HRP960260A2 - Hover boat - Google Patents

Description

The invention relates to certain compounds and their pharmaceutically acceptable salts, which inhibit matrix metalloproteases (MMP), especially MMP-3, MMP-12 and MMP-13. These compounds are therefore useful in the treatment of mammals in conditions that can be alleviated by inhibiting MMPs, particularly MMP-3, MMP-12 and MMP-13.

MMPs form a family, structurally similar to zinc-containing metalloproteases, which participate in the restructuring, restoration and degradation of extracellular matrix proteins as part of normal physiological processes and in pathological conditions. Because they possess a strong destructive potential, MMPs are usually under conditions of accurate control, and the failure to maintain this regulation is considered a component of arriving at a whole series of conditions. Examples of conditions in which MMPs are thought to be important include bone tissue repair, implantation of an embryo in the uterus, infiltration of immune cells into inflammatory areas, ovulation, spermatogenesis, tissue repair during wound healing, and organ differentiation, such as in venous and diabetic inflammation, pressure ulcers, colon ulcers, e.g. ulcerative colitis and Crohn's disease, duodenal ulcers, fibrosis, local tumor growth in neighboring areas, tumor cell metastases from primary or secondary areas, and tissue destruction in arthritis, skin diseases such as dystrophic epidermolysis bullosa, dermatitis herpetiformis, or conditions caused by complications of embolic phenomena, such as chronic or acute heart or brain infarctions.

Conditions in which MMP-3 and MMP-13 are involved include tissue destruction, such as in venous and diabetic wounds, pressure ulcers, colon ulcers, such as ulcerative colitis and Crohn's disease, and tissue destruction in arthritis, skin diseases, such as dystrophic epidermolysis bullosa, dermatitis herpetiformis, or conditions complicated by embolic phenomena, such as chronic or acute cardiac or cerebral infarctions.

Another important function of certain MMPs is to activate other enzymes, including other MMPs, by cleaving the prodomain from their protease domains. Thus, certain MMPs act in the regulation of the activity of other MMPs, in such a way that excessive production in one MMP can lead to excessive proteolysis of the extracellular matrix by another.

There is an opinion that the excessive production of MMP-3 is responsible for the pathological deterioration of tissues, which is subject to a number of diseases and conditions. For example, MMP-3 has been found in the synovium and cartilage of patients with osteoarthritis and rheumatoid arthritis, thereby implicating MMP-3 in joint damage in these diseases (see: K.L. Sirum, C.E. Brinkerhoff: "Biochemistry", 28, 8691, 1989; and Z. Gunja-Smith, H. Nagasse, J.F. Woessner: "Biochem. J.", 258, 115, 1989). MMP is also thought to play an important role in the pathology of osteoarthritis and rheumatoid arthritis (M. Stahle-Backdahle, B. Sandstedt, K. Bruce, A. Lindahl, M.G. Jimenez, J.A. Vega, C. Lopez Otin: "Lab.Invest." , 76, 717-28, 1997; and O. Lindy, Y.T. Konttinen, T. Sorsa, Y. Ding, S. Santavirta, A. Ceponis, C. Lopez Otin: "Arthritis Rheum.", 40, 1391-9 , 1997).

Overexpression of MMP-3 is also thought to be responsible for much of the tissue damage and chronicity of wounds, both venous and diabetic, as well as pressure ulcers (see M. Vaalamo, M. Weckroth, P. Puoakkainen, J. Kere, P. Saarinen, J. Lauharanta, U.K. Saarialho-Kere, "Brit. J. Dermotologya", 135, 52-59, 1996).

During normal and chronic wound healing, MMP-1 is expressed by migrating keratocytes at the wound edges (U.K. Saarialho-Kere, S.O. Kovacs, A.P. Pentland: "J. Clin. Invest.", 92, 2858-66, 1993). There is evidence to suggest that MMP-1 is required for keratocyte migration on type I collagen matrix in vitro, and that it is completely inhibited by the presence of the non-selective MMP inhibitor SC44463 ((N4-hydroxy)-N1-[(1S)-2-(4- methoxyphenyl)methyl-1-((1R)-methylamino)carbonyl)]-(2R)-2-(2-methylpropyl)butanediamide) (B.K. Pilcher, J.A. Dumin, B.D. Sudbeck, S.M. Krane, H.G. Welgus, W.C. Parks: " J. Cell Biol.", 137, 1-13, 1997). Keratocyte migration in vivo is essential for efficient wound healing.

MMP-2 and MMP-9 appear to play important roles in wound healing during the extended remodeling and initiation phases of re-epithelialization, respectively (M.S. Agren: "Brit. J. Dermatology", 131, 634-40, 1994; T. Salo, M. Mäkänen, M. Kylmäniemi: "Lab. Ivest.", 70, 176-82, 1994.). Potent, non-selective MMP inhibitor BB94 ((2S,3R)-5-methyl-3-{[(1S)-1-(methylcarbamoyl)-2-phenylethyl]carbamoyl}-2-[(2-thienylthio)methyl]hexanohydroxamic acid , batimastat), inhibits the invasion of endothelial cells of the diaphragm, thus inhibiting angiogenesis (G. Tarboletti, A. Garofalo, D. Belotti, T. Drudis, P. Borsotti, E. Scanziani, P.D. Brown, R. Giavazzi: "J. Natl. Cancer Inst.", 87, 293-8, 1995). There is evidence that this process requires active MMP-2 and/or 9.

Consequently, non-selective MMP inhibitors, which inhibit MMPs 1 and/or 2 and/or 9, can be expected to interfere with wound healing. As described above, MMP-14 is responsible for activating MMP-2, so MMP-14 can also interfere with wound healing.

MMP-3 production is also thought to be responsible for tissue damage in conditions where there is ulceration of the colon (such as ulcerative colitis and Crohn's disease, see: S.L. Pender, S.P. Tickle, A.J. Docherty, D. Howie, N.C. Wathen, T.T. MacDonald: "J. Immunol.", 158, 1582, 1997; C.J. Bailey, R.M. Hembry, A. Alexander, M.H. Irving, M.E. Grant, C.A. Shuttleworth: "J. Clin. Path.", 47, 113-6, 1994), or duodenal ulcer (see: U.K. Saarialho-Kere, M. Vaalamo, P. Puolakkainen, K. Airola, W.C. Parks, M.L. Karjalainen-Lindsberg: "Am. J. Pathol.", 148, 519-26, 1996). It is likely that MMP-1 and MMP-2 are required during the healing phase of these conditions. A selective MMP-3 inhibitor would be more effective than a non-selective one.

MMP-3 is also associated with skin diseases, such as dystrophic epidermolysis bullosa (T. Sato, K. Nomura, I. Hashimoto: "Arch. Dermatol. Res.", 287, 428, 1995) and dermatitis herpetiformis (K . Airola, M. Vaalamo, T. Reunala, U.K. Saarialho-Kere: "J. Invest. Dermatology", 105, 184-9, 1995.).

Rupture of atherosclerotic plaques by MMP-3 can lead to heart attack or stroke. (F. Mach et al.: "Circulation", 96, 396-9, 1997). Thus, MMP-3 inhibitors may find useful application in the treatment of conditions caused by or due to complications of embolic phenomena, such as chronic or acute heart or brain infarctions.

MMP-12 (macrophage elastase) is believed to contribute to the pathology of atherosclerosis, gastrointestinal ulcers, and emphysema. For example, in a rabbit model of atherosclerosis development, MMP-12 was expressed significantly by macrophage foam cells (S. Matsumoto et al.: "Am. J. Pathol.", 153, 109, 1998). MMP-12 is also highly expressed by macrophages surrounding the mucosal epithelium in human gastrointestinal ulcers, such as those found in patients with ulcerative colitis and Crohn's disease (M. Vaalamo et al.: "Am. J. Pathol. ", 152, 1005, 1998). MMP-12 is thought to be important in the development of lung damage in smokers. In a model of tobacco smoke-induced emphysema, mice lacking the MMP-12 gene were resistant to the condition, while wild-type mice experienced significant lung damage (R.D. Hautamaki et al.: "Science", 277, 2002, 1997).

For recent reports on MMPs see Zask et al.: "Current Pharmaceutical Design", 2, 624-661, 1996); Beckett: "Exp. Opin. Ther. Patents", 6, 1305-1315, 1996); and Beckett et al.: "Drug Discovery Today", Vol. 1 (No. 1), 16-26, 1996.

Alternate names of various MMPs and the substrates they act on are shown in the table below (Zask et al., as above).

[image]

Numerous publications, including those mentioned above, describe compounds of the general formula shown below as MMP inhibitors.

[image]

where "A" is known as an "alpha" group, and XCO is a group, which binds zinc, as half of a carboxylic or hydroxamic acid.

The review by Beckett et al., mentioned above, argues that a large range of groups can be tolerated at the P2' position, without significantly affecting compound behavior.

International patent applications no. WO96/33165 and WO96/33161 (British Biotech Pharmaceuticals Ltd.) disclose, in a general sense, compounds of the above formula "GENMMP", in which, inter alia, P1 is optionally substituted with phenyl(C1-C6)alkyl, and P3' is the group CHRxRy where Rx and Ry represent optionally substituted phenyl or heteroaryl rings. These compounds are believed to be selective inhibitors of MMP-3 and MMP-7 over human fibroblast collagenase (MMP-1) and 72KDa gelatinase (MMP-2).

International patent application WO96/16027 (Syntex Inc. and Agouron Phramaceuticals Inc.) discloses MMP inhibitors of the general formula "GENMMP", as shown above, where COX contains CO2H and CONHOH, P1' is the group R2X, which optionally contains a substituted aryl( C0-4 alkylene), and P3' is (CH2)pR7, where p is 0 to 4, provided that when p is 0, then R2X is biphenylalkyl and R7 is aryl or heteroaryl. Compounds where p is 0, 2 or 3 are preferred, and compounds where p is 0 and COX is CO2H or CONHOH are preferred for inhibiting matrilysin (ie, MMP-7).

A number of compounds from the aforementioned publication WO96/16027, general formula "GENMMP" were published by the Agouron group (2nd Winter Conference on Medicinal and Bioorganic Chemistry, Steamboat Springs, Colorado, USA, January 1997), e.g., where X is OH, P1' is an optionally 4'-substituted biarylpropyl group, P2' is isobutyl, and P3' is 4-methoxycarbonylphenyl. These compounds have been reported to possess weak to moderate selectivity for MMP-3/MMP-2. The use of an alpha-substituent, as well as the replacement of half of the 4-methoxycarbonylphenyl group with a 4-methylthionylphenyl group, was found to significantly increase the selectivity for MMP-3/MMP-2.

The review by Beckett (above) also mentions an Agouron compound (#31 review) of the general formula "GENMMP", where X is OH, P1' is biphenylpropyl, P2' is t-butyl, and P3' is 4-pyridyl . This compound has almost identical Ki values for MMP-3 and MMP-2, so it is not selective for MMP-3 over MMP-2.

International Patent Application No. WO95/12603 (Syntex) discloses compounds, said to be MMP-3 and MMP-7 inhibitors, of the formula "GENMPM" above, wherein P3' is a substituted phenyl moiety and P1' includes arylalkyl.

We have now found a group of compounds as MMP inhibitors, with good activity against MMP-3, MMP-12 and MMP-13, and good selectivity against MMP-3 over other MMPs, such as MMP-1, 2, 9 and 14. For this group of compounds, it was discovered that the selectivity with regard to MMP-3 significantly depends on a certain combination of substituents P1' and P3', the influence of which could not be predicted from the above-mentioned state of the art.

Thus, according to the present invention, a compound of formula (I) is provided

[image]

and its pharmaceutically acceptable salts, where:

R 1 is H, OH, C 1-4 alkyl, C 1-4 alkoxy or C 1-4 alkenyl;

R2 is C1-6 alkyl, optionally substituted with fluoro, indolyl, imidazolyl, SO2(C1-4 alkyl), C5-7 cycloalkyl,

or optionally protected by OH, SH, CONH2, CO2H, NH2 or NHC(=NH)NH2 group, C5-7 cycloalkyl, optionally substituted with C1-6 alkyl,

or benzyl, optionally substituted with optionally protected OH, C1-6 alkoxy, benzyloxy or benzylthio,

where, if desired, the protective groups for the mentioned OH, SH, CONH2, NH2, and NHC(=NH)NH2 groups are chosen from

C1-6 alkyl, benzyl, C1-6 alkanoyl;

and where optionally selected protecting groups for CO2H are selected from C1-6 alkyl or benzyl,

R 3 , R 5 and R 6 are independently selected from H and F;

R 4 is CH 3 , Cl or F;

X is HO or HONH;

Y is a direct bond or O;

Z is one group of formula (a)

[image]

where R10 is C1-4 alkyl, C1-4 alkoxymethyl, hydroxy(C2-4 alkyl), carboxy(C1-4 alkyl) or (amino or dimethylamino) C2-4 alkyl,

R11 is phenyl, naphthyl or pyridyl, optionally substituted with up to 3 substituents, independently selected from halo and methyl;

or (b)

[image] ;

R 14 is H, OH, CH 3 or halo;

Ar is a group of formula (c), (d) or (e)

[image]

in which they are

A is N or CR12,

B is N or CR13,

provided that both A and B are not N;

R 7 and R 9 are each independently H or F;

R 8 , R 12 and R 13 are each independently H, CN, C 1-6 alkyl, hydroxy(C 1-6 alkyl), hydroxy(C 1-6 )alkoxy, C 1-6 alkoxy(C 1-4 ) alkoxy, (amino or dimethylamino) C 1 -6 alkyl, CONH2, OH, halo, C1-6 alkoxy, (C1-6 alkoxy)methyl, piperazinylcarbonyl, piperidinyl, C(NH2)=NOH or C(=NH)NHOH, provided that of R8, R12 and R13 at least two H.

"Alkyl" groups, including moieties of "alkoxy" groups and "alkenyl" groups, may be straight or branched, where the carbon atoms permit.

"Halo" stands for F, Cl, Br or I.

The compounds according to this invention are MMP inhibitors, and they are particularly strong and selective MMP-3 inhibitors, with particularly good selectivity towards MMP-1, 2, 9 and/or 14. In addition, certain compounds according to the invention possess beneficial properties of inhibitory activity towards MMP- 12 and/or MMP-13.

Preferably, R 1 is H, OH, C 1-4 alkyl or C 1-4 alkoxy.

More preferably, R 15 is H, OH, n-propyl or ethoxy.

Most preferably, R1 is H.

Preferably, R 2 is C 1-6 alkyl, optionally substituted with indolyl, C 1-6 alkylthio, SO 2 (C 1-4 alkyl), C 5-7 cycloalkyl, OH or SH.

C5-7 cycloalkyl is optionally substituted with C1-6 alkyl,

or R 2 is benzyl.

More preferably, R2 is C1-6 alkyl, optionally substituted with OH, SO2(C1-4 alkyl) or C5-7 cycloalkyl,

cyclohexyl optionally substituted with C1-6 alkyl,

or R 2 is benzyl.

More preferably, R 2 is cyclohexylmethyl, isopropyl, 1-methylcyclohexyl, t-butyl, C(CH 3 ) 2 SO 2 CH 3 , benzyl or C(CH 3 ) 2 OH.

Furthermore, even more preferably, R 2 is isopropyl, t-butyl or benzyl.

Most preferably, R2 is t-butyl.

Preferably, Z is a group of formula (a)

[image]

in which R10 is C1-4 alkyl, C1-4 alkoxymethyl or hydroxy(C2-4 alkyl), and R11 is phenyl or pyridyl, optionally substituted with up to 3 substituents independently selected from halo and methyl,

or Z is

[image]

More preferably, Z is a group of formula (a)

[image]

wherein R 10 is C 1-4 alkyl, C 1-4 alkoxymethyl or hydroxy(C 2-4 alkyl), and R 11 is phenyl, pyridin-4-yl or pyridin-3-yl,

or Z is

[image] .

More preferably, Z is a group of formula (a)

[image]

wherein R10 is CH3, CH2OCH3 or CH2OH, and R11 is phenyl, pyridin-4-yl or pyridin-3-yl,

or Z is:

[image] .

Most preferably, Z is a group of formula

[image] .

Preferably R3 is H.

Preferably R4 is F when Y is O.

Preferably R 4 is Cl or CH, when Y is a direct bond.

Preferably R5 is H.

Preferably R6 is H.

Preferably, Ar is a group of formula (c)

[image]

where

A is CR12, and B is CR13,

R7 and R9 are each independently H or F,

R 8 and R 13 are each independently H, F, Cl, CN, CONH 2 , CH 2 or OCH 3 , and

R 12 is H, C 1-6 alkyl, CN, hydroxy(C 2-6 alkyl), (amino or dimethylamino)C 2-6 alkyl, CONH 2 , OH, halo, C 1-6 alkoxy, (C 1-6 alkoxy)methyl, piperazinylcarbonyl, piperidinyl, C(NH2)NOH or C(=NH)NHOH.

More preferably, Ar is a group of formula (c)

[image]

where

A is CR12, B is CR13, R7, R8 and R9 are H;

R 12 is H, C 1-6 alkyl, CN, hydroxy(C 2-6 alkyl), (amino or dimethylamino)C 2-6 alkyl, CONH 2 , OH, halo, C 1-6 alkoxy, (C 1-6 alkoxy)methyl, C( NH 2 )=NOH or C(=NH)NHOH, and R 13 is H, OCH 3 , CN, CONH 2 , CH 3 or F.

Furthermore, more preferably, Ar is phenyl, 3-methoxyphenyl, 4-cyanophenyl, 3-cyanophenyl, 3-carbamoylphenyl or 4-hydroxyamidinophenyl.

Most preferably, Ar is phenyl or 3-methoxyphenyl.

A preferred group of substances is one in which the ingredients have the meanings mentioned in the Examples, i.e.:

R1 is H, OH, n-propyl or ethoxy,

R2 is t-butyl, isopropyl or benzyl,

Z is a formula group

[image]

wherein R 10 is CH 3 , CH 2 OCH 3 , or CH 2 OH, and R 11 is phenyl, pyridin-4-yl or pyridin-3-yl;

or Z is

[image]

where R 3 is H,

R4 is CH3, Cl or F,

R5 is H,

R 6 is H,

Ar is phenyl, 3-methoxyphenyl, 4-cyanophenyl, 3-cyanophenyl, 3-carbamoylphenyl or 4-hydroxyamidinophenyl, and salts thereof.

Another preferred group are the compounds described in the Examples, and their salts. The most preferred substances are selected from Examples 3, 4, 8, 14, 15, 16, 22, 29, 30, 31 and 32, as well as their salts.

Pharmaceutically acceptable salts are well known to those skilled in the art, and include, for example, those mentioned in the prior art above, and in Borge et al.: "J. Pharm. Sci.", 66, 1-19, 1997. Suitable addition acid salts are formed from acids , which form non-toxic salts and include salts of hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate, hydrogen phosphate, acetate, trifluoroacetate, gluconate, lactate, salicylate, citrate, tratrate, ascorbate, succinate, maleate, fumarate, gluconate, formate , benzoate,, methanesulfonate, ethanesulfonate, benzylsulfonate and p-toluenesulfonate.

Pharmaceutically acceptable base addition salts are well known to those skilled in the art, and include, for example, those in the prior art, and may be formed from bases that form non-toxic salts, and include aluminum, calcium, lithium, magnesium, potassium, sodium and zinc salts, and salts of non-toxic amines, such as diethylamine.

Certain compounds of formula (I) may exist as geometric isomers. Compounds of formula (I) may contain one or more asymmetric centers, in addition to those specified by formula (I), and thus exist in two or more stereoisometric forms. This invention includes all individual stereoisomers and geometric isomers of compounds of formula (I), in addition to certain centers in formula (I), including group Z, and mixtures thereof.

Another aspect of the invention is a pharmaceutical preparation with a compound or salt as defined above and a pharmaceutically acceptable vehicle, diluent or carrier.

Another aspect of the present invention is a compound or salt as defined above, which can be used as a medicament.

A further aspect of the present invention is the use of a compound or salt according to the above definitions for the manufacture of a medicament for the treatment of conditions created by one or more matrix metalloproteases, particularly MMP-3 and/or MMP-12 and/or MMP-13.

Another aspect of this invention is a method for treating a condition that is under the influence of one or more matrix metalloproteases, particularly MMP-3 and/or MMP-12 and/or MMP-13.

It should be accepted that the call for treatment includes prophylaxis as well as alleviation of established symptoms of conditions created by MMP.

In addition, the invention provides processes for the production of compounds according to the invention, which are described below and in the Examples. A person skilled in the art will know that the compounds according to the invention can be produced by other procedures, different from those specifically described here, by adapting the procedures given here in the following paragraphs and/or by adapting them, and procedures known in the art. Suitable guidelines for synthesis, transformation of functional groups, use of protecting groups, etc., are found, e.g., in R.C. Larock: "Comprehensive Organic Transformations", VCH Publishers Inc., 1989; J. March: "Advanced Organic Chemistry", Wiley Interscience, 1985; S. Warren: "Designing Organic Synthesis", Wiley Interscience, 1978; S. Warren: "Organic Synthesis. The Disconnection Approach", Wiley Interscience, 1982; R.K. Mackie and D.M. Smith: "Guidebook to Organic Synthesis", Longman, 1982; T.W. Greene and P.G.M. Wuts: "Protective Groups in Organic Synthesis", John Wiley and Sons Inc., 1991; and P.J. Kocienski: "Protecting Groups", Georg Thieme Verlag. in 1994

In the procedures described below, unless otherwise stated, the substituents are defined as above, with reference to the compound of formula (I).

Procedure 1

Compounds of formula (I), in which X is OH, can be obtained via the corresponding compound of formula (II), where X1 is a group capable of transformation into a carboxy group, under conditions that do not result from basic transformations of other parts of compound (II). A suitable example of such a group is CO2 (t-butyl or methyl). The t-butyl group can be cleaved off by reaction with an acid such as hydrochloric acid or trifluoroacetic acid (TFA), in a suitable solvent such as anhydrous dichloromethane or dioxane, and at a suitable temperature, eg between 0 and 20°C. The methyl ester can be hydrolyzed with a hydroxide such as lithium hydroxide, in a suitable solvent such as tetrahydrofuran/water, preferably at room temperature.

[image]

Compounds of formula (II) can be produced by methods known in the art, and as given below in the Examples, eg by coupling a suitable amine with acid derivatives, which can be obtained by known chemical methods.

Procedure 2

Compounds of formula (I) wherein X is NHOH may be prepared from the corresponding compounds of formula (I) wherein X is OH with hydroxylamine, such as by formation from a hydroxylamine salt such as the hydrochloride, with a suitable base such as a tertiary amine, eg diisopropylethylamine, in a suitable solvent such as N,N-dimethylformamide (DMF), using a coupling agent such as N-[(dimethylamino)-1H-1,2,3-triazolo[4,5-b]pyridine -1-ylmethylene]-N-methylmethanenium hexafluorophosphate N-oxide ("HATU", which is described in "Tet. Letts.", 35, 2279, 1994), at a suitable temperature of 0 to 20 °C.

Compounds of formula (I) in which X is OH can be prepared by conventional procedures and those described herein.

Procedure 3

Compounds of formula (I) in which X is NHOH can be obtained from the corresponding compounds of formula (I) in which X is OH by treating the compound of formula (I) with O-allylhydroxylamine, such as by forming from an allylhydroxylamine salt such as the hydrochloride, with with a suitable base such as a tertiary amine, eg diisopropylethylamine, in a suitable solvent such as N,N-dimethylformamide (DMF) or dichloromethane, and with a coupling agent, eg a peptide coupling agent, such as 7-azabenzotriazol-1-yloxytris (pyrrolidino)phosphonium hexafluorophosphate ("PyAOP"), at a suitable temperature of 0 to 20 °C. This gives compounds of formula (III) below.

This bonding procedure is described in general terms in "Tet. Letts.", 35, 2279, 1994.

[image]

Compounds of formula (III) can be transformed into compounds of formula (I) wherein X is NHOH, by treatment with ammonium formate in the presence of a suitable catalyst such as bis(triphenylphosphine) palladium(II) acetate, in a suitable solvent such as water-diluted ethanol, at a suitable temperature such as the reflux temperature of dilute ethanol.

Procedure 4

Compounds of formula (I) where X is NHOH and R 1 is OH can be prepared by reacting a compound of formula (IV) with a hydroxylamine, eg by forming a salt of the hydroxylamine such as the hydrochloride, with a suitable base such as sodium methoxide, in a suitable solvent such as which is methanol.

[image]

Compounds of formula (IV) can be prepared by conventional procedures, as given below in the Examples.

Procedure 5

Compounds of formula (I) can be produced from compounds of formula (V) by cross-coupling with a compound of formula (VI) below:

[image]

where X2 is a protected acid, such as a t-butyl or methyl ester group, and LG is a cross-linked group, which is isolated, such as I, Br or OSO2CF3. The cross-linking reaction can be carried out in the presence of a catalyst such as bis(tri-o-tolyl)phosphine palladium (II) acetate, with a suitable base such as triethylamine, in a suitable solvent such as acetonitrile or DMF, at a suitable temperature between 50 and 150 °C.

This type of reaction is described in general terms in Heck: "Tet. Letts.", 25, 2271, (1984), and in numerous other articles.

Compounds of formula (V) can be prepared by conventional procedures, as well as by adaptations of the procedures described in Preparations, below.

Compounds of formula (VI) can be prepared by conventional procedures, as well as by adaptations of the procedures described in Preparations below, and with reference to the following articles: "Synthesis", 709, (1984), "J. Chem. Soc. Perkin Trans I", 1841, (1977); "J. Org. Chem.", 59, 6095, (1994); ibid., 44, 4444, (1979), and Tet. Letts., 38, 1749, (1997).

The resulting product of this reaction is a mixture of compounds of formula (VIIa) and (VIIb), wherein X 2 is as defined above for compounds of formula (V).

[image]

Compounds of formulas (VIIa) and (VIIb) can be transformed into a compound of formula (I), where X is OH, by reduction of the olefinic bond, using conventional procedures, such as hydrogenation in the presence of a catalyst, or by reaction with a diimide, which can be formed, eg from p-toluenesulfonyl hydrazide, and by deprotection from the protected acidic half of X2.

It will be clear to those skilled in the art that other regimes of protection and deprotection thereafter, during the synthesis of the compounds of the invention can be achieved by conventional techniques, eg as described in the books by Green and Wuts, and Kocienski, cited above.

When desired or necessary, the compound of formula (I) is converted into its pharmaceutically acceptable salt. A pharmaceutically acceptable salt of a compound of formula (I) may be readily prepared by mixing a solution of compound (I) with the desired acid or base, as appropriate. The salt can be precipitated out of solution and collected by filtration, or it can be collected by some other means, such as evaporation of the solvent.

Some compounds of the invention can be converted into certain other compounds of the invention by procedures well known in the literature.

The compounds of the invention may be prepared using any of the procedures described herein in the Procedures and Examples, or by suitable adaptations thereof, using those skilled in the art. It should be understood that the synthetic transformation procedures mentioned here can be carried out in a different order, with the aim of efficiently synthesizing the desired compounds. A skilled chemist will try his or her experience as to the most efficient order of reactions to synthesize the target compound.

The compounds and salts of this invention can be isolated and purified by conventional methods.

Separation of the diastereoisomers can be achieved by conventional techniques, eg fractional crystallization, or by HPLC (high pressure liquid chromatography) of a stereoisomeric mixture of a compound of formula (I), or a suitable salt or derivative. A single enantiomer of a compound of formula (I) can also be prepared from a suitable optically pure intermediate either by resolution, eg by HPLC of the corresponding racemate, using a suitable chiral support, or by fractional crystallization of a diastereoisomeric salt formed by reaction of the corresponding racemate with a suitable optically active acid or base.

For human use, the compounds of formula (I) or their salts can be administered alone, but in general they will be administered as a mixture with a pharmaceutically acceptable diluent or carrier, from the point of view of the planned route of administration and standard pharmaceutical practice. For example, they can be administered orally, including sublingually, in the form of tablets containing additives, such as starch or lactose, or in capsules or ovules, alone or mixed with additives, or in the form of syrups, solutions or suspensions, containing fragrance and coloring agents. The compound or salt can be inserted into capsules or tablets with targeted administration in the intestinal tract or duodenum, with delayed dissolution of said capsules or tablets for a certain time after oral administration. Dissolution can be controlled by the sensitivity of the bacteria found in the duodenum or tract, so that the main dissolution does not occur before the medicinal substance reaches the target site of the tract.

The compounds or salts may be administered by injection, eg, intravenously, intramuscularly or subcutaneously. For parenteral administration, they are best administered in the form of sterile aqueous solutions or suspensions, which may contain other substances, eg, enough salt or glucose to make the solution isotonic with blood. They can be administered locally, in the form of sterile creams, suspensions, lotions, ointments, powders, sprays, by inserting into medicine, or through skin patches. For example, they can be included in a cream, consisting of an aqueous or oily emulsion of polyethylene glycol or liquid paraffin, or they can be included in a spread, consisting of a base of white paraffin wax, or as a hydrogel with cellulose or polyacrylate derivatives or other viscosity modifiers, or as a dry powder or liquid spray or aerosol with butane/propane, HFA or CFC propellants, or as inclusion in drugs, either as mesh coatings (screens), white soft paraffin or polyethylene glycol coated gauze screens, or hydrogel , hydrocolloid, alginate or as thin (film) coatings. The compound or salt could also be administered to the eyes, in the form of eye drops with suitable buffers, viscosity modifiers (e.g. cellulose derivatives), preservatives (e.g. benzalkonium chloride (BZK)) and strength adjusting agents (e.g. sodium chloride). . Such formulation techniques are well known in the art.

All these formulations may also contain appropriate stabilizers and preservatives.

For oral and parenteral administration to human patients, daily dosage levels of the compound of formula (I) or salts thereof are from 0.001 to 20, preferably from 0.01 to 20, more preferably from 0.1 to 10, and most preferably from 0.5 to 5 mg/kg (in single or divided doses). Thus, the tablets or capsules will contain from 0.1 to 500, preferably from 50 to 200, mg of the active compound for administration at once or in two or more doses each time, as needed.

For local administration to human patients with chronic wounds, the daily dose of the compounds in suspensions or other formulations may be from 0.01 to 50 mg/ml, preferably from 0.3 to 30 mg/ml.

In each individual case, the doctor will determine the actual dose, which will be the most suitable for the individual patient, and will depend on the age, weight and response of each patient. The above doses are examples for intermediate cases. It is understood that there may be individual cases, where higher or lower doses are prescribed, and they are also within the scope of this invention.

TEST PROCEDURES

The ability of the compounds to prevent cleavage of fluorogenic peptides by MMPs 1, 2, 3, 9, 12, 13 and 14 are described below.

Assays for MMP 2, 3, 9, and 14 are based on the original protocol described by Knight et al. ("Fed. Euro. Biochem. Soc.", 296(3), 263-266, (1992)), with minor modifications given below.

Inhibition of MMP-1

(i) Preparation of enzymes

The catalytic domain of MMP-1 was prepared in Pfizer's main research laboratory. The MMP-1 matrix solution (1 µM) was activated by the addition of aminophenol mercury acetate (APMA), with a final concentration of 1 mM, for 20 minutes at 37 °C. MMP-1 was then diluted in Tris-HCl assay buffer (50 mM Tris, 200 mM NaCl, 5 mM CaCl2, 20 µM ZnSO4, 0.05% Brij 35) pH 7.5, down to a concentration of 10 nM. The final enzyme concentration in the assay was 1 nM.

(ii) Substrate

The fluorogenic substrate used in this assay was Dnp-Pro-�-cyclohexyl-Ala-Gly-Cys(Me)-His-Ala-Lys(N-Me-Ala)-NH2, as originally described by Bickett et al ("Anal . Biochem.", 212, 58-64, (1993)). The final substrate concentration used in this assay was 10 µM.

(iii) Determination of enzyme inhibition

Test compounds were dissolved in dimethyl sulfoxide and diluted with assay buffer so that no more than 1% dimethyl sulfoxide was present. Test compound and enzyme were loaded into each well of a 96-well plate, and allowed to equilibrate for 15 minutes at 37 °C on a rotary shaker, before addition of substrate. The plates were then incubated for 1 hour at 37 °C before determination of fluorescence (substrate extraction) using a fluorimeter (Fluostar; BMG Lab Technologies, Aylesbury, UK), at an excitation wavelength of 355 nm and an emission wavelength of 440 nm. The strength of the inhibitor was measured through the amount of cleaved substrate, obtained when using a certain range of compound concentrations, and the IC50 value (concentration necessary to inhibit 50% of enzyme activity) was calculated from the obtained dose-response curve.

Inhibition of MMP-2, MMP-3 and MMP-9

(i) Preparation of enzymes

The catalytic domains of MMP-2, MMP-3, and MMP-9 were prepared in Pfizer's main research laboratory. The matrix solution of MMP-2, MMP-3 or MMP-9 (1 µM) was activated by the addition of aminophenyl mercury acetate (APMA). For MMP-2 and MMP-9, a final concentration of 1 mM APMA was added, followed by incubation for 1 hour at 37 °C. MMP-3 was activated by the addition of 2 mM APMA and then incubated for 3 hours at 37 °C. The enzymes were then diluted with Tris-HCl assay buffer (100 mM Tris, 100 mM NaCl, 10 mM CaCl and 0.16% Brij 35, pH 7.5), to a concentration of 10 nM. The final concentration of the enzyme used in the tests was 1 nM.

(ii) Substrate

The fluorogenic substrate used in this assay was Mca-Arg-Pro-Lys-Pro-Tyr-Ala-Nva-Trp-Met-Lys(Dnp)-NH2 (Bachem Ltd, Essex, UK), as originally described by Nagase et al. ("J. Biol. Chem.", 269 (33), 20952-20957, (1994)). This substrate was chosen because it has a balanced rate of hydrolysis compared to MMP 2, 3 and 9 (kcat/km 54,000, 59,400 and 55,300 s-1 M-1, respectively). The final substrate concentration in this assay was 5 µM.

(iii) Determination of enzyme inhibition

Test compounds were dissolved in dimethyl sulfoxide and diluted with test buffer solution (as above) so that no more than 1% dimethyl sulfoxide remained. Test compound and enzyme were loaded into each well of a 96-well plate, and allowed to equilibrate for 15 min at 37 °C on a rotary shaker before addition of substrate. The plates were then incubated for 1 hour at 37 °C before fluorescence determination with a fluorimeter (Fluostar, BMG Lab Technologies, Aylesbury, UK), at an excitation wavelength of 328 nm and an emission wavelength of 393 nm. The potency of inhibition was measured by the amount of substrate cleaved, using a range of compound concentrations, and the IC50 value was calculated.

Inhibition of MMP-12 (Elastase of human macrophages)

(i) Preparation of enzymes

The catalytic domain of MMP-12 (200 µg/ml) was used. MMP-12 was diluted in Tris-HCl assay buffer (50 mM Tris, 200 mM NaCl, 5 mM CaCl2, 20 µM ZnSO4, 0.02% Brij 35) pH 7.4 to 240 ng/ml. The final concentration of the enzyme used was 60 ng/ml.

(ii) Substrate

The fluorogenic substrate used in this assay was DNP-Pro-Cha-Gly-Cys(Me)-His-Ala-Lys(NMA)-NH2. The final substrate concentration in this assay was 10 µM.

(iii) Determination of enzyme inhibition

Test compounds were dissolved in dimethyl sulfoxide and diluted with assay buffer to a dimethyl sulfoxide residue of less than 1%. Test compound and enzyme were loaded into each well of a 96-well plate, and allowed to equilibrate for 15 minutes at room temperature on a rotary shaker before addition of substrate. The plates were then incubated for 2 hours at room temperature before determining fluorescence (substrate cleavage) using a fluorimeter at an excitation wavelength of 360 nm and an emission wavelength of 460 nm. The potency of the inhibitor was measured from the amount of cleaved substrate using various concentration ranges of the tested compounds, and the IC50 value was calculated from the obtained dose-response curve.

Inhibition of MMP-13

(i) Preparation of enzymes

Human recombinant MMP-13 was produced by PanVera Corporation (Madison, Wisconsin) and characterized by Pfizer (Groton, CT). 1.9 mg/ml matrix solution was activated with 2 mM APMA for 2 hours at 37 °C. MMP-13 was then diluted with assay buffer (50 mM Tris, 200 mM NaCl, 5 mM CaCl2, 20 mM ZnCl2, and 0.02% Brij 35) at pH 7.5 to a concentration of 5.3 nM. The final enzyme concentration in the test was 1.3 nM.

(ii) Substrate

The fluorogenic substrate used in this assay was Dup-Pro-Cha-Gly-Cys(Me)-His-Ala-Lys(NMA)-NH2. The final substrate concentration was 10 µM in this test.

(iii) Determination of enzyme inhibition

Test compounds were dissolved in dimethyl sulfoxide and diluted with assay buffer so that less than 1% dimethyl sulfoxide remained. Test compound and enzyme were loaded into each well of a 96-well plate. Adding the substrate to each well started the reaction. Fluorescence intensity was determined in a 96-well plate with a fluorometer (Cytofluor II; PerSeptive Biosystems Inc., Framingham, MA), at an excitation wavelength of 360 nm and an emission wavelength of 460 nm. The strength of the inhibitor was measured using the amount of cleaved substrate, obtained from a certain range of concentrations of the tested compounds, and the IC50 value was calculated from the obtained dose-response curve.

Inhibition of MMP-14

(i) Preparation of enzymes

The catalytic domain of MMP-14 was purchased from prof. Tschesche, Department of Biochemistry, University of Bielefeld, Germany. 10 µM enzyme matrix solution was activated for 20 minutes at 25 °C, after addition of 5 µg/ml trypsin (Sigma, Dorset, Great Britain). Trypsin activity was then neutralized by the addition of 50 µg/ml soybean trypsin (Sigma, Dorset, UK), before diluting this enzyme solution in Tris-HCl buffer (100 mM Tris, 100 mM NaCl, 10 mM CaCl2 and 0.16% Brij 35, pH 7.5) to a concentration of 10 nM. The final concentration in this assay was 1 nM.

(ii) Substrate

The fluorogenic substrate used in this assay was Mca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 (Bachem Ltd, Essex, UK), as described by Will et al. ("J. Biol. Chem." , 271 (29), 17119-17123, (1996)). The final substrate concentration in this assay was 10 µM.

Determination of enzyme inhibition of the test compounds was performed as described for MMP-2, 3 and 9.

Some data on the activity of certain compounds from the Examples are given in the table below.

"MMP-x" inhibitor IC50/nM

[image]

*Selectivity of MMP-3 over MMP-2

The compounds of Examples 6, 8, 9, 14, 15 and 22 had IC50 values for MMP-3 in the range of 8 to 65 nM, and MMP-3/MMP-2 selectivities in the range of 195 to 930.

EXAMPLES AND PREPARATIONS

Melting points (M.p.) were determined using open glass capillary tubes and a Gallenkamp M.p. apparatus, and the values are uncorrected. Nuclear magnetic resonance (NMR) spectra were obtained using Varian Unity Inova-400, Varian Unity Inova-300, or Brucker AC300 spectrometers, and are given in parts per million (ppm) relative to tetramethylsilane. Mass spectrum (MS) data were obtained on a Finnigan Mat. TSQ 7000 or on Fisons Instruments Trio 1000. Calculated and observed ions refer to the lowest mass isotopic compositions. Infrared (IR) spectra were measured using a Nicolet Magna 550 Fourier transform infrared spectrometer. Flash chromatography refers to column chromatography over silica gel (Kieselgel 60, grain size 230-400, from E. Merck, Darmstadt, Germany). Kieselgel 60 F254 plates from E. Merck were used for TLC, and compounds were visualized by ultraviolet (UV) light, with 5% potassium permanganate or Dragendorff's reagent (sprayed with aqueous sodium nitrite). Hexane refers to a mixture of hexanes (HPLC grade), boiling point 65-70 °C. Ether means diethyl ether. Acetic acid means glacial acetic acid. 1-hydroxy-7-aza-1H-1,2,3-benzotriazole (HOAt), N-[(diethylamino)-1H-1,2,3-triazolo[4,5]pyridin-1-ylmethylene]-N -methylmethaninium hexafluorophosphate-N-oxide (HATU) and 7-azabenzotriazol-1-yloxytris(pyrrolidino)phosphonium hexafluorophosphate (PyAOP) were purchased from PerSeptive Biosystems, Great Britain, Ltd. "DIPE" means diisopropyl ether. Silica gel for reverse phase flash chromatography was purchased from Fluka (Fluka 100, C18, 40-63 µ). Pentane means the HPLC grade of n-pentane (blurring point 35-37 °C).

