WO1988007053A1 - Renin inhibitors containing a cyclopropyl amino acid and/or a cycloalkyl transition-state analogue - Google Patents

Abstract

The present invention provides novel renin-inhibiting peptides having non-cleavable transition state inserts corresponding to the 10,11-position of a renin substrate (angiotensinogen) and having at least one cyclopropyl amino acid in place of an amino acid residue corresponding to position 6, 8, 9, 12, 13 or 14 of the renin substrate; and/or a cycloalkyl amino acid non-cleavable transition state insert in place of an amino acid residue corresponding to position 10 and 11 of the renin substrate. Such inhibitors are useful for the diagnosis and control of renin-dependent hypertension and other related deseases.

Description

Renin inhibitors containing a cyclopropyl amino acid and/or a cycloalkyl transition-state analogue.

DESCRIPTION BACKGROUND OF THE INVENTION

The present invention provides novel compounds. More particularly, the present invention provides novel renin-inhibiting peptide analogs. Most particularly, the present invention provides renininhibitory compounds having non-cleavable transition state inserts and having at least one cyclopropyl amino acid and/or a cycloalkyl non- cleavable transition state insert. The renin inhibitors provided herein are useful for the diagnosis and control of renin- dependent hypertension and other related diseases.

Renin is an endopeptidase which specifically cleaves a particular peptide bond of its substrate (angiotensinogen), of which the N-terminal sequence in equine substrate is for example: Renin

↓ Asp-Arg-Val-Tyr-lie-His-Pro-Phe-His -Leu-Leu-Val-Tyr-Ser- IA

1 2 3 4 5 6 7 8 9 10 11 12 13 14

as found by L.T. Skeggs et al, J. Exper. Med. 106, 439 (1957). Human renin subsrrate has a different sequence as recently discovered by

D.A. Tewkesbury et al , Biochem. Biophys. Res. Comm. 99, 1311 (1981).

It may be represented as follows:

Renin ↓

-Val-Ile-His¬

11 12 13 IB

and having the sequence to the left of the arrow (↓) being as designated in formula IA above.

Renin cleaves angiotensinogen to produce angiotensin I, which is converted to the potent pressor angiotensin II. A number of angiotensin I converting enzyme inhibitors are known to be useful in the treatment of 'hypertension. Inhibitors of renin are also useful in the treatment of hypertension.

A number of renin-inhibitory peptides have been disclosed. Thus, U.S. Patent 4,424,207; European published applications 45,665; 104,041; and 156,322; and U.S. patent application, Serial No. 825,250, filed 3 February 1986 ; disclose certain peptides with the dipeptide at the 10,11-ρosition containing an isostere bond. A number of statine derivatives stated to be renin inhibitors have been disclosed, see, e.g.,, European published applications 77,028; 81,783; 114,993; 156,319; and 156,321; and U.S. patents 4,478,826; 4,470,971; 4,479,941; and 4,485,099. Terminal disulfide cycles have also been disclosed in renin inhibiting peptides; see, e.g., U.S. patents 4,477,440 and 4,477,441. Aromatic and aliphatic amino acid residues at the 10,11 position of the renin substrate are disclosed in U.S. patents 4,478,827 and 4,455,303. C- terminal amide cycles are disclosed In U.S. patent 4,485,099 and European published applications 156,320 and 156,318. Certain tetrapeptides are disclosed In European publications 111,266 and 77,027. Further, European published application No. 118,223 discloses certain renin inhibiting peptide analogs where the 10-11 peptide link is replaced by a one to four atom carbon or carbon-nitrogen link. Additionally, Holladay et al., in "Synthesis of Hydroxyethylene and Ketomethylene Dipeptide Isosteres", Tetrahedron Letters, Vol. 24, No. 41, pp. 4401-4404, 1983 disclose various intermediates in a process to prepare stereo-direc-ted "ketomethylene" and "hydroxyethylene" dipeptide isosteric functional groups disclosed in the above noted U.S. Patent No. 4,424,207. Evans, et al., J. Org. Chem., 50, 4615 (1985) discloses the synthesis of Hydroxyethylene Dipeptide Isosteres. See also, published European patent application 163,237, which discloses certain renin inhibiting peptides.

Additionally, published European Applications 45,161 and 53,017 disclose amide derivatives useful as inhibitors of angiotensin converting enzymes.

Certain dipeptide and tripeptides are disclosed in U.S. patents 4,514,332; 4,510,085; and 4,548,926 as well as in European published applications 128,762; 152,255; and 181,110. Pepstatin derived renin inhibitors have been disclosed in U.S. patent 4,481,192. Retroinverso bond modifications at positions 10-11 have been disclosed in U.S. patent 4,560,505 and in European published applications 127,234 and 127,235. Derivatives of isosteric bond replacements at positions 10-have been disclosed in European published applications 143,746 and 144,290; and U.S. patent application, Serial No. 904,149, filed 5 September 1986. Isosteric bond modifications at positions 11-12 and 12-13 have been disclosed in European published application 179,352 Certain peptides containing 2-substituted statine analogues have been disclosed in European published application 157,409, Certain peptides containing 3-aminodeoxystatine have been disclosed in European published application 161,588. Certain peptides containing 1-amino- hydroxybutane derivatives at positions 10-11 have been disclosed in European published application 172,346. Certain peptides containing 1- amino-2-hydroxypropane derivatives at positions 10-11 have been disclosed in European published application 172,347. Certain peptides containing N-terminal amide cycles have been disclosed in U.S. patent application, Serial No. 844,716, filed 27 March 1986. Certain peptides containing dihalostatine have been disclosed in PCT application, Serial No. 000,713, filed 7 April 1986. Certain peptides containing C- terminus truncated epoxy or azido or cyano groups or containing a position 10-11 diol and a position 11-12 retro bond have been disclosed in U.S. patent application, Serial No. 945,340, filed 22 December 1986. European published applications 156,322; 114,993; and 118,223; and PCT patent application, Serial No. 002,227, filed 21 November 198 ; U.S. patent application, Serial No. 825,250, filed 3 February 198 ; U.S. patent application, Serial No. 904,149, filed 5 September 198 ; and U.S. patent application, Serial No. 844,716, filed 27 March 198 ; disclose hydroxamic acids or esters at the C-terminus.

E.P. 189,203 discloses new N-dihydroxyalkyl peptide derivatives which are useful as inhibitors of renin for treating hypertension.

E.P. 184,855 discloses new hydroxy substituted-statine peptide derivatives which are useful as inhibitors of renin for treating hypertension.

Derivatives of isosteric bond replacements at positions 10-11 as dihydroxy ethylene isosteres have been disclosed in U.S. patent application, Serial No. 904,149, filed 5 September 1986.

The following references disclose additional substituents at the 10, 11-position: A. Spaltenstein, P. Carpino, F. Miyake and P.B. Hyskins, Tetrahedron Letters, 27:2095 (1986); D.H. Rich and M.S. Bernatowicz, J. Med. Chem., 25:791 (1982); Roger, J. Med. Chem., 28:1062 (1985); D.M. Glick et al., Biochemistry, 21:3746 (1982); D.H. Rich, Biochemistry, 24:3165 (1985); R.L. Johnson, J. Med. Chem., 25:605 (1982); R.L. Johnson and K. Verschovor, J. Med. Chem., 26:1457 (1983); R.L. Johnson, J. Med. Chem., 27:1351 (1984); P.A. Bartlett and W.B. Kezer et al. , J. Am. Chem. Soc., 106:4282 (1984); Peptides: Synthesis. Structure and Function (V.J. Hruby; D.H. Rich, eds.) Proc . 8th American Peptide Sym., Pierce Chemical Company, Rockford, I11., pp. 511-20; 587-590 (1983). INFORMATION DISCLOSURE

Certain peptides having cleavable bonds corresponding to the 10,11-position of the renin substrate and containing cyclopropylamino acids are disclosed in U.S. Patent 4,629,784 (1986) Stammer; PCT Application WO 85/00809 Stammer. Different peptides are shown to have different uses such as food additives, analgetics, CNS regulators, renin inhibitors, and antihypertensive agents. The synthesis of cyclopropylphenylalanine and the preparation of stabilized peptides are disclosed in Kimura, Biochem. and Biophy. Research Commun. 115:112 (1983).

The synthesis of enkephalin analogues containing cyclopropane amino acids is disclosed in CH. Stammer, Pept. Struct. Funct., Proc. Am. Pept. Symp., 287 (1983).

The synthesis of peptides containing cyclopropane amino acids is disclosed in F.H.C. Stewart, Austrian J. of Chemistry, 34:2431 (1981).

Phenylcyclopropane carboxylic acids and esters as hypotensive agents are disclosed in U.S. Patent 4,293,760 (1967) Kaiser; U.S. Patent 3,068,283 (1962) Kaiser; U.S. Patent 3,313,842 (1967) Smith Kline.

An improved preparation of 1-aminocyclopropane-1-carboxylic acid and its use for plant growth regulation are disclosed in U.S. Patent 4,298,760. General syntheses of 2-substituted cyclopropane amino acids are found in the following references: J.E. Baldwin, Tetrahedron Letters, 481, 485 (1985); M. Bernabe, Synthesis, 773 (1985); M. Bernabe, J. Heterocyclic Chem., 20:607 (1983); CH. Stammer, J. Org. Chem., 48:244 (1983); CH. Stammer, Tetrahedron Letters, 24:3839 (1983); CH. Stammer, J. Org. Chem., 47:3270 (1982); M. Bernabe, Eur. J. Med. Chem. Chimica Therapeutics, 14:33 (1979); M. Bernabe, Synthesis, 191 (1977); CH. Stammer, J. Org. Chem., 41:1466 (1976); M. Bernabe, Anales de Quimica, 68:1005 (1972); and W.I. Awad, Tetrahedron, 20:891 (1967).

The synthesis of cyclopropyl thyronine is disclosed in A. Burger and R.A. Pages, J. Med. Chem., 10:435 (1967).

The synthesis of cyclopropyl-3-chloroalanine is disclosed in CH. Stammer, J. Org. Chem., 49:1534 (1984).

The synthesis of 1-ammo-2-(4-imldazolyl)cyclopropane carboxylic acid is disclosed in A. Burger, J. Med. Chem., 9:766 (1966); for related work see A. Burger, J. Med. Chem., 10:435 (1967); C.L. Zirkle, J. Med. Pharm. Chem., 5:1265 (1962); and C Kaiser, Belgian Patent 648,020 (1964). The synthesis of spirocyclopropane amino acids is disclosed in Liebigs Ann. Chem., 611 (1973), U. Schollkopf.

The synthesis of phenethylamine cyclopropane carboxylic acid is disclosed in A. Burger, J. Am. Chem. Soc, 70:2198 (1948); and R.E. Tedeschi, Proc. Soc. Exp. Biol. Med., 102:380 (1959). SUMMARY OF THE INVENTION

The present invention particularly provides:

A renin inhibitory peptide having a non-cleavable transition state insert corresponding to the 10, 11-position of a renin substrate (angiotensinogen) and having: (a) at least one cyclopropyl amino acid of the formula XL_2b in place of an amino acid residue corresponding to position 6, 8, 9, 12, 13 or 14 of the renin substrate: and/or

(b) a cycloalkyl amino acid non-cleavable transition state insert of the formula XL_2c in place of an amino acid residue corresponding to position 10 and 11 of the renin substrate; wherein R₅₀ is

(a) hydrogen,

(b) C₁-C₅alkyl,

(c) -(CH₂)_p-aryl, (d) -(CH₂)_p-Het,

(e) -(CH₂)p-C₃-C₇cycloalkyl, or

(f) 1- or 2-adamantyl; wherein R₆₀ and R₆₁ are the same or different and are

(a) hydrogen, (b) C₁-C₅alkyl ,

(c) - (CH₂ )_p- aryl ,

(d) - (CH₂)_p -Het ,

(e) - (CH₂)_p -C₃-C₇cycloalkyl, or

( f) 1 - or 2 - adamantyl ; or wherein R₆₀ and R₆₁ taken together form a spirocycle of the formula II wherein R₇₀ is

(a) -CHR₈₀ , (b) -O-,

(c) -S-,

(d) -SO-,

(e) -SO₂-, or (f) -NR₈₁; wherein R₈₀ and R₈₁ are the same or different and are

(a) hydrogen,

(b) C₁-C₅alkyl,

(c) -(CH₂)_p-aryl, (d) -(CH₂)_p-Het,

(e) -(CH₂)_p-C₃-C₇ cycloalkyl, or

(f) 1- or 2-adamantyl; wherein R₉₀ is

(a) hydrogen, or (b) C₁-C₅alkyl; wherein R₁₀₀ is

(a) hydrogen,

(b) C₁-C₅alkyl,

(c) aryl, (d) C₃-C₇cycloalkyl,

(e) -Het,

(f) C₁-C₃alkoxy, or

(g) C₁-C₃alkylthio; wherein R₁₀₀ is (a) hydrogen,

(b) C₁-C₅alkyl,

(c) -(CH₂)_p-aryl, or

(d) halogen; wherein R₁₁₁ is (a) hydrogen,

(b) C₁-C₅alkyl,

(c) -(CH₂)_p-aryl, or

(d) halogen; wherein p is zero to two, inclusive; wherein r is zero to three, inclusive; wherein t is zero to three, Inclusive; wherein aryl is phenyl or naphthyl substituted by zero to 3 of the following: (a) C₁-C₃alkyl,

(b) hydroxy,

(c) C₁-C₃alkoxy,

(d ) halo ,

(e) amino ,

(f ) mono- or di-C₁-C₃alkylamino,

(g ) -CHO,

(b) -COOH ,

(i) COOR₂₆ ,

(j) CONHR₂₆,

(k ) nitro,

(l) mercapto,

(m ) C₁-C₃alkylthio,

(n ) C₁-C₃alkylsulfinyl,

(o) C₁-C₃alkylsulfonyl,

(p) -N(R₄)-C₁-C₃alkylsulfonyl,

(q) SO₃H,

(r) SO₂NH₂,

(s) -CN, or

( t) -CH₂NH₂; wherein -Het is a 5- or 6-membered saturated or unsaturated ring containing from one to three heteroatoms selected from the group consisting of nitrogen, oxygen, and sulfur; and including any bicyclic group in which any of the above heterocyclic rings is fused to a benzene ring, which heterocyclic moiety is substituted with zero to 3 of the following:

(i) C₁-C₆alkyl, (ii) hydroxy, (iii) trifluoromethyl, (iv) C₁-C₄alkoxy,

(v) halo, (vi) aryl, (vii) aryl C₁-C₄alkyl-, (viii) amino,

(ix) mono- or di- (C₁-Chalkyl)amino, and (x) C₁-C₅alkanoyl; wherein R₄ at each occurrence is the same or different and is (a) hydrogen, (b) C₁-C₅alkyl,

(c) -(CH₂)_p-aryl,

(d) -(CH₂)_p-Het,

(e) -(CH₂)p-C₃-C₇ cycloalkyl, or (f) 1- or 2-adamantyl; wherein R₂₆ is

(a) hydrogen,

(b) C₁-C₃alkyl, or

(c) phenyl-C₁-C₃alkyl. In a renin inhibitory peptide having a non-cleavable transition state insert corresponding to the 10 , 11-position of a renin substrate (angiotensinogen) , the improvement which comprises inclusion in the renin inhibitory peptide of:

(a) at least one cyclopropyl amino acid of the formula XL_2b in place of an amino acid residue corresponding to position 6 , 8 , 9 , 12 ,

13 or 14 of the renin substrate ; and/or

(b) a cycloalkyl amino acid non-cleavable transition state insert of the formula XL_2c in place of an amino acid residue corresponding to position 10 and 11 of the renin substrate ; wherein R₅₀ is

(a) hydrogen,

(b) C₁-C₅alkyl,

(c) -(CH₂)_p-aryl,

(d) -(CH₂)_p-Het, (e) -(CH₂)p-C₃-C₇ cycloalkyl, or

(f) 1- or 2-adamantyl; wherein R₆₀ and R₆₁ are the same or different and are

(a) hydrogen,

(b) C₁-C₅alkyl, (c) -(CH₂)_p-aryl,

(d) -(CH₂)_p-Het,

(e) -(CH₂)p-C₃-C₇cycloalkyl, or

(f) 1- or 2-adamantyl; or wherein R₆₀ and R₆₁ taken together form a spirocycle of the formul II wherein R₇₀ is

(a) -CHR₈₀,

(b) -O-, (c) -S-,

(d) -SO-,

(e) -SO₂-, or

(f) -NR₈₁; wherein R₈₀ and R₈₁ are the same or different and are

(a) hydrogen,

(b) C₁-C₅alkyl,

(c) -(CH₂)_p-aryl,

(d) -(CH₂)_p-Het, (e) -(CH₂)_p-C₃-C₇ cycloalkyl, or

(f) 1- or 2-adamantyl; wherein R₉₀ is

(a) hydrogen, or

(b) Cx-Csalkyl; wherein R₁₀₀ ^is

(a) hydrogen,

(b) C₁- C₅alkyl ,

(c) aryl ,

(d) C₃ -C₇cycloalkyl , (e) -Het ,

(f) C₁-C₃alkoxy , or

(g) C₁-C₃alkylthio ; wherein R₁₁₀ is

(a) hydrogen , (b) C₁-C₅alkyl ,

(c) -(CH₂)p-aryl, or

(d) halogen; wherein R₁₁₁ is

(a) hydrogen, (b) C₁-C₅alkyl,

(c) -(CH₂)_p-aryl, or

(d) halogen; wherein p is zero to two, inclusive; wherein r is zero to three, inclusive; wherein t is zero to three, inclusive; wherein aryl is phenyl or naphthyl substituted by zero to three the following:

(a) C₁-C₃alkyl, (b) hydroxy ,

(c) C₁ -C₃alkoxy,

(d) halo ,

(e) amino , (f) mono- or di-C₁-C₃alkylamino,

(g) -CHO, (h) -COOH, (i) COOR₂₆, (j) CONHR₂₆ , (k) nitro,

(l) mercapto, (m) C₁-C₃alkylthio, (n) C₁-C₃alkylsulfinyl, (o) C₁-C₃alkylsulfonyl, (p) -N(R₄)-C₁-C₃alkylsulfonyl,

(q) SO₃H, (r) SO₂NH₂, (s) -CN, or (t) -CH₂NH₂; wherein -Het is a 5- or 6-membered saturated or unsaturated ring containing from one to three heteroatoms selected from the group consisting of nitrogen, oxygen, and sulfur; and including any bicyclic group in which any of the above heterocyclic rings is fused to a benzene ring, which heterocyclic moiety is substituted with zero to three of the following:

(i) C₁-C₆alkyl,

(ii) hydroxy,

(iii) trifluoromethyl,

(iv) C₁-C₄alkoxy,

(v) halo,

(vi) aryl,

(vii) aryl C₁-C₄alkyl-,

(viii) amino,

(ix) mono- or di-(C₁-C₄alkyl)amino, and

(x) C₁-C₅alkanoyl ; wherein R₄ at each occurrence is the same or different and is

(a) hydrogen,

(b) C₁-C₅alkyl, (c) -(CH₂)_p-aryl,

(d) -(CH₂)_p-Het,

(e) -(CH₂)p-C₃-C₇ cycloalkyl, or

(f) 1- or 2-adamantyl; wherein R₂₆ is

(a) hydrogen,

(b) C₁-C₃alkyl, or

(c) phenyl-C₁-C₃alkyl.

