpholine. The reaction mixture was stirred for 2 h at 0-5 0 C and then distributed between 0. 5N H1-i and methylene chloride. The olganic phase was washed with aqueous NaHCO 3 and brine and evaporated to a residue. Flash chromatography on silica gel eluting with 2/1 ether-hexane gave a 65% yield of product. NI4R (300' MHz):' 3.08 (dd, 1H), 3.20 (dd, 1H), 3.8 5.19 (dd, 1H).

Example 110 0 (4-Morpholinyl)carbonylloxy-3-phenylpropionic acid.

:~15 Using the hydrolysis procedure of Example 73, the title compound was obtained in 90% yield.

~**Example 111 2(S)-C (4-Morpholinyl)carbonvlloxy-3-phenylp2ropionyvl-His Amide of 2(S)-Affino-l-cyclohexyl-3(R v4~)diyrx-6 -methylheTptane.

Using the procedure of Example 21, put replacing TBA-CH-A with the pDroduct from Example 110, gave the desired product in 60% yield. Mass spectrum: =642.

Example 112 2(S)-[C(4-Cbz-1-Piper~lzinyl)carbonylloxy]-3i: Phenyjlpropionic acid methyl ester.

Using the procedure of Example 109, but replacing morpholine with Cbz- piperazine,, gave the desired p7;oduct in 63% yield.

Example 113 2(S)-tfC4-Cbz-1-Pi-oerazinvl)carbonylloxyl)-3- P phenylpropionic acid.

Using the hydx,olysis procedure of Example 73 gave the desired product in 93% yield.

Example 114 2(S)-t[(4-Cb.z--Piperazinyl)carbonylloxy]-3phenylpropionyl-Phe-His Amide of 2(S)-Amino-1-cyclohexyl- 3(R) ,4(S)-dihydroxy-6-methylheptane.

Using the 'procedure of Example 21, but replacing TBA-CHA with the resultant compound from Example 113, gave the title compound. Mass spectrum: +=775.

Exampl 115 2(S)-E [(l-Piperazinyl)carbonylloxyl-3-phenylpropionyl- Seems:Phe-His Amide of 2(S)-Amino-l-cyclohexyl-3(R) p4(S)dihydroxy-6-methylheptane.

Using the procedure of Example 62 gave the title 15 compound in 85% yield. M.p. 158 0 -160 0

C.

Example 116 S(14-MAorpholinyl)carbonylj-Phe methyl ester.

A suspension of EI-phenylalanine methyl ester hydrochloride (6 g) in toluene (125 mL) was heated to 100 0 C while phosgene gas was bubbled into the reaction a. *mixture. After approximately 1-1/2-2 h, the mixture became homogeneous. The passage of phosgene was Scontinued for an additional 15 min, keeping the temperature at 90-100"C. The toluene was then evaporated and the residue chased several times with benzene. A f 1 ,g 03167 mol) sample>, of a-isocyanato-L-phenylalanine methyl ester was dissolved in 50 mL of methylere chloride and cooled to 0 0 C. Morpholine (2.76 InL, 0.03167 mol) dissolved in 5 mL of methylene chloride was, added dropwise. After 10 min at 0-5 0 C, the reaction mixture was* distributed between 0,5N HCl and methylene chloride. The organic layer was washed with aqueous NaHCO and dried over MgSO. Evaporation of the 3 4 solvent gave 7 g of product after trituration with 53 hexane, mp 90-910.

Example 117 C(4-Morpholinyl)carbonylI-Phe.

Using the procedure of Example 73 gave the title compound in 89% yield.

Example 118 [(4-Morpholinyl)carbonyl)-Phe-His Amide of 2(S)-Amino-lcyclohexyl-3(R),4(S)-dihvdroxy-6-methylheptane.

Using the procedure of Example 21, but replacing TBA-CBA with [(4-morpholinyl)carbonyl]-Phe, gave the desired compound. Mass spectrum: 641.

:Example 119 (Dimethylamino)carbonyll-Phe-His Amide of 2(S)-Amino-i- 15 cyclohexyl-3(R) ,4(S)-dihydroxy-6-methylheptane.

Using the procedures of Examples 116, 73, and 21, this compound was prepared. Mass spectrum: 599.

Example 120 [[Methyl-(2-hydroxyethyl)aminojcarbonyll-Phe-His Amide of 2(S)-iP-ino-1-cycloboxyl-3(R),4(S)-dihydroxy-6methylheptane.

Using the procedures of Examples 116, 73, and 21, the title compound was synthesized. Anal. calcd for

C

32

H

52

N

6 0 6 .1-1/2 H 20: C, 60.44; H, 8.45; N, 12.82. round: C, 60.36; H, 8.11; N, 12.77.

Example 121 [(l-Cbz-4-Piperazinyl)carbonyl]-Phe methyl ester.

Using the pr.ocedure of Example 116, but replacing morpholine with l-Cbz-piperazine, gave the desired product, mp 114-1150.

Example 122 (l-Cbz-4-Piperaz inyl )carbonyl I-Phe Using the procedure of Example 73 gave the desired product in 89% yield.

Example 123 (1-Cbz-4-Piperazinyl)carbonyl-Phe-His Amide of 2(S)- Amino-l-cyclohexyl-3(R) ,4(S)-dihydrox-y-6-meth_,heptane.

Using the procedure of Example 21, but replacing TBA-CXA with t (l-Cbz-4-piperazinyl)carbonyl)-Phe, gave the desired compound.

Example 124 t (l-Piperazinyl)carbonyll-Phe-His Amide of 2(S)-Amino-lcyclohexyl-3(R) ,4(S)-dihydroxy-6-methylheptane Bis- Acetic Acid Salt.

Using the procedure of Example 62 gave the desired compound in 90% yield. Mass spectrum: 640 (free base).

