AU711035B2 - Difluorostatone antiviral agents - Google Patents
Difluorostatone antiviral agents Download PDFInfo
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- AU711035B2 AU711035B2 AU28189/95A AU2818995A AU711035B2 AU 711035 B2 AU711035 B2 AU 711035B2 AU 28189/95 A AU28189/95 A AU 28189/95A AU 2818995 A AU2818995 A AU 2818995A AU 711035 B2 AU711035 B2 AU 711035B2
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- methyl
- difluoro
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- C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
- C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
- C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
- C07D213/24—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with substituted hydrocarbon radicals attached to ring carbon atoms
- C07D213/28—Radicals substituted by singly-bound oxygen or sulphur atoms
- C07D213/30—Oxygen atoms
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
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- C07C237/00—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by amino groups
- C07C237/02—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by amino groups having the carbon atoms of the carboxamide groups bound to acyclic carbon atoms of the carbon skeleton
- C07C237/22—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by amino groups having the carbon atoms of the carboxamide groups bound to acyclic carbon atoms of the carbon skeleton having nitrogen atoms of amino groups bound to the carbon skeleton of the acid part, further acylated
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- C07C271/06—Esters of carbamic acids
- C07C271/08—Esters of carbamic acids having oxygen atoms of carbamate groups bound to acyclic carbon atoms
- C07C271/10—Esters of carbamic acids having oxygen atoms of carbamate groups bound to acyclic carbon atoms with the nitrogen atoms of the carbamate groups bound to hydrogen atoms or to acyclic carbon atoms
- C07C271/22—Esters of carbamic acids having oxygen atoms of carbamate groups bound to acyclic carbon atoms with the nitrogen atoms of the carbamate groups bound to hydrogen atoms or to acyclic carbon atoms to carbon atoms of hydrocarbon radicals substituted by carboxyl groups
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- C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
- C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
- C07D213/24—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with substituted hydrocarbon radicals attached to ring carbon atoms
- C07D213/36—Radicals substituted by singly-bound nitrogen atoms
- C07D213/40—Acylated substituent nitrogen atom
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- C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
- C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
- C07D213/60—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
- C07D213/78—Carbon atoms having three bonds to hetero atoms, with at the most one bond to halogen, e.g. ester or nitrile radicals
- C07D213/81—Amides; Imides
- C07D213/82—Amides; Imides in position 3
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- C07D231/02—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings not condensed with other rings
- C07D231/10—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members
- C07D231/12—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached to ring carbon atoms
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- C07D233/54—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members
- C07D233/56—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members with only hydrogen atoms or radicals containing only hydrogen and carbon atoms, attached to ring carbon atoms
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- C07D249/02—Heterocyclic compounds containing five-membered rings having three nitrogen atoms as the only ring hetero atoms not condensed with other rings
- C07D249/08—1,2,4-Triazoles; Hydrogenated 1,2,4-triazoles
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- C07D295/00—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms
- C07D295/04—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms
- C07D295/14—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
- C07D295/145—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals with the ring nitrogen atoms and the carbon atoms with three bonds to hetero atoms attached to the same carbon chain, which is not interrupted by carbocyclic rings
- C07D295/15—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals with the ring nitrogen atoms and the carbon atoms with three bonds to hetero atoms attached to the same carbon chain, which is not interrupted by carbocyclic rings to an acyclic saturated chain
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- C07D295/16—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms acylated on ring nitrogen atoms
- C07D295/18—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms acylated on ring nitrogen atoms by radicals derived from carboxylic acids, or sulfur or nitrogen analogues thereof
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- C07D295/18—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms acylated on ring nitrogen atoms by radicals derived from carboxylic acids, or sulfur or nitrogen analogues thereof
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- C07D295/22—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with hetero atoms directly attached to ring nitrogen atoms
- C07D295/26—Sulfur atoms
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Description
WO 96/02499 PCT/US95/07241 -1- DIFLUOROSTATONE ANTIVIRAL AGENTS BACKGROUND OF THE INVENTION Retroviruses are a class of viruses which transport their genetic material as ribonucleic acid rather than as deoxyribonucleic acid. Retroviruses are associated with a wide variety of diseases in man, one of which is AIDS.
Although there have been disclosures of other anti-viral agents useful in the treatment of AIDS, for example see patent applications EP 0 218 688, EP 0 352 000 and PCT/US 91/09741, the compounds of the present invention have not been previously disclosed. PCT/US 91/09741 is hereby incorporated by reference.
SUMMARY OF THE INVENTION The present invention relates to compounds having the following general formula P2 P1 I I R1--CNH-CH--CNH-CH-C-CF 2 -C-NR5R6 II II II II O O O O 0 0X formula I and the stereoisomers, hydrates, isosteres and the pharmaceutically acceptable salts thereof wherein WO 96/02499 PCT/US95/07241 -2- Pi is
T
C
1 6 alkylenewherein T is and T' is or hydrogen, wherein each of W and W' are independently
C
1 6 alkylene or nothing, provided that W is C2-6 alkylene when W is directly attached to a nitrogen atom in R, provided that W' is C2- 6 alkylene when W' is directly attached to a nitrogen atom in R', provided that W or W' are each independently C 1 6 alkylene when R or R' are each independently an aryl; P2 is C 1 -6 alkyl, cyclopentyl, hydroxy C1_ 6 alkyl, phenyl, benzyl or 3-tetrahydrofuryl; R and R' are each independently -CH 2 CHO, hydroxy CI- 6 alkyl, C 1 6 alkoxy Ci- 6 alkyl, C 1 -6 alkyl, (R3)d
C
1 6 alkenylene, piperazinyl, substituted piperazinyl, piperidyl, morpholinyl, pyridyl, pyrazinyl, pyrimidinyl or phenyl wherein substituted piperazinyl is piperazinyl substituted on one nitrogen atom thereof with CHO, C(O)NHR' 4 C1- 4 alkyl or C0 2
R
4
R
1 is N NN
C
2 O N H 2 )O 0 Ul- (CH 2
)O-
0 0 N
N
0 N 0 0 0 N N C0 2
R'
4
OHN
(CH
2 2 -CH- -CH 2
C=H
O. *CH OH i0 Cl6aklpeyo ez R' is hyrgno l6akl 3A
R
5 is hydrogen, Cl..
1 5 alkyl, OH, hydroxy Cl1 alkyl, 01.6 alkoxy, -CH([(CH 2 )d-O-CH 2 x-
R'
8 2
-CH
2 Si(CH 3 2
(R
3 PDL, -(C 1 6 alkylene)-0R 4 tVOD\ALDEEEOTES2189.O WO 96/02499 WO 9602499PCTIUS95/07241 j i uroe 4. r
I
-4alkyly OH, hydromy C±1-15 a4-k-y1, RI8) 2
-GH
2 Si (GH 3 2 (R3) I -PD4)' Me ~HO 1 (CH2)b
N.
(b) t 6 %T 4
C
1 6 alkylene- (a) (c) or CH2-K N N (d) wherein PDL is 3- or 4-pyridyl, or psubstituted benzyloxy, wherein the substitution is with a nitro, OH, amino, C 1 6 alkoxy, hydroxy CI- 6 alkylene, or halogen; Y is Cl- 15 alkyl, hydroxy Cl..
15 alkyl, C1-.
6 alkyl or -(CH2)e-C6H4-(V)e'; Z is -(CH 2 )d-O-CHO, C 1 6 alkyiene-O-(CH 2 )d-(O-CH2-CH2)e-O-C1-6 alkyl, CHO, C0 2
R
4
CO
2
NHR
4 -(CH2)d0I-CH2)d'-R'7, -(CH2)e-OR4 or (CH2)e wherein V is OR 4 or hydroxy C 1 6 alkylene; provided that d'=2 when R'7 is piperazinyl, substituted piperazinyl, piperidyl or morpholinyl; WO 96/02499 PCTUS95/07241
R
6 is as defined for R 5 with the proviso that R 6 is other than hydrogen when R 5 is hydrogen, or R 5 and R 6 are taken together with the nitrogen atom to which they are attached are selected from the group consisting of; -N -N 0 (h) (CH2)b R
R
3 -Si
H
/N 3 I (j (k) R7
R
-N \-CHO -N or -N R 8 (n)
R
7 is CH20R 4
C(O)NHR
4 or CHO; R'7 is piperazinyl, substituted piperazinyl, piperidyl, morpholinyl, pyridyl, pyrazinyl, pyrimidinyl or phenyl, wherein substituted piperazinyl is piperazinyl substituted on one nitrogen atom thereof with CHO,
C(O)NHR
4 C1- 4 alkyl or C0 2
R
4
R
8 is OH) or =0;
R'
8 is pyrimidyl, pyridyl, pyrazinyl or phenyl; WO 96/02499 PCT/US95/07241 -6a is zero, 1, 2 or 3; b and b' are each independently zero or 1; d and d' are each independently 1 or 2; e and e' are each independently zero, 1 or 2; and x is zero or one.
DETAILED DESCRIPTION OF THE INVENTION The terms "halo", "halogen" or "halide" refer to a chlorine, bromine or iodine atom.
Isosteres of the compounds of Formula I include those wherein the a-amino acid residues of the P 1 and P 2 substituents are in their unnatural configuration (when there is a natural configuration) or when the normal peptide amide linkage is modified, such as for example, to form 0
II
-CH
2 NH- (reduced), -C-N(CH 3 (N-methylamide), -COCH 2 (keto), -CH(OH)CH 2 (hydroxy), -CH(NH 2
)CH
2 (amino),
-CH
2
CH
2 (hydrocarbon). Preferably a compound of the invention should not be in an isosteric form. Unless otherwise stated the a-amino acids are preferably in their L-configuration.
A compound of the invention may be in free form, e.g., amphoteric form, or in salt, acid addition or anionic salt, form. A compound in free form may be converted into a salt form in an art-known manner and vice-versa.
The pharmaceutically acceptable salts of the peptide of Formula I (in the form of water, or oil-soluble or dispersible products) include the conventional non-toxic salts or the quaternary ammonium salts of these peptides, which are formed, from inorganic or.organic acids or bases. Examples of such acid addition salts include WO 96/02499 PCT/US95/07241 -7acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, citrate, camphorate, camphorsulfonate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, fumarate, glucoheptanoate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, oxalate, paemoate, pectinate, persulfate, 3-phenylpropionate, picrate, pivalate, propionate, succinate, tartrate, thiocyanate, tosylate, and undecanoate. Base salts include ammonium salts, alkalimetal salts such as sodium and potassium salts, alkaline earth metal salts such as calcium and magnesium salts, salts with organic bases such as dicyclohexylamine salts, N-methyl- D-glucamine, and salts with amino acids such as arginine, lysine, and so forth. Also, the basic nitrogen-containing groups may be quaternized with such agents as lower alkyl halides, such as methyl, ethyl, propyl, and butyl chloride, bromides and iodides; dialkyl sulfates like dimethyl, diethyl, dibutyl; and diamyl sulfates, long chain halides such as decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides, aralkyl halides like benzyl and phenethyl bromides and others.
The hydrates of the compounds of Formula I are hydrated compounds having the partial structure t I 1 .CF2\ ^j CF O SHO OH o and in their end-use application are generally the active Sforms.
WO 96/024199 PCTJUS9507241 -8- In general, as used herein, the term "alkyl" includes the straight, branched-chain and cyclized manifestations thereof unless otherwise indicated, particularly such moieties as methyl, ethyl, isopropyl, n-butyl, t-butyl, -C51 2 -t-butyl, cyclopropyl, n-propyl, pentyl, cyclopentyl, n-hexyl, cyclohexyl and cyclohexylmethyl. The term aralkyl", when used, includes those aryl moieties attached to an alkylene bridging moiety, preferably methyl or ethyl.
"Aryl" includes both carbocyclic and hetereocyclic moieties of which phenyl, pyridyl, pyrimidinyl, pyrazinyl, indolyl, indazolyl, furyl and thienyl are of primary interest; these moieties being inclusive of their position isomers such as, for example, or 4-pyridyl, 2- or 3-furyl and thienyl, or 3-indolyl or the 1- and 3indazolyl, as well as the dihydro and tetrahydro analogs of the furyl and thienyl moieties. Also included within the term "aryl" are such fused carbocyclic moieties as pentalenyl, indenyl, naphthalenyl, azulenyl, heptalenyl, acenaphthylenyl, fluorenyl, phenalenyl, phenanthrenyl, anthracenyl, acephenanthrylenyl, aceanthrylenyl, triphenylenyl, pyrenyl, chrysenyl and naphthacenyl. Also included within the term "aryl" are such other heterocyclic radicals as 2- or 3-benzo[b]thienyl, 2- or 3-naphtho[2,3blthienyl, 2- or 3-thianthrenyl, 2H-pyran-3-(or 4- or l-isobenzo- furanyl, 2H-chromenyl-3-yl, 2- or 3phenoxathiinyl, 2- or 3-pyrrolyl, 4- or 3-pyrazolyl, 2-pyrazinyl, 2-pyrimidinyl, 3-pyridazinyl, 2-indolizinyl, 1-isoindolyl, 4H-quinolizin-2-yl, 3-isoquinolyl, 2quinolyl, l-phthalazinyl, l,8-naphthyridinyl, 2quinoxalinyl, 2-quinazolinyl, 3-cinnolinyl, 2-pteridinyl, 4aH-carbazol-2-yl, 2-carbazolyl, B-carbolin-3-yl, 3-phenanthridinyl, 2-acridinyl, 2-perimidinyl, 1-phenazinyl, 3-isothiazolyl, 2-phenothiazinyl, 3 -isoxazolyl, 2-phenoxazinyl, 3-isochromanyl, 7-chromanyl, 2-pyrrolin-3-yl, 2-imidazolidinyl, 2-imidazolin-4-yl, 2 -pyrazolidinyl, 3 -pyrazolin-3-yl, 2-piperidyl, WO 96/02499 PCT/US95/07241 -9- 2-piperazinyl, 1-indolinyl, 1-isoindolinyl, 3-morpholinyl, benzo[b]isoquinolinyl and benzo[b]furanyl, including the position isomers thereof.
Likewise the term "alkylene" includes straight or branched-chain moieties. Some examples of branched-chain alkylene moieties are ethylethylene, 2-methyltrimethylene, 2,2-dimethyltrimethylene, and so on. For example, C 3 alkylene can mean CH3
-CH
2
-CH
2
-CH
2 or or -CH 2 -CH- or -CH-CH 2 I I I
CH
3 CH 3
CH
3 All (Cg 6 moieties such as C 1 -6 alkyl, C1_ 6 allenyl, C1-6 alkoxy, and hydroxy C1_ 6 alkyl, are more preferably C 1 3 moieties (containing 1-3 carbon atoms instead of 1-6 carbon atoms).
The fluorenylmethyloxy moiety is that moiety generally called by its abbreviation FMOC, and is the fluorenyl moiety bearing -CH20 attached to the 9-position of the fluorenyl moiety. Other terms defined herein are piperazinyl -N NH) or substituted piperazinyl -N N-*r the substitution occurring only at one nitrogen atom which is not attached to the remainder of the molecule (attachment via a nitrogen atom). The substituents are one of CHO, C(O)NHR 4
C
1 4 alkyl or C0 2
R
4 The term "Bn" refers to a benzyl functionality of the formula;
CH
2 WO 96/02499 PCT/US95/07241 Piperidinyl and morpholinyl both bind to the rest of -N -N O molecule via their respective nitrogen atoms while pyrimidinyl, pyridyl and pyrazinyl bind to the rest N Np
N
of the molecule anywhere except their respective nitrogen atoms.
More specifically, in the instance wherein P 2 is either
C
1 i_ alkyl or hydroxy C1_ 6 alkyl, such moieties as -C(CH 3 3
-CH(CH
3 2
-CH(CH
3
)(C
2
H
5
-C(OH)(CH
3 2 and -CH(OH)CH 3 are preferred. The "hydroxy C 1 6 alkyl" moiety is illustrated in one example by -CH 2 -OH, the "C 1 -6 alkoxy C 1 6 alkyl" moiety, is illustrated in one example by -CH 2
-OCH
3 (although in each instance the C 1 -6 alkylene may be straight or branched and the hydroxy radical is not limited to the terminal carbon atom of the alkyl moiety).
As it is often quite advantageous to have what is termed an amino protecting group the scope of those compounds of Formula I includes those R 1 moieties which, together with their adjacent carbonyl moiety form such groups as acetyl succinyl (Suc), benzoyl (Bz), t-butyloxycarbonyl (Boc), benzyloxycarbonyl (CBZ), tosyl dansyl (DNS), isovaleryl (Iva), methoxysuccinyl (MeOSuc), l-adamantanesulphonyl (AdSO 2 1-adamantaneacetyl (AdAc), phenylacetyl, t-butylacetyl (Tba), bis[(lnaphthyl)methyl]acetyl (BNMA) and Rz wherein Rz is an aryl group as previously described suitably substituted by 1 to 3 members selected independently from the group consisting of fluoro, chloro, bromo, iodo, trifluoromethyl, hydroxy, alkyl containing from 1 to 6 carbons, alkoxy containing WO 96/02499 PCT/US95/07241 -11from 1 to 6 carbons, carboxy, alkylcarbonylamino wherein the alkyl group contains 1 to 6 carbons, 5-tetrazolo, and acylsulfonamido acylaminosulfonyl and sulfonylaminocarbonyl) containing from 1 to 15 carbons, provided that when the acylsulfonamido contains an aryl, the aryl may be further substituted by a member selected from fluoro, chloro, bromo, iodo and nitro.
Among the classes of amino protecting groups contemplated are: acyl type protecting groups such as formyl, trifluoroacetyl, phthalyl, p-toluenesulfonyl (tosyl), benzenesulfonyl, nitrophenylsulfenyl, tritylsulfenyl, O-nitrophenoxyacetyl, and a-chlorobutyryl; aromatic urethane type protecting groups such as benzyloxycarbonyl and substituted benzyloxycarbonyls such as p-chlorobenzyloxycarbonyl, p-methoxybenzyloxycarbonyl, p-nitrobenzyloxycarbonyl, p-bromobenzyloxycarbonyl, l-(p-biphenylyl)-l-methylethoxycarbonyl, a-dimethyland benzhydryloxycarbonyl; aliphatic urethane protecting groups such as tertbutyloxycarbonyl (Boc), diisopropylmethoxycarbonyl, isopropyloxycarbonyl, ethoxycarbonyl, and allyloxycarbonyl; cycloalkyl urethane type protecting groups such as cyclopentyloxycarbonyl, adamantyloxycarbonyl, and cyclohexyloxycarbonyl; thio urethane type protecting groups such as phenylthiocarbonyl; alkyl type protecting groups such as triphenylmethyl (trityl) and benzyl (Bn); trialkylsilane protecting groups such as trimethylsilane if compatible. The preferred a-amino protecting groups are tert-butyloxycarbonyl (Boc) or benzyloxycarbonyl (CBZ). The use of Boc as an a-amino protecting group for amino acids is described by Bodansky et al. in "The Practice of Peptide Synthesis", Springer-Verlag, Berlin (1984), p. The term "stereoisomers" is a general term for all isomers of individuals molecules that differ only in the orientation of their atoms in space. It includes mirror 12 image isomers (enantiomers), geometric (cis/trans) isomers, and isomers of compounds with more than one chiral centre that are not mirror images of one another (diastereoisomers). For amino-acids, the designations L/D, or R/S can be used as described in IUPAC-IUB Joint Commission on Biochemical Nomenclature, Eur. J.
Bicochem. 138:9-37 (1984).
Throughout the description and claims of this specification the word "comprise" and variations of the word, such as "comprising" and "comprises" is not intended to exclude other additives, components, integers or steps.
In general the compounds of this invention may be prepared using standard chemical reactions analogously known in the art. More specifically, the preparation of compounds of structure is well known in the art and described generally by Schirlin, D. and Van Dorsselaer, V. in PCT/US91/09741 published July 23, 1992 with an international publication number of WO 92/12123.
The compounds of formula can be prepared as described in Reaction Schemes A B, C and D. All the substituents, unless otherwise indicated, are previously defined. The reagents and starting materials are readily available to one of ordinary skill in the art.
e *a 'c y W ORD\MARLO\NODELETE\OTHERS\2818995.DOC REACTION SCHEME A
PI
PgNHCH-CHO 1
P
1 PgNHCH-jHCF 2 fjjOEt OH 0 (ci)
POH
PgNHCH- H-CF 2 -jj-Ot OH 0 (3) (4) I1(b) ~1(b)
P
1 PgqNH-CH-CH-CF 2 -CNR 5
R
6 OH 0 PgNH-CH-CH-C2 -RR OH 0 Poll PgNH-C H-CH-CF CN5R6 OH 0 (14) 1 (d) (6) (d)4 (5) Cc 2 1
P
1
H
2
-CH--CH-CF
2 -C-NR 5
R
6 OH 0 (7)
P
1
NH
2
-CH-CH-CF
2
-C-NR
5
R
6 OH 0 (9)
POH
N -C-HCF 2 -C-NR 5 R 6 OH 0 REACTION SCHEME A' (7) (9) I (e) I (e) I (e) P2 P 2 2 I I I I I I Re 1 C-NH-CH-C-NH-CH-CH-CF 2 -CNR 5 R 6 Re 1
C-NH-CH-C-HH-CH-CH-CF
2
-CNR
5 R 6 R-iC-NH-C;H-C-nH-UH-t-C 0 O 0 0 0 OH 0 00 OH 0 (10) (1IKILu (f)$I (C2) 2 p 1 R ii n 2
-CNR
5
R
6
IA
(9) R# N-H-LCH-j NHCHCCF 2
NR
5
R
6 00 0 0
IA'
REACTION SCHEME A" (7) (9) I (e) I (e) 1 (e) p 1
III
0 OH 0 (11)) (f) 4(C) p 1 R 1 C-NH-CHi-CH-CF 2
-C-NR
5
R
6 0 OH 0 R' IC-NH-CH-CH-CF 2
-C-NR
5
R
6 11 1 11 0 OH 0 .0(7) (12) (C2)
P
1 R C -N -L~u-C Fr 2 -CNR 5
R
6 (g) 1 11H1-CH p 1
C
2 CjNR 5
R
6 0 0 0
IB
WO 96/02499 PCT/US95/07241 -16- In Scheme A, step the aldehyde of formula is subjected to a condensation reaction under Reformatski conditions with an ester of bromodifluoroacetic acid, preferably the ethyl ester in the presence of zinc and in an anhydrous aprotic solvent, tetrahydrofuran, ether, dimethoxyethane and the like under a nitrogen or argon inert atmosphere. The reaction is gently heated to about for about 1-12 hours or ultrasonicated to produce compounds Alternatively, in Reaction Scheme A step the condensation to produce compounds can be achieved in greater yields and at lower reaction temperatures utilizing the following general method. Under an inert atmosphere, such as nitrogen, the aldehyde is dissolved in a suitable anhydrous organic solvent. Examples of a suitable anhydrous organic solvent are tetrahydrofuran, diethyl ether, t-butyl methyl ether and the like. The solution is cooled to approximately 0°C. To the solution is added about 0.30 equivalents of silver acetate, about 2.1 equivalents of zinc dust, and about 2 equivalents of ethyl bromodifluoroacetate. About 0.34 equivalents of diethylaluminum chloride (as a solution in toluene) is added slowly to the reaction keeping the temperature of the reaction below 12 0 C. The reaction is allowed to stir for 1 to 3 hours at about 0°C and then at room temperature for 4 to 12 hours. The reaction is then cooled to about 10 0 C and quenched with saturated aqueous ammonium chloride. The compound is then isolated and purified by techniques well known in the art. For example a solution of sodium hydrogen tartrate is added and the reaction is allowed to warm from 10 0 C to room temperature. The mixture is filtered, the solids washed with a suitable organic solvent, such as ethyl acetate and the layers of the filtrate are separated. The aqueous layer is extracted with ethyl acetate, the organic layer and extracts are WO 96/02499 PCT/US95/07241 -17combined, dried over anhydrous magnesium sulfate, filtered and concentrated. The residue is purified by flash chromatography on silica gel with a suitable eluent, such as cyclohexane/ethyl acetate to provide the compounds In Scheme A, step formation of compounds or (14) may be effected directly or undirectly. In one method, the esters of formula or (13) are de-esterified using a suitable base, such as LiOH, KOH, NaOH and the like, in the presence of water and a partially water miscible solvent (such as tetrahydrofuran, dimethoxyethane, dioxane) at about room temperature. The resulting acid can then be aminated with the appropriate R 5
R
6 -substituted amine using standard peptide-like coupling conditions. The selection of the appropriate coupling reaction procedure is within the skill of the art. The coupling reaction can be carried out using standard coupling procedures such as the azide method, mixed carbonic acid anhydride (isobutyl chloroformate) method, carbodiimide [dicyclohexylcarbodiimide, diisopropylcarbodiimide, or water-soluble carbodiimide,l-(3-dimethylaminopropyl)-3ethylcarbodiimide hydrochloride (EDC)] method, active ester (p-nitrophenyl ester, N-hydroxy-succinic imido ester) method, Woodward reagent K method, carbonyldiimidazole method, phosphorus reagents such as BOP-C1, or oxidationreduction methods. Some of these methods (especially the carbodiimide method) can be enhanced by adding hydroxybenzotriazole (HOBT), for example the mixed anhydride method may be employed, using DCC and hydroxybenzotriazole at room temperature in solvents such as CH 2 C1 2 tetrahydrofuran or dimethylformamide.
Alternatively the esters or (13) may be directly subjected to a reaction with the appropriate R 5
R
6 substituted amine without or with a solvent (tetrahydrofuran) at a temperature of from 0 to 80 0
C.
WO 96/02499 PCT/US95/07241 -18- Alternatively, an appropriate R 5
,R
6 -substituted amine that is protected as necessary is dissolved in a suitable organic solvent, such as dichloromethane under an inert atmosphere, such as nitrogen. An equivalent of a 2M solution of trimethylaluminum in toluene is added dropwise to the solution. After approximately 15 minutes this solution is added to approximately 0.3 equivalents of ester or (13) dissolved in a suitable organic solvent, such as dichloromethane. The reaction is allowed to stir for about 15 to 24 hours at about room temperature to 40 0 C. The product is then isolated using techniques well known in the art. For example cold dilute aqueous hydrochloric acid and ethyl acetate is added. The organic layer is separated and washed with water, brine, dried over anhydrous magnesium sulfate, filtered and concentrated under vacuum to provide the compounds or (14).
In Step compounds or (11) are prepared by removal of the P' 1 protecting group using standard procedures well known in the art [see T.H. Green, "Protective Groups in Organic Synthesis", John Wiley and Sons, 1981], such as hydrogenation. The free phenol functionality is then reacted with an appropriate alkyl halide in an inert solvent (preferably anhydrous dioxane, anhydrous acetone or anhydrous dimethylformamide) in the presence of a base (potassium or cesium carbonate) with or without potassium iodide at room or reflux temperature.
In Step (ci) compound (13) is prepared by removal of the P' 1 protecting group using standard procedures well known in the art[see T.H. Green, "Protective Groups in Organic Synthesis", John Wiley and Sons, 1981], such as, hydrogenation, PoH being the compound obtained. POH being a free phenol.
In Step (c2) compounds or (11) are prepared from the PoHderivatives (16) or (17) by reaction with WO 96/02499 PCTIUS95/07241 -19an appropriate alkylhalide in an inert solvent, in the presence of a base. For example, the PoHderivative is dissolved in a suitable organic solvent, such as acetone.
Approximately 1.2 equivalents of a suitable base, such as potassium carbonate, are added followed by addition of approximately 1.15 equivalents of the alkyl halide. A catalytic amount of potassium iodide is then added and the reaction is stirred for 1 to 3 days. The product is isolated and purified by techniques well known in the art, such as extractive methods and recrystallization. For example, the reaction is poured into a suitable solvent mixture, such as ethyl acetate/dilute aqueous sodium chloride and the organic layer is separated. The organic layer is then washed with dilute aqueous potassium hydroxide, brine, dried over anhydrous magnesium sulfate, filtered and concentrated under vacuum. The residue is purified by recrystallization from a suitable solvent mixture, such as cyclohexane/ethyl acetate to provide compounds or (11).
In Step for the preparation of Compounds (9) and the protecting groups Pg may readily be removed by standard procedures well known in the art [see T.H.
Green, "Protective Groups in Organic Synthesis", John Wiley and Sons, 1981], preferably acid/base hydrolysis formic acid at room temperature followed by extraction of the free base after treatment with sodium carbonate).
In Step Compounds or (15) are subjected to a peptide coupling procedure with an appropriately protected acid of the formula R' 1
CONHCH(P
2
)CO
2 H or R' 1 C0 2
H,
using the herein-described procedures (or by any other coupling procedure well known in the art, or as described in European Patent Application, Serial Number 93 401 785.6) to produce compounds and (11) (from compound and (12) (from compound and (16) and (17) (from compound R' 1 is defined as R 1 except for those WO 96/02499 PCT/US95/07241 instances wherein protection of the R 1 group may be required as recognized by one of ordinary skill in the art. For example, a hydroxyl functionality on the alkyl portion of the R 1 group must be protected prior to the oxidation in step In Step the oxidation of compounds and (11) may be effected by methods well known in the art, such as the Swern oxidation procedure, or with l,l,l-triacetoxy-l,1dihydro-l,2-benziodoxol-3(lH)-one to provide compounds of formulas IA and IB.
In general the Swern oxidation [see Synthesis, (1981), 165] is effected by reacting about 2 to 20 equivalents of dimethylsulfoxide (DMSO) with about 1 to 10 equivalents of trifluoroacetic anhydride [(CF 3
CO)
2 0] or oxalyl chloride [(COC1) 2 said reactants being dissolved in an inert solvent, methylene chloride (CH 2 C12), said reaction being under an inert atmosphere nitrogen or equivalently functioning gas) under anhydrous conditions at temperatures of about -70 0 C to -30 0 C to form an insitu sulfonium adduct to which is added about 1 equivalent of the appropriate alcohols, compounds and (11).
Preferably, the alcohols are dissolved in an inert solvent,
CH
2 C1 2 tetrahydrofuran, or minimum amounts of DMSO, and the reaction mixture is allowed to warm to about -50 0
C
or -20 0 C (for about 20-60 minutes) and then the reaction is completed by adding about 3 to 30 equivalents of a tertiary amine, triethylamine, diisopropylethylamine, N-methyl morpholine, etc.
Alternatively the oxidation can be carried out with the Dess-Martin periodinane l,l,l-triacetoxy-l,ldihydro-l,2-benziodoxol-3(1H)-one), [see Dess Martin, J.
Org. Chem., 48, 4155, (1983)]. This oxidation is effected by contacting about 1 equivalent of the alcohol with 1 to equivalents of periodinane (preferably greater than WO 96/02499 PCT/US95/07241 -21equivalents), said reagent being in suspension in an inert solvent methylene chloride) under an inert atmosphere (preferably nitrogen) under anhydrous conditions at 0°C to 50 0 C (preferably room temperature) and allowing the reactants to interact for about 1 to 48 hours.
Optional deprotection of the amine protecting groups may be effected as desired after the ketones have been isolated.
In general, the modified Jones oxidation procedure may conveniently be effected by reacting the alcohols with pyridinium dichromate by contacting the reactants together in a water-trapping molecular sieve powder, a grounded 3 Angstr6m molecular sieve), wherein said contact is in the presence of glacial acetic acid at about 0°C to 50 0 C, preferably at room temperature followed by isolation and then optionally removing amine protecting groups.