ATTENTION: Some 4-aminobiphenyl derivatives are described in Preparations below. 4-aminobiphenyl is a known human carcinogen. That is why its analogues should be treated with due care. For the most important references, see K. Yuta, P.C. Jurs: "J. Med. Chem." 24(3), 241-51, (1981); Z. You, M.D. Brezzell, S.K. Das, B.H. Hooberman, J.E. Sinsheimer: "Mutat. Res.", 320(1-2), 45-58, (1994); and S.S. Hecht et al.: "J. Med. Chem.", 22(8), 981-7, (1979)

Example 1

(3R)-3-({[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]amino}carbonyl)-6-[(3-methyl -4-phenyl)phenyl]hexanoic acid

[image]

Trifluoroacetic acid (5 ml) was added dropwise over 5 minutes to a stirred solution of tert-butyl (3R)-3-({[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl) ]amino}carbonyl)propyl]amino}carbonyl)-6-[3-methyl-(4-phenyl)phenyl] hexanoate (preparation 3) (285 mg, 0.47 mmol) in anhydrous dichloromethane (5 mL) under nitrogen at 20 °C. The mixture was stirred for 4 hours and concentrated under reduced pressure. The residue was dissolved in toluene and concentrated under reduced pressure (2×), then triturated with ether to give a colorless solid (210 mg, 82 %). T.t. 160-162 °C (from ethyl acetate).

Rf 0.17 (hexane/ether (acetic acid = 50:50:1).

Retention time in HPLC 7.3 minutes (Phenomenex Magellen C18 silanized silica gel (5 µ), eluting with acetonitrile/water/trifluoroacetic acid = 70:30:0.1 (1 ml/minute), detection with UV light (220 nm).

δH (400 MHz, CD3OD) (substitutable hydrogens only partially substituted) 1.02 (9H, s); 1.44 (3H, d, J=7 Hz); 1.52 (4H, m); 2.16 (3H, s); 2.37 (1H, dd, J=4 and 15 Hz); 2.50 (2H, m); 2.60 (1H, dd, J=9 and 15 Hz), 2.84 (1H, m); 4.37 (1H, d, J=10 Hz); 4.99 (1H, pentet, J=7 Hz); 6.94 (1H, d, J=8 Hz); 7.00 (2H, s and d, J=8 Hz, overlapping); 7.13 (1H, t, J=7 Hz); 7.16 (2H, t, J=7 Hz), 7.23 (4H, t, J=7 Hz); 7.29 (1H, t, J=7 Hz); 7.38 (2H, t, J=7 Hz); 7.72 (1H, no d); 8.45 (1H, number d).

LRMS (thermospray) m/z = 543 (MH+)

FTIR νmax (KBr disc) 3290; 2980; 2930; 1707; 1661; 1633; 1553; 700 cm-1.

Found: C, 75.22; H, 7.73; N, 5,16.

C34H42N2O4 required: C, 75.25; H, 7.80; N, 5.16%.

Example 2

(2R)-2-{3-[3-chloro-(4-phenyl)phenyl]propyl}-N1-{(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl] amino}carbonyl)propyl]-(N4-hydroxy)butane diamide

[image]

a) O-allylhydroxylamine hydrochloride (17 mg, 0.152 mmol) was added to a stirred solution of (3R)-6-[(3-chloro-4-phenyl)phenyl]-3-({[(1S)-2,2-dimethyl -1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]amino}carbonyl)hexanoic acid (preparation 4) (66 mg, 0.117 mmol) and diisopropylethylamine (78 µl, 0.456 mmol) in anhydrous dichloromethane ( 1 ml) under nitrogen at 0 °C. 7-Azabenzotriazol-1-yloxytris(pyrrolidino)phosphonium hexafluorophosphate (79 mg, 0.152 mmol) was added all at once, and the mixture was stirred for 2 hours at 0 °C and then allowed to warm to room temperature. After an additional 1 hour, the mixture was poured into ethyl acetate (25 ml) and washed, respectively, with 5% aqueous citric acid (2 x 10 ml) and saturated aqueous sodium bicarbonate (2 x 10 ml). The organic solution was dried (Na2SO4) and concentrated under reduced pressure. The solid residue was suspended in ether (3 mL) and filtered to give (2R)-2-{3-[3-chloro-(4-phenyl)phenyl]propyl}-N1-[(1S)-2,2 -dimethyl-1-({[(1R)-1-phenylethyl]amino}amino}carbonyl)propyl]-(N4-3-propenyloxy)butanediamide (58 mg, 82%) as a white solid.

Rf 0.45 (ethyl acetate:hexane = 2:1).

δH (400 MHz, CDCl 3 ) 0.98 (9H, s); 1.46 (3H, d, J=6.5 Hz); 1.48 (4H, m); 2.17 (1H, m); 2.39 (1H, m); 2.50 (2H, m); 2.74 (1H, m); 4.13 (1H, d, J=8 Hz); 4.29 (2H, d, J=6 Hz); 5.06 (1H, pentet, J=6.5 Hz); 5.29 (2H, no d); 5.87 (2H, m and br d, overlap); 6.45 (1H, no s); 6.97 (1H, d, J=8 Hz); 7.19 (7H, complex); 7.35 (5H, complex); 8.36 (1H, no s).

LRMS (thermospray) m/z = 618 (MH+, weak).

b) Mixed mixture (2R)-2-{3-[3-chloro(4-phenyl)phenyl]propyl}-N1-[(1S)-2,2-dimethyl-1-({[(1R)-1 -phenylethyl]amino}carbonyl)propyl]-(N4-3-propenyloxy)butanediamide (56 mg, 0.091 mmol) and ammonium formate (59 mg, 0.93 mmol) in ethanol/water (4:1, 4 mL) heated was refluxed under nitrogen and a colorless solution was obtained. Bis(triphenylphosphine)palladium acetate (3.4 mg, 0.00465 mmol) was added, and the mixture was heated at reflux for 40 minutes. After cooling, the brown solution was diluted with ethyl acetate (25 ml), washed with saturated aqueous sodium chloride solution (2 x 10 ml), dried (Na2SO4) and concentrated under reduced pressure. The residue was purified by column chromatography (C18 silanized silica gel 40-63 µ, eluting with methanol:water = 5:1) and obtained (2R)-2-{3-[3-chloro-(4-phenyl)phenyl]propyl }-N1-[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]-(N4-hydroxy)butanediamide (46 mg, 88%) as colorless solids.

T.t. 107-109 °C.

Rf 0.43 (C18 silanized silica gel, methanol:water = 5:1)

δH (400 MHz, CD3OD) 0.98 (9H,s); 1.39 (3H, d, J=6.5 Hz); 1.47 (4H, m); 2.13 (1H, dd, J=6 and 14 Hz); 2.31 (1H, dd, J=9 and 14 Hz); 2.48 (2H, m); 2.85 (1H, m); 4.32 (1H, s); 4.96 (1H, q, J=6.5 Hz); 6.80 (1H, d, J=8 Hz); 7.11 (4H, complex); 7.22 (3H, complex); 7.32 (5H, complex).

LRMS (thermospray) m/z = 578 (MH+, weak); 562 (MH+ - O).

Found: C, 67.69; H, 6.90; N, 7,19.

C33H40ClN3O4 • 0.5 H2O requirements: C, 67.51; H, 7.04; N, 7.16%.

Example 3

N1-[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]-(N4)hydroxy)-(2R)-2-{3-[3 -methyl-(4-phenyl)phenyl]propyl}butanediamide

[image]

a) O-allylhydroxylamine hydrochloride (295 mg, 2.69 mmol) was added to a stirred solution of (3R)-3-({[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl) ]amino}carbonyl)propyl]amino}carbonyl)-6-[(3-methyl-4-phenyl)phenyl]-hexanoic acid (Example 1) (974 mg, 1.79 mmol) and diisopropylethylamine (1.56 ml, 8.97 mmol) in anhydrous dimethylformamide (25 ml) under nitrogen at 0 °C. Then, 7-azabenzotriazol-1-yloxytris(pyrrolidino)phosphonium hexafluorophosphate (1.40 g, 2.69 mmol) was added all at once and the mixture was allowed to warm to room temperature. After another 4.25 hours, the mixture was poured into ethyl acetate (500 mL) and washed with saturated aqueous sodium bicarbonate (2 x 200 mL). The washed parts were extracted with ethyl acetate (2 × 100 ml). The combined organic solvents were dried (Na2SO4) and concentrated under reduced pressure. The solid residue was suspended in ether (3 mL) and filtered, and then recrystallized from ethyl acetate to give N1-[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino} carbonyl)propyl]-(2R)-2-{3-[3-methyl-(4-phenyl)phenyl]propyl}-(N4-3-propenyloxy)butanediamide (640 mg, 60%) as a white solid.

Rf 0.31 (ethyl acetate/hexane = 1:1).

δH (400 MHz, CDCl 3 ) 1.00 (9H, s); 1.48 (3H, d, J=6.5 Hz); 1.56 (4H, m); 2.22 (3H, s); 2.42 (1H, m); 2.52 (3H, m); 2.80 (1H, m); 4.20 (1H, d, J=8 Hz); 4.29 (2H, d, J=5 Hz); 5.07 (1H, pentet, J=6.5 Hz); 5.28 (2H, no); 5.90 (1H, m); 6.13 (1H, no s); 6.61 (1H, no d); 6.95 (1H, d, J=8 Hz); 7.00 (1H, d, J=8 Hz); 7.25 (8H, complex); 7.38 (2H, t, J=7 Hz); 8.69 (1H, no s).

LRMS (thermospray) m/z = 598 (MH+, weak); 542 (MH2+ - allyloxy, peak base).

b) Mixed mixture N1-[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]-(2R)-2-{3-[3-methyl -(4-phenyl)phenyl]propyl}-(N4-3-propenyloxy)butanediamide (794 mg, 1.32 mmol) and ammonium formate (419 mg, 6.64 mmol) in ethanol/water (4:1, 25 ml) was heated at reflux under nitrogen, and a colorless solution was obtained. Bis(triphenylphosphine)palladium acetate (40 mg, 0.066 mmol) was added, and the mixture was heated at reflux for 90 min. After cooling, the brown solution was diluted with ethyl acetate (250 ml) and washed with saturated aqueous sodium chloride solution (2 x 100 ml), dried (Na 2 SO 4 ) and concentrated under reduced pressure. The residue was purified by column chromatography (C18 silanized silica gel, eluting with methanol:water = 5:1), and then by trituration with methanol/diisopropyl ether, N1-[(1S)-2,2-dimethyl-1-({[(1R )-1-phenylethyl]amino}carbonyl)propyl]-(N4-hydroxy)-(2R)-2-{3-[3-methyl-(4-phenyl)phenyl]propyl}butanediamide (440 mg, 59 %) in the form of a colorless solid.

T.t. 114-116.5 °C.

Rf 0.23 (C18 silanized silica gel, methanol:water=5:1).

δH (400MHz, DMSO-d6) 0.85 (9H, s); 1.24 (3H, d, J=7 Hz); 1.31 (1H, m); 1.44 (3H, m); 1.98 (1H, dd, J=7 and 13 Hz); 2.11 (1H, m, and 3H, s, overlap); 2.42 (2H, m); 2.77 (1H, m); 4.26 (1H, d, 10 Hz); 4.87 (1H, pentet, J=7 Hz); 6.91 (1H, d, J=8 Hz); 6.97 (2H, s and d, J=8Hz, overlapping); 7.18 (7H, complex); 7.29 (1H, t, J=6 Hz); 7.35 (2H, t, J=8 Hz); 7.64 (1H, no d); 8.23 (1H, no d); 8.44 (1H, s); 10.28 (1H, no s).

(The abbreviation "br" means "broad").

FTIR νmax (KBr disc) 3290; 2970; 2930; 1643; 1540; 700 cm-1.

Found: C, 72.60; H, 8.02; N, 7.31.

C34H43N3O4 • 0.25 H2O requirements: C, 72.63; H, 7.80; N, 7.47%.

Example 4

N1-[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]-(2R)-2-[3-(3-fluoro-4-phenoxyphenyl) )propyl]-(N4-hydroxy)butanediamide

[image]

N-[(dimethylamino)-1H-1,2,3-triazolo[4,5-b]pyridin-1-ylmethylene]-N-methylmethaninium hexafluorophosphate N-oxide (740 mg, 1.95 mmol) was added to the stirred solution (3R)-3-({[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]amino}carbonyl)-6-(3-fluoro- 4-phenoxyphenyl)hexanoic acid (preparation 5) (731 mg, 1.30 mmol) and diisopropylethylamine (220 µl, 1.30 mmol) in anhydrous dimethylformamide (10 ml) at 0 °C under nitrogen. After 25 minutes, hydroxylamine hydrochloride (271 mg, 3.90 mmol) was added, followed by diisopropylethylamine (880 µl, 5.20 mmol). The resulting mixture was stirred at 20 °C for 16 hours, poured into ethyl acetate (150 ml) and washed with pH 7 phosphate buffer solution (3 × 50 ml), saturated aqueous sodium chloride solution (50 ml), dried (Na2SO4) and concentrated under reduced pressure. The residue was recrystallized from dichloromethane/diisopropyl ether to give the title compound as a colorless solid (275 mg, 24 %). By further purification of one sample (130 mg) by preparative HPLC (Phenomenex C18 Magellan column, 150 × 20 mm, with 5 μ packing, 20 ml acetonitrile:aqueous phosphate buffer solution (8.3 mM, pH 7.2) = 1:1 , holding time 13.5 minutes), and then by trituration with diisopropyl ether, filtering and drying in vacuum at 50 °C, a white solid was obtained.

T.t. 136-137 °C

Rf 0.39 (dichloromethane:methanol:concentrated aqueous ammonia solution = 9:10:1).

δH (400 MHz, CD3OD) 1.02 (9H, s); 1.42 (3H, d, J=7 Hz); 1.48 (3H, m); 1.58 (1H, m); 2.16 (1H, dd, J=6 and 14 Hz); 2.34 (1H, dd, J=8 and 14 Hz); 2.49 (2H, m); 2.86 (1H, m); 4.34 (1H, s); 4.99 (1H, q, J=7 Hz); 6.86 (4H, m); 6.94 (1H, d, J=11.5 Hz); 7.04 (1H, t, 7 Hz); 7.12 (1H, t, J=7 Hz); 7.18 (2H, t, J=7 Hz); 7.27 (4H, m).

LRMS (thermospray) m/z = 535 (MH+ - HNCO).

FTIR νmax (KBr disc) 3290; 2970; 2930; 1644; 1508; 1490; 1220; 700 cm-1.

Found: C, 68.14; H, 6.94; N, 7.21.

C33H40FN3O5 • 0.25 H2O requirements: C, 68.08; H, 7.01; N, 7.22%.

Example 5

N1-[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]-(N4-hydroxy)-(2R)-2-[3-(3 -methyl-4-phenoxyphenyl)propyl]butanediamide

[image]

N-[(dimethylamino)-1H-1,2,3-triazolo[4,5-b]pyridin-1-ylmethylene]-N-methylmethaninium hexafluorophosphate N-oxide (655 mg, 1.72 mmol) was added to the stirred solution (3R)-3-({[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]amino}carbonyl)-6-(3-methyl- 4-phenoxyphenyl)hexanoic acid (preparation 6) (640 mg, 1.15 mmol) and diisopropylethylamine (198 µl, 1.15 mmol) in anhydrous dimethylformamide (5 ml) at 20 °C under nitrogen. After 25 minutes, hydroxylamine hydrochloride (239 mg, 3.44 mmol) was added followed by diisopropylethylamine (792 µl, 4.60 mmol). The resulting mixture was stirred at 20 °C for 48 hours, poured into ethyl acetate (150 ml) and washed with pH 7 phosphate buffer solution (3 x 50 ml), saturated aqueous sodium chloride solution (50 ml), dried (MgSO4) and condensed under reduced pressure. The crude product was purified by flash chromatography. First, with dichloromethane:methanol:concentrated aqueous solution = 90:10:1, a yellow oil was obtained. Further purification by flash chromatography (elution with dichloromethane:methanol = 95:5) gave an orange oil, which was crystallized from dichloromethane/diisopropyl ether to give the title compound (25 mg, 4 %).

Rf 0.27 (dichloromethane: methanol = 90:10).

δH (300 MHz, CDCl 3 ) 0.98 (9H, s); 1.31 (3H, d, J=7 Hz); 1.60 (4H, m); 2.16 (3H, s); 2.24 (1H, m); 2.47 (3H, m); 2.82 (1H, m); 4.17 (1H, d, J=9 Hz); 5.05 (1H, pentet, J=7 Hz); 6.31 (1H, no d); 6.77 (1H, d, J=8 Hz); 6.85 (2H, d, J=8 Hz); 6.97 (1H, s); 7.02 (1H, t, J=8 Hz); 7.22 (9H, complex); 9.50 (1H, no. s).

LRMS (thermospray) m/z = 573 (M+, weak); 531(M+ - O).

Example 6

N1-[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]-(3S)-ethoxy-(2R)-2-[3-(3 -fluoro-4-phenoxyphenyl)propyl]-(N4-hydroxy)butanediamide

[image]

N-[(dimethylamino)-1H-1,2,3-triazolo[4,5-b]pyridin-1-ylmethylene]-N-methylmethaninium hexafluorophosphate N-oxide (88 mg, 0.232 mmol) was added to a stirred solution (2S ,3R)-3-([(1S)-2,2-dimethyl-1-([(1R)-1-phenylethyl]aminocarbonyl)propyl]aminocarbonyl)-2-ethoxy-6-(3-fluoro-4- phenoxyphenyl)hexanoic acid (preparation 8) (94 mg, 0.155 mmol) and diisopropylethylamine (54 µl, 0.31 mmol) in anhydrous dimethylformamide (2.5 ml) at 0 °C under nitrogen. After 45 minutes, hydroxylamine hydrochloride (32 mg, 0.465 mmol) was added, followed by diisopropylethylamine (81 µl, 0.465 mmol). The resulting mixture was stirred at 0 °C for 5 h, poured into ethyl acetate (50 ml) and washed with pH 7 phosphate buffered saline (3 x 30 ml), saturated aqueous sodium chloride solution (30 ml), dried ( MgSO4) and condensed under reduced pressure. The residue was purified by flash chromatography (eluting with dichloromethane:methanol:concentrated aqueous ammonia = 95:5:0.5), and the title compound was obtained as a colorless foam (43 mg, 34 %).

Rf 0.32 (dichloromethane:methanol:concentrated aqueous ammonia = 95:5:0.5).

δH (400 MHz, CD3OD) 1.04 (9H, s); 1.12 (3H, t, J=7 Hz); 1.32 (1H, m); 1.42 (3H, d, J=7 Hz); 1.46 (2H, m); 1.53 (1H, m); 2.42 (1H, dt, J=14 and 7 Hz); 2.54 (1H, ddd, J=7.9 and 14 Hz); 2.74 (1H, dt, J=10 and 7.5 Hz); 3.32 (1H, m, partially shadowed by CHD2OD peak), 3.49 (1H, dq, J=10 and 7 Hz); 3.69 (1H, d, J=10 Hz); 4.39 (1H, s); 5.00 (1H, q, J=7 Hz); 6.84 (4H, complex); 6.92 (1H, dd, J=12 and 2 Hz); 7.04 (1H, t, J=8 Hz); 7.14 (3H, complex); 7.26 (4H, complex).

LRMS (thermospray) m/z = 606 (MH+ - O); 533 (base peak, MNa+ - (Ph(CH3)CHNH2)).

HRMS (positive ion electrospray)

Found: m/z = 644.3113.

C35H44FNaN3O6 requirements: 644.3112.

Found: C, 66.79; H, 7.05; N, 6.66.

C35H44FN3O6 • 0.5 H2O requirements: C, 66.65; H, 7.17; N, 6.66%.

Example 7

(2S,3R)-3-({[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]amino}carbonyl)-6-[(3 -methyl-4-phenyl)phenyl]-2-propylhexanoic acid

[image]

According to the procedure from example 1, tert-butyl (2S,3R)-3-({[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]amino }carbonyl)-6-[3-methyl-(4-phenyl)phenyl]-2-propylhexanoate (preparation 13) (360 mg, 0.56 mmol) was treated with trifluoroacetic acid in anhydrous dichloromethane to give the title compound in as a colorless solid (163 mg, 50%) after recrystallization from methanol. The starting liquids gave a second yield of crystals (45 mg, 14%) from methanol.

T.t. 212-215 °C (from methanol).

Rf 0.4 (dichloromethane/methanol/acetic acid = 90:10:1).

δH (400 MHz, DMSO-d6) 0.77 (3H, t, J=6 Hz); 0.92 (9H, s); 1.13 (1H, m); 1.29 (3H, d, J=6.5 Hz, and 3H, m, overlap); 1.61 (4H, m); 2.13 (3H, s); 2.35 (2H, m); 2.45 (1H, m); 2.66 (1H, no t, J=8 Hz); 2.60 (1H, dd, J=9 and 15 Hz); 4.34 (1H, d, J=9 Hz); 4.90 (1H, pentet, J=6.5 Hz); 6.89 (1H, d, J=7 Hz); 6.97 (2H, s and d, J=8 Hz, overlapping); 7.25 (7H, complex); 7.31 (1H, t, J=7 Hz); 7.39 (2H, m); 7.94 (1H, no d); 8.29 (1H, no d); 11.66 (1H, no s).

LRMS (thermospray) m/z = 586 (MH+).

FTIR νmax (KBr disc) 1710; 1630 cm-1.

Found: C, 72.60; H, 8.14; N, 4.59;

C37H48N2O4 • 1.25 H2O requirements: C, 72.62, H, 8.45; N, 4.54%.

Example 8

N1-[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino)carbonyl)propyl]-(N4-hydroxy)-(2R)-2-{3-[3 -methyl-(4-phenyl)phenyl]propyl}(3S)-propylbutanediamide

[image]

According to the procedure from example 4, by the reaction (2S,3R)-3-({[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]amino}carbonyl )-6-[(3-methyl-4-phenyl)phenyl]-2-propylhexanoic acid (Example 7) (130 mg, 0.22 mmol) with hydroxylamine hydrochloride (45.9 mg, 0.66 mmol) at 20 °C for 5 hours, followed by the same treatment and trituration of the crude product with ether, the title compound (64 mg, 49%) was obtained as a colorless solid.

Rf 0.31 (dichloromethane:methanol:concentrated aqueous ammonia solution = 90:10:1).

δH (400 MHz, DMSO-d6) 0.74 (3H, t, J=6.5 Hz); 0.92 (9H, s); 1.03 (1H, m); 1.16 (2H, m); 1.24 (3H, d, J=7.5 Hz, and 1H, m, overlap); 1.37 (3H, m); 1.44 (1H, m); 2.12 (3H, s, and 1H, m, overlap); 2.33 (1H, m); 2.44 (1H, m); 2.61 (1H, m); 4.31 (1H, d, J=9 Hz); 4.89 (1H, pentet, J=7.5 Hz); 6.88 (1H, d, J=8 Hz); 6.95 (1H, s); 6.97 (1H, d, J=8 Hz); 7.13 (3H, m); 7.22 (4H, m); 7.30 (1H, t, J=8 Hz); 7.38 (2H, t, J=8 Hz); 7.88 (1H, no d); 8.18 (1H, no d); 8.73 (1H, no s); 10.28 (1H, no s).

LRMS (thermospray) m/z (base peak, MNH4+ - HCONHOH).

Found: C, 72.14; H, 8.18; N, 6.85.

C37H49N3O4 • 0.75 H2O requirements: C, 72.46; H, 8.30; N, 6.85%.

FTIR νmax (KBr disc) 3290; 2970; 2930; 1637; 1530; 700 cm-1.

Example 9

(N4,3S)-dihydroxy-N1-[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]-(2R)-2-[3- (3-fluoro-4-phenoxyphenyl)propyl]butanediamide

[image]

Sodium methoxide (35 mg, 0.67 mmol) was added to a solution of anhydrous hydroxylamine hydrochloride (45 mg, 0.67 mmol) in anhydrous methanol (1 mL) under nitrogen at room temperature. The mixture was stirred for 2 hours and filtered rapidly through a piece of Arbocel filter aid, washing with anhydrous methanol (1 ml). (2R)-2-[(4S)-2,2-dimethyl-5-oxo-1,3-dioxolan-4-yl]-N-[(1S)-2,2-dimethyl-1-( {[(1R)-1-phenylethyl]amino}carbonyl)propyl]-5-[(3-fluoro-4-phenoxy)phenyl]pentanamide (preparation 15) (103 mg, 0.16 mmol) and anhydrous methanol (1 ml), so the mixture was stirred at room temperature for 18 hours. The solution was concentrated under reduced pressure, and the residue was purified by flash chromatography, first over C18 silanized silica gel (40-63 µ) eluting with methanol:water = 4:1, and then over ordinary silica gel (gradient elution with dichloromethane:methanol). The product was obtained as a colorless solid (25 mg, 26 %).

T.t. 90-95 °C.

Rf 0.38 (dichloromethane:methanol = 90:10).

δH (300 MHz, CD3OD) 1.03 (9H, s); 1.45 (3H, d, J=7 Hz); 1.52 (3H, m); 1.66 (1H, m); 2.53 (2H, m); 2.77 (1H, m); 4.02 (1H, d, J=8 Hz); 4.36 (1H, s); 5.01 (1H, q, J=7 Hz); 6.87 (4H, complex); 6.97 (1H, d, J=12 Hz); 7.04 (1H, t, J=8 Hz); 7.16 (3H, complex); 7.28 (4H, complex).

LRMS (thermospray) m/z (base peak, MH+ - HONHCO).

Found: C, 65.94; H, 6.89; N, 6.96.

C33H40FN3O6 • 0.4 H2O requirements: C, 65.96; H, 6.84; N, 6.99%.

Example 10

(N4,3S)-dihydroxy-N1-[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]-(2R)-2-{3- [3-Methyl-(4-phenyl)phenyl]propyl}butanediamide

[image]

According to the procedure from example 9, (2R)-2-[(4S)-2,2-dimethyl-5-oxo-1,3-dioxolan-4-yl]-N-[(1S)-2,2-dimethyl -1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]-5-[(3-methyl-4-phenyl)phenyl]pentathanamide (preparation 16) (440 mg, 0.73 mmol) added is reacted with hydroxylamine at room temperature for 18 hours. The solution was concentrated under reduced pressure, and the residue was purified by flash chromatography (C18 silanized silica gel (40-63 µ) eluting with methanol:water = 4:1 and then 5:1) to give a colorless solid (339 mg, 81 %).

T.t. 95-97 °C.

Rf (dichloromethane:methanol = 95:5).

δH (300 MHz, CDCl 3 ) 0.98 (9H, s); 1.50 (3H, d, J=7 Hz); 1.65 (3H, m); 1.89 (1H, m); 2.23 (3H, s); 2.58 (2H, br t, J=6 Hz); 3.27 (1H, m); 4.15 (1H, d); 4.20 (1H, no s); 5.09 (1H, pentet, J=7 Hz); 5.32 (1H, no s); 6.20 (1H, no d); 6.98 (1H, d, J=8 Hz); 7.01 (1H, s); 7.10 (1H, d, J=8 Hz); 7.31 (10H, complex); 7.86 (number d); 9.50 (1H, no. s).

LRMS (thermospray) m/z = 514 (base peak, MH+ - HONHCO).

Found: C, 70.50; H, 7.58; N, 7.23.

C34H43N3O5 • 0.25 H2O requirements: C, 70.62; H, 7.58; N, 7.27%.

Example 11

(2R)-N1-[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]-2-{3-[3-fluoro-(4- phenyl)phenyl]propyl}-(N4-hydroxy)butanediamide

[image]

a) According to the procedure from example 2, (3R)-3-({[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]amino}carbonyl) -6-[(3-fluoro-4-phenyl)phenyl]hexanoic acid (preparation 17) (426 mg, 0.78 mmol) was reacted with O-allylhydroxylamine hydrochloride (128 mg, 1.17 mmol). Purification of the crude product by flash chromatography (gradient elution with hexane:isopropanol) followed by trituration with ether and ethyl acetate gave (2R)-N1-[(1S)-2,2-dimethyl-1-({[(1R)-1- phenylethyl]amino}carbonyl)propyl]-2-{3-[3-fluoro-(4-phenyl)phenyl]propyl}(N4-3-propenyloxy)butanediamide (260 mg, 55%) as a white solid.

T.t. 182-186 °C.

Rf 0.32 (hexane:isopropanol = 10.1).

δH (400 MHz, DMSO-d6) 0.87 (9H, s); 1.26 (3H, d, J= 6.5 Hz); 1.28 (1H, m); 1.43 (3H, m); 1.97 (1H, m); 2.13 (1H, m); 2.50 (2H, m); 2.84 (1H, m); 4.16 (2H, m); 4.27 (1H, d, J= 9 Hz); 4.87 (1H, pentet, J= 6.5 Hz); 5.15 (1H, d, J= 10 Hz); 5.20 (1H, d, J= 17 Hz); 5.85 (1H, m); 6.97 (2H, m); 7.13 (4H, complex); 7.29 (2H, m); 7.45 (4H, m); 7.65 (1H, no s); 8.26 (1H, no s); 10.37 (1H, no s).

LRMS (thermospray) m/z = 602 (MH+).

Found: C, 70.91; H, 7.33; N, 6.84.

C36H44FN3O4 • 0.5 H2O requirements: C, 70.80; H, 7.43; N, 6.88%.

b) According to the procedure from example 2, (2R)-N1-[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]-2-{3- [3-Fluoro-(4-phenyl)phenyl]propyl}-(N4-3-propenyloxy)butanediamide (210 mg, 0.35 mmol) was reacted with ammonium formate (110 mg, 1.75 mmol) in ethanol :water (4:1, 5 ml) with palladium catalysis at reflux for 2 hours. After treatment, the residue was purified by flash chromatography (gradient elution dichloromethane:methanol:concentrated aqueous ammonia solution) and trituration with ether, (2R)-N1-[(1S)-2,2-dimethyl-1-({[(1R )-1-phenylethyl]amino}carbonyl)propyl]-2-{3-[3-fluoro-(4-phenyl)phenyl]propyl}(N4-hydroxy)butanediamide (120 mg, 61 %) as a colorless solid matter.

Rf 0.25 (dichloromethane:methanol:concentrated aqueous ammonia solution = 90:10:1)

δH (400 MHz, DMSO-d6) 0.87 (9H, s); 1.25 (3H, d, J= 6.5 Hz); 1.31 (1H, m); 1.44 (3H, m); 2.00 (1H, dd, J= 7 and 15 Hz); 2.13 (1H, dd, J= 4 and 15 Hz); 2.48 (2H, m); 2.80 (1H, m); 4.26 (1H, d, J= 9 Hz); 4.87 (1H, pentet, J= 6.5 Hz); 6.97 (2H, m); 7.06 (1H, m); 7.13 (2H, t, J= 8 Hz); 7.20 (2H, m); 7.30 (2H, m); 7.43 (4H, complex); 7.66 (1H, no d); 8.23 (1H, no d); 8.58 (1H, s); 10.27 (1H, no s).

LRMS (thermospray) m/z = 563 (MH+).

Found: C, 70.02; H, 7.25; N, 7.52.

C33H40FN3O4 • 0.25 H2O requirements: C, 70.00; H, 7.21; N, 7.42%.

Example 12

(3R)-3-({[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]amino}carbonyl)-(2S)-ethoxy-6 -[(3-methyl-4-phenyl)phenyl]hexanoic acid

[image]

Lithium hydroxide hydrate (30 mg, 0.71 mmol) was added to a suspension of methyl (2S,3R)-3-({[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl) amino}carbonyl)propyl]amino}carbonyl)-2-ethoxy-6-[(3-methyl-4-phenyl)phenyl]hexanoate (preparation 18) (384 mg, 0.64 mmol) in tetrahydrofuran:water = 3: 2 (10 ml), and the mixture was stirred at room temperature for 2 hours. The solution was acidified with 1M hydrochloric acid and extracted with ethyl acetate (3 × 30 ml). The combined extracts were dried (Na2SO4) and concentrated under reduced pressure. Trituration with ether gave the title compound (196 mg, 52%) as a colorless solid.

δH (400 MHz, CD3OD) (substitutable hydrogens only partially substituted) 1.02 (9H, s); 1.16 (3H, t, J = 7 Hz); 1.42 (3H, d, J = 7 Hz); 1.52 (3H, m); 1.65 (1H, m); 2.16 (3H, s); 2.50 (2H, m); 2.74 (1H, m); 3.39 (1H, m); 3.58 (1H, m); 3.90 (1H, d, J= 9 Hz); 4.38 (1H, d, J= 9 Hz); 5.00 (1H, pentet, J= 7 Hz); 6.92 (2H, m); 7.00 (2H, m); 7.20 (7H, complex); 7.38 (2H, t, J=7); 7.86 (1H, no d); 8.35 (1H, no. d).

Found: C, 73.11; H, 7.89; N, 4.79.

C36H46N2O5 • 0.25 H2O requirements: C, 73.12; H, 7.92; N, 4.74%.

Example 13

N1-[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]-(3S)-ethoxy-(N4-hydroxy)-(2R)-2 -{3-[3-methyl-(4-phenyl)phenyl]propyl}butanediamide

[image]

N-[(dimethylamino)-1H-1,2,3-triazolo[4,5-b]pyridin-1-ylmethylene]-N-methylmethaninium hexafluorophosphate N-oxide (164 mg, 0.43 mmol) was added to the stirred solution (2S,3R)-3-({[(1S)-2,2-dimethyl-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]amino}carbonyl)-2-ethoxy-6-[ (3-methyl-4-phenyl)phenyl]hexanoic acid (Example 12) (169 mg, 0.29 mmol) and diisopropylethylamine (50 µl, 0.29 mmol) in anhydrous dimethylformamide (3 ml) at 0 °C under nitrogen . After 60 minutes, hydroxylamine hydrochloride (60 mg, 0.86 mmol) was added, followed by diisopropylethylamine (81 µl, 0.465 mmol). The resulting mixture was stirred at 20 °C for 16 hours, diluted with ether and washed with water. The aqueous layer was extracted with ether (2×) and ethyl acetate. The combined organic extracts were washed with water, dried (Na2SO4) and concentrated under reduced pressure. The residue was purified by repeated flash chromatography (eluting with dichloromethane:methanol:concentrated aqueous ammonia solution = 97.5:2.5:0.5, then 96:4:0.5) and the title compound was obtained as a colorless foam ( 20 mg, 11 %).

Rf 0.25 (dichloromethane:methanol:concentrated aqueous ammonia solution = 90:10:1).

δH (300 MHz, CDCl 3 ) 0.97 (9H, s); 1.31 (3H, t, J = 7 Hz); 1.55 (3H, d, J= 7 Hz); 1.71 (3H, m); 1.84 (1H, m); 2.23 (3H, s); 2.65 (2H, m); 2.84 (1H, m); 3.48 (1H, pentet, J = 7 Hz); 3.76 (1H, m); 3.95 (1H, d, J= 7 Hz); 3.98 (1H, d, J= 10 Hz); 5.12 (1H, q, J = 7 Hz); 6.47 (1H, no d); 7.02 (2H, m); 7.13 (1H, d, J= 8 Hz); 7.30 (10H, complex); 9.65 (1H, no s).

HRMS (positive ion electrospray)

Found: m/z = 624.3403.

C36H47NaN3O5 requires 624.3413.

Example 14

(3R)-3-({[(1S)-2,2-dimethyl-1-({[(1S)-2-methoxy-1-phenylethyl]amino}carbonyl)propyl]amino}carbonyl)-6-[ (3-methyl-4-phenyl)phenyl]hexanoic acid

[image]

According to the procedure from example 1, tert-butyl (3R)-3-({[(1S)-2,2-dimethyl-1-({[(1S)-2-methoxy-1-phenylethyl]amino}carbonyl)propyl ]amino}carbonyl)-6-[3-methyl-(4-phenyl)phenyl]-hexanoate (preparation 19) (535 mg, 0.85 mmol) was treated with trifluoroacetic acid in anhydrous dichloromethane at room temperature for 2 h. The residue was dissolved in toluene and concentrated under reduced pressure (2×), then recrystallized from ethyl acetate to give a colorless solid (387 mg, 80%).

T.t. 184-186 °C (from ethyl acetate).

Rf 0.47 (hexane/ether/acetic acid = 50:50:1).

δH (400 MHz, CD3OD) (substitutable hydrogens only partially substituted) 1.03 (9H, s); 1.51 (4H, m); 2.15 (3H, s); 2.36 (1H, dd, J= 5 and 17 Hz); 2.46 (2H, m); 2.60 (1H, dd, J= 10 and 17 Hz); 2.82 (1H, m); 3.32 (3H, s); 3.57 (2H, d, J = 7 Hz); 4.42 (1H, d, J= 10 Hz); 5.10 (1H, q, J = 7 Hz); 6.87 (1H, d, J = 8 Hz); 6.98 (2H, s and d, J= 8 Hz, overlapping); 7.22 (8H, complex); 7.39 (2H, t, J = 7 Hz); 7.74 (1H, no d); 8.48 (1H, no. d).

LRMS (thermospray) m/z = 573 (MH+).

FTIR νmax (KBr disc) 3300; 2960; 2930; 1711; 1639; 1543; 700 cm-1.

Found: C, 73.32; H, 7.73; N, 4.80.

C35H44N2O5 requirements: C, 73.40; H, 7.74; N, 4.89%.

Example 15

(3R)-3-({[(1S)-2,2-dimethyl-1-({[(1S)-2-methoxy-1-phenylethyl]amino}carbonyl)propyl]amino}carbonyl)-6-( 3'-Methoxy-2-methylbiphen-4-yl)hexanoic acid

[image]

According to the procedure from example 1 tert-butyl (3R)-3-({[(1S)-2,2-dimethyl-1-({[(1S)-2-methoxy-1-phenylethyl]amino}carbonyl)propyl] amino}carbonyl)-6-(3'-methoxy-2-methylbiphen-4-yl)hexanoate (preparation 21) (660 mg, 1.0 mmol) was treated with trifluoroacetic acid in anhydrous dichloromethane at room temperature for 3 hours. The residue was dissolved in toluene and concentrated under reduced pressure (2×) then recrystallized from ethyl acetate to give a colorless solid (326 mg, 54%). Recrystallization of the original liquid gave another 107 mg (18 %).

T.t. 155.5-157.5 °C (from ethyl acetate).

Rf 0.34 (hexane/ether/acetic acid = 50.50:1).

δH (400 MHz, CD3OD) (substitutable hydrogens only partially substituted) 1.03 (9H, s); 1.51 (4H, m); 2.16 (3H, s); 2.37 (1H, dd, J= 5 and 17 Hz); 2.47 (2H, m); 2.59 (1H, dd, J= 10 and 17 Hz); 2.83 (1H, m); 3.32 (3H, s); 3.57 (2H, d, J = 7 Hz); 3.80 (3H, s); 4.43 (1H, d, J= 10 Hz); 5.10 (1H, q, J = 7 Hz); 6.77 (1H, s); 6.80 (1H, d, J= 7 Hz); 6.87 (2H, m); 6.97 (2H, m); 7.16 (3H, m); 7.28 (3H, m); 7.74 (1H, no d); 8.50 (1H, no. d).

LRMS (thermospray) m/z 603 (MH+).

FTIR νmax (KBr disc) 3300; 2960; 2930; 1711; 1640; 1545; 1484; 1217; 1211; 700 cm-1.

Found: C, 71.77; H, 7.69; N, 4.61.

C36H46N2O6 required: C, 71.73; H, 7.69; N, 4.65%.

Example 16

(2R)-N-1-[(1S)-2,2-dimethyl-1-({[(1S)-2-methoxy-1-phenylethyl]amino}carbonyl)propyl]-2-[(3-[ (3-Methyl-4-phenyl)phenyl]propyl}-(N-4-hydroxy)butanediamide

[image]

a) According to the procedure from example 2, (3R)-3-({[(1S)-2,2-dimethyl-1-({[(1S)-2-methoxy-1-phenylethyl]amino}carbonyl)propyl] amino}carbonyl)-6-[(3-methyl-4-phenyl)phenyl]hexanoic acid (Example 14) (347 mg, 0.61 mmol) was reacted with O-allylhydroxylamine hydrochloride (81 mg, 0.73 mmol). Purification of the crude product by flash chromatography (eluting with hexane:ethyl acetate = 2:1), followed by trituration with ether and ethyl acetate, yielded (2R)-N1-[(1S)-2,2-dimethyl-1-({[ (1S)-2-methoxy-1-phenylethyl]amino}carbonyl)propyl]-2-{3-[(3-methyl-4-phenyl)phenyl]propyl}-(N4-3-propenyloxy)butanediamide (306 mg , 80%) in the form of solid white matter.