Example 125 E (4-Morpholinyl)carbonyll-(4-OCH )Phe methyl ester.

aUsing the procedure of Example 116 but replacing :H-Phe-OCH 3 HCl with L-tyrosine methyl ester methyl ether.HCl gave the title compound.

Example 126 (4-Morpholinyl)carbonyl ]-(4-OCH 3 )Phe-OH.

Using the procedure of Example 73 gave the title compound in 92% yield.

Example 127 £(4-MorPh2hlinl)carbonyl)-(4-OCH 3 )Phe-His Amide of 2(S) -Amino-l-cyclohexyl-3(R),4 (S)-dihdroxy-6methylheptane Using the procedure of Example 21, but replacing TBA-CHA with E(4-morpholinyl)carbonylj-(4-OCH )Phe gave the desired compound. Mass spectrum: 671.

Example 128 [4-(2-Oxopiperazinyl)carbonylj-Phe methyl ester.

Using the procedure of Example 116, but replacing morpholine with 2-oxopiperazine [Transition Met. Chem., *0*a11, 27 (1986)J gave the desired compound in 80% yield.

Example 129 (4-(2-Oxopiperazinyl)carbonyl3-Phe.

Using the procedure of Example 73 gave the desired compound.

Example 130 £4-(2-Oxopiperazinyl)carbonyl3-Phe-Hs Amide of 2(S)- Amino-l-cyclohexyl-3(R) 4(S)-dihydroxy-6-methylheptane.

Using the procedure of Example 21, but replacing TBA-CHA with oxopiperazinyl)carbonyll-Phe, gave the desired product in 60% yield.

S.r Example 131 E1-(4-Oxopiperidinyl)carbonyl3-Phe methyl ester.

Using the procedure of Example 116, but replacing morpholine with 4-pxopiperidine gave the desired compound.

56 Example 132 (l-(4-Oxopiperidinyl )carbonyl 3-Phe, Using the procedure- of Examp le 73 gave the desired compund in 91% yield.

Example 133 [l-(4-Oxo-piperidinyl)carbonyl--Phe-His Amnide of 2(S)- Amino-l-cyclohexyl-3(R).4 (S)-dihydroxy-6-methylheptane.

Using the procedure of Example 21, but replacing TBA-C-A with El-(4-oxopiperidinyl)ca, bonyl J-Phe, gave the desired product.

Boo&*: 41 0 Example 134 [1-(4-Hydroxypiperidinyl)carbonyl3-Phe methyl ester.

Using the procedure of Example 116, but replacing *415 morpholine with 4-hydroxypiperidine, gave the desired *44 compound.

Example 135 I l-(4-Hydroxypiperidinyl)carbonyl)-Phe.

Using the procedure of Example 73, gavd the desired .product in 82% yield.

Example 136 l-(4-Hydroxy-piperidinyl)carbonyl]-Phe-His Amnide of 2(S)- 25Amino-l-cyclohexyl-3 ,4(S)-dihydroxy-6-methylheptane.

Using the procedure of Example 21, but replacing TBA-CHA with I l-(4-hydroxypiperidinyl)carbonyl)-Phe, gave the desired compound in 56% yield.

Example 137 Il-(3-Hydroxypi~eridinyl)carbonyl]-Phe-His Arnide of 2(S)- Amino-l-cyclohexyl-3 ,4(S)-dihydroxy-6-met,,hylheptane.

Using the procedures described in Examples 116, 73 and 21, the title compound was synthesized.

Example 138 3-Carbomethoxy-3-phenoxypropionic acid.

A solution of 2-phenoxybutyrolactone [Dareman, Bull, Soc. Chim. Fr., 294 (1971), 4.96 g, 0,028 moll was added to methanol (125 mL) .containing 0.054 mol of sodium methoxide. After stirring for 3.5 hours at room temperature, the mixture was., quenched with 5 mL of acetic acid, and then distributed between ether and brine solution. The organic layer was washed with brine and evaporated to a residue (methyl-4-hydroxy-2-phenoxy- ,butyrate). A solution of this material in acetone (300 mL) was t;reated with Jones solution until the orange color persisted. The acetone was partially evaporated 15 and the residue was distributed between ether and brine solution. Evaporation of the dried ether layer gave the desired product as a waxy solid. NMR (300 NMR, CDC13): 3.02 2H), 3.78 3H), 5.11 1H).

Example 139 as** 20 SQ 3-[(4-Morpholinyl)carbonyl]-2-phenoxyprcionic acid methyl ester.

Using the mixed anhydride procedure described in Example 2, morpholine was coupled to 3-carbomethoxy-3-phenoxypropionic acid to give the desired product in 86% yield, mp 83-84 0 C. Anal. Calcd for C 15

H

19 NO5 C, 61,42; H, 6.53; N, 4.78. Found: C, 61.47; H, 6.50; N, 4.61.

Example 140 3- (4-Morpholinyl)carbonyl]-2-phenoxvprooionic acid.

Using the procedure of Example 73 gave the desired product in 59% yield, mp 150-15.1 0

C.

Example 141 3-E (4-Morpholinyl)carbonyl)-2(R,S)-phenoxypropionyl-His Amide of 2(S)-Amnino-i- cyclohexyl-3(R) ,4(S)-dihydrox-y-6methylheptane.

Using the procedure of Example 21, but replacing TBA-CHA with the resultant product of Example 140, gave the desired product as a mixture of R and S diastereomers.

Chromatography on silica (dichloromethane/methanol, 95/5) provided the less polar diastereomer (isomer A) and the more polar diastereomer (isomer Isomer A: mass spectrum: 642. Analysis calculated for C 34

H

51 N 5 0 7 .1/2H 2 0" C, 62.7; H, 8.0; N, 10.7.

Found: C, 62.7; H, 8.1; N, 10.3. Isomer B: Mass spectrum: 642. "Analysis calculated for

C

34

H

51 N 5 0 7 H 2 0: C, 61.9; 8.1; 10.6.

Found: C, 62.2; H, 7.8; N, 10.4.