Alternatively, 1 to 5 equivalents of a chromic anhydride-pyridine complex a Sarett reagent prepared insitu) [see Fieser and Fieser "Reagents for Organic Synthesis" Vol. 1, pp. 145 and Sarett, et al., J.A.C.S. 422, (1953)] in an inert solvent CH 2 C12) under an inert atmosphere under anhydrous conditions at 0 C to 50 0
C
is treated with 1 equivalent of the alcohol. The reaction is allowed to stir for about 1 to 15 hours. The product is isolated and purified by techniques well known in the art followed by optionally removing amine protecting groups.
In step the compounds decribed by formulas IA and IB wherein R' 1 is protected as required, are deprotected under conditions well known in the art [see T.H. Green, "Protective Groups in Organic Synthesis", John Wiley and Sons, 1981], to provide compounds of formula IA' and IB' wherein R"I are those substituents which required protection as required. For example the protected compound is dissolved in a suitable organic solvent, such as methylene chloride and treated with TFA/H 2 0. The reaction WO 96/02499 PCT/US95/07241 -22is allowed to stir at room temperature for about 4 to hours and then it is concentrated under vacuum. The residue is purified by techniques well known in the art, such as extractive methods followed by flash chromatography (silica gel, hexane/ethyl acetate) to provide the deprotected compound.
For the preparation of the necessary aldehydes of formula and the acids which are to be coupled with the compounds or alternative alkylation procedures are utilized depending upon whether the P 1 and/or the P 2 moieties are or are not residues of natural amino acids. For the preparation of these intermediates wherein the P 1 or P 2 moieties are residues of natural amino acids (or minor modifications thereof, P 1 or P 2 being a benzyl or methyl ether of tyrosine), the compounds are either known or are prepared by processes and techniques well known in the art.
To prepare the intermediates of the formula PgHN-CHCO 2
R
9 wherein Pg is an amino protecting group, P 3 is either a P'1 or P' 2 moiety with P' 1 and P' 2 being as defined for P 1 and
P
2 respectively, except that they are other than residues of naturally occuring amino acids, and the R 9 moiety is an alkyl radical, preferably methyl when P 3 is P' 1 and ethyl when P 3 is P' 2 alternative methods are available.
To prepare the intermediates of formula WO 96/02499 PCT/US95/07241 -23- ,1 P 1 2 PgHN-CHCHO PgHN-CHCOOH the following reaction scheme may be utilized REACTION SCHEME B P3 0 PgNHCH 2
CO
2
R
9 Base PgNHCHCO 2
R
9 (18) P 3 X (19) 1 wherein P 3 is as previously defined and X is a leaving group, preferably halo or triflate, R 9 is methyl when P 3 is
P'
1 and ethyl when P 3 is P' 2 In essence, the preparation of compounds (19) utilizes the Krapcho method [Tetrahedron Letters, 26, 2205 (1976)] for alkylation wherein compounds (18) are treated with a base, LDA, (lithium diisopropylamide), followed by reaction with the desired P 3 X in the presence of TMEDA tetramethylethylenediamine) in a solvent (tetrahydro- 2 furan) with or without HMPA hepamethylphosphonamide) according to the standard Krapcho conditions. Following alkylation the compounds are then subjected to a reduction using diisobutyl alaminum hydride (Dibal) in a mixture of solvents, ether, toluene, hexane, tetrahydrofuran at 3 about -78 0 C for about 1 hour. Following the preparation of the aldehydes of Formula (10B), the compounds are subjected to the processes of Reaction Schemes A, A' and/or A".
Alternatively, the compounds of (19) may be prepared by a Malonate/Curtius type sequence of reactions, [see Yamada, et al., J. Amer. Chem. Soc., (1972) 94, 6203] as illustrated by the following reaction scheme 1 WO 96/02499 PCT/US95/07241 -24- REACTION SCHEME C t-BuO 2
CCH
2
CO
2
R
9 Base t-BuO 2 CCH2CO 2
R
9 5(20)
P
3 X (21) SRemoval of t-Bu
P
3
HO
2
CCH
2
CO
2
R
9 (22) Curtius-type rearrangement (19) wherein t-Bu is t-butyl, although other selectively removable acid protecting groups may be utilized, and P 3
X
is as previously defined. This reaction involves the alkylation of the malonate ester (20) followed by selective removal of the t-butyl protecting group to produce compounds These compounds are then transformed to (19) using the Curtius type rearrangement which entails their conversion to the protected amine via the intermediately formed acylazides and isocyanates. The resultant amines are then protected with standard amino protecting groups, preferentially being protected insitu.
In the instance wherein P 3 represents a P' 1 moiety, the ester is transformed to the desired aldehydes of Formula using standard Dibal reduction techniques, particularly in this situation (wherein P 1 is not a residue of a natural amino acid). Alternatively, (as is preferred when P 1 is a residue of a natural amino acid) the ester is de-esterified to its corresponding acid, converted to its corresponding hydroxamate and the hydroxamate upon treatment with lithium aluminum hydride is converted to its aldehyde. In the instance wherein P 3 represents a P' 2 WO 96/02499 PCTUS95/07241 moiety, the ethyl ester of compounds (19) are removed and the resulting compounds are ready for coupling as outlined in Reaction Scheme A'.
In Scheme D an alternative procedure for preparation of compounds of formula IA is set forth.
WO 96/02499 PCT/US95/07241 -26- Reaction Scheme D (7) Coupling Reaction P2 P1 I I PgNH-CH-CNH-CH-C-CF 2
-C--NR
5
R
6 II I II O (23) OH O Deprotection P2 P1 H2N-CH-CNH-CH-C-CF 2
-C--NR
5
R
6 II I II O (24) OH O Coupling Reaction Oxidation
P
2
P
1 I I
R'
1 C-N H -CH-CNH-CH--C-CF 2 -C-N R 5
R
6 II II II II O O IA O O In Scheme D, step compounds of formula are subjected to a coupling reaction in a manner analogous to that described previously in Scheme A' step with a suitably protected acid of the formula PgNHCH(P 2
)CO
2 H to provide compound of formula (23).
In Scheme D, step compounds of formula (23) are deprotected in a manner analogous to that described in Scheme A, step to provide compounds of formula (24).
In Scheme D, steps and compounds of formula (24) are first subjected to a coupling reaction with an acid of the formula R' 1 C0 2 H in a manner analogous to that described in Scheme A' step and the coupled product is WO 96/02499 PCT/US95/07241 -27then oxidized in a manner analogous to that described in Scheme A' step to provide the compounds of formula IA.
It is understood by one of ordinary skill in the art that compounds of formulas and (15) may be manipulated in a manner analogous to formula in Scheme D, in addition to the manipulations decribed in Schemes A' and A" to provide compounds of formulas IA, IA', IB and IB'.
Having generically described the methods for the preparation of the compounds of this invention, the following specific examples illustrate the chemistry and techniques by which the synthesis may be effected.
The following examples present typical syntheses as described in Schemes A, A" and D. These examples are understood to be illustrative only and are not intended to limit the scope of the present invention in any way. As used herein, the following terms have the indicated meanings: refers to grams; "mmol" refers to millimoles; "ml" refers to milliliters; "bp" refers to boiling point; "mp" refers to melting point; refers to degrees Celsius; "mm Hg" refers to millimeters of mercury; "uL" refers to microliters; "jg" refers to micrograms; "uM" refers to micromolar; "Cbz" means carbobenzyloxy; "DMF" means dimethylformamide; "THF" means tetrahydrofuran; "TBAF" means tetrabutylammonium fluoride; "NMM" means Nmethylmorpholine; "DMSO" means dimethylsulfoxide;
"HOBT"
means hydroxybenzotriazole and "EDC" means 1-(3dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride.
WO 96/02499 WO 9602499PCT/US95/0724 1 -28- Example 1 Preparation of cL,a-Difluoro-Y-[ [[(hydroxylphenylacetyl]amino]-3-methyl-l-oxobutvllamino]- 8-oxo-4-(phenylmethoxy)-N-(Phenylmethyl)-benzenepentanamide.
SOBn OH 0 Hj CF 2
NH
0~ 0 0 Step A Preparation of O-benzyl-N-( tert-butoxycarbonyl)-Ltyrosinal; the starting material in Reaction Scheme A, [Following the procedure of Schirlin, D. and Van Dorsselaer, V. in PCT/US91/09741 published July 23, 1992 with an international publication number of Wa 92/12123.] A mixture of N-tert-butoxycarbonyl-L-0-benzyltyrosine (37.1 g, 100 mmol), dicyclohexylcarbodiimide (20.6 g, 100 mmol), and N-hydroxybenzotriazole hydrate (15.3 g, 100 minol) in anhydrous dichloromethane (350 mL) is stirred at 0 0 C for 10 minutes. To this is added at 0 0 C, N,Odimethylhydroxylamine hydrochloride (9.75 g, 100 mmol) and N-methylmorpholine (10.1 g, 100 mmol). The temperature is allowed to warm to room temperature and stirring is continued for 15 hours. The white precipitate is then filtered off and rinsed with dichloromethane. The filtrate is concentrated under vacuum and the residue is purified by flash chromatography (silica gel, ethyl acetate/cyclohexane, 2:8) to provide the N-tertbutoxycarbonyl-L-Q-benzyltyrosine-N, On-dimethyl-hydroxamate (34.3 g) as a white solid (RF=0.36 in ethyl acetate/cyclohexane, 1:1).
WO 96/02499 PCT/US95/07241 -29- The N-tert-butoxycarbonyl-L-O-benzyltyrosine-N,0dimethyl-hydroxamate (18.2 g, 44 mmol) is dissolved in a mixture of anhydrous diethyl ether/dimethoxyethane (300 mL, 4:1) and cooled to 0°C. To this is added lithium aluminum hydride (1.82 g, 48 mmol) portionwise. The reaction is stirred at 0°C for 1.5 hours. A 1M solution of potassium hydrogen sulfate (55 mL) is then added dropwise with stirring to the reaction. After addition is complete, the aqueous phase is decanted and extracted with ethyl acetate (2 x 200 mL). The combined organic layers are washed with 3N hydrochloric acid (250 mL), water (200 mL), saturated sodium bicarbonate (150 mL) and brine (200 mL). The organic layer is then dried over anhydrous magnesium sulfate, filtered and concentrated under vacuum. The residue is recrystallized from ethyl acetate/pentane to provide N-tert-butoxycarbonyl-L-O-benzyltyrosinal (13 g).
Step B Preparation of 4-tert-butoxycarbonylamino-2,2-difluoro-3hydroxy-5-(4-benzyloxy)phenylpentanoic acid, ethyl ester.
Reaction Scheme A, step To a stirred mixture of N-tert-butoxycarbonyl-L-O-benzyltyrosinal (13.0 g, 36.6 mmol), silver acetate (1.82 g, 10.9 mmol), activated zinc dust (5.02 g, 76.8 mg-atom, washed with 3N hydrochloric acid, water, acetone and ether) and ethyl bromodifluoroacetate (14.8 g, 72.9 mmol) in anhydrous tetrahydrofuran (120 mL) at 0°C is added diethylaluminum chloride (22.4 mL of a 1.8M solution in toluene) over minutes. The temperature is kept below 12 0 C during the addition. The reaction is then allowed to stir at 0°C for minutes and then at room temperature for 4 hours. The reaction is then cooled to 10 0 C and quenched with saturated aqueous ammonium chloride (200 mL). A 1M solution of sodium hydrogen tartrate (200 mL) is added and the reaction is allowed to warm to room temperature. The reaction is filtered and the solids rinsed with ethyl acetate. The filtrate layers are separated and the aqueous layer is WO 96/02499 PCT/US95/07241 extracted with ethyl acetate. The combined organic layers are dried over anhydrous magnesium sulfate, filtered and concentrated under vacuum. The residue is purified by flash chromatography (cyclohexane/ethyl acetate, 4:1) to provide the title compound (8.34 The ratio of diastereomers is approximately 1:1.
Step C Preparation of 4-tert-Butoxycarbonylamino-2,2-difluoro-3hydroxy-5-(4-benzyloxy)phenyl-N-(phenylmethyl)pentanamide.
Reaction Scheme A step To a solution of 4-tertbutoxycarbonylamino-2,2-difluoro-3-hydroxy-5-(4benzyloxy)phenylpentanoic acid, ethyl ester (5.5 g, 11.5 mmol) in anhydrous tetrahydrofuran (50 mL) is added at 0°C, benzylamine (6.15 g, 57.5 mmol). The reaction is stirred for 3 hours at 0°C, then at room temperature for 15 hours.
The reaction is then diluted with ethyl acetate (100 mL), washed with 0.1N aqueous hydrochloric acid (2 x 50 mL), water (50 mL), brine (50 mL) and dried over anhydrous magnesium sulfate. It is then filtered and concentrated under vacuum. The residue is recrystallized from ethyl acetate/pentane to provide the title compound (5.17 g) as a white solid.
Step D Preparation of 4 -amino-2,2-difluoro-3-hydroxy-5-(4benzyloxy)phenyl-N-(phenylmethyl)pentanamide.
Reaction Scheme A, step A solution of 4-tertbutoxycarbonylamino-2,2-difluoro-3-hydroxy-5-(4benzyloxy)phenyl-N-(phenylmethyl)pentamide (5.1 g, 9.4 mmol) in trifluoroacetic acid (100 mL) is stirred at 0°C for 1 hour. The solvent is then removed under vacuum and the residue dissolved in ethyl acetate (100 mL). The organic layer solution is washed with saturated sodium bicarbonate (3 x 50 mL), brine, dried over anhydrous magnesium sulfate, filtered and concentrate under vacuum to provide the title WO 96/02499 PCT/US95/07241 -31compound as a white solid; Rf=0.62 (silica gel, butanol/acetic acid/water, 6:2:2).
Step E Preparation of Dimethylethyl)dimethylsilyl]oxy]phenvlacetyl-L-valine.
To a stirred solution of (R)-(-)-mandelic acid (5.6g, 37 mmol) and tert-butylchlorodimethylsilane (15.0 g, 99.5 mmol) in DMF (100 mL) is added imidazole (16.7 g, 245 mmol) in portions over 15 minutes while maintaining the reaction mixture near 25 0 C with a cool water bath. After 17 hours, the mixture is diluted with water and extracted with two portions of ether. The combined extracts are cooled to 0 C and washed quickly with cold (-10 0 C) 1 N HC1, twice with cold water, dried over anhydrous sodium sulfate, filtered and concentrated under vacuum to provide 15.0 g of a yellow oil which is then dissolved in CH 3 0H (250 mL) and THF (80 mL). A solution of potassium carbonate (14 g, 100 mmol) in water (140 mL) is added with vigorous stirring.
After 1 hour, the mixture is partially concentrated under vacuum and the remaining clear light yellow solution is diluted with brine (300 mL). The cloudy mixture is cooled to -10 0 C and acidified to pH 5 with 1 M KHSO 4 Extraction with cold (-10 0 C) ether and washing of the extracts with two portions of brine provides, after drying with anhydrous sodium sulfate, filtration and concentration under vacuum, 6.6 g of (R)-O-tert-butyldimethyl-silylmandelic acid as a yellow oil. This crude silylated acid (25 mmol) is dissolved in CH 3 CN (80 mL) and the resulting solution is cooled to -10 0 C. To the stirred solution is added NMM (2.52 g, 24.9 mmol), then isobutylchloroformate (3.72 g, 27.2 mmol). After 10 minutes, a solution of L-valine methyl ester hydrochloride (7.32 g, 43.7 mmol) and NMM (4.08 g, 40.3 mmol) in H 2 0 (20 mL) is added and stirring is continued at -10 0 C for 20 minutes. The reaction mixture is poured into ice-cold dilute HC1 containing some NaCl and extracted with two portions of ethyl acetate. The combined extracts WO 96/02499 PCTIUS95/07241 -32are washed with water, aqueous saturated sodium bicarbonate, brine, dried over anhydrous magnesium sulfate, filtered and concentrated under vacuum. Two flash chromatographies (12% ethyl acetate in cyclohexane) provide 4.4 g of coupled silylated ester (Rf 0.4, 4/1 cyclohexane/ethyl acetate) contaminated with Nisobutyloxycarbonyl-L-valine methyl ester. The silylated ester (4.4 g, 12 mmol) is dissolved in THF (15 mL) with stirring and a solution of 1 M tetrabutylammonium fluoride (TBAF) in THF (11.6 mL) is added. After 1.25 hours, the solution is concentrated under vacuum and the residue is purified by flash chromatography (4/1 cyclohexane/ethyl acetate) to provide 2.4 g (37% overall from O-tertbutyldimethylsilylmandelic acid) of N-[(R)-mandeloyl]-Lvaline methyl ester as an oil. Following the procedures in a manner analogous to that described above, the material is reconverted to its O-tert-butyldimethylsilyl ether in 82% yield after flash chromatography (12% ethyl acetate in cyclohexane). To a stirred solution of this ester (13.5 mmol) in CH 3 OH (125 mL) and water (5 mL) is added LiOH*H 2 0 (0.59 g, 14 mmol). After 17 hours an additional amount of LiOH.H 2 0 (0.06 g) is added and stirring is continued for 2 hours. The solution is concentrated under vacuum and the residue is diluted with water (100 mL) and washed with ether. The aqueous layer is acidified with ice-cold 6 N HC1 and extracted with two portions of ether. The combined extracts are washed with water, dried over anhydrous magnesium sulfate, filtered and concentrated under vacuum to provide the title compound in 48% yield as a white solid: mp 146-148 0 C; IR (CHC13) Vmax 2957, 2932, 1718, 1678, 1516, 1096, 863, 840 IH NMR (CDC3) 6 9.17 (bs, 1 7.51 1 H, J=9.0 Hz), 7.43-7.40 2 7.32- 7.25 3 5.15 1 4.51 (dd, 1 H, J=9.0, 4.3 Hz), 2.34-2.28 1 1.00 3 H, J=6.9 Hz), 0.97 (d, 3 H, J=7.0 Hz), 0.94 9 0.11 3 -0.08 3 13 C NMR (CDC13) 6 175.49, 172.65, 139.36, 128.32, 128.21, 126.68, 75.66, 56.60, 31.04, 25.66, 19.01, 18.02, WO 96/02499 PCT/US95/07241 -33- 17.44, -4.83, -5.31; mass spectrum, m/z 394 (M 29), 366 (M 1, 100), 350, 308; exact mass calcd for Cg9H3 2
NO
4 Si 366.2101, found 366.2099; [a] 20 D -56.60 (c 1.00, Anal. Calcd for C 19
H
31
NO
4 Si: C, 62.43; H, 8.55; N, 3.83.
Found: C, 62.22; H, 8.61; N, 3.65.
Step F Preparation of dimethylethyl)dimethylsilyl]oxylphenylacetyllamino]-3methyl-l-oxobutyl]amino]-2,2-difluoro-5-[4- (phenylmethoxy)phenyl]-N-(phenylmethyl)-L-qlyceropentanamide.
Reaction Scheme step dimethylethyl)dimethylsilyl]oxy]phenylacetyl]-L-valine prepared above (25 mmol) is dissolved in CH 3 CN (80mL) and the resulting solution cooled to -10 0 C. To the stirred solution is added N-methylmorpholine (2.52 g, 24.9 mmol, NMM), followed by isobutylchloroformate (3.72 g, 27.2 mmol). After 10 minutes, a solution of 4-amino-2,2difluoro-3-hydroxy-5-(4-benzyloxy)phenyl-N-(phenylmethyl) pentanamide (43.7 mmol, prepared in step above) and NMM (4.08 g, 40.3 mmol) in H 2 0 (20 mL) is added and stirring is continued at -10 0 C for 20 minutes. The reaction mixture is poured into ice-cold dilute HC1 containing some NaC1 and is extracted with two portions of ethyl acetate. The combined extracts are washed with water, aqueous saturated sodium bicarbonate, brine, dried over anhydrous magnesium sulfate, filtered and concentrated under vacuum. The title compound is obtained in 76% yield as a clear colorless oil which crystallizes on standing after flash chromatography (6% acetone in CH 2 C1 2 mp 117-120 0 C; IR (KBr) vax 3418, 1704, 1677, 1653, 1540, 1513, 1246, 1095, 839 cm- 1 1 H NMR (CDC1 3 6 7.44-7.18 16 6.96 3 H, J=8.7 Hz), 6.84 2 H, J=8.7 Hz), 6.10 1 H, J=8.1 Hz), 5.10 1 5.01 2 4.69 1 H, J=6.9 Hz), 4.43 (dd, 1 H, J=14.7, Hz), 4.34 (dd, 1 H, J=14.7, 6.0 Hz), 3.96 (dd, 1 H, 6.9 Hz), 3.91-3.82 2 2.76 (dd, 1 H, J=13.8, WO 96/02499 PCT/US95/07241 -34- 7.8 Hz), 2.66 (dd, 1 H, J=13.8, 7.8 Hz), 2.22-2.11 1 0.94 9 0.88 3 H, J=6.6 Hz), 0.86 3 H, J=6.9 Hz), 0.09 3 -0.06 3 13 C NMR (CDC1 3 6 173.37, 171.91, 163.92 J=35 Hz), 159.01, 139.62, 137.40, 137.31, 130.47, 129.60, 129.16, 128.94, 128.80, 128.70, 128.37, 128.31, 128.22, 127.83, 126.63, 115.50 (t, J=257 Hz), 115.36, 76.21, 70.67 J=25 Hz), 70.37, 58.79, 51.93, 43.76, 36.52, 29.97, 26.08, 19.80, 18.47, 17.96, 4.41, -4.89; 19F NMR (CDC13) 6 -118.40 -118.45 mass spectrum, m/z 816 29), 788 (M 573, 469, 441, 423 (100); exact mass calcd for C 44
H
56
F
2
N
3 06Si 788.3906, found 788.3932; [a] 20 D -74.6 (c 1.02, CH30H). Anal. Calcd for C 44
H
55
F
2
N
3 06Si: C, 67.06; H, 7.04; N, 5.33. Found: C, 67.07; H, 7.14; N, 5.18.
Step G Preparation of final title compound.
Reaction Scheme steps and The above prepared alcohol is oxidized under Swern conditions as follows: To a stirred solution of 2 M oxalyl chloride/CH 2 Cl2 at -60 0 C under nitrogen is added dropwise anhydrous DMSO (0.42 mL, 5.9 mmol). After 10 minutes, a solution of [3 dimethylethyl)dimethylsilyl]oxy]phenylacetyl]amino]-3methyl-l-oxobutyl]amino]-2,2-difluoro-5-[4- (phenylmethoxy)phenyl]-N-(phenylmethyl)-L-glyceropentonamide (0.50 mmol) in CH 2 C1 2 (3 mL) and anhydrous DMSO mL) is added. The solution is allowed to stir at to -55 0 C for 5 hours. It is then warmed to -20 0 C, followed by addition of triethylamine (1.39 mL, 10.0 mmol) over minutes. The reaction mixture is then allowed to warm to room temperature overnight. It is then diluted with ethyl acetate (60 mL) and filtered, washing the solids with ethyl acetate. The filtrate is washed with 0.1 M HC1 (3 x mL), brine, dried over anhydrous magnesium sulfate, filtered and concentrated under vacuum to provide 375 mg of crude ketone. This is dissolved in 4/1 acetic acid/H20 WO 96/02499 PCT/US95/07241 mL). The stirred solution is heated at 80 0 C for 19 hours, allowed to cool to room temperature before being diluted with water and extracted with two portions of CH 2 C12. The combined extracts are washed with water, brine, dried over anhydrous magnesium sulfate, filtered and concentrated under vacuum. Flash chromatography of the residue acetone in CH 2 C12) provides 180 mg title compound contaminated by residual silylated compound. The material is resubjected to the reaction conditions and purified by flash chromatography to provide 121 mg of final title compound. Two recrystallizations from ethyl acetate/pentane provide 30 mg of final title compound as a pale yellow solid: mp 144-150 0 C; IR (KBr) vmax 3398, 3300, 1683, 1656, 1540, 1513, 1454, 1244, 698 cm- 1 1 H NMR (DMSO-d 6 6 9.77-9.72 1 8.69 1 H, J=7.6 Hz), 7.80 and 7.73 (2d, 1 H total, J=9.3 Hz), 7.45-7.21 (m, 7.12 and 7.10 (2d, 2 H total, J=8.7 Hz), 6.89 and 6.87 (2d, 2 H total, J=8.8, 8.4 Hz), 6.21 and 6.20 (2d, 1 H total, J=5.1 Hz), 5.05 2 5.02 and 5.00 (2s, 1 H total), 5.03-4.90 1 4.38 2 H, J=5.7 Hz), 4.23- 4.17 1 3.14-3.07 and 2.65-2.57 2 H total), 1.97-1.88 and 1.78-1.73 1 H total), 0.73 and 0.71 (2d, 3 H total, J=6.7 Hz), 0.57 and 0.49 (2d, 3 H total, J=6.9, 6.7 Hz); 19 F NMR (DMSO-d 6 6 -110.31 J=268 Hz), -110.54 J=268 Hz), -112.21 J=268 Hz), -112.25 J=269 Hz); mass spectrum, m/z 700 (M 29), 672 (M 1, 100), 652, 439, 421, 402; exact mass calcd for C 38
H
40
F
2
N
3 0 6 672.2885, found 672.2842.
WO 96/02499 PCT/US95/07241 -36- Example 2 Preparation of 9R*, 13S*)1-4,4,-Difluoro-9-(1methylethyl)-3,5,8,11-tetraoxo-1,13-diphenyl-6-[[4- (phenylmethoxy)phenyl]methyl]-12-oxa-2,7,10triazatetradecan-14-oic Acid, Methyl Ester.
OBn H CF 2
NH
CH
3 0 2 C O O O Step A Preparation of [3f,4(S)]-2,4,5-Trideoxy-4-[[2-[[(l,1- [4-(phenylmethoxy)phenyl]-N-(phenylmethyl)-L-glyceropentanamide.
Reaction Scheme D, step To an ice-cold stirred solution of 4-amino-2,2-difluoro-3-hydroxy-5-(4benzyloxy)phenyl-N-(phenylmethyl)pentanamide (556 mg, 1.23 mmol, prepared in example 1, step D) in 2:1 dry CH 2 Cl 2
/DMF
(13.5 mL) is added HOBT (0.19 g, 1.2 mmol), DCC (0.26g, 1.3 mmol) and N-t-Boc-L-valine (0.27 g, 1.2 mmol). The reaction mixture is allowed to warm to 25 0 C overnight, then diluted with ethyl acetate/cyclohexane, washed with dilute aqueous sodium bicarbonate, water, brine, dried over anhydrous magnesium sulfate, filtered and concentrated under vacuum to provide a gummy solid. This is triturated with CH 2 C1 2 and filtered to remove some of the dicyclohexylurea. The filtrate is concentrated under vacuum and the residue is purified by flash chromatography (3:2 cyclohexane/ethyl acetate) to provide 694 mg of the title compound as an ivory solid. The major 3 diastereomer is separated in 69% yield by recrystallization from ether/CH 2 Cl 2 WO 96/02499 PCT/US95/07241 -37- Step B Preparation of the chloroformate of formula: 0 CI
CH
3 0 2 C O To a stirred solution of triphosgene (6.65 g, 22.4 mmol) in CH 2 C1 2 (25 mL) under nitrogen is added methyl mandelate (5.36 g, 32.3 mmol) followed by the rapid addition of a solution of pyridine (2.66 mL, 32.9 mmol) in
CH
2 C1 2 (2.5 mL). The reaction mixture is warmed to reflux and pyridinium chloride separated. After 17 hours at room temperature, the reaction mixture is partially concentrated under vacuum, diluted with ethyl acetate, and filtered.
The filtrate is concentrated concentrated under vacuum and the residue is again dissolved in ethyl acetate and filtered. Concentration under vacuum provides 7.25 g of a pale yellow oil composed of approximately a 20:1:1 mixture of chloroformate: carbonate: methyl mandelate. For the chloroformate IR (neat) Vmax 1772, 1750 cm-; 1 H NMR (CDC1 3 6 7.5-7.37 5 5.98 1 3.78 3 H).
Step C Preparation of 9R*, 13S*)]-4,4,-Difluoro-5hydroxy-9-(l-methylethyl)-3,8,11-trioxo-l,13-diphenyl-6- [[4-(phenylmethoxy)phenyl]methyl]-12-oxa-2,7,10triazatetradecan-14-oic Acid, Methyl Ester.
Reaction Scheme D, steps and A flask containing the major diastereomer of Trideoxy-4-[[2-[[(1,1-dimethylethoxy)carbonyl]amino]-3methyl-l-oxobutyl]amino-5-[4-(phenylmethoxy)phenyl]-N- (phenylmethyl)-L-glycero-pentonamide (403 mg, 0.630 mmol) is submersed in an ice bath and ice-cold trifluoroacetic acid (TFA) (5 mL) is added with stirring. After minutes, the solution is concentrated under vacuum at 25 0
C
and the residue is carefully partitioned between ethyl acetate and aqueous saturated sodium bicarbonate. The WO 96/02499 PCT/US95/07241 -38organic layer is separated, washed with brine, dried over anhydrous magnesium sulfate, filtered and concentrated under vacuum to provide 334 mg of deprotected amine which is immediately suspended in CH 2 C12 (5 mL) at 0°C and treated with NMM (77 pL, 0.70 mmol) and the chloroformate prepared above (160 mg, 0.70 mmol) dissolved in CH 2 C1 2 (1 mL). After stirring for 1.5 hours, the reaction mixture is poured into ethyl acetate/dilute aqueous ammonium chloride.
The organic layer is separated, washed with dilute aqueous sodium bicarbonate, brine, dried over anhydrous magnesium sulfate, filtered and concentrated under vacuum. Flash chromatography (4:1 CH 2 Cl 2 /ethyl acetate) of the residue provides 332 mg of a single diastereomer of the title compound as a white foam: 1 H NMR (CDCl 3 6 7.44-7.26 7.12 2 H, J=8.4 Hz), 6.89 2 H, J=8.4 Hz), 6.53 1 H, J=8.4 Hz), 5.89 1 5.40 1 H, J=7.9 Hz), 5.02 2 4.70 1 H, J=7.0 Hz), 4.49 (apparent d, 2 H, J=5.8 Hz), 4.3-4.1 2 3.85 (dd, 1 H, J=7.8, Hz), 3.63 3 3.05-2.87 2 2.2-2.04 1 H), 1.90 1 0.83 3 H, J=6.7 Hz), 0.71 3 H, J=6.8 Hz); 19 F NMR (CDC1 3 6 -116.60 (dd, 1 F, J=259, 9 Hz), 118.73 (dd, 1 F, J=259, 15 Hz).
Step D Preparation of final title compound.
Reaction Scheme step The above prepared alcohol is subjected to Swern Oxidation conditions in a manner analogous to that described previously in example 1, step G. The residue is purified by flash chromatography (4:1 CH 2 Cl 2 /ethyl acetate) to provide 52% of the final title compound. Precipitation with pentane from an ethyl acetate solution provides 29% of final title compound as a white powder: IR (KBr) Vmax 3399, 3316, 1745, 1699, 1668, 1534, 1513, 1241 cm-l; 1 H NMR (CDC1 3 6 5.89, 5.87, and 5.82 (3s in 6.0:1.8:1.0 ratio, 1 5.05 and 5.03 (2s in 1:1.6 ratio, 2 3.69, 3.67, and 3.65 (3s in 2.5:1.0:1.9 ratio, 3 1 9 F NMR (CDC13) 6 -112.53 -112.60 -116.25 WO 96/02499 PCT/US95/07241 -39- J=256 Hz), -117.00 J=256 Hz), -120.21 J=256 Hz), -120.78 J=256 Hz), d ratio mass spectrum, m/z 770 (M 41), 758 (M 29), 731, 730 (M 1, 100); exact mass calcd for C 40
H
42
F
2
N
3 0 8 730.2940, found 730.2925; [a] 2 0 D -48.3° (c 1.22, CH 3 0H). Anal. Calcd for
C
40
H
41
F
2
N
3 0 8 C, 65.83; H, 5.66; N, 5.76. Found: C, 65.16; H, 5.69; N, 5.79.