Examples of renin inhibitors of the present invention of formula I are: wherein X is

(a) hydrogen,

(b) C₁-C₅alkyl,

(c) R₅-O-CH₂-C(O)-,

(d) R₅-CH₂-O-C(O)-, (e) R₅-O-C(O)-,

(f) R₅-(CH₂)_n-C(O)-,

(g) R₄N(R₄)-(CH₂)_n-C(O), (h) R₅-SO₂-(CH₂)_q-C(O)-,

(i) R₅-SO₂-(CH₂)_q-O-C(O)-, or (j) R₆-(CH₂)_i-C(O)-; wherein A₆ is absent or a divalent moiety of the formula XL₁, XL₂ XL_2a, or XL_2b; wherein B₇ is absent or a divalent moiety of the formula XL_b; wherein C₈ is absent or a divalent moiety of the formula XL₁, XL₂, XL_2a, or XL_2b; wherein D₉ is absent or a divalent moiety of the formula XL₃, XL_2a, or XL_2b; or wherein C_8-D₉ is XL₇ , XL_7a, or when X, A₆ and B₇ are absent,

^XL7b; wherein E₁₀-F₁₁ is a divalent moiety of the formula XL₆ , XL_6a,

XL_6b, XL_6c, XL_6d, XL_6e, XL_6f, or XL_6g; wherein * indicates an asymmetric center which is either in the R or S configuration; wherein G₁₂ is absent or a divalent moiety of the formula XL_4, XL_4a, or XL_2b; wherein H₁₃ is absent or a divalent moiety of the formula XL₄ or

^XL2b ; wherein I₁₄ is absent or a divalent moiety of the formula XL₅ or XL_2b ; wherein 2 is

(a) -O-R₁₀,

(b) -N(R₄)R₁₄, or

(c) C₄-C₈cyclic amino; wherein R is

(a) isopropyl,

(b) isobutyl,

(c) phenylmethyl, or

(d) C₃-C₇cycloalkyl; wherein R₁ is

(a) hydrogen,

(b) C₁-C₅alkyl,

(c) aryl,

(d) C₃-C₇cycloalkyl,

(e) -Het,

(f) C₁-C₃alkoxy, or (g) C₁-C3alkylthio; wherein R₂ is

(a) hydrogen, or

(b) -CH(R₃)R₄; wherein R₃ is

(a) hydrogen,

(b) hydroxy,

(c) C₁-C5alkyl,

(d) C₃-C₇cycloalkyl,

(e) aryl,

(f) -Het,

(g) C₁-C₃alkoxy, or (h) C₁-C₃alkylthio; wherein R₄ at each occurrence is the same or different and is

(a) hydrogen,

(b) C₁-C₅alkyl ,

(c) - (CH₂)p-aryl, (d) - (CH₂)p-Het ,

(e) - (CH₂)p-C₃-C₇cycloalkyl , or

(f) 1- or 2-adamantyl ; wherein R₅ is (a) C₁ - C₆alkyl ,

(b) C₃-C₇cycloalkyl,

(c) aryl,

(d) -Het, or

(e) 5-oxo-2-pyrrolidinyl; wherein R₆ is

(a) hydrogen,

(b) C₁-C₅alkyl,

(c) -(CH₂)_p-aryl,

(d) -(CH₂)_p-Het,

(e) -(CH₂)_p-C₃-C₇cycloalkyl, or

(f) 1- or 2-adamantyl; wherein R₇ is

(a) hydrogen,

(b) C₁-C₅alkyl,

(c) hydroxy,

(d) amino C₁-C₄alkyl-,

(e) guanidinyl C₁-C₃alkyl-,

(f) aryl,

(g) -Het,

(h) methylthio,

(i) -(CH₂)_p-C₃-C₇cycloalkyl, or

(j) amino ; wherein R₈ is

(a) hydrogen,

(b) C₁-C₅alkyl,

(c) hydroxy,

(d) aryl,

(e) -Het,

(f) guanidinyl C₁-C₃alkyl-, or

(g) -(CH₂)_p-C₃-C₇cycloalkyl; wherein R₉ is

(a) hydrogen,

(b) hydroxy,

(c) amino C₁-C₄alkyl-, or

(d) guanidinyl C₁-C₃alkyl-; wherein R₁₀ is

(a) hydrogen, (b) C₁-C₅alkyl,

(c) -(CH₂)_nR₁₆,

(d) -(CH₂)_nR₁₇,

(e) C₃-C₇cycloalkyl,

(f) a pharmaceutically acceptable cation,

(g) -CH(R₂₅)-CH₂-R₁₅, or (h) -CH₂-CH(R₁₂)-R₁₅; wherein R₁₁ is -R or -R₂; wherein R₁₂ is -(CH₂)_n-R₁₃; wherein R₁₃ is

(a) aryl,

(b) amino,

(c) mono-, di or tri-C₁-C₃alkylamlno,

(d) -Het,

(e) C₁-C₅alkyl

(f) C₃-C₇cycloalkyl,

(g) C₂-C₅alkenyl, (h) C₃-C₇cycloalkenyl,

(i) hydroxy,

(j) C₁-C₃alkoxy, (k) C₁-C₃alkanoyloxy,

(l) mercapto,

(m) C₁-C₃alkylthio, (n) -COOH,

(o) -CO-O-C₁-C₆alkyl,

(p) -CO-O-CH₂-(C₁-C₃alkyl)-N(C₁-C₃alkyl)₂,

(q) -CO-NR₂₂R₂₆;

(r) C₄-C₇cyclic amino,

(s) C₄-C₇cycloalkylamino,

(t) guanidyl,

(u) cyano,

(v) N-cyanoguanidyl,

(w) cyanoamino,

(x) (hydroxy C₂-C₄alkyl)amino, or

(y) di- (hydroxyC₂-C₄alkyl)amino; wherein R₁₄ is

(a) hydrogen,

(b) C₁-C₁₀alkyl, (c) -(CH₂)_n-R₁₈,

(d) -(CH₂)_n-R₁₉,

(e) -CH(R₂₅)-CH₂-R₁₅,

(f) -CH₂-CH(R₁₂)-R₁₅, (g) (hydroxy C₁-C₈alkyl), or

(h) (C₁-C₃alkoxy)C₁-C₈alkyl; wherein R₁₅ is

(a) hydroxy,

(b) C₃-C₇cycloalkyl, (c) aryl,

(d) amino,

(e) mono-, di- , or tri- C₁-C₃alkylamino,

(f) mono- or di- (hydroxy C2 -C₄alkyl) amino,

(g) -Het, (h) C₁-C₃alkoxy-,

(i) C₁-C₃alkanoyloxy-,

( j ) mercapto,

(k) C₁-C₃alkylthio-,

(l) C₁-C₅alkyl, (m) C₄-C₇cyclic amino,

(n) C₄-C₇cycloalkylamino,

(o) C₁-C alkenyloxy,

(p) C₃-C₇cycloalkenyl; wherein R₁₆ is (a) aryl,

(b) amino,

(c) mono- or di- (C₁-C₃alkyl)amino,

(d) hydroxy,

(e) C₃-C₇cycloalkyl, (f) C₄-C₇cyclic amino, or

(g) C₁-C₃alkanoyloxy; wherein R₁₇ is

(a) -Het,

(b) C₁-C₅alkenyl, (c) C₃-C₇cycloalkenyl,

(d) C₁-C₃alkoxy,

(e) mercapto,

(f) C₁-C₃alkylthio, (g) -COOH,

(h) -CO-O-C₁-C₆alkyl,

(i) -CO-O-CH₂-(C₁-C₃alkyl)-N(C₁-C3alkyl)₂,

(j) -CO-NR₂₂R₂₆, (k) tri-C₁-C₃alkylamino,

(l) guanidyl,

(m) cyano,

(n) N-cyanoguanidyl,

(o) (hydroxy C₂-C₄alkyl)amino, (p) di- (hydroxy C₂-C₄alkyl)amino, or

(q) cyanoamino; wherein R₁₈ is

(a) amino,

(b) mono-, or di- (C₁-C₃alkyl)amino, (c) C₄-C₇cyclic amino; or

(d) C₄-C₇cycloalkylamlno; wherein R₁₉ is (a) aryl,

(b) -Het,

(c) tri-C ₁-C₃alkylamino ,

(d) C₃-C₇cycloalkyl,

(e) C₁-C₅alkenyl,

(f) C₃-C₇cycloalkenyl,

(g) hydroxy, (h) C₁-C₃alkoxy,

(i) C₁-C₃alkanoyloxy,

(j) mercapto, (k) C₁-C₃alkylthio,

(l) -COOH, (m) -CO-O-C₁-C₆alkyl,

(n) -CO-O-CH₂-(C₁-C₃alkyl)-N(C₁-C₃alkyl)₂,

(o) -CO-NR₂₂R₂₆,

(p) guanidyl,

(q) cyano,

(r) N-cyanoguanidyl,

(s) cyanoamino,

(t) (hydroxy C₂-C₄alkyl)amino, (u) di- (hydroxy C₂-C₄alkyl)amino; or (v) -SO₃H; wherein R₂₀ is

(a) hydrogen,

(b) C₁-C₅alkyl, or

(c) aryl-C₁-C₅alkyl; wherein R₂₁ is

(a) -NH₂, or

(b) -OH; wherein R₂₂ is

(a) hydrogen, or

(b) C₁-C₃alkyl; wherein R₂₃ is

(a) -(CH₂)_n-OH,

(b) -(CH₂)_n-NH₂,

(c) aryl, or

(d) C₁-C₃alkyl; wherein R₂₄ is

(a) -R₁,

(b) -(CH₂)_n-OH, or

(c) -(CH₂)_n-NH₂; wherein R₂₅ is

(a) hydrogen,

(b) C₁-C₃alkyl, or

(c) phenyl -C₁-C₃alkyl; wherein R₂₆ is

(a) hydrogen,

(b) C₁-C₃alkyl, or

(c) phenyl -C₁-C₃alkyl; wherein R₅₀ is

(a) hydrogen,

(b) C₁-C₅alkyl,

(c) -(CH₂)_p-aryl,

(d) -(CH₂)_p-Het,

(e) -(CH₂)p-C₃-C₇cycloalkyl, or

(f) 1- or 2-adamantyl; wherein R₆₀ and R₆₁ are the same or different and are

(a) hydrogen,

(b) C₁-C₅alkyl, (c) -(CH₂)_p-aryl,

(d) -(CH₂)_p-Het,

(e) -(CH₂)_p-C₃-C₇cycloalkyl, or

(f) 1- or 2-adamantyl; erein R₆₀ and R₆₁ taken together form a spirocycle of the formula

wherein R₇₀ is

(a) -CHR₈₀,

(b) -O-, (c) -S-,

(d) -SO-,

(e) -SO₂-, or

(f) -NR₈₁; wherein R₈₀ and R₈₁ are the same or different and are (a) hydrogen,

(b) C₁-C₅alky!,

(c) -(CH₂)_p-aryl,

(d) -(CH₂)_p-Het,

(e) -(CH₂)_p-C₃-C₇cycloalkyl, or (f) 1- or 2-adamantyl; wherein R₉₀ is

(a) hydrogen, or

(b) C₁-C₅alkyl; wherein R100 is (a) hydrogen,

(b) C₁-C₅alkyl,

(c) aryl,

(d) C₃-C₇cycloalkyl,

(e) -Het, (f) C₁-C₃alkoxy, or

(g) C₁-C₃alkylthio; wherein R₁₁₀ is

(a) hydrogen,

(b) C₁-C₅alkyl, (c) - (CH₂)_p-aryl , or

(d) halogen; wherein R_{1 11} is

(a) hydrogen, (b) C₁-C₅alkyl,

(c) -(CH₂)_p-aryl, or (d) halogen; wherein i is zero to five, inclusive; wherein m is one or two; wherein for each occurrence n is independently an integer of zero to five, inclusive; wherein p is zero to two, inclusive; wherein q is one to five, inclusive; wherein r is zero to three, inclusive; wherein t is zero to three, inclusive; wherein Q is

(a) -CH₂-,

(b) -CH(OH)-, (c) -O-, or

(d) -S-; and wherein M is

(a) -CO-, or

(b) -CH₂-; wherein aryl is phenyl or naphthyl substituted by zero to three of the following:

(a) C₁-C₃alkyl,

(b) hydroxy,

(c) C₁ -C₃alkoxy, (d) halo,

(e) amino,

(f) mono- or di-C₁-C₃alkylamino,

(g) -CHO, (h) -COOH, (i) COOR₂₆,

(j) CONHR₂₆ ,

(k) nitro,

(l) mercapto,

(m) C₁-C₃alkylthio, (n) C₁-C₃alkylsulfinyl,

(o) C₁-C₃alkylsulfonyl,

(p) -N(R₄)-C₁-C₃alkylsulfonyl,

(q) SO₃H, (r) SO₂NH₂, (s) -CN, or (t) -CH₂NH₂; wherein -Het is a 5- or 6-membered saturated or unsaturated ring containing from one to three heteroatoms selected from the group consisting of nitrogen, oxygen, and sulfur; and including any bicyclic group In which any of the above heterocyclic rings is fused to a benzene ring, which heterocyclic moiety is substituted with zero to three of the following: (i) C₁-C₆alkyl,

(ii) hydroxy, (iii) trifluoromethyl, (iv) C₁-C₄alkoxy, (v) halo, (vi) aryl,

(vil) aryl C₁-C₄alkyl-, (viii) amino,

(ix) mono- or di- (C₁-C₄alkyl)amino, and (x) C₁-C₅alkanoyl; with the overall provisos that:

(1) at least one cyclopropyl amino acid of the formula XL_2b or cycloalkyl amino acid non-cleavable transition state insert of the formula XL_2c is present;

(2) R₁₈ or R_1g is hydroxy, mercapto, or amino, or a mono-substituted nitrogen containing group bonded through the nitrogen only when n is not one;

(3) R₁₂ is -(CH₂)_n-R₁₃ and n is zero and both R₁₃ and R₁₅ are oxygen-, nitrogen-, or sulfur-containing substituents bonded through the hetero atom, only when the hetero atom is not also bonded to hydrogen;

(4) R₁₇ or R₁₉ is -COOH only when n for that moiety is other than zero;

(5) R₁₆ or R₁₇ is an amino-containing substituent, hydroxy, mercapto, or -Het bonded through the hetero atom only when n for that substituent is an integer from two to five, inclusive;

(6) when R₁₂ is - (CH₂)n-R₁₃ and n is zero, then R₁₃ and R₁₅ cannot both be -COOH; and

(7) R₁₇ or R₁₉ is -Het, only when -Het is other than cyclic amino; or a carboxy-, amino-, or other reactive group-protected form thereof; or a pharmaceutically acceptable acid addition salt thereof. In formula XL_2c, both R₁₁₀ and R₁₁₁ are attached to the same carbon atom of the cycloalkyl ring.

By "renin inhibitory peptide" is meant a compound capable of inhibiting the renin enzyme in mammalian metabolism and having thre or more amino acid residues linked by peptidic or pseudo-peptidic bonds. By "a non-cleavable transition state insert" is meant a transition state insert which is not cleavable by a hydrolytic enzyme in mammalian metabolism. A variety of such transition state inserts, corresponding to the 10,11-position of the renin substrate, are known in the art including those disclosed in the following references, which are hereby incorporated by reference:

U.S. Patent 4,424,207 (Szelke); European Patent 104041A (Szelke); European Patent Application 144.290A (Ciba Geigy AG) ; European Patent 0,156,322 (Merck); European Patent 161-588A (Merck); European Patent 0,172,347 (Abbott); European Patent 172-346-A (Abbott); European Patent 156-318 (Merck); European Patent 157-409 (Merck); European Patent 152-255 (Sankyo); and U.S. Patent 4,548,926 (Sankyo); and

U.S. patent application, Serial No. 904,149, filed 5 September

1986; U.S. patent application, Serial No. 844,716, filed 27 March 1986;

PCT application, Serial No. 000,713, filed 7 April 1986; U.S. patent application, Serial No. 945,340, filed 22 December 1986; and U.S. patent application, Serial No. 825,250, filed 3 February 1986; and

A. Spaltenstein, P. Carpino, F. Miyake and P.B. Hyskins, Tetrahedron Letters, 27:2095 (1986); D.H. Rich and M.S. Bernatowicz, J. Med. Chem., 25:791 (1982); Roger, J. Med. Chem., 28:1062 (1985); D.M. Glick et al., Biochemistry, 21:3746 (1982); D.H. Rich, Biochemistry, 24:3165 (1985); R.L. Johnson, J. Med. Chem., 25:605 (1982); R.L. Johnson and K. Verschovor, J. Med. Chem., 26:1457 (1983); R.L. Johnson, J. Med. Chem., 27:1351 (1984); P.A. Bartlett et al., J. Am. Chem. Soc, 106:4282 (1984); arid Peptides: Synthesis, Structure and Function (V.J. Hruby; D.H. Rich, eds.) Proc. 8th American Peptide Sym., Pierce Chemical Company, Rockford, 111., pp. 511-20; 587-590 (1983).

As is apparent to those of ordinary skill in the art, the renin inhibitory peptides of the present invention can occur in several isomeric forms, depending on the configuration around the asymmetric carbon atoms. All such isomeric forms are included within the scope of the present invention. The E isomer of the cyclopropyl amino acids is preferred. The trans isomer of the cycloalkyl transition state inserts is preferred. Preferably, the stereochemistry of the other amino acids corresponds to that of the naturally-occurring amino acids.

Renin inhibitory peptides commonly have protecting groups at the

N-terminus and the C-terminus. These protecting groups are known In the polypeptide art. Examples of these protecting groups are given below. Any of these protecting groups are suitable for the renin inhibitory peptides of the present invention.

Furthermore, the cyclopropyl amino acid of the formula XL_2b of the present Invention may occur at the N-terminus of the renin inhibitory peptide and, as such, will, when coupled with a suitable protecting group, assume the ending position.

These compounds are shown in relation to the human renin substrate as follows :

6 7 8 9 10 11 12 13 -His Pro Phe His Leu Val He His- X A₆ B₇ C₈ D₉ E₁₀ F₁₁ G₁₂ H₁₃ I₁₄ Z,

The present invention provides peptide inhibitors of renin which contain at least one cyclopropane amino acid and have transition state inserts.

Examples of pharmaceutically acceptable acid addition salts include: acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, citrate, camphorate, camphorsulfonate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, fumarate, glucoheptanoate, glycerophosphate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydrσiodide, 2-hydroxyethanesulfonate, lactate, maleate, methanesulfonate, 2-naphthalenesulfonate, nlcotinate, oxalate, palmoate, pectinate, persulfate, 3-phenylpropionate, plcrate, pivalate, propionate, succinate, tartrate, thiocyanate, tosylate, and undecanoate.

The carbon atom ccntent of various hydrocarbon-containing moieties is indicated by a prefix designating the minimum and maximum number of carbon atoms in the moiety, i.e., the prefix (G_i-C_j) indicates a moiety of the integer "i" to the integer "j" carbon atoms, inclusive. Thus

(C₁-C₄)alkyl refers to alkyl of one to 4 carbon atoms, inclusive, or methyl, ethyl, propyl, butyl, and isomeric forms thereof. C₄-C₇cyclic amino indicates a monocyclic group containing one nitrogen and 4 to 7 carbon atoms.

Examples of (C₃-C₁₀)cycloallcyl which include alkyl-substituted cycloalkyl containing a total of up to 10 total carbon atoms, are cyclopropyl, 2-methylcyclopropyl, 2,2-dimethylcycloρropyl, 2,3-diethylcyclopropyl, 2-butylcycloproρyl, cyclobutyl, 2-methylcyclobutyl, 3- propylcyclobutyl, cyclopentyl, 2,2-dimethylcyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl and isomeric forms thereof.

Examples of aryl include phenyl, naphthyl, (o-, m-, p-)tolyl, (o-, m-, p-)ethylphenyl, 2-ethyl- tolyl, 4-ethyl-o-tolyl, 5-ethyl-m-tolyl, (o-, m-, or p-)propylphenyl, 2-propyl- (o-, m-, or p-) tolyl, 4-isopropyl-2,6-xylyl, 3-propyl-4-ethylphenyl, (2,3,4- 2,3,6-, or 2,4,5-)- trimethylphenyl, (o-, m-, or p-)fluorophenyl, (o-, m-, or p-trifluoromethyl)phenyl, 4-fluoro-2,5-xylyl, (2,4-, 2,5-, 2,6-, 3,4-, or 3,5-)difluorophenyl, (o-, m-, or p-)chlorophenyl, 2-chloro-p- tolyl, (3-, 4- 5- or 6-)chloro-o-tolyl, 4-chloro-2-propylphenyl, 2-isopropyl-4-chlorophenyl , 4- chloro- 3 - fluorophenyl , ( 3 - or 4- ) chloro - 2 - f luorophenyl , (o-, m-, or p-)trifluoro-methylphenyl, (o-, m-, or p-)ethoxyphenyl, (4- or 5- )chloro-2 -methoxy-phenyl, and 2,4-dichloro(5- or 6-)methylphenyl, and the like.

Examples of -Het include: 2-, 3-, or 4-pyridyl, imidazolyl, indolyl, Nⁱⁿ-formyl-indolyl, Nⁱⁿ-C₁-C₅alkyl-C(O)-indolyl, [1,2,4]- triazolyl, 2-, 4-, or 5-pyrimidinyl, 2- or 3-thienyl, piperidinyl, pyrryl, pyrrolinyl, pyrrolidinyl, pyrazolyl, pyrazolinyl, pyrazolidi- nyl, imidazolinyl, imidazolidinyl, pyrazinyl, piperazinyl, pyridazinyl, oxazolyl, oxazolidinyl, isoxazolyl, isoxazolidinyl, morpholinyl, thiazolyl, thiazolidinyl, isothiazolyl, isothiazolidinyl, quinolinyl, isoquinolinyl, benzimidazolyl, benzothiazolyl, benzoxazolyl, furyl, thienyl, and benzothienyl. Each of these moieties may be substituted as noted above.