T.t. 117-120 °C.

Rf 0.28 (hexane:ethyl acetate = 1:2).

δH (400 MHz, CDCl 3 ) 1.02 (9H, s); 1.40-1.70 (4H, complex); 2.20 (3H, s, and 1H, m, overlap); 2.34-2.60 (3H, complex); 2.76 (1H, m); 3.36 (3H, s); 3.63 (2H, d, J = 5 Hz); 4.28 (1H, d, J = 9.5 Hz); 4.34 (2H, d, J = 6 Hz); 5.12 (1H, dt, J= 7.5 and 5 Hz); 5.30 (2H, m); 5.90 (1H, m); 6.44 (1H, d, J= 7.5 Hz and 1H, br s, overlap); 6.94 (1H, m); 6.98 (1H, s); 7.08 (1H, d, J= 7 Hz); 7.18-7.38 (8H, complex); 7.40 (2H, m); 8.50 (1H, no. s).

LRMS (thermospray) m/z = 628 (MH+).

b) Mixed mixture (2R)-N1-[(1S)-2,2-dimethyl-1-({[(1S)-2-methoxy-1-phenylethyl]amino}carbonyl)propyl]-2-{3- [(3-methyl-4-phenyl)phenyl]propyl}-(N4-3-propenyloxy)butanediamide (300 mg, 0.48 mmol) and ammonium formate (300 mg, 4.76 mmol) were dissolved in a hot mixture of ethanol /water (4:1, 6 ml) and a colorless solution was obtained. A solution of palladium acetate (4 mg, 0.018 mmol) and triphenylphosphine (9.6 mg, 0.037 mmol) in ethanol/water (4:1, 2 ml) was added, and the mixture was heated under reflux for 60 minutes. After cooling, the brown solution was diluted with ethyl acetate (100 ml), washed with concentrated aqueous sodium chloride solution (2 x 50 ml), dried (MgSO4) and concentrated under reduced pressure. The residue was purified by column chromatography (C18 silanized silica gel (40-63 µ), eluting with methanol:water = 4:1), and then by trituration with diisopropyl ether, (2R)-N1-[(1S)-2,2 was obtained -dimethyl-1-({[(1S)-2-methoxy-1-phenylethyl]amino}carbonyl)propyl]-2-{3-[(3-methyl-4-phenyl)phenyl]propyl)-(N4- hydroxy) butanediamide (226 mg, 80%) as a white solid.

T.t. 92-96 °C

Rf 0.57 (dichloromethane:methane:concentrated aqueous ammonia solution = 90:10:1).

δH (400 MHz, CH3OD) 1.02 (9H, s); 1.40-1.68 (4H, m); 2.17 (3H, s); 2.20 (1H, m); 2.36 (1H, m); 2.48 (2H, m); 2.86 (1H, m); 3.30 (3H, s); 3.58 (2H, d, J = 6.5 Hz); 4.40 (1H, s); 5.10 (1H, t, J= 6.5 Hz); 6.88 (1H, m); 6.98 (2H, m); 7.10-7.36 (8H, complex); 7.39 (2H, m).

LRMS (thermospray) m/z = 588 (MH+).

Found: C, 79.95; H, 7.85; N, 7.00.

C35H45N3O5 • 0.25 H2O requirements: C, 70.98; H, 7.74; N, 7.09%.

Examples 17 and 18

(N4,3S)-dihydroxy-N1-[(1S)-2,2-dimethyl-1-((1R)-1-phenylethyl]amino}carbonyl)propyl]-(2R)-2-{3-[4 -(4-cyanophenyl)-3-methylphenyl]propyl}butanediamide, and (N4,3S)-dihydroxy-N1-[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl] amino}carbonyl)propyl]-(2R)-2-{3-[4-(4-hydroxyamidino)phenyl-3-methylphenyl]propyl]propyl}butanediamide

[image]

According to the procedure from example 9, (2R)-5-{[4-(4-cyanophenyl)-3-methyl]phenyl}-2-[(4S)-2,2-dimethyl-5-oxo-1,3- dioxolan-4-yl]-N-[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]pentanamide (Preparation 22) (384 mg, 0, 62 mmol) was reacted with hydroxylamine at room temperature for 18 hours. The solution was concentrated under reduced pressure, and the residue was purified by column chromatography (C18 silanized silica gel (40-63 µ) eluting with methanol:water = 70:30 and then 80:20) and two fractions were obtained. The first eluted product was identified as (N4,3S)-dihydroxy-N1-[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]-(2R) -2-{3-[4-(4-cyanophenyl)-3-methylphenyl]propyl}butanediamide, Rf 0.38 (dichloromethane:methanol = 90:10), which was triturated with diisopropyl ether to give a white solid (115 mg , 31 %).

T.t. 103-109 °C.

δH (400 MHz, DMSO-d6) 0.92 (9H, s); 1.25 (1H, m); 1.26 (3H, d, J = 7.5 Hz); 1.40-1.54 (3H, m); 2.14 (3H, s); 2.40 (1H, m); 2.44 (1H, m); 2.70 (1H, m); 3.76 (1H, t, J = 7.5 Hz); 4.34 (1H, d, J = 9.5 Hz); 4.90 (1H, pentet, J= 7.5 Hz); 5.24 (1H, d, J = 7.5 Hz); 6.98 (1H, m); 7.00 (2H, m); 7.10 (3H, m); 7.20 (2H, m); 7.46 (2H, d, J = 8 Hz); 7.58 (1H, d, J = 9.5 Hz); 7.82 (2H, d, J = 8 Hz); 8.26 (1H, d, J = 7.5 Hz); 8.80 (1H, no s); 10.56 (1H, no s).

LRMS (thermospray) m/z = 538 (base peak, M+ - HONHCO).

Found: C, 69.26; H, 7.15; N, 9,10.

C35H42N4O5 • 0.1 EtOAc • 0.4 H2O found: C, 69.16; H, 7.15; N, 9.11%.

The second eluted product was identified as (N4,3S)-dihydroxy-N1-[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]-(2R) -2-{3-[4-(4-hydroxamidino)-phenyl-3-methylphenyl]propyl}butanediamide,

Rf 0.20 (dichloromethane:methanol = 90:10) as a white solid (84 mg, 21 %).

T.t. 131-135 °C.

δH (400 MHz, DMSO-d6) 0.90 (9H, s); 1.24 (1H, m, and 3H, d, J= 7 Hz, overlap); 1.38-1.58 (3H, m); 2.16 (3H, s); 2.40 (1H, m); 2.44 (1H, m); 2.58 (1H, m); 3.76 (1H, t, J = 7 Hz); 4.34 (1H, d, J= 10.5 Hz); 4.90 (1H, pentet, J= 7 Hz); 5.24 (1H, d, J = 7 Hz); 5.80 (2H, s); 6.96 (1H, m); 7.00 (2H, m); 7.04-7.20 (3H, m); 7.21 (2H, m); 7.46 (2H, d, J = 8 Hz); 7.58 (1H, d, J = 10.5 Hz); 7.70 (2H, d, J= 8 Hz); 8.24 (1H, d, J= 7 Hz); 8.80 (1H, no s); 9.60 (1H, s); 10.58 (1H, no s).

LRMS (thermospray) m/z = 632 (MH+).

Found: C, 64.86; H, 7.21; N, 10,28.

C35H45N5O6 • 0.1 CH3OH • 0.9 H2O finds: C, 64.74; H, 7.31; N, 10.75%.

Example 19

(N4,3S)-dihydroxy-N1-[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]-(2R)-2-{3- [4-(3-Cyanophenyl)-3-methylphenyl]propyl}butanediamide

[image]

Sodium methoxide (43 mg, 0.80 mmol) was added to a solution of anhydrous hydroxylamine hydrochloride (56 mg, 0.80 mmol) in anhydrous methanol (1 mL) under nitrogen at room temperature. The mixture was stirred for 2.5 hours and rapidly filtered through a piece of Arbocel filter aid, washing with anhydrous methanol (1 ml). (2R)-5-{[4-(3-cyanophenyl)-3-methyl]phenyl}-2-[(4S)-2,2-dimethyl-5-oxo-1,3-dioxolan-4-yl] -N-[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]pentanamide (preparation 24) (123 mg, 0.20 mmol) in anhydrous methanol (1 ml) was added to the mixture at 4 °C and stirred for 15 min. The mixture was heated to room temperature and stirred for 9 hours. The solution was concentrated under reduced pressure and the residue was purified by column chromatography (C18 silanized silica gel (40-63 µ) eluting with methanol:water = 3:1). The residue was azeotroped with ethanol and then ethyl acetate and triturated with diisopropyl ether to give the title compound as a non-crystalline white solid (58 mg, 48 %).

T.t. has not been determined.

Rf 0.11 (dichloromethane:methanol:concentrated aqueous ammonia solution = 90:10:1).

δH (400 MHz, DMSO-d6) 0.92 (9H, s); 1.24 (1H, m, and 3H, d, J= 7 Hz, overlap); 1.38-1.54 (3H, m); 2.14 (3H, s); 2.40 (1H, m); 2.48 (1H, m); 2.70 (1H, m); 3.76 (1H, t, J = 7.5 Hz); 4.34 (1H, d, J = 9.5 Hz); 4.90 (1H, pentet, J= 7 Hz); 5.22 (1H, d, J = 7.5 Hz); 6.98 (1H, d, J= 7 Hz); 7.02 (2H, m); 7.14 (3H, m); 7.22 (2H, d, J = 7 Hz); 7.58 (1H, d, J = 9.5 Hz); 7.60 (2H, d, J= 5 Hz); 7.72 (1H, s); 7.80 (1H, m); 8.26 (1H, d, J = 7 Hz); 8.80 (1H, no s); 10.56 (1H, no s).

LRMS (thermospray) m/z = 509 (MH+ (base peak, M+ - HONHCO).

Found: C, 69.12; H, 7.20; N, 9.04.

C35H42N4O5 • 0.5 H2O requirements: C, 69.17; H, 7.13; N, 9.22%.

Example 20

(2R)-2-{3-[4-(3-carbamoylphenyl)-3-methylphenyl]propyl}-(N4,3S)-dihydroxy-N1-[(1S)-2,2-dimethyl-1-({ [(1R)-1-phenylethyl]amino}carbonyl)propyl]butanediamide

[image]

According to the procedure from example 19, (2R)-5-{[4-(3-carbamoylphenyl)-3-methyl]phenyl-2-[(4S)-2,2-dimethyl-5-oxo-1,3-dioxolane -4-yl]-N-[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]pentanamide (preparation 26) (126 mg, 0.20 mmol) was reacted with hydroxylamine at room temperature for 13 hours. The solution was concentrated under reduced pressure, and the residue was purified by column chromatography (C18 silanized silica gel (40-63 µ) eluting with methanol:water = 3:2). The residue was azeotroped with ethanol, then ethyl acetate and triturated with diethyl ether, after which the title compound was obtained as a white solid (74 mg, 60%).

T.t. 85-100 °C.

Rf 0.04 (dichloromethane:methanol:concentrated aqueous ammonia solution = 90:10:1).

δH (400 MHz, DMSO-d6) 0.94 (9H, s); 1.24 (1H, m, and 3H, d, J= 7 Hz, overlap); 1.43 (3H, m); 2.14 (3H, s); 2.40 (1H, m); 2.50 (1H, m); 2.70 (1H, m); 3.76 (1H, t, J = 7 Hz); 4.36 (1H, d, J= 10 Hz); 4.90 (1H, pentet, J= 7 Hz); 5.24 (1H, d, J = 7.5 Hz); 6.98 (1H, d, J= 7 Hz); 7.02 (2H, m); 7.14 (3H, m); 7.20 (2H, m); 7.34 (1H, s); 7.40 (1H, d, J= 7 Hz); 7.46 (1H, d, J = 7 Hz); 7.58 (1H, d, J= 10 Hz); 7.80 (1H, s); 7.82 (1H, d, J = 7 Hz); 7.98 (1H, s); 8.26 (1H, d, J = 7 Hz); 8.90 (1H, s); 10.60 (1H, s).

LRMS (thermospray) m/z = 617 (MH+).

Found: C, 65.35; H, 7.29; N, 8.57.

C35H44N4O6 • 1.33 H2O requirements: C, 65.61; H, 7.34; N, 8.74%.

Example 21

(3R)-3-[({({[(1S)-2-methoxy-1-phenylethyl]amino}carbonyl)-[(1S)-2-methyl]-1-propyl}amino)carbonyl]-6- [3-Methyl-(4-phenyl)phenyl]hexanoic acid

[image]

According to the procedure from example 1, tert-butyl (3R)-3-[({({[(1S)-2-methoxy-1-phenylethyl]amino}carbonyl)-[(1S)-2-methyl]-1- propyl}amino)carbonyl]-6-[3-methyl-(4-phenyl)phenyl]hexanoate (preparation 28) (700 mg, 1.14 mmol) was treated with trifluoroacetic acid in anhydrous dichloromethane at room temperature for 4 hours. The residue was azeotroped with toluene (2×), and recrystallization from diethyl ether gave the title compound as a colorless solid (506 mg, 79 %).

T.t. 141-144 °C.

Rf 0.28 (pentane:ethyl acetate:acetic acid = 50:50:1).

δH (400 MHz, DMSO-d6) 0.83 (3H, d, J= 8 Hz); 0.85 (3H, d, J= 8 Hz); 1.36 (1H, m); 1.48 (3H, m); 1.94 (1H, octet, J= 8 Hz); 2.18 (3H, s); 2.22 (1H, m); 2.42 (3H, m); 2.76 (1H, m); 3.20 (3H, s); 3.44 (2H, d, J = 7.5 Hz); 4.20 (1H, t, J= 8 Hz); 5.00 (1H, q, J= 7.5 Hz); 6.94 (1H, m); 7.00 (2H, m); 7.12-7.38 (8H, complex); 7.40 (2H, m); 7.90 (1H, d, J= 8 Hz); 8.02 (1H, d, J = 7.5 Hz); 12.04 (1H, no s).

LRMS (thermospray) m/z = 559 (MH+).

Found: C, 72.86; H, 7.52; N, 5.04.

C34H42N2O5 required: C, 73.09: H, 7.58; N, 5.01%.

Example 22

(3R)-3-({[({[(1S)-2-methoxy-1-phenylethyl]amino}carbonyl)-(1S)-2-phenylethyl]amino}carbonyl)-6-[3-methyl-( 4-phenyl)phenyl]hexanoic acid

[image]

According to the procedure from example 1, tert-butyl (3R)-3-({[({[(1S)-2-methoxy-1-phenylethyl]amino}carbonyl)-(1S)-2-phenylethyl]amino}carbonyl) -6-[3-methyl-(4-phenyl)phenyl]hexanoate (preparation 30) (920 mg, 1.39 mmol) was treated with trifluoroacetic acid in anhydrous dichloromethane at room temperature for 4 hours. The residue was azeotroped with toluene (2×) and recrystallized from diethyl ether to give the title compound as a colorless solid (665 mg, 79 %).

T.t. 152-157 °C.

Rf 0.27 (pentane:ethyl acetate:acetic acid = 50:50:1).

δH (400 MHz, DMSO-d6) 1.30-1.50 (4H, m); 2.16 (3H, s); 2.18 (1H, m); 2.30 (1H, m); 2.42 (2H, m); 2.62 (1H, m); 2.84 (1H, m); 2.98 (1H, m); 3.18 (3H, s); 3.38 (2H, d, J= 6 Hz); 4.60 (1H, dt, J= 6 and 8.5 Hz); 4.98 (1H, q, J= 8 Hz); 6.94 (1H, m); 7.00 (2H, m); 7.12-7.38 (13H, complex); 7.40 (2H, m); 8.08 (1H, d, J = 8.5 Hz); 8.20 (1H, d, J= 8 Hz); 12.05 (1H, no s).

LRMS (thermospray) m/z = 607 (MH+).

Found: C, 72.02; H, 6.96; N, 4.63.

C38H42N2O5 required: C, 75.22; H, 6.98; N, 4.62%.

Example 23

(3R)-3-({[(1S)-2,2-dimethyl-1-{[(1R)-1-(4-pyridyl)ethylamino]carbonyl}propyl]amino}carbonyl)-6-[(3 -methyl-4-phenyl)phenyl]hexanoic acid hydrochloride

[image]

Hydrogen chloride gas was bubbled through the solution of tert-butyl (3R)-3-({[(1S)-2,2-dimethyl-1-{[(1R)-1-(4-pyridyl)ethylamino]carbonyl}propyl]amino }carbonyl)-6-[(3-methyl-4-phenyl)phenyl]hexanoate (preparation 31) (285 mg, 0.47 mmol) in dioxane (15 ml) under nitrogen at 0 °C, until satd. The solution was stirred for 3 hours at 0 °C, and then concentrated under reduced pressure. The residue was triturated with diethyl ether to give the title compound as a white solid (390 mg, 88 %).

T.t. 132 °C (decomposition).

Rf 5 (ethyl acetate:hexane:acetic acid = 75:25:1)

δH (400 MHz, CD3OD) 1.06 (9H, s); 1.50 (4H, m, and 3H, d, J= 7 Hz, overlapping); 2.16 (3H, s); 2.40-2.60 (3H, m); 2.64 (1H, m); 2.90 (1H, m); 4.32 (1H, d, J= 10 Hz); 5.16 (1H, pentet, J = 7 Hz); 6.86 (1H, m); 6.98 (2H, m); 7.21 (2H, m); 7.30 (1H, m); 7.40 (2H, m); 7.90 (1H, d, J= 10 Hz); 7.96 (2H, d, J = 8 Hz); 8.56 (2H, d, J = 8 Hz); 8.90 (1H, d, J= 7 Hz).

LRMS (thermospray) m/z = 544 (MH+).

Found: C, 65.43; H, 7.28; N, 6.72.

C33H41N3O4 • 1.5 H2O requirements: C, 65.28; H, 7.47; N, 6.92%.

Example 24

N1-[(1S)-2,2-dimethyl-1-([(1R)-1-(3-pyridyl)ethylamino]carbonyl)propyl]-(N4-hydroxy)-(2R)-2-{3- [3-Methyl-(4-phenyl)phenyl]propyl}butanediamide

[image]

a) According to the procedure from example 2, (3R)-3-({[(1S)-2,2-dimethyl-1-{[(1R)-1-(3-pyridyl)ethylamino]carbonyl}propyl]amino} carbonyl)-6-[(3-methyl-4-phenyl)phenyl]hexanoic acid (preparation 32) (330 mg, 0.57 mmol) was reacted with O-allylhydroxyamine hydrochloride (76 mg, 0.68 mmol) . Purification of the crude product by recrystallization from hot ethyl acetate gave N1-[(1S)-2,2-dimethyl-1-([(1R)-1-(3-pyridyl)ethylamino]carbonyl)propyl]-(N4-3- propenyloxy)-(2R)-2-{3-[3-methyl-(4-phenyl)phenyl]propyl}butanediamide (219 mg, 64%) as a white solid.

T.t. 187-189 °C.

Rf 0.21 (ethyl acetate).

δH (400 MHz, DMSO-d6) 0.90 (9H, s); 1.32 (3H, d, J = 7.5 Hz); 1.42 (4H, m); 2.04 (1H, m); 2.16 (3H, s); 2.18 (1H, m); 2.46 (2H, m); 2.86 (1H, m); 4.20 (2H, m); 4.28 (1H, d, J = 9 Hz); 4.96 (1H, m); 5.20 (2H, m); 5.88 (1H, m); 6.94 (1H, m); 7.00 (2H, m); 7.18 (1H, m); 7.26 (2H, m); 7.32 (1H, m); 7.40 (2H, m); 7.60 (1H, m); 7.70 (1H, m); 8.38 (1H, m); 8.40 (1H, d, J = 7.5 Hz); 8.48 (1H, s); 10.85 (1H, s).

LRMS (APCI) m/z = 599 (MH + ).

b) According to the procedure from example 2, N1-[(1S)-2,2-dimethyl-1-([(1R)-1-(3-pyridyl)ethylamino]carbonyl)propyl]-{N4-3-propenyloxy) -(2R)-2-{3-[3-methyl-(4-phenyl)phenyl]propyl}butanediamide (219 mg, 0.37 mmol) was reacted with ammonium formate (230 mg, 3.66 mmol) in ethanol/water (4:1, 8 ml) with palladium catalysis at reflux for 1.25 hrs. After work-up, the residue was purified by column chromatography (C18 silanized silica gel (40-63 µ) eluting with methanol/water = 4:1) and triturated with diitopropyl ether, and N1-[(1S)-2,2-dimethyl- 1-([(1R)-1-(3-pyridyl)ethylamino]carbonyl)propyl]-(N4-hydroxy)-(2R)-2-{3-[3-methyl-(4-phenyl)phenyl]propyl }butanediamide (122 mg, 60%) as a white solid.

T.t. 112-114 °C.

Rf 0.33 (C18 silanized silica gel, methanol/water = 5:1).

δH (400 MHz, CH3OD) 1.01 (9H, s); 1.46 (3H, d, J = 7 Hz); 1.40-1.64 (4H, m); 2.16 (3H, s); 2.20 (1H, m); 2.36 (1H, m); 2.50 (2H, m); 2.88 (1H, m); 4.32 (1H, s); 5.02 (1H, q, J = 7 Hz); 6.92 (1H, m); 7.00 (2H, m); 7.18 (1H, m); 7.22 (2H, m); 7.32 (1H, m); 7.40 (2H, m); 7.72 (1H, m); 8.28 (1H, m); 8.48 (1H, s).

LRMS (thermospray) m/z = 559 (MH+).

Found: C, 70.13; H, 7.83; N 9.48.

C33H42N4O4 • 0.33 H2O • 0.33 DIPE requirements: C, 70.20; H, 7.80; N 9.63%.

Example 25

(N4,3S)-dihydroxy-N1-[(1S)-2,2-dimethyl-1-({[(1R)-1-(4-pyridyl)ethyl]amino}carbonyl)propyl]-(2R)- 2-{3-[3-methyl-(4-phenyl)phenyl]propyl}butanediamide

[image]

(R)-(+)-1-(4-pyridyl)ethylamine was prepared by the procedure according to "J. Amer. Chem. Soc.", 95, 811, (1973), by recrystallization of (-) tartaric acid salt of racemic amine, [α]D2O = -16.7 ° (c = 0.20, H2O). The free base was released by dissolving the tartrate salt in an excess of aqueous sodium hydroxide solution saturated with sodium chloride and extracted several times with dichloromethane. The free base was used directly without purification.

a) 7-azabenzotriazol-1-yloxytris(pyrrolidino)phosphonium hexafluorophosphate (208 mg, 0.40 mmol) was added to a stirred solution of (2R)-N-[(1S)-1-(carboxy)-2,2-dimethylpropyl] -2-[(4S)-2,2-dimethyl-5-oxo-1,3-dioxolan-4-yl]-5-[(3-methyl-4-phenyl)phenyl]pentanamide (preparation 33) (198 mg, 0.40 mmol), (R)-(+)-1-(4-pyridyl)ethylamine (49 mg, 0.40 mmol) and collidine (53 µl, 0.40 mmol) in anhydrous dichloromethane (4 ml ) under nitrogen at 0 °C. After 1 hour, the mixture was stirred at 20 °C for 3 hours. The mixture was then poured into ethyl acetate (100 ml) and water (100 ml), and sodium chloride was added to separate the phases. The layers were separated, and the organic layer was washed with a 5% aqueous solution of sodium bicarbonate (2 x 75 ml) with the addition of more solid sodium chloride. The organic solution was dried (Na2SO4) and concentrated under reduced pressure. The residue was purified by flash chromatography (gradient elution with hexane:ethyl acetate = 5:1 to 1:5) and a solid was obtained, which was triturated with diisopropyl ether, after which (2R)-2-[(4S)-2,2 was obtained -dimethyl-5-oxo-1,3-dioxolan-4-yl]-N-[(1S)-2,2-dimethyl-1-({[1R)-1-(4-pyridyl)ethyl]amino} carbonyl)propyl]-5-[(3-methyl-4-phenyl)phenyl]pentanamide (104 mg, 43%) as a white solid.

T.t. 167-170 °C.

Rf 0.10 (hexane:ethyl acetate = 1.3).

δH (400 MHz, DMSO-d6) 0.92 (9H, s); 1.34 (3H, d, J = 7 Hz); 1.45 (3H, s, and 2H, m, overlap); 1.56 (3H, s, and 1H, m, overlap); 1.72-1.82 (1H, m); 2.14 (3H, s); 2.40-2.60 (2H, m); 2.98 (1H, m); 4.40 (1H, d, J= 9 Hz); 4.46 (1H, d, J = 9 Hz); 4.90 (1H, pentet, J= 7 Hz); 6.94 (1H, m); 7.00 (2H, m); 7.20 (2H, d, J = 5.5 Hz); 7.30 (3H, m); 7.40 (2H, m); 7.86 (1H, d, J = 9 Hz); 8.30 (2H, d, J= 5.5 Hz); 8.58 (1H, d, J = 7 Hz).

LRMS (thermospray) m/z = 600 (MH+).

Found: C, 69.04; H 7.31; N 6.68.

C36H45N3O5 • 1.2 H2O • 0.13 EtOAc required: C, 69.32; H 7.71; N 6.64 %.

b) According to the procedure from example 19, (2R)-2-[(4S)-2,2-dimethyl-5-oxo-1,3-dioxolan-4-yl]-N-[(1S)-2,2 -dimethyl-1-({[(1R)-1-(4-pyridyl)ethyl]amino}carbonyl)propyl]-5-[(3-methyl-4-phenyl)phenyl]pentanamide (from a, above) ( 86 mg, 0.14 mmol) was reacted with hydroxylamine at room temperature for 18 hours. The solution was concentrated under reduced pressure and purified by column chromatography (C18 silanized silica gel (40-63 µ) eluting with methanol:water = 3:1). The residue was azeotroped with ethanol, then ethyl acetate and triturated with diisopropyl ether to give the title compound as a white solid (45 mg, 56 %).

T.t. 100-110 °C.

Rf 0.10 (dichloromethane:methanol:concentrated aqueous ammonia solution = 90:10:1).

δH (400 MHz, DMSO-d6) 0.92 (9H, s); 1.26 (1H, m, and 3H, d, J= 7 Hz, overlap); 1.36-1.60 (3H, m); 2.10 (3H, s); 2.40 (1H, m); 2.48 (1H, m); 2.70 (1H, m); 3.80 (1H, t, J = 7.5 Hz); 4.34 (1H, d, J = 9.5 Hz); 4.88 (1H, pentet, J = 7 Hz); 5.24 (1H, d, J = 7.5 Hz); 6.90 (1H, m); 6.98 (2H, m); 7.20 (2H, d, J = 5.5 Hz); 7.28 (3H, m); 7.40 (2H, m); 7.60 (1H, d, J = 9.5 Hz); 8.32 (2H, d, J = 5.5 Hz); 8.42 (1H, d, J = 7 Hz); 8.80 (1H, s); 10.60 (1H, no s).

Found: C, 67.88; H, 7.44; N, 9.42.

C33H42N4O5 • 0.5 H2O requirements: C, 67.90; H, 7.43; N, 9.06%.

Example 26

(N4,3S)-dihydroxy-N1-[(1S)-2,2-dimethyl-1-({[(1R)-1-(3-pyridyl)ethyl]amino}carbonyl)propyl]-(2R)- 2-{3-[3-methyl-(4-phenyl)phenyl]propyl}butanediamide

[image]

(R)-(+)-1-(3-pyridyl)ethylamine was prepared by the procedure of "J. Amer. Chem. Soc.", 95, 811, (1973), by recrystallization of the (-)-tartaric acid salt of the racemic amine , [α]D2O = -20.1 ° (c = 1.67, H2O). The free base was released by dissolving the tartrate in an excess of 1M aqueous sodium hydroxide solution saturated with sodium chloride and extracted several times with dichloromethane. The free base was used directly without purification.

a) According to the procedure from example 25(a), (2R)-N-[(1S)-1-(carboxy)-2,2-dimethyl]propyl]-2-[(4S)-2,2-dimethyl- 5-oxo-1,3-dioxolan-4-yl]-5-[(3-methyl-4-phenyl)phenyl]pentanamide (preparation 33) (86 mg, 0.14 mmol) was reacted with the free base (R)-(+)-1-(3-pyridyl)ethylamine (49 mg, 0.40 mmol) at room temperature for 3 hours, and then by the same treatment and trituration of the crude product with diisopropyl ether, (2R)-2-[ (4S)-2,2-dimethyl-5-oxo-1,3-dioxolan-4-yl]-N-[(1S)-2,2-dimethyl-1-({[(1R)-1-( 3-Pyridyl)ethyl]amino}carbonyl)propyl]-5-[(3-methyl-4-phenyl)phenyl]pentanamide (165 mg, 69%) as a white solid.

T.t. 172-176 °C.

Rf 0.10 (hexane:ethyl acetate = 1:3).

δH (400 MHz, DMSO-d6) 0.92 (9H, s); 1.35 (3H, d, J = 7.5 Hz); 1.45 (2H, m, and 3H, s, overlap); 1.55 (1H, m, and 3H, s, overlap); 1.75 (1H, m); 2.12 (3H, s); 2.38-2.58 (2H, m); 2.95 (1H, m); 4.35 (1H, d, J= 10 Hz); 4.45 (1H, d, J= 9.5 Hz); 4.98 (1H, pentet, J = 7.5 Hz); 6.94 (1H, m); 7.00 (2H, m); 7.10 (1H; m); 7.25 (2H, m); 7.35 (1H, m); 7.40 (2H, m); 7.68 (1H, d, J= 8 Hz); 7.82 (1H, d, J = 9.5 Hz); 8.32 (1H, m); 8.45 (1H, s); 8.55 (1H, d, J= 7.5 Hz).

LRMS (thermospray) m/z = 600 (MH+).

b) According to the procedure from example 19, (2R)-2-[(4S)-2,2-dimethyl-5-oxo-1,3-dioxolan-4-yl]-N-[(1S)-2,2 -dimethyl-1-({[(1R)-1-(3-pyridyl)ethyl]amino}carbonyl]propyl-5-[(3-methyl-4-phenyl)phenyl]pentathanamide (from a, above), ( 142 mg, 0.24 mmol) was reacted with hydroxylamine at room temperature for 17 hours. The solution was concentrated under reduced pressure and purified by column chromatography (C18 silanized silica gel (40-63 µ) with gradient elution with methanol:water = 60 :40 to 80:20), and a yellowish solid was obtained. It was purified again by reverse phase chromatography (eluting with methanol:water = 75:25). The product was then azeotroped with ethanol, ethyl acetate and triturated with diisopropyl ether, and obtained is the title compound as a white solid (50 mg, 36 %).

T.t. 85-95 °C.

Rf 0.11 (dichloromethane:methanol:concentrated aqueous ammonia solution = 90:10:1).

δH (400 MHz, DMSO-d6) 0.90 (9H, s); 1.25 (1H, m); 1.30 (3H, d, J= 7 Hz); 1.36-1.55 (3H, m); 2.15 (3H, s); 2.40 (2H, m); 2.70 (1H, m); 3.78 (1H, t, J = 7.5 Hz); 4.30 (1H, d, J= 9.5 Hz); 4.94 (1H, pentet, J= 7 Hz); 5.24 (1H, d, J = 7.5 Hz); 6.90 (1H, m); 7.00 (2H, m); 7.12 (1H, m); 7.20-7.38 (3H, m); 7.40 (2H, m); 7.60 (2H, m); 8.32 (1H, m); 8.38 (1H, d, J = 7 Hz); 8.50 (1H, s); 8.82 (1H, no s); 10.55 (1H, no s).

LRMS (thermospray) m/z = 514 (base peak, M+ - HONHCO).

Found: C, 67.52; H, 7.44; N, 9.52.

C33H42N4O5 • 0.67 H2O requirements: C, 67.57; H, 7.44; N, 9.55%.

Example 27

(N4,3S)-dihydroxy-N1-[(1S)-2,2-dimethyl-1-({[(1S)-2-hydroxy-1-phenylethyl]amino}carbonyl)propyl]-(2R)-2 -{3-[3-methyl-(4-phenyl)phenyl]propyl}butanediamide

[image]

a) According to the procedure from example 25(a), (2R)-N-[(1S)-1-(carboxy)-2,2-dimethylpropyl]-2-[(4S)-2,2-dimethyl-5- oxo-1,3-dioxolan-4-yl]-5-[(3-methyl-4-phenyl)phenyl]pentanamide (preparation 33) (150 mg, 0.30 mmol) was reacted with (S)- with (+)-2-phenylglycinol (41 mg, 0.30 mmol) at room temperature for 3 hours. The mixture was taken up in ethyl acetate (75 ml) and washed with 0.5 M aqueous dihydrogen phosphate solution (2 x 50 ml) (sodium chloride was added to cause phase separation). The organic solution was dried (Na2SO4) and concentrated under reduced pressure. The oily residue was triturated with diethyl ether to give (2R)-2-[(4S)-2,2-dimethyl-5-oxo-1,3-dioxolan-4-yl]-N-[(1S)-2, 2-dimethyl-1-({[(1S)-2-hydroxy-1-phenylethyl]amino}carbonyl)propyl]-5-[(3-methyl-4-phenyl)phenyl]pentanamide (80 mg, 43%) in the form of a white solid.

T.t. 206-209 °C

Rf 0.33 (hexane:ethyl acetate:acetic acid = 50:50:1).

δH (400 MHz, DMSO-d6) 0.95 (9H, s); 1.42 (2H, m, and 3H, s, overlap); 1.56 (1H, m, and 3H, s, overlap); 1.75 (1H, m); 2.16 (3H, s); 2.48 (2H, m); 2.95 (1H, m); 3.55 (2H, m); 4.40 (1H, d, J= 9.5 Hz); 4.48 (1H, d, J = 9.5 Hz); 4.75 (1H, t, J= 5.5 Hz); 4.84 (1H, q, J = 7 Hz); 6.94 (1H, m); 7.00 (2H, m); 7.10 (3H, m); 7.20 (2H, m); 7.30 (3H, m); 7.40 (2H, m); 7.80 (1H, d, J = 9.5 Hz); 8.32 (1H, d, J = 7 Hz).

LRMS (thermospray) m/z = 615 (MH+).

Found: C, 71.46; H, 7.53; N, 4.59.

C37H46N2O6 • 0.33 H2O requirements: C, 71.59; H, 7.58; N, 4.51%.

b) According to the procedure from example 19, (2R)-2-[(4S)-2,2-dimethyl-5-oxo-1,3-dioxolan-4-yl]-N-[(1S)-2,2 -dimethyl-1-({[(1S)-2-hydroxy-1-phenylethyl]amino}carbonyl)propyl]-5-[(3-methyl-4-phenyl)phenyl]pentanamide (from a, above) (66 mg, 0.11 mmol) was reacted with hydroxylamine at room temperature for 17 hours. The solution was concentrated under reduced pressure and purified by column chromatography (C18 silanized silica gel (40-63 µ) with gradient elution with methanol:water = 60:40 to 80:20). The residue was azeotroped with ethanol followed by ethyl acetate and triturated with diisopropyl to give the title compound as a yellowish solid (35 mg, 54 %).

T.t. 80-90 °C.

Rf 0.06 (dichloromethane:methanol:concentrated aqueous ammonia solution = 90:10:1).

δH (400 MHz, DMSO-d6) 0.92 (9H, s); 1.22 (1H, m); 1.30-1.55 (3H, m); 2.15 (3H, s); 2.40 (2H, m); 2.68 (1H, m); 3.56 (2H, m); 3.78 (1H, t, J = 7.5 Hz); 4.38 (1H, d, J = 9.5 Hz); 4.75 (1H, m); 4.82 (1H, q, J = 7.5 Hz); 5.22 (1H, d, J = 7.5 Hz); 6.90 (1H, m); 7.00 (2H, m); 7.15 (3H, m); 7.20-7.38 (5H, m); 7.40 (2H, m); 7.58 (1H, d, J = 9.5 Hz); 8.20 (1H, d, J = 7.5 Hz); 8.80 (1H, no s); 10.58 (1H, no s).

LRMS (thermospray) m/z = 529 (base peak, M+ - HONHCO).

Found: C, 67.91; H, 7.71; N, 6.58.

C34H43N3O6 • 0.67 H2O • 0.25 DIPE requirements: C, 67.99; H, 7.69; N, 6.70%.

Example 28

(N4,3S)-dihydroxy-N1-[(1S)-2,2-dimethyl-1-({[(1R)-2,3-dihydro-1H-inden-1-yl]amino}carbonyl)propyl] -(2R)-2-{3-[3-methyl-(4-phenyl)phenyl]propyl}butanediamide

[image]

a) According to the procedure from example 25(a), (2R)-N-[(1S)-1-(carboxy)-2,2-dimethylpropyl]-2-[(4S)-2,2-dimethyl-5- oxo-1,3-dioxolan-4-yl]-5-[(3-methyl-4-phenyl)phenyl]pentanamide (preparation 33) (150 mg, 0.30 mmol) was reacted with (R)- with (-)-1-aminoindane (40 mg, 0.30 mmol) at room temperature for 3 hours. The mixture was poured into ethyl acetate (75 ml) and washed with 0.5 M aqueous sodium dihydrogen phosphate (2 x 50 ml) and with 5% aqueous sodium bicarbonate (50 ml) (solid sodium chloride was added to separate the phases ). The organic solution was dried (Na2SO4) and concentrated under reduced pressure. The residue in the form of oil was purified by flash chromatography (gradient elution with hexane:ethyl acetate = 10:1 to 2:1), and then triturated with diisopropyl ether to obtain (2R)-2-[(4S)-2,2-dimethyl -5-oxo-1,3-dioxolan-4-yl]-N-[(1S)-2,2-dimethyl-1-({[(1R)-2,3-dihydro-1H-inden-1- yl]amino}carbonyl)propyl]-5-[(3-methyl-4-phenyl)phenyl]pentanamide, as a white solid (135 mg, 74 %).

Rf 0.76 (hexane: ethyl acetate: acetic acid = 50:50:1).

δH (400 MHz, CDCl 3 ) 1.02 (9H, s); 1.55 (3H, s); 1.60 (3H, s); 1.65-2.00 (5H, complex); 2.20 (3H, s); 2.60 (1H, m); 2.66 (3H, m); 2.86 (1H, m); 2.95 (1H, m); 4.20 (1H, d, J= 10 Hz); 4.54 (1H, d, J= 7 Hz); 5.42 (1H, q, J = 8 Hz); 5.82 (1H, d, J = 7 Hz); 6.62 (1H, d, J= 8 Hz); 7.00-7.40 (12H, complex).

Found: C, 74.06; H, 7.61; N, 4.47.

C38H46N2O5 • 0.33 EtOAc required: C, 73.81; H, 7.66; N, 4.38%.

b) According to the procedure from example 19, (2R)-2-[(4S)-2,2-dimethyl-5-oxo-1,3-dioxolan-4-yl]-N-[(1S)-2,2 -dimethyl-1-({[(1R)-2,3-dihydro-1H-inden-1-yl]amino}carbonyl)propyl]-5-[(3-methyl-4-phenyl)phenyl]pentanamide (from a, above) (105 mg, 0.17 mmol) was reacted with hydroxylamine at room temperature for 17 h. The solution was concentrated under reduced pressure and purified by column chromatography (C18 silanized silica gel (40-63 µ) with gradient elution with methanol:water = 60:40 to 85:15). The residue was azeotroped with ethanol, then ethyl acetate, then triturated with diisopropyl ether to give the title compound as a yellowish solid (60 mg, 60%).

T.t. 100-120 °C.

Rf 0.10 (dichloromethane:methanol:concentrated aqueous ammonia solution = 90:10:1).

δH (400 MHz, DMSO-d6) 0.92 (9H, s); 1.26 (1H, m); 1.42-1.60 (3H, m); 1.65 (1H, m); 2.15 (3H, s); 2.22 (1H, m); 2.40-2.62 (2H, m); 2.72 (2H, m); 2.80 (1H, m); 3.80 (1H, t, J= 8 Hz); 4.30 (1H, d, J= 9.5 Hz); 5.25 (2H, m); 6.90-7.45 (12H, complex); 7.62 (1H, d, J = 9.5 Hz); 8.20 (1H, d, J= 8.5 Hz); 8.80 (1H, s); 10.58 (1H, no s).