Example 3 Preparation of a,a-Difluoro-y-[[3-methyl-l-oxo-2-[[(1oxo-trans-3-phenyl-2-propenyl)amino]butyl]amino]-8-oxo-4- (phenylmethoxy)-N-(phenylmethyl)-benzene-pentanamide.
SOBn N H H CF2 NH OO
O
Step A Preparation of N-(trans-Cinnamoyl)-L-valine To a stirred solution of L-valine methyl ester hydrochloride (1.97 g, 11.8 mmol) and NMM (1.10 g, 10.8 mmol) in CH 2 C1 2 (15 mL) and DMF (20 mL) is added a solution of 1-trans-cinnamoylimidazole (2.15 g, 10.8 mmol) in CH 2 C12 (28 mL). After 18 h, additional L-valine methyl ester hydrochloride (0.36 g, 2.1 mmol) is added and the reaction 3 mixture is stirred for 5 h before being diluted with water (100 mL) and extracted with CHC1 3 The organic extract is washed with three portions of water, dried (MgSO 4 and concentrated in vacuo. Filtration through a short silica gel column (1/1 ethyl acetate/cyclohexane) provides 2.0 g of N-(trans-cinnamoyl)-L-valine methyl ester. To a stirred solution of this ester (13.5 mmol) in CH30H (125 mL) and water (5 mL) is added LiOH.H 2 0 (0.59 g, 14 mmol).
After 17 hours an additional amount of LiOH.H 2 0 (0.06 g) is WO 96/02499 PCT/US95/07241 added and stirring is continued for 2 hours. The solution is concentrated in vacuo and the residue is diluted with water (100 mL) and washed with ether. The aqueous layer is acidified with ice-cold 6 N HC1 and extracted with two portions of ether. The combined extracts are washed with water and dried (MgSO4). Concentration in vacuo provides 67% of the title compound as a white solid: mp 63-68 0 C; IR (CHC13) Vmax 3008, 2970, 1718, 1670, 1629, 1512, 1196, 667 cm- 1 1 H NMR (CDC13) 6 9.94 (bs, 1 7.65 1 H, J=15.6 Hz), 7.50-7.46 2 7.35-7.31 3 6.56 1 H, J=8.7 Hz), 6.52 1 H, J=15.6 Hz), 4.75 (dd, 1 H, J=8.7, 4.8 Hz), 2.34-2.28 1 1.02 3 H, J=6.9 Hz), 0.99 3 H, J=6.6 Hz); 13 C NMR (CDC13) 6 175.31, 166.66, 142.42, 134.50, 129.93, 128.79, 127.94, 119.75, 57.47, 31.19, 19.00, 17.78; mass spectrum, m/z 276 (M 29), 248 (M 203, 131 (100); exact mass calcd for C 14
H
18
NO
3 248.1287, found 248.1290; [a]20 D +24.10 (c 1.02, CH 3 0H).
Anal. Calcd for C 14
H
17
NO
3 *0.1H 2 0: C, 67.51; H, 6.96; N, 5.62. Found: C, 67.30; H, 7.10; N, 5.57.
Step B Preparation of [3E,4(S)]-2,4,5-Trideoxy-2,2-difluoro-4-[[3methyl-l-oxo-2-[(l-oxo-trans-3-phenyl-2propenyl)amino]butyl]amino]-5-[4-(phenylmethoxy)phenyl]-N- (phenylmethyl)-L-glycero-pentanamide.
Reaction Scheme step N-(trans-Cinnamoyl)-Lvaline prepared above (25 mmol) is dissolved in CH 3 CN and the resulting solution cooled to -10 0 C. To the stirred solution is added N-methylmorpholine (2.52 g, 24.9 mmol, NMM), followed by isobutylchloroformate (3.72 g, 27.2 mmol). After 10 minutes, a solution of 4-amino-2,2difluoro-3-hydroxy-5-(4-benzyloxy)phenyl-N-(phenylmethyl) pentanamide (43.7 mmol, prepared in example 1, step D and NMM (4.08 g, 40.3 mmol) in H 2 0 (20 mL) is added and stirring is continued at -10 0 C for 20 minutes. The reaction mixture is poured into ice-cold dilute HC1 containing some NaCl and is extracted with two portions of ethyl acetate. The WO 96/02499 PCT/US95/07241 -41combined extracts are washed with water, aqueous saturated sodium bicarbonate, brine, dried over anhydrous magnesium sulfate, filtered and concentrated under vacuum. The residue is recrystallized from ethyl acetate/pentane to provide 61% of a 2:1 mixture of diastereomers of title compound as a light tan solid: mp 222-227 0 C; IR (KBr) vmax 3410, 3287, 1682, 1652, 1620, 1540, 1512, 1243, 1217, 697 cm-1; 1 H NMR (DMSO-d 6 6 9.17-9.11 1 8.05 and 8.01 (2d, 1 H total, J=9.1, 9.1 Hz), 7.85 and 7.71 (2d, 1 H total, J=9.1, 9.0 Hz), 7.59-7.22 16 7.12 2 H, Hz), 6.94-6.85 3 6.26 1 H, J=7.6 Hz), 5.04 and 4.96 (2s, 2 H total), 4.42-3.99 5 2.79- 2.63 2 2.09-1.98 and 1.92-1.85 (2m, 1 H total), 0.86 and 0.74 (2d, 3 H total, J=6.9 Hz), 0.83 and 0.64 (2d, 3 H total, J=6.9 Hz); 19 F NMR (DMSO-d 6 6 -111.11 (dd, J=255, 6 Hz), -112.12 (dd, J=254, 8 Hz), -120.35 (dd, J=254, 19 Hz), -122.04 (dd, J=255, 20 Hz); mass spectrum, m/z 698 (M 29), 670 (M 1, 100), 650, 441, 423. Anal.
Calcd for C 3 9H 41
F
2
N
3 0 5 C, 69.94; H, 6.17; N, 6.27. Found: C, 68.59; H, 6.32; N, 5.90.
Step C Preparation of the final title compound.
Reaction Scheme step The above prepared alcohol is oxidized under Swern conditions as follows: To a stirred solution of 2 M oxalyl chloride/CH 2 Cl 2 (2.0mL) at 0 C under nitrogen is added dropwise anhydrous DMSO (0.42 mL, 5.9 mmol). After 10 minutes, a solution of the above prepared alcohol (0.50 mmol) in CH 2 C1 2 (3 mL) and anhydrous DMSO (1.5 mL) is added. The solution is allowed to stir at to -55 0 C for 5 hours. It is then warmed to -20 0
C,
followed by addition of triethylamine (1.39 mL, 10.0 mmol) over 5 minutes. The reaction mixture is then allowed to warm to room temperature overnight. It is then diluted with ethyl acetate (60 mL) and filtered, washing the solids with ethyl acetate. The filtrate is washed with 0.1 M HC1 (3 x 15 mL), brine, dried over anhydrous magnesium sulfate, WO 96/02499 PCT/US95/07241 -42filtered and concentrated under vacuum. Flash chromatography acetone in CH 2 C1 2 followed by recrystallization from ethyl acetate/pentane provides 34% of the final title compound as a white solid: mp 187-194C; IR (KBr) vmax 3403, 3285, 1691, 1654, 1623, 1540, 1513, 1454, 1243, 1219, 1178, 697 cm-l; 1 H NMR (DMSO-d 6 6 9.77- 9.68 1 8.67-8.62 (m 1 8.05-8.00 1 7.56 2 H, J=7.0 Hz), 7.50-7.21 14 7.14 2 H, Hz), 6.91-6.83 3 5.04-4.95 3 4.42- 4.33 3 3.14-3.05 1 2.74-2.61 1 H), 2.02-1.76 1 0.84 and 0.81 (2d, 3 H total, 6.9 Hz), 0.64 and 0.59 (2d, 3 H total, J=7.1, 6.9 Hz); 19
F
NMR (DMSO-d 6 6 -110.26 J=269 Hz), -110.53 J=268 Hz),-112.03 J=269 Hz), -112.04 J=268 Hz)-major diastereomer pair; -105.07 J=265 Hz), -105.08 (d, J=266 Hz), -110.59 J=265 Hz), -110.61 J=266 Hz)minor diastereomer pair; mass spectrum, m/z 696 29), 668 (M 439, 230, 202, 131, 124, 91 (100); exact mass calcd for C 39
H
40
F
2
N
3 05 668.2936, found 668.2953; [a] 20
D
+7.230 (c 0.622, DMSO-d 6 Anal. Calcd for C 39
H
39
F
2
N
3 0 5
C,
70.15; H, 5.89; N, 6.29. Found: C, 69.63; H, 5.85; N, 6.01.
WO 96/02499 PCTIUS95/07241 -43- Example 4 Preparation of a,a-Difluoro-y-[[2-[2-hydroxy-l-oxo-4- (phenylbutyl)amino]-3-methyl-l-oxobutyl]amino]-B-oxo-4- (phenylmethoxy)-N-(phenylmethyl)-benzene-pentanamide.
sOBn OH 0 1 NH H CF 2
NH
H II o o o Step A Preparation Dimethylethyl)dimethylsilyl]oxy]-4-phenylbutyryl]-L-valine.
(R)-(-)-2-hydroxy-4-phenylbutyric acid is coupled with L-valine methyl ester hydrochloride under standard coupling conditions in a manner analogous to that described previously, such as example 2, step A, to provide hydroxy-4-phenylbutyryl)-L-valine methyl ester as a yellow solid in 89% yield. Treatment with tert-butyldimethylsilyl chloride in a manner analogous to that described previously in example 1 for the preparation of dimethylethyl)dimethylsilyl]oxy]phenylacetyl]-L-valine provides the ester of the title compound as a colorless oil in 83% yield after flash chromatography (9/1 cyclohexane/ethyl acetate). The ester is hydrolyzed with LiOH*H 2 0 in a manner analogous to that described previously in example 1 for the preparation of dimethylethyl)dimethylsilyl]oxy]phenylacetyl]-L-valine to provide the title compound in 58% yield as a white solid: mp 110-113 0 C; IR (KBr) Vmax 3387, 2958, 2932, 2897, 2860, 1719, 1626, 1531, 1251, 1097, 839 cm-l; H NMR (CDC13) 6 7.24-7.12 7 4.54 (dd, 1 H, J=8.7, 4.5 Hz), 4.32 (t, 1-H, J=4.8 Hz), 2.76-2.55 2 2.36-2.25 1 H), 2.19-2.07 1 2.02-1.90 1 1.00 3 H, J=7.2 Hz), 0.97 3 H J=6.9 Hz), 0.97 9 0.13 3 H), 0.10 3 13 C NMR (CDC1 3 6 175.64, 174.08, 141.69, WO 96/02499 PCT/US95/07241 -44- 128.42, 128.30, 125.77, 72.73, 59.64, 36.98, 30.75, 30.02, 25.67, 19.05, 17.93, 17.54, -4.79, -5.15; mass spectrum, m/z 394 (M 393 378, 336 (100), 289, 117, 91, 73; [a]20D +13.70 (c 1.00, CH 3 0H). Anal. Calcd for
C
21
H
35
NO
4 Si: C, 64.08; H, 8.96; N, 3.56. Found: C, 63.87; H, 9.07; N, 3.45.
Step B Preparation of dimethylethyl)dimethylsilyl]oxyl-l-oxo-4phenylbutyl]amino]-3-methyl-l-oxobutyl]aminol-5-[4- (phenylmethoxy)phenyl]-N-(phenylmethyl)-L-qlyceropentanamide.
Reaction Scheme step Dimethylethyl)dimethylsilyl]oxy]-4-phenylbutyryl]-L-valine is coupled with 4-amino-2,2-difluoro-3-hydroxy-5-(4benzyloxy)phenyl-N-(phenylmethyl)pentanamide prepared in example 1, step D under standard coupling conditions in a manner analogous to that described previously, such as in example 2, step A, to provide the title compound after flash chromatography acetone in CH 2 C12) as a tan foam in 61% yield: mp 62-67 0 C; 1 H NMR (CDC13) 6 7.44-7.11 17 H), 7.04 2 H, J=8.7 Hz), 6.86 2 H, J=8.7 Hz), 6.11 (d, 1 H, J=8.1 Hz), 5.01 2 4.71 1 H, J=6.9 Hz), 4.52 (dd, 1 H, J=14.6, 6.5 Hz), 4.35 (dd, 1 H, J=14.4, 5.4 Hz), 4.28 1 H, J=4.8 Hz), 4.15-3.93 3 2.91 (d, 2 H, J=7.8 Hz), 2.72 (ddd, 1 H, J=13.2, 12.0, 4.8 Hz), 2.57 (ddd, 1 H, J=13.5, 12.3, 5.4 Hz), 2.15-1.90 3 0.98 9 0.87 3 H, J=2.4 Hz), 0.85 3 H, J=2.1 Hz), 0.14 3 0.10 3 19 F NMR (CDC13) 6 -117.17 (dd, J=259, 15 Hz), -118.62 (dd, J=259, 10 Hz); mass spectrum, m/z 844 29), 816 758, 469, 441, 423 (100), 91; [a] 20 D -30.60 (c 0.547, CH 3 0H). Anal. Calcd for
C
4 6
H
5 9
F
2
N
3 06Si: C, 67.70; H, 7.29; N, 5.15. Found: C, 67.32; H, 7.22; N, 5.15.
WO 96/02499 PCT/US95/07241 Step C Preparation of final title compound.
Reaction Scheme steps and The above prepared alcohol is oxidized under Swern conditions in a manner analogous to that described previously in example 1, step G to provide the protected ketone as a light brown oil in 72% yield after flash chromatography acetone in
CH
2 C1 2 Deprotection of the ketone (217mg) in a manner analogous to that described previously in example 1, step G provides the title compound after flash chromatography (6, and finally 15% acetone in CH 2 C1 2 104 mg as a light yellow glass. Crystallization from CH2Cl2/ether and three recrystallizations from ethyl acetate/pentane provides 34 mg of title compound as a white solid: IR (KBr) Vmax 3394, 3300, 1686, 1651, 1534, 1513, 1498, 1243, 698 cm- 1 1 H NMR (DMSO-d 6 6 9.76-9.69 1 H) 8.70-8.66 1 7.52-7.11 19 6.88 and 6.87 (2d, 2 H total, J=8.7 Hz), 5.73 1 H, J=5.8 Hz), 5.03 and 5.00 (2s overlapping m, 3H total, J=8.3 Hz), 4.35 (apparent d, 2 H, J=6.1 Hz), 4.26-4.20 1 3.94-3.87 1 3.15- 3.06 1 2.73-2.58 2 1.96-1.66 3 0.77 and 0.76 (2d, 3 H total, J=6.7 Hz), 0.63 and 0.53 (2d, 3 H total, J=6.8, 6.6 Hz); 19 F NMR (DMSO-d 6 6 -110.35 J=268 Hz), -110.47 J=268 Hz), -112.14 J=268 Hz), -112.20 J=268 Hz); mass spectrum, m/z 700 (M 680, 622, 439 (100), 421, 254, 91.
WO 96/02499 PCT/US95/07241 -46- Example Preparation of N-[l-[[[3,3-Difluoro-4-[[2-methyl-1- [(phenylmethoxy)methyl]-propyl]amino]-2,4-dioxo-l-[(4- (phenylmethoxy)phenyl]-methyl]butyl]amino]carbonyl]-2methylpropyl]-B-oxo-4-morpholinepropanamide.
H OBn 0 0 N NH C F NH H
O
Bn o o O A Step A Preparation of [1 4(S)]-2,4,5-Trideoxy-4-[[(1,1dimethylethoxy)-carbonyllamino]-2,2-difluoro-N-[2-methyl-l- [(phenylmethoxy)methyl]propyl]-5-[4-(phenylmethoxy)phenyl]- L-glycero-pentanamide.
Reaction Scheme A, step To a stirred solution of O-benzyl-D-valinol (5.42 g, 28.0 mmol) in CH 2 C12 (15 mL) is added a 2 M solution of (CH3)3Al/toluene (14.0 mL, 28.0 mmol) over 20 minutes. During the addition more CH 2 C1 2 mL) is added to convert the resulting paste to a thin slurry. As the addition proceeds, a solution again forms with vigorous gas evolution as it warms to reflux. After the addition is complete, the solution is allowed to stir at reflux for 15 minutes as gas evolution subsides. The solution is allowed to cool to room temperature and a solution of 4-tert-butoxycarbonylamino-2,2-difluoro-3hydroxy-5-(4-benzyloxy)phenylpentanoic acid, ethyl ester (5.38 g, 11.2 mmol, prepared in example 1) in CH 2 C12 (20 mL) is added over 15 minutes with moderate gas evolution. The resulting solution is heated at reflux overnight. The reaction mixture is cooled to 0°C and carefully quenched with excess methanol. The resulting solution is washed with 1 N HC1. The aqueous layer is extracted with ether.
The combined organic extracts are washed with water, dried over anhydrous magnesium sulfate, filtered and concentrated WO 96/02499 PCT/US95/07241 -47under vacuum to provide after flash chromatography (4:1 cyclohexane/ethyl acetate) 5.5 g of title compound as a beige powder.
Step B Preparation of [1 4 (S)]-4-Amino-2,4,5-trideoxy- 2,2-difluoro-N-[2-methyl-l-[(phenylmethoxy)methyl]propyl]- 5-[4-(phenylmethoxy)phenyl]-L-qlycero-pentanamide.
Reaction Scheme A, step A solution of the above prepared alcohol (3.6 g, 5.7 mmol) in HCO 2 H (50 mL) is allowed to stir at room temperature for 3 hours. The solution is then concentrated under vacuum at 35 0 C. The residue is dissolved in ethyl acetate and aqueous sodium bicarbonate is added with vigorous stirring. The layers are separated and the organic layer is washed with water.
The combined aqueous layers are again extracted with ethyl acetate. The combined organic extracts are dried anhydrous magnesium sulfate, filtered and concentrated under vacuum to provide 3.0 g of the title compound as an orange oil.
Step C PreTaration of [1 4(S)1-2,4,5-Trideoxy-4-[[2- [[(l,l-dimethylethoxy)carbonyl]amino]-3-methyl-loxobutyl]amino]-2,2-difluoro-N-[2-methyl-l- [(phenylmethoxy)methyl]propyl]-5-[4-(phenylmethoxy)phenyl]-L-qlycero-pentanamide.
Reaction Scheme D, step The above prepared amine is coupled under standard conditions in a manner analogous to that described previously, such as exmaple 2, step A, with N-(tert-butoxycarbonyl)-L-valine (1.21 g, 5.61 mmol) to provide, after flash chromatography (9:1 CH 2 Cl 2 /ethyl acetate), 2.3 g of title compound as a yellow powder.
WO 96/02499 PCT/US95/07241 -48- Step D Preparation of [1 3 4(S)]-4-[[2-Amino-3-methyl-loxobutyl]amino]-2,4,5-trideoxy-2,2-difluoro-N-[2-methyl-- [(phenylmethoxy)methyl]-propyl]-5-[4- (phenylmethoxy)phenyl]-L-qlycero-pentanamide.
Reaction Scheme D, step(b): A solution of the above prepared amide (2.0 g, 2.8 mmol) in HCO 2 H (25 mL) is allowed to stir at room temperature for 6 hours. The solution is concentrated under vacuum at 30 0 C and the residue is dissolved in ethyl acetate. Aqueous sodium bicarbonate is added with vigorous stirring. White solids precipitate out which are filtered and found to be the formate salt of the (S)-amino alcohol. The organic layer of the filtrate is separated, washed with water, dried over anhydrous magnesium sulfate, filtered and concentrated under vacuum to provide an amber oil containing some white solids; both the and (S)-amino alcohol and the lactam by-product are present. The residue is slurred in CHC13 for 1 hour and the precipitated solids, again the formate salt of the amino alcohol, are collected. The filtrate is concentrated under vacuum and the residue dissolved in CH 2 C12 and additional formate salt of the (S)-amino alcohol is collected. Flash chromatography of the residual oil (5:1
CH
2 Cl 2 /ethyl acetate to remove the lactam, then 9:1 CH 2 C12/
CH
3 0H) provides a yellow semi-solid which is the (R)-amino alcohol contaminated with 5% of the formate of the amino alcohol. Yield: 800 mg of the formate salt of the (S)-amino alcohol and 640 mg of the (R)-amino alcohol. To a stirred suspension of the formate salt of the (S)-amino alcohol (800 mg, 1.19 mmol) in ethyl acetate is added 10% aqueous sodium bicarbonate. After 2 hours at room temperature, the solids dissolve. The organic layer is separated, washed with water, dried over anhydrous magnesium sulfate, filtered and concentrated under vacuum to provide 712 mg of title compound as a white powder: mp 160-164 0 C; IR (KBr) umax 3383, 3327, 1676, 1632, 1530, 1514, 1244, 1109 cm- 1 1 H NMR (CDC13) 6 7.73 1 H, WO 96/02499 PCT/US95/07241 -49- J 6.9 Hz), 7.43-7.29 10 7.10 2 H, J 8.4 Hz), 6.90 2 H, J 8.7 Hz), 6.78 1 H, J 9.0 Hz), 5.56 (bs, 1 5.04 2 4.55 1 H, J 12.0 Hz), 4.47 1 H, J 12.0 Hz), 4.33-4.18 2 3.92-3.83 1 3.60 (dd, 1 H, J 9.6, 3.6 Hz), 3.45 (dd, 1 H, J 9.6, 3.6 Hz), 3.16 1 H, J 3.9 Hz), 3.03-3.00 (m, 2 2.18-1.93 2 1.44 (bs, 2 0.94 6 H, J 6.9 Hz), 0.85 3 H, J 6.9 Hz), 0.53 3 H, J 6.9 Hz); 19 F NMR (CDC1 3 6 -110.67 (dd, J 259, 5 Hz), 121.74 (dd, J 258, 18 Hz); mass spectrum, m/z 654 29), 626 (M 325, 92, 91 (100), 72. Anal. Calcd for
C
35
H
45
F
2
N
3 0 5 C, 67.18; H, 7.25; N, 6.72. Found: C, 67.34; H, 7.21; N, 6.66.
Step E Preparation of [1 4(S)]-2,4,5-Trideoxy-2,2difluoro-4-[[3-methyl-2-[[3-(4-morpholinyl)-1,3dioxopropyl]amino]-l-oxobutyl]amino]-N-[2-methyl-l- [(phenylmethoxy)methyl]propyl]-5-[4-(phenylmethoxy)phenyl]-L-glycero-pentanamide.
Reaction Scheme D, step The (R)-amino alcohol prepared above (169 mg, 0.270 mmol) is coupled under standard coupling conditions described previously, such as example 2, step A, with 2-(4-morpholinylcarbonyl)ethanoic acid (49 mg, 0.28 mmol, compound prepared in example to provide, after flash chromatography CH30H/CHC13), 185 mg of title compound as a tan powder: mp 87-94 0
C;
IR (KBr) Vmax 3428, 3325, 1651, 1535, 1512, 1454, 1238, 1115 cm- 1 19 F NMR (CDC1 3 6 -116.11 J 259 Hz), 119.49 (dd, J 259, 15 Hz); mass spectrum, m/z 809 (M 29), 781 (M 1) 509, 255, 243, 227, 88 (100).
Step F Preparation of final title compound.
Reaction Scheme D, step The above prepared alcohol (177 mg, 0.283 mmol) is oxidized under Swern conditions in a manner analogous to that described in WO 96/02499 PCT/US95/07241 example 1, step G to provide 170 mg (100%)of the final title compound after flash chromatography CH30H/CHC1 3 Recrystallization from CH2Cl 2 /ether provides 49 mg of final title compound as pale tan powder: mp 119-128 0 C; IR (KBr) vmax 3306, 1661, 1638, 1539, 1514, 1454, 1238, 1117 cm-1; 1 H NMR (CDCl 3 6 7.77 and 7.75 and 7.61 and 7.47 (4d, 1 H total, J 7.1, 7.1, 8.7, 8.9 Hz respectively), 7.43- 7.29 10 7.16-7.03 3H), 6.91-6.72 and 6.38-6.35 (2m, 3 H total), 5.74 and 5.53-5.46 and 5.24-5.17 (bs and 2m, 1 H total) 5.02 2 4.56 and 4.48 (2d, 1 H total, J 12.1 Hz), 4.55 and 4.47 (2d, 1 H total, J 12.0 Hz), 4.27 and 4.23 and 4.12 (3dd, 1 H total, J 8.8, 5.3 and 8.7, 5.7 and 7.3, 5.1 Hz respectively), 3.89-3.80 1 H), 3.69-3.59 7 3.52-3.40 3 3.32-3.21 3 2.94-2.75 1 2.29-1.87 2 0.94 and 0.93 and 0.92 and 0.91 and 0.89 and 0.88 and 0.84 and 0.81 and 0.80 and 0.76 and 0.59 and 0.55 (12d, 12 H total, J 6.8 and 7.0 and 7.0 and 6.9 and 6.9 and 7.0 and 7.0 and 7.0 and 6.8 and 6.8 and 6.9 and 6.8 Hz respectively); 19 F NMR (CDC1 3 6 -114.46 J 274 Hz), -112.74 -113.47 (d, J 274 Hz), Hydrate: -114.43 J 253 Hz), -116.47 (d, J 254 Hz), -120.63 J 253 Hz), -122.73 J 253 Hz); mass spectrum, m/z 807(M 29), 779 (M 401, 361, 243 (100), 227; exact mass calcd for C 42
H
53
F
2
N
4 0 8 779.3831, found 779.3878.
WO 96/02499 PCT/US95/07241 -51- Example Preparation of 4-Morpholine acetic acid, trifluoroacetic acid salt and 2 4 -Morpholinylcarbonyl)ethanoic acid i
S(B)I.
0 0 0 N-CH 2
OH.CF
3
CO
2 H II II O N-C C-OH 1A
B
Preparation of 4-Morpholine acetic acid, 1,1-dimethylethyl ester.
To a stirred solution of t-butyl bromoacetate (1.61 mL, 10.0 mmol) in tetrahydrofuran (25 mL) is added morpholine (1.74 mL, 20.0 mmol). The suspension is stirred for hours and then concentrated under vacuum. The residue is dissolved in methylene chloride (50 mL) with saturated sodium carbonate (50 mL). The layers are separated and the aqueous is extracted methylene chloride (2 x 25 mL). The organic extracts are combined, washed with saturated sodium carbonate (20 mL), brine (30 mL), dried over anhydrous magnesium sulfate, filtered and concentrated under vacuum.
The residue is triturated with ethyl acetate (15 mL), filtered and the filtrate concentrated under vacuum to provide the title compound (2.01 g) as a colorless oil; TLC Rf 0.45 (ethyl acetate, silica gel).
Preparation of final title compound Trifluoroacetic acid (15 mL) is added to the above prepared ester (1.00 g, 4.97 mmol). The solution is stirred for 5 hours and then concentrated under vacuum to provide a yellow oil. This is triturated with diethyl ether (25 mL) to provide the final title compound (1.06 g, 82%) as an off white solid; mp 118-121 0
C.
WO 96/02499 PCT/US95/07241 -52- Preparation of 2-(4-Morpholinylcarbonyl)ethanoic acid, methyl ester.
To a solution of methylmalonyl chloride (10.0 g, 73.2 mmol) in methylene chloride (200 mL) at 0°C is added rapidly dropwise a solution of morpholine (16.0 g, 0.183 mmol, 16.0 mL) in methylene chloride (50 mL). The reaction is stirred for 4 hours at room temperature. The reaction is then filtered and the filtrate is diluted with additional methylene chloride (200 mL). This is then washed with 1 N HC1, saturated sodium bicarbonate and brine. The organic is then concentrated under vacuum to provide a yellow oil, which is purified by flash chromatography (ethyl acetate, silica gel) to provide the title compound (9.7 g, 71%) as a pale yellow oil; Rf 0.28 (ethyl acetate).
Preparation of final title compound To a solution of the above prepared amide (1.70 g, 9.08 mmol) in methanol (45 mL) is added IN lithium hydroxide mL, 9.99 mmol). The reaction is stirred at room temperature for 2.5 hours. The pH is adjusted to 3 with IN HCL and the reaction is concentrated under vacuum. The residue is recrystallized from acetonitrile to provide the title compound (0.216 g, 14%) as a white solid.
WO 96/02499 PCT/US95/07241 -53- Example 6 [lR-(R*,2S*)-a,a-Difluoro-y-[2-(R)- [[(hydroxy)phenylacetyll-amino-3-methy-l-oxobutyllaminol- N-[2-methyl-l-[(phenylmethoxy)methyl]propyl]-B-oxo-4- (phenylmethoxy)-berzenepentanamide.
OBn O H 0 NH C FJH YNH OBn 0 0 Step A Preparation of [1 4(S)]-2,4,5-Trideoxy-4-[[2-[[4- [[2-[[[[(1,1-dimethylethyl)dimethylsilyl]oxylphenylacetyl]amino]-3-methyl-loxobutyl]amino]-2,2-difluoro-N-[2-methyl-i- [(phenylmethoxy)-methyl]propyll-5-[4-(phenylmethoxy)phenyl]-L-glycero-pentanamide.
Reaction Scheme step [1 4(S)1-4- Amino-2 4,5-trideoxy-2,2-difluoro-N-[ 2-methyl-l- [(phenylmethoxy)methyl]propyl-5-[4-(phenylmethoxy)phenyll-L-glycero-pentonamide (0.87 g, 1.6 mmol) prepared in example 5, step B, is coupled under standard coupling conditions in a manner analogous to that described previously, such as example 2, step A, with (R)-O-tertbutyldimethyl-silylmandelic acid (0.44 g, 1.2 mmol) prepared in example 1 to provide the title compound, after flash chromatography then 1:1 hexane/ethyl acetate), 0.36 g as a yellow oil: IR (film) Vmax 3405, 3325, 2959, 2932, 2895, 2876, 2861, 1684, 1657, 1512, 1470, 1454, 1244, 1221, 1179, 1098, 864 cm-1; 19 F NMR (CDCl 3 6 -117.20 J 258 Hz), -118.99 -120.55 J 262 Hz); mass spectrum (CI, 70 eV), m/z 874 509, 221 (100); exact mass calcd for C 4 9
H
6 6
F
2
N
3
Q
7 Si 874.4638, found 874.4651.
WO 96/02499 PCT/US95/07241 -54- Step B Preparation of final title compound.
Reaction Scheme steps and The above prepared alcohol(345 mg, 0.39 mmol) is oxidized under Swern conditions in a manner analogous to that described previously in example 1 to provide after flash chromatography (3:2 hexane/ethyl acetate), 154 mg 45%) of the ketone as a yellow glass: IR (film) Vmax 3408, 3298, 2961, 2932, 2861, 1655, 1613, 1514, 1470, 1454, 1246, 1179, 1098, 1072, 839 cm- 1 19 F NMR (CDCl 3 6 -112.68 -117.29 J 253Hz), -120.03 J 253 Hz); mass spectrum (CI, 70 eV), m/z 872 221 (100), 197; exact mass calcd for C 49
H
64
F
2
N
3 0 7 Si 872.4482, found 872.4500.