As would be generally recognized by those skilled in the art of organic chemistry, a heterocycle as defined herein for -Het would not be bonded through oxygen or sulfur or through nitrogen which is within a ring and part of a double bond.

Halo is halogen (fluoro, chloro, bromo, or iodo) or trifluoromethyl. Examples of pharmaceutically acceptable cations include: pharmacologically acceptable metal cations, ammonium, amine cations, or quaternary ammonium cations. Especially preferred metal cations are those derived from the alkali metals, e.g., lithium, sodium, and potassium, and from the alkaline earth metals, e.g., magnesium and calcium, although cationlc forms of other metals, e.g., aluminum, zinc, and iron are also within the scope of this invention. Pharmacologically acceptable amine cations are those derived from primary, secondary, or tertiary amines. The novel peptides herein contain both natural and synthetic amino acid residues. These residues are depicted using standard amino acid abbreviations (see, e.g., Eur. J. Biochem., 138, 9 (1984)) unless otherwise indicated.

In addition to the treatment of warm-blooded animals such as mice, rats, horses, dogs, cats, etc., the compounds of the Invention are effective In the treatment of humans.

The renin inhibitors of this Invention are useful for treating any medical condition for which it is beneficial to reduce the levels of active circulating renin. Examples of such conditions include renin-associated hypertension and hyperaldosteronism, hypertension, hypertension under treatment with another antihypertensive and/or a diuretic agent, congestive heart failure, angina, and post-myocardial infarction. The renin-anglotension system may play a role in maintenance of intracellular homeostasis: see Clinical and Experimental Hypertension, 86, 1739-1742 (1984) at page 1740 under Discussion.

Further, the renin inhibitors of this invention may be useful in the treatment of cerebrovascular disorders and disorders of intracellular homeotasis. The possible role of the renin-angiotensin system in the maintenance of intracellular homeostasis is disclosed in Clinical and Experimental Hypertension, 86:1739-1742 (1984). Additionally, the renin inhibitors of this invention potentiate the antithrombotic activity of a thromboxane antagonist (U.S. patent 4,558,037). The antihypertensive effect of the renin inhibitors of this invention are potentiated by combination with a thromboxane synthetase inhibitor. The compounds of the present invention are preferably orally administered to humans to effect renin inhibition for the purpose of favorably affecting blood pressure. For this purpose, the compounds are administered from 0.1 mg to 1000 mg per kg per dose, administered from 1 to 4 times daily. The compounds of the present invention are preferably orally administered in the form of pharmacologically acceptable acid addition salts. Preferred pharmacologically acceptable salts for oral administration include the citrate and aspartate salts , although any pharmacologically acceptable salt is useful in this invention, including those listed above. These salts may be in hydrated or solvated form.

Other routes of administration include parenteral, by inhalation spray, or rectally in dosage unit formulations containing conventional non-toxic pharmaceutically acceptable carriers, adjuvants and vehicles. The term parenteral as used herein includes subcutaneous injections, intravenous, intramuscular, intrasternal injection or infusion techniques.

The pharmaceutical compositions may be in the form of a sterile injectable preparation, for example as a sterile injectable aqueous or oleagenous suspension. This suspension may be formulated according to the known art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic 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. 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 injectibles.

Equivalent dosages for such other routes of administration are thus employed. The exact dose depends on the age, weight, and condition of the patient and on the frequency and route of administration. Such variations are within the skill of the practitioner or can readily be determined.

The compounds of the present invention may be in the form of pharmaceutically acceptable salts both those which can be produced from the free bases by methods well known in the art and those with which acids have pharmacologically acceptable conjugate bases.

Conventional forms and means for administering renin-inhibiting compounds may be employed and are described, e.g., in U.S. Patent No. 4,424,207 which is incorporated by reference herein. Likewise, the amounts disclosed In the U.S. Patent No. 4,424,207 are examples applicable to the compounds of the present invention.

The renin-inhibiting compounds of this invention may be administered in combination with other agents used in antihypertensive therapy such as diuretics, or and/or β-adrenergic blocking agents, CNS -acting agents, adrenergic neuron blocking agents, vasodilators, angiotensin I converting enzyme inhibitors, and the like as described, for example, in published European patent application 156 318.

For example, the compounds of this invention can be given in combination with such compounds or salts or other derivative forms thereof as :

Diuretics: acetazolamide; amiloride; bendroflumethiazide; benzthiazide; bumetanide; chlorothiazide; chlorthalidone; cyclothiazide; ethacrynic acid; furosemide; hydrochlorothiazide; hydroflumethlazide; indacrinone (racemic mixture, or as either the (+) or (-) enantiomer alone, or a manipulated ratio, e.g., 9:1 of said enantiomers, respectively); metolazone; methyclothiazide; muzolimine; polythiazide; quinethazone; sodium ethacrynate; sodium nitroprusside; spironolactone; ticrynaten; trimaterene; trichlormethlazide; α-Adrenergic Blocking Agents: dibenamine; phentolamine; phenoxybenzamine; prazosin; tolazoline; β-Adrenergic Blocking Agents: atenolol; metoprolol; nadolol; propranolol; timolol; ((±)-2-[3-(tert-butylamino)-2-hydroxypropoxy]-2-furananilide) (ancarolol);

(2-acetyl-7-(2-hydroxy-3-isopropylaminopropoxy)benzofuran HCl) (befunolol);

((±)-1-(isopropylamino)-3-(p-(2-cyclopropylmethoxyethyl)-phenoxy)-2-propranol HCl) (betaxolol); (1-[ (3,4-dimethoxyphenethyl) amino]-3-(m-tolyloxy)-2-propanol HCl) (bevantolol);

(( (±)-1 -(4-( (2-isopropoxyethoxy)methyl)phenoxy)-3-isopropylamino-2-propanol)fumarate) (blsoprolol); (4-(2-hydroxy-3-[4-(phenoxymethyl)-piperidino]-pfopoxy)-indole); (carbazolyl-4-oxy-5,2-(2-methoxyphenoxy)-ethylamino-2-propanol);

(1-((1,1-dimethylethyl)amino)-3-((2-methyl 'H-indol-4-yl)oxy)-2-propanol benzoate) (bopindolol);

(1-(2-exobicyclo[2.2.1]-hept-2-ylphenoxy)-3-[(1-mathylethyl)-amino]-2- propanol HCl) (bornaprolol);

( o -[2-hydroxy-3-[(2-indol-3-yl-1,1-dimethylethyl)-amino] propoxy] benzonitrile HCl) (bucindolol);

(α- [ (tert.butylamino)methyl]-7-ethyl-2-benzofuranmethanol) (bufur-alo- 1);

(3-[3-acetyl-4-[3-(tert.butylamino)-2-hydroxypropyl]-phenyl]-1,1- diethylurea HCl) (celiprolol);

((±)-2-[2-[3-[(1,1-dimethylethyl) amino]-2-hydroxypropoxy]phenoxy]-N- methylacetamide HCl) (cetamolol); (2-benzimidazolyl-phenyl(2-isopropylaminopropanol));

((±)-3'-acetyl-4'-(2-hydroxy-3-isopropylaminopropoxy)-acetanilide HCl)

(diacetolol);

(methyl-4-[2-hydroxy-3-[(1-methylethyl)aminopropoxyl]]-benzene-propanoate HCl) (esmolol); (erythro-DL-1-(7-methylindan-4-yloxy)-3-isopropylaminobutan-2-ol);

(1- (tert. butylamino)-3-[0-(2-propynyloxy)phenoxy]-2-propanol (pargolol);

(1- (tert. butylamino)-3-[o-(6-hydrazino-3-pyridazinyl)phenoxy]-2-propanol diHCl) (prizidilol); ( (-)-2-hydroxy-5-[(R)-1-hydroxy-2-[(R)-(1-methyl-3-phenylpropyl)-amino] ethyl]benzamide);

(4 -hydroxy-9-[2-hydroxy-3-(isopropylamino)-propoxy]-7-methyl-5H-furo[3,2-g] [1]-benzopyran-5-one) (iprocrolol);

((-)-5-(tert butylamino)-2-hydroxypropoxy]-3,4-dihydro-1-(2H)-naphthalenone HCl) (levobunolol);

(4-(2-hydroxy-3-isopropylamino-propoxy)-1,2-benzisothiazole HCl);

(4-[3-(tert.butylamino)-2-hydroxypropoxy]-N-methylisocarbostyril HCl) ;

((±)-N-2-[4-(2-hydroxy-3-isopropylaminopropoxy)phenyl]ethyl-N'-isopro pylurea) (pafenolol); (3- [ [ (2-trifluoroacetamido)ethyl] amino]-1-phenoxyproρan-2-ol);

(N- (3-(o-chlorophenoxy)-2-hydroxypropyl)-N'-(4'-chloro-2,3-dihydro-3 oxo-5-pyridazinyl)ethylenediamine);

( (±)-N-[3-acetyl-4-[2-hydroxy-3-[(1-methylethyl)amino]propoxyphenyl]-butanamide) (acebutolol); ((±)-4'-[3-(tert-butylamino)-2-hydroxypropoxy]spiro[cyclohexane-1,2'-indan]-1'-one) (spirendolol);

(7-[ 3-[[2-hydroxy-3-[(2-methylindol-4-yl)oxylpropyl]amino]butyl] thiophylline) (teoprolol); ((±) -1-tert.butylamino-3-(thiochroman-8-yloxy)-2-propanol) (tertatolol);

((±)-1-tert.butylamino-3-(2,3-xylyloxy)-2-propanol HCl) (xibenolol);

(8- [3-(tert. butylamino)-2-hydroxypropoxy]-5-methylcoumarin) (bucumo-1-ol);

(2-(3-(tert.butylamino)-2-hydroxy-propoxy)benzonitrile HCl) (bunitrolol);

( (±) - 2'- [ 3-(tert-butylamino)-2-hydroxypropoxy-5'-fluorobutyrophenone)

(butofilolol); (1-(carbazol-4-yloxy)-3-(isopropylamino)-2-propanol) (carazolol);

(5- (3-tert. butylamino-2-hydroxy) propoxy-3,4-dihydrocarbotyril HCl)

(carteolol);

(1-(tert.butylamino)-3-(2,5-dichlorophenoxy)-2-propanol) (cloranolol); (1-(inden-4(or 7) -yloxy)-3-(isopropylamino)-2-propanol HCl) (indenolol);

(1-isopropylamino-3-[(2-methylindol-4-yl)oxy]-2-propanol) (mepindolol);

(1-(4-acetoxy-2,3,5-trimethylphenoxy)-3-isopropylaminopropan-2-ol) (metipranolol);

(1-(isopropylamino)-3-(o-methoxyphenoxy)-3-[(1-methylethyl) amino]-2-propanol) (moprolol);

((1-tert.butylamino)-3-[(5,6,7,8-tetrahydro-cis-6,7-dihydroxy-1-naphthyl)oxy]-2-propanol) (nadolol); ((S)-1-(2-cyclopentylphenoxy)-3-[(1,1-dimethylethyl) amino]-2-propanol sulfate (2:1)) (penbutolol);

(4'-[1-hydroxy-2-(amino)ethyl]methanesulfonanilIde) (sotalol);

(2-methyl-3-[4-(2-hydroxy-3-tert.butylaminopropoxy) phenyl]-7-aethoxyisoquinolin-1-(2H)-one); (1-(4-(2-(4-fluorophenyloxy)ethoxy)phenoxy)-3-isopropylamino-2-ρroρanol

HCl);

((-)-p-[3-[(3,4-dimethoxyphenethyl)amino]-2-hydroxypropoxy]-β-methylcinnamonltrile) (pacrlnolol);

((±)-2-(3'-tert. butylamino-2'-hydroxypropylthio)-4-(5'-carbamoyl-2'- thienyl)thiazole HCl) (arotinolol);

((±)-1-[p-[2-(cyclopropylmethoxy)ethoxy]phenoxy]-3-(isopropylamino)-2-propancl) (cicloprolol);

((±)-1-[(3-chloro-2-methylindol-4-yl)oxy]-3-[(2-phenoxyethyl)amino]-2- propanol) (indopanolol);

( (±) -6- [ [2- [ [ 3- (p-butoxyphenoxy)-2-hydroxypropyl] amino] ethyl] amino]- 1,3-dimethyluracil) (pirepolol);

(4-(cyclohexylamino)-1-(1-naphtholenyloxy)-2-butanol); (1-phenyl-3- [2- [3- (2 -cyanophenoxy)-2-hydroxypropyl]aminoethyl]hydantoin HCl);

(3 ,4-dihydro-8-(2-hydroxy-3-isopropylaminopropoxy)-3-nitroxy-2H-1-benzopyran) (nipradolol);

Angiotensin I Converting Enzyme Inhibitors: 1-(3-mercapto-2-methyl-1-oxoproρyl)-L-proline (captopril);

(1-(4-ethoxycarbonyl-2,4(R,R)-dimethylbutanoyl) indoline-2(S)-carboxylic acid);

(2-[2-[(1-(ethoxycarbonyl)-3-phenyl-propyl] amino]-1-oxopropyl]-1,2,3,4- tetrahydro-3-isoquinoline carboxylic acid); ((S)-1-[2-[(1-(ethoxycarbonyl)-3-phenylpropyl]amino]-1-oxoρropyl] octahydro-1H-indole-2-carboxylic acid HCl);

(N-cyclopentyl-N-(3-(2,2-dimethyl-1-oxopropyl) thiol-2-methyl-1-oxo¬propyl) glycine) (pivalopril);

( (2R.4R) -2-(2-hydroxyphenyl)-3-(3-mercaptopropionyl)-4-thiazolidinecarboxylic acid);

(1-(N-[1(S)-ethoxycarbonyl-3-phenylpropyl]- (S)-alanyl) -cis, syn-octahydroindol-2(S)-carboxylic acid HCl);

((-)-(S)-1-[(S)-3-mereapto-2-methyl-1-oxopropyl] indoline-2-carboxylic acid); ( [1(S),4S]-1-[3-(benzoylthio)-2-methyl-1-oxoρropyl]-4-phenylthio-L-proline;

(3-([1-ethoxycarbonyl-3-phenyl-(1S)-propyl] amino)-2,3,4,5-tetrahydro-2-oxo-1-(3S)-benzazepine-1-acetic acid HCl);

(N-(2-benzyl-3-mercaptopropanoyl)-S-ethyl-L-cysteine) and the S-methyl analogue;

(N-(1(S)-ethoxycarbonyl-3-phenylpropyl)-L-alanyl-L-proline maleate) (enalapril);

N-[1-(S)-carboxy-3-phenylpropyl]-L-alanyl-1-proline; N -[1-(S)-carboxy-3-phenylpropyl]-L-lysyl-L-proline (lysinopril); Other Antihypertensive Agents: aminophylline; cryptenamine acetates and tannates; deserpidine; meremethoxylline procaine; pargyline; tri-methaphan camsylate; and the like, as well as admixtures and combinations thereof. Typically, the individual daily dosages for these combinations can range from about one-fifth of the minimally recommended clinical dosages to the maximum recommended levels for the entities when they are given singly. Coadministration is most readily accomplished by combining the active ingredients into a suitable unit dosage form containing the proper dosages of each. Other methods of coadministration are, of course, possible.

The compounds of the present invention are prepared as depicted in the charts and as described more fully in the Preparations and Exampies. In the charts, Ph is used to represent the phenyl ring.

Chart A The synthesis of both (Z) and (E) cyclopropylphenylalanine derivatives is reported in the literature and is outlined in Chart A. The (Z) benzyldineoxazoline A-2 is prepared from hippuric acid A-1 via an Org. Syn. procedure, J.S. Buck and W.S. Ide, Org. Syn., Coll. Vol. II, 55 (1943). A standard means of isomerizing (Z) arylidineoxazolones is with saturated hydrobromic acid, Y.S. Rao, Synthesis, 1975:749 (1975). Treatment of the (Z) isomer A-2 suspended in hydrobromic acid saturated with hydrogen bromide at 0ºC in the light overnight affords the (E) isomer A-3.

Various literature reports describe the addition of diazomethane to the benzilidineoxazolones. S.W. King, J.M. Riordan, E.M. Holt, and CH. Stammer, J. Org. Chem., 47:3270 (1982). Also see CH. Stammer, CA 103(25) 215811w. I. Arenal, M. Bernabe, E. Fernandez-Alvarez, M.L. Izquierdo, and S. Penades, J. Heterocyclic Chem., 20:607, 1983; and I. Arenal, M. Bernabe, E. Fernandez-Alvarez, and S. Penades, Synthesis, 1985:773, 1985. Using the procedure of Stammer, excess diazomethane in ether is added to the pure (Z) isomer A-2 in methylene chloride and stirred at ambient temperature overnight. This reaction affords (Z)-2-phenyl-4-(1-methylbenzylidene)-5(4H)-oxazolone A-4, (Z)-1,5-diphenyl-6- oxa-4-azaspiro(2.4) hept-4-en-7-one A-6 as the major product, and, in some cases, (E)-1,5-diphenyl-6-oxa-4-azaspiro(2.4)hept-4-en-7-one A-7. Each of these products is separated by silica gel chromatography using toluene as the eluent. The diazomethane addition to (E) benzilidine oxazolone A-3 affords the (E) cyclopropyl product A-7 as the major product, (E) -2-phenyl-4- (1-methylbenzylidene)-S(4H)-oxazolone A-5, and, in some cases, the (Z) cyclopropyl product A-6. Charts B and C In Chart B, the A-6 compound of Chart A is employed as the B-1 starting material. In Chart C, the A-7 compound of Chart A is employed as the C-1 starting material. For both stereoisomers, the oxazolone B-1 or C-1 is opened with 4- dimethylaminopyridine in methanol to afford the compounds B-2 or C-2. This is followed by removal of the benzoyl group with freshly prepared Meerwein's salt via the procedure of Stammer (S.W. King, J.M. Riordan, E.M. Holt, and CH. Stammer, J. Org. Chem., 47:3270, 1982; also see CH. Stammer, CA, 103(25) : 215811w) to afford the compounds B-3 or C- The amine is protected with a Boc group and the ester saponified to afford the compounds B-4 or C-4.

The cyclopropyl amino acids B-4 or C-4 are incorporated into a peptide using standard coupling procedures. Should the peptide exist in a protected form, the protecting groups are removed prior to coupling. For example, a Boc group is removed from an N-terminus with trifluoroacetic acid in methylene chloride and then the cyclopropyl amino acid is introduced. After coupling, any remaining protecting groups are removed under standard conditions. For example a tosyl group is removed from histidine using 1-hydroxybenzotriazole in dimethylformamide.

Chart D

The phenylalaninol of formula D-1 is commercially available.

Treatment of phenylalaninol of formula D-1 with Boc anhydride in tetrahydrofuran at room temperature affords the compound of formula D-2 as a white crystalline solid. Hydrogenation of the phenyl ring is affected with 5% Rh/Al in ethanol under 50 PSI for two days and affords the compound of formula D-3. Sulfur trioxide (3 equivalents) in DMSO is added to a solution of the alcohol of formula D-3 and triethylamine (TEA) (3 equivalents) in DMSO maintaining the reaction temperature at

25°C to afford the aldehyde of formula D-4. This aldehyde is worked up cold, the solvent evaporated at 30°C, and is quickly used in the next step.

When the crude aldehyde of formula D-4 is added to the Grignard reagent [of 1-bromo-2-methylproρene at -10°C, the allylic alcohol of formula D-5 is obtained after column chromatography. This material by¹ H-NMR and ¹³C-NMR is a 6/1 threo/erythio mixture of diastereomers.

The allylic alcohol of formula D-5 is protected as its acetonide of formula D-6 by standard procedures.

Cyclopropanation by adding 5 equivalents of ethyl diazoacetate to the neat olefin of formula D-6 and anhydrous copper or palladium catalyst at 90ºC affords five diastereomeric cyclopropane products. These cyclopropanes are separated into 3 fractions via chromatography, which are referred to as fractions A, B and C from least to most polar. Fraction A is a single diastereomer. This material is a waxy solid with a melting point of 50-64°C Fraction B is a 5/2 mixture of two diastereomers designated b₁ and b₂. Fraction G which is also a solid is a 7/5 mixture of two diastereomers designated c₁ and c₂. All of these fractions have M+1 molecular ions in their FAB mass spectra with the theoretical high resolution mass. Their IR and ¹H-NMR look practically identical. The separate diastereomers can be observed in the ¹³C-NMR. The two major diastereomeric cyclopropane products are formula D-7a and D-7b1, with the more polar one being preferred for the further reactions described below.