Example 142

I

U

0 0 0 2(R,S)-(4-MorpholinylcarbonylmethyVl)- 3-Dhenylpropionic Acid.

4. a Ethyl a-barboxymethylcinnamate was prepared as reported (Cohen, S.G. and Milovanovic, A. Biochemistry, 1968, 3495) and hydrogenated according to the procedure of Exa'Ample 93. The resulting dihydrocinnamate was coupled to morpholine using the procedure of Example 21. Ester hydrolysis according to the procedure of Example 73 pDrovided- the desired compound. Mass spectrum: 278. Anal. Calcd for C 15H 13N 18H 20: C, 64.4; H, 6.9; N, 5.0. Found: C,64.4; H, 6.8; N, 4.9.

Exampie 143 2(R,S)-(4-Morpholinylcarbonylmethyl)-3-phenylpro-oionyl- His Amide of 2(S)-Amino-1-cyclohexyl-3(R) ,4(S)-dihydroxy- 6-methylheptane.

Using the procedure of Example 21, but replacing TBA-CHA 2(R,S)-(4-morpholinylcarbonylmethyl)-3-phenylpropionic acid provided t he desired product as a mixture of R and S diastereomers. Chromatog~raphy on silicaI (dichioro- methane/methanol, 95/5 90/10) provided the -iess polar diastereomer (isomer A) and the more polar diastereomer (isomer Isomer A: Mass spectrum: 640.' Anal. Calcd for C 35

H

53

N

5 0 6 .1/2H 2 0: C, 64.8; H, N, 10,8. Found: C, 65.1; H, 8.4, N, 10. 3. Isomer B: Mass spectrum: 640. Anal.Calcd for C 35

H

53 N 5 0.1/2H 2 0: C, 64'.8; H, 8.4; N, 10.8.- Found: C, 65.0; H, 8.3; N, 10.6.

ExamDle 144 N-(Benzyloxyacetyl~morpholine.

Using the mixed anhydride procedure described in Example 2, morpholine was coupled to benzyloxyacetic acid to 5555. give the desired product in 90% yield. ExamvDle 145 Methyl 2-benzyl-3-benzyloxy-3-( (4-moroholinyl)carbonylI pr pionate.

A -78 0 C solution of N-(benzyloxyacetyl)morph-)line (1 g,

SI

mmol) in THF (25 mL) was treated with potassium bis(trimethy3,silyl)amide (17 mL of a 0.5M solution).

After stirrlng for 10 min at -78 0 C, a solution of methyl 2-bromo-3- -phenylpropionate (8.5 mmol) in THF (5 mL) was 25 added dropwise. Stirring at -78 0 C for 30 min was followed by warming 'to 0 0 C. The reaction was then distributed between ether and brine solution. The organic layer was washed with brine and dried over MgSO4. Evaporation and flash chromatography on sil~ica gel gave the desired product in 63% yield.

Example 146 2-Benzyl-3-hydroxy-3-[ (4-morpholinyl)carbonyl 2 ProDionic acid.

Using the procedure of Example 84, the benzyl ether protecting group was removed by c-atalytic hydrogenolysis to give methyl 2-benzyl-3-hydroxy-3-[(4- morpholinyl) carbonylipropionate. Thet methyl ester function was hydrolyzed using the procedure in Example 73 to give the title compound.

Example 147 2-Benzyl-3-hydroxy-3-[ (4-morpholinyl )carbonyl )p-,ropionvl -His Amide of 2(S)-Amino-l-cyclohexyl-3(R),4(S)- 4jdihdroxy-6-methylheptane, Using the procedure of Example 21, but *eeplacing TBA-CHA with 2-benz yl-3-hydroxy-3-[(4-morpholinyl)carbonylI propionic acid, gave the d esired product in 51% yield.

Example 148 2-Hydroxy-3-[E(4-morpholinyllcarbonyll-oropionic acid acetonide.

A mixture of dl-malic acid (5 2,2-dimethoxypropa'-ie (100 mL) and catalytic p-TsOH was heated at 1000C for h. After cooling and evaj5oration the residual solid was recrystallized from carbon tetrachloride to give the 4 corresponding acetonide lactone. This material was 000:04 coupled to nmorpholine using the mixed anhydride procedure of Example 2 to give the title compound.

Example 149 Methyl 2-hydroxy-3-1 (4-mqr]?holinyl)carbony1 3propionate.

e 0 ago* A solution of 2-hydroxy-3-[4-(morpholi4nyl)-carbonylI *propionic acid, 1acetonide (5 g) iln. methyl alcohol (75 mL) was treated with 1 mL of concentrated sulphuric acid and the mixture was stirred for 24 h at: room temperature.

Partial evaporation of the solvent gave a residue which was distributed between either and brine solution. The ether layer was dried over MgSO 4 and evaporated to give the desired product.

9 *6 S. S *9 9* S 9.

S

'S

S

S..

S

.1500..

*56S

SO

Example 150 Methyl 2-anilino-3-( (4-morpholinyl)carbonyljpropionate.

The tL-rifluoromethanesulfonate of methyl 2-hydroxy-3- [4-morpholinyl)carbonyl) propionate was prepared by the method of Shiosaki Org. Chem., 4-6, 3230 (1981)]. A solution of this compound (7 minol) in methylene chloride mL) was added dropwise within 5 minutes at room temperature ,to a stirred solution of aniline? (14 mmol) in methylene chloride (25 mL), and stirring continued for 30 min at room temperature. The reaction mixture was filterd, the solution was washed with water, dried over Na 2 so 4 concentrated .and the residue purified by chromatography. Yield of product Example 151 2-Anilino-3-t (4-morpholin yl)carbonyllpropionyl-His Amnide 2(S)-Amino-l-cyclohexyl-3(R) ,4(S)-dihydroxy-6methyiheptane.