LRMS (thermospray) m/z = 526 (base peak, MH+ - HONHCO).

Found: C, 70.94; H, 7.50; N, 6.81.

C35H43N3O5 • 0.33 H2O requirements: C, 71.05; H, 7.44; N, 7.10%.

Example 29

(N4,3S)-dihydroxy-N1-[(1S)-2,2-dimethyl-1-({[(1S)-2-methoxy-1-phenylethyl]amino}carbonyl)propyl]-(2R)-2 -{3-[3-methyl-(4-phenyl)phenyl]propyl}butanediamide

[image]

a) According to the procedure from example 25(a), (2R)-N-[(1S)-1-(carboxy)-2,2-dimethylpropyl]-2-[(4S)-2,2-dimethyl-5- oxo-1,3-dioxolan-4-yl]-5-[(3-methyl-4-phenyl)phenyl]pentanamide (preparation 33) (198 mg, 0.40 mmol) was reacted with (1S)- with 2-methoxy-1-phenylethylamine (61 mg, 0.40 mmol) at room temperature for 3 hours. The mixture was then poured into ethyl acetate (75 ml) and washed with 0.5 M aqueous dihydrogen phosphate (2 x 50 ml) and 5% aqueous sodium bicarbonate (50 ml) (solid sodium chloride was added to separate the phases). The organic solution was dried (Na2SO4) and concentrated under reduced pressure. The residual oil was purified by flash chromatography (gradient elution with hexane:ethyl acetate = 4:1 to 1:1) and triturated with diisopropyl ether to give (2R)-2-[(4S)-2,2-dimethyl-5- oxo-1,3-dioxolan-4-yl]-N-[(1S)-2,2-dimethyl-1-({[(1S)-2-methoxy-1-phenylethyl]amino}carbonyl)propyl]- 5-[(3-methyl-4-phenyl)phenyl]pentanamide as a white solid (172 mg, 68 %).

Rf 0.53 (hexane:ethyl acetate:acetic acid = 50:50:1).

δH (400 MHz, CDCl 3 ) 1.02 (9H, s); 1.50 (3H, s); 1.55 (3H, s); 1.54 (2H, m); 1.90 (2H, m); 2.20 (3H, s); 2.60 (3H, m); 3.30 (3H, s); 3.60 (2H, d, J= 5 Hz); 4.28 (1H, d, J = 9 Hz); 4.50 (1H, d, J= 6Hz); 5.10 (1H, dt, J= 5 and 7 Hz); 6.36 (1H, d, J= 7Hz); 6.50 (1H, d, J= 9Hz); 6.98 (1H, d, J= 8 Hz); 7.01 (1H, s); 7.10 (1H, d, J= 8 Hz); 7.15-7.36 (8H, complex); 7.40 (2H, m).

LRMS (thermospray) m/z = 629 (MH+); 651 (MNa+).

Found: C, 72.41; H, 7.72; N, 4.48.

C38H48N2O6 required: C, 72.49; H, 7.72; N, 4.48%.

b) According to the procedure from example 9, (2R)-2-[(4S)-2,2-dimethyl-5-oxo-1,3-dioxolan-4-yl]-N-[(1S)-2,2 -dimethyl-1-({[(1S)-2-methoxy-1-phenylethyl]amino}carbonyl)propyl]-5-[(3-methyl-4-phenyl)phenyl]pentanamide (from a, above) (142 mg, 0.22 mmol) was reacted with hydroxylamine at room temperature for 17 hours. The solution was concentrated under reduced pressure and purified by column chromatography (C18 silanized silica gel (40-63 µ) eluting with methanol:viodom = 3:1). The residue was azeotroped with ethanol, then ethyl acetate and triturated with diisopropyl ether to give the title compound as a white solid (109 mg, 82 %).

T.t. 100-110 °C.

Rf 0.07 (dichloromethane:methanol:concentrated aqueous ammonia solution = 90:10:1).

δH (400 MHz, DMSO-d6) 0.90 (9H, s); 1.20 (1H, m); 1.30-1.50 (3H, m); 2.10 (3H, s); 2.38 (1H, m); 2.44 (1H, m); 2.65 (1H, m); 3.17 (3H, s); 3.45 (2H, d, J = 7.5 Hz); 3.75 (1H, t, J= 8 Hz); 4.40 (1H, d, J= 9.5 Hz); 5.00 (1H, q, J= 7.5 Hz); 5.22 (1H, d, J= 8 Hz); 6.90 (1H, d); 6.98 (2H, m); 7.15 (3H, m); 7.25 (4H, m); 7.35 (1H, m); 7.40 (2H, m); 7.58 (1H, d, J = 9.5 Hz); 8.35 (1H, d, J = 7.5 Hz); 8.80 (1H, no s); 10.58 (1H, no s).

LRMS (thermospray) m/z = 575 (MH+); 514 (M+ - HONHCO).

Found: C, 68.71; H, 7.64; N, 6.65.

C35H45N3O6 • 0.5 H2O requirements: C, 68.60; H, 7.57; N, 6.86%.

Example 30

(N4,3S)-dihydroxy-N1-[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]-(2R)-2-{3- [3'-Methoxy-2-methylbiphen-4-yl)propyl]}butanediamide

[image]

a) According to the procedure from example 25(a), (2R)-N-[(1S)-1-(carboxy)-2,2-dimethylpropyl]-2-[(4S)-2,2-dimethyl-5- oxo-1,3-dioxolan-4-yl]-5-[(3'-methoxy-2-methylbiphen-4-yl)propyl]pentanamide (preparation 34) (150 mg, 0.28 mmol) was added to the reaction with (R)-1-phenylethylamine (35 µl, 0.28 mmol) at room temperature for 2.5 hours. The mixture was taken up in ethyl acetate (75 ml) and washed with 0.5 M aqueous sodium dihydrogen phosphate (2 x 50 ml) and 5% aqueous sodium bicarbonate (50 ml) (sodium chloride was added to separate the phases). The organic solution was dried (Na2SO4) and concentrated under reduced pressure. The remaining oil was purified by flash chromatography (eluting with hexane:ethyl acetate = 35:65) and (2R)-2-[(4S)-2,2-dimethyl-5-oxo-1,3-dioxolan-4- yl]-N-[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]-5-[(3'-methoxy-2-methylbiphen-4 -yl)propyl]pentanamide as a white solid (133 mg, 76 %).

Rf 0.49 (hexane:ethyl acetate:acetic acid = 50:50:1).

δH (300 MHz, CDCl 3 ) 1.00 (9H, s); 1.40-1.60 (11H, complex); 1.62 (1H, m); 1.90 (1H, m); 2.21 (3H, s); 2.60 (3H, m); 3.80 (3H, s); 4.18 (1H, d, J= 9 Hz); 4.45 (1H, d, J= 7 Hz); 5.05 (1H, pentet, J= 7.5 Hz); 5.85 (1H, d, J = 7.5 Hz); 6.50 (1H, d, J= 9 Hz); 6.84 (3H, m); 7.00 (2H, m); 7.14 (1H, d, J = 9.5 Hz); 7.18-7.40 (6H, complex).

LRMS (thermospray) m/z = 646 (MNH4+).

b) According to the procedure from example 19, (2R)-2-[(4S)-2,2-dimethyl-5-oxo-1,3-dioxolan-4-yl]-N-[(1S)-2,2 -dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]-5-[(3'-methoxy-2-methylbiphen-4-yl)propyl]pentathanamide (from a, above) ( 133 mg, 0.21 mmol) was reacted with hydroxylamine (59 mg, 0.85 mmol) at room temperature for 18 hours. The solution was concentrated under reduced pressure, the residue was purified by column chromatography (C18 silanized silica gel (40-63 µ), with gradient elution with methanol:water = 3:2 to 4:1), then it was triturated with diisopropyl ether and the title compound was obtained as a white solid (57 mg, 45 %).

T.t. 98-100 °C.

Rf 0.20 (dichloromethane: methanol = 95:5).

δH (400 MHz, DMSO-d6) 0.92 (9H, s); 1.18-1.30 (1H, m); 1.25 (3H, d, J= 7.5 Hz); 1.38-1.58 (3H, m); 2.15 (3H, s); 2.40 (1H, m); 2.46 (1H, m); 2.70 (1H, m); 3.76 (4H, m); 4.35 (1H, d, J= 9.5 Hz); 4.90 (1H, pentet, J= 7.5 Hz); 5.22 (1H, d, J= 8 Hz); 6.75-7.40 (12H, complex); 7.58 (1H, d, J = 9.5 Hz); 8.26 (1H, d, J = 7.5 Hz); 8.80 (1H, s); 10.58 (1H, s).

LRMS (thermospray) m/z = 621 (MNH4+).

Found: C, 69.17; H, 7.55; N, 6.87.

C35H45N3O6 • 0.2 H2O requirements: C, 69.22; H, 7.53; N, 6.92%.

Example 31

(N4,3S)-dihydroxy-N1-[(1S)-2,2-dimethyl-1-({[(1S)-2-methoxy-1-phenylethyl]amino}carbonyl)propyl]-(2R)-2 -{3-[(3'-methoxy-2-methylbiphen-4-yl)propyl}butanediamide

[image]

a) According to the procedure from example 25(a), (2R)-N-[(1S)-1-(carboxy)-2,2-dimethylpropyl]-2-[(4S)-2,2-dimethyl-5- oxo-1,3-dioxolan-4-yl]-5-[(3'-methoxy-2-methylbiphen-4-yl)propyl]pentanamide (preparation 34) (220 mg, 0.42 mmol) was added to the reaction with (1S)-2-methoxy-1-phenylethylamine (63 mg, 0.42 mmol) at room temperature for 2.5 hours. The mixture was taken up in ethyl acetate (75 ml) and washed with 0.5 M aqueous dihydrogen phosphate (2 x 50 ml) and 5% aqueous sodium bicarbonate (50 ml) (solid sodium chloride was added to separate the phases). The organic solution was dried (Na2SO4) and concentrated under reduced pressure. The residue was purified by flash chromatography (gradient elution with hexane:ethyl acetate = 30:70 to 60:40) and a white solid was obtained, which was purified again by flash chromatography (sorbsil C60, (20-40 µ) silica gel, eluting with hexane) :ethyl acetate = 35:65) and obtained (2R)-2-[(4S)-2,2-dimethyl-5-oxo-1,3-dioxolan-4-yl]-N-[(1S)- 2,2-dimethyl-1-({[(1S)-2-methoxy-1-phenylethyl]amino}carbonyl)propyl]-5-[(3'-methoxy-2-methylbiphen-4-yl)propyl]pentanamide as a white solid (191 mg, 69 %).

Rf 0.37 (hexane:ethyl acetate = 1:1).

δH (400 MHz, CDCl 3 ) 1.00 (9H, s); 1.50 (3H, s); 1.55 (3H, s); 1.65 (2H, m); 1.86 (2H, m); 2.20 (3H, s); 2.60 (3H, m); 3.35 (3H, s); 3.60 (2H, d, J= 5 Hz); 3.80 (3H, s); 4.25 (1H, d, J= 9 Hz); 4.45 (1H, d, J= 6 Hz); 5.10 (1H, dt, J= 5 and 7.5 Hz); 6.35 (1H, d, J = 7.5 Hz); 6.50 (1H, d, J= 9 Hz); 6.80-6.92 (3H, m); 6.96-7.04 (2H, m); 7.10 (1H, d, J= 8 Hz); 7.15-7.35 (6H, complex).

LRMS (thermospray) m/z = 659 (MH+).

b) According to the procedure from example 19, (2R)-[(4S)-2,2-dimethyl-5-oxo-1,3-dioxolan-4-yl]-N-[(1S)-2,2-dimethyl -1-({[(1S)-2-methoxy-1-phenylethyl]amino}carbonyl)propyl]-5-[(3'-methoxy-2-methylbiphen-4-yl)propyl]pentanamide (from a, above ) (191 mg, 0.29 mmol) was reacted with hydroxylamine at room temperature for 17 hours. The solution was concentrated under reduced pressure, and the residue was purified by column chromatography (C18 silanized silica gel (40-60 µ), with gradient elution with methanol:water = 60:40 to 70:30) and trituration with diisopropyl ether, and the compound from title as a white solid (81 mg, 44 %).

T.t. 82-89 °C.

Rf 0.40 (dichloromethane:methanol = 90:10).

δH (400 MHz, DMSO-d6) 0.90 (9H, s); 1.20 (1H, m); 1.40 (3H, m); 2.15 (3H, s); 2.38 (1H, m); 2.45 (1H, m); 2.70 (1H, m); 3.20 (3H, s); 3.44 (2H, d, J = 7 Hz); 3.76 (4H, m); 4.38 (1H, d, J = 9.5 Hz); 5.00 (1H, q, J= 7 Hz); 5.21 (1H, d, J = 7.5 Hz); 6.78 (1H, s); 6.80 (1H, d, J= 8 Hz); 6.88 (2H, m); 7.00 (2H, m); 7.14 (3H, m); 7.24 (2H, m); 7.32 (1H, t); 7.58 (1H, d, J = 9.5 Hz); 8.32 (1H, d, J = 7 Hz); 8.80 (1H, no s); 10.58 (1H, no s).

LRMS (thermospray) m/z = 634 (MH+).

Found: C, 67.47; H, 7.52; N, 6.55.

C36H47N3O7 • 0.4 H2O requirements: C, 67.46; H, 7.52; N, 6.56%.

Example 32

(2R)-N1-[(1S)-2,2-dimethyl-1-({[(1S)-2-methoxy-1-phenylethyl]amino}carbonyl)propyl]-(N4-hydroxy)-2-{ 3-[3'-Methoxy-2-methylbiphen-4-yl]propyl}butanediamide

[image]

a) According to the procedure from example 2, (3R)-3-({[(1S)-2,2-dimethyl-1-({[(1S)-2-methoxy-phenylethyl]amino}carbonyl)propyl]amino} carbonyl)-6-[3'-methoxy-2-methylbiphen-4-yl]hexanoic acid (Example 15) (603 mg, 1.0 mmol) was reacted with O-allylhydroxylamine hydrochloride (134 mg, 1.2 mmol). Purification of the crude product by flash chromatography (with gradient elution with hexane:ethyl acetate = 1:1 to 1:2), and then by repeated flash chromatography (with dichloromethane:methanol:concentrated aqueous ammonia solution = 95:5:0.5) was obtained (2R)-N1-[(1S)-2,2-dimethyl-1-({[(1S)-2-methoxy-1-phenylethyl]amino}carbonyl)propyl]-2-{3-[3'- methoxy-2-methylbiphen-4-yl]propyl}-(N4-3-propenyloxy)butanediamide (397 mg, 60%) as a colorless foam.

T.t. 76-79 °C.

Rf 0.35 (hexane:ethyl acetate = 1:2).

δH (300 MHz, DMSO-d6) 0.92 (9H, s); 1.42 (4H, m); 2.01 (1H, dd, J= 7 and 15 Hz); 2.15 (3H, s, and 1H, m, overlap); 2.44 (1H, m); 2.55 (1H, m); 2.84 (1H, m); 3.20 (3H, s); 3.47 (2H, d, J = 8 Hz); 3.77 (3H, s); 4.20 (2H, m); 4.35 (1H, d, J= 10 Hz); 5.02 (1H, q, J = 7 Hz); 5.19 (1H, d, J= 10 Hz); 5.40 (1H, d, J= 17 Hz); 5.87 (1H, m); 6.78 (1H, s); 6.84 (1H, d, J= 8 Hz); 6.90 (2H, m); 7.00 (2H, m); 7.23 (6H, m); 7.70 (1H, no d, J= 10); 8.32 (1H, no d, J= 7 Hz); 10.85 (1H, no s).

LRMS (thermospray) m/z = 658 (base peak, MH+); 680 (MNa+).

νmax (KBr disk) 3310; 2970; 2930; 1640; 1537; 1481; 700 cm-1.

Found: C, 70.89; H, 7.85; N, 6.27.

C39H51N3O6 required: C, 71.21; H, 7.81; N, 6.39%.

b) According to the procedure from example 2, (2R)-N1-[(1S)-2,2-dimethyl-1-({[(1S)-2-methoxy-1-phenylethyl]amino}carbonyl)propyl]-2 -{3-[3'-Methoxy-2-methylbiphen-4-yl]propyl}-(N4-3-propenyloxy)butanediamide (380 mg, 0.58 mmol) was reacted with ammonium formate (366 mg, 5 .80 mmol) in ethanol/water (4:1, 10 ml) with palladium catalysis at reflux for 1 hour. After treatment, the residue was purified by reverse phase flash chromatography (C18 silanized silica gel 40-63 µ, eluting with methanol:water = 4:1), and (2R)-N1-[(1S)-2,2-dimethyl was obtained -1-({[(1S)-2-methoxy-1-phenylethyl]amino}carbonyl)propyl]-(N4-hydroxy)-2-{3-[3'-methoxy-2-methylbiphen-4-yl] propyl)butanediamide (260 mg, 73%) as a colorless solid.

T.t. 166-168 °C.

Rf 0.29 (reverse phase TLC, methanol:water = 4:1).

δH (300 MHz, methanol-d6) 1.03 (9H, s); 1.51 (4H, m); 2.16 (3H, s, and 1H, m, overlap); 2.35 (1H, dd, J= 8 and 13 Hz); 2.48 (2H, m); 2.87 (1H, m); 3.33 (3H, s); 3.58 (2H, d, J = 7 Hz); 4.40 (1H, s, ); 5.10 (1H, t, J= 7 Hz); 6.92 (1H, s); 6.80 (1H, d, J= 8 Hz); 6.89 (2H, no d, J= 8 Hz); 6.97 (1H, s); 6.98 (1H, d, J= 8 Hz); 7.15 (3H, m); 7.27 (3H, m).

νmax (KBr disk) 3290; 3240; 2960; 2930; 1644; 1531; 1481; 1226; 703 cm-1.

Found: C, 69.06; H, 7.80; N, 6.65.

C38H47N3O6 • 0.5 H2O requirements: C, 68.99; H, 7.72; N, 6.70%.

Preparation 1

(2S)-amino-3,3-dimethyl-N-[(1R)-1-phenylethyl]butanamide, hydrochloride

[image]

a) N-(dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (35.47 g, 185 mmol) was added to a stirred mixture of tert-butyl N-[(1S)-2,2-dimethyl-1-carboxy)propyl]carbamate ( 34.7 g, 150 mmol) (Fluka Chemicals; J. Pospisek, K. Blaha: "Coll. Czech. Chem. Commun.", 42, 1069-76, (1977)) and 1-hydroxy-1,2 ,3-benzotriazole hydrate (22.3 g, 165 mmol) in anhydrous dichloromethane (350 ml) under nitrogen at 4 °C. After 1 hour, (R)-(1-phenyl)ethylamine (19.4 g, 158 mmol) was added followed by N-methylmorpholine (16.7 g, 165 mmol). After 30 minutes, the mixture was allowed to warm to room temperature. After 17 hours at that temperature, the mixture was concentrated under reduced pressure and partitioned between ethyl acetate (400 ml) and water (400 ml). The organic layer was washed successively with 5% aqueous citric acid solution (2 x 400 ml) and saturated aqueous sodium bicarbonate solution (500 ml), dried (MgSO4) and concentrated under reduced pressure. The residue was triturated with diisopropyl ether, filtered and dried, after which (2S)-tert-(butoxycarbonyl)amino-3,3-dimethyl-N-[(1R)-1-phenylethyl]butanamide (47.5 g, 94 %) in the form of a colorless solid.

T.t. 166-169 °C.

Rf 0.7 (hexane:ether:acetic acid = 30:70:1).

δH (400 MHz, CDCl 3 ) 0.97 (9H, s); 1.39 (9H, s); 1.45 (3H, d, J= 7 Hz); 3.73 (1H, no d); 5.04 (1H, pentet, J= 7 Hz); 5.15 (1H, no s); 5.84 (1H, no d); 7.26 (5H, complex).

LRMS (thermospray) m/z = 330 (MH+).

FTIR νmax (KBr disc) 3340; 3280; 2980; 1713; 1696; 1642; 1179; 700 cm-1.

Found: C, 68.30; H, 9.08; N, 8.39.

C19H30N2O3 required: C, 68.23; H, 9.04; N, 8.38%.

b) (2S)-tert-(butoxycarbonyl)amino-3,3-dimethyl-N-[(1R)-1-phenylethyl]butanamide (46.4 g, 139 mmol) was dissolved in a mixture of anhydrous dichloromethane (600 ml) and dioxane (150 ml) and cooled to 4 °C. Hydrogen chloride was bubbled through the mixture with stirring until the solution became saturated. After stirring for 4 hours at 4 °C, the solution was concentrated under reduced pressure. The residue was triturated with ether and filtered to give the title compound (38.0 g, 100 %).

Rf 0.46 (hexane: isorpopanol: concentrated aqueous ammonia solution = 80:20:1).

δH (400 MHz, d6-DMSO) 1.03 (9H, s); 1.37 (3H, d, J = 7 Hz); 3.50 (1H, d, J= 8 Hz); 4.94 (1H, pentet, J= 7 Hz); 7.24 (1H, t, J= 8 Hz); 7.33 (2H, t, J = 8 Hz); 7.39 (2H, d, J = 8 Hz); 8.10 (3H, no s); 8.88 (1H, no. d).

LRMS (thermospray) m/z = 235 (MH+).

FTIR νmax (KBr disc) 2960; 1674; 1557; 1505; 700 cm-1.

Found: C, 61.98; H, 8.71; N, 10.09.

C14H22N2O • HCl • H2O requirements: C, 62.09; H, 8.56; N, 10.34%.

Preparation 2

tert-butyl (3R)-3-({[(1S)-2,2-dimethyl-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]amino}carbonyl)hex-5-enoate

[image]

N-(dimethylaminopropyl)-N'-ethylcarbodiimide (4.21 g, 22.0 mmol) was added to a stirred mixture of (2R)-2-[2-(tert-butoxy)-2-oxoethyl]pent-4-enoic acid (A.L. Castelhano, S.L. Bender, J.G. Deal, S. Horner, T.J. Liak, Z. Yuan: International Patent WO96/16027, (1996)) (3.80 g, 17.8 mmol) and 1-hydroxy-7- of azabenzotriazole (2.49 g, 18.3 mmol) in anhydrous dimethylformamide (60 mL) under nitrogen at 4 °C. After 1 hour (2S)-amino-3,3-dimethyl-N-[(1R)-1-phenylethyl]butanamide hydrochloride (preparation 1) (5.10 g, 18.8 mmol) was added, followed by diisopropylethylamine ( 2.42 g, 19.2 mmol). After another 30 minutes, the mixture was allowed to warm to room temperature. After 17 hours at that temperature, the mixture was concentrated under reduced pressure and partitioned between ethyl acetate (400 ml) and water (400 ml). The aqueous layer was saturated with sodium bicarbonate, then extracted with ethyl acetate (2 x 100 ml). The combined organic solutions were concentrated under reduced pressure, the residue was dissolved in ether (500 ml), washed with water (3 x 200 ml), dried (MgSO4) and concentrated under reduced pressure. The residue was triturated with pentane, filtered and dried, after which tert-butyl (3R)-3-({[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}) was obtained carbonyl)propyl]amino}carbonyl)hex-5-enoate (6.70 g, 88%), as a colorless solid.

T.t. 123-126 °C.

Rf 0.16 (hexane:isopropanol = 98:2).

δH (400 MHz, CDCl 3 ) 0.97 (9H, s); 1.39 (9H, s); 1.46 (3H, d, J = 7 Hz); 2.06 (1H, dt, 13 and 6.5 Hz); 2.09 (1H, m); 2.31 (1H, dd, J= 15 and 3 Hz); 2.71 (1H, m); 2.77 (1H, m); 4.14 (1H, d, J= 10 Hz); 4.90 (1H, d, J= 10 Hz); 4.94 (1H, d, J= 15 Hz); 5.05 (1H, pentet, J= 7 Hz); 5.42 (1H, m); 5.95 (1H, no d); 6.36 (1H, no d); 7.26 (5H, complex).

LRMS (thermospray) m/z = 431 (MH+).

FTIR νmax (KBr disk) 2635; 2098; 1733; 1640; 1540; 1363; 1153; 696 cm-1.

Found: C, 69.30; H, 8.96; N, 6.54.

C25H38N2O4 required: C, 69.74; H, 8.90; N, 6.51%.

Preparation 3

tert-butyl (3R)-3-({[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]amino}carbonyl)-6-[3 -methyl-(4-phenyl)phenyl]-hexanoate

[image]

a) A mixture of palladium acetate (52 mg, 0.23 mmol) and tri-(2-methylphenyl)phosphine (141 mg, 0.46 mmol) in anhydrous acetonitrile (10 ml) was sonicated at room temperature in for 1 minute until a cream colored suspension was formed. This suspension was added with a Pasteur pipette to the mixed solution of tert-butyl (3R)-3-({[(1S)-2,2-dimethyl-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]amino }carbonyl)hex-5-enoate (preparation 2) (861 mg, 2.0 mmol), 3-methyl-4-phenylbromobenzene (M. Gomberg, J.C. Pernet: "J. Amer. Chem. Soc.", 48, 1372-84, (1926), and also see preparation 19B) (1.32 g, 5.34 mmol) and triethylamine (1.28 ml, 9.29 mmol) in anhydrous acetonitrile (15 ml) under nitrogen. The mixture was sparged with nitrogen and then heated at reflux for 24 hours. The mixture was then poured into ethyl acetate (200 ml) and washed with saturated aqueous sodium chloride solution (2 x 100 ml), dried (Na 2 SO 4 ) and concentrated under reduced pressure. The residue was purified by flash chromatography (gradient elution with dichloromethane:ethyl acetate) and tert-butyl (3R,5E)-3-({[(1S)-2,2-dimethyl-1-({[(1R)- 1-Phenylethyl]amino}carbonyl)propyl]amino}carbonyl)-6-[3-methyl-(4-phenyl)phenyl]hex-5-enoate as a colorless foam (1.91 g, 69 %). 1H NMR suggested that two alkene isomers, (5Z) and (4E), were also present. The alkene mixture was transferred to the second step (see b, below).

Rf 0.52 (dichloromethane: ethyl acetate = 9:1).

δH (400 MHz, CDCl3) (for (5E) isomer), 1.00 (9H, s); 1.39 (9H, s); 1.47 (3H, d, J= 7 Hz); 2.26 (1H, m); 2.40 (2H, m); 2.60 (1H, dd, J= 9 and 15 Hz); 2.71 (1H, m); 4.20 (1H, d, J= 9 Hz); 5.02 (1H, pentet, J= 7 Hz); 6.04 (2H, m); 6.35 (1H, d, J= 14 Hz); 6.51 (1H, no d); 7.19 (11H, complex); 7.36 (2H, t, J = 8 Hz).

b) Solution of tert-butyl (3R,5E)-3-({[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]amino}carbonyl) -6-[3-methyl-(4-phenyl)phenyl]hex-5-enoate (1.91 g, 3.2 mmol) in ethanol (80 ml) was hydrogenated over 10% palladium on carbon (120 mg ) at 3 bar at 20 °C for 16 hours. The mixture was filtered through Arbocel filter aid and concentrated under reduced pressure to give (3R)-3-({[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino} carbonyl)propyl]amino}carbonyl)-6-[3-methyl-(4-phenyl)phenyl]hexanoate (2.07 g, 108 %) as a colorless foam, which was carried to the next step without purification (see example 1 ).

Rf 0.52 (dichloromethane: ethyl acetate = 9:1).

δH (400 MHz, CDCl 3 ) 0.99 (9H, s); 1.38 (9H, s); 1.47 (3H, d, J= 7 Hz); 1.52 (4H, complex); 2.20 (3H, s); 2.29 (1H, m); 2.53 (4H, m); 4.16 (1H, d, J= 9 Hz); 5.05 (1H, pentet, J= 7 Hz); 5.97 (1H, no d); 6.42 (1H, no d); 6.93 (1H, d, J= 8 Hz); 6.98 (1H, s); 7.07 (1H, d, J= 8 Hz); 7.23 (8H, complex); 7.36 (2H, t, J = 7 Hz).

LRMS (thermospray) m/z = 599 (MH+).

Found: C, 75.86; H, 8.44; N, 4.60.

C38H50N2O4 required: C, 76.22; H, 8.42; N, 4.68%.

Example 33 / Preparation 4

(3R)-6-[(3-chloro-4-phenyl)phenyl]-3-({[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl) )propyl]amino}carbonyl)hexanoic acid

[image]

a) A mixture of palladium acetate (24 mg, 0.1 mmol) and tri-(2-methylphenyl)phosphine (61 mg, 0.2 mmol) in anhydrous acetonitrile (1 ml) was sonicated at room temperature for 1 minute, while no cream-colored suspension was formed. This suspension was added using a Pasteur pipette to a stirred solution of tert-butyl (3R)-3-({[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl) propyl]amino}carbonyl}hex-5-enoate (preparation 2) (861 mg, 2.0 mmol), (3-chloro-4-phenyl)phenyl trifluoromethanesulfonate (preparation 9) (740 mg, 2.2 mmol) and of triethylamine (420 µl, 3.0 mmol) in anhydrous acetonitrile (3 ml) under nitrogen. The mixture was flushed with nitrogen, and then heated at reflux for 22 h. Further amounts of reagents and catalysts were added as follows: (3-chloro-4 -phenyl)phenyl trifluoromethanesulfonate (200 mg, 0.59 mmol), triethylamine (420 µl, 3.0 mmol), palladium acetate (35 mg, 0.145 mmol) and tri-(2-methylphenyl)phosphine (89 mg, 0, 29 mmol), and heating was continued for another 8 hours. The mixture was poured into ethyl acetate (100 ml) and washed with saturated aqueous sodium chloride solution (2 × 50 ml), dried (MgSO4) and concentrated under reduced pressure. The residue was flash purified by chromatography (with elution with toluene:ethyl acetate = 9:1) and obtained mostly ter c-butyl (3R,5E)-6-[3-chloro-(4-phenyl)phenyl]-3-({[(1S)-2,2-dimethyl-1-({[(1R)-1- phenylethyl]amino}carbonyl)propyl]amino}carbonyl)hex-5-enoate as a colorless foam (936 mg, 76 %). 1H NMR suggested that two alkene isomers, (5Z) and (4E), were also present. The alkene mixture was carried to the next step (see b, below).

Rf 0.14 (toluene:ethyl acetate = 10:1).

δH (400 MHz, CDCl 3 ) (for (5E) isomer), 0.96 (9H, s); 1.37 (9H, s); 1.45 (3H, d, J= 6.5 Hz); 2.29 (1H, m); 2.36 (1H, dd, J= 3 and 15 Hz); 2.45 (1H, m); 2.61 (1H, dd, J= 9 and 15 Hz); 2.73 (1H, m); 4.16 (1H, d, J = 9.5 Hz); 5.02 (1H, pentet, J= 6.5 Hz); 5.85 (1H, no d); 6.09 (1H, dt, J= 6.5 and 15 Hz); 6.32 (1H, d, J= 15 Hz); 6.49 (1H, no d); 7.18 (8H, complex); 7.37 (5H, complex).

LRMS (thermospray) m/z = 617 (MH+).

b) Mixture of tert-butyl (3R,5E)-6-[3-chloro-(4-phenyl)phenyl]-3-({[(1S)-2,2-dimethyl-1-({[(1R) -1-phenylethyl]amino}carbonyl)propyl]amino}carbonyl)hex-5-enoate (936 mg, 1.52 mmol) and p-toluenesulfonyl hydrazine (1.41 mg, 7.60 mmol) in toluene (15 mL ) was stirred under nitrogen at reflux for 4 hours. After cooling, the mixture was diluted with ether (150 ml) and washed with water (50 ml), 0.5 M hydrochloric acid (50 ml) and saturated aqueous sodium chloride solution (50 ml), dried (MgSO 4 ) and concentrated under reduced pressure. The residue was purified by flash chromatography (gradient elution with hexane/ethyl acetate) and tert-butyl (3R)-6-[3-chloro-(4-phenyl)phenyl]-3-({[(1S)-2, 2-Dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]amino}carbonyl)hexanoate (701 mg, 74%) as a colorless foam.

Rf 0.36 (hexane:ethyl acetate = 5:1).

δH (400 MHz, CDCl 3 ) 1.03 (9H, s); 1.43 (9H, s); 1.52 (3H, d, J= 7 Hz); 1.43-1.70 (4H, complex); 2.32 (1H, m); 2.60 (4H, m); 4.18 (1H, d, J= 9 Hz); 5.09 (1H, pentet, J = 7 Hz); 5.95 (1H, no d); 6.47 (1H, no d); 7.03 (1H, d, J= 8 Hz); 7.23 (7H, complex); 7.42 (5H, complex).

LRMS (thermospray) m/z = 620 (MH+).

c) tert-butyl (3R)-6-[3-chloro-(4-phenyl)phenyl]-3-({[(1S)-2,2-dimethyl-1-({[(1R)-1- Phenylethyl]amino}carbonyl)propyl]amino}carbonyl)hexanoate (665 mg, 1.07 mmol) was dissolved in anhydrous dioxane (30 mL) and cooled in an ice bath under nitrogen. Hydrogen chloride gas was bubbled through the stirred solution for 15 minutes, until it became saturated. The solution was stirred for 3 hours and then concentrated under reduced pressure. The residue was dissolved in toluene and concentrated under reduced pressure three times, and a colorless resin was obtained, which crystallized after addition of hexane and ethyl acetate. The white solid was recrystallized from ethyl acetate/diisopropyl ether to give the title compound (325 mg, 54 %).

T.t. 154-155.5 °C (from ethyl acetate/diisopropyl ether).

Rf 0.38 (ether:hexane:acetic acid = 70:30:1).

δH (400 MHz, CD3OD) (NH signals of only partially substituted ofamide were recorded) 1.03 (9H; s); 1.43 (3H, d, J = 6.5 Hz); 1.48 (2H, m, ); 1.52 (2H, m); 2.39 (1H, dd, J= 3 and 14 Hz); 2.53 (2H, m); 2.61 (1H, dd, J= 9 and 14 Hz); 2.86 (1H, m); 4.36 (1H, s); 5.00 (1H, pentet, J= 6.5 Hz); 7.03 (1H, d, J= 9 Hz); 7.17 (7H, complex); 7.36 (5H, complex); 7.76 (0.5H, no d); 8.47 (0.8H, no d).

LRMS (thermospray) m/z = 562 (MH+).

Found: C, 70.29; H, 6.98; N, 4.91.

C23H39ClN2O4 required: C, 70.38; H, 6.98; N, 4.97%.

Example 34 / Preparation 5

(3R)-3-({[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]amino}carbonyl)-6-(3-fluoro- 4-phenoxyphenyl)hexanoic acid

[image]

a) A mixture of palladium acetate (20 mg, 0.08 mmol) and tri-(2-methylphenyl)phosphine (50 mg, 0.16 mmol) was added to a stirred solution of tert-butyl (3R)-3-({[(1S )-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]amino}carbonyl)hex-5-enoate (preparation 2) (700 mg, 1.63 mmol), 3-fluoro-4-phenoxy-bromobenzene (preparation 10) (479 mg, 1.79 mmol) and triethylamine (340 µl, 2.43 mmol) in anhydrous acetonitrile (3 ml) under nitrogen. The mixture was flushed with nitrogen and then heated at reflux for 14 hours. Additional palladium acetate (20 mg, 0.08 mmol) and tri-(2-methylphenyl)phosphine (50 mg, 0.16 mmol) were added, and the mixture was heated for another 9 hours. After cooling, the mixture was poured into ethyl acetate (100 ml) and washed with saturated aqueous sodium chloride solution (2 x 50 ml), dried (MgSO 4 ) and concentrated under reduced pressure. The residue was purified by flash chromatography (eluting with hexane:ethyl acetate = 9:1) and mainly tert-butyl (3R,5E)-3-({[(1S)-2,2-dimethyl-1-({[ (1R)-1-phenylethyl]amino}carbonyl)propyl]amino}carbonyl)-6-(3-fluoro-4-phenoxyphenyl)hex-5-enoate as a colorless oil (949 mg). 1H NMR suggests that two alkene isomers, (5Z) and (4E), were present along with alkene starting material (10%). The alkene mixture was carried to the next step (see b, below).

Rf 0.24 (hexane:ethyl acetate = 3:1).

δH (300 MHz, CDCl 3 ) (for (5E) isomer) 1.02 (9H, s); 1.40 (9H, s); 1.50 (3H, d, J= 6 Hz); 2.26 (1H, m); 2.40 (2H, m); 2.64 (1H, dd, J= 9 and 17 Hz); 2.74 (1H, m); 4.23 (1H, d, J= 10 Hz); 5.06 (1H, pentet, J= 6 Hz); 5.98 (1H, m); 6.02 (1H, no d); 6.30 (1H, d, J= 15 Hz); 6.50 (1H, no d); 6.94 (3H, m); 7.07 (2H, complex); 7.27 (8H, complex).

LRMS (thermospray) m/z = 617 (MH+).

b) Solution of tert-butyl (3R)-3-({[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]amino}carbonyl)-6 -(3-fluoro-4-phenoxyphenyl)hexanoate (949 mg, 1.53 mmol), ammonium formate (474 mg, 7.51 mmol) and 10% palladium on carbon (100 mg) in methanol (10 ml) it was stirred at 20 °C for 16 hours. The mixture was filtered through an Arbocel filter aid and concentrated under reduced pressure. The residue was dissolved in ethyl acetate and washed with saturated aqueous sodium chloride, dried (MgSO4) and concentrated under reduced pressure to give tert-butyl (3R)-3-({[(1S)-2,2-dimethyl-1- ({[(1R)-1-phenylethyl]amino}carbonyl)propyl]amino}carbonyl)-6-(3-fluoro-4-phenoxyphenyl)hexanoate (876 mg, 92%), as a colorless foam, which was transferred in the next step without purification.

Rf 0.29 (hexane:ethyl acetate = 3:1).

δH (400 MHz, CDCl 3 ) 0.98 (9H, s); 1.26 (1H, m); 1.37 (9H, s); 1.44 (3H, m); 1.46 (3H, d, J = 6.5 Hz); 2.23 (1H, m); 2.40 (2H, m); 2.52 (2H, m); 4.32 (1H, d, J= 10 Hz); 5.08 (1H, pentet, J= 6.5 Hz); 6.48 (1H, no d); 6:58 (1H, no. d); 6.74 (1H, d, J= 8 Hz); 6.87 (3H, m); 7.01 (1H, t, J= 7 Hz); 7.19 (8H, complex).

LRMS (thermospray) m/z = 619 (MH+).

c) Trifluoroacetic acid (2 ml) was added dropwise over 5 minutes to a stirred solution of tert-butyl (3R)-3-({[(1S)-2,2-dimethyl-1-({[(1R) -1-phenylethyl]amino}carbonyl)propyl]amino}carbonyl)-6-(3-fluoro-4-phenoxyphenyl)hexanoate (876 mg, 1.14 mmol) in anhydrous dichloromethane (6 mL) under nitrogen at 20 °C . The solution was stirred for 16 hours and concentrated under reduced pressure. The residue was dissolved in toluene and concentrated under reduced pressure. (2x). Purification by flash chromatography (eluting with hexane:ethyl acetate:acetic acid = 75:25:1) afforded the title compound as a yellow resin (784 mg, 98 %).

Rf 0.21 (hexane:ethyl acetate:acetic acid = 75:25:1).

δH (400 MHz, CDCl 3 ) 0.96 (9H, s); 1.40 (3H, m); 1.47 (3H, d, J= 7 Hz); 1.58 (1H, m, ); 2.42 (3H, m); 2.68 (2H, m); 4.32 (1H, d, J=10); 5.04 (1H, pentet, J= 7 Hz); 6.35 (1H, no d); 6.73 (1H, d, J= 6 Hz); 6.82 (2H, m); 6.89 (2H, d, J = 8 Hz); 7.01 (1H, t, J= 6 Hz); 7.11 (1H, d, J= 8 Hz); 7.20 (7H, complex).

LRMS (APCI) m/z = 563 (MH + ).