Any of these diastereomeric cyclopropane products can be utilized in the following reaction steps to achieve the peptides of this invention. The trans diastereomeric cyclopropane products are preferred.

Fraction B is treated with catalytic tosic acid in methanol at 60°C to remove the acetonide and to afford the alcohol of formula D-8. The compound of formula D-8 is saponified using 50% methanol/2N LiOH (aq) at 40°C to afford the acid of formula D-9. The D-9 acid and isoleucyl-2-pyridyl-methylamide are coupled in dimethylformamide (DMF) using diethylcyanophosphonate and triethylamine (TEA) to afford the D-10 compound. The D-10 compound is dissolved in methylene chloride/trifluoroacetic acid (TFA) at -20°C and stirred for 19 h. Work-up with saturated sodium bicarbonate yields the amino intermediate of formula D-11 which is coupled to Boc-His(Tos) in DMF in the presence of diethylcyanophosphonate and TEA to yield the compound of formula D-12.

Deprotection of the compound of formula D-12 is conducted in the same manner as described above for the compound of formula D-10 and the resulting amine of formula D-13 is coupled to Boc-Phe, in the same manner as described above for the compound of formula D-11, to yield the compound of formula D-14.

The Tos group is removed from the compound of formula D-14 by treatment with 1-hydroxybenzotriazole in methanol to yield the finnal product of formula D-15.

Chart E Chart E describes the preparation of peptides of this invention containing two cyclopropyl amino acid groups.

The intermediate of formula E-l, prepared in Chart D as the compound of formula D-13, is coupled to (Z) -2-phenyl-1-(Boc)aminocyclopropane-1-carboxylic acid in the presence of diethylcyanophosphonate and diisopropylethylamine in methylene chloride to yield two major isomers of the compound of formula E-2. (Both R and S are present). The more polar isomer is treated with 1-hydroxybenzotriazole in methanol to effect removal of the Tos group to yield the compound of formula E-3.

Chart F The synthesis of the cyclobutane transition state inserts originates with the olefin of formula F-1, prepared as the compound of formula D-5 in Chart D. Conversion of the compound of formula F-1 to the allylic ester of formula F-2 is accomplished in straightforward fashion via esterification. The compound of formula F-2 is submitted to a photochemical 2+2 cycloaddition. M. Tanaka et al., Tetrahedron Letters (1985) 3035. Both the compounds of formula F-3a and F-3b are formed in the cycloadditon reaction and are separable by chromatographic means. Treatment of the compound of formula F-3a with hydroxide first opens the lactone and secondly results in the epimerization of the acid group to give the desired product of formula F-4a (syn isomer). Treatment of the crude reaction with anhydrous acid then effects lactonization of any cis hydroxy acid that may be present and thus provides a means of separating isomers. The acid of formula F-4a is then ready for further C- terminal coupling. Treatment of the compound of formula F-3b under identical conditions leads to the compound of formula F-4b (anti isomer). Following procedures analogous to those described in Chart D starting withi, the compound of formula D-9, the compounds of formula F-4a and F-4b can be converted to the peptides of the present invention. Chart G

Chart G illustrates an additional method for the synthesis of the cyclobutane transition state inserts. The trans olefin of formula G-2 is generated via a Wittig reaction, i.e., reaction of the compound of formula G-1, prepared as the compound of formula D-4 in Chart D, with Ph₃P=C(CH₂)₃CO₂R. Epoxidation of the compound of formula G-2, using the directing influence of the protected nitrogen, provides the desired epoxide of formula G-3. Deprotonation of the compound of formula G-3 effects an intramolecular cyclization giving rise to the desired target "compound of formula G-4. Following procedures analogous to those described in Chart D starting with the compound of formula D-9, the compound of formula G-4 (wherein R is hydrogen) can be converted to the peptides of the present invention. Also, following procedures analogous to those described in Charts G and D, peptides of the present invention containing cyclopentyl and cyclohexyl transition state inserts may be prepared.

Chart H Chart H illustrates another method for the synthesis of the cyclobutane transition state inserts. Addition of ylid P(Ph)₃=CCH₂C≡C to the compound of formula H-1, prepared as the compound of formula D-4 In Chart D, affords the enyne of formula H-2. Carbomethoxylation o the compound of formula H-2 gives the compound of formula H-3 whic upon addition of lithium dimethyl cuprate, and then a higher order cuprate, B.H. Lipshutz et al., Tetrahedron (1984) 40, 5005 and references therein, for the introduction of the second methyl group, yields the compound of formula H-4. Direct epoxidation of the olefin of formula H-4, or epoxide introduction via iodocarbamatlon, M. Hirama et al., Tetrahedron Letters (1984) 4963, to give an intermediate such as the compound of formula H-5, provides the desired epoxide of formula H- 6 with the correct stereochemistry. Cyclization of the compound of formula H-6 then affords the final product of formula H-7. Following procedures analogous to those described In Chart D starting with the compound of formula D-9, the compound of formula H-7 (wherein R is hydrogen) can be converted to the peptides of the present invention.

Charts G and H described above provide entry to the syn series. To approach the anti series, the cis olefins corresponding to formula G-2 and H-2 must be used.

In the cyclopropane, cyclobutane, cyclopentane, and cyclohexane cases, these transition state inserts can readily be coupled to amino acids to build the desired peptides through the use of standard procedures.

Generally, the renin Inhibiting polypeptides may be prepared by either polymer assisted or solution phase peptide synthetic procedures analogous to those described hereinafter or to those methods known in the art. For example, the carboxylic moiety of N^α- t-butyloxycarbonyl (Boc)-substituted amino acid derivatives having suitable side chain protecting groups, if necessary, may be condensed with the amino functionality of a suitably protected amino acid, peptide or polymer-bound peptide using a conventional coupling protocol such as dicyclohexylcarbodiimide (DCC) and 1-hydroxybenzotriazole (HOBT) or diethylphosphoryl cyanide (DEPC) and triethylamine (Et₃N) in methylene chloride or dimethylformamide. The synthetic procedures used to incorporate the novel moieties herein are analgous to those described, for example, in U.S. patents 4,424,207; 4,470,971; 4,477,440; 4,477,441; 4,478,826; 4,478,827; 4,479,941; and 4,485,099, and copending application Serial No. 753,198, filed 9 July 1985, and copending application Serial No. 825,250, filed 3 February 1986, all of which are expressly incorporated by reference herein. See, also, published European patent applications 45,161; 45,665; 53,017; 77,028; 77,029; 81,783; 104,041; 111,266; 114,993; and 118,223.

Following coupling reaction completion, the N^α-Boc moiety may be selectively removed with 45% trifluoroacetlc acid with or without 2% anisole (v/v) in methylene chloride. Neutralization of the resultant trifluoroacetate salt may be accomplished with 10% diisopropylethylamine or sodium bicarbonate in methylene chloride. In the case of polymer-assisted peptide synthesis, this stepwise, coupling strategy may be partially or completely automated to provide the desired peptide-polymer intermediates. Anhydrous hydrofluoric acid treatment of the peptide-polymer intermediate may then be used to effect simultaneous protecting group removal and cleavage of the peptide from its polymeric support. A notable exception to this includes Nⁱⁿ-formyl- indolyl-substituted peptides in which the Nⁱⁿ-formyl- indolyl moiety is stable to TFA or HF but may be removed by NH₃ or NaOH. Because FTrp is somewhat unstable to base in synthetic procedures, possibly causing lower yields, it may be desirable in solution phase synthesis to introduce the FTrp-containing moiety late in the synthetic sequence so that it is not exposed to such conditions.

The incorporation of Nⁱⁿ-formyl-Trp into compounds of the present invention is easily accomplished because of the commercial availability of N^α-Boc-Nⁱⁿ-formyl-Trp-OH. However, the Nⁱⁿ-formyl moiety may be introduced into indolyl-substituted amino acid derivatives or related compounds by reaction with HCl-formic acid as reported in the literature, see A. Previero et al, Biochim. Biophys. Acta 147, 453 (1967); Y.C ang et al, Int. J. Peptide Protein Res. 15, 130 (1980). Generally, methods of alkylation useful in alkylating histidine for use in the present invention are found in Cheung, S.T. et al, Can. J. Chem., Vol 55, pp. 906-910 (1977). However it is now found that in Cheung, S. T. et al, methods it is critical that the reaction conditions for the alkylation of histidine be anhydrous. Further, it is now found also that during work-up instead of adding water directly to the reaction mixture, It is preferred that a buffered aqueous solution be added to the reaction mixture, for example, aqueous sodium or potassium hydrogen sulfate.

Variations in the above description for starting materials, reactants, reaction conditions and required protecting groups to obtain other such N-alkylated compounds are known to an ordinarily skilled chemist or are readily available in the literature.

These peptides may also be prepared by the standard solid phase techniques of Merrifleld. Appropriate protecting groups, reagents, and solvents for both the solution and solid phase methods can be found in "The Peptides: Analysis, Synthesis, and Biology," Vols. 1-5, eds. E. Gross and T. Meienhofer, Academic Press, NY, 1979-1983.

The compounds of the present invention may be in either free form or In protected form at one or more of the remaining (not previously protected) peptide, carboxyl, amino, hydroxy, or other reactive groups. The protecting groups may be any of those known in the polypeptide art. Examples of nitrogen and oxygen protection groups are set forth in T.W. Greene, Protecting Groups in Organic Synthesis, Wiley, New York, (1981); J.F.W. McOmie, ed. Protective Groups in Organic Chemistry, Plenum Press (1973); and J. Fuhrhop and G. Benzlin, Organic Synthesis, Verlag Chemie (1983). Included among the nitrogen protective groups are t-butoxycarbonyl (Boc), benzyloxycarbonyl, acetyl, allyl, phthalyl, benzyl, benzoyl, trityl and the like. DESCRIPTION OF THE PREFERRED EMBODIMENTS The following Preparations and Examples illustrate the present invention.

In the Preparations and Examples below and throughout this document: Ac is acetyl;

AMP is 2-(aminomethyl)pyridinyl; BOC is t-butoxycarbonyl; BOM is benzyloxymethyl; Bz is benzyl;

C is centigrade; Celite is a filter aid;

CVDA is "Cha-Val diol" where Cha is 3-cyclohexylalanyl, i.e. the moiety of the formula XL₆ wherein R₁ is cyclohexyl and R₁₁ is isopropyl and the configuration at each carbon atom with a * is (R);

DCC is dicyclohexylcarbodiimide;

DMF is dimethylformamide;

EtOAc is ethyl acetate; g. is grams;

GEA is 2- (quanidylethyl)amino;

GMPMA is (3-(guanidylmethyl)phenyl)methylamino;

HPLC is high performance liquid chromatography;

I₂ is iodine; IR is infra red spectra;

A Lindlar catalyst is a modified 5% palladium on calcium carbonate catalyst, obtained from Engelhard Industries and used for reduction;

LVDA is "Leu-Val diol," i.e., the moiety of the formula XL₆ wherein R₁ and R₁₁ are isopropyl and the configuration at each carbon atom with a * is (R) ;

M or mol is mole;

MBA is 2-methylbutylamino (racemic or optically active);

MBAS is 2S-methylbutylamino; Me is methyl; min. is minute; ml is milliliter;

MS is mass spectroscopy;

NMHis is Nα-methyl-L-histidine; NMR is nuclear magnetic resonance;

NOAl is (1-naphthyloxy)acetyl; p-TSA salt is para-toluene sulfonic acid salt;

Ph is phenyl; POA is phenoxyacetyl;

RIP means a compound having the formula H-Pro-His-Phe-His-Phe- Phe-Val-Tyr-Lys-OH.2(CH₃C(O)OH).XH₂O which is a known renin- inhibiting peptide. Skellysolve B is as defined in the Merck Index, 10th edition;

TBDMS is t-butyldimethylsilyl; TFA is trifluoroacetic acid; THF is tetrahydrofuran; TLC is thin layer chromatography; Tos is p-toluenesulfonyl;

Tr is trityl (triphenylmethyl); 2HPA is (±)-(2-hydroxypropyl)amino; and UV is ultraviolet.

The wedge-shape line indicates a bond which extends above the plane of the paper relative to the plane of the compound thereon. The dotted line indicates a bond which extends below the plane of the paper relative to the plane of the compound thereon. Preparation 1 (Z) 2-Phenyl-4-benzylidene-5(4H)-oxazolone (Formula A- 2) Refer to Chart A.

Benzaldehyde (51 g), hlppuric acid (96 g), and sodium acetate (arih. 40 g) are suspended in acetic anhydride (140 ml) in a 1 1 Ehrlenmeyer flask. This is heated on a hot plate with shaking. After becoming a solid mass, it begins liquifying. Once all of the material is a yellow liquid, the flask is maintained at 100° C for 2 hours. Ethanol (200 ml) is added at 0°C. After sitting overnight, the crystals are collected and washed two times with 50 ml of cold ethanol and boiling water to afford 88.25 g of crude material.

This yellow solid is dissolved in hot benzene (600 ml) and filtered through sodium sulfate. After sitting at 4ºC, 64.5 g of yellow crystalline title product is collected. Physical characteristics are as follows: Mp: 163-166°C

Silica gel TLC: R_f=0.5 in benzene. IR (cm^-1, mull): 2954, 2926, 2855, 1797, 1655, 1492, 1450, 1329, 1297, 1169, 869, 768, 699, 687. 1H-NMR (δ , CDCI₃): 8.25-8.11, 7.53-7.41, 7.23.

UV (λ_maχ): 223, 240, 246, 261, 292, 330, 348, 362, 381. Mass spectrum found: 249.0793; (ions at m/e) : 250, 249, 116, 106, 105, 89, 77, 51, 38.

Anal, found: C, 76.58; H, 4.55; N, 5.54. Preparation 2 (E) 2-Phenyl-4-benzylidene-5(4H)-oxazolone (Formula A - 3) Refer to Chart A .

Hydrobromic acid (167 ml) is added to the (Z) -oxazolone of Preparation 1 (10 g) at 0°C Anhydrous hydrogen bromide is bubbled through this suspension for 30 min. After standing at 4°C under light overnight, the reaction suspension is poured over ice, filtered and the yellow solid washed with ice water. The solid is dissolved in ethyl acetate and dried over magnesium sulfate to afford 9.2 g of the title product.

Physical characteristics are as follows: Mp: 145-147°C Silica gel TLC: R_f=0.4 in benzene.

IR (cm^-1, mull): 2954, 2926, 2870, 2855, 1799, 1772, 1647, 1452, 1369, 1326, 1296, 1227, 1156, 1118, 1008, 899, 777, 760, 700, 685, 636. 1H-NMR (δ, CDCl₃): 8.21-8.02, 7.56-7.25. UV (λ_max): 225, 239, 246, 261, 292, 348, 362, 380. Mass spectrum (ions at m/e): 249, 116, 106, 105, 89, 77, 76, 63, 51, 50.

Anal found: C, 76.25; H, 4.56; N, 5.53. Preparation 3 (Z) -2-Phenyl-4-(1-methylbenzylidene)-5(4H)-oxazolone, (Z) -1,5-Diphenyl-6-oxa-4-azaspiro(2.4)hept-4-en-7-one, and (E)-1,5-Diphenyl-6-oxa-4-azaspiro(2.4)hept-4-en-7- one (Formulas A-4, A-6 and A-7) Refer to Chart A. Diazomethane (formed from 30 g of N-methyl-N'-nitro-N-nitrosoguanidine over 160 ml 45% potassium hydroxide in 400 ml ether and dried over potassium hydroxide pellets) is added to a solution of the title product of Preparation 1 (12 g) in methylene chloride (200 ml) at ambient temperature. Nitrogen evolution is immediate. After stirring overnight, excess diazomethane is destroyed by adding solid calcium chloride. Filtration and evaporation of the filtrate affords 19.8 g of crude product. Flash chromatography over silica gel (1.2 kg, toluene) affords 1.24 g of a mixture of starting material and the first title product. Further elution affords 2.59 g of the first title product. An analytical sample is prepared by recrystallization from methylene chloride to give yellow crystals.

Physical characteristics are as follows:

Mp: 101-104°C

Silica gel TLC: R_f=0.45 in benzene. IR (cm^-1, mull): 2954, 2926, 2869, 2856, 1784, 1761, 1649, 1637, 1452, 1329, 1299, 1194, 971, 768, 701.

¹H-NMR (δ, CDCl₃): 8.12-8.09, 8.05-8.00, 7.52-7.19, 2.79.

UV (λ_maχ): 242sh, 247, 255, 334sh, 345.

Mass spectrum found: 263.0953; (ions at m/e): 263, 106, 105, 103, 78, 77, 76, 63, 51, 50.

Anal, found: C, 77.21; H, 4.89; N, 5.26.

Further elution affords 5.36 g of the second title product. An analytical sample is prepared by recrystallization from methylene chloride/hexane to give the crystalline form. Physical characteristics are as follows:

Mp: 145-146°C

Silica gel TLC: R_f=0.4 in benzene.

IR (cm^-1, mull).: 2954, 2925, 2855, 1805, 1635, 1453, 1322, 1255, 1017, 994, 985, 877, 710, 696, 689. 1H-NMR (δ, CDCI₃): 7.98-7.85, 7.47-7.30, 3.22, 2.36, 2.25.

UV (λ_max): 205, 236, 264, 270.

Mass spectrum (ions at m/e): 263, 219, 218, 130, 115, 106, 105, 78, 77, 51.

Anal, found: C, 77.14; H, 4.99; N, 5.32. Further elution affords 1.90 g of the third title product. An analytical sample is prepared by recrystallization from methylene chloride/hexane to give the crystalline form.

Physical characteristics are as follows:

Mp: 119-120° C Silica gel TLC: R_f=0.25 in benzene.

IR (cm^-1, mull): 2954, 2925, 2855, 1805, 1797, 1636, 1451, 1323, 1303, 1036, 1016, 1001, 981, 874, 776, 720, 692. 1H-NMR (δ, CDCI₃): 8.04-7.92, 7.53-7.45, 7.32, 3.53, 2.44, 2.32.

UV (λ_max): 270. Mass spectrum (ions at m/e): 263, 219, 130, 115, 106, 105, 103, 78, 77, 51.

Anal, found: C, 77.21; H, 5.13; N, 5.24. Preparation 4 (E) -2-Phenyl-4-(1-methyϊbenzylidene)-5(4H)-oxazolone, and (E) -1,5-Diphenyl-6-oxa-4-azaspiro(2.4)hept-4-en-7- one (Formulas A- 5 and A-7) Refer to Chart A.

Diazomethane (formed from 30 g of N-methyl-N'-nitro-N-nitrosoguanidine over 160 ml 45% potassium hydroxide in 400 ml ether and dried over potassium hydroxide pellets) is added to a solution of the title product of Preparation 2 (12 g) in methylene chloride (200 ml) at ambient temperature. Nitrogen evolution is immediate. After stirring overnight, excess diazomethane is destroyed by adding solid calcium chloride. Filtration and evaporation of the filtrate affords 15.1 g of crude product.

Flash chromatography over silica gel (800 g, toluene) affords 0.58 g of the first title product. An analytical sample is prepared by recrystallization from methylene chloride/hexane to give the crystalline form. Physical characteristics are as follows: Mp: 114-115°C

Silica gel TLC: R_f=0.4 in benzene.

IR (cm^-1, mull): 2954, 2925, 2869, 2855, 1790, 1653, 1450, 1442, 1290, 1202, 1059, 883, 763, 695, 685. 1H-NMR (δ, CDCl₃): 8.16-8.04, 7.59-7.35, 2.66. UV (λ_max): 239, 245, 341.

Mass spectrum (ions at m/e): 264, 263, 106, 105, 103, 78, 77, 76, 51, 50.

Anal, found: C, 77.17; H, 5.17; N, 5.35. Further elution affords 8.05 g of the second title product.

Preparation 5 Methyl (Z) -2 -phenyl-1-benzamidocyclopropane-1-carboxylate (Formula B-2) Refer to Chart B.

4-Dimethylaminopyridine (5.32 g) is added to a solution of the second title product of Preparation 3 (11.2 g) in absolute methanol (300 ml). After stirring at ambient temperature until everything is in solution (110 min), the solution is evaporated in vacuo. The residue is transferred in methylene chloride to a separatαry funnel containing

5% citric acid. Extracting two times with methylene chloride and filtering the combined organic layers through sodium sulfate affords 13.0 g of crude product after evaporation of the solvent.