To a stirred solution of ketone in CH 2 C12 (4 mL) is added TFA/H 2 0 1 mL). The reaction mixture is allowed to stir at room temperature for 7 hours and then concentrated under vacuum. The residue is dissolved in
CH
2 C1 2 and the solution is washed with saturated aqueous sodium bicarbonate, brine, dried over anhydrous magnesium sulfate, filtered and concentrate under vacuum. The residue is purified by flash chromatography (1:1 hexane/ethyl acetate) to provide 85 mg of the final title compound as a white powder: IR (film) vmax 3397, 3314, 2965, 2934, 2874, 1750, 1688, 1663, 1532, 1514, 1468, 1454, 1242, 1179, 1113, 826 cm- 1 19 F NMR (CDC1 3 6 -112.30 J 271 Hz), -113.50 J 271 Hz), -117.54 J 254 Hz), -119.32 J 254 Hz); mass spectrum (CI, 70 eV), m/z 758 (M 107 (100), 91. Anal. Calcd for
C
43
H
49
F
2
N
3 07*0.6H 2 0: C, 67.19; H, 6.58; N, 5.47. Found: C, 67.11; H, 6.55; N, 5.39.
WO 96/02499 PCT/US95/07241 Example 7 Preparation of N-[l-[[[3,3-Difluoro-4-[[2-methyl-l- [(phenylmethoxy)-methyll-propyl]amino]-2,4-dioxo-l-[[4- (phenylmethoxy)phenyl]- methyll-butyl]amino]carbonyl]-2methylpropyl]-6-(4-morpholinyl-carbonyl)-3pyridinecarboxamide.
o j O O 0 NH H CFyNH 0 000 Step A Preparation of 6-(4-Morpholinylcarbonyl)-3pyridinecarboxylic Acid.
To a stirred 1 M solution of O-tert-butyl-N,N dicyclohexylisourea [see Mathias, Synthesis, 570, 1979, for preparation] in CH 2 C1 2 (20 mL) at 5 0 C under nitrogen is added 1.49 g (8.24 mmol) of 2-methyl isocinchomeronate [see Isagawa, et al., Nippon Kagaku Zasshi, 88, 553, 1967, for preparation] in portions over 2 minutes. After minutes, the cold water bath is removed. As dicyclohexylurea begins to precipitate, the reaction mixture is again be placed in a cool water bath at 15-20 0 C. The mixture is diluted with CH 2 C12 (5 mL) and allowed to stir at room temperature overnight. The mixture is then diluted with CH 2 C1 2 (30 mL), filtered, and the bluegreen solids are washed with CH 2 C1 2 The filtrate and washings are diluted with ether and washed with dilute aqueous sodium bicarbonate, water, brine, dried over anhydrous magnesium sulfate, filtered and concentrated under vacuum to provide 1.36 g of light blue solids. Flash chromatography (55:45 cyclohexane/ethyl acetate) provides 1.01 g of the diester below WO 96/02499 PCT/US95/07241 -56- 0
N
0 as white crystals. Recrystallization from ether/pentane provides the above diester as matted white needles: mp 111- 1120C; IR (KBr) Vmax 3420, 2984, 1711, 1379, 1310, 1290, 1246, 1134, 1126, 746 cm-l; 1H NMR (CDC13) 6 9.26 1 H, J 2.1 Hz), 8.39 (dd, 1 H, J 8.1, 2.1 Hz), 8.19 (dd, 1 H, J 8.1, 0.7 Hz), 4.04 3 1.63 9H); mass spectrum m/z 238 237 182, 179, 164, 57, (100); CI m/z 238 210, 182 (100). Anal. Calcd for C 12
H
15 N0 4 C, 60.75; H, 6.37; N, 5.90. Found: C, 60.80; H, 6.31; N, 5.75. A solution of the above diester (935 mg, 3.94 mmol) and morpholine (2.0 mL, 23 mmol) in THF (6 mL) are stirred at reflux under nitrogen for 24 hours.
Additional morpholine (2.0 mL, 23 mmol) is added and heating continued. After 3 more days, the solution is concentrated under vacuum and the residue is dissolved in ethyl acetate and the solution is washed twice with water.
The organic layer is concentrated under vacuum to provide 1.01 g of light yellow solid. Two recrystallizations from ether/pentane, with filtration through filter aid, provide 542 mg of amide ester below 0 0 0 O N N 0 as fine cream-colored crystals: mp 91-93 0 C; IR (KBr) Vmax 2984, 2965, 1707, 1634, 1370, 1317, 1287, 1169, 1132, 1117 cm-1 1 H NMR (CDC1 3 6 9.13 (dd, 1 H, J 2.1, 1.0 Hz), 8.36 (dd, 1 H, J 8.1, 2.1 Hz), 7.75 (dd, 1 H, J 8.1, 0.9 Hz), 3.83 4 3.73-3.55 4 1.62 9 H); mass spectrum, m/z 293 292, 123, 86 (100). Anal.
Calcd for C 1 5
H
20
N
2 0 4 C, 61.63; H, 6.90; N, 9.58. Found: C, WO 96/02499 PCT/US95/07241 -57- 61.62; H, 6.91; N, 9.64. HC1 gas is bubbled through a solution of the above amide ester (103.5 mg, 0.354 mmol) in
CH
3 NO2 (4-5 mL) for 20-25 minutes. After standing for an additional 20 minutes, the solution is concentrated under vacuum and the residue is triturated with acetone to provide the title compound (acid) as a pale yellow solid.
The material is combined with crude acid from a similar experiment (from 502 mg 14) and recrystallized from acetone to provide (335 mg of title compound as short, thick white needles: mp 181-183 0 C; IR (KBr) Vmax 2928, 2872, 1717, 1601, 1285, 1262, 1111 cm- 1 1 H NMR (CD30D) 6 9.26 1 H, J 1.1 Hz), 8.78 (dd, 1 H, J 8.1, 2.0 Hz), 1 H, J 8.1, Hz), 3.80 4 3.67 (nm, 2 3.51 (nm, 2 mass spectrum, m/z 277 (M 41), 265 (M 29), 238, 237 (M 1, 100). Anal. Calcd for C 11
H
12
N
2 0 4
C,
55.93; H, 5.13; N, 11.86. Found: C, 56.15; H, 5.32; N, 11.46.
Step B Preparation of [1 4(S)]-2,4,5-Trideoxy-2,2difluoro-4-[[3-methyl-2-[[[6-(4-morpholinylcarbonyl)-3pyridinyl]carbonyl]amino]-1-oxobutyl]-amino]-N-[2-methyl-l- [(phenylmethoxy)methyl]-propyl]-5-[4- (phenylmethoxy)phenyl]-L-glycero-pentanamide.
Reaction Scheme D, step The (S)-amino alcohol (185 mg, 0.296 mmol) prepared in example 5, step D, is coupled with the above prepared acid (77 mg, 0.33 mmol) under standard coupling conditions in a manner analogous to that described previously, such as example 2, step A, to provide after flash chromatography CH 3 0H/CHC13), 227 mg of the title compound as a white powder: mp 218-220 0
C;
IR (KBr) Vmax 3287, 1674, 1663, 1636, 1557, 1539, 1514, 1244, 1115 cm' 1 1 H NMR (CDC1 3 DMSO-d 6 6 9.05 1 H, J 1.8 Hz), 8.29 (dd, 1 H, J 8.1, 2.1 Hz), 8.03 1 H, J 9.3 Hz) 7.70 1 H, J 8.4 Hz), 7.46 1 H, J 8.4 Hz), 7.37-7.22 11 7.09 2 H, J 8.1 Hz), 6.67 2 H, J 8.4 Hz), 5.89-5.85 1 4.88 1 H, J WO 96/02499 PCT/US95/07241 -58- 11.7 Hz), 4.83(d, 1 H, J 11.7 Hz), 4.54 1 H, J 12.0 Hz), 4.47 1 H, J 12.0 Hz), 4.45-4.35 1 H), 4.31 1 H, J 9.0 Hz), 4.24-4.13 1 3.92-3.82 1 3.77 4 3.64-3.48 6 3.08-3.02 (m, 1 2.76 (dd, 1 H, J 14.4, 10.5 Hz), 2.15-1.92 2 0.94 6 H, J 6.9 Hz), 0.88 6 H, J 6.6 Hz); 19 F NMR (CDC1 3 DMSO-d 6 6 -109.78 J 256 Hz), -122.05 (dd, J 256, 19 Hz); mass spectrum, m/z 872 (M 29), 844 509 (100), 418, 318, 290, 219. Anal. Calcd for
C
46
H
55 F2N508: C, 65.47; H, 6.57; N, 8.30. Found: C, 65.07; H, 6.65; N, 9.12.
Step C Preparation of final title compound.
Reaction Scheme D, step The above prepared alcohol (196 mg, 0.232 mmol) is oxidized under under Swern conditions in a manner analogous to that described previously in example 1, to provide 196 mg (100%) of crude final title compound as an orange oil after flash chromatography
CH
3 0H/CHC13). Recrystallization from
CH
2 Cl 2 /pentane, then CH 2 C12/ ether, provides 100 mg of pure final title compound as cream granules: mp 108-135 0
C;
IR (KBr) vmax 3422, 3306, 1638, 1535, 1514, 1454, 1115 cm- 1 1 H NMR (CDC13) 6 8.95-8.87 1 8.16 and 8.13 and 8.12-8.08 (2dd and m, 1 H total, J 2.2, 1.1 Hz), 7.74 and 7.71 end 7.68-7.67 (2dd and m, 1 H total, J 5.8, 0.7 Hz), 7.43-7.27 10 7.14-7.04 and 7.13 and 7.09 and 7.06 (m and 3d, 2 H total, J 8.5, 8.8, 8.8 Hz respectively), 6.95-6.77 and 6.88 and 6.85 (m and 2d, 4H total, J 8.7 Hz), 6.49 and 6.39 and 6.23 (3d, 1 H total, J 7.2, 6.7, Hz respectively), 5.60-5.56 and 5.46 and 5.33-5.21 (m and bs and m, 1 H total), 5.02 and 4.98 and 4.93 (3s, 2 H total) 4.57-4.40 3H), 3.81 (bs, 5 3.67-3.59 3.50-3.43 1H), 3.36-3.27 1 2.95-2.76 1 2.17-1.91 2 0.96-0.88 and 0.83 and 0.82 and 0.71 and 0.59 (m and 4d, 12 H total, J 6.8 Hz); 19F NMR (CDC13) d -111.54 J 270 Hz), -112.47 J 269 WO 96/02499 PCT/US95/07241 -59- Hz), -113.76 J 270 Hz), -114.62 J 270 Hz), Hydrate: -114.80 J 253 Hz), -118.53 -118.68 -122.82 J 254 Hz); mass spectrum, m/z 870 (M* 29), 842 (M 197, 194, 91 (100); exact mass calcd for C 46
H
54
F
2
N
5 0s 842.3940, found 842.3994. Anal. Calcd for
C
46
H
53
F
2
N
5 0s: C, 65.62; H, 6.35; N, 8.32. Found: C, 64.80; H, 6.46; N, 8.27.
Example 8 Preparation of 7,7-Difluoro-4,12-bis(l-methylethyl)-6,8,11trioxo-l-phenyl-9-[[4-(phenylmethoxy)phenyllmethyll-2-oxa- 5,10,13-triazatetradecan-14-oic Acid, 3-Pyridinylmethyl Ester.
OBn o 0 NH CF Bn 0 0 Step A Preparation of 3 -Pyridinylmethoxy)carbonyl]-L-valine.
To a stirred suspension of L-valine methyl ester hydrochloride (2.0 g, 12 mmol) in toluene (15 mL) is added triphosgene (1.78 g, 6 mmol) and DMF (0.15 mL, 2 mmol).
The heterogeneous mixture is allowed to stir at room temperature for 30 minutes. To this mixture is added tributylamine (0.1 mL, 0.4 mmol) dropwise over 2 minutes.
The reaction mixture is heated to reflux for 1 hour.
Analysis by IR spectroscopy shows a strong signal at 2250 cm 1 indicating the presence of isocyanate. The now homogeneous solution is cooled to o0C and 3-pyridylcarbinol (0.9 mL, 9 mmol) is added dropwise over 10 minutes. A white precipitate forms during the addition. The reaction mixture is warmed to room temperature and is allowed to stir for 16 hours. Additional 3-pyridylcarbinol (1.8 mL, WO 96/02499 PCT/US95/07241 18 mmol) is added and the cloudy mixture is heated to reflux for 3 hours. The solution is concentrated under vacuum, diluted with ethyl acetate and washed with three portions of water. The solution dried over anhydrous magnesium sulfate, filtered and concentrated under vacuum to provide an oil. Flash chromatography (8:1 CH 2 Cl 2
/C
2
H
5
OH)
of the oil provides 1.46 g of pyridinylmethoxy)carbonyl]-L-valine, methyl ester as a yellow oil: 1 H NMR (CDC13) 6 8.61 1 8.56 1 H, J 6.5 Hz), 7.71 1 H, J 10.2 Hz), 7.29 (dd, 1 H, J 8.1, 6.6 Hz), 5.38 (bs, 1 5.13 2 4.29 (dd, 1 H, J 8.2, 5.9 Hz), 2.12 1 0.92 3 H, J 6.3 Hz), 0.88 3 H, J 6.3 Hz); [a]20D +9.250 (c 0.85, CHC1 3 Anal. Calcd for C 13
H
18
N
2 0 4 C, 58.65; H, 6.77; N, 10.53.
Found: C, 57.62; H, 7.03; N, 10.47.
Alternatively the N-[(3-pyridinylmethoxy)carbonyl]-Lvaline, methyl ester can be prepared as follows: L-valine methyl ester (4.25 g, 34 mmol, freshly prepared from the hydrochloride salt by neutralization with 50% aqueous NaOH) is added to a stirred mixture of 1,1 '-carbonyldiimidazole (4.86 g, 30 mmol) in CH 2 C1 2 (30 mL) over 15 minutes. After more minutes, 3-pyridylcarbinol (5.0 mL, 51 mmol) is added dropwise to the homogeneous solution. The resulting solution is heated at 45 0 C for 3 hours and then allowed to stir at room temperature overnight. The CH 2 C12 is removed under vacuum, and the residue is dissolved in toluene (70 mL). The solution is heated at 70 0 C for 6 hours, and then concentrated under vacuum. The residue is dissolved in CH 2 C1 2 and the resulting solution is washed three times with water, dried over anhydrous magnesium sulfate, filtered and concentrate to provide an oil, which is purified by flash chromatography to provide 4.6 g (51%) of the N-[(3-pyridinylmethoxy)carbonyl]-L-valine, methyl ester as a yellow oil.
WO 96/02499 PCT/US95/07241 -61- To a stirred solution of the pyridinylmethoxy)carbonyl]-L-valine, methyl ester (1.4 g, 5.3 mmol) in CH 3 OH (6 mL) is added LiOH.H 2 0 (0.24 g, 5.8 mmol). The heterogeneous mixture is allowed to stir at room temperature for 16 hours. The reaction mixture is diluted with water until completely homogeneous, acidified with 0.5 N HC1 (11.6 mL, 5.8 mmol) and concentrated under vacuum to provide a white solid, which is crystallized from 8:1 CH 2 C1 2
/C
2 H50H to the title compound 0.53 g as a white crystalline solid: mp 242-244 0 C; 1 H NMR (DMSO-d 6 6 8.9 2 8.5 1 H, J 8 Hz), 8.05 1 7.7 1 H, J 9 Hz), 5.22 2 3.9 1 2.05 1 0.85 6 H).
Step B Preparation of [1 3 4(S)]-2,4,5-Trideoxy-2,2difluoro-4-[[3-methyl-l-oxo-2-[[(3pyridinylmethoxy)carbonyl]amino]butyl]amino]-N-[2-methyl-l- [(phenylmethoxy)methyl]propyl]-5-[4-(phenylmethoxy)phenyl]-L-qlycero-pentanamide.
The amino alcohol prepared in example 5, step D (0.28 g, 0.53 mmol) is coupled with the above prepared acid (0.19 g, 0.74 mmol) under standard coupling conditions described previously, such as example 2, step A to provide, after crystallization from ethyl acetate, 0.20 g of title compound as a white powder: mp 187-191C; IR (KBr) Vmax 3426, 3300, 2960, 1698, 1674, 1657, 1541, 1512, 1246, 1177, 1151, 1105, 1028, 698 cm-l; 1 H NMR (DMSO-d 6 6 8.58 1 H, J 1.6 Hz), 8.49 (dd, 1 H, J 4.7, 1.4 Hz), 7.76 1 H, J 7.9 Hz), 7.46-7.24 13 7.13-7.07 (2d, 2 H total, J 8.8 Hz), 6.88 and 6.81 (2d, 2 H total, J 8.8, 8.3 Hz), 8.34, 7.97, and 6.18 (3d, 1 H total), 5.08 and 5.03 (d and s, 2 H total, J 5.7 Hz), 4.99 2 H, J 2.2 Hz), 4.46 2 H, J 3.8 Hz), 4.21 1 4.06-3.92 1 3.87-3.73 2 3.52 2 H, J 6.9 Hz), 2.93 (d, 1 H, J 12.2 Hz), 2.6 1 1.93-1.75 2 0.87 (apparent t, 8 H, J 6.6 Hz), 0.68 (dd, 4 H, J 6.8, 6.6 WO 96/02499 PCT/US95/07241 -62- Hz); 19 F NMR (DMSO-d 6 6 major diastereomer: -108.9 (dd, J 251, 7 Hz), -119.8 (dd, J 245, 19 Hz), minor diastereomer: -111.6 (dd, J 252, 8 Hz), -117.1 (dd, J 250, 18 Hz); mass spectrum, m/z 761 (M 555, 138, 121, 110, 92. Anal. Calcd for C 42
H
50
F
2
N
4 0 7 C, 66.32; H, 6.58; N, 7.37. Found: C, 62.43; H, 6.36; N, 6.79.
Step C Preparation of final title compound.
Reaction Scheme D, step In a manner analogous to that described previously in example 1, the above prepared alcohol is oxidized under Swern conditions to provide after flash chromatography (8:1 CH 2 C1 2
/C
2
H
5 0H), 100 mg of final title compound as a white powder: mp 108-111 0 C; IR (KBr) Vmax 3416, 3308, 2963, 1696, 1660, 1537, 1514, 1246, 1115, 1028 cm-1; 1 H NMR (DMSO-d 6 6 8.91 1 H, J 8.8 Hz), 8.58-8.51 2 7.75 1 H, J 7.7 Hz), 7.45- 7.23 12 7.16 1 H, J 8.6 Hz), 6.90(d, 2 H, J 8.6 Hz), 5.06-4.99 5 4.45 2 H, J 3.1 Hz), 3.92-3.8 2 3.52 2 3.10 (dd, 1 H, J 14.9, 3.1 Hz), 2.69 (dd, 1 H, J 14.1, 9.9 Hz), 1.87 2 H), 0.84 and 0.78 (2dd, 10 H total, J 10.8, 7.1 Hz and J 6.4, 4.8 Hz respectively), 0.60 (dd, 2 H, J 8.7, 6.9 Hz); 13C NMR (DMSO-d 6 6 171.2 157.15, 155.8, 149.0, 148.95, 148.9, 138.3, 137.1, 135.5, 132.6, 130.2, 130.07, 128.8, 128.7, 128.37, 128.16, 127.75, 127.67, 127.6, 127.5, 127.44, 127.36, 123.43, 114.5, 71.9, 69.5, 69.45, 69.1, 69.0, 63.16, 59.6, 55.25, 54.76, 54.67, 33.8, 33.75, 30.35, 28.8, 28.77, 19.4, 19.33, 18.98, 18.59, 17.86; 19 F NMR (DMSO-d 6 6 -111.36 J 272 Hz), -114.36 J 270 Hz), -112.96 -117.06 J 255 Hz), -120.00 J 250 Hz); mass spectrum, m/z 759 460, 110, 91.
Anal. Calcd for C 42
H
48
F
2
N
4 0 7
*H
2 0: C, 64.94; H, 6.44; N, 7.22.
Found: C, 64.70; H, 6.36; N, 7.24.
WO 96/02.499 PCTIUS95/07241 -63- Example 9 Preparation of ]-c,a-Difluoro-y-[ [3-methyl-2- 4 -(4-morpholinylsulfonvl)benzovllamino.>..
oxobutyl ]amino] -N-[2-methyl-i- [(phenylmethoxv)methylpoy 1-oo4(hnlehx) benzenep~entanamide.
0 0 OBn H C Ff N H Bn Step
A
Preparation of 4 (S)1-2,4,5-Trideoxy-2,2.
difluoro-4-[ t3-methvl-2-[ morpholinylsulfonyl )benzoyl 11amino] -1-oxobutyl -amino] -N-l2methyl-l-[(phenvlmethoxy)mthylpropyl.5[4..(phenylmethoxy)phenyl ]-L-glycero-pentanamide.
Reaction Scheme D, step The (S)-amino alcohol prepared in example 5, step D (161 mg, 0.26 rnmol) is coupled with the acid (77 mg, 0.28 mmol) of the following structure 0 C0 2
H
S
11 0 [prepared as described by Sycheva, T.P. et al., Sbornik Statei Obshchei him, Akad. Nauk 1, 568-571 (1953)] under standard coupling conditions in a manner analogous to that described previously, such as example 2, step A, to provide after flash chromatography (1:1 ethyl acetate/CH 2 Cl 2 140 mg of the title compound as a white solid. Recrystallization from ethyl acetate/cyclohexane provides the title compound as a white powder: mp 172.5-190.5'C (172.5-174, then 189.5-190-5 0
IR
WO 96/02499 PCT/US95/07241 -64- (KBr) Vmax 3420, 3322, 2965, 1643, 1534, 1514, 1171, 1113 cm- 1 1 H NMR (CDC13) 6 7.90 2 H, J 8.4 Hz), 7.79 (d, 2 H, J 8.3 Hz), 7.4-7.25 10 7.08 2 H, J 8.6 Hz), 6.78 (d overlapping m, 4 H, J 8.5 Hz), 6.65 1 H, J 8.6 Hz), 4.93 2 4.53 1 H, J 12.0 Hz), 4.47 1 H, J 12.0 Hz), 4.45-4.24 4 3.82 1 3.73-3.65 (nm, 4 3.62 (dd, 1 H, J 9.7, 4.1 Hz), 3.46 (dd, 1 H, J 10.0, 3.9 Hz), 3.1-2.85 6 2.12 1 1.98 1 0.97-0.89 12 19 F NMR (CDC1 3 6 -110.18 (dd, J 261, 4 Hz), -121.09 (dd, J 261, 18 Hz); mass spectrum, m/z 879 (M 1, 100), 527, 325, 254. Anal. Calcd for C 46
H
56
F
2
N
4 09S: C, 62.85; H, 6.42; N, 6.37. Found: C, 62.65; H, 6.43; N, 6.26.
Step B Preparation of final title compound.
Reaction Scheme D, step In a manner analogous to that described in example 1 the above prepared alcohol (133 mg, 0.151 mmol) is oxidized under Swern conditions to provide 82 mg of a mixture of diastereomers of final title compound after flash chromatography (3:2 ethyl acetate/cyclohexane). The mixture partially crystallizes from cyclohexane/ethyl acetate upon evaporation of the solvent. The tacky pale yellow solid is triturated with ether to provide 59 mg of a single diastereomer of final title compound as a white powder: mp 114-116 0 C; IR (KBr) Vmax 3426, 2971, 1686, 1663, 1512, 1171, 1115 cm- 1 1H NMR (CDC13) 6 7.92 2 H, J 8.3 Hz), 7.81 2 H, J 8.3 Hz), 7.41-7.29 10 7.06 2 H, J 8.6 Hz), 6.85 2 H, J 8.6 Hz), 6.78 2 H, J 8.6 Hz), 6.29 1 H, J 6.7 Hz), 5.26 1 4.99 2 H, major peak), 4.55 1 H, J 12.0 Hz), 4.48 1 H, J 12.0 Hz), 4.43 (dd, 1 H, J 8.4, 6.4 Hz), 3.84 1 3.72 (narrow m, 4 3.63 (dd, 1 H, J 9.8, 3.8 Hz), 3.47 (dd, 1 H, J 9.7, 3.8 Hz), 3.31 (dd, 1 H, J 14.4, 5.0 Hz), 3.00-2.87 5 2.14 1 2.00 1 0.96 (d, 3 H, J 6.7 Hz), 0.95 3 H, J 6.7 Hz), 0.94 3 H, WO 96/02499 PCT/US95/07241 J 6.7 Hz), 0.92 3 H, J 6.7 Hz); 1 9 F NMR (CDC13) d 112.45 J 270 Hz), -113.69 J 270 Hz),-117.94 (d, J 255 Hz), -119.29 J 255 Hz); mass spectrum (CI), m/z 905 29), 877 (M 525, 507, 401, 353 (100), 197, 107. Anal. Calcd for C 46
H
54
F
2
N
4 0 9 S.0.5 H 2 0: C, 62.36; H, 6.26; N, 6.32. Found: C, 62.24; H, 6.30; N, 6.27.
Example Preparation of [lR-(lR*,2S*)]-a,a-Difluoro-y-[[3-methyl-2- [[4-(4-morpholinylcarbonyl)benzoyl]amino]-1oxobutyl]amino]-N-[2-methyl-l- [(phenylmethoxy)methyl]propyl]-B-oxo-4-(phenylmethoxy)benzenepentanamide.
0 r 0Bn
NH
0 NH CF H 0 o o Step A Preparation of [1 3C, 4(S)]-2,4,5-Trideoxy-2,2difluoro-4-[[3-methyl-2-[[4-(4morpholinylcarbonyl)benzoyl]amino]-1-oxobutyl]-amino]-N-[2methyl-l-[(phenylmethoxy)methyl]propyl]-5-[4-(phenylmethoxy)phenyl]-L-qlycero-pentanamide.
Reaction Scheme D, step The amino alcohol (171 mg, 0.274 mmol) prepared in example 5, step D is 3 coupled with 4 4 -morpholinylcarbonyl)benzoic acid (68 mg, 0.29 mmol, prepared in example 10a) under standard coupling conditions, such as example 2, step A, in a manner analogous to that described previously to provide, after flash chromatography CH30H/CHC13), 217 mg of the title compound as a yellow powder: IR (KBr) vmax 3428, 3322, 1636, 1534, 1512, 1456, 1279, 1258, 1244, 1115 cm- 1 19 F NMR (CDC1 3 6 -110.19 J 259 Hz), -117.26 J 259 Hz), -120.14 J 259 Hz), -121.40 (dd, J 259, WO 96/02499 PCT/US95/07241 -66- Hz) impurities; mass spectrum, m/z 871 (M 29), 843 527, 509, 329, 317 (100), 289, 250, 218.
Step B Preparation of final title compound.
Reaction Scheme D, step The above prepared alcohol is oxidized under Swern conditions as follows: To a stirred solution of the above prepared alcohol (215 mg, 0.255 mmol) in anhydrous CH 2 C1 2 (3 mL) and anhydrous DMSO (0.36 mL, 5.1 mmol) at -45 0 C is added 2 M oxalyl chloride/CH 2 Cl 2 (1.0 mL) over 5 minutes. The solution is allowed to stir at -40 to -30 0 C for 2.5 hours. The solution is then cooled to -70 0 C and diisopropylethylamine (0.67 mL, 3.8 mmol) is added over 5 minutes. The solution is allowed to warm to room temperature, diluted with CH 2 C1 2 washed twice with water, dried over anhydrous magnesium sulfate, filtered and concentrated under vacuum. The residue is purified by flash chromatography CH 3 0H/CHC1 3 to provide 210 mg of final title compound as an amber oil.
Recrystallization from CH2Cl 2 /pentane provides 80 mg (37%) of final title compound as an amber powder: mp 87-92 0 C; IR (KBr) Vmax 3306, 1636, 1534, 1514, 1454, 1279, 1258, 1244, 1115 cm- 1 1 H NMR (CDC13) 6 7.82-7.75 2 7.48-7.30 12 7.11-7.04 2 6.91-6.82 2 6.78 (d, 1 H, J 8.7 Hz), 6.68(d, 1 H, J 8.7 Hz), 6.44 and 6.33 (2d, 1 H, J 6.9 Hz), 5.31-5.22 1 5.05-4.94 2 4.57-4.38 3 3.88-3.28 12 2.94-2.81 (m, 1 2.16-1.93 2 0.96-0.58 12 19F NMR (CDC1 3 6 -111.56 J 270 Hz), -112.56 J 270 Hz), -113.63 J 270 Hz), -114.54 J 270 Hz), Hydrate: -113.53 J 292 Hz), -114.85 J 252 Hz), -118.71 J 292 Hz), -122.88 J 252 Hz); mass spectrum, m/z 869 (M 29), 841 (M 525, 317, 279, 218, 197, 157 (100); exact mass calcd for C 47
H
55
F
2
N
4 0 8 841.3988, found 841.4043. Anal. Calcd for C 47
H
54
F
2
N
4 0 8 C, 67.13; H, 6.47; N, 6.66. Found: C, 65.80; H, 6.60; N, 6.69.
WO 96/02499 PCT/US95/07241 -67- Example Preparation of 4-(4-morpholinylcarbonyl)benzoic acid.
O N-C OH Preparation of 4-(4-morpholinylcarbonyl)benzoic acid methyl ester.
To a stirred suspension of monomethyl terephthalate (25.0 g, 0.14 mmol) in methylene chloride (500 mL) and dimethylformamide (4 mL) is added dropwise oxalyl chloride (12.1 mL, 0.14 mol) with vigorous gas evolution. After gas evolutions ceases, the reaction is stirred for 45 minutes and then cooled in an ice-water bath. Morpholine (48.4 mL, 0.56 mol) is then added resulting in an exotherm. After the addition is complete the reaction is allowed to warm to room temperature and stirred for 1 hour. The reaction mixture is then washed with 0.5 N HC1 (2 x 500 mL), halfsaturated sodium bicarbonate (2 x 500 mL), water (2 x 500 mL) and brine (300 mL). The reaction is dried over anhydrous magnesium sulfate, filtered and concentrated under vacuum to provide the title compound (33.8 g, 98%) as a white solid; mp 74-76 0
C.
Preparation of final title compound.
to a stirred solution of the above ester (50.1 g, 0.20 mmol) in methanol (800 mL) is added IN lithium hydroxide (241 mL, 0.24 mol) followed by water (160 mL). After 3 stirring at room temperature for 7 hours, additional water (100 mL) is added. The reaction is then stirred an additional 24 hours and then concentrated to remove the methanol. The aqueous is extracted with methylene chloride (2 x 100 mL) and diethyl ether (200 mL). The aqueous layer was then cooled in an ice-water bath and acidified to pH 1 with 12 N HC1 (ca. 20 mL). Filtration provides crude white solid. The solid material is suspended in methylene chloride (2 L) and heated to reflux. It is then cooled to WO 96/02499 PCT/US95/07241 -68room temperature, treated with anhydrous magnesium sulfate, and filtered. The filtrate is concentrated under vacuum to provide the final title compound (42.2 g, 89%) as a white solid; mp 194-196 0
C.
WO 96/02499 PCT/US95/07241 -69- Example 11 Preparation of N-[l-[[[3,3-Difluoro-4-[[2-methyl-l- [(phenylmethoxy)methyl]-propyl]amino]-2,4-dioxo-l-[[4- (phenylmethoxy)phenyl]-methyl]butyl]amino]carbonyl -2methylpropyl]-4-morpholineacetamide.