Using the product from Example 150 and the methods of Examples 73 and 21 gave the title compound.

Example 152 Ethyl 5-Acetamido--2(R,S)-b.en~zyl-4--oxopentanoate.

Ethyl a-carboxymethylcinnanate was prepared as reported (Cohen, S.G. And Milovanovic, A. Biochemistry, 1968, 3495) and hydrogen.ted according to the procedure of Example 93. The resulting aLcid was converted to the desired acetamidomethyl ketone using the methodology of Pfal~tz et al., (Tetrahedron Lett. 1984, 25, 2977: acid to acid chloride to cyanoketone followed by Zn/acetic acid/acetic anhydride treatment).

g* 0 62 Example 153 5-Acetamido-2(R,S)-benzyl-4-oxopentanoy.-His Amide of 2(S)-Amnino-l-cyclohexyl-3(R) ,4(S)-dihydroxy-6methylbeptane.

The resultant product of Example 152 was hydrolyzed according to the procedure of Example 73 provided the corresponding acid which s,.a's coupled in place of TBA-CHAaccording to the procedu..eb of Example 21. The desired product was obtained as 'an mixture which was separated by chromatography.

Example 154 (4-Morpholinyl)carbonylj-2--thiophenoxypropionic ac d methyl ester.

Using the procedure of Ex?'mr)'Ve 139, but replacing 3-carbomethoxy-,3-phenoxypropionic acid with 3-carbomethoxy-3-thiophenoxypropioni4' dd gave the desired product.

Example 1 (4-Morpholinyl)carbonyl]-2-(R,S'hI ophenoxypropionyl- His Amide of 2(S)-Amino--cclohexV.,- 3)R) ,4(S)-dihydroxy- 6-mrethylheptane.

as Using the procedures of Examples 73 and 21, t~hia title compound '<sprepared in 49% overall yield.

Example 156 2 (S )-t-Butyloxycarbonylaxnino-l-c clohe~al-3-hydroxy-6- 9 Ira 6methylheptan-4-one.

To a solution of resultant copudof Example 13 (8.50, 27.5 mmcl) in dry -THF (150 mL) were added 0504 (2.8 mL of a 2.5% solution- in t-butanol:9 ancd- N-methylmorpholine N-oxide (9.28 g, 68.7 mmol) After 4d the mixture was partitioned between either (?00 rCIL) and brine (100 mL) The aqueous layer was back-extratcted with either. (2 x lOOmL), and the combined organic phase was washed with 63 Na so 3 .0.1 M H PO 4 and brine., Drying (MgSC 4 and evaporating provided a residue (10.81 g) which was cbromatcgraphed on silica gel to remove the four diastereomeric diols from 0.7)0 g of the desired product. Mass spectrum: (M H) 342.

Exaple 157 Boc-Phe-His Amide of 2(S)-t-Butyloxycarbonylamino-lcyclohexyl-3-hydroXY-6-methylheptan-4-one.

The resultant product of Example 156 (22.0 mg, 0.645 mmol) was treated with 4 M 'HCl/dioxane for 6 hours.

Evaporation ,and drying under high vacuum provided th e corresponding amine hydrochloride which was dissolved in 641*dry dimethylformanide (DMF, 1.0 mL), tr~ated with Boc- 15 Phe-His (260 mg), N-methylmorpholine (J.142 mL), and I-hydrctybenzotriazole hydrate (261 mg), o 'ooled to -23 0 and then treated with 1-ethyl-3-(dimethyl- *aminopropyl) carbodiimide Hydrochloride (124 mg).

Evaporation after 16 lh provided a thick oil which was partitioned bbetween ethylacetate (60 mL) and saturated NaHCO 30m) The. organic phasq was washed with brine, dried CMgSO 4 and evaporated to give a residue which was chromatographed on silica gel (dichioroa rtethane/methanol) -to give 161 mg of the desiredr (M I 2.5 product. Mass spectrum: CM+=626. Anal.

calcd. for C 34

H

51 N50 6 C, ~53; H, 8.3; N, 11.2. Found: C, 65.6; H ,08.3; N, 11.2.

ExamL.e 158 Boc-Phe-His Amide (at N-2) of l-Cyclohexyl-2(S),4- S)-diamino-3-hydroxy-6-mehylhe Rtane.

Treatment of the resultant compound of Example 157 with hydroxylamine followed by reduction of the oxime over platinamn oxide gave the desired product.

Example 159 Ethoxycarbonyl-Phe-Leu Amide of 1-Cyclohexyl-2(S), 3(R,S) The resultant compound of Example 36 was acetylated using acetic anhydride and the corresponding 3-hydroxy- 4-acetoxy compound was isolated by silica gel chromatography. Oxidation to the 3-one using Jones reagent, deacetylization using sodium methoxide in methanol, and. reductive amination as in Example 158 gave the desired product.

q S *5

S

0W0~D

I~

q~ -6.

St.., S. a.

~S S Exal \1l6 0 Ethoxycarbonyl-Phe-His Amic± i-of 2(S)-Amino-l-phenyl-3 4(CS )-dihydroxy-6-methylheptane.

Using the procedure of Example 13, but replacing Boccyclohexylalanine methyl ester with Boc-Phe-OCH 3 and then following the procedures of Examples 14 and 29 gave the desired product.

ExamDle 161 Cyclic Carbonate of 2(S)-t-Butyloxycarbonylamino- 1-cyclohexyl-3(R) ,4(S)-dihy~roxy-6-methylheptane.

The 3(R),4(S)dias;tereomer of Example 14 was heated with N,N'-carbonyldiimidazole in benzene to give the desired compound in 86% yield.