Recrystallization from methylene chloride/hexane affords 10.68 g of the title product. Flash column chromatography of the mother liquors (200 g silica gel, ether) affords an additional 0.73 g. Physical characteristics are as follows: Mp: 164-166.5°C Silica gel TLC: R_f=0.5 in ether.

IR (cm^-1, mull): 3275, 2950, 2923, 2867, 2855, 1716, 1667, 1648, 1552, 1455, 1437, 1338, 1274, 1162, 713, 696.

¹H-NMR (δ, CDCl₃): 7.40-7.25, 5.^'96, 3.76, 3.05, 2.33, 1.86. UV (λ_maχ): 224, 271sl sh.

Mass spectrum (ions at m/e): 295, 263, 191, 190, 130, 106, 105, 104, 103, 78, 77, 51. Anal, found: C, 72.74; H, 5.76; N, 4.72.

Preparation 6 Methyl (E) -2-phenyl-1-benzamidocyclopropane-1-carboxy late (Formula C-2) Refer to Chart C.

4-DimethylamInoρyridine (6.33 g) is added to a solution of the second title product of Preparation 4 (13.3 g) in absolute methanol (360 ml). After stirring at ambient temperature until everything is in solution (130 min), the solution is evaporated in vacuo. The residue is transferred in methylene chloride to a separatory funnel containing

5% citric acid. Extracting two times with methylene chloride and filtering the combined organic layers through sodium sulfate affords 14.8 g of crude product after evaporation of the solvent.

Recrystallization from methylene chloride/hexane affords 12.44 g of the title product. Flash column chromatography of the mother liquors (200 g silica gel, ether) followed by recrystallization of the fractions containing the title product affords an additional 0.26 g. Physical characteristics are as follows: Mp: 194-197°C

Silica gel TLC: R_f=0.45 in ether.

IR (cm^-1, mull): 3326, 2957, 2924, 2870, 2855, 1734, 1636, 1580, 1529, 1489, 1438, 1350, 1220, 1196, 1163, 1154, 730, 718, 701. 1H-NMR (δ, CDCI₃): 7.90-7.77, 7.50-7.21, 6.94, 3.35, 2.98, 2.32, 1.73.

UV (λ_maχ): 224.

Mass spectrum (ions at m/e): 295, 191, 190, 130, 106, 105, 104, 103, 78, 77, 51. Anal, found: C, 72.80; H, 5.92; N, 4.63.

Preparation 7 Methyl (Z)-2-phenyl-1-aminocyclopropane-1-carboxylate, hydrochloride (Formula B-3) Refer to Chart B. A solution of triethyloxonium tetrafluoroborate (Meerwein's salt, freshly prepared and stored under ether at -15°C, 8.86 g) in methylene chloride (26 ml) is placed in a flame dried flask under nitrogen. To this solution is added a suspension of the title product of Preparation 5 (3.79 g) in methylene chloride (36 ml). After refluxing for 19 h under nitrogen, the reaction solution is transferred to a separatory funnel containing 0,93 M potassium dihydrogen phosphate (27 ml). Extracting with methylene chloride and drying over magnesium sulfate affords 4.42 g of crude amine product after evaporation of the solvent. This is dissolved in ether (27 ml) and cooled to -25°C before adding hvdrochloride saturated ether (20 ml) and 1 N hydrochloric acid (55 ml). After stirring at room temperature for 10 min, the aqueous layer is extracted twice with ether and lypholized to afford 2.79 g of crude product. Recrystallization from acetonitrile/ether affords 2.00 g of the title product. Physical characteristics are as follows: Mp: 186-188°C

IR (cm^-1, mull): 2954, 2920, 2869, 2855, 2820, 2680, 1734, 1567, 1461, 1440, 1312, 1211, 770, 722. 1H-NMR (δ, CDCl₃): 7.59, 3.84, 3.11, 2.19, 2.00. UV (λ_maχ): 218, 252, 259, 264.

Mass spectrum (ions at m/e): 191, 159, 132, 131, 130, 117, 115, 104, 103, 91, 77.

Anal, found: C, 57.92; H, 6.06; N, 6.21. Preparation 8 Methyl (E) -2-phenyl-1-aminocyclopropane-1-carboxylate, hydrochloride (Formula C-3) Refer to Chart C.

A solution of triethyloxonium tetrafluoroborate (Meerwein's salt, freshly prepared and stored under ether at -15ºC, 8.6 g) in methylene chloride (25 ml) is placed in a flame dried flask under nitrogen. To this solution is added a suspension of the title product of Preparation 6 (3.68 g) in methylene chloride (35 al) . After refluxing for 23 h under nitrogen, the reaction solution is transferred to a separatory funnel containing 0.93 M potassium dihydrogen phosphate (27 ml). Extracting with methylene chloride and drying over magnesium sulfate affords 4.59 g of crude amine product after evaporation of the solvent. This is dissolved in ether (27 ml) and cooled to -25°C before adding hydrochloride saturated ether (20 ml) and 1 N hydrochloric acid (54 ml). After stirring at room temperature for 1 h, the aqueous layer is extracted twice with ether and lypholized to afford 2.60 g of crude product. Recrystallization from isopropanol/ether affords 2.35 g of the title product.

Physical characteristics are as follows: Mp: 185-188ºC. IR (cm^-1, mull): 2953, 2924, 2867, 2855, 1754, 1458, 1447, 1370, 1248, 1200, 1170, 773. 1H-NMR (δ, CDCl₃): 7.27, 3.39, 3.42, 2.27, 2.15, 1.72. UV (λ_maχ): 247, 254, 260, 266.

Mass spectrum (ions at m/e): 191, 159, 132, 131, 130, 115, 104, 103, 91, 77, 51.

Anal, found: C, 57.81; H, 6.32; N, 6.25. Preparation 9 (Z) -2-Phenyl-1-(tert-butyloxycarbonyl)aminocyclopropane- 1-carboxylic acid (Formula B-4) Refer to Chart B. A solution of the title product of Preparation 7 (3.47 g) in dioxane (106 ml) and water (45 ml) is adjusted to pH 8.0 with 2N sodium hydroxide. Di-t-butyl dicarbonate (6.62 g) is added portionwise over 1.5 h maintaining the pH at 8.0-8.5 with 2N sodium hydroxide as necessary. After stirring at ambient temperature for 22 h, the solution is cooled to 0°C and extracted two times with ether. Washing the combined organic layers with brine and filtering through sodium sulfate affords 6.46 g of crude boc methyl ester after evaporation of the solvent.

Physical characteristics are as follows:

¹H-NMR (δ, CDCI₃): 7.26, 4.56, 3.75, 2.94, 2.04, 1.65, 1.33. The crude methyl ester is dissolved in methanol (38 ml) and 2N sodium hydroxide (38 ml). After stirring at ambient temperature for 21 h, the reaction solution is reduced to half volume in vacuo and the residue transferred to a separatory funnel with ethyl acetate/water. Acidification with 2N hydrochloric acid to pH 5, extracting with ethyl acetate, washing the combined organic layers with brine, and filtering through sodium sulfate affords 3.14 g of the title product after evaporation of the solvent.

Physical characteristics are as follows: Mp: 200-203ºC IR (cm^-1, mull): 3345, 2954, 2926, 2870, 2855, 1691, 1657, 1649, 1460, 1453, 1444, 1410, 1367, 1270, 1170, 704. 1H-NMR (δ, CDCI₃): 7.25, 4.68, 3.03, 2.13, 1.73, 1.32. UV (λ_maχ): 224, 259, 266, 273. Mass spectrum (ions at m/e): 221, 203, 159, 132, 131, 130, 104, 103, 57, 41.

Anal, found: C, 64.88; H, 6.88; N, 4.85. Preparation 10 (E) -2-Phenyl-1-(tert-butyloxycarbonyl)aminocyclopropane- 1-carboxylic acid (Formula C-4) Refer to Chart C.

A solution of the title product of Preparation 8 (0.60 g) in dioxane (18 ml) and water (7.8 ml) is adjusted to pH 8.0 with 2N sodium hydroxide. Di-t-butyl dicarbonate (1.14 g) is added portionwise over 1 h maintaining the pH at 8.0-8.5 with 2N sodium hydroxide as necessary. After stirring at ambient temperature for 21 h, the solution is cooled to 0°C and extracted two times with ether. Washing the combined organic layers with brine and filtering through sodium sulfate affords 1.01 g of crude boc methyl ester after evaporation of the solvent. Physical characteristics are as follows: 1H-NMR (δ, CDCl₃): 7.26, 5.29, 3.69, 2.83, 2.17, 1.61, 1.48.

The crude methyl ester is dissolved in methanol (6.5 ml) and 2N sodium hydroxide (6.5 ml). After stirring at ambient temperature for 23 h, the reaction solution is reduced to half volume in vacuo and the residue transferred to a separatory funnel with ethyl acetate/water. Acidification with 2N hydrochloric acid to pH 5, extracting with ethyl acetate, washing the combined organic layers with brine, and filtering through sodium sulfate affords 0.60 g of the title product after evaporation of the solvent. An analytical sample is prepared by recrystallization from ethyl acetate/hexane. Physical characteristics are as follows: Mp : 163-173°C decomposed.

IR (cm^-1, mull): 3373, 2954, 2925, 2869, 2855, 1694, 1512, 1457, 1453, 1310, 1252, 1164, 700. 1H-NMR (δ, CDCI₃): 7.25, 3.30, 2.78, 2.15, 1.65, 1.48. UV (λ_maχ): 221, 258sh, 266, 273.

Mass spectrum (ions at m/e): 221, 203, 159, 132, 131, 130, 77, 59, 57, 41.

Anal, found: C, 64.96; H, 6.96; N, 5.30. Example 1 N^α- [ (2S ,4S ,5S) -5- [N- [N^α- [ (E) -2-Phenyl-1-(tert-butoxycarbonylamino) cyclopropane-1-carbonyl]-L-histidyl]- amino]-4-hydroxy-2-isopropyl-7-methyl-1-oxooctyl]-N-(2- pyridinylmethyl)-L-isoleucinamide (Isomers A and B) 1. A solution of N-tert-butyloxycarbonyl-N^im-tosyl-L-histidyl- 5S-amino-2S-isopropyl-7-methyl-4S-tert-butyldiaethylsilyloxyoctanoyl-L-isoleucyl-2-pyridylmethylamide (see EP 0173.481A2, p. 100, C81) (0.068 g) in methylene chloride (0.5 ml), under a nitrogen atmosphere, is cooled in an ice bath and treated dropwise with trifluoroacetic acid (0.5 ml). The mixture is allowed to warm to room temperature and stirred for 1 hr, 5 mln; it is then concentrated in vacuo. The residue is dissolved in methylene chloride, washed with aqueous sodium bicarbonate, dried over magnesium sulfate and concentrated to give 0.047 g of a gummy residue, N^im-tosyl-L-histidyl-5S-amino-4S-hydroxy-2S-isoproρyl-7-methyloctanoyl-L-isoleucyl-2-pyrIdylmethylamide, which is pure by TLC on silica gel with 5% methanol-0.5% ammonium hydroxidemethylene chloride.

2. A solution of the product prepared in paragraph 1 (0.047 g) in methylene chloride (6.5 ml) under a nitrogen atmosphere is treated with 0.025 g of d,1-(E)-2-phenyl-1-(tert-butoxycarbonyl)aminocyclopropane-1-carboxylic acid (prepared in Preparation 10) and then successively with triethylamlne (0.013 ml) and diethylcyanophosphonate (0.015 ml). The mixture is stirred at room temperature for 3 hr, 45 min, diluted with methylene chloride, washed with aqueous sodium bicarbonate, dried with magnesium sulfate and concentrated to give 0.075 g of a gummy residue. This material, is chromatographed on silica gel (50 g) with 3.5% methanol-0.35% ammonium hydroxide-methylene chloride; 4.7 ml fractions are collected. isomer A is eluted from the column first in fractions 100-144 and amounts to 0.018 g. The high resolution FAB mass spectrum has [M + H]⁺ at m/z 985. Theory for C₅₂H₇₃N₈O₉S: 985.5221. Measured: 985.5243.

Isomer B is eluted from the column in fractions 145-250 and amounts to 0.020 g. The high resolution FAB mass spectrum has [M + H]⁺ at m/z 985. Theory for C₅₂H₇₃N₈O₉S: 985.5221. Measured: 985.5195. 3. A solution of the product (Isomer A) prepared In paragraph 2 (0.018 g) in dimethylformamide (0.21 ml) and tetrahydrofuran (1.2 ml) under a nitrogen atmosphere is treated with 1-hydroxybenzotriazole (0.025 g) and stirred at room temperature for 20 hr. The mixture is concentrated in vacuo and the residue is chromatographed on silica gel (25 g) with 5% methanol-0.5% ammonium hydroxide-raethylene chloride; 5.4 ml fractions are collected. The titled product (Isomer A) is eluted in fractions 78-118 and amounts to 0.014 g. The high resolution FAB mass spectrum has [M + H]⁺ at m/z 831. Theory for C₄₅H₆₇N₈O₇: 831.5132. Measured: 831.5112.

4. A solution of the product (Isomer B) prepared in paragrap 2 (0.020 g) in dimethylformamide (0.24 ml) and tetrahydrofuran (1.4 ml) under a nitrogen atmosphere is treated with 1-hydroxybenzotriazole (0.028 g) and stirred at room temperature for 21.5 hr. The mixture is concentrated in vacuo to give a gummy residue (0.051 g) which is chromatographed on silica gel (25 g) with 5% methanol-0.5% ammonium hydroxide-methylene chloride; 5.4 ml fractions are collected. titled product (Isomer B) is eluted in fractions 104-146 and amounts to 0.010 g. The high resolution FAB mass spectrum has [M + H]⁺ at m/z 831. Theory for C₄₅H₆₇N₈O₇: 831.5132. Measured: 831.5112. Example 2 N^α- [ (2S , 4S , 5S) -5- [N- [N^α- [ (Z) -2 -Phenyl-1- (tert-butoxycarbonylamino) cyclopropane-1-carbonyl]-L-histidyl]- amino]-4-hydroxy-2-isopropyl-7-methyl-1-oxooctyl]-N-(2- pyridinylmethyl)-L-isoleucinamide (Isomers A and B)

1. A solution of N-tert-butyloxycarbonyl-N^im-tosyl-L-histidyl- 5S - amino -2S- isopropyl-7-methyl-4S-tert-butyldimethylsilyloxyoctanoyl- L-isoleucyl-2-pyridylmethylamide (see EP 0173.481A2, p. 100, C81) (0.068 g) in methylene chloride (0.5 ml), under a nitrogen atmosphere, is cooled in an ice bath and treated dropwise with trifluoroacetic acid (0.5 ml). The mixture is allowed to warm to room temperature and stirred for 1 hr, 5 min; it is then concentrated in vacuo. The residue is dissolved in methylene chloride, washed with aqueous sodium bicarbonate, dried over magnesium sulfate and concentrated to give 0.039 g of a gummy residue, N^im-tosyl-L-histidyl-5S-amino-4S-hydroxy-2S-isopropyl-7-methyloctanoyl-L-isoleucyl-2-pyridylmethylamide, which is pure by TLC on silica gel with 5% methanol-0.5% ammonium hydroxidemethylene chloride.

2. A solution of the product prepared in paragraph 1 (0.039 g) in methylene chloride (6.5 ml) under a nitrogen atmosphere is treated with

0.025 g of d, 1-(Z)-2-phenyl-1-(tert-butyloxycarbonyl)aminocyclopropane- 1-carboxylic acid (prepared in Preparation 9) and then successively with triethylamine (0.013 ml) and diethylcyanophosphonate (0.015 ml). The mixture is stirred at room temperature for 2 hr, 20 min, diluted with methylene chloride, washed with aqueous sodium bicarbonate, dried over magnesium sulfate and concentrated to give 0.066 g of a gummy residue. This material is chromatographed on silica gel (50 g) with 3.5% methanol-0.35% ammonium hydroxide-methylene chloride; 5 ml fractions are collected.

Isomer A is eluted in fractions 106-184 and amounts to 0.011 g. Isomer B is eluted in fractions 198-330 and also amounts to 0.011 g. 3. A solution of the product (Isomer A) prepared in paragraph 2

(0.011 g) in dimethylformamide (0.13 ml) and tetrahydrofuran (0.72 ml) under a nitrogen atmosphere is treated with 1-hydroxybenzotriazole

(0.015 g) and stirred at room temperature for 17.5 hr. The mixture is concentrated and chromatographed on silica gel (25 g) with 5% methanol0.5% ammonium hydroxide-methylene chloride; 5 ml fractions are collected. The titled product (Isomer A) is eluted in fractions 69-100 and amounts to 0.009 g. The high resolution FAB mass spectrum has [M + H]⁺ at m/z 831. Theory for C4₅H₆₇N₈O₇: 831.5132. Measured: 831.5112. 4. A solution of the product (Isomer B) prepared in paragraph 2 (0.011 g) in dimethylformamdie (0.13 ml) and tetrahydrofuran (0.72 ml) under a nitrogen atmosphere is treated with 1-hydroxybenzotriazole (0.015 g) and stirred at room temperature for 19 hr. The mixture is concentrated in vacuo to give 0.029 g of a residue which is chromatographed on silica gel (25 g) with 7% methanol-0.5% ammonium hydroxidemethylene chloride; 5.2 ml fractions are collected. The titled product (Isomer B) is eluted In fractions 64-95 and amounts to 0.008 g. The high resolution FAB mass spectrum has [M + H]⁺ at m/z 831. Theory for C₄₅H₆₇N₈O₇: 831.5132. Measured: 831.5151. Preparation 11 (S)-β-[((1,1-Dimethylethoxy)carbonyl)amino]phenylpropanol (Formula D-2) Refer to Chart D. Phenylalaninol (40 g) in tetrahydrofuran (665 ml) is stirred under nitrogen at 0°C in an oven dried 3-neck 3 1 round bottom flask equipped with an addition funnel. Di-t-butyl dicarbonate (63.7 g) In tetrahydrofuran (250 ml) is added over 30 min, after which the ice bath is removed. After stirring 4.25 h, the solvent is removed in vacuo. Recrystallization from carbon tetrachloride/Skelly B affords 59.3 g of the title product as white crystals.

Physical characteristics are as follows: Mp: 96-98°C

IR (cm^-1, mull): 3357, 3319, 2958, 2925, 2870, 2855, 1687, 1670, 1528 , 1454, 1444, 1366, 1315, 1270, 1253, 1185, 1170, 1007, 701.

¹H-NMR (CDCl₃, δ): 7.32, 4.77, 3.82, 3.60, 2.77, 2.32, 1.40. UV λ_max (ε): 207, 238sl sh, 243sl sh, 248, 253, 258, 261sl sh, 264, 268.

Mass spectrum: ions at m/e (relative intensity): 251, 164, 160, 120, 104, 92, 91, 60, 59, 57, 40. Rotation: (α)_D = -24° (0.822, CHC1₃) .

Anal. Found: C, 66.88; H, 8.34; N, 5.73. Preparation 12 (S)-β- [ ( (1,1-Dimethylethoxy)carbonyl)amino]cyclohexanepropanol (Formula D-3) Refer to Chart D. 5% Rhodium on alumina (6.57 g) is added to a solution of the title product of Preparation 11 (57.8 g) in absolute ethanol (79 ml, heating necessary to obtain solution). After this suspension is shaken under 50 lbs hydrogen for 4 days, it is filtered through Celite rinsing with 600 ml ethanol. The filtrate is evaporated in vacuo to afford 61.25 g of the title product. An analytical sample is prepared by Kugelrohr distillation (70-105°C at 1.8 mm Hg) .

Physical characteristics are as follows:

Silica gel TLC: R_f=0.25 in 25% EtOAc/SSB (ninhydrin spray). IR (cm^'1, mull): 3435, 3386, 2930, 2926, 2854, 1692, 1507, 1450, 1393, 1367, 1265, 1247, 1172, 1077, 1061, 1050, 1030, 740, 705. 1H-NMR (CDCI₃, δ): 4.63, 3.80-3.32, 2.40, 1.90-0.70, 1.46. UV λ_maχ (ε) end absorption.

Mass spectrum Found: 257.2001; ions at m/e (relative intensity): 170, 126, 86, 60, 57, 55, 44, 43, 40, 38. Rotation (α)_D - 26° (.698, CHCI³) . Anal. Found: C, 65.02; H, 10.75; N, 5.23.