S.OBn 0 N NH
CF
n H I
F
I OBn O Step A Preparation of[l 4(S))-2,4,5-Trideoxy-2,2difluoro-4-[[3-methyl-2-[[(4-morpholinyl)acetyl]aminoloxobutyl]amino]-N-[2-methyl-l- [(phenylmethoxy)methyl]propyl]-5-[4-(phenylmethoxy)phenyl]- L-glycero-pentanamide.
Reaction Scheme D, step The (R,S)-amino alcohols (201 mg, 0.321 mmol) prepared in example 5, step D are coupled with 4-morpholine acetic acid, trifluoro acid salt (92 mg, 0.35 mmol, compound prepared in example under standard coupling conditions in a manner analogous to that described previously [with the addition of 1.1 eq. of NMM] to provide after flash chromatography CH 3 OH/CHCl 3 224 mg of title compound as a beige powder: mp 110- 117 0 C; IR (KBr) Vmax 3298, 2963, 1674, 1649, 1545, 1512, 1238, 1115, 698 cm-; 19 F NMR (CDC13) 6 major diastereomer- 3 110.56 (dd, J 260, 5 Hz), -120.79 (dd, J 260, 18 Hz); mass spectrum, m/z 781 29), 753 (M 1, 100), 752 (M 291, 100, 91.
Step B 3 Preparation of final title compound.
Reaction Scheme D, step In a manner analogous to that described in example 1 the above prepared alcohol (206 mg, 0.274 mmol) is oxidized under Swern conditions to WO 96/02499 PCT/US95/07241 provide 170 mg of an orange glass after flash chromatography CH30H/CHC1 3 Recrystallization from
CH
2 Cl 2 /ether provides 70 mg of final title compound as sticky amber crystals: mp 48-53 0 C; IR (CHC1 3 Vmax 2969, 1686, 1512, 1242, 1117, 760, 750, 731 cm-; 1 H NMR (CDC1 3 6 7.55-7.50 1 7.43-7.29 10 7.14-7.07 2 6.92-6.78 3 6.52-6.49 and 6.40-6.37 (2m, 1 H total), 5.33-5.19 1 5.03 (apparent d, J 3.6 Hz), 4.55 1 H, J 11.9 Hz), 4.48 and 4.47 (2d, 1 H total, J 11.9 Hz), 3.88-3.80 1 3.74-3.61 5 3.50- 3.44 1 3.32-3.25 1 2.99-2.81 3 H), 2.49-2.46 4 2.14-1.95 2 1.28-1.24 and 0.95- 0.70 and 0.50 (2m and d, 12 H total, J 6.7 Hz); 19 F NMR (CDC1 3 6 -111.32 J 273 Hz), -112.96 -114.64 (d, J 272 Hz); mass spectrum, m/z 779 (M 29), 751 (M 226, 100 (100), 86; exact mass calcd for C 41
H
53
F
2
N
4 0 7 751.3882, found 751.3892. Anal. Calcd for
C
4 1
H
5 2
F
2
N
4 0 7 .1.3H 2 0: C, 63.60; H, 6.77; N, 7.24. Found: C, 63.64; H, 6.80; N, 6.84.
Example 12 Preparation of N-[l-[[[3,3-Difluoro-4-[[2-methyl-l- [(phenylmethoxy)methyl]-propyl]amino]-2,4-dioxo-l-[[4- (phenylmethoxy)phenyl]methyl]-butyl]amino]carbonyl]-2methylpropyl]-1H-imidazole-l-acetamide.
H OBn 0 H II IIHII
*HCI
0 0 0 °X Step A Preparation of 1H-Imidazole-l-acetic Acid*HCl.
To a stirred solution of imidazole (1.50 g, 22.0 mmol) in CH 2 C1 2 (25 mL) at 0°C under nitrogen is added dropwise tert-butyl bromoacetate (1.62 mL, 10.0 mmol). The ice bath WO 96/02499 PCT/US95/07241 -71is removed and the solution is allowed to stir at room temperature for 22 hours. The solution is concentrated under vacuum and the residue is partitioned between ethyl acetate/ water. The organic layer is washed with water and concentrated under vacuum to provide 1.22 g of tertbutyl 1H-imidazole-l-acetate as a white crystalline solid: mp 110-113 0 C; 1 H NMR (CDC1 3 6 7.49 1 6.95 1 4.59 2 1.47 9 HC1 gas was bubbled through a solution of the tert-butyl 1H-imidazole-l-acetate in CH 3
NO
2 (4-5 mL) for 20-25 minutes. After standing for minutes more, the solution is concentrated under vacuum and recrystallized from CH 3
CN/CH
3 0H to provide the title compound in 70% yield: mp 206-209 0 C; 1 H NMR (DMSO-d 6 6 9.12(d, 1 H, J 1.4 Hz), 7.73 (narrow m, 1 7.68 1 H, J 1.4 Hz), 5.15 2 Anal. Calcd for
C
5
H
6
N
2 0 2 .HC1: C, 36.94; H, 4.34; N, 17.23. Found: C, 37.21; H, 4.36; N, 17.39.
Step B Preparation of[1 4(S)]-2,4,5-Trideoxy-2,2difluoro-4-[[2-[(lH-imidazol-l-ylacetyl)amino]-3-methyl-loxobutyl]amino]-N-[2-methyl-l- [(phenylmethoxy)methyl]propyl]-5-[4-(phenylmethoxy)phenyl]- L-qlycero-pentanamide.
Reaction Scheme D, step To a solution of the amino alcohol (205 mg, 0.328 mmol) prepared in example step D in anhydrous CH 2 C12 (4 mL) and anhydrous DMF (7 mL) is added 1H-imidazole-l-acetic acid*HCl(56 mg, 0.34 mmol) prepared above and NMM (35 iL, 0.32 mmol). Not all of the 1H-imidazole-l-acetic acid dissolves. HOBT (53 mg, 0.34 mmol) and EDC (66 mg, 0.34 mmol) are then added and the resulting mixture is allowed to stir overnight at room temperature. The mixture is concentrated under vacuum and the residue is dissolved in ethyl acetate. The organic layer is washed with water, 10% aqueous sodium bicarbonate, brine, dried over anhydrous magnesium sulfate, filtered and concentrated under vacuum to provide yellow solids which WO 96/02499 PCT/US95/07241 -72contain two major spots by TLC (10% CH30H/CHC1 3 The two compounds are separated by chromatography (the first is eluted with 4% CH 3 0H/CHCl 3 the second with 1:1
CH
3 0H/CHC1 3 The less polar material is the formamide of of the starting material while the more polar material is the title compound (110 mg, 45%) which is isolated as a white powder: mp 190-193 0 C; IR (KBr) Vmax 3430, 3297, 1678, 1647, 1549, 1512, 1240, 1082 cm- 1 19 F NMR (CDC1 3 DMSO-d6) 6 major diastereomer -115.66 (dd, J 259, 7 Hz), -120.48 (dd, J 259, 18 Hz); mass spectrum, m/z 762(M 29), 734 (M 91 (100).
Step C Preparation of final title compound.
Reaction Scheme D, step In a manner analogous to that described in example 1 the above prepared alcohol (101 mg, 0.138 mmol) is oxidized under Swern conditions to provide 101 mg (100%) of an orange oil after flash chromatography (2:1 CHC13/CH 3 0H). Recrystallization from
CH
2 C1 2 /pentane provides 50 mg of final title compound as a beige solid: mp 112-116 0 C; IR (KBr) vmax 3428, 3295, 2963, 1647, 1551, 1512, 1240, 1113, 698 cm- 1 1 H NMR (DMSOd 6 6 9.00 and 8.93 and 8.66 and 8.64 (4d, 1 H total, J 9.0, 7.2, 7.2 Hz respectively), 8.16 (apparent triplet, 1 H, J 8.4 Hz), 7.59 (bs, 1 7.46-7.26 11 7.15 and 7.14 (2d, 1 H total, J 8.6 Hz), 7.06 (bs, 1 6.90 and 6.89 (apparent 2d, 3 H, J 8.7 Hz), 5.05 (s, 2 5.01 (dd, 1 H, J 7.5, 3.4 Hz), 4.79-4.61 2 H), 4.50-4.39 2 4.30-4.20 1 3.85-3.76 1 H), 3.54-3.50 2 3.20-3.08 1 2.72-2.57 1 H), 2.00-1.72 2 1.27-1.21 and 0.87-0.69 and 0.61-0.52 12 H total); 19 F NMR (DMSO-d 6 6 -108.87 J 267 Hz), -109.82 J 268 Hz), -111.37 J 268 Hz), 112.32 J 254 Hz), 112.58 J 268 Hz), -117.61 (d, J 254 Hz); mass spectrum, m/z 760 (M 29), 732 (M 712, 180, 91 (100), 69; exact mass calcd for C 40
H
48
F
2
N
5 0 6 732.3573, found 732.3541. Anal. Calcd for C 40
H
47 F2N 5 06*2H20: WO 96/02499 PCT/US95/07241 -73- C, 62.57; H, 6.69; N, 9.12. Found: C, 62.07; H, 6.49; N, 9.05.
Example 13 Preparation of [l-[[[3,3-Difluoro-2,4-dioxo-l-[[4- (phenylmethoxy)phenyl]-methyl]-4-[(2pyridinylmethyl)amino]butyl]amino]carbonyl]- 2methylpropyl]-carbamic Acid, 3-Pyridinylrethyl Ester.
OBn 1 0 N ONH CF 2
NH
O O O Step A Preparation of [1 3&]-2,4,5-Trideoxy-4-[[(1,1dimethylethoxy)carbonyl]-amino]-2,2-difluoro-5-[4- (phenylmethoxy)-phenyll-N-(2-pyridinylmethyl)-L-qlyceropentanamide.
Reaction Scheme A, step To a stirred solution of 4 -tert-butoxycarbonylamino-2,2-difluoro-3-hydroxy-5-(4benzyloxy)phenylpentanoic acid, ethyl ester (1.5 g, 3.1 2 mmol) prepared in example 1 step B, in dry THF (16 mL) is added 2-(aminomethyl)pyridine (0.38 mL, 3.8 mmol). The homogeneous solution is heated at reflux for 4 hours and then allowed to cool to room temperature. A thick white precipitate forms upon cooling which is filtered and washed with ether to provide 0.80 g of title compound as a white powder: mp 166-170°C; 1 H NMR (DMSO-d 6 6 9.23 (bs, 1 8.51 1 7.78 1 H, J 7.5 Hz), 7.5-7.25 (m, 8 7.12 2 H, J 8.4 Hz), 6.91 2 H, J 8.4 Hz), 6.69 1 H, J 10.4 Hz), 6.19 1 H, J 8.9 Hz), 5.08 2 4.58-4.32 2 4.15-3.91 1 H), 3.82 1 2.95 1 H, J 10.9 Hz), 1.15 9 H); 19 F NMR (DMSO-d 6 6 major diastereomer: -103.30 (dd, J 253, 9 Hz), -117.67 (dd, J 252, 17 Hz), minor WO 96/02499 PCT/US95/07241 -74diastereomer: -111.37 (dd, J 254, 8 Hz), -121.27 (dd, J 256, 18 Hz).
Step B Preparation of 4(S)]-2,4,5-Trideoxy-2,2-difluoro-4- [[3-methyl-l-oxo-2-[[(3pyridinylmethoxy)carbonyl]amino]butyl]amino]-5-[4- (phenylmethoxy)phenyl]-N-(2-pyridinylmethyl)-L-qlyceropentanamide.
Reaction Schemes A and steps and The above prepared amide (0.2 g, 0.4 mmol) is deprotected in a manner analogous to that described previously in example step B by treatment with HCO 2 H. The free amine is then coupled with N-[(3-Pyridinylmethoxy)carbonyl]-L-valine (0.38 g, 1.5 mmol), prepared in example 8, under standard coupling conditions in a manner analogous to that described previously to provide after recrystallization from ethyl acetate, 0.09 g of title compound as a white powder: mp 201-210 0 C; 1 H NMR (DMSO-d 6 6 9.48 (bs, 1 8.79 1 8.65 2 8.02-7.85 3 7.39-7.65 9 H), 7.24 2 H, J 8.5 Hz), 6.96 2 H, J 8.5 Hz), 6.48 1 H, J 10.4 Hz), 5.25 2 H, J 3.8 Hz), 5.14 (s, 2 4.63-4.55 2 4.36 1 4.17 1 H), 3.91 (dd, 1 H, J 7.8, 6.0 Hz) 3.14-3.03 1 2.81- 2.55 1 1.95 1 0.81 (dd, 6 H, J 9.1, Hz); 19 F NMR (DMSO-d 6 6 -110.17 (dd, J 254, 11 Hz), 119.79 (dd, J 247, 16 Hz).
Step C Preparation of final title compound.
Reaction Scheme step The above prepared alcohol (0.08 g, 0.1 mmol) is oxidized under Swern conditions in an analogous manner to that described previously, such as example 1, step G, from 2 M oxalyl chloride/CH 2 Cl 2 (0.6 mL, 1 mmol), DMSO (0.19 mL, 2.6 mmol), and diisopropylethylamine (0.41 mL, 2.4 mmol), 0.04 g to provide the final title compound as a tan powder: mp WO 96/02499 WO 9602499PCTIUS95/0724 1 0 C; IR (KBr) vmax 3401, 3306, 1699, 1599, 1437, 1298, 1242, 1178, 1113, 1026 cm- 1 'H NMR (CDCl 3 6 8.63-8.52 (in, 2 8.42 and 8.19, and 8.05 (3m, 2 H total), 7.77-7.62 (in, 2 7.46-7.22 (in, 9 7.18 1 H, J 5.5 Hz), 7.05 2 H, J 8.2 Hz), 6.89 2 H, J 8.2 Hz), 5.35 and 5.22 (2d, 1 H total, J 8.5 Hz), 5.10 2 5.05 2 4.87-4.66 (in, 1 4.60-4.48 (in, 1 3.81 (dd, 1 H, J 9.2, 6.7 Hz), 3.32 and 3.27 (2d, 1 H total, J 5.2 and 4.8 Hz), 3.18 and 3.13 (2d, 1 H total, J 9.2 and 8.4 Hz) 1.91 (mn, 1 1.31 1 H, J 6.8 Hz), 0.73 and 0.78 and 0.82 (3d, 5 H total, J 6.8 Hz); 19 F NMR (CDC1 3 6 -110.48 J 276 Hz), -113.38 -113.51 J =276 Hz), Hydrate: -118.75 J 254 Hz), -120.75 J =255 Hz); mass spectrum, m/z 674 460, 348, 280, 272, 252, 207, 138, 110 (100), 92.
Example 14 Preparation of tl-[ F[3,3-Difluoro-2,4-dioxo-l-[[4- (phenylmethoxy )phenyl ]-methyl] pyridinylmethyl)aminolbutyllamino]carbonyl]-2methylpropyl]-carbamic Acid, 3-Pyridinylmethyl Ester.
OBn
N
25 N
CF
2 NH r- 0 0 0 Step A Preparation of (1 3 ]-2,4,5-Trideoxv-4-[[(1,1dimethylethoxy) carbonyl 1-amino] 2-difluoro-5-[4- (phenylmethoxy)-phenyl 3-pyridinylmethyl)-L-glyceropentanamide.
Reaction Scheme A, step The title compound is prepared in a manner analogous to that described in example 13, step A from 4-tert-butoxycarbonylamino-2,2-difluoro-3- WO 96/02499 PCT/US95/07241 -76hydroxy-5-(4-benzyloxy)phenylpentanoic acid, ethyl ester g, 3.1 mmol) prepared in example 1 step B, and 3-(aminomethyl)pyridine (0.38 mL, 3.8 mmol). The precipitated solids are washed with ether and are recrystallized from ethyl acetate to provide 0.80g of title compound as a white powder: mp 158-160 0 C; 1 H NMR (CDC1 3 6 8.55 (bs, 2 7.7 1 H, J 7.2 Hz), 7.5-7.2 8 7.1 2 H, J 8.4 Hz), 6.9 2 H, J 8.3 Hz), 5.0 2 4.9 1 H, J 9.8 Hz), 4.5 2 H, J 2.6 Hz), 4.1-3.9 2 3.0-2.8 2 1.4 9 19 F NMR (DMSO-d 6 6 -110.84 J 272 Hz), -122.56 (dd, J 273, 21 Hz); mass spectrum, m/z 542 (M 486, 442, 424, 344, 244, 224, 197, 109, 91.
Step B Preparation of [3C,4(S)]-2,4,5-Trideoxy-2,2-difluoro-4-[[3methyl-l-oxo-2-[[(3pyridinylmethoxy)carbonyl]amino]butyl]amino]-5-[4-(phenylmethoxy)phenyl]-N-(3-pyridinylmethyl)-L-glyceropentanamide.
Reaction Schemes A and steps and The above prepared amide (0.27 g, 0.49 mmol) is treated with 97%
HCO
2 H. The reaction mixture became homogeneous within minutes. The solution is allowed to stir at room temperature for 3 hours. It is then concentrated under vacuum. The yellow oil is then dissolved in ethyl acetate, washed twice with aqueous 1 N sodium bicarbonate, dried over anhydrous magnesium sulfate, filtered and concentrated under vacuum to provide the corresponding free amine as a yellow solid (0.18 g, 0.41 mmol). This is dissolved in a 1:1 mixture of CH 2 C1 2 DMF (6 mL). To this stirring solution at room temperature is added HOBT (0.07 g, mmol), NMM (0.05 mL, 0.5 mmol), pyridinylmethoxy)carbonyl]-L-valine (0.13 g, 0.52 mmol), prepared in example 8 and EDC (0.1 g, 0.5 mmol) in that order. After 16 hours, the mixture is diluted with CH 2 C12 and washed with water. A white precipitate separates, WO 96/02499 PCT/US95/07241 -77which is filtered, washed several times with ether, and recrystallized from ethyl acetate to provide 0.12 g of title compound as a white powder: mp 195-196 0 C; 1 H NMR (DMSO-d 6 6 8.7-8.4 4 7.75 1 H, J 10.4 Hz), 7.65 1 H, J 11.2 Hz), 7.5-7.3 (m 7 7.15 2 H, J 8.4 Hz), 6.9 2 H, J 8.4 Hz), 5.12 2 5.08 2 4.5-4.2 3 4.1-3.9 2 2.8-2.6 2 2.0 1 0.8 6 19 F NMR (DMSO-d 6 6 -111.2 J 257 Hz), -122.05 (dd, J 250, 20 Hz); mass spectrum, m/z 676 (M 567, 424, 150, 138, 110 (100), 92. Anal. Calcd for C 36
H
39
F
2
N
5 0 6 C, 63.31; H, 5.87; N, 10.25. Found: C, 63.07; H, 5.90; N, 10.23.
Step C Preparation of final title compound.
Reaction Scheme step The above prepared alcohol is oxidized under Swern conditions in a manner analogous to example 1 as follows: To a slightly cloudy solution of the above prepared alcohol (0.08 g, 0.12 mmol) in CH 2 C1 2 (2 mL) and DMSO (0.21 mL) at -45 0 C is added 2M oxalyl chloride/CH 2 Cl 2 (0.6 mL, 1 mmol) over 5 minutes. The solution is allowed to stir at -35 to -45 0 C for 2.5 hours and is then cooled to -78 0 C. Diisopropylethylamine (0.42 mL, 2.4 mmol) is added over 5 minutes. The solution is allowed to warm to room temperature over 1.5 hours, is diluted with CH 2 C12 and washed with water. The yellow precipitate which forms is filtered and washed three times with ether to provide 0.04 g of final title compound as a tan powder: mp 103-105C; IR (KBr) Vmax 3422, 3295, 2965, 1697, 1649, 1537, 1512, 1244 cm- 1 1 H NMR (DMSO-d 6 6 9.19 (bs, 1 8.61-8.43 4 7.75-7.59 2 H), 7.45-7.28 7 7.09 2 H, J 8.5 Hz), 6.81 2 H, J 8.5 Hz), 5.09-4.98 4 4.50-4.26 3 H), 3.90-3.61 2 3.12 2 2.55 1 1.79 (m, 1 1.27 5 H, J 8.6 Hz), 0.77 1 H, J 6.2 Hz); 19 F NMR (DMSO-d 6 6 minor diastereomer: -111.34 (apparent doublet, J 184 Hz), major diastereomer: -113.24 J WO 96/02499 PCT/US95/07241 -78- 254 Hz), -117.56 J 254 Hz); mass spectrum, m/z 674 (M 565, 460, 252, 207, 143, 138, 110 (100), 92.
Anal. Calcd for C 36
H
37
F
2
N
5 0 6 .2H 2 0: C, 60.93; H, 5.78; N, 9.87. Found: C, 60.00; H, 6.15; N, 9.56.
Example Preparation of [1-[[[3,3-Difluoro-2,4-dioxo-l-[[4- (phenylmethoxy)-phenyl]-methyl]-4-[(2pyridinylmethyl)amino]butyl]-amino]carbonyl]- 2methylpropyl]-carbamic Acid, 2-Pyridinylmethyl Ester.
OBn o o 0 0 0 Step A 2 Preparation of N-[(2-Pyridinylmethoxy)carbonyl]-L-valine Methyl Ester.
In a manner analogous to the preparation of pyridinylmethoxy)carbonyl]-L-valine in example 8, the title compound is prepared from L-valine methyl ester 2 hydrochloride (5.0 g, 30 mmol), triphosgene (4.66 g, 15.7 mmol), and (n-Bu) 3 N (0.05 mL, 0.4 mmol); however, 2-pyridylcarbinol (10.0 mL, 100 mmol, 3.3 equivalents) is added dropwise to the intermediate isocyanate. Flash chromatography as in example 8 provides 2.63 g of title compound as a yellow oil: IR (neat) vmax 3345, 2965, 1726, 1533, 1439, 1314, 1271, 1236, 1213, 1105 cm- 1 1 H NMR (CDCl 3 6 8.61 1 H, J 4.6 Hz), 7.7 (td, 1 H, J 7.6, 1.8 Hz), 7.38 1 H, J 7.7 Hz), 7.27 (dd, 1 H, J 7.3, 4.6 Hz), 5.45 1 H, J 8.9 Hz), 5.25 2 4.35 (dd, 1 H, J 9.2, 4.7 Hz), 3.75 3 2.24 1 H), 3 H, J 6.8 Hz), 0.9 d, 3 H, J 6.8 Hz); 13 C NMR (CDC1 3 6 172.4, 156.2, 155.9, 149.4, 136.7, 122.7, 121.6, WO 96/02499 WO 9602499PCTJUS95/0724 1 -79- 67.4, 59.1, 52.1, 31.2, 18.9, 17.5; mass spectrum, iu/z 267 1, 100), 136, 110.
Anal. Calcd for C 1 3
H
18
N
2 0 4 C, 58.65; H, 6.77; N, 10.53.
Found: C, 58.25; H, 6.86; N, 10.46.
Step B Preparation of (2-Pyridinylmethoxy)carbonyl]-L-valine.
Hydrolysis of the above ester (2.5 g, 9.4 mmol) with LiOH*H 2 0 (0.79 g, 18.8 mmol) in CH 3 OH in a manner analogous to that described previously in example 8 provides 2.0 g of the title compound as a white solid: IR (KBr) Vmax 3366, 3065, 2963, 2936, 1722, 1616, 1602, 1577, 1526, 1433, 1279, 1250, 1219, 1109, 764, 627 cnC 1 1 H NMR (DMSO-d 6 6 8.58 1 H, J 4.7 Hz), 7.96 1 H, J 8.0 Hz), 7.6 1 H, J =8.0 Hz), 7.43 1 H, J =7.5 Hz), 7.4 (dd, 1 H, J 5.2 Hz), 5.15 2 3.9 (dd, 1 H, J 5.9 Hz), 2.1 (in, 1 0.92 (dd, 6 H, J 6.6, 3.7 Hz); 13 c NMR (DMS0-l 6 6 173.1, 156.3, 156.2, 148.6, 137.4, 123.0, 121.4, 65.9, 59.6, 29.5, 19.1, 18.0; mass spectrum, m/z 253 1) 237, 209, 136 (100), 109, 92, Step C Preparation of 4(S)]-2,4,5-Trideoxy-2,2-difluoro-4t(3-methyl-l-oxo-2-( (2pyridinvlmethoxy)carbonyllaminolbutyl~amino]-5-[4-(phenylmethoxy )phenyl 1-N- (2-pyridinylmethyl )-L-glyceropentanamide.
Reaction Schemes A and steps and The [1 3&]-2,4,5-trideoxy-4-[ f(1,1dimethylethoxy)carbonyl]-amino]-2,2-difluoro-5-[4- (phenylmethoxy)-phenyll-N-( 2-pyridinylmethyl)-L-glyceropentonamide (0.34 g, 0.63 inmol) prepared in example 13, step A, is deprotected as in example 13, step and coupled with the above prepared acid (0.21 g, 0.82 inmol) under standard coupling conditions in a manner analogous to that described previously to provide 0.14g of title compound as a white powder: mp 224-225.5 0 C; IR Vmax WO 96/02499 PCT/US95/07241 3291, 1697, 1680, 1657, 1618, 1599, 1572, 1537, 1512, 1454, 1439, 1398, 1341, 1300, 1244, 700 cm-l; 1 H NMR (DMSO-d 6 6 9.31 (bs, 1 7.82-7.71 3H), 7.48-7.23 10 H), 7.10 2 H, J 8.6 Hz), 6.82 2 H, J 8.6 Hz), 5.05- 4.95 4 4.47 2 H, J 5.8 Hz), 4.23 1 H), 4.04 1 3.79 1 H, J 6.0 Hz), 4.40 and 3.69 and 3.14 (3m, 1 H total), 2.94 1 H, J 13.0 Hz), 2.63 (dd, 1 H, J 14.6, 10.8 Hz), 1.83 1 0.70 (dd, 5 H, J 9.2, 7.1 Hz), 0.57 and 0.47 (2d, 1 H total, J 6.9 Hz); 13 C NMR (DMSO-d 6 6 170.1, 157.4, 156.7, 156.6, 155.7, 148.97, 148.8, 137.2, 136.8, 130.76, 130.21, 128.4, 127.7, 127.5, 122.75, 122.25, 121.0, 120.8, 114.2, 69.0, 66.2, 60.38, 49.9, 44.1, 44.0, 34.0, 30.3, 19.2, 17.95; 19 F NMR (DMSO-d 6 6 major diastereomer: -110.07 J 248 Hz), 119.9 J 252 Hz), minor diastereomer: -110.03 (dm, J 250 Hz), -118.34 (dm, J 250 Hz), -111.72 (dm, J 250 Hz), -123.5 (dm, J 250 Hz); mass spectrum, m/z 676 (M 595, 567, 110, 92 (100).
Step D Preparation of final title compound.
Reaction Scheme step The above alcohol (0.13 g, 0.19 mmol) is oxidized under Swern condition in a manner analogous to that described previously in example 1 to provide after flash chromatography (8:1 CH 2 Cl 2
/C
2
H
5 0H) 0.04 g of a 1:1 mixture of the final title compound as a tan powder: mp 140-142 0 C; IR (KBr) vmax 3304, 1695, 1668, 1537, 1514, 1439, 1296, 1244, 1109, 1042 cm-l; 1 H NMR (DMSOd 6 6 9.09 and 8.85 and 8.09 m, d, 1 H total), 8.56 (m, 1 8.17 1 7.78-7.65 2 7.45-7.15 7.05 2 H, J 8.6 Hz), 6.89 2 H, J 8.6 Hz), 5.56 and 5.43 (2d, 1 H total, J 9.2, 9.2 Hz), 5.30-4.95 5 4.84-4.45 2 3.85-3.7 1 3.32-3.27 (2d,l H, J 4.9, 4.0 Hz), 3.16-2.80 (2m, 1 1.90-1.70 1 0.83-0.75 5 0.56 1 H, J 6.8 Hz); 1 9
F
NMR (DMSO-d 6 6 -110.07 J 275 Hz), -113.25 (apparent d, J 9 Hz), -113.45 J 276 Hz), Hydrate: -118.5 (d, WO 96/02499 PCT/US95/0724 I -81- J 257 Hz), -121.00 J 253 Hz); mass spectrum, m/z 674 460, 143, 110, 92.
WO 96/02499 PCTIUS95/07241 -82- Example 16 Preparation of [1-([[3,3-Difluoro-2,4-dioxo-l-[[4- (phenylmethoxy)-phenyl]-methyl]-4-[(3pyridinylmethyl)amino]butyl]-amino]carbonyll- 2methylpropyl]-carbamic Acid, 2-Pyridinylmethyl Ester.
OBn 0~ N 0 NH CF 2
NH
Step A Preparation of 4(S)]-2,4,5-Trideoxy-2,2-difluoro-4- [f3-methyl-l-oxo-2-[ 1(2pyridinylmethoxy)carbonyl amino]butyl amino1-5-[4-(phenylmethoxy)phenyl]-N-(3-pyridinylmethyl)-L-glyceropentanamide.
Reaction Schemes A and steps and The [1 3 I-2,4,5-trideoxy-4-[[(l,l-dimethylethoxy)carbonyllamino]-2,2-difluoro-5-[4-(phenylmethoxy)-phenyl)-N-(3pyridinylmethyl)-L-glycero-pentonamide (0.357 g, 0.709 mmol) prepared in example 14, step A, is deprotected as in example 14, step and coupled with N- [(2-pyridinylmethoxy)carbonyll-L-valine (0.157 g, 0.620 mmol) prepared in example 15, under standard coupling conditions described previously to provide after treating the crude material with hot CH 3 OH, filtering, and concentrating under vacuum, 180 mg of title compound as a white powder: IR (KBr) Vmax 3298, 1697, 1680, 1659, 1537, 1512, 1298, 1244, 1103 cm- 1 1 H N4R (DMSO-d 6 6 9.36- 9.32 1 8.52-8.46 3 7.82-7.68 3 H), 7.41-7.26 7 7.19 1 H, J 9.6 Hz), 7.07 2 H, J 8.4 Hz), 6.81 2 H, J 8.4 Hz), 6.27 1 H, J 7.2 Hz), 5.12 1 H, J 13.5 Hz), 5.06 1 H, J 13.5 Hz), 5.02 1 H, J 12.0 Hz), 4.97 1 H, J WO 96/02499 PCT/US95/07241 -83- 12.0 Hz), 4.44 (dd, 1 H, J 15.3, 6.0 Hz), 4.35 (dd, 1 H, J 15.3, 5.7 Hz), 4.24-4.15 1 4.11 (dd, 1 H, J 10.5, 5.1 Hz), 4.06-3.95 1 3.79 (dd, 1 H, J 9.3, 6.9 Hz), 2.93-2.88 1 2.60 (dd, 1 H, J 13.8, 11.1 Hz), 1.82-1.77 1 0.71 3H, J 6.9 Hz), 0.68 (d, 3H, J 6.9 Hz); 19 F NMR (DMSO-d 6 6 -109.87 (dd, J 252, 8 Hz), -120.07 (dd, J 253, 17 Hz); mass spectrum, m/z 704 (M 29), 676 (M 424, 241, 138, 110, 92 (100).
Anal. Calcd for C3 6
H
39
F
2
N
5 0 6 C, 63.99; H, 5.82; N, 10.36.
Found: C, 60.62; H, 5.97; N, 9.83.
Step B Preparation of final title compound.