Example 35 / Preparation 6

(3R)-3-({[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]amino}carbonyl)-6-(3-methyl- 4-phenoxyphenyl] hexanoic acid

[image]

a) A mixture of palladium acetate (20 mg, 0.08 mmol) and tri-(2-methylphenyl)phosphine (50 mg, 0.16 mmol) was added to a stirred solution of tert-butyl (3R)-3-({[(1S )-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]amino}carbonyl)hex-5-enoate (preparation 2) (578 mg, 1.23 mmol), 1-iodo-3-methyl-4-phenoxybenzene (preparation 11) (384 mg, 1.35 mmol), and triethylamine (300 µl, 2.14 mmol) in anhydrous acetonitrile (3 ml) under nitrogen. The mixture was flushed with nitrogen and then heated at reflux for 24 hours. After cooling, the mixture was poured into ethyl acetate (100 mL) and washed with saturated aqueous sodium chloride solution (2 x 50 mL), dried (MgSO 4 ), and concentrated under reduced pressure. The residue was purified by flash chromatography (eluting with hexane:ethyl acetate = 9:1) to 4:1) and mainly tert-butyl (3R, 5E)-3-({[(1S)-2,2-dimethyl- 1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]amino}carbonyl)-6-(3-methyl-4-phenoxyphenyl)hex-5-enoate as an orange foam (615 mg). 1H NMR suggests that two alkene isomers, (5Z) and (4E), were present along with the starting alkene material (10%). The alkene mixture was carried to the next step (see c, below).

Rf 0.5 (hexane:ethyl acetate = 3:1).

δH (300 MHz, CDCl 3 ) (for (5E) isomer) 1.02 (9H, s); 1.26 (9H, s); 1.50 (3H, d, J= 7 Hz); 2.19 (3H, s); 2.29 (1H, m); 2.44 (2H, m); 2.63 (1H, m); 2.74 (1H, m); 4.19 (1H, d, J= 10 Hz); 5.04 (1H, pentet, J= 7 Hz); 5.91 (1H, no d); 6.00 (1H, dt, J= 16 and 8 Hz); 6.35 (1H, d, J= 16 Hz); 6.51 (1H, no d); 6.80 (1H, d, J= 9 Hz); 6.87 (2H, d J = 9 Hz); 7.06 (2H, m); 7.26 (8H, complex).

LRMS (thermospray) m/z = 613 (MH+).

b) Solution of tert-butyl (3R,5E)-3-({[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]amino}carbonyl) -6-(3-methyl-4-phenoxyphenyl)hex-5-enoate (615 mg, 1.00 mmol), ammonium formate (310 mg, 4.9 mmol) and 10% palladium on carbon (65 mg) in methanol (8 ml) was stirred at 20 °C for 16 hours. The mixture was filtered through an Arbocel filter aid and concentrated under reduced pressure. The residue was dissolved in ethyl acetate and washed with saturated aqueous sodium chloride, dried (MgSO4) and concentrated under reduced pressure to give tert-butyl (3R)-3-({[(1S)-2,2-dimethyl- 1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]amino}carbonyl)-6-(3-methyl-4-phenoxyphenyl)hexanoate (550 mg, 90%) as a foam, which was transferred in the next step without purification.

Rf 0.5 (hexane:ethyl acetate = 3:1).

δH (300 MHz, CDCl 3 ) 1.03 (9H, s); 1.41 (9H, s); 1.33-1.60 (4H, m, shaded with other peaks); 1.50 (3H, d, J= 7 Hz); 2.16 (3H, s); 2.30 (1H, m); 2.48 (2H, m); 2.57 (2H, m); 4.26 (1H, d, J= 9 Hz); 5.10 (1H, pentet, J= 7 Hz); 6.22 (1H, no d); 6.44 (1H, no d); 6.77 (1H, d, J= 9 Hz); 6.86 (3H, m); 6.99 (2H, m); 7.26 (7H, complex).

LRMS (thermospray m/z = 615 (M+).

c) Trifluoroacetic acid (2.5 ml) was added dropwise over 5 minutes to a stirred solution of tert-butyl (3R)-3-({[(1S)-2,2-dimethyl-1-({[( 1R)-1-phenylethyl]amino}carbonyl)propyl]amino}carbonyl)-6-(3-methyl-4-phenoxyphenyl)hexanoate (550 mg, 0.9 mmol) in anhydrous dichloromethane (5 mL) under nitrogen at 20 °C. The solution was stirred for 1 hour and concentrated under reduced pressure. The residue was diluted with hydrochloric acid (2M) and extracted with three portions of ethyl acetate. The combined organic solutions were washed with saturated aqueous sodium chloride solution, dried (MgSO4) and concentrated under reduced pressure. Two doses of toluene were added and evaporated to give the title compound as a yellow resin (640 mg) stained with toluene and trifluoroacetic acid.

Rf 0.18 (dichloromethane:methanol = 99:1).

δH (300 MHz, CDCl 3 ) 0.99 (9H, s); 1.44 (3H, m); 1.50 (3H, d, J= 7 Hz); 1.57 (1H, m, ); 2.15 (3H, s); 2.43 (3H, m); 2.71 (2H, m); 4.43 (1H, d, J= 10 Hz); 5.06 (1H, pentet, J = 7 Hz); 6.77 (2H, m); 6.84 (2H, d, J = 8 Hz); 6.93 (1H, s); 7.00 (1H, t, J= 8 Hz); 7.10 (8H, complex); 7.46 (1H, d, J= 10 Hz); 7.74 (2H, no s).

LRMS (thermospray) m/z = 558 (M+).

Preparation 7

Methyl (2S,3R)-3-carboxy-2-ethoxy-hex-4-enoate

[image]

a) n-butyllithium (23.0 ml, 2.5 M in hexanes, 57.7 mmol) was added dropwise to a stirred solution of diisopropylamine (9.63 ml, 69.0 mmol) in anhydrous tetrahydrofuran (10 ml ) under nitrogen at -78 °C. The reaction vessel was then placed in an ice bath for 10 minutes, and cooled again to -78 °C. Anhydrous tetrahydrofuran (10 mL) was added, followed dropwise by a solution of (S)-diisopropyl malate (6.0 g, 27.0 mmol) in anhydrous tetrahydrofuran (20 mL). The mixture was allowed to warm to -20 °C and stirred for 8 hours. The mixture was then recooled to -78 °C and freshly distilled allyl iodide (2.75 mL, 30 mmol) was added dropwise. The mixture was allowed to warm to -40 °C and stirred for 36 hours. It was then poured into an ice-cold 5% aqueous solution of citric acid (150 ml) and extracted with ethyl acetate (3 x 150 ml). The combined organic solutions were washed with saturated aqueous sodium chloride solution, dried (MgSO4) and concentrated under reduced pressure. The residue was purified by flash chromatography (elution with hexane:ethyl acetate = 80:20) and a mixture of (2S,3R)- and (2S,3S)-isopropyl 2-hydroxy-3-(2-propyloxycarbonyl)-5-hexanoate was obtained. (5.42 g, 76 %) (14:1) as a pale yellow oil.

Rf 0.63 (hexane:ethyl acetate = 1:1).

δH (400 MHz, CDCl 3 ) (major isomer) 1.19 (6H, m); 1.26 (6H, m); 2.39 (1H, m); 2.58 (1H, m); 2.87 (1H, m); 3.14 (1H, d, J= 6.5 Hz, OH); 4.19 (1H, m); 4.98 (1H, m); 5.07 (2H, m); 5.13 (1H, d, J= 15 Hz); 5.79 (1H, m); (minor isomer peaks) 2.29 (1H, m); 2.50 (1H, m); 2.78 (1H, m); 3.06 (1H, d); 4.40 (1H, m).

LRMS (thermospray) m/z = 276 (MNH4+), 259 (MH+).

b) Ethyl trifluoromethanesulfonate (1.45 ml, 11.2 mmol) was added to a mixture of (2S,3R)- and (2S,3S)-isopropyl 2-hydroxy-3-(2-propyloxycarbonyl)-5-hexanoate (14: 1) (1.45 g, 5.6 mmol) and 2,6-di-tert-butyl-4-methylpyridine (2.88 g, 14.0 mmol) and the mixture was stirred under nitrogen at 45 °C 5, 5 hours, at 20 °C for 18 hours, and at 45 °C for 24 hours. The cooled mixture was placed in the upper part of a short column with silica gel and eluted with ethyl acetate. The filtrate was concentrated under reduced pressure and further purified by flash chromatography (hexane:ethyl acetate = 85:15), to give (2S,3R)-isopropyl 2-ethoxy-3-(2-propyloxycarbonyl)-5-hexenoate (627 mg , 39 %).

Rf 0.62 (hexane:ethyl acetate = 70:30).

δH (400 MHz, CDCl 3 ) 1.13 (3H, t, J = 7.5 Hz); 1.17 (6H, m); 1.24 (6H, m); 2.24 (1H, dt, J= 13 and 6.5 Hz); 2.36 (1H, dt, J= 13 and 6.5 Hz); 2.83 (1H, m); 3.38 (1H, dq, J= 9.5 and 7.5 Hz); 3.62 (1H, J= 9.5 and 7.5 Hz); 3.92 (1H, d, J= 8 Hz); 5.02 (4H, m); 5.70 (1H, m).

LRMS (thermospray) m/z 304 (MNH4+); 287 (MH+).

c) A mixture of (2S,3R)- and (2S,3S)- isopropyl 2-hydroxy-3-(2-propyloxycarbonyl)-5-hexanoate (500 mg, 1.75 mmol) and lithium hydroxide hydrate (154 mg, 3 .67 mmol) in methanol:water = 1:1 (6 ml) was stirred under nitrogen at room temperature for 22 hours and then concentrated under reduced pressure. NMR showed an incomplete reaction, so more lithium hydroxide hydrate (154 mg, 3.67 mmol) and methanol:water = 1:1 (6 mL) were added, and the mixture was heated at 50 °C for 20 h. The mixture was cooled, concentrated under reduced pressure, and toluene was added and evaporated several times. The residue was dried over grains of sodium hydroxide in a vacuum, and then it was used in the next step without purification.

δH (400 MHz, CD3OD) 1.15 (3H, t, J= 7.5 Hz); 2.26 (1H, m); 2.40 (1H, m); 2.46 (1H, m); 3.35 (1H, dq, J= 9.5 and 7.5 Hz); 3.45 (1H, dq, J= 9.5 and 7.5 Hz); 3.72 (1H, d, J= 7 Hz); 4.94 (2H, partially shaded with HOD peak); 5.84 (1H, m).

d) To a mixed suspension of dilithium (2S,3R)-2-ethoxy-3-(prop-3-enyl)butanedioate (393 mg, from c, above) in anhydrous tetrahydrofuran (2.6 ml) under nitrogen at 0 °C was added is trifluoroacetic anhydride (2.62 ml), drop by drop. The mixture was stirred for 4 hours and concentrated under reduced pressure. Toluene was added and evaporated several times. The resulting resin was dissolved in anhydrous methanol (2 ml) and allowed to warm to room temperature overnight. The solution was concentrated under reduced pressure, and the residue was partitioned into a saturated aqueous solution of sodium dihydrogen citrate and ethyl acetate. The aqueous layer was extracted with two portions of ethyl acetate. The combined organic solutions were washed with saturated aqueous sodium chloride, dried (Na 2 SO 4 ), and concentrated under reduced pressure to give the title compound (220 mg) as a pale yellow oil. One major impurity was identified via 1H NMR, which could be the (2S,3S) isomer.

Rf 0.3 (dichloromethane:methanol:acetic acid = 95:5:1).

δH (400 MHz, CD3OD) 1.16 (3H, t, J = 7.5 Hz); 2.32 (1H, m); 2.44 (1H, m); 2.95 (1H, m); 3.42 (1H, dq, J= 9.5 and 7.5 Hz); 3.68 (m); 3.74 (3H, s); 4.01 (1H, d, J= 6 Hz); 4.94 (2H, m); 5.74 (1H, m).

Example 36 / Preparation 8

(2S,3R)-3-([(1S)-2,2-dimethyl-1-([(1R)-1-phenylethyl]aminocarbonyl)propyl]aminocarbonyl)-2-ethoxy-6-(3-fluoro- 4-phenoxyphenyl)hexanoic acid

[image]

a) N-(dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (214 mg, 1.12 mmol) was added to a stirred mixture of methyl (2S,3R)-3-carboxy-2-ethoxy-hex-4-enoate (preparation 7) (210 mg, 0.97 mmol) and 1-hydroxy-7-azabenzotriazole (139 mg, 1.02 mmol) in anhydrous dimethylformamide (4 mL) under nitrogen at 0 °C. After 1 hour, (2S)-amino-3,3-dimethyl-N-[(1R)-1-phenylethyl]butanamide hydrochloride (preparation 1) (276 mg, 1.02 mmol) was added, followed by diisopropylethylamine (0 ,18 µl), 1.02 mmol). After another 30 minutes, the mixture was allowed to warm to room temperature. After 17 h at room temperature, the mixture was concentrated under reduced pressure and partitioned between ethyl acetate (50 ml) and water (50 ml). The aqueous layer was saturated with sodium bicarbonate and extracted with ethyl acetate (2 × 20 ml). The combined organic solutions were concentrated under reduced pressure, dissolved in ether (50 ml), washed with water (3 x 20 ml), dried (Na2SO4) and concentrated under reduced pressure. The residue was purified by flash chromatography (eluting with hexane:ethyl acetate = 60:40) and methyl(2S,3R)-3-({[(1S)-2,2-dimethyl-1-({[(1R)) was obtained -1-phenylethyl]amino}carbonyl)propyl]amino}carbonyl)-2-ethoxy-hex-5-enoate (181 mg, 43%) as a colorless solid (single diastereoisomer by 1H NMR).

T.t. 131-134 °C.

Rf 0.4 (hexane:ethyl acetate = 1:1).

δH (300 MHz, CDCl 3 ) 1.00 (9H, s); 1.22 (3H, t, J= 7 Hz); 1.49 (3H, d, J = 6.5 Hz); 2.31 (1H, dt, 14 and 8 Hz); 2.51 (1H, dt, 14 and 6.5 Hz); 2.68 (1H, m); 3.42 (1H, dq, J= 16 and 7 Hz); 3.66 (1H, dq, J= 16 and 7 Hz); 3.69 (3H, s); 4.05 (1H, d, J= 5 Hz); 4.13 (1H, d, J= 9 Hz); 5.04 (1H, d, J= 10 Hz); 5.07 (1H, d, J= 18 Hz); 5.14 (1H, pentet, J= 7 Hz); 5.70 (1H, dddd, J= 6, 7, 10 and 18 Hz); 6.42 (1H, no d); 7.06 (1H, no d); 7.29 (5H, complex).

LRMS (thermospray) m/z = 433 (MH+).

b) A mixture of palladium acetate (12 mg, 0.05 mmol) and tri-(2-methylphenyl)phosphine (30 mg, 0.10 mmol) in anhydrous acetonitrile (1 ml) was sonicated at room temperature for 1 min until no cream-colored suspension was formed. This suspension was added using a Pasteur pipette to a stirred solution of methyl (2S,3R)-3-({[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl) propyl]amino}carbonyl)-2-ethoxy-hex-5-enoate, (415 mg, 0.96 mmol), 3-fluoro-4-phenoxy-bromobenzene (preparation 10) (400 mg, 1.50 mmol) and of triethylamine (280 µl, 2.0 mmol) in anhydrous acetonitrile (1 ml) under nitrogen. The mixture was flushed with nitrogen and then heated at reflux for 16 hours. After cooling, the mixture was poured into ethyl acetate (70 ml) and washed with 5% aqueous citric acid (20 ml), saturated aqueous sodium chloride (50 ml), dried (MgSO4) and concentrated under reduced pressure. The residue was purified by flash chromatography (gradient elution with dichloromethane:ethyl acetate) and obtained mainly (2S,3R)-3-({[(1S)-2,2-dimethyl-1-({[(1R)-1- phenylethyl]amino}carbonyl)propyl]amino}carbonyl)-2-ethoxy-6-(3-fluoro-4-phenoxyphenyl)hex-5-enoate as a pale yellow foam (359 mg, 69 %). 1H NMR suggested that two alkene isomers, (5Z) and (4iE), were also present. The mixture of these two alkenes was carried to the next step (see c, below).

Rf 0.36 (dichloromethane:ethyl acetate = 10:1).

δH (400 MHz, CDCl3) (for (5E) isomer), 1.01 (9H, s); 1.16 (3H, t, J = 7 Hz); 1.49 (3H, d, J = 6.5 Hz); 2.48 (1H, m); 2.68 (1H, m); 2.80 (1H, m); 3.26 (1H, m); 3.69 (1H, m, and 3H, s, overlap); 4.06 (1H, d, J= 4 Hz); 4.15 (1H, d, J= 9.5 Hz); 5.10 (1H, pentet, J= 6.5 Hz); 6.03 (1H, dt, 16 and 8 Hz); 6.36 (1H, d, J= 16 Hz); 6.42 (1H, no d); 6.95 (3H, m); 7.01 (1H, d, J= 8 Hz); 7.09 (3H, m); 7.22 (1H, m); 7.28 (6H, complex).

LRMS (thermospray) m/z = 636 (MNH4+).

c) Solution of methyl (2S,3R,5E)-3-({[(1R)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]amino}carbonyl) -2-ethoxy-6-(3-fluoro-4-phenoxyphenyl)hex-5-enoate (350 mg, 0.566 mmol) in ethanol (15 ml) was hydrogenated over 10% palladium on carbon (60 mg) at 3 bar at 20 °C for 16 hours. The mixture was filtered through an Arbocel filter aid and concentrated under reduced pressure. The residue was dissolved in cyclohexane and evaporated (2×), to give methyl (2R,3S)-3-({[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl) amino}carbonyl)propyl]amino}carbonyl)-2-ethoxy-6-(3-fluoro-4-phenoxyphenyl)hexanoate (368 g, 105 %) as a colorless foam, which was carried to the next step without purification.

Rf 0.29 (hexane:ethyl acetate = 2:1).

δH (400 MHz, CDCl 3 ) 1.01 (9H, s); 1.19 (3H, t, J = 7 Hz); 1.49 (3H, d, J = 6.5 Hz); 1.61 (3H, m); 1.75 (1H, m); 2.58 (3H, m); 3.41 (1H, dq, J= 10 and 7 Hz); 3.61 (1H, dq, J= 10 and 7 Hz); 3.70 (3H, s); 3.99 (1H, d, J= 5 Hz); 4.15 (1H, d, J= 9 Hz); 5.12 (1H, pentet, J= 6.5 Hz); 6.38 (1H, no d); 6.83 (1H, d, J = 10 Hz); 6.87 (1H, d, J= 12 Hz); 6.93 (4H, m); 7.03 (1H, t, J = 9.5 Hz); 7.20 (1H, m); 7.29 (6H, complex).

LRMS (thermospray) m/z = 638 (MNH4+).

d) Lithium hydroxide hydrate (116 mg, 2.83 mmol) was added to a solution of methyl (2R,3S)-3-({[(1S)-2,2-dimethyl-1-({[(1R)-1- phenylethyl]amino}carbonyl)propyl]amino}carbonyl)-2-ethoxy-6-(3-fluoro-4-phenoxyphenyl)hexanoate (368 mg, 0.566 mmol) in methanol:water = 10:1 (5 ml), then the mixture was stirred at room temperature for 2 hours. The solution was neutralized with a 5% aqueous solution of citric acid and concentrated under reduced pressure. The residue was partitioned into ethyl acetate (20 mL), dried (MgSO 4 ) and concentrated under reduced pressure. Purification by flash chromatography (eluting with dichloromethane:methanol = 20:1 to 10:1) afforded the title compound (62 mg, 18%) as a colorless glass, and several mixed fractions. They were combined and purified again by flash chromatography (eluting with ethyl acetate:hexane:acetic acid = 33:66:1 to 50:50:1)), and the title compound (37 mg, 11 %) was obtained as a white solid matter.

T.t. 185-187 °C.

Rf 0.23 (ether:hexane:acetic acid = 70:30:1).

δH (400 MHz, CD3OD) (NB some chemical shifts depend on concentration) 1.02 (9H, s); 1.15 (3H, t, J= 7 Hz); 1.44 (3H, d, J = 7 Hz); 1.56 (3H, m); 1.69 (1H, m); 2.54 (2H, m); 2.68 (1H, m); 3.35 (1H, dq, J= 10 and 7 Hz); 3.66 (1H, dq, J= 10 and 7 Hz); 4.31 (1H, s); 5.02 (1H, q, J = 7 Hz); 6.87 (4H, m); 6.96 (1H, d, J= 12 Hz); 7.02 (1H, t, J= 7 Hz); 7.13 (1H, m); 7.19 (2H, t, J = 7 Hz); 7.28 (4H, m).

LRMS (thermospray) m/z = 607 (MH+).

FTIR νmax (KBr disc) 3330; 2980; 1643; 1593; 1508; 1493; 1217; 700 cm-1.

Preparation 9

(3-chloro-4-phenyl)phenyl trifluoromethanesulfonate

[image]

Trifluoromethanesulfonic anhydride (755 µl, 4.45 mmol) was added dropwise over 2 minutes to a stirred solution of (3-chloro-4-phenyl)phenol (759 mg, 3.71 mmol) (International Patent WO 97/20815) and anhydrous pyridine (758 µl, 9.65 mmol) in anhydrous dichloromethane under nitrogen at 0 °C. The mixture was stirred for 1.5 hours at 0 °C and then for 2.5 hours at 20 °C. The mixture was then poured into a mixture of ethane/hexane (200 ml, 50:50) and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by flash chromatography (eluting with hexane:ether = 20:1), and the title compound was obtained as a colorless oil (1062 mg, 85 %) which crystallized on standing at room temperature. couple of days.

T.t. 43-44 °C.

Rf 0.27 (hexane).

δH (300 MHz, CDCl3) 7.27 (1H, dd, J= 8 and 2 Hz); 7.29 (7H, no. s).

LRMS (EI) m/z = 336 (M+).

Preparation 10

3-fluoro-4-phenoxy-bromobenzene

[image]

Aqueous sodium hydroxide solution (5.5 mL, 1M, 5.5 mmol) was added to a mixture of 4-bromo-2-fluorophenol (0.96 g, 5.0 mmol) and diphenyliodonium bromide (1.99 g, 5.5 mmol) in water (20 ml), so the mixture was heated at reflux for 4 hours. After cooling, the reaction mixture was diluted with ether and filtered. The organic phase was washed with saturated aqueous sodium chloride solution, dried (MgSO4) and concentrated under reduced pressure. The remaining dark orange oil was purified by flash chromatography (eluting with hexane) to give the title compound as an orange oil (12 g, 90%).

Rf 0.36 (hexane).

δH (400 MHz, CDCl 3 ) 6.90 (3H, m); 7.06 (1H, m); 7.20 (1H, m); 7.30 (3H, m).

Preparation 11

1-iodo-3-methyl-4-phenoxybenzene

[image]

Aqueous sodium hydroxide solution (33 mL, 1M, 33 mmol) was added to a mixture of 4-iodo-2-methylphenol (7.0 g, 29.9 mmol) and diphenyliodonium bromide (11.89 g, 32.9 mmol) in water. (120 ml), and the mixture was heated under reflux for 4 hours. After cooling, the reaction mixture was extracted with ethyl acetate (4 x 100 ml), the combined extracts were washed with saturated aqueous sodium chloride solution, dried (MgSO4) and concentrated under reduced pressure. The remaining dark orange oil was purified by repeated flash chromatography (eluting with hexane in the first column and then pentane in the long column) to give the title compound as an orange oil (7.84 g, 84 %).

Rf 0.78 (hexane).

δH (400 MHz, CDCl 3 ) 2.23 (3H, s); 6.65 (1H, d, J= 9 Hz); 6.94 (2H, m); 7.10 (1H, m), 7.32 (2H, m), 7.47 (1H, d, J = 9 Hz), 7.59 (1H, s).

LRMS (thermospray) m/z = 310 (MH+).

Preparation 12

tert-butyl (2R,4R)-3-carboxy-2-propyl-hex-5-enoate

[image]

a) n-butyllithium (2.2 ml, 2.5 M in hexanes, 5.6 mmol) was added dropwise to a stirred solution of diisopropylamine (0.73 ml, 5.6 mmol) in anhydrous tetrahydrofuran (3 ml ) under nitrogen at -5 °C. After 1 hour, the solution was cooled to -70 °C and N,N'-dimethylpropyleneurea (DMPU) (3 ml) was added. A solution of (2R)-2-[2-(tert-butoxy)-2-oxoethyl]pent-4-enoic acid (535 mg, 2.5 mmol) in anhydrous tetrahydrofuran (6 ml) was then added dropwise. , for 15 minutes, maintaining the temperature at -70 °C. After stirring for 1 hour, 1-iodopropane (860 mg, 4.0 mmol) was added dropwise and the mixture was stirred at -70 °C for 1 hour, then at -40 °C for 2 hours, and at finally allowed to warm to 0 °C. Methanol (1 ml) was added, and the mixture was partitioned between 5% aqueous citric acid (150 ml) and ether (200 ml). The organic layer was washed with 5% aqueous citric acid solution (75 ml), water (3 x 200 ml), dried (MgSO4) and concentrated under reduced pressure to give a pale yellow oil. Purification by flash chromatography (eluting with hexane:ethyl acetate:acetic acid = 80:20:2) gave (2R,3R)- and (2R,3S)-3-[(2-methylprop-2-yl)oxocarbonyl]- 2-(3-propen-1-yl)hexanoic acid (472 mg, 74%), in the form of a colorless oil with a diastereometric ratio of 97:3.

Rf 0.31 (hexane:ethyl acetate:acetic acid = 80:20:2).

δH (400 MHz, CDCl3) (for 2R,3R isomer) 0.92 (3H, t, J= 7.5 Hz); 1.34 (2H, m); 1.45 (9H, s); 1.58 (2H, m); 2.39 (2H, t, J = 7 Hz); 2.60 (1H, dt, J= 6 and 8 Hz); 2.74 (1H, dt, 6 and 7.5 Hz); 5.06 (1H, d, J=10 Hz); 5.10 (1H, d, J=17 Hz); 5.74 (1H, ddt, J=17, 10 and 7.5 Hz).

LRMS (thermospray) m/z = 256 (weak, M+); 200 (base peak, MH+ - t-Bu).

FTIR νmax (film) 2970; 1730; 1370; 1250; 1160 cm-1.

b) Mixture of (2R,3R)- and (2R,3S)-3-[(2-methylprop-2-yl)oxocarbonyl]-2-(3-propen-1-yl)hexanoic acid (97:3 from a , above) (3.05 g, 11.9 mmol) was dissolved in anhydrous tetrahydrofuran (35 mL) and added dropwise over 30 min to a stirred solution of lithium diisopropylamide (from n-butyllithium) (19.2 mL, 2 .5 M in hexanes, 48 mmol) and diisopropylamine (7.0 ml, 50 mmol) in anhydrous tetrahydrofuran (25 ml) at -70 °C. The mixture was allowed to warm to -10 °C, stirred for 30 minutes, and then allowed to warm to 0 °C for 2 hours. The solution was then cooled to -70 °C and methanol (15 ml) was quickly added using a syringe. The mixture was allowed to warm to room temperature and then partitioned into a 5% aqueous solution of citric acid (150 ml) and ether (200 ml). The organic layer was washed with 5% aqueous citric acid solution (50 ml), water (3 x 200 ml) and concentrated under reduced pressure to give a pale yellow oil. Purification by flash chromatography (gradient elution with hexane:ethyl acetate:acetic acid) gave (2R,3R)- and (2R,3S)- 3-[(2-methylprop-2-yl)oxocarbonyl]-2-(3- propen-1-yl)hexanoic acid in the form of a colorless oil with a diastereometric ratio of 58:42.

Rf 0.31 (hexane:ethyl acetate:acetic acid) = 80:20:2).

The stereochemistry of the two isomers was determined as follows. A mixture of (2R,3R)- and (2R,3S)-3-[(2-methylprop-2-yl)oxocarbonyl]-2-(3-propen-1-yl)hexanoic acid (ratio 1:3) in ethanol was hydrogenated over 10% palladium on carbon (3 bar), then treated with hydrogen chloride in dry dichloromethane/dioxane (4 °C, 4 hours) and a mixture of (2R,3S)- and (2R,3R) was obtained - 2,3-diprop-1-yl butanedioic acids (ratio 1:3), which are known from the literature ("Bull. Chem.", France, 2189, (1975)). Examination of the proton NMR spectrum (acetone-d6) gave δH 2.70 (2H, m, CHCO2H, major isomer) and δH 2.55 (2H, m, CHCO2H, minor isomer).

c) A solution of (2R,3R)- and (2R,3S)-3-[(2-methylprop-2-yl)oxocarbonyl]-2-(3-propen-1-yl)hexanoic acid (3.04 g, 11.9 mmol) (diastereometric ratio 58:42) and cesium bicarbonate (2.30 g, 11.9 mmol) in water (35 ml) and acetonitrile (90 ml) was stirred at 20 °C for 5 min. The solution was then evaporated under reduced pressure and the residue was dissolved and evaporated from toluene 4 times. The resulting resin was dried under high vacuum for 30 minutes and then dissolved in anhydrous N,N'-dimethylacetamide (50 ml). Iodomethane (3.41 g, 24 mmol) was added and the solution was stirred at 20 °C for 17 h. The solution was concentrated under reduced pressure and partitioned between ether (250 ml) and water (250 ml). The organic layer was washed with water (3 × 200 ml), dried (MgSO4) and concentrated under reduced pressure, after which a mixture of methyl (2R,3S)- and (2R,3R)-3-[(2-methylprop-2) was obtained -yl)oxycarbonyl]-2-(3-propen-1-yl)hexanoate as a yellow oil (3.0 g, 94 %). TLC (pentane:ether = 10:1) showed two product spots with Rf 0.41 and 0.30.

d) A mixture of (2R,3S)- and (2R,3R)-3-[(2-methylprop-2-yl)oxycarbonyl]-2-(3-propen-1-yl)hexanoate (from c, above), it was separated by flash chromatography (pentane:ether = 20:1), after which two fractions were obtained. The first eluted product was identified as (2R,3S)-3-[(2-methylprop-2-yl)oxycarbonyl]-2-(3-propen-1-yl)hexanoate (1.78 g, 56 %, Rf 0 ,41 (pentane:ether = 10:1) in the form of a colorless oil.

δH (400 MHz, CDCl 3 ) 0.87 (3H, t, J= 7 Hz); 1.30 (3H, m); 1.46 (9H, s); 1.56 (1H, m); 2.23 (1H, m); 2.34 (1H, m); 2.54 (1H, dt, 3 and 10 Hz); 2.69 (1H, 4.5 and 10 Hz); 3.66 (3H, s); 5.00 (1H, d, J=10 Hz); 5.04 (1H, d, J=17 Hz); 5.72 (1H, ddt, J=17, 10 and 7 Hz).

Found: C, 66.64; H, 9.69.

C15H28O4 required: C, 66.61; H, 9.77%.

FTIR νmax (KBr disk) 2970; 1730; 1370; 1150 cm-1.

The second eluted product was identified as (2R,3R)-3-[(2-methylprop-2-yl)oxycarbonyl]-2-(3-propen-1-yl)hexanoate (760 mg, 24 %, Rf 0.30 (pentane:ether = 10:1) in the form of a colorless oil.

δH (400 MHz, CDCl 3 ) 0.90 (3H, t, J= 7 Hz); 1.27 (1H, m); 1.35 (1H, m); 1.45 (9H, s); 1.55 (2H, m); 2.36 (2H, m); 2.58 (1H, dt, J= 5 and 9 Hz); 2.71 (1H, dt, 5 and 9 Hz); 3.66 (3H, s); 5.02 (1H, d, J=10 Hz); 5.08 (1H, d, J=16 Hz); 5.76 (1H, m).

Found: C, 66.64; H, 9.69.

C15H26O4 required: C, 66.70; H, 9.81%.

FTIR νmax (KBr disk) 2970, 1730, 1370, 1160 cm-1.

e) A mixture of (2R,3S)-3-[(2-methylprop-2-yl)oxycarbonyl]-2-(3-propen-1-yl)hexanoate (1.49 g, 5.51 mmol) and anhydrous lithium of iodide (11 g, 82 mmol) in anhydrous pyridine (50 mL) was heated at reflux under nitrogen for 17 h. After cooling, the mixture was poured into a 20% aqueous solution of citric acid (500 ml) and extracted with ethyl acetate (250 ml). The organic layer was washed with 5% aqueous citric acid solution (2 x 150 ml), water (3 x 250 ml), dried (MgSO4) and concentrated under reduced pressure. The residue was triturated with hexane and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by repeated flash chromatography (the first column was eluted with dichloromethane:ethyl acetate:acetic acid = 80:20:2, and the second with hexane:ethyl acetate:acetic acid, gradient) and (2R,3S)-3- [(2-Methylprop-2-yl)oxycarbonyl]-2-(3-propen-1-yl)hexanoic acid (788 mg, 56%) as a colorless oil.

Rf 0.3 (hexane:ethyl acetate:acetic acid = 80:20:2).

δH (400 MHz, CDCl 3 ) 0.91 (3H, t, J= 7 Hz); 1.35 (3H, m); 1.47 (9H, s); 1.62 (1H, m); 2.26 (1H, dt, J=10 and 7.5 Hz); 2.38 (1H, dt, J= 7.5 and 10 Hz); 2.58 (1H, dt, J= 3 and 10 Hz); 2.70 (1H, dt, 5 and 9 Hz); 5.04 (1H, d, J=10 Hz); 5.08 (1H, d, J=18 Hz); 5.76 (1H, ddt, J= 18, 10 and 7.5 Hz).

LRMS (thermospray) m/z = 257 (weak, MH+).

Found: C, 65.30; H, 9.44.

C14H24O4 required: C, 65.60; H, 9.44%.

Preparation 13

tert-butyl (3R)-3-({[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]amino}carbonyl)-6-[3 -methyl-(4-phenyl)phenyl]-(2S)-propylhexanoate

[image]

a) According to the procedure for preparation 2, (2R,3S)-3-[(2-methylprop-2-yl)oxycarbonyl]-2-(3-propen-1-yl)hexanoic acid (380 mg, 1.48 mmol) was reacted with (2S)-amino-3,3-dimethyl-N-[(1R)-1-phenylethyl]butanamide hydrochloride (preparation 1) (441 mg, 1.62 mmol) for 1 hour at 4 °C, and then for 72 hours at 20 °C. The mixture was concentrated under reduced pressure and partitioned between ethyl acetate (100 ml) and water (100 ml). The aqueous layer was saturated with sodium bicarbonate and extracted with ethyl acetate (2 x 100 ml). The combined organic solutions were concentrated under reduced pressure and the residue was dissolved in ether (500 ml), washed with water (3 x 200 ml), dried (MgSO4) and concentrated under reduced pressure. The residue was triturated with pentane and dried, after which tert-butyl (2S,3R)-3-({[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl)]amino was obtained }carbonyl)propyl]amino}carbonyl)-2-(1-propyl)hex-5-enoate (579 mg, 80%), as a colorless solid.

T.t. 180-183 °C.

Rf 0.51 (hexane:isopropanol = 90:10).

δH (400 MHz, CDCl 3 ) 0.84 (3H, t, J = 7.5 Hz); 1.02 (9H, s); 1.22-1.50 (4H, m); 1.45 (9H, s); 1.50 (3H, d, J= 7 Hz); 2.08 (1H, m); 2.22 (1H, m); 2.34 (1H, dt, J= 4 and 10 Hz); 2.52 (1H, dt, J= 3 and 10 Hz); 4.18 (1H, d, J= 9 Hz); 4.72 (1H, d, J = 9.5 Hz); 4.85 (1H, d, J=16.5 Hz); 5.09 (1H, pentet, J = 7 Hz); 5.53 (1H, ddt, J= 9.5, 16.5 and 7 Hz); 6.03 (1H, no d); 6.24 (1H, no d); 7.26 (5H, complex).

LRMS (thermospray) m/z = 473 (MH+).

FTIR νmax (KBr disc) 3320; 2970; 2930; 1725; 1640; 1540; 1370; 1150; 700 cm-1.

Found: C, 71.09; H, 9.46; N, 6,10.

C28H44N2O4 required: C, 71.15; H, 9.38; N, 5.93%.

b) According to the preparation procedure 3, tert-butyl (2S,3R)-3-({[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl ]amino}carbonyl)-2-(1-propyl)hex-5-enoate (543 mg, 1.15 mmol) was reacted with 3-methyl-4-phenylbromobenzene (356 mg, 1.44 mmol) under catalysis palladium in a mixture of anhydrous acetonitrile and dimethylformamide (2:5, 7 ml) at 90 °C for 17 hours. The mixture was then concentrated under reduced pressure, poured into ethyl acetate (100 ml), washed with water (2 x 100 ml), dried (MgSO 4 ) and concentrated under reduced pressure. The residue was purified by flash chromatography (gradient elution with hexane:ethyl acetate) and mainly tert-butyl (2S,3R,5E)-3-({[(1S)-2,2-dimethyl-1-({{( 1R)-1-phenylethyl]amino}carbonyl)propyl]amino}carbonyl)-6-[3-methyl-(4-phenyl)phenyl]-2-(1-propyl) hex-5-enoate as a colorless resin (460 mg, 63 %). 1H NMR suggested that two alkene isomers, (5Z) and (4iE), were also present. The alkene mixture was carried to the next step (see c, below).

Rf 0.46 (hexane:ethyl acetate = 3:1).

δH (400 MHz, CDCl3) (for (5E) isomer) 0.87 (3H, t, J= 7 Hz); 1.03 (9H, s); 1.32 (2H, m); 1.46 (9H, s, 3H, d, J= 7 Hz, and 2H, m, overlap); 2.22 (3H, s); 2.34 (1H, m); 2.48 (2H, m); 2.60 (1H, m); 4.19 (1H, d, J= 9 Hz); 4.93 (1H, pentet, J= 7 Hz); 5.85 (1H, no d); 6.05 (1H, dt, J=14 and 7 Hz); 6.35 (1H, d, J=14 Hz); 6.27 (1H, no d); 7.20 (11H, complex); 7.37 (2H, m).

LRMS (thermospray) m/z = 639 (MH+).

c) tert-butyl (2S,3R,5E)-3-({[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]amino}carbonyl )-6-[3-methyl-(4-phenyl)phenyl]-2-(1-propyl)hex-5-enoate (437 mg, 0.68 mmol) in ethanol (50 ml) was hydrogenated over 10%- of palladium on carbon (50 mg) at 3 bar at 20 °C for 17 hours. The mixture was filtered through an Arbocel filter aid and concentrated under reduced pressure. Flash chromatography (gradient elution with hexane:ethyl acetate) gave (2S,3R)-3-({[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl) )propyl]amino}carbonyl)-6-[3-methyl-(4-phenyl)phenyl]-2-(1-propyl)hexanoate (395 g, 62%), as a colorless foam.

Rf 0.38 (hexane:ethyl acetate = 3:1).

δH (400 MHz, CDCl 3 ) 0.85 (3H, t, J= 7 Hz); 1.03 (9H, s); 1.26 (4H, m); 1.37 (9H, s); 1.48 (4H, m, and 3H, d, J= 7 Hz, overlap); 2.20 (3H, s); 2.29 (1H, m); 2.48 (4H, m); 4.24 (1H, d, J= 9 Hz); 5.10 (1H, pentet, J= 7 Hz); 6.02 (1H, no d); 6.29 (1H, no d); 6.94 (1H, d, J= 8 Hz); 6.98 (1H, s); 7.09 (1H, d, J= 8 Hz); 7.24 (7H, complex); 7.32 (1H, d, J= 7 Hz); 7.39 (2H, t, J = 7 Hz).

LRMS (thermospray) m/z = 641 (MH+).

Found: C, 76.71; H, 8.97; N, 4.35.

C41H56N2O4 required: C, 76.84; H, 8.81; N, 4.37%.

Preparation 14

(2R)-2-[(4S)-2,2-dimethyl-5-oxo-1,3-dioxolan-4-yl]-N-[(1S)-2,2-dimethyl-1-({[ (1R)-1-Phenylethyl]amino}carbonyl)propyl]pent-4-enamide

[image]

a) A mixture of isopropyl (2S,3R)- and (2S,3S)-2-hydroxy-3-(2-propyloxycarbonyl)-5-hexenoate (6.28 g, 24.3 mmol) (isomeric ratio = 14:1 ) (preparation 7, step a) and potassium hydroxide (4.09 g, 72.9 mmol) in dioxane:water (50 ml, 5:2) was heated under nitrogen at 90 °C for 18 h. The mixture was cooled, diluted with water (200 ml) and passed through an ion-exchange column (Dowex 50X8, 300 g) eluting with water until diacid elution stopped. The eluant was concentrated under reduced pressure, dissolved and evaporated with ethanol 2 times, from ether 2 times, and dried in vacuum, after which (2S,3R)-2-hydroxy-3-(prop-3-en-1- il)butanedioic acid (waxy yellow solid, 4.85 g, >100%, with ethanol and ether content), as the main component of the 8:1 mixture of isomers, which was used directly in the next step without further purification.