Preparation 13 (S)- β-(((1,1-Dimethylethoxy) carbonyl)amino)cyclohexanepropanal (Formula D-4) Refer to Chart D. A 2 1 3-neck round bottom flask equipped with an addition funnel, nitrogen inlet, and a thermometer is flame dried and then charged with the title product of Preparation 12 (33.68 g) in DMSO (366 ml) and triethylamine (55 ml). Sulfur trioxide pyridinium complex (62.64 g) in DMSO (366 ml) is added slowly over 70 min, using a cold water bath as necessary to maintain the reaction temperature at 25°C + 2°C After stirring 45 min, the reaction solution is poured over 600 ml of ice and then transferred to a separatory funnel. Extracting once with ether and 2 times with ethyl acetate, washing the combined organic layers 2 times with 10% citric acid, saturated sodium bicarbonate, and brine, and drying over magnesium sulfate affords 32.62 g of crude title product after evaporation of the solvent at 26-30°C This material is used within 3 h without further purification in subsequent steps. Physical characteristics are as follows: Silica gel TLC: Rf-0.35 In 15% ethyl acetate/SSB. 1H-NMR (δ, CDCl₃): 9.68, 4.93, 4.23, 1.86-0.70, 1.45.

Preparation 14 (S-(R*,R*))-(1-(Cyclohexylmethyl-2-hydroxy-4-methyl-3- pentenyl)-carbamic acid, 1,1-dimethylethyl ester (Formula D-5) Refer to Chart D. 1-Bromo-2-methylpropene (17.7 g) is added dropwise to a refluxing mixture of Mg (turnings, 3.05 g), 1,2-dibromo-ethane ( 25 ml), and iodine (0.1 g) in tetrahydrofuran (135 ml). After refluxing until all of the magnesium is in solution (3.8 h), the Grignard reagent is cooled to -10°C The title product of Preparation 13 (13.34 g) in tetrahydrofuran (70 ml) is then slowly added over 55 min. After maintaining the reaction at -10°C for 40 min, the reaction mixture is poured into cold stirring saturated ammonium chloride. After stirring for 20 min, the reaction solution is transferred to a separatory funnel containing ether. Extracting with ether, washing the combined organic layers with saturated sodium bicarbonate and brine, and drying over magnesium sulfate affords 16.7 g of crude title product after evaporation of the solvent. This material is used In the next step without further purification. An analytical sample is prepared by adding a tenth of the crude title product to silica gel (150 g, 15% ethyl acetate/SSB, 35 ml fractions) to afford 0.96 g of the title product. Physical characteristics are as follows:

Silica gel TLC: R_f=0.15 in 5% ethyl acetate/SSB.

IR (mull, cm^-1): 3440, 2977, 2923, 2853, 1717, 1695, 1502, 1449, 1392, 1366, 1248, 1173, 1044, 1022.

¹H-NMR (δ, CDCl₃):5.28, 4.52, 4.28, 3.70, 1.90-0.70, 1.75, 1.70, 1.45.

Mass spectrum (FAB) found: 312.2555 ; ions at m/e: 312, 239 , 238 , 194, 170 , 126 , 95 , 81 , 57 , 55 , 41.

Rotation (α)_D = -24° (0.751, chloroform) . Anal . Found: C , 69 . 17 ; H, 10.43 ; N, 4.41. Preparation 15 (4S - Trans ) -4- (cyclohexylmethyl) -2 , 2 -dimethyl- 5- (2 - methyl-1-propenyl) -3-oxazolidinecarboxylic acid, 1 , 1- dimethylethyl ester (Formula D-6) Refer to Chart D. 2-Methoxypropene (30.2 g) is added to a stirring solution of the title product of Preparation 14 (13.36 g) in methylene chloride (80 ml). Pyridinium-p- toluene sulfonate (0.53 g) is added and the reaction stirred at ambient temperature for 7.5 h. The reaction is quenched with solid sodium bicarbonate. After filtering, the filtrate is evaporated to afford 18.7 g containing the crude title product.

Flash chromatography over silica gel (800 g, 0-2% ethyl acetate/SSB) affords 6.50 g of the title product. Physical characteristics are as follows: Silica gel TLC: R_f=0.5 in 5% ethyl acetate/SSB. IR (mull, cm^-1): 2978, 2926, 2853, 1700, 1449, 1387, 1375, 1365, 1255, 1206, 1178, 1144, 1108, 1087, 1080, 1046, 770.

¹H-NMR (δ, CDCl₃)(2 diastereomers 85/15): 5.31, 4.72, 4.50, 3.67, 1.8-0.7, 1.77, 1.76, 1.57, 1.50. UV λ_max(ε): 228, 236 si sh. Mass spectrum ions at m/e: 351, 336, 280, 267, 236, 194, 166, 126, 111, 57, 55, 41.

Mass spectrum (FAB) ions at m/e: 353, 352, 297, 296, 238. Rotation (α)_D = -2° (0.716, chloroform). Anal. Found: C, 71.79; H, 10.68; N, 3.89. Preparation 16 (4S-(4a, 5b(1S*, 3R*) ) ) 4-(Cyclohexylmethyl)-5-(3-(ethoxycarbonyl)-2,2-dimethylcyclopropyl)-2,2-dimethyl-3- oxazolidinecarboxylic acid, 1,1-dimethylethyl ester (Formula D-7a); (4S-(4a,5b(1R*) ) ) 4-(Cyclohexyl-methyl) 5-(3-(ethoxycarbonyl)-2,2-dimethylcyclopropyl)-2,2- dimethyl-3-oxazolidinecarboxylic acid, 1,1-dimethylethy ester (Formula D-7b); and (4S-(4a,5a)) 4-(Cyclohexylmethyl)-5-(3-(ethoxycarbonyl-2,2-dimethylcyclopropyl)- 2, 2-dimethyl-3-oxazolidinecarboxylic acid, 1,1-dimethylethyl ester (Formula D-7c) Refer to Chart D. The title product of Preparation 15 (13.6 g) and anhydrous copper sulfate (kept in drying oven at 110°C, 78 mg) is stirred at 90°C under vacuum for 40 min. The reaction is then placed under nitrogen and ethyl diazoacetate (20.33 ml) is slowly added over 2 hours with the reaction temperature maintained at 90°C. This material is then allowed to cool to ambient temperature and added to a column.

Flash chromatography over silica gel (800 g, 1.5% EtOAc/SSB, 300 ml fractions) affords 0.98 g of starting material (fractions 23-27). Further elution affords 7.37 g of starting material and cis-diethyl maleonate (fractions 28-46). Further elution affords 1.96 g of the title product D-7a (fractions 86-95).

Physical characteristics are as follows:

Mp: 50-64°C Silica gel TLC: R_f=0.3 in 5% EtOAc/SSB.

IR (cm-¹, mull): 2976, 2952, 2926, 2855, 1729, 1715, 1698, 1456, 1447, 1389, 1377, 1363, 1202, 1185, 1172, 1095, 1055, 769. 1H-NMR (CDCl₃, δ): 4.25, 3.5, 3.20, 1.9-1.0, 1.45, 1.28, 1.25, 1.21. 1³C-NMR (CDCl₃, ppm): 171.08, 151.22, 93, 80.5, 79.1, 60.4, 59.8, 41.1, 37.0, 35.1, 33.8, 31.9, 31.6, 28.0, 26.0, 25.9, 25.6, 21.4, 20.0, 13.8.

UV λ_maχ(ε): 276 sh.

Mass spectrum (FAB) found: 438.3189; ions at m/e: 438, 392, 380, 338, 324, 196, 95, 81, 58, 57, 55, 41, 29.

Rotation (α)_D = 0 (0.95, chloroform).

Anal. Found: C, 68.95; H, 10.14; N, 3.04.

Further elution affords 10.86 g of a mixture of the title products of formulas D-7a and D-7b, and trans-dlethyl maleonate (fractions 96-142). Further elution affords 1.20 g of the title product of formula D-7c (fractions 146-159), m.p. 110-124°C

Physical characteristics are as follows:

Silica gel TLC: R_f=0.25 in 5% ethyl acetate/SSB.

IR (mull, cm^-1): 2983, 2955, 2926, 2916, 2867, 2854, 1723, 1686, 1463, 1456, 1395, 1389, 1378, 1373, 1366, 1253, 1180, 1175, 1157, 1116, 1089, 1022, 844, 770. 1H-NMR (δ, CDCI₃): 4.15, 3.95, 3.60, 2.2-0.7, 1.50.

¹³C-NMR (CDCI₃, ppm) (2 compounds): (Major diastereomer) : 171.02, 151.2, 91.71, 79.01, 76.90, 59.84, 56.25, 38.1, 33.62, 32.94, 31.88, 30.48, 28.0.8, 27.65, 26.73, 26.07, 25.94, 25.17, 23.21, 21.52, 20.23, 13.84; (Minor diastereomer) : 171.02, 51.6, 91.2, 79.11, 76.76, 59.85, 56.25, 37.6, 33.62, 32.94, 31.88, 30.48, 28.08, 27.65, 26.73, 26.07, 25.94, 25.17, 23.21, 21.52, 20.23, 13.84.

Mass spectrum (FAB) M+1 Found: 438.3231; Ions at m/e: 438, 380, 338, 324, 280, 95, 83, 73, 58, 57, 43, 41, 29.

Rotation (α)_D = +4° (0.983, chloroform).

Flash chromatography of the 10.86 g of mixed fractions over silica gel (800 g, 2% acetone/SSB, 300 ml fractions) affords 6.43 g of mixed title products of formulas D-7a and D-7b (fractions 18-21) and 0.32 g of title product of formula D-7b (fractions 22-24). Physical characteristics are as follows: Silica gel TLC: R_f=0. 27 in 5% ethyl acetate/SSB. IR (mull, cm^-1): 2927, 1726, 1700, 1449, 1387, 1366, 1256, 1251, 1203, 1179, 1176, 1149, 1117, 1097, 1077, 854, 849, 769.

¹H-NMR ( 5 , CDCl₃): 4.09, 3.95, 3.55, 1.90-0.07, 1.47. 1³C-NMR (CDCI₃, ppm) (2 compounds): (Major diastereomer): 170.89, 150.95, 92.7, 78.8, 78.0, 60.96, 59.44, 40.1, 36.77, 34.72, 33.82, 31.47, 30.82, 27.80, 27.38, 25.76, 25.51, 20.84, 20.09, 13.69; (Minor diastereomer): 170.43, 150.95, 92.7, 78.8, 78.0, 60.77, 59.13, 40.1, 36.77, 34.46, 33.82, 31.47, 30.82, 27.80, 27.38, 25.76, 25.51, 20.84,, 20.09, 13.32.

UV λ_maχ(ε): 280 sl sh. Mass spectrum (FAB) M+1 Found: 438.3231; ions at m/e: 438, 422, 380, 338, 324, 196, 95, 73, 58, 57, 55, 41, 29.

Rotation (α)_D = -19° (0.522, chloroform). Preparation 17 (1R-(1α, 2β (1R*,2R*))) 3- (3-Cyclohexyl-2-(((1,1-dimethylethoxy) carbonyl) amino) -1-hydroxypropyl)-2,2-dimethylcyclopropanecarboxylic acid, ethyl ester (Formula D-8)

Refer to Chart D. Tosic acid (0.2 g) is added to a stirring solution of the title product of formula D-7b of Preparation 16 (218 mg) in methanol, 6.7 ml at 60°C After 1 h, the reaction is quenched with solid sodium bicarbonate. After evaporation the residue is added to a column.

Flash chromatography over silica gel (15 g, 15% ethyl acetate/SSB) affords 58 mg of the title product.

Physical characteristics are as follows: Silica gel TLC: R_f=0.3 in 25% ethyl acetate/SSB. 1H-NMR (δ, CDCI₃): 4.83, 4.01, 3.64, 3.13, 1.80-0.60, 1.32, 1.15, 1.11, 1.06.

¹³C-NMR (CDCI₃, ppm): 172.37, 155.80, 78.83, 73.40, 60.24, 51.49, 39.68, 36.10, 34.00, 33.60, 32.61, 31.74, 28.15, 27.17, 26.32, 26.12, 25.93, 21.46, 20.25, 14.06. Mass spectrum (FAB) M+1 Found: 398.2912; ions at m/e: (relative intensity) 398, 298, 280, 170, 126, 95, 83, 57, 55, 41, 29.

Anal. Found: C, 66.22; H, 10.09; N, 3.40. Preparation 18 (1R- ( 1a, 2β(1R*, 2R*) ) )3- (3-Cyclohexyl-2-(((1,1-dimethyl ethoxy) carbonyl) amino ) - 1 -hydroxypropyl) - 2 , 2 -dImethylcyclopropanecarboxylic acid, ethyl ester (Formula D-8)

Refer to Chart D .

Tosic acid (0.2 g) Is added to a stirring solution of the title product of formula D- 7b of Preparation 16 (218 mg) in methanol (6 . 7 ml) at ambient temperature . After 4.5 h, the reaction is quenched with solid sodium bicarbonate . After evaporation, the residue Is added to a column.

Flash chromatography over silica gel (15 g, 15% ethyl acetate/SSB) affords 75 mg of the title product which is Identical by silica gel

TLC, ¹H-NMR, and ¹³C-NMR to previously prepared material. Further elution affords 31 mg of the title product containing a small amount of a second diastereomeric ethyl ester.

Physical characteristics are as follows : Silica gel TLC for the second diastereomer: R_f=0.27 in 25% ethyl acetate/SSB. 1H-NMR (CDCI₃, δ) looks similar to that of the title product. 1³C-NMR (CDCI₃, ppm): 172.37, 155.80, 78.83, 73.40, 60.24, 51.49, 39.68, 36.70, 34.00, 33.60, 32.61, 31.74, 28.15, 27.17, 26.32, 26.12, 25.93, 21.61, 20.48, 14.06.

Preparation 19 (1R- (1α , 2β(1S* , 2S*) ) ) 3- (3-Cyclohexyl-2- ( ( (1 , 1-dlmethylethoxy)carbonyl) amino) -1-hydroxypropyl)-2,2-dlmethylcyclopropanecarboxylic acid, ethyl ester. Refer to

Chart D. Tosic acid (0.2 g) is added to a stirring solution of the title product of formula D-7a of Preparation 16 (1.09 g) in methanol (33 ml).

After 7 h, the reaction is quenched with solid sodium bicarbonate.

After evaporation, the residue is added to a column.

Flash chromatography over silica gel (80 g, 15% ethyl acetate/SSB) affords 0.65 g of the title product.

Physical characteristics are as follows: Silica gel TLC: R_f=0.3 in 25% ethyl acetate/SSB. 1H-NMR (CDCI₃, δ): 4.64, 4.12, 3.70, 3.25, 2.58, 1.90-0.70, 1.44, 1.38, 1.26, 1.27, 1.25. 1³C-NMR (CDCI₃, ppm): 171.54, 156.38, 79.43, 74.53, 60.33, 52.71, 39.20, 35.64, 34.17, 34.10, 32.54, 31.95, 28.35, 27.42, 26.52, 26.32, 26.09, 21.60, 20.89, 14.39.

Mass spectrum (FAB) M+1 Found: 398.2910; ions at m/e (relative intensity) 398, 342, 324, 298, 170, 126, 95, 83, 81, 57, 55, 41, 29.

Anal. Found: C, 66.47; H, 10.09; N, 3.47. Preparation 20 ( 1S - ( 1β2β) )3- (3-Cyclohexyl-2- ( ( (1,1-dimethylethoxy)carbonyl) amino)-1-hydroxypropyl)-2,2-dimethylcyclopropanecarboxylic acid, ethyl ester. Refer to Chart D.

Tosic acid (25 mg) is added to a stirring solution of the title product of formula D-7c of Preparation 16 (0.14 g) in methanol (4.3 ml). After 5 h, the reaction is quenched with solid sodium bicarbonate. After evaporation, the residue is added to a column. Flash chromatography over silica gel (15 g, 15% ethyl acetate/SSB) affords 50 mg of the title product.

Physical characteristics are as follows: Silica gel TLC: R_f=0.3 in 25% ethyl acetate/SSB.

¹H-NMR (CDCl₃, δ): 4.75, 4.12, 3.75, 3.40, 2.80, 2.00-0.60, 1.60, 1.48.

Preparation 21 [1S- [ 1α,2β(1R*, 2R*) ] ] 3 [3-Cyclohexyl-2- [ [ (1,1-dimethylethoxy)carbonyl] amino]-1-hydroxypropyl]-2,2-dimethylcyclopropanecarboxylic acid (Formula D-9) Refer to Chart D. Lithium hydroxide (2N, 9 ml) is added to a solution of the title product of Preparation 17 (0.23 g) in methanol (9 ml). After stirring at 40° C for 6 h, the reaction solution is transferred to a separatory funnel with 2N hydrochloric acid/ether. Extracting 3 times with ether, washing the combined organic layers with brine, and drying over magnesium sulfate affords 0.18 g of the title product after evaporation of the solvent.

Physical characteristics are as follows:

Silica gel TLC: Rf-0.25 in 2% acetic acid, 50% ethyl acetate/SSB. 1H-NMR (δ, CDCI₃): 7.3, 5.60, 5.02, 3.75, 3.63, 3.25, 1.90-0.80, 1.44.

¹³C-NMR (CDCI3, ppm): 177.3, 156.1, 79.1, 73.8, 51.8, 39.9, 37.3, 34.2, 33.9, 32.9, 32.0, 28.4, 26.6, 26.3, 26.2, 21.8, 20.6.

Mass spectrum ions at m/e: 226, 170, 126, 97, 83, 81, 69, 67, 59, 57, 55. Preparation 22 [ 1R- [ 1α,2β(1S*, 2S*) ] ] 3 [3-Cyclohexyl- 2- [ [ (1,1-dimethylethoxy)carbonyl] amino] -1-hydroxypropyl] -2,2-dimethylcyclopropanecarboxylic acid. Refer to Chart D. Lithium hydroxide (2N, 16 ml) is added to a solution of the title product of Preparation 19 (0.4 g) in methanol (16 ml). After stirring at 40°C for 5 h and at ambient temperature for 17 h, the reaction solution is transferred to a separatory funnel with 2N hydrochloric acid/ether. Extracting three times with ether, washing the combined organic layers with brine, and drying over magnesium sulfate afford 0.32 g of the title product after evaporation of the solvent. Physical characteristics are as follows:

Silica gel TLC: R_f=0.45 in 0.5% acetic acid, 50% ethyl acetate/-SSB. IR (cm^-1, mull): 3311, 2956, 2916, 2868, 2855, 1723, 1695, 1460, 1456, 1378, 1366, 1171.

¹H-NMR (CDCl₃, δ): 6.8, 3.75, 3.2, 1.90-0,70, 1.40. 1³C-NMR (CDCI₃, ppm): 176.1, 82.8, 74.1, 56.2, 43.0, 35.4, 34.2, 33.2, 32.8, 32.4, 31.6, 28.2, 26.2, 26.0, 25.9, 21.4, 20.2. Mass spectrum (FAB) Found: 370.2578; ions at m/e (relative intensity) 370, 314, 296, 270, 250, 170, 154, 126, 55, 41. Preparation 23 [ 1S - [ 1α, 2β(1R*, 2R*) ] ] 3 [3-Cyclohexyl-2- [ [ (1,1-dimethylethoxy) carbonyl] amino]-1-hydroxy-propyl]-2,2-dimethylcyclopropanecarboxylic acid, L-isoleucyl-2-pyridylmethylamide (Formula D-10) Refer to Chart D.

The title product of Preparation 21 (185 mg), isoleucyl-2-pyridylmethylamide (110.6 mg) , and dlethyl cyanophosphonate (95 mg) are dissolved in dimethylformamide (4 ml) at 0°C. Triethylamine (55.3 mg) is added and the ice bath removed. After 2.5 h, the reaction solution is diluted with 60 ml of 2/1 ethyl acetate/benzene and transferred to a separatory funnel. Washing with water, saturated sodium bicarbonate, and brine, and drying over magnesium sulfate affords 303 mg of crude title product after evaporation of the solvent.

Flash chromatography over silica gel (30 g, 1-3% methanol/ethyl acetate, 5 ml fractions) affords 209 mg of the title product (fractions 51-67).