Reaction Scheme step In a manner analogous to that described previously in example 1, the above prepared alcohol (152 mg, 0.225 mmol) is oxidized under Swern conditions to provide after flash chromatography (19:1 CHC1 3
/CH
3 0H) 104 mg of final title compound which is recrystallized twice from CH2Cl2/pentane to provide 34 mg of final title compound as a cream powder: mp 97- 102 0 C; IR (KBr) Vmax 3405, 3324, 1699, 1534, 1512, 1437, 1242, 1178, 1113 cm-l; 1 H NMR (CDC1 3 6 8.56-8.51 3H), 7.71-7.62 2 7.43-7.19 10 7.10-7.05 2 6.91-6.83 2 6.48-6.37 1 5.48-5.10 (m, 3H), 5.02 and 5.00 (2s, 2 H total), 4.61-4.46 2 H), 3.90 and 3.87-3.81 (dd and m, 1 H total, J 8.7, 6.1 Hz), 3.36-3.23 1 2.91-2.73 1 H total), 2.07-1.88 (m, 1 0.86 and 0.79 and 0.76 and 0.75 and 0.61 and 0.59 (6d, 6 H total, J 6.6 Hz); 19 F NMR (CDC1 3 6 -111.65
J
277 Hz), -111.73 J 276 Hz), -113.15 J 276 Hz), -113.20 J 277 Hz), Hydrate: -115.64 J 257 Hz), -116.55 J 256 Hz), -120.42 J 256 Hz), 121.55 J 256 Hz); mass spectrum, m/z 702 29), 674 (M 138, 110, 92 (100). Anal. Calcd for C3 6
H
37
F
2
N
5 0 6 C, 64.18; H, 5.54; N, 11.87. Found: C, 61.96; H, 5.80; N, 10.38.
WO 96/02499 PCT/US95/07241 -84- Example 17 Preparation of N-[4-(N-{2-pyridylmethyl}oxycarbonyl-Lvalyl)amino-2,2-difluoro-1,3-dioxo-5-(4-benzyloxy)phenylpentyl]-O-(3-pyridylmethyl)-D-valinol.
N OBn N
O
Step A Preparation of N-trityl-D-valinol.
A solution of D-valinol (4.95 g, 48.06 mmol), triethylamine (7.4 mL, 52.87 mmol) and trityl chloride (14.74 g, 52.87 mmol) in dry dichloromethane (75 mL) is stirred for 17 hours at room temperature. The organic solution is washed with water (2 x 75 mL), dried over anhydrous sodium sulfate, filtered and concentrated under vacuum. The resulting oil is purified by flash chromatography (silica gel, ethyl acetate/petroleum ether, 15/85) to provide the title compound (13.5 g, Rf=0.45 (ethyl acetate petroleum ether, 15/85).
Step B Preparation of N-trityl-O-3-pyridylmethyl-D-valinol.
Under a nitrogen atmosphere, to a suspension of sodium hydride (1.3 g, 30 mmol, 55% dispersion in oil, previously washed twice with pentane) in dry DMF (3 mL) is added with stirring, a solution of N-trityl-D-valinol (3.45 g, mmol) in DMF (23 mL). The reaction is stirred for minutes at room temperature and then cooled to 0°C and tetrabutylammonium iodide (0.37 g, 1 mmol) is added. Then 3-picolyl chloride HC1 (1.81 g, 11 mmol) is added to the reaction in portions over 5 minutes. After addition is WO 96/02499 PCT/US95/07241 complete, the cooling bath is removed and the mixture allowed to stirr for 17 hours at room temperature. The reaction mixture is then cooled with an ice bath and hydrolyzed with water (100 mL). The mixture is then extracted with ethyl acetate (2 x 100 mL). The organic extracts are washed with water (2 x 50 mL), combined, dried over anhydrous sodium sulfate, filtered and concentrated under vacuum. The resulting yellow oil is purified by flash chromatography (silica gel, dichloromethane/ethyl acetate, 9/1, Rf=0.42) to provide the title compound (3.4 g, 78%) as an oil.
Step C Preparation of O- 3 -pyridylmethyl-D-valinol.
A solution of N-trityl-O-3-pyridylmethyl-D-valinol (3.63 g, 8.3 mmol) in formic acid (30 mL) is kept for hours at room temperature. The formic acid is removed under vacuum and the residue is dissolved in water (100 mL). The mixture is extracted with ethyl acetate (100 mL, 50 mL) to remove the trityl alcohol. The aqueous phase is then made basic with saturated sodium carbonate (50 mL) and 4N sodium hydroxide (3 mL). The aqueous mixture is then extracted with ethyl acetate (4 x 50 mL). The organic extracts are rinsed with brine (2 x 50 mL), combined, dried over sodium sulfate, filtered and concentrated under vacuum to provide the title compound (1.32 g, 82%) Rf=0.12 (silica gel, dichloromethane/methanol, 8/2).
Step D Preparation of N-[ 4 -tert-butoxycarbonylamino-2,2-difluoro- 3-hydroxy-l-oxo-5-(4-benzyloxy)phenyl-pentyl]-0-(3pyridylmethyl)-D-valinol.
Reaction Scheme A, step A solution of the ester (1.14 g, 2.38 mmol, prepared in example 1, step B) and of
O-
3 -pyridylmethyl-D-valinol (1.32 g, 6.8 mmol, prepared in step C above) in dry tetrahydrofuran (1.5 mL) is heated for 2 days under reflux. After cooling, the reaction mixture WO 96/02499 PCT/US95/07241 -86is diluted with ethyl acetate (5 mL), pentane (10 mL) and the precipitate is collected by filtration. The filtrate is rinsed with pentane and recrystallized from dichloromethane/methanol/pentane to provide the title compound (0.8 g, 54%) as a white solid, Rf=0.5 (silica gel, ethyl acetate); mass spectrum, m/z 628 Step E Preparation of N-[4-amino-2,2-difluoro-3-hydroxy-l-oxo-5- (N-(benzyloxy)phenyl-pentyl]-O-(3-pyridylmethyl)-D-valinol.
Reaction Scheme A, step The title compound is obtained in 91% yield following the deprotection procedure in an analogous manner described in Example 17, step C; mass spectrum, m/z 528 Step F Preparation of N-[4-(N-{2-pyridylmethyl)oxycarbonyl-Lvalyl)amino-2,2-difluoro-3-hydroxy-l-oxo-5-(4benzyloxy)phenyl-pentyl]-0-(3-pyridylmethyl)-D-valinol.
Reaction Scheme step To a solution of N-(2pyridylmethyloxycarbonyl)-L-valine (0.101 g, 0.4 mmol, prepared in Example 15, step in anhydrous DMF (2 mL) are added under an atmosphere of nitrogen, Nhydroxybenztriazole hydrate (0.115 g, 0.4 mmol), 1-ethyl- 3(3-dimethylaminopropyl)carbodiimide (0.085 g, 0.44 mmol) and DMF (1 mL). The reaction mixture is allowed to stir for 30 minutes at room temperature and N-[4-amino-2,2difluoro- 3 (3-pyridylmethyl)-D-valinol (0.211 g, 0.4 mmol, prepared above) in DMF (1 mL) is added. The reaction is allowed to stir for 15 hours, then diluted with ethyl acetate (80 mL) and washed with water (2 x 80 mL). The aqueous rinses are extracted with ethyl acetate (80 mL). The combined organic extracts are dried over anhydrous sodium sulfate, filtered and concentrated under vacuum. The residue is purified by flash chromatography (2x, silica gel, WO 96/02499 PCTIUS95/07241 -87dichloromethane/ethanol, 95/5, Rf=0.15) to provide the title compound (0.170 g, mass spectrum, m/z 762 Step G Preparation of final title compound.
Reaction Scheme step To a solution of oxalyl chloride (0.195 mL, 2.23 mmol) in anhydrous dichloromethane mL) at -60 0 C is added slowly under an atmosphere of nitrogen, freshly distilled dimethyl sulfoxide (0.316 mL, 4.46 mmol) in dichloromethane (1.5 mL). After 10 minutes of stirring at -60 0 C, the temperature is allowed to rise to 0 C. A mixture of a solution of the alcohol prepared above (0.170 g, 0.223 mmol) in dichloromethane (7 mL) and dimethylsulfoxide (0.5 mL) is added to the reaction dropwise. The reaction is then stirred for 1.75 hours at 0 C and then cooled to -78 0 C. Diisopropylethylamine (0.93 mL, 6.69 mmol) is added and the reaction is stirred an additional 10 minutes. The reaction is then allowed to warm to room temperature and is diluted with dichloromethane (25 mL). The reaction mixture is washed with water (2 x 25 mL). The aqueous washes are extracted with dichloromethane (25 mL). The combined organic extracts are dried over anhydrous sodium sulfate, filtered and concentrated under vacuum. The residue is purified by flash chromatography (silica gel, ethyl acetate, Rf=0.17) followed by recrystallization from dichloromethane/pentane to provide the title compound (0.120 g, mass spectrum, m/z 760 1 H NMR (DMSO-d 6 6 9.12-8.96 (m, 1H), 8.66-8.55 2H), 7.96-7.85 1H), 7.85-7.75 (m, 1H), 7.60-7.35 7H), 7.30-7.19 2H), 7.08-6.83 (m, 2H), 5.20 2H), 5.17-5.05 3H), 4.65-4.53 2H), 4.07-3.87 (2m, 2H), 3.72-3.60 2H), 3.28-3.14 and 2.86- 2.78 2H), 2.09-1.83 2H), 1.27-0.67 12H); 19 F NMR (DMSO-d 6 T=60 0 C) 6 (C 6
F
e 52.94 J=269 Hz), 52.30 (d, J=270 Hz), 51.25 J=270 Hz), 50.35 J=269 Hz).
Anal. Calcd for C 41
H
47
N
5 0 7
F
2 0.25 H 2 0: C, 64.43; H, 6.26; N, 9.16; WO 96/02499 PCT/US95/07241 -88- Found C, 64.24; H, 6.25; N, 9.01.
Example 18 Preparation N-[4-(N-{3-pyridylmethyl)oxycarbonyl-Lvalyl)amino-2,2-difluoro-1,3-dioxo-5-(4-benzyloxy)phenylpentyl]-O-(3-pyridylmethyl)-D-valinol.
OBn
N
O
Y
Step
A
Preparation of N-(4-(N-(3-pyridylmethyl}oxycarbonyl-Lvalyl)amino-2,2-difluoro-3-hydroxy-l-oxo-5-(4benzyloxy)phenyl-pentyl]-0-(3-pyridylmethyl)-D-valinol.
Reaction Scheme step The title compound is 2 prepared in a manner analogous to the coupling procedure described in Example 17, Step F, utilizing the amine prepared in Example 17, Step E, and the acid prepared in Example 8, Step A. The title compound is obtained in 84% yield, Rf=0.43 (silica gel, dichloromethane/ethanol, 95/5); mass spectrum, m/z 762 Anal. Calcd for C 41
H
49
N
5 0 7
F
2 *0.5 H 2 0: C, 63.88; H, 6.54; N, 9.08; Found C, 63.87; H, 6.47; N, 9.06.
3 Step B Preparation of Final Title Compound.
Reaction Scheme step The title compound is prepared in a manner analogous to the Swern Oxidation described in Example 17, Reaction Scheme step 3 utilizing the above alcohol. The title compound is obtained in 50% yield, Rf=0.1 (silica gel, ethyl acetate); 1 H NMR (DMSO-d 6 6 9.04-8.90 1H), 8.71-8.50 3H), 7.90-7.70 2H), 7.60-6.85 12H), 5.27-5.02 WO 96/02499 PCTUS95/07241 -89- 4.50-4.66 (mn, 2H), 3.85-4.07 (mn, 2H), 3.72-3.57 (mn, 2H), 3.30-3.12 and 2.85-2.65 (mn, 2H), 2.05-1.8 (mn, 2H), 1.20- 0.59 (mn, 12H); 1 9 F NMR (DMSO-d 6 6 (C 6
F
6 52.93 J=268 Hz), 52.15 J=269 Hz), 50.92 J=269 Hz), 49.68 (d, J=268 Hz); hydrates, 49.31 J=254 Hz), 48.31 J=254 Hz), 46.22 J=254 Hz), 44.68 J=254 Hz).
Anal. Calcd for C 4 1
H
47
N
5 0 7
F
2
H
2 0: C, 63.31; H, 6.35; N, 9.00; Found C, 62.56; H, 6.26; N, 8.66.
Example 19 Preparation of N-f 4-(N-{3-pyridylnethyl~oxycarbonylLvalyl)amino-2,2-difluoro-,3dioxo5.(4-.benzvloxv)phenyl- Dentyl 1-0-methyl -D-valinol.
N.OBn 0
N
0 N H CF 2
NH
YI/ K H<~b
H
0 0 0 Step A Preparation of N-tert-butoxycarbonyl-D-valinol.
A solution of D-valinol (5.1 g, 49.4 mmol) and di-tertbutyldicarbonate (10.9 g, 50 nunol), in methanol (60 mL) is stirred for 17 hours at room temperature. The reaction is concentrated under vacuum and the residue is purified by flash chromatography (silica gel, ethyl acetate/petroleum ether, 3/7, Rf=0.37) to provide the title compound in quantitative yield (10.07 g) as a colorless oil; mass spectrum, m/z 204 WO 96/02499 PCT/US95/07241 Step B Preparation of N-tert-Butoxycarbonyl-O-methyl-D-valinol.
To a solution of N-tert-butoxycarbonyl-D-valinol (5.075 g, 25 mmol) and methyl iodide (27.5 mL of a 1M solution in tetrahydrofuran, 27.5 mmol) in dry tetrahydrofuran (45 mL) under an atmosphere of nitrogen at 0°C, is added with stirring, potassium-tert-butoxide (3.086 g, 27.5 mmol) in 3 portions. The reaction mixture is kept at room temperature for 5.5 hours, then hydrolyzed with a saturated solution of sodium chloride and extracted with ethyl acetate (2 x 300 mL). The organic extracts are washed with brine (2 x 300 mL), combined, dried over anhydrous sodium sulfate, filtered and concentrated under vacuum. The residue is purified by flash chromatography (silica gel, ethyl acetate/petroleum ether, 1/9, Rf=0.25) to provide the title compound (2.93 g, 54%) as an oil.
Step C Preparation of O-methyl-D-valinol.
A solution of N-tert-butoxycarbonyl-0-methyl-D-valinol (2.93 g, 13.5 mmol) in dry ether saturated with hydrogen chloride (11 mL) is stirred at room temperature for hours. The reaction is then concentrated under vacuum.
Pentane (50 mL) is added to the residue, followed by addition of diethylamine (7 mL). The mixture is then stirred for 10 minutes and the solid is removed by filtration. The filtrate is concentrated under vacuum(150 bars) at room temperature to provide the title compound (1.30 g, 82%) as a colorless oil.
Step D Preparation of N-[ 4 -tert-butoxycarbonylamino-2,2-difluoro- 3-hydroxy-l-oxo-5-(4-benzyloxy)phenyl-pentyl]-O-methyl-Dvalinol.
Reaction Scheme A, step The title compound is prepared in a manner analogous to the procedure of Example 17, Step D, from the ester of Example 1, step B and the WO 96/02499 WO 9602499PCTIUS95/0724 1 -91above prepared O-methyl-D-valinol followed by flash chromatography (78% yield); Rf=0.19 (minor) and 0.09 (major) (silica gel, ethyl acetate/petroleum ether, 35/65); mass spectrum, m/z 551 568 (MNH 4 Step E Preparation of N- [4-amino-2 ,2-difluoro-3-hydroxy-l-oxo-5- (4-benzyloxy)phenyl-pentyl ]-0-methyl-D-valinol.
Reaction Scheme A, step The title compound is prepared in a manner analogous to the deprotection procedure of Example 17, step C, from N-114-tertbutoxycarbonylamino-2 ,2-difluoro-3-hydroxy-l-oxo-5- (4butyloxy )phenyl-pentyl ]-O-methyl-D-valinol prepared above (quantitative yield); mass spectrum, m/z 451
(MH
4 Step F Preparation of N-[4-(N--{3-pyridylmethyl~oxycarbonyl-Lvalyl )amino-2, 2-difluoro-3-hydroxy-l-oxo-5-(C4benzyloxy)phenyl-pentyl ]-O-methyl-D-valinol.
Reaction Scheme step The title compound is prepared in a manner analogous to the coupling method described in Example 17, step F, from N-i14-amino-2,2- 4-benzyloxy)phenyl-pentyl methyl-D-valinol prepared above and the acid prepared in Example 8, Step A. Purification by crystallization (ethyl acetate plus 10% ethanol/pentane) provides the title compound in 58% yield; mass spectrum, m/z 685 (MH11).
Step G Preparation of final title compound.
Reaction Scheme step The final title compound is prepared in a manner analogous to the Swern Oxidation described in Example 17, step G from pyridylmethyl~oxycarbonyl-L-valyl)amino-2,2-difluoro-3- (4-benzyloxy )phenyl-pentyl I-Q-methyl-Dvalinol prepared above (15% yield, Rf=0.l, silica gel, WO 96/02499 PCT/US95/07241 -92dichloromethane/ethyl acetate, 1 H NMR (DMSO-d 6 6 9.07-8.93 1H), 8.87-8.50 2H), 8.40-8.31 1H), 7.94-7.81 1H), 7.60-6.59 (several m, 11H), 5.29-5.01 5H), 4.10-3.78 2H), 3.58-3.43 2H), 3.35-3.25 (several s, 3H), 3.30-3.08 and 2.85-2.63 2H), 2.11- 1.72 2H), 1.04-0.48 12H); 19 F NMR (DMSO-d 6 6 (C 6 Fe) 53.27 J=267 Hz), 52.60 J=269 Hz), 51.32 J=269 Hz), 49.76 J=267 Hz), 49.89 J=254 Hz), 48.61 (d, J=254 Hz), 46.49 J=254 Hz), 44.73 (J=254 Hz); mass spectrum, m/z 683 Anal. Calcd for C 36
H
44
N
4 0 7
F
2 H20: C, 61.70; H, 6.62; N, 7.99; Found C, 62.07; H, 6.46; N, 7.84.
Example Preparation of N-{4-(N-{3-pyridylmethyl}oxycarbonyl-Lvalyl)amino-2,2-difluoro-l,3-dioxo-5-(4-benzyloxy)phenylpentyl]-O-(2-pyridylmethyl)-D-valinol.
OBn OBn 0N NH CF 2
NH
00 N Step A Preparation of N-trityl-O-2-pyridylmethyl-D-valinol.
The title compound is prepared in a manner analogous to the procedure described in Example 17, step B from the alcohol of Example 17, step A and 2-picolyl chloride HC1, in 81% yield, Rf=0.52 (silica gel, dichloromethane/ethyl acetate, 9/1).
Step
B
Preparation of O-2-pyridylmethyl-D-valinol.
The title compound is prepared in a manner analogous to the procedure described in Example 17, step C from N- WO 96/02499 WO 9602499PCTIUS95/07241 -93trityl-0-2--pyridylmethyl-D-valinol prepared above, in yield.
Step C Preparation of N-[ 4 -tert-butoxycarbonylamino-2,2.difluoro- 3-hydroxy--l-oxo-5-(C4-benzvloxy )phenyl-pentyl 2pyridylmethyl )-D-valinol.
Reaction Scheme A, step The title compound is prepared in a manner analogous to the procedure described in Example 17, Step D, from the ester of Example 1, step B and 0-2-pyridylmethyl-D-valinol, prepared above, in 63% yield, Rf=0.65 (silica gel, ethyl acetate).
Step D Preparation of N-[ 4 -amino-2,2-difluoro3royl..oxlox-5.
4 -benzyloxy)phenyl-pentvl
I-C-C
2 -pyridylmethyl)-D-valinol.
Reaction Scheme A, step The title compound is prepared in a manner analogous to the deprotection procedure described in Example 17, step C from N-[4-tertbutoxycarbonylamino-2, 2 -difluoro-3-hydroxy-l-oxo-5 (4benzyloxy )phenyl-pentyl 1-0- (2-pyr idylmethyl )-D-val inol, prepared above, in 91% yield which is used directly in the next step; mass spectrum, m/z 628 Step E Preparation of N-[4-(N-(37_pyridylmethylloxycarbonyl-L valyl )amino-2 ,2-difluoro-3-hydroxyloxo-... C4benzyloxy )phenyl-p~entyl 1-0-C2-pyridylmethyl )-D-valinol.
Reaction Scheme step The title compound is prepared in a manner analogous to the procedure described in Example 17, step F from the amine prepared above and the acid prepared in Example 8, Step A, in 53% yield, Rf=O-l4 (major) and 0.08 (minor) (silica gel, dichloromethane/ethanol, 95/5); mass spectrum, m/z 762
(MHII).
WO 96/02499 PCTIUS95/07241 -94- Step F Preparation of final title compound.
Reaction Scheme step The final title compound is prepared in a manner analogous to the Swern Oxidation procedure described in Example 17, step G from the above prepared alcohol, in 72% yield, Rf=0.09 (silica gel, ethyl acetate); 1 H NMR (CDC13) 6 8.65-8.53 1H), 7.79-7.64 (m, 2H), 7.53-6.80 (several m, 14H), 6.60-6.10 (several m, 2H), 5.57-5.33 1H), 5.30-5.10 3H), 4.99 2H), 4.68- 4.56 2H), 4.03-3.85 2H), 3.85-3.34 2H), 3.42- 3.24 and 2.97-2.75 2H), 2.13-1.88 2H), 1.05-0.57 12H); 19 F NMR (CDC13) 6 (CeFe) 50.26 J=271 Hz), 49.97 J=263 Hz), 49.18 J=261 Hz), 48.60 J=271 Hz), 45.17 J=252 Hz), 45.10 J=251 Hz), 41.77 (d, J=252 Hz), 40.91 J=251 Hz); mass spectrum, m/z 760 Anal. Calcd for C 41
H
47
N
5 0 7
F
2 C, 64.81; H, 6.23; N, 9.22; Found C, 62.91; H, 6.16; N, 8.66.
WO 96/02499 PCT/US95/07241 Example 21 Preparation of N-[4-(N-{2-pyridylmethyl~oxycarbolvl-Lvalyl)amino-2,2-difluoro-l,3-dioxo-5-(4-(3pyridylmethyl }oxy) phenyl-pentyl 1-0- (3-pyr idylmethyl valinol.
C
100 N ~0 NH CF 2
NXH
0 0 0 Step A Preparation of 4-tert-butoxycarbonylamino-2,2-difluoro- 3 hydroxy-5-(4-hydroxy)phenyl pentanoic acid, ethyl ester.
Reaction Scheme A, step A solution of 4-tertbutoxycarbonylamino-2, 2-difluoro-3-hydroxy-5- benzyloxy)phenyllpentanoic acid, ethyl ester (0.719 g, mmol) in ethanol (50 mL) is kept for 7.5 hours under an atmosphere of hydrogen in the presence of 10% palladium on charcoal (0.074 The hydrogen atmosphere is then exchanged with a nitrogen atmosphere, the suspension is filtered and the filtrate is concentrated under vacuum to provide the title compound (0.500g, Rf=0.51 (silica gel, petroleum ether/ethyl acetate, 1/1).
Step B Preparation of N-f 4-tert-butoxycarbonylamino-2 ,2-difluoro- 4-hydroxy)phenyl-pentyl]1-0- (3pyridylmethyl )-D-valinol.
Reaction Scheme A, step The title compound is prepared in a manner analogous to the procedure described in Example 19, step D, from the above prepared ester and WO 96/02499 PCT/US95/07241 -96the amine prepared in Example 17, step C, in 82% yield, Rf=0.46 (silica gel, ethyl acetate).
Step C Preparation of N-[4-tert-butoxycarbonylamino-2,2-difluoro- 3-hydroxy-l-oxo-5-(4-hydroxy)phenyl-pentyl]-O-(3pyridylmethyl)-D-valinol.
Reaction Scheme A, step A mixture of N-[4-tertbutoxycarbonylamino-2,2-difluoro-3-hydroxy-l-oxo-5-(4hydroxy)phenyl-pentyl]-O-(3-pyridylmethyl)-D-valinol (0.376 g, 0.7 mmol, prepared above), 3-picolyl chloride HC1 (0.161 g, 0.98 mmol), cesium carbonate (0.775 g, 2.38 mmol) and potassium iodide (0.016 g, 0.098 mmol) in anhydrous DMF (7 mL) is stirred for 66 hours under an atmosphere of nitrogen. The reaction mixture is diluted with ethyl acetate (50 mL) and washed with water (2 x 50 mL). The aqueous rinses are extracted with ethyl acetate (50 mL).
The combined organic extracts are dried over anhydrous sodium sulfate, filtered and concentrated under vacuum.
The residue is purified by flash chromatography (silica gel, ethyl acetate/methanol, 95/5, Rf=0.25) to provide the title compound (0.245 g, mass spectrum m/z 629 (MH Step D Preparation of N-[4-amino-2,2-difluoro-3-hydroxy-l-oxo-5- (4-(3-pyridylmethyl}oxy)phenyl-pentyl]-O-(3-pyridylmethyl)- D-valinol.
Reaction Scheme A, step The title compound is prepared in a manner analogous to the deprotection procedure described in Example 17, step C from N-[4-tertbutoxycarbonylamino-2, 2 -difluoro-3-hydroxy-l-oxo-5-(4hydroxy)phenyl-pentyl]-0-(3-pyridylmethyl)-D-valinol prepared above, in 84% yield; mass spectrum m/z 529 WO 96/02499 PCT/US95/07241 -97- Step E Preparation of N-[4-(2-pyridylmethyl)oxycarbonyl-Lvalyl)amino-2,2-difluoro-3-hydroxy-l-oxo-5-(4-{3pyridylmethyl}oxy)phenvl-pentyl]-O-(3-pyridylmethyl)-Dvalinol.
Reaction Scheme step The title compound is prepared in a manner analogous to the coupling procedure described in Example 17, step F, from the above prepared amine and the acid prepared in Example 15, step B, in 56% yield, Rf=0.20 (silica gel, dichloromethane/ethanol, 95/5); mass spectrum m/z 763 Step F Preparation of final title compound.
Reaction Scheme step The final title compound is prepared in a manner analogous to the Swern Oxidation procedure described in Example 17, step G from the above prepared alcohol, in 70% yield, Rf=0.19 (silica gel, ethyl acetate/acetone, mass spectrum m/z 761 1 H NMR (DMSO-d 6 6 9.11-8.97 1H), 8.80-8.50 5H), 8.50-8.30 1H), 8.13-7.70 4H), 7.69-6.60 8H), 5.40-5.00 5H), 4.71-4.49 2H), 4.10-3.80 2H), 3.80-3.51 2H), 3.51-3.30 and 2.86-2.60 2H), 2.12-1.72 (m, 2H), 1.17-0.47 12H); 19 F NMR (DMSO-d 6 6 (C 6
F
6 52.91 J=268 Hz), 52.11 J=269 Hz), 50.86 J=269 Hz), 50.42 J=268 Hz), 49.34 J=254 Hz), 48.39 J=254 Hz), 46.17 J=254 Hz), 44.63 J=254 Hz).
Anal. Calcd for C 40
H
46
N
6 0 7
F
2 C, 63.15; H, 6.09; N, 11.05; Found C, 62.26; H, 6.24; N, 10.76.
WO 96102499 PCTIUS95/07241 -98- Example 22 Preparation of N-[4-(N-{2-pyridylmethylloxycarbonyl-Lvalyl)amino-2,2-difluoro-1,3-dioxo-5-(4-benzyloxy)phenylpentyl 1-0-C2-pyridylmethyl )-D-valinol.
N. OBn 100 0NHCFY<H 0 0 CIN Step
A
Preparation of 4-f 2-pvridylmethyl}oxycarbonyl-Lvalyl )amino-2, 2-difluoro-3-hydroxy-l-oxo-5- (4benzyloxy)phenyl-pentyl]-O-( 2-pyridylmethyl)-D-valinol.
Reaction Scheme step The title compound is prepared in a manner analogous to the coupling procedure described in Example 17, step F from the amine prepared in Example 20, Reaction Scheme A, step and the acid prepared in Example 15, step B, in 57% yield, RfO0.17 (major) and 0.12 (minor) (silica gel, dichloromethane/ethanol, 95/5); mass spectrum m/z 762 Step B Preparation of final title compound.
Reaction Scheme step The final title compound prepared in a manner analogous to the Swern Oxidation procedure described in Example 17, step G from the above prepared alcohol, in 80% yield, Rf=0.22 (silica gel, ethyl acetate); mass spectrum m/z 760 1 H NMR (CDCl 3 6 8.63-8.48 (in, 2H), 7.77-7.60 (in, 211), 7.55-6.80 (several in, 351311), 6.49-6.02 (several mn, 2H), 5.55-5.30 (mn, 1H1), 5.30- 4.95 (several in, 5H1), 4.72-4.52 (mn, 2H), 4.05-3.82 (mn, 2H), 3.82-3.35 (in, 2H), 3.42-3.23 and 2.97-2.77 (in, 211), 2.13- 1.83 (in, 211), 1.05-0.54 (mn, 1211); 19 F NMR (CDC1 3 6 (COO 6 50.18 J=271 Hz), 49.87 J=277 Hz), 49.02 J=271 WO 96/02499 PCTIUS95/07241 -99- Hz), 48.41 J=271 Hz), 44.97 J=252 Hz), 44.78 (d, J=252 Hz), 41.85 J=252 Hz), 41.12 J=252 Hz).
Anal. Calcd for C 41
H
47
N
5 0 7
F
2
H
2 0; C, 63.31; H, 6.35; N, 9.00; Found C, 63.37; H, 6.19; N, 8.85.
Example 23 Preparation of N-[4-(N-{2-pv'ridylmethylloxycarbonyl-Lvalyl)amino-2,2-difluoro-1,3-dioxo-5--(4-benzyloxy)phenylpenty. I-o-methyl-D-valinol.
MnQBn 0 NH CFkNH >H4 IfOCH 3 Step
A
Preparation of N-[4-{2-pyridylmethylloxycarbonyl-Lvalyl )amino-2, 2-difluoro-3-hydroxy-l-oxo-5-( 4benzyloxy)phenyl-pentyl ]-O-methyl-D-valinol.
Reaction Scheme step The title compound is prepared in a manner analogous to the coupling procedure described in Example 17, step F from the amine prepared in Example 19, Reaction Scheme A, step and the acid prepared in Example 15, step B, in 55% yield, RfO0.23 (silica gel, ethyl acetate); mass spectrum m/z 685 Calcd for C 36
H
46
N
4 0 7
F
2 0.5 H 2 0; C, 62.32; H, 6.83; N, 8.08; Found C, 62.36; H, 6.64; N, 7.91.
Step B of the final title compound.
Reaction Scheme step (f);The final title compound is prepared in a manner analogous to the Swern Oxidation procedure described in Example 17, step G from the above prepared alcohol, in 44% yield, Rf=0.3 (silica gel, WO 96/02499 PCT/US95/07241 -100dichloromethane/ethyl acetate, mass spectrum m/z 683 1 H NMR (DMSO-d 6 6 9.10-8.92 1H), 8.67-8.51 (m, 2H), 7.95-7.87 1H), 7.64-6.82 (several m, 11H), 5.30- 5.03 5H), 4.10-3.93 1H), 3.93-3.78(m, 1H), 3.60- 3.37 3.35-3.25 (several s, 3H), 3.32-3.10 and 2.87- 2.68 2H), 2.14-1.80 2H), 1.10-0.60 12H); 19
F
NMR (DMSO-d 6 6 (CeFe) 52.80 J=267 Hz), 52.21 J=269 Hz), 50.93 J=269 Hz), 52.21 J=269 Hz), 50.93 (d, J=269 Hz), 49.36 J=267 Hz).
Anal. Calcd for C 36
H
44
N
4 0 7
F
2 0.5 H 2 0; C, 62.51; H, 6.56; N, 8.10; Found C, 62.65; H, 6.50; N, 7.93.