δH (300 MHz, CDCl 3 ) 2.47 (1H, dt, J= 7.5 and 15.5 Hz); 2.60 (1H, m); 3.00 (1H, dt, J= 4 and 6 Hz); 4.26 (1H, d, J= 4 Hz); 5.09 (1H, d, J=10 Hz); 5.15 (1H, d, J=18 Hz); 5.83 (1H, m).

b) The crude diacid from a, above, was dissolved in a mixture of dimethylformamide (35 ml) and 2,2-dimethoxypropane (133 ml) at room temperature, and p-toluenesulfonic acid hydrate (231 mg) was added. The mixture was heated at 30 °C for 2.5 days. The solvents were removed under reduced pressure to give 7.5 g of an oil containing (4S)-4-[(1R)-1-carboxy-but-3-enyl]-2,2-dimethyl-1,3- dioxolan-5-one, as the main component, together with dimethylformamide and p-toluenesulfonic acid.

δH (400 MHz, CDCl 3 ) 1.53 (3H, s); 1.60 (3H, s); 2.47 (1H, dt, J= 7.5 and 15.5 Hz); 2.71 (1H, dt, J= 7.5 and 15.5 Hz); 3.00 (1H, m); 4.30 (1H, d, J= 4 Hz); 5.12 (1H, d, J=10 Hz); 5.16 (1H, d, J=16 Hz); 5.80 (1H, m).

c) Part of the crude (4S)-4-[(1R)-1-carboxy-but-3-enyl]-2,2-dimethyl-1,3-dioxolan-5-one from b, above, (calculated from NMR so that it contains 1.54 g, 7.19 mmol), and 1-hydroxy-7-aza-1H-1,2,3-benzotriazole (1.09 g, 7.55 mmol) in anhydrous dichloromethane (30 ml) were mixed and cooled to 0 °C under nitrogen. N-(dimethylpropyl)-N'-ethylcarbodiimide hydrochloride (1.60 g, 8.27 mmol) was added and the mixture was stirred for 1.5 h at 0 °C. (2S)-amino-3,3-dimethyl-N-[(1R)-1-phenylethyl]butanamide hydrochloride (preparation 1) (2.15 g, 7.55 mmol) was then added, followed by diisopropylethylamine (1, 37 ml, 7.55 mmol). After 45 minutes, the mixture was allowed to warm to room temperature. After 3 hours at this temperature, the mixture was partitioned between ethyl acetate (200 ml) and pH 7 aqueous phosphate buffer (100 ml). The organic layer was washed with sodium bicarbonate, with salt, dried (Na2SO4), and concentrated under reduced pressure. The resulting foam crystallized upon addition of ether, and a white solid (1.89 g) was filtered off. Recrystallization from ethyl acetate:hexane gave (2R)-2-[(4S)-2,2-dimethyl-5-oxo-1,3-dioxolan-4-yl]-N-[(1S)-2,2 -dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]pent-4-enamide, as a white solid (1.30 g, 42%, as the only diastereomer).

T.t. 178-180 °C.

Rf 0.25 (dichloromethane: ether = 90:10).

δH (300 MHz, CDCl 3 ) 1.02 (9H, s); 1.48 (3H, d, J = 7 Hz); 1.51 (3H, s); 1.56 (3H, s); 2.47 (1H, dt, 15 and 7.5 Hz); 2.58 (1H, dt, 15 and 7.5 Hz); 2.70 (1H, dt, J= 11 and 5 Hz); 4.15 (1H, d, J= 9 Hz); 4.50 (1H, d, J= 5 Hz); 4.99 (1H, d, J=10 Hz); 5.08 (2H, m); 5.67 (1H, m, ); 5.67 (1H, no d); 6.45 (1H, no d); 7.27 (5H, complex).

LRMS (thermospray) m/z = 431 (MH+).

Preparation 15

(2R)-2-[(4S)-2,2-dimethyl-5-oxo-1,3-dioxolan-4-yl]-N-[(1S)-2,2-dimethyl-1-({[ (1R)-1-phenylethyl]amino}carbonyl)propyl]-5-[(3-fluoro-4-phenoxy)phenyl]pentanamide

[image]

a) A mixture of palladium acetate (17 mg, 0.075 mmol) and tri-(2-methylphenyl)phosphine (46 mg, 0.15 mmol) in anhydrous acetonitrile (2 ml) was sonicated at room temperature for 1 minute until formation cream-colored suspension. This suspension was added with a Pasteur pipette to a stirred solution of (2R)-2-[(4S)-2,2-dimethyl-5-oxo-1,3-dioxolan-4-yl]-N-[(1S)-2 ,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]pent-4-enamide (640 mg, 1.5 mmol), 1-bromo-3-fluoro-4-phenoxybenzene (preparation 10) (587 mg, 2.26 mmol) and N-ethylmorpholine (348 µl, 3.0 mmol) in anhydrous acetonitrile (2.5 ml) under nitrogen. The mixture was flushed with nitrogen and then heated at reflux for 16 hours. After cooling, the mixture was poured into ethyl acetate (100 ml) and washed with 5% aqueous citric acid (30 ml), saturated aqueous sodium chloride (50 ml), dried (Na 2 SO 4 ), and concentrated under reduced pressure. The residue was purified by flash chromatography (gradient elution with dichloromethane:ether) to give mainly (2R,5E)-2-[(4S)-2,2-dimethyl-5-oxo-1,3-dioxolan-4-yl] -N-[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]-5-(3-fluoro-4-phenoxyphenyl)pent-4-enamide , in the form of a pale yellow foam (210 mg, 22 %). 1H suggests that alkene isomers were also present. The alkene mixture is transferred to the next step (see b, below).

Rf 0.36 (dichloromethane.ether = 10:1).

δH (400 MHz, CDCl 3 ) (for (5E) isomer) 1.00 (9H, s); 1.50 (3H, d, J= 7 Hz); 1.53 (3H, s); 1.58 (3H, s); 2.65 (1H, m); 2.77 (2H, m); 4.18 (1H, d, J= 9 Hz); 4.56 (1H, d, J= 5 Hz); 5.03 (1H, pentet, J= 7 Hz); 5.88 (1H, no d); 6.01 (1H, dt, 16 and 7 Hz); 6.43 (1H, d, J=16 Hz); 6.58 (1H, no d); 6.94 (4H, m); 7.08 (2H, m); 7.20 (2H, m); 7.24 (5H, complex).

LRMS (thermospray) m/z = 618 (MH+); 542 (M+ - (CH3)2CO2).

b) Solution of (2R,5E)-2-[(4S)-2,2-dimethyl-5-oxo-1,3-dioxolan-4-yl]-N-[(1S)-2,2-dimethyl- 1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]-5-(3-fluoro-4-phenoxyphenyl)pent-4-enamide (210 mg, 0.34 mmol) in ethanol:ethyl acetate = 5:1 (10 ml) was hydrogenated over 10% palladium on carbon (20 mg) at 3 bar at 20 °C for 3 hours. The mixture was filtered through Arbocel filter aid and concentrated under reduced pressure to give (2R)-2-[(4S)-2,2-dimethyl-5-oxo-1,3-dioxolan-4-yl]-N- [(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]-5-(3-fluoro-4-phenoxyphenyl)pentanamide (103 mg, 49 %) in the form of a colorless solid, which was used in Example 9 without purification.

Rf 0.71 (dichloromethane: methanol = 90:10).

δH (400 MHz, CDCl 3 ) 1.03 (9H, s); 1.50 (3H, d, J= 7 Hz); 1.52 (3H, s); 1.57 (3H, s, and 2H, m, overlap); 1.85 (2H, m); 2.58 (3H, m); 4.16 (1H, d, J=10 Hz); 4.45 (1H, d, J= 8 Hz); 5.06 (1H, pentet, J = 7 Hz); 5.92 (1H, no d); 6.55 (1H, no d,); 6.84 (1H, d, J= 8 Hz); 6.94 (3H, m); 7.06 (1H, t, J = 7 Hz); 7.26 (8H, complex).

LRMS (thermospray) m/z = 619 (MH+).

Preparation 16

(2R)-2-[(4S)-2,2-dimethyl-5-oxo-1,3-dioxolan-4-yl]-N-[(1S)-2,2-dimethyl-1-({[ (1R)-1-phenylethyl]amino}carbonyl)propyl]-5-[(3-methyl-4-phenyl)phenyl]pentanamide

[image]

a) According to the procedure for making preparation 15a, (2R)-2-[(4S)-2,2-dimethyl-5-oxo-1,3-dioxolan-4-yl]-N-[(1S)-2, 2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]pent-4-enamide (preparation 14) (860 mg, 2.0 mmol) was reacted with 1-bromo- with 3-methyl-4-phenylbenzene (740 mg, 3.0 mmol) under palladium catalysis at reflux in acetonitrile for 16 h. After cooling, the mixture was partitioned between ethyl acetate (3 x 50 ml) and saturated aqueous sodium bicarbonate (50 ml). The combined organic solutions were dried (Na2SO4) and concentrated under reduced pressure. The residue was purified by flash chromatography (gradient elution with dichloromethane:methanol = 97.5:2.5) and obtained mainly (2R,5E)-2-[(4S)-2,2-dimethyl-5-oxo-1, 3-dioxolan-4-yl]-N-[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]-5-[(3-methyl- 4-phenyl)phenyl]pent-4-enamide, in the form of a brown foam (480 mg, 40 %). 1H NMR suggested that alkene isomers were also present. The alkene mixture was carried to the next step (see b, below).

Rf 0.59 (dichloromethane:methanol = 95:5).

δH (400 MHz, CDCl3) (for (5E) isomer) 1.03 (9H, s); 1.48 (3H, d, J = 7 Hz); 1.54 (3H, s); 1.59 (3H, s); 2.25 (3H, s); 2.71 (1H, dt, J= 15 and 7.5 Hz); 2.77 (1H, q, J= 6 Hz); 2.85 (1H, dt, J=15 and 7.5 Hz); 4.18 (1H, d, J=9Hz); 4.58 (1H,d, J=5Hz); 5.03 (1H, pentet, J=7 Hz); 5.87 (1H, no d); 6.13 (1H, dt, 16 and 7 Hz); 6.52 (1H, d, J=16 Hz); 6.58 (1H, no d); 7.23 (10H, complex); 7.38 (3H, m).

LRMS (thermospray) m/z = 521 (M+ - (CH3)2CO2).

b) Solution of (2R,5E)-2-[(4S)-2,2-dimethyl-5-oxo-1,3-dioxolan-4-yl]-N-[(1S)-2,2-dimethyl- 1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]-5-[(3-methyl-4-phenyl)phenyl]pent-4-enamide (480 mg, 0.80 mmol) in ethanol (20 ml) was hydrogenated over 10% palladium on carbon (48 mg) at 3 bar at 20 °C for 4 h. The mixture was filtered through Arbocel filter aid and concentrated under reduced pressure to give (2R)-2-[(4'S)-2,2-dimethyl-oxo-1,3-dioxolan-4-yl]-N-[( 1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]-5-[(3-methyl-4-phenyl)phenyl]pentathanamide (440 mg, 92 % ) in the form of a cream-colored solid, which was used in example 10 without purification.

Rf 0.53 (dichloromethane:methanol = 95:5).

δH (400 MHz, CDCl 3 ) 1.03 (9H, s); 1.50 (3H, d, J= 7 Hz); 1.53 (3H, s); 1.58 (3H, s); 1.64 (2H m); 1.87 (2H, m); 2.46 (3H, m); 4.20 (1H, d, J=10 Hz); 4.47 (1H, d, J= 5 Hz); 5.07 (1H, pentet, J= 7 Hz); 5.99 (1H, no d); 6.55 (1H, no d,); 7.00 (1H, d, J= 8 Hz); 7.04 (1H, s); 7.13 (1H, d, J= 8 Hz); 7.29 (5H, complex); 7.40 (2H, m).

LRMS (thermospray) m/z = 600 (MH+); 523 (base peak, M+ - (CH3)2CO2).

Example 37 / Preparation 17

(3R)-3-({[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]amino}carbonyl)-6-[(3-fluoro -4-phenyl)phenyl]hexanoic acid

[image]

Preparation 3 procedures were used for step a (using 1-bromo-3-fluoro-4-phenylbenzene instead of 1-bromo-methyl-4-phenylbenzene) and b to obtain:

tert-butyl (3R,5E)-3-({[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]amino}carbonyl)-6- [3-Fluoro-(4-phenyl)phenyl]hex-5-enoate as a colorless resin (997 mg, 83 %). 1H NMR suggested that two alkene isomers, (5Z) and (4E), were also present. The alkene mixture was transferred to the next step.

Rf 0.23 (hexane:ethyl acetate = 3:1).

δH (400 MHz, CDCl3) (for (5E) isomer) 0.96 (9H, s); 1.39 (9H, s); 1.47 (3H, d, J = 6.5 Hz); 2.28 (1H, dt, J= 14 and 7 Hz); 2.37 (1H, dd, J= 4 and 15 Hz); 2.44 (1H, m); 2.62 (1H, dd, J= 8 and 15 Hz); 2.73 (1H, m); 4.16 (1H, d, J= 9 Hz); 5.03 (1H, pentet, J= 6.5 Hz); 5.88 (1H, no d); 6.06 (1H, dt, J= 7.5 and 15 Hz); 6.33 (1H, d, J=15 Hz); 6.48 (1H, no d); 7.06 (2H, m); 7.19 (5H, m); 7.30 (2H, m); 7.39 (2H, t, J = 8 Hz); 7.48 (2H, d, J= 8 Hz).

LRMS (thermospray) m/z = 602 (MH+);

and

tert-butyl (3R)-3-({[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]amino}carbonyl)-6-[3 -fluoro-(4-phenyl)phenyl]hexanoate (702 mg, 72%), in the form of a colorless foam.

Rf 0.28 (hexane:ethyl acetate = 3:1).

δH (400 MHz, CDCl 3 ) 1.00 (9H, s); 1.37 (9H, s); 1.46 (3H, d, J = 7 Hz); 1.50 (3H, m); 1.61 (1H, m); 2.28 (1H, m); 2.55 (4H, m); 4.16 (1H, d, J= 9 Hz); 5.06 (1H, pentet, J = 7 Hz); 5.90 (1H, no d); 6.42 (1H, no d); 6.89 (2H, m); 7.22 (7H, complex); 7.40 (2H, t, J = 7 Hz); 7.48 (2H, d, J = 7 Hz).

LRMS (thermospray) m/z = 603 (MH+).

Found: C, 73.73; H, 7.86; N, 4.65.

C37H47FN2O4 required: C, 73.66; H, 7.88; N, 4.64%.

c) According to the procedures for making preparation 1, tert-butyl (3R)-3-({[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl] amino}carbonyl)-6-[3-fluoro-(4-phenyl)phenyl]hexanoate (650 mg, 1.08 mmol) was treated with trifluoroacetic acid at 20 °C for 4 hours to give the title compound (470 mg, 80 %).

T.t. 165-169 °C (after trituration with diisopropyl ether).

Rf 0.38 (ether:hexane:acetic acid = 70:30:1).

δH (400 MHz, DMSO-d6) 0.87 (9H, s); 1.43 (3H, d, J = 7 Hz); 1.42 (4H, m); 2.20 (1H, dd, J= 3 and 15 Hz); 2.44 (3H, m); 2.79 (1H, m); 4.27 (1H, d, J=10 Hz); 4.89 (1H, pentet, J = 7 Hz); 6.97 (2H, m); 7.16 (5H, complex); 7.32 (2H, m); 7.45 (4H, m); 7.65 (1H, no d); 8.27 (1H, no d); 11.97 (1H, no s).

Found: C, 72.50; H, 7.19; N, 5,12.

C33H39FN2O4 required: C, 72.52; H, 7.24; N, 5.09%.

FTIR νmax (KBr disc) 3300; 3070; 3030; 2965; 1710; 1633; 1543; 764; 700 cm-1.

Preparation 18

methyl (3R)-3-({[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]amino}carbonyl)-(2S)-ethoxy- 6-[(3-methyl-4-phenyl)phenyl]hexanoate

[image]

a) According to the procedure for making preparation 15a, methyl (2S,3R)-3-({[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl] amino}carbonyl)-2-ethoxy-hex-5-enoate (415 mg, 0.96 mmol) (preparation 8a) (396 mg, 0.92 mmol), was reacted with 1-bromo-3-methyl- with 4-phenylbenzene (340 mg, 1.37 mmol) under palladium catalysis at reflux in acetonitrile for 16 hours. After cooling, the mixture was poured into ethyl acetate (70 ml) and washed with 5% aqueous citric acid (20 ml), saturated aqueous sodium chloride (50 ml), dried (MgSO4) and concentrated under reduced pressure. The residue was purified by flash chromatography (gradient elution with hexane:ethyl acetate) and obtained mainly methyl (2S,3R,5E)-3-({[(1S)-3-({[(1S)-2,2- dimethyl-1-((1R)-1-phenylethyl]amino}carbonyl)propyl]amino}carbonyl)-2-ethoxy-6-[(3-methyl-4-phenyl)phenyl]hex-5-enoate as pale yellow foam (415 mg, 75%). 1H NMR suggested that two alkene isomers, (5Z) and (4E), were also present. The alkene mixture was carried to the next step (see b, below).

Rf 0.51 (hexane:ethyl acetate = 1:1).

δH (400 MHz, CDCl3) (for (5E) isomer) 1.02 (9H, s); 1.26 (3H, t, J = 7 Hz); 1.50 (3H, d, J= 7 Hz); 2.24 (3H, s); 2.55 (1H, m); 2.73 (1H, m); 2.82 (1H, m); 3.45 (1H, pentet, J= 7 Hz); 3.69 (1H, m, and 3H, s, overlap); 4.11 (1H, d, J= 4 Hz); 4.14 (1H, d, J=10 Hz); 5.12 (1H, pentet, J = 7 Hz); 6.14 (1H, dt, 16 and 8 Hz); 6.46 (1H, d, J=16 Hz); 6.50 (1H, no d); 7.24 (11H, complex); 7.40 (3H, m).

b) Solution of methyl(2S,3R,5E)-3-({[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]amino}carbonyl) -2-ethoxy-6-[(3-methyl-4-phenyl)phenyl]hex-5-enoate (415 mg, 0.70 mmol) in ethanol (20 ml) was hydrogenated over 10% palladium on carbon ( 40 mg) at 3 bar at 20 °C for 2 hours. The mixture was filtered through an Arbocel filter aid and concentrated under reduced pressure. The residue was dissolved in cyclohexane and evaporated (2×), after which methyl (2S,3R)-3-({[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl) was obtained ]amino}carbonyl)propyl]amino}carbonyl)-2-ethoxy-6-(3-fluoro-4-phenoxyphenyl)hexanoate (384 g, 91%) as a colorless foam, which was used in Example 12 without purification.

Rf 0.48 (hexane:ethyl acetate = 1:1).

δH (400 MHz, CDCl 3 ) 1.01 (9H, s); 1.14 (3H, t, J= 7 Hz); 1.49 (3H, d, J = 7 Hz); 1.65 (3H, m); 1.80 (1H, m); 2.24 (3H, s); 2.60 (3H, m); 3.40 (1H, dt, J=16 and 8 Hz); 3.62 (1H, m); 3.70 (3H, s); 4.01 (1H, d, J= 5 Hz); 4.15 (1H, d, J= 9 Hz); 5.13 (1H, pentet, J= 7 Hz); 6.41 (1H, no d); 6.92 (1H, d, J= 9 Hz); 7.00 (1H, d, J= 7 Hz); 7.03 (1H, s); 7.11 (1H, d, J= 7 Hz); 7.29 (7H, complex); 7.40 (3H, m).

LRMS (thermospray) m/z = 602 (MH+).

Preparation 19A

tert-butyl (3R)-3-({[(1S)-2,2-dimethyl-1-({[(1S)-2-methoxy-1-phenylethyl]amino)carbonyl)propyl]amino}carbonyl)- 6-[3-methyl-(4-phenyl)phenyl]hexanoate

[image]

a) N-(dimethylpropyl)-N'-ethylcarbodiimide (2.298 g, 12 mmol) was added to a stirred mixture of (2R)-2-[2-(tert-butoxy)-2-oxoethyl]pent-4-enoic acid (2 .13 g, 10 mmol), L-tert-leucine benzyl ester (2.834 g, 11 mmol) (N. Moss et al.: "J. Med. Chem.", 39, 2178, (1996)), N- methylmorpholine (2.40 ml, 22 mmol) and 1-hydroxybenzotriazole hydrate (1.836 g, 12 mmol) in anhydrous dichloromethane (50 ml) under nitrogen at 0 °C. The mixture was allowed to warm slightly to room temperature in a cooling bath. After 20 hours, the mixture was poured into ethyl acetate (250 ml) and washed with 5% aqueous citric acid (2 x 100 ml), saturated aqueous sodium bicarbonate (2 x 70 ml), and brine (70 ml). , and dried (MgSO4) and concentrated under reduced pressure. The residual oil crystallized after the addition of pentane. The solid was filtered off and dried, after which tert-butyl (3R)-3-({[(1S)-2,2-dimethyl-1-(benzyloxycarbonyl)propyl]amino}carbonyl)hex-5-enoate ( 4.068 g, 97%) as a colorless solid.

T.t. 55-57 °C.

Rf 0.37 (hexane:ethyl acetate = 10:1).

δH (300 MHz, CDCl 3 ) 0.95 (9H, s); 1.42 (9H, s); 2.15 (1H, dt, 16 and 6.5 Hz); 2.39 (2H, m); 2.59 (1H, d, J=10 Hz); 2.68 (2H, m); 4.50 (1H, d, J=10 Hz); 5.00 (1H, d, J=10 Hz); 5.04 (1H, d, J=17 Hz); 5.13 (1H, d, J=12 Hz); 5.19 (1H, d, J=12 Hz); 5.71 (1H, m); 6.35 (1H, no d); 7.34 (5H, complex).

LRMS (thermospray) m/z = 418 (MH+).

b) Using the procedure for making preparation 3a, tert-butyl (3R)-3-({[(1S)-2,2-dimethyl-1-(benzyloxycarbonyl)propyl]amino}carbonyl)hex-5-enoate (2.085 g, 5.0 mmol) was reacted with palladium-catalyzed 3-methyl-4-phenylbromobenzene (1.855 g, 7.5 mmol) using N-ethylmorpholine as base at reflux in acetonitrile for 16.5 hours. This was followed by treatment as before, followed by purification by repeated flash chromatography (first column hexane:ether = 4:1; second column toluene:ether = 20:1), and mainly (3R,5E)-3-({[( 1S)-2,2-dimethyl-1-(benzyloxycarbonyl)propyl]amino)carbonyl)-6-[3-methyl-(4-phenyl)phenyl]hex-5-enoate as a pale yellow resin (2.205 g, 75 %). 1H NMR suggested that two alkene isomers, (5Z) and (4E), were also present. The alkene mixture was carried to the next step (see c, below).

δH (400 MHz, CDCl 3 ) 0.95 (9H, s); 1.42 (9H, s); 2.26 (3H, s); 2.35 (1H, m); 2.45 (1H, dd, 3 and 17 Hz); 2.54 (1H, dt, J= 15 and 6.5); 2.69 (1H, dd, J= 9 and 17 Hz); 2.77 (1H, m); 4.52 (1H, d, J=10 Hz); 4.97 (1H, d, J=12 Hz); 5.06 (1H, d, J=12 Hz); 6.16 (1H, dt, J=18 and 7 Hz); 6.42 (2H, m); 7.29 (13H, complex).

c) Solution of tert-butyl (3R,5E)-3-({[(1S)-2,2-1-(benzyloxycarbonyl)propyl]amino}carbonyl)-6-[3-methyl-(4-phenyl)phenyl ]hex-5-enoate (2.205 g, 3.78 mmol) in ethanol:water = 10:1 (44 ml) was hydrogenated over 10% palladium on carbon at 3 bar at room temperature for 6 hours. The mixture was filtered through an Arbocel filter aid, washing well with ethanol. The filtrate was concentrated under reduced pressure. The residue was dissolved in toluene and evaporated (3×), dissolved in ether and evaporated (2×), and finally dissolved in ether and precipitated as an oil after addition of hexane. Removal of the solvent under reduced pressure gave tert-butyl (3R)-3-({[(1S)-2,2-1-dimethyl-1-(carboxy)propyl]amino}carbonyl)-6-[3-methyl- (4-Phenyl)phenyl]hexanoate (1.817 g, 97%) as a colorless foam.

T.t. 51-56 °C.

Rf 0.38 (hexane:ether:acetic acid = 50:50:1).

δH (400 MHz, DMSO-d6) 0.94 (9H, s); 1.37 (9H, s, and 1H, m, overlap); 1.50 (3H, m); 2.17 (3H, s); 2.23 (1H, m); 2.39 (1H, dd, J=10 and 15 Hz); 2.53 (2H, m); 2.86 (1H, m); 4.10 (1H, d, J=10 Hz); 7.05 (3H, m); 7.29 (3H, m); 7.40 (2H, t, J= 8 Hz); 7.90 (1H, no. d).

LRMS (thermospray) m/z = 513 (MNH4+).

d) 7-azabenzotriazol-1-yloxytris(pyrrolidino)phosphonium hexafluorophosphate (522 mg, 1.0 mmol) was added to a stirred solution of tert-butyl (3R)-3-({[(1S)-2,2-dimethyl-1 -(carboxy)propyl]amino)carbonyl)-6-[3-methyl-(4-phenyl)phenyl] hexoate (496 mg, 1.0 mmol), ((1S)-2-methoxy-1-phenylethylamine (151 mg, 1.0 mmol) ("J. Amer. Chem. Soc.", 117, 10885, (1995)) and collidine (267 µl, 2.0 mmol) in anhydrous dichloromethane (5 ml) under nitrogen at 0 ° C. After 1 hour, the solution was stirred for another 2.75 hours at 20 ° C. The mixture was poured into ethyl acetate (70 ml) and washed successively with 5% aqueous citric acid solution (2 x 50 ml), sat. aqueous sodium bicarbonate solution (2 x 50 ml), and salt (50 ml). The organic solution was dried (MgSO4) and concentrated under reduced pressure. The residue was purified by flash chromatography (eluting with hexane:ethyl acetate = 3:1) and tert-butyl (3R)-3-({[(1S)-2,2-dimethyl-1-({[(1S)-2-methoxy-1-phenylethyl]amino}carbonyl]propyl]amino}carbonyl was obtained )-6-[3-methyl-(4-phenyl)phenyl]hexanoate (537 mg, 8 5 %) in the form of colorless foam.

Rf 0.16 (hexane.ethyl acetate = 4:1).

δH (400 MHz, CDCl 3 ) 1.02 (9H, s); 1.40 (9H, s, and 1H, m, overlap); 1.55 (2H, m); 1.68 (1H, m); 2.23 (3H, s); 2.32 (1H, m); 2.55 (4H, m); 3.33 (3H, s); 3.62 (2H, d, J = 5 Hz); 4.28 (1H, d, J = 9 Hz); 5.12 (1H, dt, J= 8 and 5 Hz); 6.35 (1H, no d); 6.44 (1H, no d); 6.96 (1H, d, J= 8 Hz); 7.00 (1H, s); 7.10 (1H, d, J= 8 Hz); 7.26 (8H, complex); 7.39 (2H, m).

LRMS (thermospray) m/z = 630 (MH+).

FTIR νmax (KBr disc) 3320; 2930; 1729; 1643; 1543; 1370; 1157; and 700 cm-1.

Preparation 19B

tert-butyl (3R)-3-({[(1S)-2,2-dimethyl-1-({[(1S)-2-methoxy-1-phenylethyl]amino}carbonyl)propyl]amino}carbonyl)- 6-[3-methyl-(4-phenyl)phenyl]hexanoate

[image]

a) Alternative synthesis of 3-methyl-4-phenylbromobenzene was carried out as follows:

A mixture of 5-bromo-2-iodo-toluene (5.01 g, 16.9 mmol), phenylboronic acid (2.26 g, 18.5 mmol), palladium acetate (190 mg, 0.85 mmol), triphenylphosphine ( 440 mg, 1.68 mmol) and a 2M aqueous solution of sodium carbonate (25 ml) in acetone (60 ml) was degassed and heated at reflux under nitrogen for 18 hours. The mixture was cooled and partitioned between ether (200 ml) and water (100 ml). The organic layer was washed with salt (100 ml), dried (MgSO4) and concentrated under reduced pressure to give a yellow oil. Purification by flash chromatography (eluting with hexane) gave 3-methyl-4-phenylbromobenzene (3.298 g, 79 %).

b) A solution of 3-methyl-4-phenylbromobenzene (12.36 g, 50.0 mmol) in anhydrous tetrahydrofuran (30 ml) was added dropwise at reflux to a suspension of magnesium shavings (1.33 g, 50 mmol). in anhydrous tetrahydrofuran (23 ml) containing iodine crystals under nitrogen with mechanical stirring. After the addition was complete, the mixture was diluted with anhydrous tetrahydrofuran (55 mL) and heating was continued for another 1 hour. The arylmagnesium bromide solution was allowed to cool to room temperature and then added dropwise via syringe to a solution of glutinous anhydride (6.27 g, 55 mmol) in anhydrous tetrahydrofuran (125 mL) under nitrogen at -40 °C. After 90 minutes at -40 °C, the mixture was allowed to warm to 0 °C, and hydrochloric acid (1M, 250 ml) was added. It was then extracted with ether (500 ml), washed with brine (200 ml), dried (MgSO4) and concentrated under reduced pressure. The residue was purified by flash chromatography (eluting with dichloromethane:methanol = 95:5) and 5-(3-methyl-4-phenyl)phenyl-5-oxopentanoic acid was obtained as a light brown oil (8.6 g, 60 %) , which then crystallized.

T.t. 91-94 °C.

Rf 0.26 (dichloromethane: methanol = 95.5).

δH (400 MHz, CDCl 3 ) 2.11 (2H, pentet, J = 7 Hz); 2.33 (3H, s); 2.50 (2H, t, J= 7 Hz); 3.11 (2H, t, J = 7 Hz); 7.31 (3H, m); 7.42 (3H, m); 7.84 (1H, d, J= 8 Hz); 7.90 (1H, s).

LRMS (thermospray) 283 (MH+).

Found: C, 76.49; H, 6.48.

C18H18O3 required: C, 76.57; H, 6.43%.

c) Triethylsilane (1.4 ml, 8.75 mmol) was added dropwise over 2 minutes to a stirred solution of 5-(3-methyl-4-phenyl)phenyl-5-oxopentanoic acid (1.0 g, 3 .5 mmol) in trifluoroacetic acid (5 ml) under nitrogen at 0 °C. The mixture was then allowed to warm to room temperature and stirred for 2 hours. The mixture was then poured into water (20 ml) and extracted with dichloromethane (3 x 15 ml). The combined organic solutions were washed with salt, dried (MgSO4) and concentrated under reduced pressure. The mixture was purified by flash chromatography (eluting with dichloromethane:methanol = 95:5) and a 3:1 mixture of [5-(3-methyl-4-phenyl)phenyl]pentanoic acid and triethylsilanol (1.1 g) was obtained. This material was then dissolved in hexane (5 mL) and sodium hydrogen carbonate (290 mg, 3.5 mmol) was added. The mixture was stirred overnight at room temperature, and then concentrated under reduced pressure. The solid residue was triturated with ethyl acetate and filtered to give sodium [5-(3-methyl-4-phenyl)phenyl]pentanoate (820 mg, 80%). Treatment of this material with hydrochloric acid and solvent extraction gave the free acid as a viscous, colorless resin.

δH (400 MHz, CDCl 3 ) 1.74 (4H, m); 2.27 (3H, s); 2.43 (2H, m); 2.65 (2H, m); 7.06 (1H, d, J= 8 Hz); 7.11 (1H, s); 7.16 (1H, d, J= 8 Hz); 7.33 (3H, m); 7.42 (2H, m).

LRMS (thermospray) m/z = 286 (MNH4+).

Found: C, 80.40; H, 7.59.

C18H20O2 required: C, 80.56; H, 7.51%.

d) Oxalyl chloride (3.1 ml, 35.5 mmol) was added dropwise to a stirred solution of [5-(3-methyl-4-phenyl)phenyl]pentanoic acid (6.80 g, 25.3 mmol) in anhydrous dichloromethane (60 ml) under nitrogen at -10 °C. Dimethylformamide (2 drops) was added, and after 10 minutes the mixture was allowed to warm to room temperature. After 5 hours, the solution was concentrated under reduced pressure, and the residue was dissolved in anhydrous toluene and concentrated under reduced pressure (2×). The residue was dissolved in hexane (150 ml), allowed to stand for 17 hours, and then filtered through an Arbocel filter aid, washing the filtrate with more hexane. The filtrate was then concentrated under reduced pressure to give 5-[(3-methyl-4-phenyl)phenyl]pentanoyl chloride (7.1 g, 97 %).

δH (400 MHz, CDCl 3 ) 1.77 (4H, m); 2.26 (3H, s); 2.63 (2H, t, J = 6.5 Hz); 2.95 (2H, t, J= 6.5 Hz); 7.06 (1H, d, J= 8 Hz); 7.10 (1H, s); 7.36 (5H, m).

A solution of n-butyllithium (9.92 mL, 2.5 M in hexanes, 24.8 mmol) was added dropwise over 15 min to a solution of 4-(S)-benzyloxazolidin-2-one (4.39 g, 24.8 mmol) in anhydrous tetrahydrofuran (70 ml) under nitrogen at -70 °C. The mixture was allowed to warm to -50 °C for 30 min and then cooled again to -70 °C. A solution of [5-(3-methyl-4-phenyl)phenyl]pentanoyl chloride (7.1 g, 24.8 mmol) in anhydrous tetrahydrofuran (10 mL) was added dropwise over 15 min. After 1 hour, the mixture was allowed to warm to 0 °C, after which 20% aqueous ammonium chloride (75 ml) was added rapidly. After stirring for 15 minutes, the mixture was extracted with ethyl acetate (150 ml), and the organic phase was washed with water (3 x 250 ml), dried (MgSO4) and concentrated under reduced pressure. The residue was purified by flash chromatography (gradient elution with pentane:ethyl acetate = 20:1 to 2:1) and 4-(S)-benzyl-3-{[5-(3-methyl-4-phenyl)phenyl] was obtained. pentanoyl}oxazolidin-2-one (9.66 g, 91%) as a colorless oil.

Rf 0.4 (pentane:ethyl acetate = 3:1)

δH (300 MHz, CDCl 3 ) 1.79 (4H, m); 2.28 (3H, s); 2.70 (2H, m); 2.79 (1H, dd, J=10 and 13 Hz); 3.01 (2H, m); 3.32 (1H, dd, J= 3 and 13 Hz); 4.19 (2H, m); 4.70 (1H, m); 7.06-7.43 (13H, complex).

LRMS (thermospray) m/z = 445 (MNH4+).

FTIR (KBr disk) 2930; 1784; 1700; 1387; 1350; 1211; 702 cm-1.

Found: C, 75.43; H, 6.65; N, 3.08.

C28H29NO3 • 0.3 CH2Cl2 requirements: C, 75.03; H, 6.59; N, 3.09%.

e) A solution of sodium hexamethyldisilazide (31.3 ml, 1M in tetrahydrofuran, 31.3 mmol) was added dropwise over 30 minutes to a stirred solution of 4-(S)-benzyl-3-{[5-(3-methyl -4-phenyl)phenyl]pentanoyl}oxazolidin-2-one (13.39 g, 31.3 mmol) in anhydrous tetrahydrofuran (100 ml) at -75 °C. After 1 h, a solution of tert-butyl bromoacetate (4.95 mL, 33.5 mmol) in anhydrous tetrahydrofuran (10 mL) was added dropwise over 20 min, maintaining the temperature below -70 °C. The mixture was stirred at this temperature for 2 hours and then allowed to warm to -50 °C, when 20% ammonium chloride solution (150 ml) was added with rapid stirring. The mixture was allowed to warm to 10 °C, after which it was poured into a mixture of ethyl acetate (400 ml) and water (200 ml). The organic phase was removed, washed with water (3 x 250 ml), dried (MgSO 4 ) and concentrated under reduced pressure. The residue was purified by flash chromatography (gradient elution with pentane:ether = 20:1 to 1:1) and tert-butyl (3R)-3-[(4-(S)-4-benthyl-2-oxo-1, 3-Oxazolidin-3-yl]-6-(3-methyl-4-phenyl)phenyl)hexanoate as a colorless oil (10.4 g, 61 %).

Rf 0.6 (pentane:ether = 1:1).

δH (300 MHz, CDCl 3 ) 1.45 (9H, s); 1.58 (1H, m); 1.74 (3H, m); 2.27 (3H, s); 2.51 (1H, dd, J= 5 and 18 Hz); 2.65 (2H, m); 2.80 (2H, m); 3.38 (1H, dd, J= 3 and 15 Hz); 4.16 (2H, d, J = 5 Hz); 4.25 (1H, m); 4.68 (1H, m); 7.04 (1H, d, J= 8 Hz); 7.08 (1H, s); 7.14 (1H, d, J= 8 Hz); 7.35 (10H, complex).

LRMS (thermospray) = m/z =542 (MH + ), 560 (MNH 4 + ).

Found: C, 74.55; H, 7.40; N, 2.44.

C34H39NO5 • 0.2 EtOAc requirements: C, 74.60; H, 7.40; N, 2.47%.

FTIR νmax (KBr disk) 2980; 2930; 1780; 1725; 1700; 1388; 1350; 1157; 768; 702 cm-1.

f) A 30% aqueous solution of hydrogen peroxide (12.75 ml, 114 mmol) was added dropwise to the tert-butyl (3R)-3-[(4-(S)-4-benzyl-2- oxo-1,3-oxazolidin-3-yl)carbonyl]-6-(3-methyl-4-phenyl)phenyl)hexanoate (10.3 g, 19.0 mmol) in tetrahydrofuran:water (3:1, 400 ml) at 0 °C. Lithium hydroxide monohydrate (1.395 g, 38 mmol) was then added in one portion. The mixture was stirred for 2 hours at 0 °C and 1 hour at 20 °C. The reaction mixture was cooled again to 0 °C and a solution of sodium sulfite (15.56 g, 123.5 mmol) in water (80 ml) was added dropwise over 15 minutes. The mixture was stirred vigorously at 0 °C for 2.5 hours, then 2M hydrochloric acid (about 8 ml) was added to adjust the pH to 6. The mixture was concentrated under reduced pressure to half the volume, acidified to pH 2 by the addition of 2M hydrochloric acid. and extracted with dichloromethane (400 ml, 2 × 200 ml). The combined extracts were dried (MgSO4), concentrated under reduced pressure, and the residue was purified by flash chromatography (gradient elution with pentane:ether = 15:1 to 1:4) and tert-butyl (3R)-3-(carboxy) was obtained. -6-(3-methyl-4-phenyl)phenyl)hexanoate (7.05 g, 97%) as a colorless oil.

Rf 0.5 (pentane:ether:acetic acid = 30:70:1).

δH (300 MHz, CDCl 3 ) 1.45 (9H, s); 1.63 (1H, m); 1.77 (4H, m); 2.26 (3H, m); 2.42 (dd, J= 5 and 17 Hz); 2.65 (3H, m); 2.87 (1H, m); 7.06 (2H, m); 7.15 (1H, d, J= 8 Hz); 7.34 (5H, m).

LRMS (thermospray) m/z = 400 (MNH4+).

[α]D +12.9 ° (c = 0.716, methanol, 25 °C).

FTIR νmax (film) 2980; 2930; 1730; 1705; 1485; 1368; 1157; 764; 702 cm-1.

Found: C, 74.96; H, 8.06.