Physical characteristics are as follows: Silica gel TLC: R_f=0.5 in 5% methanol/ethyl acetate. HPLC: 28.2% RT-11.65 min and 67.6% RT-14.6 min. [Synchrom RP-18, 1.5 ml/min, 17% B 2 min., linear gradient 17% to 100% B; A - 10% CH₃CN-H₂O, 0.2% TFA; B - 70% CH₃CN-H₂O, 0.29. TFA. ]

IR (mull, cm^-1): 3284, 2967, 2862, 2850, 1715, 1668, 1636, 1537, 1501, 1462, 1377, 1365, 1173. ¹H-NMR (δ, CDCI₃): 8.46, 8.18, 7.58, 7.28, 7.15, 5.12, 4.6-4.2, 3.71, 3.14, 1.9-0.81, 1.40.

¹³C-NMR (CDCI₃, ppm): 172.4, 171.6, 157.1, 155.9, 148.8, 137.1,

122.7, 122.5, 78.7, 73.4, 58.1, 51.8, 44.6, 40.3, 37.3, 36.4, 34.3, 34.0, 33.9, 32.9, 28.4, 26.6, 26.2, 25.9, 24.9, 21.7, 20.6, 15.5, 11.1,

Mass spectrum (FAB) Found: 573.4044; ions at m/e: 574, 573, 517,

499, 473, 409, 248, 109, 95, 93, 86, 57, 55.

Preparation 24 [1R- [ 1α, 2β(1S*, 2S*) ] ] 3 [3 -Cyclohexyl-2- [ [ (1, 1-dimethylethoxy)carbonyl] amino] -1-hydroxypropyl]-2,2-dimethylcyclopropanecarboxylic acid, L-isoleucyl-2-pyridylmethylamide. Refer to Chart D. The title product of Preparation 22 (95 mg), isoleucyl-2-pyridylmethylamide (56.8 mg), and diethyl cyanophosphonate (48.8 mg) are dissolved in dimethylformamide (2 ml) at 0°C Triethylamine (28.4 mg, .039 ml) is added and the ice bath removed. After 2.5 h, the reaction solution is diluted with 30 ml of 2/1 ethyl acetate/benzene and transferred to a separatory funnel. Washing with water, saturated sodium bicarbonate, and brine, and drying over magnesium sulfate affords 177 mg of crude title product after evaporation of the solvent. Flash chromatography over silica gel (18 g, 1-3% methanol/ethyl acetate, 5 ml fractions) affords 92.8 mg of the title product (frac- tions 16-25).

Physical characteristics are as follows: Mp 92-98°C. Silica gel TLC: R_f=0.5 in 5% methanol/ethyl acetate.

HPLC: 28.2% RT 11.65 min and 67.6% RT 14.6 min [synchorm RP-18, 1.5 ml/min, 17% B 2 min., linear gradient 17% to 100% B (A - 10% CH₃CN-H₂O, 0.2% TFA; B = 70% CH₃CN-H₂O, 0.2% TFA)].

IR (KBr, c^m-1): 3298, 2965, 2926, 1745, 1681, 1646, 1529, 1391, 1380, 1366, 1242, 1172. 1H-NMR (CDCI₃, δ): 8.51, 7.63, 7.30-7.00, 4.9, 4.55, 4.42, 3.76, 3.2, 2.0-0.7, 1.42.

¹³C-NMR (CDCI₃, ppm): 171.8, 170.5 and 169.7, 156.9, 156.3,

149.0, 136.8 and 136.7, 122.4 and 122.2, 121.8 and 121.6, 83.0 and 74.9, 79.5, 58.2 and 58.0, 56.3 and 53.8, 44.5 and 43.4, 39.2, 37.4 and

36.7, 34.2, 33.8 and 33.6, 32.8 and 32.7,- 28.4, 26.5, 25.1, 24.7, 21.5,

20.1, 15.6 and 14.2, 11.2 and 11.0.

Mass spectrum (FAB) Found: 573.4017; ions at m/e (relative intensity) 573, 517, 499, 473, 409, 248, 109, 95, 93, 86, 69, 57, 55,

41.

Preparation 25 [1S-[ 1α, 2β(1R*,2R*) ]] 3[ 3 -Cyclohexyl-2-[(amino)-1- hydroxypropyl]-2,2-dimethylcyclopropanecarboxylic acid, L-isoleucyl-2-pyridylmethylamide (Formula D-11) Refer to

Chart D. A solution of the title product of Preparation 23 (0.2 g) in methylene chloride (8 ml) and trifluoroacetlc acid (8 ml), is placed at -20°C After 19 h, the reaction solution is transferred to a separatory funnel containing saturated sodium bicarbonate. Extracting 2 times with methylene chloride and drying over magnesium sulfate affords 139.2 mg of the title product.

Physical characteristics are as follows: 1H-NMR (δ, CDC1₃): 8.49, 8.34, 7.78, 7.62, 7.27, 7.15, 5.23, 4.45, 3.16, 3.00, 1.91, 1.80-0.70.

¹³C-NMR (CDCI₃, ppm): 172.7, 172.1, 157.2, 148.8, 137.0, 122.4,

122.1, 72.4, 58.4, 54.4, 44.6, 39.1, 37.0, 35.5, 34.1, 33.6, 32.7,

32.5, 27.2, 26.4, 26.1, 25.9, 24.9, 22.5, 21.0, 20.6, 15.6, 11.2.

Preparation 26 [1S- [1α,2β(1R*, 2R*) ] ]3 [3-Cyclohexyl-2- [ [L-Boc-His(Ts) (a- mino) -1-hydroxypropyl]-2,2-dimethylcycloproρanecarboxylic acid, -L-isdleucyl-2-pyr.idylmethylamide (Formula

D-12) Refer to Chart D.

The title product of Preparation 25 (139 mg), boc-his(ts) (153.7 mg) , and diethyl cyanophosphonate (77.3 mg) are dissolved in dimethylformamide (1.2 ml) at 0°C Triethylamine (61.8 mg) is added and the ice bath removed. After 18.5 h, the reaction solution is diluted with

35 ml of 2/1 ethyl acetate/benzene and transferred to a separatory funnel. Washing with water, saturated sodium bicarbonate, and brine, and drying over magnesium sulfate affords 234 mg of crude title product after evaporation of the solvent.

Flash chromatography over silica gel (25 g, 5% methanol/ethyl acetate, 2 ml fractions) affords 159.2 mg of the title product (fractions 33-40).

Physical characteristics are as follows: Silica gel TLC: Rf-0.15 in 5% methanol/methylene chloride. 1H-NMR (δ, CDCI₃): 8.48, 8.10, 8.01, 7.79, 7.66, 7.54, 7.29, 7.30, 7.18, 7.13, 5.48, 4.53, 3.95, 3.14, 2.96, 2.77, 2.41, 2.05-0.70, 1.38. ¹³C-NMR (CDCI₃, ppm): 172.7, 171.9, 171.5, 156.3, 155.5, 148.9,

146.0, 140.8, 137.0, 136.7, 135.0, 130.3, 127.4, 122.5, 122.3, 114.8,

79.3, 73.2, 58.4, 54.0, 53.5, 50.4, 44.5, 39.7, 37.0, 34.2, 34.0, 33.8,

32.6, 31.8, 28.3, 26.6, 26.2, 26.0, 25.5, 24.8, 21.8, 21.7, 20.6, 15.7, 11.0.

Preparation 27 [1S- [ 1α, 2β(1R*.2R*) ] ] 3 [3-Cyclohexyl-2-[[L-His(Ts) (amino)-1-hydroxypropyl]-2,2-dimethyleyclopropanecarboxylic acid, -L-isoleucyl-2-pyridylmethylamide (Formula D-13) Refer to Chart D. A solution of the title product of Preparation 26 (159.2 g) in methylene chloride (4 ml) and trifluoroacetic acid (4 ml) is placed at -20°C After 3 h, the reaction solution is transferred to a separatory funnel containing saturated sodium bicarbonate. Extracting 2 times with methylene chloride and drying over magnesium sulfate affords 133.1 mg of the title product.

Physical characteristics are as follows:

¹-H-NMR (δ, CDCI₃): 8.47, 8.04, 7.94, 7.79, 7.64, 7.55, 7.33, 7.28, 7.18, 7.15, 4.55-4.40, 3.96, 3.70, 3.12, 2.42, 2.0-0.80.

¹³C-NMR (CDCI₃, ppm): 173.4, 172.4, 171.9, 156.4, 148.9, 146.2, 141.8, 137.0, 136.4, 135.0, 130.4, 127.4, 122.4, 122.2, 114.8, 72.5, 58.3, 55.1, 53.5, 50.6, 44.5, 39.5, 37.1, 34.3, 33.9, 33.6, 32.8, 30.9, 26.5, 26.2, 26.1, 25.8, 25.7, 24.8, 21.7, 20.7, 15.6, 11.0. Preparation 28 [1S-[ 1α, 2β(1R*, 2R*) ] ] 3 [3-Cyclohexyl-2-[L-Boc-Phe-L- His (Ts ) (amino)-1-hydroxypropyl]-2,2-dimethylcyclopro- panecarboxylic acid, -L-isoleucyl-2-pyridylmethylamide

(Formula D-14) Refer to Chart D.

The title product of Preparation 27 (65 mg) , L-boc-phe (29.3 mg), and diethyl cyanophosphonate (22.4 mg) are dissolved in methylene chloride (0.35 ml) at 0°C under nitrogen. N,N-diisopropylamine (22.5 mg) is added and the ice bath removed. After 16.5 h, the reaction solution is evaporated in vacuo to afford crude title product.

Flash chromatography over silica gel (10 g, 3% methanol/methylene chloride, 1 ml fractions) affords 71.6 mg of the title product (fractions 35-84). Physical characteristics are as follows:

Silica gel TLC: R_f=0.40 in 5% methanol/methylene chloride. 1H-NMR (δ, CDCI₃): 8.49, 7.96, 7.79, 7.65, 7.30-7.11, 4.60-4.30, 3.93, 3.09, 2.89, 2.39, 1.60-0.83, 1.37. ¹³C-NMR (CDCI₃, ppm): 172.23, 171.55, 171.47, 170.12, 156.88,

148.90, 146.26, 140.27, 137.02, 136.45, 134.93, 134.78, 130.39, 129.24,

128.65, 127.46, 127.01, 122.37, 115.08, 80.43, 79.35, 58.29, 56.28,

53.25, 51.78, 44.57, 39.65, 39.15, 37.98, 37.26, 33.89, 32.65, 29.68, 28.29, 26.55, 25.99, 24.88, 21.67, 20.70, 15.65, 11.21.

Preparation 29 [1S- [1α, 2β(1R*, 2R*) ] ]3[3-Cyclohexyl-2-[(Z)- (2-phenyl-1- (Boc)aminocyclopropane-1-carboxyl)-L-His(Ts) (amino) -1- hydroxypropyl]-2,2-dimethylcyclopropanecarboxylic acid, -L-isoleucyl-2-pyridylmethylamide (Formula E-2) Refer to Chart E.

The title product of Preparation 27 (65 mg), (Z) -2-phenyl-1-(boc)amino-cyclopropane-1-carboxylic acid (30.7 mg), and diethyl cyanophosphonate (22.4 mg) are dissolved in methylene chloride (0.35 ml) at 0°C under nitrogen. N,N-diisopropylamine (22.5 mg) is added and the ice bath removed. After 19.5 h, the reaction solution is evaporated in vacuo to afford crude title product.

Flash chromatography over silica gel (10 g, 3% methanol/methylene chloride, 2 ml fractions) affords 42.3 mg of the title product (first diastereomer) (fractions 20-38). Physical characteristics are as follows:

Silica gel TLC: R_f=0.35 in 5% methanol/methylene chloride. 1H-NMR (δ, CDCI₃): 8.47, 7.98-7.61, 7.37-6.90, 5.62, 4.75, 4.54-4.44, 3.96, 3.25-2.80, 2.41, 1.91, 1.60-0.80.

¹³C-NMR (CDCI₃, ppm): 172.11, 171.74, 171.3, 170.8, 156.2, 148.94, 146, 140.69, 136.91, 134.79, 130.42, 128.60, 127.47, 122.31, 115, 80, 73.20, 5.51, 37.01, 34.32, 33.92, 32.67, 28.29, 26.58, 26.00, 24.84, 21.85, 21.72, 15.61, 11.25.

Further elution affords 22. 9 mg of the title product (second diastereomer) (fractions 42- 60) . Physical characteristics are as follows :

Silica gel TLC : R_f=0.30 In 5% methanol/methylene chloride . 1H-NMR (δ , CDCI₃) : 8.47 , 7 .90-7.61 , 7. 37 -7.00 , 6 . 62, 5.05 , 4.75 -4.08 , 4.00 , 3 .40-3.10 , 2.43 , 1.91 , 1.60-0. 80.

Example 3 [ 1S - [ 1α , 2β(lR* , 2R*) ] ] 3 [3 -Cyclohexyl- 2- [ [L-Boc-Phe-L- His (amino) - 1 -hydroxypropyl ]- 2 , 2- dimethylcyclopropanecar- boxylic acid, -L-isoleucyl-2-pyridylmethylamide (Formula D-15) Refer to Chart D . A solution of the the title product of Preparation 28 (71.6 mg) and 1-hydroxybenzotriazole (70 mg) in methanol (2.5 ml) is stirred at ambient temperature. After 17.5 h, the solution is evaporated in vacuo and the residue is submitted to flash chromatography over silica gel (12 g, 5% methanol (saturated with ammonia/aethylene chloride, 3 ml fractions) to afford 26.7 mg of the title product (fractions 51-57). Physical characteristics are as follows:

Silica gel TLC: R_f=0.5 in 10% methanol (ammonia)/methylene chloride.

Mass spectrum (FAB) Found: 857.5289; ions at m/e: 858, 857, 12 110, 109, 95, 83, 57, 55, 41.

Example 4 [1S- [1α, 2β(1R*, 2R*) ] ] 3 [3-Cyclohexyl-2-[[(Z)-(2-phenyl- (boc) aminocyclopropane-1-carboxyl)-L-His(amino)-1- hydroxypropyl] -2,2-dimethylcyclopropanecarboxylic acid,

-L-isoleucyl-2-pyridylmethylamide (Formula E-3) Refer to Chart E.

A solution of the title product of Preparation 29 (the more polar epimer, 22.9 mg) and 1-hydroxybenzotriazole (22 mg) in methanol (0.8 ml) is stirred at ambient temperature. After 22 h, the solution is evaporated in vacuo and the residue is submitted to flash chromatography over silica gel (2.5 g, 5% methanol (saturated with ammonia/methylene chloride, 3 ml fractions) to afford 2.7 mg of the title product (fractions 18-25).

Physical characteristics are as follows:

Silica gel TLC: R_f=0.4 in 10% methanol (ammonia)/methylene chloride.

Mass spectrum (FAB) Found: 869.5255; ions at m/e: 869, 110, 109, 95, 83, 82, 69, 57, 55, 41.

, ^

C

B

H

Claims

1. A renin inhibitory peptide having a non-cleavable transition state insert corresponding to the 10, 11-position of a renin substrate (angiotensinogen) and having:

(a) at least one cyclopropyl amino acid of the formula XL_2b

in place of an amino acid residue corresponding to position 6, 8, 9, 12, 13 or 14 of the renin substrate; and/or

(b) a cycloalkyl amino acid non-cleavable transition state insert of the formula XL_2c

in place of an amino acid residue corresponding to position 10 and 11 of the renin substrate; wherein R₅₀ is

(a) hydrogen,

(b) C₁-C₅alkyl, (c) -(CH₂)_p-aryl,

(d) -(CH₂)_p-Het,

(e) -(CH₂)p-C₃-C₇cycloalkyl, or

(f) 1- or 2-adamantyl; wherein R₆₀ and R₆₁ are the same or different and are (a) hydrogen,

(b) Chalky!,

(c) -(CH₂)_p-aryl,

(d) -(CH₂)_p-Het,

(e) -(CH₂)p-C₃-C₇cycloalkyl, or (f) 1- or 2-adamantyl; or wherein R₆₀ and R₆₁ taken together form a spirocycle of the formula II

wherein R₇₀ is

(a) -CHR₈₀,

(b) -O-,

(c) -S-,

(d) -SO-,

(e) -SO₂-, or

(f) -NR₈₁; wherein R₈₀ and R₈₁ are the same or different and are

(a) hydrogen,

(b) C₁-C₅alkyl,

(c) -(CH₂)_p-aryl,

(d) -(CH₂)_p-Het,

(e) -(CH₂)_p-C₃-C₇ cycloalkyl, or

(f) 1- or 2-adamantyl; wherein R₉₀ is

(a) hydrogen, or

(b) C₁-C₅alkyl; wherein R₁₀₀ is

(a) hydrogen,

(b) C₁-C₅alkyl ,

(c) aryl,

(d) C₃-C₇cycloalkyl,

(e) -Het,

(f) C₁-C₃alkoxy, or

(g) C₁-C₃alkylthio; wherein R₁₁₀ is

(a) hydrogen,

(b) C₁-C₅alkyl,

(c) -(CH₂)_p-aryl, or

(d) halogen; wherein R₁₁₁ is

(a) hydrogen,

(b) C₁-C₅alkyl,

(c) -(CH₂)_p-aryl, or

(d) halogen; wherein p is zero to two, inclusive; wherein r is zero to three, inclusive; wherein t is zero to three, inclusive; wherein aryl is phenyl or naphthyl substituted by zero to 3 of the following: (a) C₁-C₃alkyl,

(b) hydroxy,

(c) C₁-C₃alkoxy,

(d) halo,

(e) amino, (f) mono- or di-C₁-C₃alkylamlno,

(g) -CHO, (h) -COOH, (I) COOR₂₆, (j) CONHR₂₆ , (k) nitro,

(1) mercapto, (m) C₁-C₃alkylthio, (n) C₁-C₃alkylsulfinyl, (o) C₁-C₃alkylsulfonyl, (p) -N(R₄)-C₁-C₃alkylsulfonyl,

(q) SO₃H, (r) SO₂NH₂, (s) -CN, or (t) -CH₂NH₂; wherein -Het is a 5- or 6-membered saturated or unsaturated ring containing from one to three heteroatoms selected from the group consisting of nitrogen, oxygen, and sulfur; and including any bicycllc group in which any of the above heterocyclic rings is fused to a benzene ring, which heterocyclic moiety is substituted with zero to 3 of the following:

(i) C₁-C₆alkyl, (ii) hydroxy, (iii) trifluoromethyl, (iv) C₁-C₄alkoxy, (v) halo,

(vi) aryl, (vii) aryl C₁-C₄alkyl-, (viii) amino, (ix) mono- or di- (C₁-C₄alkyl)amino, and (x) C₁-C₅alkanoyl; wherein R₄ at each occurrence is the same or different and is (a) hydrogen, (b) C₁-C₅alkyl,

(c) -(CH₂)_p-aryl,

(d) -(CH₂)_p-Het,

(e) -(CH₂)p-C₃-C₇ cycloalkyl, or

(f) 1- or 2-adamantyl; wherein R₂₆ is

(a) hydrogen,

(b) C₁-C₃alkyl, or

(c) phenyl-C₁-C₃alkyl.