Example 24 Preparation of N-(4-(N-{2-pyridylmethyl}oxycarbonyl-Lvalyl)amino-2,2-difluoro-l,3-dioxo-5-(4-benzyloxy)phenylpentyl]-O-benzyl-D-valinol.
OBn 020 0 NH CF H O0 0o Step A Preparation of N-[4-(N-(2-pyridylmethyl}oxycarbonyl-Lvalyl)amino-2,2-difluoro-3-hydroxy-l-oxo-5-(4- 3 benzyloxy)phenyl-pentyl]-O-benzyl-D-valinol.
Reaction Scheme step The title compound is prepared in a manner analogous to the coupling procedure described in Example 17, step F from the amine prepared in Example 5, step B and the acid prepared in Example 15, step B in 52% yield, Rf=0.28 (silica gel, petroleum ether/ethyl acetate, 3/7).
Step B Preparation of the final title compound.
WO 96/02499 PCT[US95/07241 -101- Reaction Scheme step The final title compound is prepared in a manner analogous to the Swern Oxidation procedure described in Example 17, step G from the above prepared alcohol, in 62% yield, RfO0.
0 7 (Alumina neutral Act III, tetrahydrofuran/dichloromethane/water, 20/10/0.2), Rf=0.27 (silica gel, ethyl acetate/petroleum ether,7/3); mass spectrum m/z 759 1 H NMR (CDCl 3 6 8.73-8.51 7.79-7.65 (mn, 1H), 7.42-6.45 (several mn, 18H), 5.70-5.10 (mn, 4H), 5.06-4.93 (in, 2H), 4.58-4.40 (in, 2H), 4.05-3.80 (in, 2H), 3.74-3.58 and 3.55-3.40 (in, 2H), 3.35- 3.20 and 3.00-2.80 (mn, 2H), 2.13-1.90 (mn, 2H), 1.10-0.50 (mn, 12H) 19 F NMR (CDCl 3 6 (C 6
F
6 50.67 J=273 Hz) 49.50 (broad 48.35 J=273 Hz), 47.24 J=255 Hz), 45.70 J=255 Hz), 42.01 J=255 Hz) 40.00 J=255 Hz).
Anal. Calcd for C 42
H
48
N
4 0 7
F
2 0.5 H 2 0: C, 65.70; H, 6.43; N, 7.30; Found C, 65.69; H, 6.25; N, 7.19.
Example Preparation of N-{4-(N-{3-pyridylmethyl~oxycarbonyl-Lvalyl )ainino-2 ,2-difluoro-1, 3-dioxo-5- C4-benzyloxy )phenylpentyl]-O-[2-(2-methoxyethoxy)-l-ethyl]-D-valinol.
N OBn 00
N
NH CF 2 NH0 H0 .1 0 0 Step A Preparation of N-trityl-O-[2-(2-inethoxyethoxy)-l-ethyll-Dvalinol.
The title compound is prepared in a manner analogous to the alkylation procedure described in Example 17, step B from the compound prepared in Example 17, step A and 2-(2- WO 96/02499 PCT/US95/07241 -102methoxyethoxy)ethyl-l-bromide in 86% yield, Rf=0.74 (silica gel, acetone/petroleum ether, 2/8).
Step B Preparation of O-[2-(2-methoxyethoxy)-l-ethyl]-D-valinol.
A solution of N-trityl-O-[2-(2-methoxyethoxy)-l-ethyl]- D-valinol (1.0 g, 2.28 mmol, prepared above) in dry ether saturated with hydrogen chloride (20 mL) is kept for hours at room temperature. The reaction is concentrated under vacuum and the residue is purified by flash chromatography (silica gel, dichloromethane first to elute the trityl alcohol and then dichloromethane/diethylamine, 95/5, Rf=0.20) to provide the title compound (0.46 g, 100%) as a colorless oil.
Step C Preparation of N-[4-tert-butoxycarbonylamino-2,2-difluoro- 3-hydroxy-l-oxo-5-(4-benzyloxy)phenyl-pentyll-O-[2-(2methoxyethoxy)-l-ethyll-D-valinol.
Reaction Scheme A, step The title compound is prepared in a manner analogous to the procedure described in Example 19, step D from the ester of Example 1, step B and O-[2-(2-methoxyethoxy)-l-ethyl]-D-valinol, prepared above, in 51% yield, Rf=0.37 (silica gel, petroleum ether/ethyl acetate, 3/7).
Step D Preparation of N-[4-amino-2,2-difluoro-3-hydroxy-l-oxo-5- (4-benzyloxy)phenyl-pentyl]-O-[2-(2-methoxyethoxy)-lethyl]-D-valinol.
Reaction Scheme A, step The title compound is prepared in a manner analogous to the deprotection procedure described in Example 17, step C from N-[4-tertbutoxycarbonylamino-2,2-difluoro-3-hydroxy-l-oxo-5-(4benzyloxy)phenyl-pentyl]-0-[2-(2-methoxyethoxy)-1-ethyl]-Dvalinol, prepared above, in 97% yield which is used directly in the next step; mass spectrum m/z 539 WO 96/02499 PCT/US95/07241 -103- Step E Preparation of N-[4-(N-{3-pyridylmethyl}oxycarbonyl-Lvalyl)amino-2,2-difluoro-3-hydroxy-l-oxo-5-(4benzyloxy)phenyl-pentyl]-O-[2-(2-methoxyethoxy)-1-ethyl]-Dvalinol.
Reaction Scheme step The title compound is prepared in a manner analogous to the coupling procedure described in Example 17, step F from the amine prepared above and the acid prepared in Example 8, step A, in 52% yield, Rf=0.18 (silica gel, ethyl acetate); mass spectrum m/z 773 Step F Preparation of final title compound.
Reaction Scheme step The final title compound is prepared in a manner analogous to the Swern Oxidation procedure described in Example 17, step G from the above prepared alcohol, in 76% yield, Rf=0.09 (silica gel, ethyl acetate); mass spectrum m/z 771 1H NMR (DMSO-d 6 6 9.05-8.88 1H), 8.77-8.55 2H), 8.40-8.27 1H), 7.94-7.81 1H), 7.76-6.55 (several m, 11H), 5.31-4.97 5H), 4.10-3.79 2H), 3.76-3.37 10H), 3.32 (s, 3H), 3.29-3.11 and 2.89-2.60 2H), 2.09-1.79 2H), 1.11-0.44 12H); 19 F NMR (DMSO-d 6 6 (C 6
F
6 52.97 (d, J=267 Hz), 52.12 J=269 Hz), 50.88 J=269 Hz), 49.45 J=254 Hz), 45.35 J=267 Hz), 48.17 J=254 Hz), 46.17 J=254 Hz), 44.31 J=254 Hz).
Anal. Calcd for C 40
H
52
N
4 OgF 2 C, 62.32; H, 6.80; N, 7.27; Found C, 61.78; H, 6.77; N, 7.12.
WO 96/02499 PCT/US95/07241 -104- Example 26 Preparation of N-[4-(N-{3-pyridylmethyl)oxycarbonvl-Lvalyl)amino-2,2-difluoro-1,3-dioxo-5-(4-f2-Nmorpholyl]ethyloxy)phenyl-pentyl]-O-methyl-D-valinol.
N 0 -'0 0 NH CF 2
NH
H4
OCH
3 0 ,0 Step A Preparation of N-[ 4 -tert-butoxycarbonylamino-2,2-difluoro- 3-hydroxy-l-oxo-5-(4-hydroxy)phenyl-pentyll-O-methyl-Dvalinol.
Reaction Scheme A, step The title compound is prepared in a manner analogous to the procedure described in Example 17, Step D, from the ester of Example 21, step A and the amine prepared in Example 19, step C, in 83% yield, Rf=0.15 (silica gel, petroleum ether/ethyl acetate, 6/4); mass spectrum m/z 461 Step
B
Preparation of N-[ 4 -tert-butoxycarbonylamino-2,2-difluoro- 3-hydroxy-l-oxo-5-(4-(2-N-morpholyl}ethyloxy)phenylpentyl]-O-methyl-D-valinol.
Reaction Scheme A, step A mixture of N-[4-tertbutoxycarbonylamino-2,2-difluoro-3-hydroxy-l-oxo-5-(4hydroxy)phenyl-pentyl]-O-methyl-D-valinol (1.117 g, 2.43 mmol, prepared above), N-( 2 -chloroethyl)morpholine HC1 (0.633 g, 3.40 mmol), cesium carbonate (2.69 g, 8.26 mmol) and potassium iodide (0.056 g, 0.34 mmol) in dry DMF mL) is stirred for 140 hours at room temperature. The reaction mixture is then diluted with ethyl acetate (100 mL) and washed with brine (2 x 100 mL). The aqueous washes are extracted with ethyl acetate (100 mL). The combined organic extracts are dried over anhydrous sodium sulfate, WO 96/02499 PCT/US95/07241 -105filtered and concentrated under vacuum. The residue is purified by flash chromatography (silica gel, ethyl acetate, Rf=0.18) to provide the title compound (0.993 g, 71%).
Step C Preparation of N-[4-amino-2,2-difluoro-3-hydroxy-l-oxo-5- (4-{2-N-morpholyl}ethyloxy)phenyl-pentyl]-0-methyl-Dvalinol.
Reaction Scheme A, step The title compound is prepared in a manner analogous to the deprotection procedure described in Example 17, step C from N-[4-tertbutoxycarbonylamino-2,2-difluoro-3-hydroxy-l-oxo-5-(4-{2-Nmorpholyl}ethyloxy)phenyl-pentyl]-O-methyl-D-valinol, prepared above, in 85% yield which is used directly in the next step.
Step D Preparation of N-[4-(N-(3-pyridylmethyl}oxycarbonyl-Lvalyl)amino-2,2-difluoro-3-hydroxy-l-oxo-5-(4-{2-Nmorpholyl}ethyloxy)phenyl-pentyl]-O-methyl-D-valinol.
Reaction Scheme step The title compound is prepared in a manner analogous to the procedure described in Example 17, Step F from the amine prepared above and the acid prepared in Example 8, step A, in 80% yield, Rf=0.19 (silica gel, ethyl acetate/methanol, mass spectrum m/z 708 Step E Preparation of final title compound.
Reaction Scheme step A mixture of the above prepared alcohol (0.280 g, 0.396 mmol), 1,1,1-triacetoxyl,l-dihydro-l,2-benziodoxol-3(lH)-one (0.672 g, 1.583 mmol, Dess-Martin periodinane) and tert-butanol (0.075 mL, 0.792 mmol) in freshly distilled dichloromethane (10 mL, distilled over P 2 0 5 is stirred for 15 minutes at room temperature. The reaction mixture is then hydrolyzed with WO 96/02499 PCT/US95/07241 -106isopropanol (1.2 mL) and concentrated under vacuum. The residue is suspended in dichloromethane (3 mL), filtered through a Fluropore filter which is rinsed with dichloromethane (2 x 1 mL). The filtrate is concentrated under vacuum and the residue purified by flash chromotagraphy (2x, silica gel, dichloromethane/methanol, 99/1 to remove the by-products of the Dess-Martin Reagent, then 98/2 and 96/4 to elute the title compound) to provide the title compound (0.112 g, Rf=0.11 (dichloromethane/methanol, 95/5); mass spectrum m/z 706 1 H NMR (DMSO-d 6 6 8.99-8.70 1H), 8.64-8.43 (m, 3H), 7.82-7.73 1H), 7.47-6.70 (several m, 6H), 5.16- 4.92 3H), 4.08-3.95 (broad s, 2H), 3.95-3.71 1H), 3.71-3.65 1H), 3.60-3.48 (broad s, 4H), 3.43-3.34 (m, 2H), 3.22-3.18 (2s, 3H), 3.24-3.03 and 2.77-2.56 2H), 2.54-2.34 4H), 2.02-1.70 2H), 1.10-0.50 12H); 19F NMR (DMSO-d 6 6 (C 6
F
6 52.87 J=267 Hz), 52.22 (d, J=269 Hz), 50.91 J=269 Hz), 49.38 J=267 Hz).
Anal. Calcd for C35H49NSOBF 2 *0.5 H 2 0; C, 58.81; H, 7.05; N, 9.80; Found C, 58.61; H, 6.88; N, 9.93.
WO 96/02499 PCTIUS95/07241 -107- In a further embodiment the present invention provides a method of treating a patient afflicted with a viral infection comprising the administration thereto of an effective antiviral amount of a compound of formula The term "viral infection" as used herein refers to an abnormal state or condition characterized by viral transformation of cells, viral replication and proliferation. Viral infections for which treatment with a compound of formula will be particularly useful include retroviruses such as but not limited to HTLV-I, HTLV-II, HTLV-III (HIV virus), murine leukemia virus, feline leukemia virus, cytomegalovirus(CMV), avian sarcoma virus and the like. In addition treatment with a compound of formula would be useful in treating a wide range of states of HIV infection: AIDS, ARC (AIDS related complex), both symptomatic and asymptomatic, and actual or potential exposure to HIV. For example, the compounds of this invention are useful in preventing infection by HIV after suspected past exposure to HIV by, blood transfusion, accidental needle stick, or exposure to patient blood during surgery.
An "effective antiviral amount" of a compound of formula refers to an amount which is effective, upon single or multiple dose administration to the patient, in controlling the growth of the virus or in prolonging the survivability of the patient beyond that expected in the absence of such treatment. As used herein "controlling a viral infection" refers to slowing, interrupting, arresting or stopping the viral transformation of cells or the replication and proliferation of the virus and does not necessarily indicate a total elimination of the virus.
The present invention further provides a method of inhibiting HIV protease in a patient in need thereof WO 96/02499 PCT/US95/07241 -108comprising administering to said patient an effective inhibitory amount of a compound of formula It is understood that patients suffering from a retrovirus, such as HTLV-III are in need of an HIV protease inhibitor such as a compound of formula As used herein, the term "patient" refers to a warmblooded animal, such as a mammal, which is afflicted with a particular viral infection. It is understood that humans, mice and rats are included within the scope of the term "patient".
Administration of a compound of formula to a patient results in inhibition of HIV protease in the patient. Thus, by treatment of a patient with a compound of formula retroviruses, such as HTLV-III, are inhibited or suppressed.
A patient is in need of treatment with an agent which inhibits HIV protease, such as a compound of formula where the patient is suffering from certain viral infections for which HIV protease is implicated as a contributing factor in the progression of the disease.
Based on standard clinical and laboratory tests and procedures, an attending diagnostician, as a person skilled in the art, can readily identify those patients who are in need of treatment with an agent which inhibits HIV protease, such as a compound of formula An "effective inhibitory amount" of a compound of formula is that amount which is effective, upon single or multiple does administration to a patient, in providing an inhibition of HIV protease.
WO 96/02499 PCT/US95/07241 -109- As used herein the term "effective amount" refers to an effective antiviral or inhibitory amount of a compound of formula An effective amount can be readily determined by the attending diagnostician, as one skilled in the art, by the use of known techniques and by observing results obtained under analogous circumstances. In determining the effective amount or dose, a number of factors are considerec by the attending diagnostician, including, but not limited to: the species of mammal; its size, age, and general health; the specific viral infection involved; the degree of or involvement or the severity of the viral infection; the response of the individual patient; the particular compound administered; the mode of administration; the bioavailability characteristics of the preparation administered; the dose regimen selected; the use of concomitant medication; and other relevant circumstances.
An effective amount of a compound of formula is expected to vary from about 0.1 milligram per kilogram of body weight per day (mg/kg/day) to about 100 mg/kg/day.
Preferred amounts are expected to vary from about 0.5 to about 10 mg/kg/day.
In effecting treatment of a patient afflicted with a viral infection, a compound of formula can be administered in any form or mode which makes the compound bioavailable in effective amounts, including oral and parenteral routes. For example, compounds of formula (I) can be administered orally, subcutaneously, intramuscularly, intravenously, transdermally, intranasally, rectally, and the like. Oral administration is generally preferred. One skilled in the art of preparing formulations can readily select the proper form and mode of administration depending upon the particular characteristics of the compound selected, the viral WO 96/02499 PCT/US95/0724 1 -110infection to be treated, the stage of the infection, and other relevant circumstances.
The compounds of formula can be administered alone or in the form of a pharmaceutical composition in combination with pharmaceutically acceptable carriers or excipients, the proportion and nature of which are determined by the solubility and chemical properties of the compound selected, the chosen route of administration, and standard pharmaceutical practice. The compounds of the invention, while effective themselves, may be formulated and administered in the form of their pharmaceutically acceptable salts for purposes of stability, convenience of crystallization, increased solubility and the like.
In another embodiment, the present invention provides compositions comprising a compound of formula in admixture or otherwise in association with one or more inert carriers. These compositions are useful, for example, as assay standards, as convenient means of making bulk shipments, or as pharmaceutical compositions. An assayable amount of a compound of formula is an amount which is readily measurable by standard assay procedures and techniques as are well known and appreciated by those skilled in the art. Assayable amounts of a compound of formula will generally vary from about 0.001% to about of the composition by weight. Inert carriers can be any material which does not degrade or otherwise covalently react with a compound of formula Examples of suitable inert carriers are water; aqueous buffers, such as those which are generally useful in High Performance Liquid Chromatography (HPLC) analysis; organic solvents, such as acetonitrile, ethyl acetate, hexane and the like; and pharmaceutically acceptable carriers or excipients.
More particularly, the present invention provides pharmaceutical compositions comprising a WO 96/02499 PCT/US95/07241 -111therapeutically effective amount of a compound of formula in admixture or otherwise in association with one or more pharmaceutically acceptable carriers or excipients.
The pharmaceutical compositions are prepared in a manner well known in the pharmaceutical art. The carrier or excipient may be a solid, semi-solid, or liquid material which can serve as a vehicle or medium for the active ingredient. Suitable carriers or excipients are well known in the art. The pharmaceutical composition may be adapted for oral or parenteral use and may be administered to the patient in the form of tablets, capsules, suppositories, solution, suspensions, or the like.
The compounds of the present invention may be administered orally, for example, with an inert diluent or with an edible carrier. They may be enclosed in gelatin capsules or compressed into tablets. For the purpose of oral therapeutic administration, the compounds may be incorporated with excipients and used in the form of tablets, troches, capsules, elixirs, suspensions, syrups, wafers, chewing gums and the like.
These preparations should contain'at least 4% of the compound of the invention, the active ingredient, but may be varied depending upon the particular form and may conveniently be between 4% to about 70% of the weight of the unit. The amount of the compound present in compositions is such that a suitable dosage will be obtained. Preferred compositions and preparations according to the present invention are prepared so that an oral dosage unit form contains between 5.0-300 milligrams of a compound of the invention.
The tablets, pills, capsules, troches and the like may also contain one or more of the following WO 96/02499 PCT/US95/07241 -112adjuvants: binders such as microcrystalline cellulose, gum tragacanth or gelatin; excipients such as starch or lactose, disintegrating agents such as alginic acid, Primogel, corn starch and the like; lubricants such as magnesium stearate or Sterotex; glidants such as colloidal silicon dioxide; and sweetening agents such as sucrose or saccharin may be added or a flavoring agent such as peppermint, methyl salicylate or orange flavoring. When the dosage unit form is a capsule, it may contain, in addition to materials of the above type, a liquid carrier such as polyethylene glycol or a fatty oil. Other dosage unit forms may contain other various materials which modify the physical form of the dosage unit, for example, as coatings. Thus, tablets or pills may be coated with sugar, shellac, or other enteric coating agents. A syrup may contain, in addition to the present compounds, sucrose as a sweetening agent and certain preservatives, dyes and colorings and flavors. Materials used in preparing these various compositions should be pharmaceutically pure and non-toxic in the amounts used.
For the purpose of parenteral therapeutic administration, the compounds of the present invention may be incorporated into a solution or suspension.
These preparations should contain at least 0.1% of a compound of the invention, but may be varied to be between 0.1 and about 50% of the weight thereof. The amount of the inventive compound present in such compositions is such that a suitable dosage will be obtained. Preferred compositions and preparations according to the present invention are prepared so that a parenteral dosage unit contains between 5.0 to 100 milligrams of the compound of the invention.
The solutions or suspensions may also include the one or more of the following adjuvants: sterile WO 96/02499 PCT/US95/07241 -113diluents such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl paraben; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylene diaminetetraacetic acid; buffers such as acetates, citrates or phosphates and agents for the adjustment of tonicity such as sodium chloride or dextrose. The parenteral preparation can be enclosed in ampules, disposable syringes or multiple dose vials made of glass or plastic.
The present invention is also directed to combinations of the HIV protease-inhibitory compounds with one or more agents useful in the treatment of AIDS, such as, for example, with known antiviral agents suitable for treating HIV 1 and HIV 2 viral infections, AZT, with or without a PNPase inhibitor, or in conjunctive therapy with DDI and a PNPase inhibitor.
WO 96/02499 PCT/US95/07241 -114- The compounds of this invention may be assayed fcr their HIV-protease inhibition using the following published techniques.
Preparation of Retroviral Enzyme and Assay for Inhibition of the Protease A) Preparation of Retroviral Enzyme To prepare the recombinant protease, the HIV protease is expressed via E.Coli by the published work of C. Guinet, et al., in European Journal of Pharmacology, Molecular Pharmacology Section, 172 (1989) 443-451.
B) Assay for Inhibition of Recombinant Viral Protease Inhibition of the reaction of the protease with a peptide substrate [Ser-Gln-Asn-Tyr-Pro-Ile-Val-NH 2 Km 1 mM] are in 50 mM Na acetate, 10% glycerol, ethyleneglycol, pH 5.5, at 37 0 C for 1 hour. Various concentrations of inhibitor in 10 pl DMSO are added to l of assay solution and the reaction is initiated by the addition of 10 Ul (1.6 Ug) of recombinant protease. The reaction is quenched with 16 l1 of 4 M perchloric acid. Products of the reaction are separated by HPLC (VYDAC wide pore 5 cm C-18 reverse phase, acetonit-ile gradient, 0.1% trifluoroacetic acid). The extent of inhibition of the reaction is determined from the peak heights of the products. HPLC of the products, independently synthesized, provide quantitation standards and confirmation of the product composition.
By following the techniques referenced above, as well as by utilization of other known techniques, as well as by comparison with compounds known to be useful for treatment of the above-mentioned disease states, it is believed that adequate material is available to enable one of ordinary skill in the art to practice the invention.
WO 96/02499 PCT/US95/07241 -115- As is true for most classes of compounds found to be useful in the pharmceutical industry, certain subgeneric groups and certain specific compounds are more preferred such as those exemplified and shown in the following charts.
te 0 4* P2I P 1 6 2-pyr idylmethyloxy 3-pyr idylmethyloxy 2-pyr idylmetthyloxy 3-pyr idylmethyloxy isopropyl isopropyl isopropyl isopropyl (2-pyridyl)methyloxyj benz yl 4-i (3-pyridyl)methyloxy] benzyl 4- (N-morpholyl ehyloxy] benzyl 4- (N-morpholyl )ethyloxy] benzyl 00 oo" DC O
I
Xx11 1r P 1 IR=H9R
I
3-pyridylmethyloxy 3-pyr idylmethyloxy 0 0 I
N
0 3-pyridylmethyloxy t-butyl isopropyl isopropyl isopropyl 4- (N-morpholyl )ethyloxy] benzyl 4- (N-morpholyl )ethyloxy I benzyl D- 0 N OCH 3 0 0ci D 0 3
DN
4- (N-morpholyl )ethyloxy I benzyl 2-(N-piperidyl)ethyloxy] benzyl S. S.
S 55 S *S S S. 5 S 5 S S 5 S *9~ S 5 S. S S 5 5 S 55 5* 55 55 *5 .S se 1 2 1 P1 R 5 =l R16 3-pyridylmethyloxy 2-pyr idylmethyloxy 2-'pyr idyimetliyloxy 2-pyr idylmethyloxy isopropyl isopropyl L-butyl isopropyl benzyl 4- (benzyloxy )benzyl 4-f (3-pyridyl)methyloxyl benzyl 4-f (3-pyridyl)methyioxyl benzyl 0
H~~I
0 C7 t) 0\/
NJ
D- 0 N/\H
S
S 4* S S *t S S S **5 S S S *5 S S S S. *S R 5= ,9R 1- I -t 0- 0 0s isopropyl i so prop yl.
isopropyl isopropyl 0 00 4-f 2-(N-morpholyl) ethyloxy Ibenzyl 4-f (3-pyridyl)methyloxy Ibenzyl (3-pyridyl) methyloxy Jbenzyl 2-f N-rorpholyl) ethyloxylIbenzyl 0 0 D N CO it 0 C. C. C.
C Ce be C. C. C C *t C CC C Ce C CC. *.Ce C C C C C C C CCC C C C C be CC *e R, 1 I '2 1 P 1 I R 5 R16 I N
N
0 3-pyridylmethyloxy 2-pyr idylmethyloxy isopropyl isopropyl isopropyl isopropyl 4-f 2-(N-morpholyl) ethyloxylIbenzyl 4-f 2-(N-piperidyl) ethyloxy Ibenzyl 4-f 2-(N-piperidyl) e thy lttxy I ben zyl 4- (N-morpholyl) ethyloxy ]benz~'i 0 D N 0 0
D
0
ON
.9 9. 9. 9. 9. 9* .9 99 *9 9 9 .9 9. 9 9 9* 9 9 9 9*99 9 9 9 9 *99 9**9 9 .9 9 9 9 9 9 9 .9 99 99 9999 *9 999 9* Al 1 k' 2 J pi R 5~=H R 2-pyridylmeLhyloxy 0 N 0 00 0 0 0 0 r SO 2 isopropyl isopropyl L-butyl isopropyl 4-f 2-(N-morpholyl) ethyloxy Jbenzyl 4- (benzyloxy )benzyl 4-f 2-(N-piperidyl) ethyloxy Ibenzyl 4-f 2-(N-piperidyl) ethyloxy Ibenzyl 0 0 0 0
ND
00 WO 96/02499 WO 9602499PCT/US95/07241I -122- The following list illustrates compounds according to the present invention; [[(hydroxy)phenylacetyllamino]-3-methyl-.-oxobut-yljamino..
R-oxo-4- (phenylmethoxy (phenylmethy))-benzenepentanamide; 2. 9R*, 13*144-ifur--l methylethyl)-3,5,8,l l-tetraoxo-1,13-diphenyl-6-[ [4- (phenylmethoxy)phenyllmethyl]-12-oxa-2,7,lotriazatetradecan-14-oic acid, methyl ester; phenyl-2-propenyl)aminolbutyllamino]-B-oxo-4- (phenylmethoxy (phenylmethyl )-benz ene-pentanamide; 4. a,a-difluoro-y-[ [2-[2-hydroxy-l-oxo-4- (phenylbutyl)amino]-3-methyl-l-oxobutyl]amino]-B-oxo-4- (phenylmethoxy) (phenylmethyl )-benzene-pentanamide; 3,3-difluoro-4-[ [2-methyl-i- [(phenylmethoxy)methyl]-propyl]aminol-2,4-dioxo-l-[ [4- (phenylmethoxy )phenyl 1-methyl] butyllIamino] carbonyl 1-2methylpropyi ]-S-oxo-4-morpholinepropanamide; [f (hydroxy)phenylacetyl]-amino]-3-methyl-l-oxobutyl]amino]- N-[2-methyl-1-[ (phenylmethoxy)methyljpropyl]-3-oxo-4- (phenylmethoxy)-benzenepentanamide; 7. N-fl-f[ [3,3-difluoro-4-[ [2-methyl-l- [(phenylmethoxy)-methyl]-propyllamino]-2,4-dioxo-l-[ [4- (phenylmethoxy)phenyl]- methyl]-butyl]amino]carbonyl]-2methylpropyl]-6-(4-morpho2.inyl-carbonyl)-3pyridinecarboxamide; WO 96/02499 WO 9602499PCTIUS95/0724 I -12 3- 8. 7,7-difluoro-4,12-bis(l-methylethyl)-6,8,1l-trioxol-phenyl-9-[ [4-(phenylmethoxy)phenyllmethyl]-2-oxa-5,1O,13triazatetradecan-14-oic acid, 3-pyridinylmethyl ester; 9. [l-l*2*Iaadfuroy[3mty--[-4 morpholinylsulfonyl)benzoyllamino]-l-oxobutyl]amino]-N-[2methyl-l-[ (phenylmethoxy)methyl ]propyl ]-B-oxo-4- (phenyirnethoxy) -benzenepentanamide; 10. [l-l*2*Iaadfuroy[3mty--[-4 morphol inylcarbonyl )benzoyl ]amino] -1-oxobutyl 1amino] [2methyl-i-[(phenylmethoxy )methyl Ipropyl I-s-oxo-4- (phenylmethoxy) -benz enepentanamide; 11. [[3,3-difluoro-4-[ [2-methyl-i- (phenylmethoxy)methyll-propyllamino]-2,4-dioxo-l-E [4- (phenylmethoxy )phenyl 1-methyl 1butyl Iamino] carbonyl 1-2methyipropyl I-4-morpholineacetamide; 12. [[3,3-difluoro-4-[ [2-methyl-i- [(phenylmethoxy)methyl]-propyllamino]-2,4-dioxo-1-[ [4- (phenylmethoxy)phenyllmethyl]-butyllaminolcarbonyll-2methylpropyl I-lH-imidazole-l-acetamide; 13. [l-[[[3,3-difluoro-2,4-dioxo-1-V[4- (phenylmethoxy)phenyl 1-methyl (2pyridinylmethyl)aminolbutyllaminolcarbonyl]- 2methylpropyll-carbamic acid, 3-pyridinylmethyl ester; WO 96/02499 WO 9602499PCTfUS95/0724 1 -124- 14. [1-[[[3,3-difluoro-2,4-dioxo-1-[[4- (phenylmethoxy )phenyl 1-methyl 1-4- pyridinylmethyl )amino] butyllIamino] carbonyl 1-2methyilpropyl]-carbamic acid, 3-pyridinylmethyl ester; [[3,3-difluoro-2,4-dioxo-l-[ [4-(phenylmethoxy)phenyllj-methyl]-4-[ (2-pyridinylmethyl)aminolbutyl]aminolcarbonyll- 2-methylpropyll-carbamic acid, 2pyridinylmethyl ester; 16. [[3,3-difluoro-2,4-dioxo-1-[[14-(phenylmethoxy)phenyl]-methyl]-4-[ (3-pyridinylmethyl)amino]butyl]aminolcarbonyl)- 2-methylpropyll-carbamic acid, 2pyridinylmethyl ester; 17. N-[4-(N-{2-pyridylmethyl~oxycarbonyl-L-valyl)amino- 2 2 -difluoro-1,3-dioxo-5-(4-benzyoxy)pheny-penty..a..(3pyridylmethyl )-D-valinol; 18. N-14-(N-{3-pyridylmethylloxycarbonyl-L-valyl)amino- 2 2 -difluoro-1,3-dioxo-5-(4-benzyloxy)phenyl-pentyl]-i-(3pyr idylmethyl )-D-valinol; 19. N-[4-(N-{3-pyridylmethyl~oxycarbonyl-L-valyl)amino- 2 2 -difluoro-1,3-dioxo-5-(4-benzyloxy)phenyl-pentyll-0methyl-D-valinol; N-t4-(N-(3-pyridylmethylloxycarbonyl-L-valyl)amino- 2, 2-difluoro-1, 3-dioxo-5- 4 -benzyloxy)phenyl-pentyl1-0- (2pyridylmethyl)-D-valinol; 21. N-[l4-(N-{2-pyridylmethyl~oxycarbonyl-Lvalyl amino-2, 2-difluoro-1, 3-dioxo-5- 3pyridylmethyl~oxy)phenyl-pentyl] (3-pyridylmethyl valinol; WO 96/02499 WO 9602499PCT1US95/0724 1 -12 22. N-(4-(N-{2-pyridylmethyl~oxycarbonylLvalyl)amino- 2, 2-difluoro-1 ,3-dioxo-5- (4-benzyloxy )phenyl-pentyl pyridylmethyl )-D-valinol; 23. N-114-(N-{2-pyridylmethyl)oxycarbonylL.
valyl)amino- 2 ,2-difuoro-1,3doxo5..(4-benzyloxy)phenyl.
pentyl ]-O-methyl-D-valinol; 24. N-{4-(N-{2-pyridylmethyl}oxycarbonyl-L-valyl)amino- 2 2 -difluoro-1,3-dioxo-5-(4-benzyloxy)phenyl-pentyl1>o benzyl-D-valinol; N-{4-(N-{3-pyridylmethyl~oxycarbonyl..L-valyl)amino- 2 2 -difluoro-1,3-dioxo-5-(4benzyoxy)pheny-pentyl]-Or2- 2 -methoxyethoxy)-l-ethyl-D.valinol; and 26. N-[4-(N-{3-pyridylmethylloxycarbonyl.L-valyl)amino- 2 2 -difluoro-1,3-dioxo-5-(4-{2-N-morpholylIethyloxy)phenyl1 pentyl]I-O-methyl-D-valinol.