C24H30O4 • 0.1 H2O requirements: C, 75.01; H, 7.92%.

g) N-(dimethylaminopropyl)-N'-ethylcarbodiimide hydrocloprid (3.50 g, 18.26 mmol) was added to the stirred mixture of (2S)-amino-3,3-dimethyl-N-[(1S)-2-methoxy -1-phenylethyl]butanamide hydrochloride (preparation 1a) (6.78 g, 17.43 mmol - contains 1 mol equiv. of dioxane), tert-butyl (3R)-3-(carboxy)-6-(3-methyl- 4-phenyl)phenyl)hexanoate (6.35 g, 16.6 mmol), 1-hydroxy-1,2,3-benzotriazole hydrate (2.80 g, 20.75 mmol) and diisopropylethylamine (5.9 ml, 34.03 mmol) in anhydrous dichloromethane (82 ml) under nitrogen at 4 °C. After 2 hours, the mixture was allowed to warm to room temperature. After 17 hours at this temperature, the mixture was poured into ethyl acetate (600 ml) and washed successively with 5% aqueous citric acid solution (2 x 250 ml), saturated aqueous sodium bicarbonate solution (2 x 250 ml), and with salt (200 ml), and dried (MgSO4) and concentrated under reduced pressure. The residue was re-dissolved in ether and evaporated, after which tert-butyl (3R)-3-({[(1S)-2,2-dimethyl-1-({[(1S)-2-methoxy-1) was obtained -phenylethyl]amino}carbonyl)propyl]amino}carbonyl)-6-[3-methyl-(4-phenyl)phenyl]hexanoate (10.35 g, 99%) as a colorless foam.

Found: C, 73.99; H, 8.31; N, 4.51.

C39H52N2O5 • 0.04 EtOAC required: C, 74.38; H, 8.34; N, 4.43%.

Preparation 20

4-iodo-1-(3-methoxyphenyl)-2-methylbenzene

[image]

a) A solution of 2-bromo-5-nitrotoluene (40.0 g, 185 mmol) in 1,2-dimethoxymethane (600 ml) was added to tetrakis-(triphenylphosphine)palladium (0) (9.77 g, 9.25 mmol ) under nitrogen at room temperature. A solution of 3-methoxyphenylboronic acid (31.5 g, 207 mmol) in ethanol (150 ml) was added. Finally, a 2M aqueous solution of sodium carbonate (800 ml) was added, and the mixture was heated under reflux with strong mechanical stirring for 18 hours. After cooling, the mixture was partitioned between ethyl acetate and water, and the organic phase was dried (Na2SO4) and concentrated under reduced pressure. Purification by column chromatography over silica gel eluting with hexane:ethyl acetate = 98:2 to 95:5 gave 1-(3-methoxyphenyl)-2-methyl-4-nitrobenzene (40.55 g, 90 %).

Rf 0.22 (hexane:ethyl acetate = 95:5).

δH (400 MHz, CDCl 3 ) 2.38 (3H, s); 3.85 (3H, s); 6.83 (1H, s); 6.88 (1H, d, J = 8 Hz); 6.96 (1H, d, J= 8 Hz); 7.38 (2H, m); 8.02 (1H, d, J= 8 Hz); 8.14 (1H, s).

LRMS (thermospray) m/z = 261 (MNH4+).

b) 1-(3-methoxyphenyl)-2-methyl-4-nitrobenzene (23.4 g, 96 mmol) was dissolved in a mixture of ethanol and ethyl acetate (3:1, 470 ml) and hydrogenated over 10% palladium on carbon (2.3 g) at 3 bar at 20 °C for 2 hours. The mixture was filtered through an Arbocel filter aid, washing with ethyl acetate. The filtrate was concentrated under reduced pressure to give 4-amino-1-(3-methoxyphenyl)-2-methylbenzene (20.8 g, 100%) as a purple-brown oil.

Rf 0.06 (hexane:ethyl acetate = 90:10).

δH (400 MHz, CDCl 3 ) 2.22 (3H, s); 3.65 (2H, no s); 3.84 (3H, s); 6.59 (2H, m); 6.87 (3H, m); 7.04 (1H, d, J= 8 Hz); 7.28 (1H, m).

c) A mixed solution of 4-amino-1-(3-methoxyphenyl)-2-methylbenzene (5.0 g, 23.4 mmol) and iodine (2.97 g, 25.8 mmol) in toluene (120 ml) heated was at 50 °C, after which a solution of iso-amyl nitrite (3.46 ml, 25.8 mmol) in toluene (50 ml) was added. The mixture was heated to 90 °C for 2 hours, after which it was cooled. Toluene was removed under reduced pressure, and the residue was dissolved in dichloromethane and washed with an excess of 5% aqueous sodium metabisulfite to remove iodine. The organic solution was dried (Na2SO4) and concentrated under reduced pressure. The residue was purified by repeated flash chromatography (first column, gradient elution with hexane:ethyl acetate and pure hexane; second column also eluted with hexane) to give 4-iodo-1-(3-methoxyphenyl)-2-methylbenzene as a pale orange oil. (5.63 g, 74 %).

Rf 0.49 (hexane:ethyl acetate = 90:10).

δH (400 MHz, CDCl 3 ) 2.23 (3H, s); 3.84 (3H, s); 6.81 (1H, s); 6.86 (1H, d, J= 8 Hz); 6.90 (1H, d, J= 8 Hz); 6.95 (1H, d, J= 8 Hz); 7.32 (1H, t, J= 8 Hz); 7.55 (1H, d, J= 8 Hz); 7.47 (1H, s).

Preparation 21

tert-butyl (3R)-3-({[(1S)-2,2-dimethyl-({[(1S)-2-methoxy-1-phenylethyl]amino}carbonyl)propyl]amino}carbonyl)-6- (3'-Methoxy-2-methylbiphen-4-yl)hexanoate

[image]

a) Using the procedure for preparing preparation 3a, tert-butyl (3R)-3-({[(1S)-2,2-dimethyl-1-(benzyloxycarbonyl)propyl]amino}carbonyl)hex-5-enoate (2.085 g, 5.0 mmol) was reacted with 4-iodo-1-(3-methoxyphenyl)-2-methylbenzene (preparation 20) (889 mg, 2.13 mmol) catalyzed by palladium, using N-ethylmorpholine as a base, at reflux in acetonitrile for 14.5 hours. Processing as before, followed by purification by flash chromatography (hexane:ethyl acetate = 10:1) gave mainly tert-butyl (3R,5E)-3-({[(1S)-2,2-dimethyl-1-(benzyloxycarbonyl) )propyl]amino}carbonyl)-6-(3'-methoxy-2-methylbiphen-4-yl)-hex-5-enoate as a pale yellow resin (1.043 g, 80 %). 1H NMR suggested that two alkene isomers were also present. The alkene mixture is transferred to the next step (see b, below).

δH (400 MHz, CDCl 3 ) 0.95 (9H, s); 1.43 (9H, s); 2.26 (3H, s); 2.36 (1H, m); 2.45 (1H, dd, 3 and 17 Hz); 2.55 (1H, dt, J=14 and 7); 2.68 (1H, dd, J= 9 and 17 Hz); 2.77 (1H, m); 3.80 (3H, s); 4.50 (1H, d, J=10 Hz); 4.97 (1H, d, J=12 Hz); 5.07 (1H, d, J=12 Hz); 6.16 (1H, dt, J=18 and 7 Hz); 6.40 (2H, m); 6.85 (3H, m); 7.30 (9H, complex).

LRMS (thermospray) m/z = 614 (MH+).

b) According to the procedure for making preparation 19c, tert-butyl (3R,5E)-3-({[(1S)-2,2-dimethyl-1-(benzyloxycarbonyl)propyl]amino}carbonyl)-6-(3' -Methoxy-2-methylbiphen-4-yl]hex-5-enoate (1.040 g, 1.69 mmol) was hydrogenated over 10% palladium on carbon to give tert-butyl (3R)-3-({[ (1S)-2,2-dimethyl-1-(carboxy)propyl]amino}carbonyl)-6-(3'-methoxy-2-methylbiphen-4-yl)hexanoate (664 g, 75%) as a colorless foam .

Rf 0.33 (hexane:ether:acetic acid = 50:50:1).

δH (400 MHz, CDCl 3 ) 1.02 (9H, s); 1.42 (9H, s); 1.47 (1H, m); 1.66 (2H, m); 1.77 (1H, m); 2.25 (3H, s); 2.37 (1H, m, ); 2.63 (3H, m); 3.83 (3H, s); 4.48 (1H, d, J=10 Hz); 6.43 (1H, no d); 6.87 (3H, m); 7.03 (2H, m); 7.13 (1H, d, J= 8 Hz); 7.28 (1H,m).

LRMS (thermospray) m/z = 526 (MH+).

c) According to the procedure for making preparation 19d, tert-butyl (3R)-3-({[(1S)-2,2-dimethyl-1-(carboxy)propyl]amino}carbonyl)-6-(3'-methoxy -2-Methylbiphen-4-yl)hexanoate (660 mg, 1260 mmol) was reacted with (1S)-2-methoxy-1-phenylethylamine (190 mg, 1.260 mmol). Processing as above, followed by flash chromatography (elution with hexane:ethyl acetate = 3:1) gave tert-butyl (3R)-({[(1S)-2,2-dimethyl-1-({[(1S) -2-Methoxy-1-phenylethyl]amino}carbonyl)propyl]amino}carbonyl)-6-(3'-methoxy-2-methylbiphen-4-yl)hexanoate (672 mg, 81%) as a colorless foam.

Rf 0.16 (hexane:ethyl acetate = 4:1).

δH (400 MHz, CDCl 3 ) 1.02 (9H, s); 1.40 (9H, s, and 1H, m, overlap); 1.55 (2H, m); 1.68 (1H, m); 2.23 (3H, s); 2.33 (1H, m); 2.56 (4H, m); 3.33 (3H, s); 3.63 (2H, m); 3.82 (3H, s); 4.29 (1H, d, J= 9 Hz); 5.13 (1H, dt, J= 8 and 5 Hz); 6.35 (1H, no d); 6.45 (1H, no d); 6.86 (3H, m); 6.96 (1H, d, J= 8 Hz); 7.00 (1H, s); 7.10 (1H, d, J= 8 Hz); 7.26 (6H, complex).

LRMS (thermospray) m/z = 659 (MH+).

Found: C, 72.15; H, 8.21; N, 4.17.

C40H54N2O6 • 0.5 H2O requirements: C, 71.93; H, 8.30; N, 4.19%.

Preparation 22

(2R)-5-{[4-(4-cyanophenyl)-3-methyl]phenyl}-2-[(4S)-2,2-dimethyl-5-oxo-1,3-dioxolan-4-yl] -N-[(1S)-2,2-dimethyl-1-({[(1R)

-1-phenylethyl]amino}carbonyl)propyl]pentanamide

[image]

a) According to the procedure for making preparation 15a, (2R)-2-[(4S)-2,2-dimethyl-5-oxo-1,3-dioxolan-4-yl]-N-[(1S)-2, 2-Dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]pent-4-enamide (preparation 14) (479 mg, 1.5 mmol) reacts with 1-(4-cyanophenyl) -4-iodo-2-methylbenzene (preparation 23) (479 mg, 1.5 mmol) under palladium catalysis at reflux in acetonitrile for 16 h. After cooling, the mixture was partitioned between ethyl acetate (3 x 30 mL) and saturated aqueous sodium bicarbonate (30 mL). The combined organic solutions were dried (NaSO4) and concentrated under reduced pressure. The residue was purified by flash chromatography (eluting with dichloromethane:methanol = 97.5:2.5) and obtained mainly (2R,4E)-5-{[4-(4-cyanophenyl)-3-methyl]phenyl}-2 -[(4S)-2,2-dimethyl-5-oxo-1,3-dioxolan-4-yl]-N-[(1S)-2,2-dimethyl-1-({[(1R)-1 -phenylethyl]amino}carbonyl)propyl]pent-4-anamide, as a light brown foam (403 mg, 65 %). 1H NMR suggested that two alkene isomers were also present. The alkene mixture was carried to the next step (see b, below).

Rf 0.41 (dichloromethane: methanol = 95:5).

δH (400 MHz, CDCl 3 ) (for (4E) isomer) 1.00 (9H, s); 1.46 (3H, d, J = 7.5 Hz); 1.54 (3H, s); 1.59 (3H, s); 2.20 (3H, s); 2.70 (1H, m); 2.81 (2H, m); 4.20 (1H, d, J= 9 Hz); 4.60 (1H, d, J= 4.5 Hz); 5.04 (1H, pentet, J = 7.5 Hz); 5.82 (1H, d, J = 7.5 Hz); 6.62 (1H, dt, 16 and 7 Hz); 6.50 (1H, d, J=16 Hz); 6.58 (1H, d, J= 9 Hz); 7.10 (1H, d, J= 8 Hz); 7.15-7.30 (7H, m); 7.38 (2H, d, J = 8 Hz); 7.70 (2H, d, J= 8 Hz).

LRMS (thermospray) m/z = 564 (MH+ - acetone); 547.

b) Solution of (2R,4E)-5-{[4-(4-cyanophenyl)-3-methyl]phenyl}-2-[(4S)-2,2-dimethyl-5-oxo-1,3-dioxolane -4-yl]-N-[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]pent-4-anamide (403 mg, 0.65 mmol) in ethanol (70 ml) was hydrogenated over 10% palladium on carbon (40 mg) at 3 bar at 20 °C for 5.5 hours. The mixture was filtered through an Arbocel filter aid and concentrated under reduced pressure to give (2R)-5-{[4-(4-cyanophenyl)-3-methyl]phenyl}-2-[(4S)-2,2-dimethyl -5-oxo-1,3-dioxolan-4-yl]-N-[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]pentanamide ( 384 mg, 61 %) as a colorless solid, which was used in example 17 without purification.

δH (400 MHz, CDCl 3 ) 1.00 (9H, s); 1.48 (3H, d, J = 7 Hz); 1.50 (3H, s); 1.55 (3H, s); 1.62 (2H, m); 1.68 (2H, m); 2.20 (3H, s); 2.60 (3H, m); 4.18 (1H, d, J= 9.5 Hz); 4.45 (1H, d, J= 6 Hz); 5.08 (1H, pentet, J= 7 Hz); 5.86 (1H, d, J = 7 Hz); 6.50 (1H, d, J= 9.5 Hz); 7.00-7.10 (3H, m); 7.15-7.30 (5H, m); 7.40 (2H, d, J = 7.5 Hz); 7.70 (2H, d, J = 7.5 Hz).

LRMS (thermospray) m/z = 624 (MH+); 641 (MNH4+).

Preparation 23

1-(4-Cyanophenyl)-4-iodo-2-methylbenzene

[image]

a) A mixture of 2-bromo-5-nitrotoluene (7.05 g, 32.47 mmol), 4-cyanophenylboronic acid ("Tet. Lett.", 34, 8237, (1993)) (6.0 g, 40 .83 mmol), cesium fluoride (11.2 g, 72.53 mmol) and tri-(2-methylphenyl)phosphine (1.0 g, 3.27 mmol) in 1,2-dimethyloxyethane (240 ml) was mixed in a nitrogen atmosphere at room temperature for 40 minutes. Tris(dibenzylideneacetone)dipalladium (1.50 g, 1.63 mmol) was added and stirred for another 10 minutes at room temperature and then for another 4 hours at 80 °C. After cooling, the mixture was partitioned between ethyl acetate and saturated aqueous sodium bicarbonate. The organic phase was separated, washed with water, dried (Na2SO4) and concentrated under reduced pressure, after which crude 1-(4-cyanophenyl)-2-methyl-4-nitrobenzene (9.78 g) was obtained, which was further used without purification.

Rf 0.89 (dichloromethane).

δH (300 MHz, CDCl 3 ) 2.35 (3H, s); 7.35 (1H, d, J= 8 Hz); 7.45 (2H, d, J= 8 Hz); 7.68 (2H, d, J=8 Hz); 8.10 (1H, d, J=8 Hz); 8.18 (1H, s).

b) 1-(4-cyanophenyl)-2-methyl-4-nitrobenzene (9.7 g, 40.76 mmol) was dissolved in a mixture of methanol and dichloromethane (1:1, 400 ml) and hydrogenated over 10% palladium on carbon (970 mg) at 3 bar at 20 °C for 18 hours. The mixture was then filtered through an Arbocel filter aid and concentrated under reduced pressure. The residue was purified by flash chromatography (gradient elution with hexane:ethyl acetate = 20:1 to 1:1) to give 4-amino-1-(4-cyanophenyl)-2-methylbenzene (2.5 g, 30 %) in see brown oil.

Rf 0.20 (hexane:ethyl acetate = 3:1).

δH (400 MHz, CDCl 3 ) 2.20 (3H, s); 3.75 (2H, no s); 6.60 (2H, m); 7.00 (1H, d, J= 8 Hz); 7.40 (2H, d, J= 8 Hz); 7.66 (2H, d, J = 8 Hz).

LRMS (thermospray) = 226 (MNH4+).

c) A suspension of 4-amino-1-(4-cyanophenyl)-2-methylbenzene (250 mg, 1.2 mmol) in concentrated hydrochloric acid (1.3 ml) and water (1.6 ml) at 0 °C, was treated with a solution of sodium nitrite (183 mg, 1.65 mmol) in water (1 ml) for 10 minutes. The mixture was stirred for 20 minutes, then a solution of potassium iodide (840 mg, 5.06 mmol) in water (2 ml) was added over 10 minutes. The resulting mixture was heated to 90 °C for 2 hours and then cooled. The residue was extracted with dichloromethane (3 × 30 ml), and the combined organic extracts were washed with 5% aqueous sodium metabisulfite (4 × 30 ml) to remove iodine. The organic solution was dried (Na2SO4) and concentrated under reduced pressure. The residue was purified by flash chromatography (gradient elution with hexane:ethyl acetate 20:1 to 10:1) to give 1-(4-cyanophenyl)-4-iodo-2-methylbenzene as a white solid (263 mg, 69%) ).

Rf 0.62 (hexane:ethyl acetate = 4:1).

δH (300 MHz, CDCl 3 ) 2.20 (3H, s); 6.92 (1H, d, J= 8 Hz); 7.40 (2H, d, J= 8 Hz); 7.60 (1H, d, J= 8 Hz); 7.68 (1H, s); 7.74 (2H, d, J= 8 Hz).

LRMS (thermospray) m/z = 319 (MH+).

Preparation 24

(2R)-5-{[4-cyanophenyl)-3-methyl]phenyl}-2-[(4S)-2,2-dimethyl-5-oxo-1,3-dioxolan-4-yl]-N- [(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]pentanamide

[image]

a) Palladium acetate (12.2 mg, 0.050 mmol) was added to a stirred solution of (2R)-2-[(4S)-2,2-dimethyl-5-oxo-1,3-dioxolan-4-yl]-N -[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]pent-4-enamide (preparation 14) (430 mg, 1.0 mmol), 1-(3-cyanophenyl)-4-iodo-2-methylbenzene (preparation 25) (457 mg, 1.50 mmol), tri-(2-methylphenyl)phosphine (30 mg, 0.10 mmol) and N-ethylmorpholine (190 µl, 1.5 mmol) in anhydrous acetonitrile (3 ml) under nitrogen. The mixture was flushed with nitrogen and then heated at reflux for 14 hours. After cooling, the mixture was dissolved in ethyl acetate (100 ml), washed with water (100 ml), 0.5 M aqueous sodium dihydrogen phosphate (50 ml) and saturated aqueous sodium bicarbonate (50 ml). The organic solution was dried (Na2SO4) and concentrated under reduced pressure. The residue was purified by flash chromatography (gradient elution with hexane:ethyl acetate = 3:1 to 1:1), and obtained mainly (2R,4E)-5-{[4-(3-cyanophenyl)-3-methyl]phenyl }-2-[(4S)-2,2-dimethyl-5-oxo-1,3-dioxolan-4-yl]-N-[(1S)-2,2-dimethyl-1-({[(1R )-1-phenylethyl]amino}carbonyl)propyl]pent-4-enamide, as a brown foam (385 mg, 62 %). 1H NMR suggested that two alkene isomers were also present. The alkene mixture was carried to the next step (see b, below).

Rf 0.27 (dichloromethane:ethyl acetate = 10:1).

δH (400 MHZ, CDCl 3 ) (for (4E) isomer) 1.01 (9H, s); 1.50 (3H, d, J= 7 Hz); 1.55 (3H, s); 1.60 (3H, s); 2.20 (3H, s); 2.70 (1H, m); 2.82 (2H, m); 4.18 (1H, d, J= 8.5 Hz); 4.58 (1H, d, J= 5 Hz); 5.04 (1H, pentet, J= 7 Hz); 5.84 (1H, d, J = 7 Hz); 6.16 (1H, dt, 16 and 7 Hz); 6.50 (1H, d, J=16 Hz); 6.58 (1H, d, J = 8.5 Hz); 7.05 (1H, d, J= 8 Hz); 7.10-7.40 (8H, complex); 7.54 (2H, m); 7.62 (1H, m).

LRMS (thermospray) m/z = 639 (MNH4+).

Found: C, 72.88; H, 7.05; N, 6.63.

C38H43N3O5 • 0.25 H2O requirements: C, 72.88; H, 7.00; N, 6.71%.

b) Solution of (2R,4E)-5-{[4-(3-cyanophenyl)-3-methyl]phenyl}-2-[(4S)-2,2-dimethyl-5-oxo-1,3-dioxolane -4-yl]-N-[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]pent-4-enamide (359 mg, 0.58 mmol) in ethanol (35 ml) was hydrogenated over 10% palladium on carbon (35 mg) at 3 bar at 20 °C for 6 hours. The mixture was filtered through an Arbocel filter aid, concentrated under reduced pressure and azeotripped with ethyl acetate and then with diethyl ether. The residual oil was purified by flash chromatography (gradient elution with hexane:ethyl acetate 5:1 to 1:1), then triturated with pentane and crystallized to give (2R)-5-{[4-(3-cyanophenyl)-3-methyl ]phenyl)]-2-[(4S)-2,2-dimethyl-5-oxo-1,3-dioxolan-4-yl]-N-[(1S)-2,2-dimethyl-1-({ [(1R)-1-phenylethyl]amino}carbonyl)propyl]pentanamide (139 mg, 38%) as a white solid, which was used in Example 19.

Rf 0.40 (hexane:ethyl acetate = 1:1).

δH (400 MHz, CDCl 3 ) 1.02 (9H, s); 1.52 (3H, d, J= 7 Hz); 1.55 (3H, s); 1.58 (3H, s); 1.64 (2H, m); 1.90 (2H, m); 2.20 (3H, s); 2.60 (3H, m); 4.20 (1H, d, J = 9.5 Hz); 4.46 (1H, d, J = 7.5 Hz); 5.08 (1H, pentet, J= 7 Hz); 5.86 (1H, d, J = 7 Hz); 6.50 (1H, d, J= 9.5 Hz); 6.98-7.08 (3H, m); 7.20 (5H, m); 7.50 (2H, m); 7.55 (1H, s); 7.60 (1H, m).

LRMS (thermospray) m/z = 641 (MHN4+).

Found: C, 72.99; H, 7.36; N, 6.75.

C38H45N3O5 required: C, 73.17; H, 7.27; N, 6.74%.

Preparation 25

1-(3-Cyanophenyl)-4-iodo-2-methylbenzene

[image]

a) A solution of 2-bromo-5-nitrotoluene (8.82 g, 40.83 mmol) in 1,2-dimethoxyethane (150 ml) was added to a solution of tetrakis(triphenylphosphine)palladium (0) (2.35 g, 2, 04 mmol) in 1,2-dimethoxyethane (100 ml) under nitrogen at room temperature. A solution of 3-cyanophenylboronic acid ("J. Med. Chem.", 40, 4208, (1997)) (9.0 g, 61.25 mmol) in ethanol (45 ml) was added and stirred for 10 minutes. Finally, a 2M aqueous solution of sodium carbonate (234 ml) was added, and the mixture was heated at reflux for 18 hours. After cooling, the mixture was partitioned between ethyl acetate and saturated aqueous sodium bicarbonate. The organic phase was washed with water, dried (Na2SO4), concentrated under reduced pressure and triturated with diethyl ether to give 1-(3-cyanophenyl)-2-methyl-4-nitrobenzene (8.48 g, 87%) as a brown solid matter.

Rf 0.74 (hexane:ethyl acetate 1:1).

δH (300 MHz, CDCl 3 ) 2.35 (3H, s); 7.38 (1H, d, J= 8 Hz); 7.52-7.68 (3H, m); 7.75 (1H, d, J= 8 Hz); 8.15 (1H, d, J= 8 Hz); 8.20 (1H, s).

LRMS (thermospray) m/z = 239 (MH+).

b) 1-(3-cyanophenyl)-2-methyl-4-nitrobenzene (3.0 g, 12.61 mmol) and stannous chloride dihydrate were suspended in ethanol (30 ml) and heated to 70 °C for 30 minutes. After cooling, the mixture was placed on ice and neutralized by adding sodium bicarbonate. The mixture was then extracted with dichloromethane (3 x 50 ml). The combined organic layers were dried (Na2SO4) and concentrated under reduced pressure to give 4-amino-1-(3-cyanophenyl)-2-methylbenzene (1.8 g, 69%) as a yellow oil.

Rf 0.36 (hexane:ethyl acetate = 2:1).

δH (300 MHz, CDCl 3 ) 2.20 (3H, s); 3.75 (2H, no s); 6.60 (2H, m); 7.00 (1H, d, J= 8 Hz); 7.42-7.60 (4H, m).

c) A suspension of 4-amino-1-(3-cyanophenyl)-2-methylbenzene (2.5 g, 12.0 mmol) in concentrated hydrochloric acid (13 ml) and water (16 ml) at 0 °C was treated with a solution of sodium nitrite (1.83 g, 26.5 mmol) in water (10 ml) for 10 minutes. The mixture was stirred for 20 min, then a solution of potassium iodide (8.40 g, 50.6 mmol) in water (20 mL) was added over 10 min. The resulting mixture was heated to 80 °C for 2 hours, and then cooled. The residue was extracted with dichloromethane, after which the combined organic extracts were washed with an excess of 5% aqueous sodium metabisulfite to remove iodine. The organic solution was dried (Na2SO4) and concentrated under reduced pressure. The residue was purified by flash chromatography (gradient elution with hexane:ethyl acetate 20:2 to 10:1) to give 1-(3-cyanophenyl)-4-iodo-2-methylbenzene as a white solid (1.105 g, 29 %).

Rf 0.67 (hexane:ethyl acetate = 4:1).

δH (300 MHz, CDCl 3 ) 2.20 (3H, s); 6.90 (1H, d, J= 8 Hz); 7.45-7.75 (6H, complex).

LRMS (APCI) m/z = 319 (MH + ).

Preparation 26

(2R)-5-{[4-(3-carbamoylphenyl)-3-methyl]phenyl}-2-[(4S)-2,2-dimethyl-5-oxo-1,3-dioxolan-4-yl] -N-[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]pentanamide

[image]

a) Palladium acetate (10.4 mg, 0.042 mmol) was added to a stirred solution of (2R)-2-[(4S)-2,2-dimethyl-5-oxo-1,3-dioxolan-4-yl]-N -[(1S)-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]pent-4-enamide (preparation 14) (366 mg, 0.85 mmol), 1-(3 -carbamoylphenyl)-4-iodo-2-methylbenzene (preparation 27) (300 mg, 0.89 mmol), tri-(2-methylphenyl)phosphine (27 mg, 0.09 mmol) and N-ethylmorpholine (115 µl, 0.89 mmol) in anhydrous acetonitrile (3 mL) under nitrogen. The mixture was flushed with nitrogen and then heated at reflux for 3 hours. Palladium acetate (10.4 mg, 0.042 mmol) was added and refluxed for 60 min. More portions of palladium acetate (10.4 g, 0.042 mmol) were added at 60 minute intervals, followed by reflux, until the reaction was refluxed for 8 hours. The reaction mixture was cooled, diluted with acetonitrile (10 ml) and filtered through an Arbocel filter aid, washing with ethyl acetate (100 ml). The filtrate was washed with 5% aqueous sodium bicarbonate solution (50 ml), dried (Na2SO4) and concentrated under reduced pressure. The residue was purified by flash chromatography (gradient elution with dichloromethane:ethyl acetate = 10:1 to 1:1) and obtained (2R,4E)-5-{[4-(3-carbamoylphenyl)-3-methyl]phenyl]- 2-[(4S)-2,2-dimethyl-oxo-1,3-dioxolan-4-yl]-N-[(1S)-2,2-dimethyl-1-({[(1R)-1- phenylethyl]amino}carbonyl)propyl]pent-4-enamide, in the form of a yellow foam (175 mg, 32 %).

Rf 0.20 (dichloromethane: ethyl acetate = 1:1).

δH (400 MHz, CDCl 3 ) 1.02 (9H, s); 1.50 (3H, d, J= 7 Hz); 1.54 (3H, s); 1.60 (3H, s); 2.20 (3H, s); 2.70 (1H, m); 2.78 (1H, m); 2.88 (1H, m); 4.22 (1H, d, J= 8.5 Hz); 4.60 (1H, d, J= 5.5 Hz); 5.04 (1H, pentet, J= 7 Hz); 5.94 (2H, no s); 6.00 (1H, d, J= 7 Hz); 6.12 (1H, dt, J= 16 and 7 Hz); 6.48 (1H, d, J=16 Hz); 6.82 (1H, d, J = 8.5 Hz); 7.08 (1H, d, J = 7.5 Hz); 7.14 (1H, m); 7.18-7.36 (5H, complex); 7.40-7.52 (3H, m); 7.60 (1H, s); 7.80 (1H, d, J= 7.5 Hz).

LRMS (thermospray) m/z = 657 (MNH4+).

Found: C, 70.59; H, 7.16; N, 6.42.

C38H45N3O6 • 0.2 EtOAc required: C, 70.89; H, 7.14; N, 6.39%.

b) Solution of (2R,4E)-5-{[4-(3-carbamoylphenyl)-3-methyl]phenyl]-2-[(4S)-2,2-dimethyl-oxo-1,3-dioxolane-4 -yl]-N-[(1S)-2,2-dimethyl-1-({[(1R)-1-phenylethyl]amino}carbonyl)propyl]pent-4-enamide (160 mg, 0.25 mmol) in ethanol (50 ml) was hydrogenated over 10% palladium on carbon (26 mg) at 3 bar at 20 °C for 8 hours. The mixture was filtered through an Arbocel filter aid, concentrated under reduced pressure and azeotroped with ethyl acetate and then with diethyl ether. The residue was purified by flash chromatography (gradient elution with hexane:ethyl acetate 5:1 to 1:3), then azeotroped with diethyl ether to give (2R)-5-{[4-(3-carbamoylphenyl)-3-methylphenyl} -2-[(4S)-2,2-dimethyl-5-oxo-1,3-dioxolan-4-yl]-N-[(1S)-2,2-dimethyl-1-({[(1R) -1-phenylethyl]amino}carbonyl)propyl]pentanamide (82 mg, 51%) as a white solid, which was used in Example 20.

Rf 0.17 (hexane.ethyl acetate = 1:3).

δH (400 MHz, DMSO) 0.92 (9H, s); 1.30 (3H, d, J= 7 Hz); 1.40-1.60 (9H, complex); 1.78 (1H, m); 2.15 (3H, s); 2.40-2.60 (2H, m); 2.98 (1H, m); 4.38 (1H, d, J= 9 Hz); 4.46 (1H, d, J = 9 Hz); 4.92 (1H, pentet, J = 7 Hz); 6.98 (1H, d, J = 7.5 Hz); 7.02 (2H, m); 7.12 (3H, m); 7.20 (2H, m); 7.30 (1H, no s); 7.40 (1H, d, J= 7.5 Hz); 7.50 (1H, t, J= 7.5 Hz); 7.80 (3H, m); 8.00 (1H, no. s); 8.40 (1H, d, J= 7 Hz).

LRMS (thermospray) m/z = 664 (MNa+).

Found: C, 70.26; H, 7.59; N, 6.36.

C18H47N3O6 • 0.25 EtOAc required: C, 70.56; H, 7.44; N, 6.33%.

Preparation 27

1-(3-Carbamoylphenyl)-4-iodo-2-methylbenzene

[image]

Potassium hydroxide (1.21 g, 21.6 mmol) was added to a solution of 1-(3-cyanophenyl)-4-iodo-2-methylbenzene (1.27 g, 4.0 mmol) in ethanol (50 mL) and heated at reflux for 16 hours. After cooling, the mixture was diluted with water (100 ml) and extracted with ethyl acetate (3 x 50 ml). The combined organic extracts were dried (Na2SO4) and concentrated under reduced pressure to give the crude product (1.08 g). The crude product (540 mg) was purified by flash chromatography (gradient elution with dichloromethane:saturated methanolic ammonia solution 100:2.5, dichloromethane:ethyl acetate 1:1, and dichloromethane:ethyl acetate:acetic acid 100:100:4), and then azeotroped with ethanol, ethyl acetate and diethyl ether to give 1-(3-carbamoylphenyl)-4-iodo-2-methylbenzene (310 mg, 46%) as a yellow-brown solid.

T.t. 138-141 °C.

Rf 0.40 (dichloromethane: ethyl acetate: acetic acid = 50:50:2.5)

δH (400 MHz, DMSO) 2.18 (3H, s); 7.00 (1H, d, J= 8 Hz); 7.38 (1H, no s); 7.48 (2H, m); 7.62 (1H, d, J= 8 Hz); 7.72 (1H, s); 7.80 (1H, s); 7.86 (1H, d, J = 8 Hz); 8.00 (1H, no. s).

LRMS (thermospray) m/z = 355 (MNH4+).

Preparation 28

tert-butyl (3R)-3-[({({[(1S)-2-methoxy-1-phenylethyl]amino}carbonyl)-[(1S)-2-methyl]-1-propyl}amino)carbonyl] -6-[3-methyl-(phenyl)phenyl]hexanoate

[image]

a) According to the procedure for preparing preparation 19a, N-tert-butoxycarbonyl-L-valine (868 mg, 4.0 mmol) was reacted with (1S)-2-methoxy-1-phenylethylamine (604 mg, 4.0 mmol) for 72 hours. The mixture was concentrated under reduced pressure and partitioned between ethyl acetate (200 ml) and water (200 ml). The organic layer was washed with 5% aqueous citric acid solution (100 ml), saturated aqueous sodium bicarbonate solution (100 ml), dried (MgSO4) and concentrated under reduced pressure. The product was triturated with diisopropyl ether, filtered and dried to give (2S)-tert-(butoxycarbonyl)amino-N-[(1S)-2-methoxy-1-phenylethyl]-3-methylbutanamide (1.11 g, 79 %) in the form of a colorless solid.

T.t. 112-113 °C.

Rf 0.71 (dichloromethane:methanol:concentrated aqueous ammonia solution = 90:10:1).

δH (400 MHz, CDCl 3 ) 0.96 (6H, d, J= 7 Hz); 1.45 (9H, s); 2.18 (1H, m); 3.35 (3H, s); 3.64 (2H, m); 3.92 (1H, m); 5.00 (1H, no. s); 5.18 (1H, m); 6.60 (1H, m); 7.32 (5H, m).

LRMS (thermospray) m/z = 351 (MH+).

b) (2S)-tert-(butoxycarbonyl)amino-N-[(1S)-2-methoxy-1-phenylethyl]-3-methylbutanamide (1.11 g, 3.17 mmol) was dissolved in a mixture of anhydrous dichloromethane ( 15 ml) and dioxane (15 ml) and cooled to 4 °C. Hydrogen chloride was bubbled through the solution with stirring until a saturated solution was formed. After stirring for 4 hours at 4 °C, the solution was concentrated under reduced pressure. The residue was azeotroped with dichloromethane to give (2S)-amino-N-[(1S)-2-methoxy-1-phenylethyl]-3-methylbutanamide hydrochloride (948 mg, 104%) as a pale yellow foam.

δH (400 MHz, CDCl 3 ) 0.95 (3H, d, J = 7.5 Hz); 1.00 (3H, d, J= 7.5 Hz); 2.20 (1H, m); 3.15 (3H, s); 3.56 (1H, m); 3.66 (1H, m); 4.02 (1H, m); 5.10 (1H, m); 7.20-7.35 (3H, m); 7.40 (2H, d, J= 8 Hz); 7.90 (3H, no s); 8.18 (1H, d, J = 7.5 Hz).

LRMS (thermospray) m/z = 251 (MH+).

c) According to the procedure for preparing preparation 1, (2R)-2-[2-(tert-butoxy)-2-oxoethyl]pent-4-enoic acid (646 mg, 3.02 mmol) was reacted with (2S )-amino-N-[(1S)-2-methoxy-1-phenylethyl]-3-methylbutanamide hydrochloride (from b, above) (908 mg, 3.17 mmol) for 1 h at 4 °C, then 18 hours at 20 °C. The mixture was concentrated under reduced pressure and partitioned between ethyl acetate (100 ml) and saturated aqueous sodium bicarbonate (100 ml). The layers were separated and the aqueous layer was re-extracted with ethyl acetate (100 ml). The combined organic layers were dried (MgSO4) and concentrated under reduced pressure. The residue was purified by flash chromatography (eluting with dichloromethane:methanol 98:2) and tert-butyl (3R)-3-({[({[(1S)-2-methoxy-1-phenylethyl]amino}carbonyl)- (1S)-2-Methyl-1-propyl]amino}carbonyl)hex-5-enoate (1.08 g, 80%), as a white solid.

T.t. 155-157 °C.

Rf 0.36 (dichloromethane:methanol = 95:5).

δH (400 MHz, CDCl 3 ) 0.94 (3H, d, J= 7 Hz); 0.97 (3H, d, J= 7 Hz); 1.40 (9H, s); 2.15 (2H, m); 2.35 (2H, m); 2.65 (2H, m); 3.15 (3H, s); 3.65 (2H, d, J= 5Hz); 4.26 (1H, t, J=7.5); 4.96 (1H, d, J= 9Hz); 5.00 (1H, d, J= 1 BHz); 5.15 (1H, dt, J= 5 and 7 Hz); 5.66 (1H, m); 6.26 (1H, d, J = 7.5 Hz); 6.55 (1H, d, J= 7 Hz); 7.30 (5H, m).

LRMS (thermospray) m/z = 447 (MH+).

d) According to the procedure for preparing preparation 15a, tert-butyl (3R)-3-({[({[(1S)-2-methoxy-1-phenylethyl]amino}carbonyl)-(1S)-2-methyl-1 -propyl]amino}carbonyl)hex-5enoate (from c, above) (1.08 g, 2.42 mmol) was reacted with 4-iodo-2-methyl-1-phenylbenzene (preparation 29) (1, 07 g, 3.63 mmol) under palladium catalysis at reflux in acetonitrile for 18 hours. After cooling, the mixture was diluted with ethyl acetate (200 ml) and washed successively with 5% aqueous citric acid (50 ml), saturated aqueous sodium bicarbonate (50 ml), saturated brine (50 ml), dried (Na2SO4 ) and condensed under reduced pressure. The residue was purified by flash chromatography (gradient elution with dichloromethane:ethyl acetate = 30:1 to 3:1) and obtained tert-butyl (3R,5E)-3-({[({[(1S)-2-methoxy- 1-Phenylethyl]amino}carbonyl)-(1S)-2-methyl-1-propyl]amino}carbonyl)-6-[3-methyl-(4-phenyl)]hex-5-enoate, as a light brown foam ( 980 mg, 66 %).

Rf 0.35 (dichloromethane:ethyl acetate = 4:1).

δH (400 MHz, CDCl 3 ) 0.96 (3H, d, J = 7 Hz); 0.98 (3H, d, J= 7 Hz); 1.40 (9H, s); 2.18 (1H, m); 2.22 (3H, s); 2.36 (1H, m); 2.40-2.58 (2H, m); 2.66 (1H, m); 2.75 (1H, m); 3.35 (3H, s); 3.62 (2H, d, J = 4.5 Hz); 4.30 (1H, t, J = 7.5 Hz); 5.12 (1H, dt, J= 4.5 and 7 Hz); 6.10 (1H, m); 6.36 (1H, d, J = 7 Hz); 6.40 (1H, d, J=16 Hz); 6.55 (1H, d, J = 7.5 Hz); 7.10 (2H, m); 7.20-7.38 (9H, complex); 7.40 (2H, m).

Found: C, 74.43; H, 7.85; N 4.57.