Example 162 D-Se-PheHisamide of 2(S)-Amino-l-cyclohexyl-3(R) dihydroxy-6-methylheptane.

ae

B

a Following the procedure of Example 15, but replacing the resultant product of Example 14 with the resultant product of Example~ 161 and replacing Boc-Phe-His with Cbz-D-Ser-Phe-His gave the -desired N,O-diprotected material. N-deprotection following the procedure of S Example 62 followed by O-dep ;otection with 0.5M NaOH in aq.dioxane, gave the desired compound.

Example 163 2(S)-Isobutyrylmercapto-3-phenylpropionyl-Phe-H is Amide of 2(S)-Amino-1-cyclohexyl-3(R),4(S)-dihydroxy-6rethylheptane.

S(+)-2-mercapto-3-phenylpropionic acid was prepared as described (Acton, N and Komoriya, A. Organic Preparation and Procedures Int. 1982, a4, 381-392.) and acylated with isobutyric anhydride. Replacing TBA-CHA with this acid and using the procedure of Example 21, gave the titled compound.

Example 164 e (2-Aminoethyl)mercato-3-phen-propionyl-Phe-His A- Amide of 2(S)-Amino--cyclohexyl-3(R),4(S)-dihydroxy-6- O methylheptane.

(2-Aminoethyl)mercapto)-3-phenylpropionic acid was made using literature methodology (Acton, N. and V Komoriya, A. Organic I~rejarations and Procedures Int.

1982, 14, 381-392.) Replacing TBA-CHA with this acid and using the procedure of Example 21, gave the titled compound.

Example 165 (2S ,3R, 5R)-2-(t-Buty loxycarbonylamino)-3-hydroxy- 7-methyl--phenyloctane-5-carboxylic Acid Lithium Salt A soluation of 27.1 mg (0.075 mmol) of (3R,5Rl'S)-5- -butyloxycarbonylamino)-2-phenylethyl)-3-isobutyldihydrofuran-2-(3H)-one Kempf, J. Org. Chem. 1986, 51, 3921) in 1 m.L of dioxane was treated with 185 ul (0.092 mmol) of LiOH (0.5 M in H 2 0) and- stirred at ambient temperature for, 8 h. Removal of the solvent in vacuo gave the desired compound as a white solid.

66 Example 166 (2S,3R,5R)-3-(t-Butyldimethylsilyloxy)- 2-(-t-butyloxycarbonylamino )-7-methyl-1-phenvloctane- Acid t-Butyldimethylsilyl Ester A solution of the resultant compound of Example 165 (0.075 mmcl), 42 mg (0.28 mmcl) of t-butyldimethylsilyl chloride and 31 mg (0.45 mmcl) of imidazole in 0.8 mL/ of dimethylformamide was allowed to stand at ambient temperature for 2 days. Removal of the solvent in vacuo gave the crude desired compound.

Example 167 (2S,3R,5R)-3-(t-Butyldimethylsilyloxy)- 2- (t-butyloxycarbonylamino )-7-methyl- %too: -pheniyloctane--5-carboxylic Acid Lithium Salt fi 15 A solution of the crude resultant compound of Example 166 (0.075 mmcl) in 2 mL of dioxane was treated with 0.6 mL (0.3 mmol)- of LiOH. (0.5 M in H 2 0) and allowed *to stir at ambient temperature for 2 days., After 0* 20* removal* of the solvent, purification by flash column chromatography using 3% methanol/chloroform gave 18.3 mg of the desired compound (R f 0.10, 2% methanol/chloroform).

Example 168 (2S,3R,5R,.S, 9R,10S)-7-Aza- -3-(t-butyldimethylsilyloxV)-2-(t-butvloxycarbonyl amino cyclohexylmethyl 12-Tnethyl-l-phernrltr idecane .5 Using the coupling procedure of Example .24but replacing Boc-Phe-His-OM with the resultant compound of Example 167 gave the desired compound in 62% yield af ter purification by MPLC using4 6:1 hexane/ethyl acetate (R f 0.50, 2:1 hexane/ethyl acetate).

67 Example 169 (2S,3R, 5R, 8S ,9R, 1OS)-7-Aza- 2-(t-butyloxycarbonylamino)-8-(cyclohexylmethyl)- 5-isobutyl-12-methyl-l-phenyl-3 ,9 0-tr2., hydroxytridecane A ,solution of 16.5 mg 023 mmol) of the resultant compound of Example 168 in 1' mL of tetrahydrofuran was treated with 70 mIL (0.07 mmol) of tetra-n-butylarmmnonium fluoride (1 M. in tetrahydrofuran) and allowed tp stir at ambient temperature for 16 h. After concentration in vaduo, separation by MPLC using 2:1 hexane/ethyl acetate gave 10.5 mg of the desired compound as a white crystalline solid. Mass spectrum: (M H) 605.

Example 170 15 Cbz-6--aminohexanol--( 4-methoxy)phenylalanine Benzyl Ester **.Using the procedure of Example 72 but replacing *.3-benzyloxycarbonylamino-3-methylbutanoic acid with 6-(Cbz-amino)-n-caproic acid and replacing phenylalanine methyl ester with (4-methoxy)phenylalanine benzyl ester gave, after purification by flash column chromatography using 9:1 chiloroform/ethyl acetate, a 38% yield of the desired compound.

Example 17~ :Cbz-6-aminohexanoyl-( 4-methoxy)ohenylalanine A solution of 2.66 q (5 inmol) of the resultant compound of Example 170 in 60 m.L of tetrahydrofuran was cooled to 0 0 C, treated with 0.63 g (15 mmol) of LiOH in 30 niL of H 0 1and allowed to stir for 2 h. After concentration of the solv 6nt, the 'mixture was partitioned between and ether, acidified, extracted with ethyl acetate, dried over MgSO 4 and concentrated to give 1.55 g of the desired compound.