Claims (35)
1. A compound of the formula: P 2 P 1 1 I R1--CNH-CH--CNH-CH--C-CF 2 -C--NR 5 R 6 II II II II O O O O b0 0 x and the stereoisomers, hydrates, isosteres and the pharmaceutically acceptable salts thereof wherein P 1 is T C1-6 alkylene wherein T is and T' is or hydrogen, wherein each of W and W' are independently C 1 6 alkylene or nothing, provided that W is C2- 6 alkylene when W is directly attached to a nitrogen atom in R, provided that W' is C2- 6 alkylene when W' is directly attached to a nitrogen atom in R', provided that W or W' are each independently C 1 -6 alkylene when R or R' are each independently an aryl; P 2 is C 1 6 alkyl, cyclopentyl, hydroxy C,-6 alkyl, phenyl, benzyl or 3-tetrahydrofuryl; WO 96/02499 PTU9/74 PCTIUS95/07241 -127- R and RI are each independently -CH 2 CHO, hydroxy .C 1 6 alkyl, C 1 6 alkoxy C 1 6 alkyl, C 1 6 alkyl, 10 (R3)d C 1 6 alkenyJlene, piperazinyl, substituted piperazinyl, piperidyl, morphoJlinyl, pyridyl, pyrazinyl, pyrimidinyl or phenyl wherein substituted piperazinyl is piperazinyl substituted on one nitrogen atom thereof with CHO, C(O)NHR' 4 C 1 4 alkyl or C0 2 R 4 -128- R, is N ~a (CH 2 sI"- (CH 2 )0- 0 a. 04 a a a a a a a a a a. a a a.. a a a a a a a a a. a. a. a a N OH -MV H)-CH- O H CH- 0 0- N-CCH 2 __CH2- or Risc2- 6 alkenyl, Cl-r, alkoxy C 1 6 alkylf hydroxy C 1 6 alkyi, C 1 6 alkyl or OH; R 4 is C 1 6 alkyl, phenyl or benzyl; R' 4 is hydrogen or C 1 alkyl; WO 96/02499 WO 9602499PCTIUS95/07241 -129- R 5 is hydrogen, CI- 1 5 alkyl, OH hydroxy Cl.- 15 alkyl, Cj- 6 alkoxy, -CH([(CH 2 )d-O-CH2]x-R'8)2, -CH 2 Si(CH 3 2 (R 3 PDL, -(Cl- 6 alkylene)-0R4, Cl..6 alkylenee (HO- (b) CH2 I (d) wherein PDL is -(CH2)a2, 3- or 4-pyridyl, or p- substituted benzyloxy, wherein the substitution is with a nitro, OH, amino, C 1 6 alkoxy, hydroxy C 1 6 alkylene, or halogen; Y is Cl..15 alkyl, hydroxy CI- 1 5 alkyl, C1- 6 alkyl or -(CH2)e-C6H4-(V)e'; Z is -(CH2)d-O-CHO, C 1 6 alkylene-O-(CH 2 )d-(O-CH2-CH2)e-O-Cl-6 alkyl, CHO, C0 2 R 4 CO 2 NHR 4 -(CH2)d-O-iCH2)d'-R'7, -(CH2)e-OR4 or M (CH2)e wherein V is OR 4 or hydroxy C 1 6 alkylene; provided that d'=2 when R'7 is piperazinyl, substituted piperazinyl, piperidyl or morpholinyl; WO 96/02499 PCT/US95/07241 -130- R 6 is as defined for R 5 with the proviso that R 6 is other than hydrogen when R 5 is hydrogen, or R 5 and R 6 are taken together with the nitrogen atom to which they are attached are selected from the group consisting of; -N -N 0 (h) (CH: ,N R3 R3-Si -N -CHO (I) or -N R (n) (m) R 7 is CH 2 0R 4 C(O)NHR 4 or CHO; R' 7 is piperazinyl, substituted piperazinyl, piperidyl, morpholinyl, pyridyl, pyrazinyl, pyrimidinyl or phenyl, wherein substituted piperazinyl is piperazinyl substituted on one nitrogen atom thereof with CHO, C(O)NHR 4 CI- 4 alkyl or C0 2 R 4 R 8 is OH) or =0; R' 8 is pyrimidyl, pyridyl, pyrazinyl or phenyl; WO 96/02499 PCT/US95/07241 -131- a is zero, 1, 2 or 3; b and b' are each independently zero or 1; d and d' are each independently 1 or 2; e and e' are each independently zero, 1 or 2; and x is zero or one.
2. A compound according to claim 1 wherein P 1 is 0--CH 2 O--CH 2
3. A compound according to claim 2 wherein x is one and P 2 is isopropyl.
4. A compound according to claim 3 wherein R 1 is -CH 2 -O- N
5. A compound according to claim 3 wherein R 1 is CH 2 -O- N==
6. A compound according to claim 3 wherein R 5 is hydrogen and R 6 is CH-CH 2 -0-CH 2 CH 3 CH 3 WO 96/02499 WO 9602499PCTIUS95/07241 -132-
7. A compound according to claim 3 wherein R 5 is hydrogen and R 6 is -CH-CH 2 -O-CH 2 CH3H
8. A compound according to claim 3 wherein 1 5 is hydrogen and R 6 is -CH-CH 2 -O-CH 2 C H
9. A compound according to claim 3 wherein R 5 is hydrogen and R 6 is -CH-CH 2 -O -CH 3 CH&?~H 3 A compound according to claim 1 wherein the compound is c,c-difluoro-y-[[(2-(R)- [[(hydroxy)phenylacetyllamino]-3-methyl-l-oxobutyllaminol- a-oxo-4- (phenylmethoxy) (phenylmethyl )-benzene- pentanamide.
11. A compound according to claim 1 wherein the compound is 9R*, 13S*)]-4,4,-difluoro-9-(l- methylethyl)-3,5,8,11-tetraoxo-1,13-diphenyl-6-[ (4- (phenylmethoxy)phenyl Imethyl -12-oxa-2, 7,10- triazatetradecan-14-oic acid, methyl ester. WO 96/02499 WO 96/ 2499PCT/US95/0724 1 -133-
12. A compound according to claim 1 wherein the compound is a,a-difluoro-y-[I[3-methyl-l--oxo-2-[ (-oxo- trans-3-phenyl-2-propenyl)amino]butyl Jamino]-6-oxo-4- (phenylmethoxy) (phenylmethyl )-benzene--pentanamide.
13. A compound according to claim 1 wherein the compound is ca,c-difluoro-y-[ [2-[2-hydroxy-l-oxo-4- (phenylbutyl )amino] -3-methyl--l-oxobutyll]amino I-B-oxo-4- (phenylmethoxy)-N-(phenylmethyl)-benzene-pentalamide.
14. A compound according to claim 1 wherein the compound is [[3,3-difluoro-4-[ [2-methyl-i- [(phenylmethoxy)methylil-propyllaminoil-2,4-dioxo-l-[ [4- (phenylmethoxy)phenyl]-methyllbutyllaminolcarbonyl]-2- methyipropyl ]-B-oxo-4-morpholinepropanamide. A compound according to claim 1 wherein the compound is I-a,ct-difluoro-y-[ (hydroxy)phenylacetyl ]-amino]-3-methyl-1-oxobutyl)amifl> N-[2-methyl-l-t (phenylmethoxy)methylllpropyl]-B-oxo-4- (phenylmethoxy) -benzenepentanamide.
16. A compound according to claim 1 wherein the compound is [3,3-difluoro-4-[ [2-methyl-i- [(phenylmethoxy)-methyl]-propyllamino]-2,4-dioxo-l-[ [4- (phenylmethoxy)phenyll- methyll-butyllaminolcarbonyl methyipropyl (4-morpholinyl-carbonyl pyridinecarboxamide.
17. A compound according to claim 1 wherein the compound is 7,7-difluoro-4,12-bis(l-methylethyl)-6,8,11- trioxo-1-phenyl-9-[ [4-(phenylmethoxy)phenyllmethyl]-2-oxa- 5,10,13-triazatetradecan-14-oic acid, 3-pyridinylmethyl ester. WO 96/02499 PCTJUS95/0724 1 -13 4-
18. A compound according to claim 1 wherein the compound is ]-c,c-difluoro--y-[ [3-methyl-2-[[4- (4-morpholinylsulfonyl )benzoyl Iamino] -1-oxobutyl ]amino] -N- [2-methyl--i(phenylmethoxy)methyllpropy1]-B-oxo-4- (phenylmethoxy) -benzenepentanamide.
19. A compound according to claim 1 wherein the compound is ]-a,a-difluoro-y-II[3-methyl-2-[ [4- 4 -morpholinylcarbonyl)benzoyllamino]lloxobutyl~amino>.N- [2-methyl-1-[ (phenylmethoxy)methyllpropylBoxo4- (phenylmethoxy) -benz enepentanamide. A compound according to claim 1 wherein the compound is [[3,3-difluoro-4-[ [2-methyl-i- (phenylmethoxy)methyl-propyllaminol2,4dioxol[ [4- (phenylmethoxy )phenyl I-methyl Ibutyl 71amino] carbonyl 1-2- methyipropyl I- 4 -morpholineacetamide.
21. A compound according to claim 1 wherein the compound is [[3,3-difluoro-4-[ [2-methyl-i- (phenylmethoxy)methy-propylamino24dioxo-l-[[4- (phenylmethoxy)phenyllmethylj-butyllaminolcarbonyl>-2- methyipropyl ]-lH-imidazole-1-acetamide.
22. A compound according to claim 1 wherein the compound is [[3,3-difluoro-2,4-dioxo-l-[ [4- (phenylmethoxy )phenyl ]-methyl 1-4- pyridinylmethyl)aminolbutyl~aminolcarbonyl]- 2- methyipropyll-carbamic acid, 3-pyridinylmethyl ester.
23. A compound according to claim 1 wherein the compound is [1-i[[[3,3-difluoro-2,4-dioxo-l-[ [4- (phenylmethoxy )phenyl 1-methyl 1-4- pyridinylmethyl)aminolbutyljamino]carbnyl2- methyipropyll-carbamic acid, 3-pyridinylmethyl ester. WO 96/02499 PCTJUS95/0724 1 -135-
24. A compound according to claim 1 wherein the compound is fl-f[ f 3 ,3-difluoro-2,4-dioxo-l-f f4- (phenylmethoxy) -phenyl ]-methyl 1-4- pyridinylmethyl)aminolbutyl]-aminolcarbonyl>. 2- methyipropyll-carbamic acid, 2 -pyridinylmethyl ester. A compound according to claim 1 wherein the compound is fl-[[f[ 3 ,3-difluoro-2,4-dioxo-l-f [4- (phenylmethoxy) -phenyl 1-methyl] pyridinylmethyl)aminolbutyl]-aminolcarbonyl1> 2- methyipropyl 1-carbamic acid, 2 -pyridinylmethyl ester.
26. A compound according to claim 1 wherein the compound is 4-(N-{2-pyridylmethyl~oxycarbonyl-L valyl)amino-2,2-difluoro-1l3-.dioxo-.5.(4benzyoxy)phenyl pentyl 1-0- (3-pyr idylmethyl )-D-valinol.
27. A compound according to claim 1 wherein the compound is N-f 4-(N-(3-pyridylmethyl}oxycarbonylL- valyl)amino-2,2-difluoro-1,3-dioxo5(4-benzyloxy)phenyl- pentyl 1-0- (3-pyr idylmethyl )-D-valinol.
28. A compound according to claim 1 wherein the compound is N-f 4-(N-{3-pyridylmethyl~oxycarbonylL valyl)amino-2,2-difluoro1,3-dioxo5(4benzyoxy)phenyl. pentyl ]-0-methyl-D-valinol.
29. A compound according to claim 1 wherein the compound is N-{4-(N-{3-pyridylmethyl~oxycarbony.L- valyl)amino- 2 2 -difluoro1,3-dioxo5(4.benzyloxy)phenyl. pentyl 1-0- (2-pyridylmethyl )-D-valinol. A compound according to claim 1 wherein the compound is N-f4-(N-{2-pyridylmethyl~oxycarbonyl.L- valyl)amino-2,2-difluoro-,3dioxo--( 4 3 pyridylmethyl)oxy)phenyl-pentyl 1-0- (3-pyridylmethyl valinol. WO 96/02499 PCT/US95/07241 -137-
38. A method of inhibiting HIV protease in a patient in need thereof comprising administering to said patient an effective inhibitory amount of a compound according to claim 1.
39. A pharmaceutical composition comprising an assayable amount of a compound of claim 1 in admixture or otherwise in association with an inert carrier.
40. A pharmaceutical composition according to claim 39 for the treatment of a viral infection.
41. A compound according to claim 1 for use in the treatment of a viral infection.
42. A compound according to claim 1 for use in inhibiting HIV protease.
43. Use of a compound of claim 1, optionally in combination with a pharmaceutically acceptable carrier, for the preparation of a pharmaceutical composition for the treatment of a viral infection.
44. Use of a compound of claim i, optionally in combination with a pharmaceutically acceptable carrier, for the preparation of an HIV protease inhibitor. WO 96/02499 WO 9602499PCTfUS95/0724 1 -136- 31. A compound according to claim 1 wherein the compound is N-[4--(N-{2-pyridylmethyl~oxycarbonyl-L- valyl )amino-2, 2-difluoro-l, 3-dioxo-5- (4-benzyloxy)phenyl- pentyl] (2-pyr idylmethyl) -D-valinol. 32. A compound according to claim 1 wherein the compound is N-[4-(N-{2-pyridylmethyl~oxycarbonyl-L- valyl )amino-2, 2-difluoro-l, 3-dioxo-5-( 4-benzyloxy)phenyl- pentyl I-0-methyl-D-valinol. 33. A compound according to claim 1 wherein the compound is N-{4-(N-(2-pyridylmethylloxycarbonyl-L- valyl )amino-2, 2-difluoro-1, 3-dioxo-5- (4-benzyloxy )phenyl- pentyl )-0-benzyl-D-valinol. 34. A compound according to claim 1 wherein the compound is N-{4-(N-(3-pyridylmethyl)oxycarbonyl-L- valyl )amino-2, 2-difluoro-l, 3-dioxo-5- (4-benzyloxy)phenyl- pentylI-0-[2-(2-methoxyethoxy)-l-ethyl>D-.valinol. A compound according to claim 1 wherein the compound is 4-(N-{3-pyridylmethyl}oxycarbonyl-L- valyl )amino-2, 2-difluoro-1, 3-dioxo-5- (4-f 2-N- morpholyl ]ethyloxy)phenyl-pentyl]-0-methyl-D-valinol. 36. A method of treating a patient suffering from a viral infection comprising administering to said patient an effective antiviral amount of a compound according to claim 1. 37. A method of controlling a viral infection in a patient afflicted therewith comprising administering thereto an effective antiviral amount of a compound according to claim 1. WO 96/02499 PCT/US95/07241 -138- A process for the preparation of a compound of the formula: P 2 P 1 R--CNHCH-CH-NH-H-C-CF 2 -C--NR5R6 II II II II SO O O x and the stereoisomers, hydrates, isosteres and the pharmaceutically acceptable salts thereof wherein Pl is T T' C 1 6 alkylene wherein T is and T' is or hydrogen, wherein each of W and W' are independently C 1 -6 alkylene or nothing, provided that W is C 2 6 alkylene when W is directly attached to a nitrogen atom in R, provided that W' is C 2 -6 alkylene when W' is directly attached to a nitrogen atom in R', provided that W or W' are each independently C1-6 alkylene when R or R' are each independently an aryl; P2 is C 1 6 alkyl, cyclopentyl, hydroxy C_g 6 alkyl, 30 phenyl, benzyl or 3-tetrahydrofuryl; 3O WO 96/02499 WO 9602499PCTIUS95/0724 1 -139- R and R' are each independently -CH 2 CHO, hydroxy C 1 6 alkyl, C 1 6 alkoxy C 1 6 alkyl, C 1 6 alkyl, -G(R3)d C 1 6 alkenylene, piperazinyl, substituted piperazinyl, piperidyl, morpholinyl, pyridyl, pyrazinyl, pyrimidinyl or phenyl wherein substituted piperazinyl is piperazinyl substituted on one nitrogen atom thereof with CHO, C(O)NHR' 4 CI- 4 alkyl or C0 2 R 4 -140- R 1 is N S(CH 2 )O- 0 a a a a a. a. a a a a. a a a a a a a a a a. at a a a a. a a OH (CH 2 2 -CH- N-CH 2 OH 0 I H N-CCH 2 _CH- or N R 3 is C 2 -C 6 alkenyl, C 1 -G 6 alkoxy C I-c6 aly hydroxy C1-. 6 alkyl, C1-. 6 alkyl or OH; R 4 is C 1 6 alkyl, phenyl or benzyl; R' 4 is hydrogen or C1- 6 alkyl; WO 96/02499 PCTJUS95/07241 -141- R 5 is hydrogen, C 1 15 alkyl, OH, hydroxy C 1 15 alkyl, C 1 j 6 alkoxy, -CH([(CH2)d-O-CH 2 ]x-R' 8 2 -CH 2 Si(CH 3 2 (R 3 PDL, -(C 1 6 alkylene)-OR 4 MVe HO C 1 -6 alkylene -K J I (CH2)b (b) or N (d) wherein PDL is 3- or 4-pyridyl, or p- substituted benzyloxy, wherein the substitution is with a nitro, OH, amino, C 1 6 alkoxy, hydroxy C1- 6 alkylene, or halogen; Y is C 1 15 alkyl, hydroxy Cl. 1 5 alkyl, C 1 -6 alkyl or -(CH2)e-C6H4(V)eI; Z is -(CH2)d-O-CHO, C 1 -6 alkylene-O-(CH2)d-(O-CH2-CH 2 )e-O-Cl- 6 alkyl, CHO, CO 2 R 4 C0 2 NHR 4 -(CH2)d-O-(CH2)ds-R' 7 -(CH2)e-OR4 or (CH2)e wherein V is OR 4 or hydroxy C1- 6 alkylene; provided that d'=2 when R'7 is piperazinyl, substituted piperazinyl, piperidyl or morpholinyl; WO 96/02499 PCT/US95/07241 -142- R 6 is as defined for R 5 with the proviso that R 6 is other than hydrogen when R 5 is hydrogen, or R 5 and R 6 are taken together with the nitrogen atom to which they are attached are selected from the group consisting of; -N -N O (h) (CH2) /N R3 I R3-Si I3 I 0 -N -CHO (I) or -NNC R (n) (m) R 7 is CH 2 0R 4 C(O)NHR 4 or CHO; R' 7 is piperazinyl, substituted piperazinyl, piperidyl, morpholinyl, pyridyl, pyrazinyl, pyrimidinyl or phenyl, wherein substituted piperazinyl is piperazinyl substituted on one nitrogen atom thereof with CHO, C(O)NHR 4 C 1 4 alkyl or C0 2 R 4 R 8 is OH) or =0; R' 8 is pyrimidyl, pyridyl, pyrazinyl or phenyl; WO 96/02499 PCT/US95/07241 -143- a is zero, 1, 2 or 3; b and b' are each independently zero or 1; d and d' are each independently 1 or 2; e and e' are each independently zero, 1 or 2; and x is zero or one, comprising oxidizing a compound of formula: P 2 P 1 I I R1--CNH-CH--CNHH-C-C-CF 2 -C-NRsR 6 1 II I II 0 0 OH 0 x wherein all the substituents are defined as above. WO 96/02499 PCT/US95/07241 -144-
46. A process for the preparation of a compound of the formula: P 2 P 1 I I R CNH-CH--CNH-CH-C-CF 2 -C--NRsR6 II II II II 0 0 0 0 -X and the stereoisomers, hydrates, isosteres and the pharmaceutically acceptable salts thereof wherein P 1 is wherein T is and T' is or hydrogen, wherein each of W and W' are independently C 1 6 alkylene or nothing, provided that W is C2- 6 alkylene when W is directly attached to a nitrogen atom in R, provided that W' is C2- 6 alkylene when W' is directly attached to a nitrogen atom in R', provided that W or W' are each independently C 1 6 alkylene when R or R' are each independently an aryl; P 2 is C 1 -6 alkyl, cyclopentyl, hydroxy C 1 6 alkyl, phenyl, benzyl or 3-tetrahydrofuryl; WO 96/02499 WO 9602499PCTUS95/0724 1 -14 R and R' are each independently -CH 2 CHO, hydroxy C 1 6 alkyl, C1- 6 alkoxy C 1 6 alkyl, C 1 -6 alkyl, 0(R3)d Cj.. 6 alkenylene, piperazinyl, substituted piperazinyl, piperidyl, morpholinyl, pyridyl, pyrazinyl, pyrimidinyl or phenyl wherein substituted piperazinyl is piperazinyl substituted on one nitrogen atom thereof with CH0, C(0)NHR' 4 C 1 4 alkyl or C0 2 R 4 -14.6- RI is N (CH 2 )O- (CH 2)0) N s-(CH 2 )O- 0 0e a a a. a a. a a a a. a a a a. a a a p a a a a a. a. a a a a OH -(CH 2 2 L -H- CH=CH- or N OH CH- 0 N -CCH 2 II R 3 is C 2 -C 6 alkenyl, C 1 -C 6 alkoxy Cl-c 6 alkyl, hydroxy CI-6 alkyl, C 1 -6 alkyl or OH; R 4 is C 1 6 alkyl., phenyl or benzyl; R' 4 is hydrogen or CI- 6 alkyl; WO 96/02499 PCTUS95/07241 -147- R 5 is hydrogen, C 1 15 alkyl, OH, hydroxy Cl- 1 5 alkyl, C 1 j 6 alkoxy, -CH([(CH2)d-O-CH 2 2 -CH 2 Si(CH 3 2 (R 3 PDL, -(C 1 6 alkylene)-0R 4 MVe HO C 1 -6 alkylene K I (CH2)b S- (b) N -N or CH2 7 (d) wherein PDL is 3- or 4-pyridyl, or p- substituted benzyloxy, wherein the substitution is with a nitro, OH, amino, C1- 6 alkoxy, hydroxy C1- 6 alkylene, or halogen; Y is Cl. 15 alkyl, hydroxy C 1 1 5 alkyl, C1-6 alkyl or -(CH2)e-C6H4-(V)e'; Z is -(CH2)d-O-CHO, C 1 -6 alkylene-O-(CH2)d-(O-CH 2 -CH 2 )e-O-Cl. 6 alkyl, CHO, C0 2 R 4 CO 2 NHR 4 -(CH2)d-O-(CH2)d-R'7, -(CH2)e-OR 4 or M (CH2)e~K) 7e wherein V is OR 4 or hydroxy C 1 6 alkylene; provided that d'=2 when R'7 is piperazinyl, substituted piperazinyl, piperidyl or morpholinyl; WO 96/02499 PCTIUS95/07241 -148- R 6 is as defined for R 5 with the proviso that R 6 is other than hydrogen when R 5 is hydrogen, or R 5 and R 6 are taken together with the nitrogen atom to which they are attached are selected from the group consisting of; N -N O (h) (CH2)b R3 I (k) R7 R8 N -CHO -N or -No RB (n) R 7 is CH 2 OR 4 C(O)NHR 4 or CHO; R' 7 is piperazinyl, substituted piperazinyl, piperidyl, morpholinyl, pyridyl, pyrazinyl, pyrimidinyl or phenyl, wherein substituted piperazinyl is piperazinyl substituted on one nitrogen atom thereof with CHO, C(O)NHR 4 C1- 4 alkyl or C0 2 R 4 R 8 is OH) or =0; R' 8 is pyrimidyl, pyridyl, pyrazinyl or phenyl; -149- a is zero, 1, 2 or 3; b and b' are each independently zero or 1; d and d' are each independently 1 or 2; e and e' are each independently zero, 1 or 2; and x is zero or one, comprising; a) oxidizing a compound of formula: P2 P 1 R' -CNH-CH CNH-CH-C-CF 2 -C--NR 5 R 6 II II I II 0 0 OH 0 x wherein R' 1 is a protected R 1 group and the remaining substituents are defined as above; b) subsequently deprotecting the oxidized product of step a.
47. A compound according to any of claims 1 to when produced by a process according to any one of claims 45 or 46.
48. A compound according to claim 1 substantially "as hereinbefore defined with reference to any of the examples. 2• 25 49. A method according to any one of claims 36, 37 or 38 substantially as hereinbefore defined. DATED: 15 September, 1997 PHILLIPS ORMONDE FITZPATRICK Attorneys for: MERRELL PHARMACEUTICALS INC.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US27622394A | 1994-07-15 | 1994-07-15 | |
| US08/276223 | 1994-07-15 | ||
| PCT/US1995/007241 WO1996002499A1 (en) | 1994-07-15 | 1995-06-06 | Difluorostatone antiviral agents |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| AU2818995A AU2818995A (en) | 1996-02-16 |
| AU711035B2 true AU711035B2 (en) | 1999-10-07 |
Family
ID=23055718
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| AU28189/95A Ceased AU711035B2 (en) | 1994-07-15 | 1995-06-06 | Difluorostatone antiviral agents |
Country Status (13)
| Country | Link |
|---|---|
| EP (1) | EP0773926A1 (en) |
| JP (1) | JPH10505582A (en) |
| CN (1) | CN1152910A (en) |
| AU (1) | AU711035B2 (en) |
| CA (1) | CA2195125C (en) |
| FI (1) | FI970151A0 (en) |
| HU (1) | HUT76648A (en) |
| IL (1) | IL114558A0 (en) |
| MX (1) | MX9700414A (en) |
| NO (1) | NO970158L (en) |
| NZ (1) | NZ288836A (en) |
| WO (1) | WO1996002499A1 (en) |
| ZA (1) | ZA955722B (en) |
Families Citing this family (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5717093A (en) * | 1993-07-08 | 1998-02-10 | Merrell Pharmaceuticals Inc. | Difluoro statone analogs |
| EP0717731B1 (en) * | 1993-09-09 | 1999-04-21 | Merrell Pharmaceuticals Inc. | Difluoro statone antiviral analogs |
| WO1995021186A1 (en) * | 1994-02-04 | 1995-08-10 | Merrell Pharmaceuticals Inc. | Macrocyclic difluorostatone derivatives useful as antiviral agents |
| US6114380A (en) * | 1995-12-18 | 2000-09-05 | Merrell Pharmaceuticals Inc. | Difluoro statone analogs |
| JP2019513698A (en) | 2016-03-04 | 2019-05-30 | エフ．ホフマン−ラ ロシュ アーゲーＦ． Ｈｏｆｆｍａｎｎ−Ｌａ Ｒｏｃｈｅ Ａｋｔｉｅｎｇｅｓｅｌｌｓｃｈａｆｔ | Novel trifluoromethylpropanamide derivatives as HTRA1 inhibitors |
| JP2019507176A (en) | 2016-03-04 | 2019-03-14 | エフ．ホフマン−ラ ロシュ アーゲーＦ． Ｈｏｆｆｍａｎｎ−Ｌａ Ｒｏｃｈｅ Ａｋｔｉｅｎｇｅｓｅｌｌｓｃｈａｆｔ | Novel difluoroketamide derivatives as HTRA1 inhibitors |
| EP3484859B1 (en) * | 2016-07-18 | 2021-09-15 | F. Hoffmann-La Roche AG | New difluoroketamide derivatives as htra1 inhibitors |
| CN109563084A (en) | 2016-08-23 | 2019-04-02 | 豪夫迈·罗氏有限公司 | Novel trifluoromethyl propanamide derivative as HTRA1 inhibitor |
| EP3504193A1 (en) | 2016-08-23 | 2019-07-03 | H. Hoffnabb-La Roche Ag | New difluoroketamide derivatives as htra1 inhibitors |
| CN109232301B (en) * | 2018-10-15 | 2021-01-01 | 天津希恩思生化科技有限公司 | Preparation method of tetraisopropyl hydrazine |
| GB201819125D0 (en) * | 2018-11-23 | 2019-01-09 | Univ Oxford Innovation Ltd | Biomarkers and uses thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1992012123A1 (en) * | 1991-01-02 | 1992-07-23 | Merrell Dow Pharmaceuticals Inc. | Anti-viral compounds |
-
1995
- 1995-06-06 NZ NZ288836A patent/NZ288836A/en unknown
- 1995-06-06 CA CA002195125A patent/CA2195125C/en not_active Expired - Fee Related
- 1995-06-06 JP JP8505010A patent/JPH10505582A/en not_active Ceased
- 1995-06-06 HU HU9700116A patent/HUT76648A/en unknown
- 1995-06-06 MX MX9700414A patent/MX9700414A/en unknown
- 1995-06-06 EP EP95923745A patent/EP0773926A1/en not_active Withdrawn
- 1995-06-06 AU AU28189/95A patent/AU711035B2/en not_active Ceased
- 1995-06-06 WO PCT/US1995/007241 patent/WO1996002499A1/en not_active Application Discontinuation
- 1995-06-06 CN CN95194114A patent/CN1152910A/en active Pending
- 1995-07-10 ZA ZA955722A patent/ZA955722B/en unknown
- 1995-07-12 IL IL11455895A patent/IL114558A0/en unknown
-
1997
- 1997-01-14 FI FI970151A patent/FI970151A0/en unknown
- 1997-01-14 NO NO970158A patent/NO970158L/en unknown
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1992012123A1 (en) * | 1991-01-02 | 1992-07-23 | Merrell Dow Pharmaceuticals Inc. | Anti-viral compounds |
Also Published As
| Publication number | Publication date |
|---|---|
| NZ288836A (en) | 1998-01-26 |
| CA2195125A1 (en) | 1996-02-01 |
| JPH10505582A (en) | 1998-06-02 |
| CN1152910A (en) | 1997-06-25 |
| NO970158D0 (en) | 1997-01-14 |
| MX9700414A (en) | 1998-05-31 |
| FI970151A (en) | 1997-01-14 |
| ZA955722B (en) | 1996-02-26 |
| EP0773926A1 (en) | 1997-05-21 |
| CA2195125C (en) | 2003-12-16 |
| HUT76648A (en) | 1997-10-28 |
| FI970151A0 (en) | 1997-01-14 |
| AU2818995A (en) | 1996-02-16 |
| WO1996002499A1 (en) | 1996-02-01 |
| NO970158L (en) | 1997-03-14 |
| IL114558A0 (en) | 1995-11-27 |
| HU9700116D0 (en) | 1997-02-28 |
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