C38H48N2O5 required: C, 74.48; H, 7.90; N 4.57%.

e) Tert-butyl (3R,5E)-3-({[({[(1S)-2-methoxy-1-phenylethyl]amino}carbonyl)-(1S)-2-methyl-1-propyl]amino solution }carbonyl)-6-[3-methyl-(4-phenyl)phenyl]hex-5-enoate (from d, above) (930 mg, 1.51 mmol) in ethanol (50 ml) was hydrogenated over 10%- of palladium on carbon (75 mg) at 3 bar at 20 °C for 17 hours. The residue was purified by flash chromatography (gradient elution with penta:ethyl acetate 10:1 to 2:1) and triturated with diethyl ether to give tert-butyl (3R)-3-({[({[(1S)-2-methoxy -1-phenylethyl]amino}carbonyl)-(1S)-2-methyl-1-propyl]amino}carbonyl)-6-[3-methyl-(4-phenyl)phenyl]hexanoate (748 mg, 81 %) in kind of colorless foam, which was used in example 21.

Rf 0.35 (pentane:ethyl acetate = 2:1).

δH (400 MHz, CDCl 3 ) 0.95 (3H, d, J= 7 Hz); 0.98 (3H, d, J= 7 Hz); 1.40 (9H, s); 1.42 (1H, m); 1.60 (2H, m); 1.70 (1H, m); 2.18 (1H, m); 2.22 (3H, s); 2.38 (1H, m); 2.45-2.66 (4H, complex); 3.32 (3H, s); 3.60 (2H, d, J= 4.5 Hz); 4.28 (1H, t, J = 7.5 Hz); 5.15 (1H, dt, J= 4.5 and 7 Hz); 6.30 (1H, d, J= 7.5 Hz); 6.55 (1H, d, J= 7 Hz); 6.98 (1H, d, J= 8 Hz); 7.00 (1H, s); 7.10 (1H, d, J= 8 Hz); 7.20-7.36 (8H, complex); 7.40 (2H, m).

Found: C, 74.39; H 8.24; N 4.59.

C38H50N2O5 required: C, 74.32; H 8.27; N 4.61 %.

Preparation 29

4-iodo-2-methyl-1-phenylbenzene

[image]

a) According to the procedure for preparing Preparation 25a, 2-bromo-5-nitrotoluene (15 g, 69.4 mmol) was treated with phenyl boric acid (12.7 g, 104.1 mmol) with palladium catalysis, using a 2M aqueous solution sodium carbonate as a base, at reflux in dimethoxyethane for 16 hours. After cooling, the mixture was separated, and the organic layer was diluted with diethyl ether and washed with water (2×). The organic layer was dried (Na2SO4) and concentrated under reduced pressure. The residue was purified by flash chromatography (gradient elution with hexane to hexane:ethyl acetate 95:5) to give 2-methyl-4-nitro-1-phenylbenzene as a yellow solid (12.7 g, 86 %).

Rf 0.33 (hexane:ethyl acetate = 95:5).

δH (300 MHz, CDCl 3 ) 2.40 (3H, s); 7.20-7.50 (6H, complex); 8.10 (1H, d, J= 8 Hz); 8.15 (1H, s).

b) 2-methyl-4-nitro-1-phenylbenzene (12.7 g, 60 mmol) was dissolved in ethanol (260 ml) and hydrogenated over 10% palladium on carbon (1.27 g) at 3 bar at 20 °C for 2 hours. The mixture was filtered through Arbocel filter aid washing with ethyl acetate. The filtrate was concentrated under reduced pressure to give 4-amino-2-methyl-1-phenylbenzene (10.49 g, 95%) as a purple-brown oil.

δH (400 MHz, CDCl 3 ) 2.20 (3H, s); 3.60 (2H, no s); 6.60 (2H, m); 7.05 (1H, d, J= 8 Hz); 7.30 (3H, m); 7.40 (2H, m).

c) A mixed solution of 4-amino-2-methyl-1-phenylbenzene (10.49 g, 57.2 mmol) in diiodomethane (100 ml) was treated with iso-amyl nitrite (27 ml, 200.2 mmol) for 5 minutes . The mixture was heated at 75 °C for 45 minutes and then at 65 °C for 1 hour. After cooling, the mixture was concentrated under reduced pressure (96 °C / 30 mmHg) and the residue was purified by flash chromatography (eluting with hexane) to give 4-iodo-2-methyl-1-phenylbenzene (11.6 g, 66 %) in the form of oil.

Rf 0.33 (hexane).

δH (400 MHz, DMSO-d6) 2.20 (3H, s); 6.98 (1H, d, J= 8 Hz); 7.30 (2H, m); 7.35 (1H, m); 7.45 (2H, m); 7.60 (1H, d, J= 8 Hz); 7.70 (1H, s).

Preparation 30

tert-butyl (3R)-3-({[({[(1S)-2-methoxy-1-phenylethyl]amino}carbonyl)-(1S)-2-phenylethyl]amino}carbonyl)-6-[3- methyl-(4-phenyl)phenyl]hexanoate

[image]

a) According to the procedure for preparing preparation 19a, N-tert-butoxycarbonyl-1-phenylalanine (1.06 mg, 4.0 mmol) was reacted with (1S)-2-methoxy-1-phenylethylamine (604 mg, 4 .0 mmol) for 72 hours. The mixture was concentrated under reduced pressure and partitioned into ethyl acetate (200 ml) and water (200 ml). The organic layer was washed with 5% aqueous citric acid solution (100 ml), saturated aqueous sodium bicarbonate solution (100 ml), dried (MgSO4) and concentrated under reduced pressure. The product was triturated with diisopropyl ether, filtered and dried, after which (2S)-tert-(butoxycarbonyl)amino-N-[(1S)-2-methoxy-1-phenylethyl]-3-phenylpropanamide (1.28 g, 81 %) in the form of white solid matter.

T.t. 133-134 °C.

Rf 0.76 (dichloromethane:methanol:concentrated aqueous ammonia solution = 90:10:1).

δH (400 MHz, CDCl 3 ) 1.40 (9H, s); 3.02 (1H, dd, J= 7 and 13 Hz); 3.16 (1H, dd, J= 6 and 13 Hz); 3.22 (3H, s); 3.40 (1H, m); 3.56 (1H, dd, J= 4.5 and 10 Hz); 4.38 (1H, m); 5.05 (2H, m); 6.45 (1H, d, J= 8 Hz); 7.18-7.36 (10H, complex).

LRMS (thermospray) m/z = 399 (MH+).

b) (2S)-tert-(butoxycarbonyl)amino-N-[(1S)-2-methoxy-1-phenylethyl]-3-phenylpropanamide (1.28 g, 3.2 mmol) was dissolved in a mixture of anhydrous dichloromethane ( 15 ml) and dioxane (15 ml) and cooled to 4 °C. Hydrogen chloride was bubbled through the solution with stirring until a saturated solution was formed. After stirring for 4 hours at 4 °C, the solution was concentrated under reduced pressure. The residue was azeotroped with dichloromethane to give (2S)-amino-N-[(1S)-2-methoxy-1-phenylethyl]-3-phenylpropanamide hydrochloride (1.11 g, 103%) as a pale yellow foam.

δH (400 MHz, CDCl3) 3.08 (1H, dd, J=13 and 10 Hz); 3.20 (3H, s); 3.32 (2H, m); 3.50 (1H, dd, J= 7.5 and 6.5 Hz); 4.44 (1H, m); 5.00 (1H, m); 7.18 (1H, m); 7.20 (2H, m); 7.25 (7H, complex); 7.64 (1H, d, J= 8 Hz); 7.80 (3H, no s).

LRMS (thermospray) m/z = 299 (MH+).

c) According to the procedure for preparing preparation 2, (2R)-2-[2-(tert-butoxy)-2-oxoethyl]pent-4-enoic acid (658 mg, 3.07 mmol) was reacted with (2S )-amino-N-[(1S)-2-methoxy-1-phenylethyl]-3-phenylpropanamide hydrochloride (from b, above) (1.08 mg, 3.23 mmol) for 1 h at 4 °C, and then 18 hours at 20 °C. The mixture was concentrated under reduced pressure and partitioned into ethyl acetate (100 ml) and saturated aqueous sodium bicarbonate solution (100 ml). The layers were separated and the aqueous layer was re-extracted with ethyl acetate (100 ml). The combined organic layers were dried (MgSO4) and concentrated under reduced pressure. The residue was triturated with diisopropyl ether to give tert-butyl (3S)-3-({[({[(1S)-2-methoxy-1-phenylethyl]amino}carbonyl)-(1S)-2-phenylethyl]amino} carbonyl)hex-5-enoate (1.35 g, 85 %) as a white solid.

T.t. 122-125 °C.

Rf 0.47 (dichloromethane:methanol = 95:5).

δH (400 MHz, CDCl 3 ) 1.40 (9H, s); 2.12 (1H, m); 2.26-2.40 (2H, m); 2.50-2.60 (2H, m); 3.00 (1H, dd, 8.5 and 14 Hz); 3.19 (1H, dd, 5.5 and 14 Hz); 3.20 (3H, s); 3.36 (1H, dd, 4.5 and 9.5 Hz); 3.50 (1H, dd, 4.8 and 9.5); 4.65 (1H, q, J = 7.5 Hz); 4.94-5.04 (3H, m); 5.60 (1H, m); 6.37 (2H, m); 7.18-7.35 (10H, complex).

LRMS (thermospray) m/z = 495 (MH+).

d) According to the procedure for preparing preparation 15a, tert-butyl (3S)-3-({[({[(1S)-2-methoxy-1-phenylethyl]amino}carbonyl)-(1S)-2-phenylethyl) was obtained ]amino}carbonyl)hex-5-enoate (from c, above) (1.35 g, 2.73 mmol) which was reacted with 4-iodo-2-methyl-1-methylbenzene (preparation 29) (1 .21 g, 4.09 mmol) with palladium catalysis, at reflux and in acetonitrile for 18 hours. After cooling, the mixture was diluted with ethyl acetate (200 ml) and washed with 5% aqueous citric acid solution (50 ml), saturated aqueous sodium bicarbonate solution (50 ml), saturated with salt (50 ml), dried (Na2SO4) and concentrated under reduced pressure. The residue was purified by flash chromatography (gradient elution with dichloromethane:ethyl acetate = 50:1 to 5:1) and tert-butyl (3R,5E)-3-({[({[(1S)-2-methoxy- 1-phenylethyl]amino}carbonyl)-(1S)-2-phenylethyl]amino}carbonyl)-6-[3-methyl-(4-phenyl)phenyl]hex-5-enoate, in the form of a pale brown foam (1, 1 g, 61 %).

Rf 0.35 (dichloromethane.ethyl acetate = 5:1).

δH (400 MHz, CDCl 3 ) 1.40 (9H, s); 2.22 (3H, s); 2.34 (1H, m); 2.40-2.56 (2H, m); 2.56-2.75 (2H, m); 3.00 (1H, dd, J= 7.5 and 14 Hz); 3.19 (1H, dd, J= 6 and 14 Hz); 3.20 (3H, s); 3.35 (1H, dd, J= 4.5 and 10 Hz); 3.50 (1H, dd, J= 4.5 and 10 Hz); 4.65 (1H, q, J = 7.5 Hz); 5.00 (1H, dt, J= 4.5 and 7.5 Hz); 6.06 (1H, dt, J= 7 and 14 Hz); 6.34 (1H, d, J = 7.5 Hz); 6.38 (1H, d, J=14 Hz); 6.44 (1H, d, J = 7.5 Hz); 7.10-7.38 (16H, complex); 7.40 (2H, m).

Found: C, 76.10; H 7.27; N 4.20.

C42H48N2O5 required: C, 76.33; H 7.32; N 4.24%.

e) Oopina tert-butyl (3R,5E)-3-({[({[(1S)-2-methoxy-1-phenylethyl]amino}carbonyl)-(1S)-2-phenylethyl]amino}carbonyl)- 6-[3-Methyl-(4-phenyl)phenyl]hex-5-enoate (1.03 g, 1.56 mmol) in ethanol (100 ml) was hydrogenated over 10% palladium on carbon (125 mg) at 3 bar at 20 °C for 18 hours. The mixture was filtered through an Arbocel filter aid and concentrated under reduced pressure. The residue was purified by flash chromatography (gradient elution with pentane:ethyl acetate = 10:1 to 2:1) and triturated with diethyl ether, after which tert-butyl (3R)-3-({[({[(1S)- 2-Methoxy-1-phenylethyl]amino}carbonyl)-(1S)-2-phenylethyl]amino)carbonyl)-6-[3-methyl-(4-phenyl)phenyl] hexanoate (980 mg, 95%) as colorless foam, which was used in example 22.

Rf 0.35 (pentane:ethyl acetate = 2:1).

δH (400 MHz, CDCl 3 ) 1.40 (9H, s); 1.42 (1H, m); 1.48-1.70 (3H, m); 2.20 (3H, s); 2.38 (1H, m); 2.56 (4H, m); 3.00 (1H, dd, J= 8 and 13 Hz); 3.20 (3H, s, and 1H, dd, J= 5.5 and 13 Hz, overlapping); 3.36 (1H, dd, J= 5 and 9 Hz); 3.50 (1H, dd, J= 5 and 9 Hz); 4.65 (1H, q, J= 8 Hz); 5.00 (1H, dt, J= 5 and 8 Hz); 6.38 (2H, d, J = 8 Hz); 6.95 (1H, d, J= 8 Hz); 7.00 (1H, s); 7.10 (1H, d, J= 8 Hz); 7.14-7.35 (13H, complex); 7.40 (2H, m).

Found: C, 76.03; H, 7.70; N, 4.25.

C42H50N2O5 required: C, 76.10; H, 7.60; N, 4.23%.

Preparation 31

tert-butyl (3R)-3-({[(1S)-2,2-dimethyl-1-{[(1R)-1-(4-pyridyl)ethylamino]carbonyl}propyl]amino}carbonyl)-6- [(3-Methyl-4-phenyl)phenyl]hexanoate

[image]

According to the procedure from example 25a, tert-butyl (3R)-3-({[(1S)-1-(carboxy)-2,2-dimethylpropyl]amino}carbonyl)-6-[(3-methyl-4-phenyl )phenyl]hexanoate (preparation 19c) (497 mg, 1.0 mmol) was reacted with (R)-(+)-(4-pyridyl)ethylamine (122 mg, 1.0 mmol) at room temperature for 2 ,5 hours, and then with the same treatment, tert-butyl (3R)-3-({[(1S)-2,2-dimethyl-1-{[(1R)-1-(4-pyridyl)ethylamino] was obtained carbonyl}propyl]amino}carbonyl)-6-[(3-methyl-4-phenyl)phenyl]hexanoate (460 mg, 77%) as a white solid.

Rf 0.21 (hexane:ethyl acetate = 1:2).

δH (400 MHz, CDCl 3 ) 1.01 (9H, s); 1.40 (9H, s); 1.45 (3H, d, J = 7.5 Hz); 1.50-1.65 (4H, m); 2.22 (3H, s); 2.38 (1H, m); 2.50-2.65 (4H, m); 4.20 (1H, d, J= 9 Hz); 5.04 (1H, pentet, J = 7.5 Hz); 6.15 (1H, d, J = 7.5 Hz); 6.45 (1H, d, J= 9 Hz); 7.00 (2H, m); 7.14 (3H, m); 7.30 (3H, m); 7.38 (2H, m); 8.45 (2H, d, J = 7.5 Hz).

LRMS (thermospray) m/z = 600 (MH+).

Example 38 / Preparation 32

(3R)-3-({[(1S)-2,2-dimethyl-1-{[(1R)-1-(3-pyridyl)ethylamino]carbonyl}propyl]amino}carbonyl)-6-[(3 -methyl-4-phenyl)phenyl]hexanoic acid

[image]

a) According to the procedure from example 25a, tert-butyl (3R)-3-({[(1S)-1-(carboxy)-2,2-dimethylpropyl]amino}carbonyl)-6-[(3-methyl-4 -phenyl)phenyl]hexanoate (preparation 19c) (497 mg, 1.0 mmol) was reacted with (R)-(+)-1-(3-pyridyl)ethylamine (122 mg, 1.0 mmol) at at room temperature for 2.5 hours, and then by the same treatment and purification by flash chromatography (with gradient elution with hexane:ethyl acetate 4:1 to 100% ethyl acetate) tert-butyl (3R)-3-({[(1S)) was obtained -2,2-dimethyl-{[(1R)-1-(3-pyridyl)ethylamino]carbonyl}popyl]amino}carbonyl)-6-[3-methyl-(4-phenyl)phenyl]hexanoate (537 mg, 89 %) in the form of colorless foam.

Rf 0.33 (ethyl acetate).

δH (400 MHz, DMSO-d6) 0.90 (9H, s); 1.02 (3H, d, J = 7.5 Hz); 1.34 (9H, s); 1.20-1.34 (4H, m); 2.14 (3H, s); 2.20 (1H, m); 2.30-2.50 (3H, m); 2.80 (1H, m); 4.30 (1H, d, J= 9.5 Hz); 4.95 (1H, pentet, J= 7.5 Hz); 6.92 (1H, d, J= 8 Hz); 7.00 (2H, m); 7.18 (1H, m); 7.24 (2H, m); 7.32 (1H, m); 7.40 (2H, m); 7.60 (1H, d, J= 8 Hz); 7.75 (1H, d, J = 9.5 Hz); 8.35 (1H, m); 8.42 (1H, d, J = 7.5 Hz); 8.46 (1H, s).

LRMS (thermospray) m/z = 600 (MH+).

b) Hydrogen gas is bubbled through a solution of tert-butyl (3R)-3-({[(1S)-2,2-dimethyl-{[(1R)-1-(3-pyridyl)ethylamino]carbonyl}propyl]amino }carbonyl)-6-[3-methyl-(4-phenyl)phenyl]hexanoate (285 mg, 0.47 mmol) in dioxane (15 ml) under nitrogen at 0 °C, until satd. The solution was stirred for 3 hours at 0 °C and then concentrated under reduced pressure. The residue was dissolved in methanol (3 ml), then diethyl ether (3 ml) was added, and the mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was dissolved in methanol (3 ml), toluene (3 ml) was added, and the mixture was concentrated under reduced pressure. Finally, diethyl ether (5 ml) was added to the sonicated residue to give a white solid which was filtered and dried to give the title compound as a white solid (338 mg, 65 %).

Rf 0.06 (ethyl acetate:hexane:acetic acid = 50:50:1).

δH (400 MHz, CD3OD) 1.02 (9H, s); 1.40-1.60 (4H, m); 1.54 (3H, d, J = 7.5 Hz); 2.16 (3H, s); 2.30-2.58 (3H, m); 2.62 (1H, m); 2.90 (1H, m); 4.20 (1H, m); 5.15 (1H, pentet, J= 7.5 Hz); 6.90 (1H, d, J= 8 Hz); 6.98 (2H, m); 7.22 (2H, d, J= 8 Hz); 7.32 (1H, m); 7.40 (2H, d, J= 8 Hz); 7.80 (1H, m); 7.85 (1H, d, J = 9.5 Hz); 8.50 (2H, m); 8.80 (2H, m).

LRMS (thermospray) m/z = 544 (MH+).

Found: C, 65.78; H, 7.26; N 6.72.

C33H41N3O4 • HCl • 1.25 H2O requirements: C, 65.74; H, 7.63; N 6.76 %.

Preparation 33

(2R)-N-[(1S)-1-(carboxy)-2,2-dimethylpropyl]-2-[(4S)-2,2-dimethyl-5-oxo-1,3-dioxolan-4-yl ]-5-[(3-methyl-4-phenyl)phenyl]pentanamide

[image]

a) According to the procedure for making preparation 2, (4S)-4-[(1R)-1-carboxy-1-but-3-enyl]-2,2-dimethyl-1,3-dioxolan-5-one (preparation 14b) (2.39 g, 11.20 mmol) was reacted with L-tert-leucine benzyl ester hydrochloride (3.03 g, 11.76 mmol) for 1 hour at 4 °C and then for 17 hours at 20 °C. The mixture was concentrated under reduced pressure, dissolved in ethyl acetate (200 ml), washed with 0.5 M aqueous dihydrogen phosphate solution (2 x 200 ml), 5% saturated aqueous sodium bicarbonate solution (200 ml) and water, dried ( MgSO4) and condensed under reduced pressure. The residue was purified by flash chromatography (gradient elution with hexane:ethyl acetate 10:1 to 1:1) to give (2R)-N-[(1S)-1-[(benzyloxy)carbonyl]-2,2-dimethylpropyl] -2-[(4S)-2,2-dimethyl-5-oxo-1,3-dioxolan-4-yl]pent-4-enamide (4.06 g, 87%), as a white solid.

T.t. 67-70 °C.

Rf 0.30 (hexane:diethyl ether = 1:1).

δH (400 MHz, DMSO-d6) 0.95 (9H, s); 1.45 (3H, s); 1.54 (3H, s); 2.30 (1H, m); 2.45 (1H, m); 3.00 (1H, m); 4.02 (1H, d, J= 8 Hz); 4.50 (1H, d, J= 8 Hz); 4.85 (1H, d, J= 8 Hz); 5.00 (1H, d, J=16 Hz); 5.10 (2H, s); 5.60 (1H, m); 7.34 (5H, m); 8.12 (1H, d, J= 8 Hz).

LRMS (thermospray) m/z = 418 (MH+).

Found: C, 66.13; H, 7.50; N, 3.36.

C23H31NO6 required: C, 66.17; H, 7.48; N, 3.35%.

b) Palladium acetate (58 mg, 0.24 mmol) was added to a stirred solution of (2R)-2-[(4S)-2,2-dimethyl-5-oxo-1,3-dioxolan-4-yl]-N -{(1S)-2,2-dimethyl-1-[(benzyloxy)carbonyl]propyl}pent-4-enamide (2.0 g, 4.8 mmol), 4-iodo-2-methyl-1-phenylbenzene (preparation 29) (2.1 g, 7.20 mmol) and N-ethylmorpholine (920 µl, 7.20 mmol) in anhydrous acetonitrile (15 ml) under nitrogen. The mixture was flushed with nitrogen, and then heated under reflux. After 1 hour, due to precipitation of palladium, palladium acetate (58 mg, 0.24 mmol) was added and refluxing was continued. After another 2 hours, another 58 mg of palladium acetate was added. After another 2 hours, tri-(2-methylphenyl)phosphine (304 mg, 1.0 mmol) and palladium acetate (58 mg, 0.24 mmol) were added, and the mixture was refluxed for 2 hours, after which three -(2-methylphenyl)phosphine (304 mg, 1.0 mmol) and palladium acetate (58 mg, 0.24 mmol), and the mixture was refluxed for another 8 hours. The reaction mixture was cooled, diluted with ethyl acetate (200 ml) and water (200 ml), and filtered through an Arbocel filter aid to remove palladium. The layers were separated and the organic layer was washed with water (2 x 100 ml), dried (Na2SO4) and concentrated under reduced pressure. The crude product was purified by flash chromatography. First, a yellow resin was obtained by chromatography with gradient elution with dichloromethane:ethyl acetate = 50:1 to 15:1). Further purification by flash chromatography (with gradient elution with hexane:diethyl ether 10:1 to 1:1) and trituration with pentane:diethyl ether yielded (2R,4E)-N-{(1S)-1-[(benzyloxy)carbonyl]-2 ,2-dimethylpropyl}-2-[(4S)-2,2-dimethyl-5-oxo-1,3-dioxolan-4-yl]-5-[(3-methyl-4-phenyl)phenyl]pent- 4-enamide as a white solid (710 mg, 25 %).

T.t. 115-118 °C.

Rf 0.30 (hexane:ethyl acetate = 1:1).

δH (300 MHz, CDCl 3 ) 0.96 (9H, s); 1.54 (3H, s); 1.60 (3H, s); 2.24 (3H, s); 2.65-2.90 (3H, m); 4.50 (1H, d, J= 9.5 Hz); 4.60 (1H, d, J = 7.5 Hz); 5.0 (1H, d, J= 9.5 Hz); 5.3 (1H, d, J= 9.5 Hz); 6.20 (1H, m); 6.44 (1H, d, J = 9.5 Hz); 6.52 (1H, d, J=16 Hz); 7:10-7.42 (13H, complex).

LRMS (thermospray) m/z = 584 (MH+).

Found: C, 73.96; H, 7.02; N, 2.41.

C36H41NO6 requirements: C, 74.08; H, 7.08; N, 2.40%.

c) Solution of (2R,4E)-N-{(1S)-1-[(benzyloxy)carbonyl]-2,2-dimethylpropyl}-2-[(4S)-2,2-dimethyl-5-oxo-1 ,3-dioxolan-4-yl]-5-[(3-methyl-4-phenyl)phenyl]pent-4-enamide (1.24 g, 2.12 mmol) in ethanol (150 ml) was hydrogenated over 10 % palladium on carbon (100 mg) at 3 bar at room temperature for 6 hours. The mixture was filtered through an Arbocel filter aid and concentrated under reduced pressure. The residue was azeotroped with diethyl ether to give (2R)-N-[(1S)-1-(carboxy)-2,2-dimethylpropyl]-2-[(4S)-2,2-dimethyl-5-oxo-1 ,3-dioxolan-4-yl]-5-[(3-methyl-4-phenyl)phenyl]pentanamide (1.08 g, 103 %) as a colorless foam.

Rf 0.40 (hexane:ethyl acetate:acetic acid = 50:50:1).

δH (400 MHz, DMSO-d6) 0.95 (9H, s); 1.46 (3H, s); 1.55 (3H, s); 1.50-1.66 (3H, m); 1.80 (1H, m, ); 2.20 (3H, s); 2.50 (1H, m); 2.60 (1H, m); 3.00 (1H, m); 4.15 (1H, d, J= 8 Hz); 4.50 (1H, d, J= 8 Hz); 7.05 (3H, m); 7.30 (3H, m); 7.40 (2H, m); 7.95 (1H, d, J= 8 Hz).

Found: C, 69.50; H, 7.69; N, 2.69.

C29H37NO6 • 0.3 H2O requirements: C, 69.52; H, 7.56; N, 2.80%.

Preparation 34

(2R)-N-[(1S)-1-(carboxy)-2,2-dimethylpropyl]-2-[(4S)-2,2-dimethyl-5-oxo-1,3-dioxolan-4-yl ]-5-[(3'-methoxy-2-methylbiphen-4-yl)propyl]pentanamide

[image]

a) According to the procedure for making preparation 15a, (2R)-N-[(1S)-1-[(benzyloxy)carbonyl]-2,2-dimethylpropyl]-2-[(4S)-2,2-dimethyl-5 -oxo-1,3-dioxolan-4-yl]pent-4-enamide (preparation 32a) (1.0 g, 2.4 mmol) was reacted with 4-iodo-1-(3-methoxyphenyl)- with 2-methylbenzene (preparation 20) (1.16 g, 3.59 mmol) under palladium catalysis and under reflux in acetonitrile for 16 hours. After cooling, the mixture was partitioned between ethyl acetate (3 x 75 mL) and saturated aqueous sodium bicarbonate (75 mL). The combined organic solutions were dried (Na2SO4) and concentrated under reduced pressure. The residue was purified by flash chromatography (eluting with dichloromethane:diethyl ether = 97.5:2.5) and obtained (2R,4E)-N-{(1S)-1-[(benzyloxy)carbonyl]-2,2- dimethylpropyl}-2-[(4S)-2,2-dimethyl-5-oxo-1,3-dioxolan-4-yl]-5-[(3'-methoxy-2-methylbiphen-4-yl)pent- 4-enamide as a solid (1.08 mg, 74 %). 1H NMR suggested that two alkene isomers were also present. The alkene mixture was carried to the next step (see b, below).

Rf 0.44 (dichloromethane:diethyl ether = 95:5).

δH (400 MHz, CDCl3) (for (4E) isomer) 0.95 (9H, s); 1.52 (3H, s); 1.60 (3H, s); 2.25 (3H, s); 2.54-2.90 (3H, m); 3.80 (3H, s); 4.50 (1H, d, J= 9.5 Hz); 4.60 (1H, d, J= 5 Hz); 4.98 (1H, d, J=12 Hz); 5.07 (1H, d, J=12 Hz); 6.20 (1H, m); 6.45 (1H, d, J= 9 Hz); 6.50 (1H, d, J= 16 Hz); 6.86 (3H, m); 7.12-7.40 (9H, complex).

LRMS (thermospray) m/z = 613 (MH+); 630 (MNH4+).

b) Solution of (2R,4E)-N-{(1S)-1-[(benzyloxy)carbonyl]-2,2-dimethylpropyl}-2-[(4S)-2,2-dimethyl-5-oxo-1 ,3-dioxolan-4-yl]-5-[(3'-methoxy-2-methylbiphen-4-yl)phenyl]pent-4-enamide (534 mg, 0.87 mmol) in ethanol (75 ml) hydrogenated is over 10% palladium on carbon (50 mg) at 3 bar at 20 °C for 5.75 hours. The mixture was filtered through an Arbocel filter aid, concentrated under reduced pressure and azeotroped with ethyl acetate, after which (2R)-N-[(1S)-1-(carboxy)-2,2-dimethylpropyl]-2-[( 4S)-2,2-dimethyl-5-oxo-1,3-dioxolan-4-yl]-5-[(3'-methoxy-2-methylbiphen-4-yl)propyl]pentanamide (384 mg, 61 % ) in the form of a white solid, which was used in examples 30 and 31 without purification.

δH (400 MHz, DMSO-d6) 0.96 (9H, s); 1.45 (3H, s); 1.55 (3H, s); 1.50-1.65 (3H, m); 1.80 (1H, m); 2.20 (3H, s); 2.50 (1H, m); 2.60 (1H, m); 2.96 (1H, m); 3.78 (3H, s); 4.15 (1H, d, J= 9 Hz); 4.50 (1H, d, J= 9 Hz); 6.85 (3H, m); 7.00 (1H, m); 7.05 (2H, m); 7.32 (1H, t, J= 8 Hz); 7.96 (1H, d, J= 9 Hz); 12.50 (1H, no. s).

LRMS (thermospray) m/z = 526 (MH+).

Claims (38)

1. The substance represented by the compound of formula (I) [image] or a pharmaceutically acceptable salt thereof, or a solution or any whole, indicated that R 1 is H, OH, C 1-4 alkyl, C 1-4 alkoxy or C 1-4 alkenyl; R2 is C1-6 alkyl, optionally substituted with fluoro, indolyl, imidazolyl, SO2(C1-4 alkyl), C5-7 cycloalkyl, or optionally protected by the group OH, SH, CONH2, CO2H, NH2 or NHC(=NH)NH2, C5-7 cycloalkyl, optionally substituted with C1-6 alkyl, or benzyl, optionally substituted with optionally protected OH, C1-6 alkoxy, benzyloxy or benzylthio, where, if desired, the protective groups for the mentioned OH, SH, CONH2, NH2 and NHC(=NH)NH2 groups are selected from C1-6 alkyl, benzyl, C1-6 alkanoyl, and where optionally selected protecting groups for CO2H are selected from C1-6 alkyl or benzyl; R 3 , R 5 and R 6 are independently selected from H and F; R 4 is CH 3 , Cl or F; X is HO or HONH; Y is a direct bond or O; Z is one group of formula (a) [image] where R10 is C1-4 alkyl, C1-4 alkoxymethyl, hydroxy(C2-4 alkyl), carboxy(C1-4 alkyl) or (amino or dimethylamino) C2-4 alkyl, R 11 is phenyl, naphthyl or pyridyl, optionally substituted with up to three substituents, independently selected from halo and methyl; or (b) [image] R14 is H, OH, CH3 or halo, Ar is a group of formula (c), (d) or (e) [image] in which A is N or CR12, B is N or CR13, provided that A and B are both not N; R 7 and R 9 are each independently H or F; R 8 , R 12 and R 13 are each independently H, CN, C 1-6 alkyl, hydroxy(C 1-6 alkyl), hydroxy(C 1-6 )alkoxy, C 1-6 alkoxy(C 1-4 ) alkoxy, (amino or dimethylamino)C 1 -6 alkyl, CONH2 OH, halo, C1-6 alkoxy, (C1-6 alkoxy)methyl, piperazinylcarbonyl, piperidinyl, C(NH2)=NOH or C(=NH)NHOH, provided that from R8, R12 and R13 at least two H. 2. A substance according to claim 1, characterized in that R1 is H, OH, C1-4 alkyl or C1-4 alkoxy. 3. A substance according to any of the preceding claims, characterized in that R2 is C1-4 alkyl, optionally substituted with indolyl, C1-6 alkylthio, SO2(C1-4 alkyl), C5-7 cycloalkyl, OH or SH, C5- 7 cycloalkyl optionally substituted with C1-6 alkyl, or R2 is benzyl. 4. A substance according to any of the preceding claims, characterized in that Z is a group of formula (a) [image] wherein R 10 is C 1-4 alkyl, C 1-4 alkoxymethyl or hydroxy(C 2-4 alkyl), and R 11 is phenyl or pyridyl, optionally substituted with up to three substituents, independently selected from halo and methyl; or Z is [image] . 5. A substance according to any of the preceding claims, characterized in that R3 is H. 6. A substance according to any one of the preceding claims, characterized in that R 4 is F when Y is O, and R 4 is Cl or CH 3 when Y is a direct bond. 7. A substance according to any of the preceding claims, characterized in that R5 is H. 8. A substance according to any of the preceding claims, characterized in that R6 is H. 9. A substance according to any of the preceding claims, characterized in that Ar is a group of formula (c) [image] where A is CR12 and B is CR13; R 7 and R 9 are each independently H or F; R 8 and R 13 are each independently H, F, Cl, CN, CONH 2 , CH 3 or OCH 3 ; and R12 is H, C1-6 alkyl, CN, hydroxy(C2-6 alkyl), (amino or dimethylamino)C2-6 alkyl, CONH2, OH, halo, C1-6 alkoxy, (C1-6 alkoxy)methyl, piperazinylcarbonyl, piperidinyl, C(NH2)NOH or C(=NH)NHOH. 10. A substance according to any of the preceding claims, characterized in that R1 is H, OH, n-propyl or ethoxy. 11. A substance according to any of the preceding claims, characterized in that R2 is C1-6 alkyl, optionally substituted with OH, SO2(C1-4 alkyl) or C5-7 cycloalkyl, cyclohexyl, optionally substituted with C1-4 alkyl; or R 2 is benzyl. 12. A substance according to any of the preceding claims, characterized in that group Z is of the formula (a) [image] wherein R 10 is C 1-4 alkyl, C 1-4 alkoxymethyl or hydroxy(C 2-4 alkyl), and R 11 is phenyl, pyridin-4-yl or pyridin-3-yl; or Z is [image] . 13. A substance according to any of the preceding claims, characterized in that Ar is a group of formula (c) [image] wherein A is CR12, B is CR13, and R7, R8, and R9 are H. 14. A substance according to any of the preceding claims, characterized in that R1 is H. 15. A substance according to any of the preceding claims, characterized in that R2 is cyclohexylmethyl, isopropyl, 1-methylcyclohexyl, t-butyl, C(CH3)2SO2CH3, benzyl or C(CH3)2OH. 16. A substance according to any of the preceding claims, characterized in that Z is a group of formula (a) [image] wherein R 10 is CH 3 , CH 2 OCH 3 , or CH 2 OH, and R 11 is phenyl, pyridin-4-yl or pyridin-3-yl; or is Z [image] . 17. A substance according to any of the preceding claims, characterized in that Ar is a group of formula (c) [image] where A is CR12, B is CR13, and R7, R8 and R9 are H; R 12 is H, C 1-4 alkyl, CN, hydroxy(C 2-4 alkyl), (amino or dimethylamino)C 2-6 alkyl, CONH 2 , OH, halo, C 1-6 alkoxy, (C 1-6 alkoxy)methyl, C( NH2)=NOH or C(=NH)NHOH; and R 13 is H, OCH 3 , CN, CONH 2 , CH 3 , or F. 18. A substance according to any of the preceding claims, characterized in that R2 is isopropyl, t-butyl or benzyl. 19. A substance according to any of the preceding claims, characterized in that Z is a group of the formula [image] . 20. A substance according to any of the preceding claims, characterized in that Ar is phenyl, 3-methoxyphenyl, 4-cyanophenyl, 3-cyanophenyl, 3-carbamoylphenyl or 4-hydroxyamidinophenyl. 21. A substance according to any of the preceding claims, characterized in that R2 is t-butyl. 22. A substance according to any of the preceding claims, characterized in that Ar is phenyl or 3-methoxyphenyl. 23. Substance according to claim 1, characterized in that R 1 is H, OH, n-propyl or ethoxy; R2 is t-butyl, isopropyl or benzyl; Z is a formula group [image] wherein R 10 is CH 3 , CH 2 OCH 3 or CH 2 OH, and R 11 is phenyl, pyridin-4-yl or pyridin-3-yl; or Z is [image] ; R 3 is H; R 4 is CH 3 , Cl or F; R 5 is H; R 6 is H; and Ar is phenyl, 3-methoxyphenyl, 4-cyanophenyl, 3-cyanophenyl, 3-carbamoylphenyl or 4-hydroxyamidinophenyl. 24. A substance according to claim 1, characterized in that it is mainly defined by the references and Examples herein, including salts and solvates thereof. 25. The substance according to claim 1, characterized in that it is selected from the compounds listed in Examples 3, 4, 8, 14, 15, 16, 22, 29, 30, 31 and 32, or their salts. 26. Pharmaceutical preparation, characterized in that it contains a substance according to any of the preceding claims, mixed with a compatible additive, diluent or carrier. 27. A substance according to any one of the preceding claims 1 to 25, characterized in that it is used as a medicine. 28. A substance according to any one of claims 1 to 25, characterized in that it is used in the production of drugs for the treatment of conditions caused by MMP, especially MMP-3 and/or MMP-12 and/or MMP-13. 29. A treatment method for preventing conditions caused by MMP, especially MMP-3 and/or MMP-12 and/or MMP-13, characterized in that it comprises administering an effective amount of a substance according to one of claims 1 to 25, or its preparations according to a claim 26. 30. The process for the production of the substance according to claim 1, where X is OH, characterized in that it comprises the transformation of the compound of formula (II) [image] where X1 is a group capable of being transformed into carboxy, and the other ingredients are as defined in claim 1, such as the hydrolysis of compound (II) in which X1 is half of CO2 (t-butyl or methyl). 31. The process for producing a substance according to claim 1, in which X is NHOH and R1, R2, R3, R4, R5, R6, Y, Ar and Z are as defined in claim 1, characterized by the fact that it comprises the reaction of the compound of formula (I ) according to claim 1 in which X is OH, with hydroxylamine or a derivative of hydroxylamine. 32. The process for making a substance according to claim 1, in which X is NHOH, and R1, R2, R3, R4, R5, R6, Y, Ar and Z are as defined in claim 1, characterized by including the reaction of compounds of the formula (I) in which X is OH, with O-hydroxylamine or its salt, in the presence of a peptide coupling agent, to obtain a compound of formula (III) [image] which is followed by treatment with ammonium formate in the presence of a suitable catalyst. 33. A process for the production of a substance according to claim 1, wherein X is NHOH, R1 is OH, and R2, R3, R4, R5, R6, Y, Ar, and Z are as defined in Claim 1, characterized by comprising the reaction compound of formula (IV) with hydroxylamine or its salt [image] . 34. A process for the production of a substance according to claim 1 in which X is OH and R1, R2, R3, R4, R5, R6, Y, Ar and Z are as defined in claim 1, characterized in that it comprises cross-linking a compound of the formula (V ) with the compound of formula (VI) [image] where X2 is a protected acid, such as CO2CH3 or CO2 (t-butyl) group, and LG is a cross-linked leaving group such as I, Br or OSO2CF3, and R1, R2, R3, R4, R5, R6, Y, Ar and Z are as defined in claim 1, in the presence of a catalyst, for the production of one or more compounds of formulas (VIIa) and/or (VIIb) in which X2 is defined as above for compounds of formula (V), and R1, R2, R3, R4, R5, R6, Y, Ar and Z are as defined in claim 1, [image] which is followed by the reduction of the oleophilic half and the deprotection of the protected half of the acid X2. 35. Compound of formula (II) [image] characterized in that X1, R1, R2, R3, R4, R5, R6, Y, Ar and Z are as defined in claim 30, or salts thereof, or solvates of any entity. 36. Compound of formula (III) [image] characterized in that R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , Y, Ar and Z are as defined in claim 32, or salts thereof, or solvates of any entity. 37. Compound of formula (IV) [image] characterized in that R 2 , R 3 , R 4 , R 5 , R 6 , Y, Ar and Z are as defined in claim 33, or salts thereof, or solvates of any entity. 38. Compound of formula (VIIa) or (VIIb) [image] characterized in that X 2 , R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , Y, Ar and Z are as defined in claim 34, or salts thereof, or solvates of any moiety.