Example 172 Cbz-6-aminohexano 4-methoxy)Phe-His Amide of S,3R,4S)-2-Amino-1-cyclohexyl- 3, 4-dihydroxy-6-methylheptane.

Using th( procedure of Example 21 but replacing TBA-CHA with the resultant compound of Example 171 gave, after recrystallization from ethyl acetate, a 79% yield of the desired compound. Mass spectrum: (M H) +=805.

Example 173 6-Aminohexanoyl-( 4-methox~y)Phe-His Amide of (2S,3R,4S)-2-Anino-l-cyclohexyl-3 ,4-dihydroxy- 6-methylheptane Diacetate Salt.

A mixture of 0.97 g (1.2 mmol) of the resultant compound of Example 172 and 0.20 g of 20% palladium on carbon in 150 mL of 95% aqueous acetic acid was shaken in a Parr *Apparatus under four atmospheres of H2 After 2.

filtration to remove catalyst, the solution was concen- 0 trated' in vacuo, diluted with 75 mL of H820, and concentrated by lyophilization to give 0.86 g of the desired compound as a white solid. Mass spectrum:

(M+H)

t 671.

Examp le 174 E(4-Mor-pholinyvl)carbonyl-D-Phe-His Amide of 2(S)-Amino-1-cyclohexyl-3(R) ,4(S)-.dihydroxy- 6-methylheptane.

*.Using the procedures of Examples 116, 117 and -118 but replacing L-Phe-OCH 3 HCl with D-Phe-OCH 3 HC1, gave the title compound. Mass spectrum: (M H) 30 641.

ExamvDle 175 Ethyl Hydrogen ct, c-dimethylbenzyl)malc/nate.

-dimethylbenzyl)malonate was prepared by the congugate addition of phenyl magnesium bromide to *o C C

C

C.

C

S

me..

C

C. C diethyl isopropylidenemalonate as described by C. Holmberg [Liebigs Ann. Chem., 748 (1981)], A solution of this diester (42.1 g, 0.15 mole) in ethanol (100 mL) was treated by dropwise addition with a solution of potassium hydroxide (8.48 g, 0.13 mole) in 100 mL of ethanol. After heating at 90 0 C for 1 h and at 0 C for 20 h, the reaction mixture was evaporated on the rotary evaporator to a residue. The residue was diluted with water and extracted with ether to remove unreacted starting material. The aqueous phase was coolk c to 5 0 C, acidified to pH 3, with 6N HC1 and extracted with methylene chloride. The organic layer was washed with brine solution and dried over magnesium sulfate. Evaporation of the solvent gave 27.3 g (84%) of liquid product. NMR (CDC13): 1.05 (3H, t), 1.6 (6H, 3.78 (1H, 3.96 (2H, 7.2-7.4 (5H, m).

-'j Example 176 Ethyl [(4-morpholinyl)carbonyllamio]- 3,3-dimethyl-3-phenylpropionate. To a solution of ethyl hydrogen a a -dimethyl enzyl) malonate (4 g, 0.016 mole) in toluene was added triethylamine (2.23 mL, 0.016 mole) and diphenylphosphoryl azide (4.4 g, 0.016 mole). The reaction 25 mixture was .heated at ld00C for 2.5 h, cooled to 5 0

C,

and treated with 1.4 mL (0.016 mole) of morpholine.

After stirring overnight at room temperature, the toluene solution was washed successively with 1N HC1 and aqueous sodium bicarbonate solution. The dried organic solution was evaporated to a residue which was purified by column chromatography on silica gel. There was obtained 3.7 g of product after trituration with hexane, mp 93-94"C.

Anal. calcd. for C 18

H

26

N

2 0 4 C, 64.65; N, 7.84; N, 8.38.

Found: C, 64.72; H, 7.95; N, 8.33.

I

C*

Example 177 ((4-Morpholinyvl)carbonyllamino)- 3, 3-dimethyl-3-phenylpropionic Acid.

0 A solutio,', of the product form Example 176 (2 g, 5.99 mmole) in dioxane (10 mL) was treated with 0.26 g mrnol) of sodium hydroxide in 5 mL of water. After stirring for 16 h at 35 0 C, the reaction was worked up as described in Example 175 to give a 93% yield of product.

Example 178 £(4-Morpholinyl)carbonyl]amino]- 3 ,3-dimethyl-3-phenylpropionyl-His Amide of 2(S)-Amino-l-cyclohexyl-3(R) ,4(S)-dihydroxy- 6-methylheptane.

4 a.

4 .4 44 4.

4.44 4 i 444 .4 4 4*s 4 444444

S

44 4~ 9.

44 4.

4. 4 4* K 4.

The product f rom Example 20 was deprotected with HCl! 15 methanol and coupled to the product from. Example 177 using the procedure described in Example 5 but modified as follows. HOBT was not used in the coupling and the reaction time Was 20 h. There was obtained an 80% yield ~of the desired product. Mass spectrum: (M +H) 669.

Example 179 H-Isonipecotvl-(4-OCH 3 )Phe-His Amide of 2(S )-Amino-l-cyclohexYl-3(R) ,4(S)-dihydroxV- 6-methylhep~tane Diacetic Acid Salt.

Using the procedure of Examples 67 and 68, but replacing Cbz-isonipecotyl-Phe 'with Cbz-isonipecotyl-( 4-OCR 3 Phe gave the desired product. ,Mass spectrum: (M H =669 (free base).

Example 180 H-[(B,B-di-Me)-V--Ala1-(4-OCR 3 )Phe-His Amide of 2(S)-Amino-l-cyclohexyl-3 4(S)-dihydroxy- 6-methylheptane Diacetic Acid Salt.