2. In a renin inhibitory peptide having a non-cleavable transition state insert corresponding to the 10, 11-position of a renin substrate (angiotensinogen), the improvement which comprises inclusion in the renin inhibitory peptide of:

(a) at least one cyclopropyl amino acid of the formula X_L2b

in place of an amino acid residue corresponding to position 6, 8, 9,

12, 13 or 14 of the renin substrate; and/or

(b) a cycloalkyl amino acid non-cleavable transition state insert of the formula XL_2c

in place of an amino acid residue corresponding to position 10 and 11 of the renin substrate; wherein R₅₀ is

(a) hydrogen,

(b) C₁-C₅alkyl,

(c) -(CH₂)_p-aryl, (d) - (CH₂)_p-Het,

(e) - (CH₂)_p-C₃-C₇ cycloalkyl , or

(f) 1- or 2-adamantyl ; wherein R₆₀ and R₆₁ are the same or different and are (a) hydrogen,

(b) C₁-C₅alkyl ,

(c) - (CH₂)_p- aryl ,

(d) - (CH₂)_p-Het,

(e) - (CH₂)_p-C₃-C₇cycloalkyl , or (f) 1- or 2-adamantyl ; erein R₆₀ and R₆₁ taken together form a spirocycle of the formula

wherein R₇₀ is

(a) -CHR₈₀

(b) -O-,

(c) -S-,

(d) -SO-,

(e) -sO₂-, or

(f) -NR₈₁; wherein R₈₀ and R₈₁ are the same or different and are (a) hydrogen, (b) C₁-C₅alkyl,

(c) -(CH₂)_p-aryl,

(d) -(CH₂)_p-Het,

(e) -(CH₂)_p-C₃-C₇ cycloalkyl, or

(f) 1- or 2-adamantyl; wherein R₉₀ is

(a) hydrogen, or

(b) C₁-C₅alkyl; wherein R₁₀₀ is

(a) hydrogen, (b) C₁-C₅alkyl,

(c) aryl,

(d) C₃-C₇cycloalkyl,

(e) -Het, (f) C₁-C₃alkoxy, or

(g) C₁-C₃alkylthio; wherein R₁₁₀ is

(a) hydrogen, (b) C₁-C₅alkyl,

(c) -(CH₂)_p-aryl, or

(d) halogen; wherein R₁₁₁ is

(a) hydrogen, (b) C₁-C₅alkyl,

(c) -(CH₂)_p-aryl, or

(d) halogen; wherein p is zero to two, inclusive; wherein r is zero to three, inclusive; wherein t is zero to three, inclusive; wherein aryl is phenyl or naphthyl substituted by zero to three ofhe following:

(a) C₁-C₃alkyl,

(b) hydroxy, (c) C₁-C₃alkoxy,

(d) halo,

(e) amino,

(f) mono- or di-C₁-C₃alkylamino,

(g) -CHO, (h) -COOH,

(i) COOR₂₆,

(j) CONHR₂₆ ,

(k) nitro,

(l) mercapto, (m) C₁-C₃alkylthio,

(n) C₁-C₃alkylsulfinyl,

(o) C₁-C₃alkylsulfonyl,

(p) -N(R₄)-C₁-C₃alkylsulfonyl,

(q) SO₃H, (r) SO2NH₂,

(s) -CN, or

(t) -CH₂N_H2 ; wherein -Het is a 5 - or 6-membered saturated or unsaturated ring containing from one to three heteroatoms selected from the group consisting of nitrogen, oxygen, and sulfur; and including any bicyclic group in which any of the above heterocyclic rings is fused to a benzene ring, which heterocyclic moiety is substituted with zero to three of the following:

(i) C₁-C₆alkyl, (ii) hydroxy, (iii) trifluoromethyl, (iv) C₁-C₄alkoxy, (v) halo,

(vi) aryl, (vii) aryl C₁-C₄alkyl-, (viii) amino,

(ix) mono- or di- (C₁-C₄alkyl)amino, and (x) C₁-C₅alkanoyl; wherein R₄ at each occurrence is the same or different and is

(a) hydrogen,

(b) C₁-C₅alkyl,

(c) -(CH₂)_p-aryl, (d) -(CH₂)_p-Het,

(e) -(CH₂)_p-C₃-C₇ cycloalkyl, or

(f) 1- or 2-adamantyl; wherein R₂₆ is

(a) hydrogen, (b) C₁-C₃alkyl, or

(c) phenyl-C₁-C₃alkyl.

3. The renin inhibitory peptide of claim 1 wherein the cyclopropyl amino acid of the formula XL_2b Is at the N- terminus of the peptide.

4. The renin inhibitory peptide of claim 2 wherein the cyclopropyl amino acid of the formula XL_2b is at the N-terminus of the peptide.

5. The renin inhibitory peptide of claim 1 of the formula I X-Ag-B₇-C₈-D₉-E₁₀-F₁₁-G₁₂-H₁₃-I₁₄-Z I wherein X is

(a) hydrogen,

(b) C₁-C₅alkyl, (c) R₅-O-CH₂-C(O)-,

(d) R₅-CH₂-O-C(O)-,

(e) R₅-O-C(O)-,

(f) R₅-(CH₂)_n-C(O)-, (g) R₄N(R₄)-(CH₂)_n-C(O),

(h) R₅-SO₂-(CH₂)_q-C(O)-,

(i) R₅-SO₂-(CH₂)_q-O-C(O)-, or

(j) R₆-(CH₂)_i-C(O)-; wherein A₆ is absent or a divalent moiety of the formula XL₁ , XL₂ , XL_2a, or XL_2b;

wherein B₇ is absent or a divalent moiety of the formula XL_b;

wherein C₈ is absent or a divalent moiety of the formula XL₁, XL₂ , XL_2a, or XL_2b;

wherein Dg is absent or a divalent moiety of the formula XL₃ ,

XL_2a, or XL_2b ;

or wherein C₈-D₉ is XL₇ , XL_7a, or when X, A₆ and B₇ are absent,

wherein E₁₀-F₁₁ is a divalent moiety of the formula XL₆, XL_6a, XL_6b, XL_6c, XL_6d, XL_6e, XL_6f, or XL_6g;

wherein * indicates an asymmetric center which is either in the R or S configuration; wherein G₁₂ is absent or a divalent moiety of the formula XL₄, XL_4a, or XL_2b;

wherein H₁₃ is absent or a divalent moiety of the formula XL₄ or

XL _2b;

wherein I₁₄ is absent or a divalent moiety of the formula XL₅ or

XL_2b;

wherein Z is

(a) -O-R₁₀,

(b) -N(R₄)R₁₄, or

(c) C₄-C₈cyclic ami_no; wherein R is

(a) isopropyl,

(b) isobutyl, (c) phenylmethyl, or

(d) C₃-C₇cycloalkyl; wherein R₁ is

(a) hydrogen, (b) C₁-C₅alkyl,

(c) aryl,

(d) C₃-C₇cycloalkyl,

(e) -Het,

(f) C₁-C₃alkoxy, or (g) C₁-C₃alkylthio; wherein R₂ is

(a) hydrogen, or

(b) -CH(R₃)R₄; wherein R₃ is (a) hydrogen,

(b) hydroxy,

(c) C₁-C₅alkyl,

(d) C₃-C₇cycloalkyl,

(e) aryl, (f) -Het,

(g) C₁-C₃alkoxy, or (h) C₁-C₃alkylthio; wherein R₄ at each occurrence is the same or different and is (a) hydrogen, (b) C₁-C₅alkyl,

(c) -(CH₂)_p-aryl,

(d) -(CH₂)_p-Het,

(e) -(CH₂)p-C₃-C₇cycloalkyl, or

(f) 1- or 2-adamantyl; wherein R₅ is

(a) C₁-C₆alkyl,

(b) C₃-C₇cycloalkyl,

(c) aryl,

(d) -Het, or (e) 5-oxo-2-pyrrolidInyl; wherein R₆ is

(a) hydrogen,

(b) C₁-C₅alkyl, (c) -(CH₂)_p-aryl,

(d) -(CH₂)_p-Het,

(e) -(CH₂)p-C₃-C₇cycloalkyl, or

(f) 1- or 2-adamantyl; wherein R₇ is

(a) hydrogen,

(b) C₁-C₅alkyl,

(c) hydroxy,

(d) amino C₁-C₄alkyl-, (e) guanidinyl C₁-C₃alkyl-,

(f) aryl,

(g) -Het,

(b) methylthio,

(i) -(CH₂)p-C₃-C₇cycloalkyl, or

(j) amino; wherein R₈ is

(a) hydrogen,

(b) C₁-C₅alkyl,

(c) hydroxy,

(d) aryl,

(e) -Het,

(f) guanidinyl C₁-C₃alkyl-, or

(g) -(CH₂)_p-C₃-C₇cycloalkyl; wherein R₉ is

(a) hydrogen,

(b) hydroxy,

(c) amino C₁-C₄alkyl-, or

(d) guanidinyl C₁-C₃alkyl-; wherein R₁₀ is

(a) hydrogen,

(b) C₁-C₅alkyl,

(c) -(CH₂)_nR₁₆,

(d) -(CH₂)_nR₁₇,

(e) C₃-C₇cycloalkyl,

(f) a pharmaceutically acceptable cation,

(s⁾ -CH(R₂₅)-CH₂-R₁₅, or

(h) -CH₂-CH(R₁₂)-R₁₅; wherein R₁₁ is -R or -R₂; wherein R₁₂ is - (CH₂)_n-R₁₃ ; wherein R₁₃ is

(a) aryl,

(b) amino,

(c) mono-, di or tri-C₁-C₃alkylamino,

(d) -Het,

(e) C₁-C₅alkyl

(f) C₃-C₇cycloalkyl,

(ε) C₂-C₅alkenyl,

(h) C₃-C₇cycloalkenyl,

(i) hydroxy, (j) C₁-C₃alkoxy,

(k) C₁-C₃alkanoyloxy,

(l) mercapto,

(m) C₁-C₃alkylthlo,

(n) -COOH,

(o) -CO-O-C₁-Cgalkyl,

(P) -CO-O-CH₂-(C₁-C₃alkyl)-N(C₁-C₃alkyl)₂,

(q) -CO-NR₂₂R₂₆;

(r) C₄-C₇cyclic amino,

(s) C4-C₇cycloalkylamino,

(t) guanidyl,

(u) cyano ,

(v) N-cyanoguanidyl,

(w) cyanoamino, (x) (hydroxy C₂-C₄alkyl)amino, or

(y) di- (hydroxyC₂-C₄alkyl)amino; wherein R₁₄ is

(a) hydrogen,

(b) C₁-C₁₀alkyl,

(o) -(CH₂)_n-R₁₈,

(d) -(CH₂)_n-R₁₉,

(e) -CH(R₂₅)-CH₂-R_15,

(f) -CH₂-CH(R₁₂)-R₁₅,

(s) (hydroxy C₁-C₈alkyl), or

(h) (C₁-C₃alkoxy) C₁-C₈alkyl; wherein R₁₅ is

(a) hydroxy, (b) C₃-C₇cycloalkyl,

(c) aryl,

(d) amino,

(e) mono-, di- , or tri- C₁-C₃alkylamino, (f) mono- or di- (hydroxy C₂-C₄alkyl)amino,

(g) -Het,

(h) C₁-C₃alkoxy-,

(i) C₁-C₃alkanoyloxy-,

(j ) mercapto, (k) C₁-C₃alkylthio-,

(l) C₁-C₅alkyl,

(m) C₄-C₇cyclic amino,

(n) C₄-C₇cycloalkylamino,

(o) C₁-C₅alkenyloxy, (p) C₃-C₇cycloalkenyl; wherein R₁₆ is

(a) aryl,

(b) amino,

(c) mono- or di- (C₁-C₃alkyl)amino, (d) hydroxy,

(e) C₃-C₇cycloalkyl,

(f) C₄-C₇cyclic amino, or

(g) C₁-C₃alkanoyloxy; wherein R₁₇ is (a) -Het,

(b) C₁-C₅alkenyl,

(c) C₃-C₇cycloalkenyl,

(d) C₁-C₃alkoxy,

(e) mercapto, (f) C₁-C₃alkylthio,

(g) -COOH,

(h) -CO-O-C₁-Cgalkyl,

(i) -CO-O-CH₂-(C₁-C₃alkyl)-N(C₁-C₃alkyl)_2,

(j) -CO-NR₂₂R₂₆, (k) tri-C₁-C₃alkylamino,

(1) guanidyl, (m) cyano ,

(n) N-cyanoguanidyl, (o) (hydroxy C₂-C4alkyl)amino,

(p) di- (hydroxy C₂-C4alkyl)amino, or

(q) cyanoamino; wherein R₁₈ is (a) amino,

(b) mono-, or di- (C₁-C₃alkyl) amino,

(c) C₄-C₇cyclic amino ; or

(d) C₄-C₇cycloalkylamino; wherein R₁₉ is (a) aryl,

(b) -Het,

(c) tri-C₁-C₃alkylamino,

(d) C₃-C₇cycloalkyl,

(e) C₁-C₅alkenyl, (f) C₃-C₇cycloalkenyl,

(g) hydroxy,

(h) C₁-C₃alkoxy,

(i) C₁-C₃alkanoyloxy, (j) mercapto, (k) C₁-C₃alkylthio,

(1) -COOH,

(m) -CO-O-C₁-C₆alkyl,

(n) -CO-O-CH₂-(C₁-C₃alkyl)-N(C₁-C₃alkyl)₂,

(o) -CO-NR₂₂R₂₆, (p) guanidyl,

(q) cyano,

(r) N-cyanoguanidyl,

(s) cyanoamino,

(t) (hydroxy C₂-C4alkyl)amino, (u) di-(hydroxy C₂-C₄alkyl)amino; or

(v) -SO₃H; wherein R₂₀ is

(a) hydrogen,

(b) C₁-C₅alkyl , or (c) aryl-C₁-C₅alkyl; wherein R₂1 is (a) -NH₂, or

(b) -OH; wherein R₂₂ is

(a) hydrogen, or

(b) C₁-C₃alkyl; wherein R₂₃ is (a) -(CH₂)_n-OH,

(b) -(CH₂)_n-NH₂,

(c) aryl, or

(d) C₁-C₃alkyl; wherein R₂₄ is (a) -R₁,

(b) -(CH₂)_n-OH, or (c) -(CH₂)_n-NH₂; wherein R₂₅ is

(a) hydrogen, (b) C₁-C₃alkyl, or

(c) phenyl-C₁-C₃alkyl; wherein R₂₆ is

(a) hydrogen,

(b) C₁-C₃alkyl, or (c) phenyl-C₁-C₃alkyl; wherein R₅₀ is

(a) hydrogen,

(b) C₁-C₅alkyl,

(c) -(CH₂)_p-aryl, (d) -(CH₂)_p-Het,

(e) -(CH₂)_p-C₃-C₇cycloalkyl, or

(f) 1- or 2-adamantyl; wherein R₆₀ and R₆₁ are the same or different and are

(a) hydrogen, (b) C₁-C₅alkyl,

(c) -(CH₂)_p-aryl,

(d) -(CH₂)_p-Het,

(e) -(CH₂)_p-C₃-C₇cycloalkyl, or

(f) 1- or 2-adamantyl; erein R₆₀ and R₆₁ taken together form a spirocycle of the formula

wherein R₇₀ is

(a) -CHR₈₀,

(b) -O-,

(c) -S-,

(d) -SO-,

(e) -SO₂-, or

(f) -NR₈₁; wherein R₈₀ and R₈₁ are the same or different and are

(a) hydrogen,

(b) C₁-C₅alkyl,

. (c) -(CH₂)_p-aryl,

(d) -(CH₂)_p-Het,

(e) -(CH₂)p-C₃-C₇cycloalkyl, or

(f) 1- or 2-adamantyl; wherein R₉₀ is

(a) hydrogen, or

(b) C₁-C₅alkyl; wherein R₁₀₀ is

(a) hydrogen, (b) C₁-C₅alkyl,

(c) aryl,

(d) C₃-C₇cycloalkyl,

(e) -Het,

(f) C₁-C₃alkoxy, or

(s) C₁-C₃alkylthlo ; wherein R₁₁₀ is

(a) hydrogen,

(b) C₁-C₅alkyl, (c) -(CH₂)_p-aryl, or

(d) halogen; wherein R₁₁₁ is

(a) hydrogen,

(b) C₁-C₅alkyl,

(c) -(CH₂)_p-aryl, or

(d) halogen; wherein i is zero to five, inclusive; wherein m is one or two; wherein for each occurrence n is independently an integer of zero to five, inclusive; wherein p is zero to two, inclusive; wherein q is one to five, inclusive; erein r is zero to three, inclusive; erein t is zero to three, inclusive; wherein Q is

(a) -CH₂-,

(b) -CH(OH)-,

(c) -O-, or (d) -S-; and wherein M is

(a) -CO-, or

(b) -CH₂-; wherein aryl is phenyl or naphthyl substituted by zero to three of the following:

(a) C₁-C₃alkyl,

(b) hydroxy,

(c) C₁-C₃alkoxy,

(d) halo, . (e) amino,

(f) mono- or di-C₁-C₃alkylamino,

(g) -CHO, (h) -COOH, (i) COOR₂₆, (j) CONHR₂₆,

(k) nitro, (l) mercapto, (m) C₁-C₃alkylthio, (n) C₁-C₃alkylsulfinyl, (o) C₁-C₃alkylsulfonyl,

(p) -N(R4)-C₁-C₃alkylsulfonyl, (q) SO₃H, (r) SO₂NH₂, (s) -CN, or (t) -CH₂NH₂; wherein -Het is a 5- or 6-membered saturated or unsaturated ring containing from one to three heteroatoms selected from the group consisting of nitrogen, oxygen, and sulfur; and including any bicyclic group in which any of the above heterocyclic rings is fused to a benzene ring, which heterocyclic moiety is substituted with zero to three of the following:

(i) C₁-C₆alkyl, (ii) hydroxy,

(iii) trifluoromethyl, (iv) C₁-C₄alkoxy,

(v) halo , (vi) aryl, (vii) aryl C₁-C₄alkyl-,

(viii) amino,

(ix) mono- or di- (C₁-C₄alkyl)amino, and (x) C₁-C₅alkanoyl; with the overall provisos that: (1) at least one cyclopropyl amino acid of the formula XL_2b or cycloalkyl amino acid non-cleavable transition state insert of the formula XL_2c is present;

(2) R₁₈ or R₁₉ is hydroxy, mercapto, or amino, or a mono-substituted nitrogen containing group bonded through the nitrogen only when n is not one;

(3) R₁₂ is -(CH₂)_n-R₁₃ and n is zero and both R₁₃ and R₁₅ are oxygen-, nitrogen-, or sulfur-containing substituents bonded through the hetero atom, only when the hetero atom is not also bonded to hydrogen; (4) R₁₇ or R₁₉ is -COOH only when n for that moiety is other than zero;

(5) R₁₆ or R₁₇ is an amino-containing substituent, hydroxy, mercapto, or -Het bonded through the hetero atom only when n for that substituent is an integer from two to five, inclusive; (6) when R₁₂ is -(CH₂)_n-R₁₃ and n is zero, then R₁₃ and R₁₅ cannot both be -COOH; and

(7) R₁₇ or R₁₉ is -Het, only when -Het is other than cyclic amino ; or a carboxy-, amino-, or other reactive group-protected form thereof; or a pharmaceutically acceptable acid addition salt thereof.

6. The renin inhibitory peptide of claim 5 wherein X is R₅-0-C(0)-; wherein A₆ is absent; wherein B₇ is absent; wherein C₈ is XL_2b ; wherein D₉ is XL₃ ; wherein E₁₀-F₁₁ is XL₆; wherein G₁₂ is XL_4a; wherein H₁₃ is absent; wherein I₁₄ is absent; wherein Z is -N(R₄)R₁₄; wherein R₁ is C₁-C₅alkyl; wherein R₂ is -CH(R₃)R₄: wherein R₃ is hydrogen; wherein R₄ is (a) hydrogen, or

(b) C₁-C₅alkyl; wherein R₅ is C₁-C₆alkyl; wherein R₇ is -Het; wherein R₁₁ is isopropyl; wherein R₁₄ is -(CH₂)_n-R₁₉; wherein R₁₉ is -Het; wherein R₅₀ is hydrogen; wherein R₆₀ and R₆₁ are the same or different and are (a) hydrogen, or (b) -(CH₂)_p-aryl.

7. N^α- [ (2S ,4S , 5S) -5- [N- [Nα- [ (E) -2-Phenyl-1-(tert-butoxycarbonyl-amino)cyclopropane-1-carbonyl] -L-histidyl] amino] -4-hydroxy-2- isopropyl-7 -methyl-1-oxooctyl]-N-(2-pyridinylmethyl)-L-isoleucinamide, a compound of claim 6.

8. N^α- [ (2S,4S,5S) -5- [N- [N^α-[ (Z) -2-Phenyl-1-(tert-butoxycarbonyl-amino) cyclopropane -1-carbonyl ] - L-histidyl] amino]-4-hydroxy-2-isopropyl-7-methyl-1-oxooctyl]-N-(2-pyridinylmethyl)-L-isoleucinamide, a compound of claim 6.

9. The renin inhibitory peptide of claim 5 having the cycloalkyl non-cleavable transition state insert of the formula XL_2c.

10. [1S-[1α,2β(1R*,2R*)]]3[3-Cyclohexyl-2-[[L-Boc-Phe-L-His(amino)-1-hydroxypropyl]-2,2-dimethylcycloρropanecarboxylic acid, -L- isoleucyl-2-pyrldylmethylamide, a compound of claim 9.

11. The renin inhibitory peptide of claim 5 having the cyclopropyl amino acid of the formula XL_2b and the cycloalkyl amino acid non-cleavable transition state insert of the formula XL_2c.

12. [1S-[1α,2β(1R*,2R*)]]3[3-Cyclohexyl-2-[[(Z)-(2-phenyl-1-(boc)a-minocyclopropane-1-carboxyl)-L-His (amino)-1-hydroxypropyl]-2,2-dimethylcyclopropanecarboxyllc acid, -L-isoleucyl-2-pyrldylmethylamide, a compound of claim 11.