Using the procedures of Examples 74 and 75, but replacing Cbz-[(B,B-di-Me)-8-Ala)-Phe with Cbz-[(B,B-di-Me)-S-Ala]-(OCH3)Phe gave the desired product. (M 657 (free base).

Example 181 2(S)-t-Butyloxycarbonylaminc-l-cyclohexyl- 3(R)-hydroxy-6-methylheptan-4-one To a stirred -63 0 C solution of oxalyl chloride (784 mg, 6.18 mmol) in dry dichloromethane (15 mL) was.

added dry dimethylsIfpoxide (708 mg, 9.06 mmol) dropwise over 5 minutes After another 5 minutes, Boc-cyclohexylalaninol (1..06g, 4.12 mmol) in dichloromethane mL) was added dropwise over 5 minutes, and 5 minutes later, triethylamine (1.67 g, 16.48 mmol) was added similarly. ZnI 2 (300 mg, 0.94 mmol) was added over 15 minutes. After stirring for 2 minutes, trimethylsilyl cyanide (1.43g, 14.42 mmol) was added and the mixture was warmed to room temperature for I hour. The mixture i. was then cooled to 0°C and isobutylmagnesium chloride (22.0 mL of a 2 M soln. in ether) was added. After warming to room temperature for 4 hours, the mixture was poured into 1 M H 3

PO

4 (40 mL)/ice (50 mL) and extracted with ethyl acetate. The combined organic Sphase was washed sequentially with 1 M H 3

PO

4 water, 3 4 satd. NaHCO 3 and brine. Drying (MgSO 4 filtering, and evaporating provided 1.75 g of an oil which was dissolved in THF (75 mL) 'and treated with 1 M

H

3

PO

4 (25 mL) for 18 hours at 5°C. The solution was partitioned between ethyl acetate/brine, and the resulting- organic phase was washed sequentially with brine, satd. NaHCO 3 and brine. Drying 0 4 filtering, and evaporating provided the desired product (1.39 g, 99%) which was used directly in the next step.

-72 Example 182 2(S)-t-Butvloxvcarbonylami no-l-cyclohexyl- 3(R) .4(S)-dihydroxy-6-methvlheptane To a stirred solution of 2(S)-t-Butyloxycarbonylamino-l-cyclohexyl-I 3(R)-hydroxy-6-methylheptan-4-one (200 mg, 0.586 mmol) in THF (10 mL) was added NaBH 4 (22 mg, 0.586 mmol). After 2 hours, the solvent was evaporated and the residue was partitioned between ethyl acetate and brine. The organic phase was washed (brine), dried (MgSO 4 filtered and evaporated. The residue was recrystallized from methylcyclohexane to give 76 mg of the desired product. M.p. 130-131*C. The mother liquo 'r was chromatographed (silica gel, ether/hexane) to afford 43 mg more.

Example 183 (2S.3R.5R.8S.9R.10S)-7-Aza-2j4,(t-Butyloxycarbonvylamino)- -8-(cvclohexvlme-thvl '-methvl-5-(4-pentenyl *0a*I: -l-ph-Qny 3.'.10 ri hvdroxvtri decane ~*Using the procedures of ExamRIPie-165-169, but replacing 5-E(t-butyloxycarbonylamino)-2-phenylethyl )-3-isobutyldihydrofuran- 2-(3Hone with (3.R,5R,1 '_)-5-1(.t-butyloxycarbonylamino)-2-phenylethyl 2-3-(4- 06610 pentenyl)dihydrofuran-2-(31)-one Kempf, J. Org. Chem. 1986, 51, .*.3921) gave the desired compound in 52% yield after purification by MPLC using 2:1 he-x ane/ethyl acetate. Mass spectrum: 617.

The dLscribed compounds can be used in the form of salts derived from inorganic or organic acids. These salts include but are not limited to the following: acetate, adipate, alginate, citrate, aspartate, benzoate, benzene sulfonate, bisulfate, butyrate, camphorate, camphorsulfonate, digluconate, cyclopentanepropionate, dodecylsulfate, ethanesulfonate, TCW/KXW: 1052v 73 glucoheptanoate, glycerophosphate, hemisulfate, heptonate, hexanoate,.

fumarate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxy-ethanesulfonate, lactate, maleate, methanesulfonate, nicotinate, 2-naphthalenesulfonate, oxalate, pamoate, pectinate, persulfate, 3-phenylpropionate, picrate, pivalate, propionate, suc'inate, tartrate, thiocyanate, tosylate, and undecanoate. Also, the basic nitrogen-containing groups can be quarternized with such agents as loweralkyl halides, such as methyl, ethyl, propyl, arid bityl chloride, bromides, and iodides; dialkyl sulfates like dimethyl, diethyl, dibutyl., and diamyl sulfates, long chain halides such as decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides, aralkyl halides like benzyl and phenethyl bromides, and others. Water or oil-soluble or dispersible products are thereby obtained.

Examples of acids which may be employed to form pharmaceutically acceptable acid addition salts include such inorganic acids as hydrochloric acid, sulphuric acid and phosphoric acid and such organic acids as oxalic acid, maleic acid, succinic acid and citric acid. Other salts include salts with alkali metals or alkaline earth metals, such as sodium, potassium, calcium or magnesium or with organic bases.

The described compounds can also be used in the form of esters.

Examples of such esters include a hydroxyl-substituted compound of formula I which has been acylated with a blocked or unblocked amino acid residue, a phosphate function, or a hemisuccinate residue. The amino acid esters of particular interest are glycine and lysine; however, other amino acid residues can also be used. These esters serve as pro-drugs of the compounds of the present invention and serve to increase the solubility of these substances in the gastrointestinal tract. The preparation of the pro-drug esters is carried out reacting a hydroxyl-substituted compound of formula I with an activated amino acyl, phosphoryl or hemisuccinyl derivative. The resulting product is then deprotected to provide the desired pro-drug ester.

00. TCW/KXW:1052v 74 The described compounds possess an excellent degree of activity and specificity in treating renin-associated hypertension in a host. The ability of these compounds to inhibit human renal renin can be demonstrated in vitro by reacting a selected compound as varied concentrations with human renal renin, free from acid proteolytic activity, and with renin substrate (human angiotensinogen) at 37 0 C and pH 6.0. At the end of the incubation, the amount of angiotensin I formed is measured by radioimmunoassay and the molar concentration required to cause 50% inhibition, expressed as the IC 50 is calculated. Nhen tested in accordance with the foregoing procedure, these compounds demonstrated IC 50 's in the range pf 5 to 11 0 M as seen in Table I.

Se S. e so

S

S..

TCW/KXW:1052v Table I Example EFcample Num~ber ICpso (nM) Number IC 5 0 (nM4) 34000 63 0.45 6 50 64 3 1.5 67 0.8 16 70 68 1 17 35 69 0.81 18 95 70 21 2 74 0.7 22 1.5 75 0.4 23 10 76 ~j 24 2 77 0.98 20 81 0.6 26 1.5 82 0.6 27 7 83 0.6 28 80 84 *29 0.6 85 0.4 0.75 87 0.55 531 1 88 0.6 1532 2 89 1 33 5 90 0.4 34 1.5 91 0.3 1 92 36 0.4 93 0.55 37 0.5 97 0.3 39 2 101 43 5 102 0.6 *46 1.5 103 1 :47 1 108 0.55 49 2 111 54 0.95 114 1.3 2 1 15 1 56 5.5 118 577.5 124 0.65 58 7 127 0.75 61 0.55 141 62 2 143 0.3 169 6.0 178 2 173 0.9 179 1 174 12 180 0.8 183 12

U

76 These compounds may also be used with one or more antihypertensive agents selected from the group consisting of diuretics, and/or B-adrenergic blocking agents, central nervous system -acting agents, adrenergic npiron blocking agents, vasodilators, angiotensin I converting enzyme inhibitors, and other antihypertensive agents.

Total daily dose administered to a host in single or divided doses m7y be in amounts, for example, from 0.001 to 10 mg/kg body weight daily and more usually 0.01 to 1 mg. Dosage unit compositions may contain such amounts of submultiples thereof to make up the daily dosage.

The amount of active ingredient that may be combined with the carrier materials to produce a single dosage form will vary depending upon the host treated and the particular mode of administration.

It will be understood, however, that the specific dose level for any particular patient will depend upon a variety of factors including the activity of the specific compound employed, the age, body weight, general health, sex, diet, time of administration, route of administration, rate of excretion, drug combination, and the severity of the particular disease undergoing therapy.

These compounds may be administered orally, parenterally, by inh\la tion spray, rectally, or topically in dosage unit formulations containing conventional nontoxic pharmaceutically acceptable carriers, adjuvants, and vehicles as desired. The term parenteral as used herein includes subcutaneous injections, intravenous, intramuscular, intrasternal injectable, or infusion techniques.

"5 Injectable preparations, for example, sterile injectable aque Ior oleagenous suspensions may be formulated according to the known art using suitable Go *Q TCW/KXW:1052v 'Iwo 77 dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a nontoxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil may be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid find use in the preparation of injectables.

Suppositories for rectal administration of the S 15 drug can be prepared by mixing the drug with a suitable nonirritating excipient such as cocoa butter and I polyethylene glycols which are solid at ordinary temperatures but liquid at the rectal temperature and 1 will therefore melt in the rectum and release the drug.

20 Solid dosage forms for oral administration may include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound may be admixed with at least one inert diluent such as sucrose lactose or starch. Such dosage forms may 25 also comprise, as is normal practice, additional substances other than inert diluents, lubricating agints such as magnesium stearate. In the case of capsules, tablets, and pills, the dosage forms may also comprise buffering agents. Tablets and pills can 30 additionally be prepared with enteric coatings.

Liquid dosage forms for oral administration may include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs containing inert diluents commonly used in the art, such as water.

Such compositions may also comprise adjuvants, such as wetting agents, emulsifying and suspending agents, and 78 sweetening, flavoring, and perfuming agents.

The foregoing is merely illustrative of the invention and is not intended to limit the invention to the disclosed compounds. Variations and changes which are obvious to one skilled in the art are intended to be within the scope and nature of the invention which are defined in the appended claims.

aie

G'.

A

ip

Claims

1. A compound of the formula: OP 2 P 1 NH R' OP 3 wherein R' is loweralkyl, vinyl or arylalkyl as hereinbefore defined; S P2 and P 3 are independently either hydrogen or an 0-protecting group; and P 1 is hydrogen or an N-protecting group; or an acid addition salt S* B thereof.

2. The compound of Claim 1 wherein R' is -CH CH(CH 3 2

3. A compound of the formula: OP 2 N CH 3 NH OP 3 CH 3 oB t wherein P 1 is hydrogen or an N-protecting group and P 2 and P 3 are 10 independently either hydrogen or an 0-protecting group; or an acid addition salt thereof.

4. 2(S)-t-Butyloxycarbonylamino-l-cyclohexyl-3(R), 4(S)- dihydroxy-6-methylheptane. l 't 2(S)-Amino-l-cyclohexyl-3(R), 4(S)-dihydroxy-6-methylheptane, or acid addition salts thereof.

6. A compound as defined in Claim 1 and as herein described with reference to any one of Examples 14, 15, 45, 51 or 182 or as described with reference to Scheme I or Scheme II. DATED this SIXTEENTH day of MARCH 1993 Abbott Laboratories Patent Attorneys for the Applicant SPRUSON FERGUSON 1654a/ii