HK1227405A1 - Modulators of cellular adhesion - Google Patents

Abstract

The invention relates to modulators of cellular adhesion. Particularly, the present invention provides compounds having formula (I): and pharmaceutically acceptable derivatives thereof, wherein R1-R4, n, p, A, B, D, E, L and AR1 are as described generally and in classes and subclasses herein, and additionally provides pharmaceutical compositions thereof, and methods for the use thereof for the treatment of disorders mediated by the CD11/CD18 family of cellular adhesion molecules (e.g., LFA-1).

Description

Cell adhesion modulators

The application is a divisional application of international application PCT/US2004/036942 with application number 200480039802.8 and invented name of "cell adhesion regulator" entering China at 7/4 of 2006.

Priority

Priority is claimed for this application from U.S. provisional application 60/560,517 filed on 8.4.2004 and 60/517,535 filed on 5.11.2003, which applications are incorporated herein by reference in their entirety.

Technical field and background

Studies conducted over the past decade have helped elucidate the molecular events involved in intercellular interactions in the body, particularly those involving the movement and activation of cells in the immune system. See generally Springer, T.Nature,1990,346, 425-. Cell surface proteins, particularly cell adhesion molecules ("CAMs") and "leukocyte integrins" (leukointetgrins), including LFA-1, MAC-1 and gp150.95 (referred to as CD18/CD11a, CD18/CD11b, and CD18/CD11c, respectively), have accordingly been the subject of drug development with the aim of interfering with the process of leukocyte extravasation to the site of injury and leukocyte migration to different targets. For example, it is presently believed that activation of integrins expressed structurally on leukocytes is preceded by tight ligand/receptor interactions between the integrin (e.g., LFA-1) and one or more different intercellular adhesion molecules (ICAMs) designated ICAM-1, ICAM-2, ICAM-3, or ICAM-4 expressed on the vascular endothelial cell surface and on other leukocytes, prior to leukocyte extravasation as a mandatory component of the inflammatory response. The interaction of CAM with leukocyte integrins is a critical step in the normal functioning of the immune system. Immune processes such as antigen presentation, T cell mediated cytotoxicity and leukocyte extravasation are all thought to require cell adhesion mediated by ICAMs interacting with leukocyte integrins. See generally Kishimoto, t.k.; rothlein; R.R.Adv.Pharmacol.1994,25, 117-.

Clearly, due to the role of the interaction of CAM and leukocyte integrins in the immune response, it is desirable to modulate these specific interactions to achieve the desired therapeutic effect (e.g., inhibit the interaction in the case of an overactive immune response). Importantly, it has been demonstrated that antagonism of the interaction between CAM and leukocyte integrin can be achieved by drugs directed against either component. In particular, blockade of a CAM such as, for example, ICAM-1, or a leukocyte integrin such as, for example, LFA-1, against either or both of these molecules effectively inhibits the inflammatory response. In vitro models of inflammation and immune responses inhibited by antibodies to CAM or leukocyte integrins include antigen or mitogen-induced lymphocyte proliferation, homotypic aggregation of lymphocytes, T cell-mediated cytolysis, and antigen-specific induced tolerance. The relevance of the in vitro assay is supported by in vivo assays using antibodies against ICAM-1 or LFA-1. For example, antibodies to LFA-1 can prevent thyroid transplant rejection and prolong the survival of cardiac xenografts in mice (Gorski, A., Immunology Today, 1994,15, 251-255). More significantly, antibodies against ICAM-1 have shown in vivo efficacy as anti-inflammatory agents in human diseases such as renal allograft rejection and rheumatoid Arthritis (Rothlein, R.R., Scharschmidt, L., Adhesion Molecules; Wegner, C.D., Ed., 1994,1-38, Cosami, C.B. et al, J.Immunol., 1990,144, 4604-.

As described above, the use of anti-LFA-1 or anti-ICAM-1 antibodies to antagonize this interaction has been investigated. In addition, LFA-1 or ICAM-1 peptides, fragments or peptide antagonists have been investigated (see, e.g., U.S. Pat. Nos. 5,149,780, 5,288,854, 5,340,800, 5,424,399, 5,470,953, PCT publication WO 90/03400, WO90/13316, WO90/10652, WO91/19511, WO92/03473, WO94/11400, WO95/28170, JP4193895, EP314863, EP362526, EP362531), and small molecule antagonists. For example, several small molecules that affect the interaction of CAM and leukocyte integrins have been described in the literature. Natural products isolated from Trichilia rubra roots were found to be inhibitory in vitro cell binding assays (Musza, L.L. et al, Tetrahedron,1994,50, 11369-11378). A range of molecules (boscheli, d.h. et al, j.med.chem., 1994,37,717 and boscheli, d.h. et al, j.med.chem., 1995,38, 4597-. Another series of molecules was also found to have oral activity in delayed type hypersensitivity reactions in rats (Sanfilippo, P.J. et al, J.Med.chem., 1995,38, 1057-1059). All of these molecules appear to act non-specifically, either by inhibiting transcription of ICAM-1 and other proteins, or act intramolecularly to inhibit leukocyte integrin activation by unknown mechanisms, and none appear to directly antagonize CAM interaction with leukocyte integrins.

Clearly, although several classes of compounds have been investigated for therapeutic applications, there is still a need to develop new therapies capable of modulating the interaction between CAM and leukocyte integrins. In particular, it would be desirable to develop therapies capable of selectively targeting (preferably inhibiting) the interaction between LFA-1 and ICAM-1 that are useful as therapeutics for immune and/or inflammatory diseases.

Disclosure of Invention

As mentioned above, there is still a need to develop new therapies that can modulate the interaction between CAM and leukocyte integrins. The present invention provides novel compounds of the general formula (I),

and pharmaceutical compositions thereof, as generally described and subdivided herein, which compounds are useful as modulators of the CD11/CD18 family of cell adhesion molecules. Thus, these compounds are useful, for example, in the treatment of various LFA-1 related diseases, including immune and/or inflammatory diseases.

In yet another aspect, the present invention provides a method of treating any disease mediated by the CD11/CD18 family of cell adhesion molecules, the method comprising administering to a subject in need thereof a therapeutically effective amount of a compound of the present invention.

Definition of

As used herein, the term "aliphatic" includes both saturated and unsaturated straight chain (i.e., unbranched) or branched aliphatic hydrocarbons, which are optionally substituted with one or more functional groups. As understood by those skilled in the art, "aliphatic" is intended herein to include, but is not limited to, alkyl, alkenyl, alkynyl moieties. Thus, as used herein, the term "alkyl" includes both straight and branched chain alkyl groups. Similar convention applies to other general terms such as "alkenyl", "alkynyl", and the like. In addition, as used herein, the terms "alkyl", "alkenyl", "alkynyl", and the like include both substituted and unsubstituted groups. In certain embodiments, "lower alkyl" as used herein is used to denote those alkyl groups (substituted, unsubstituted, branched or unbranched) having about 1-6 carbon atoms.

In certain embodiments, alkyl, alkenyl, and alkynyl groups useful in the present invention contain about 1-20 aliphatic carbon atoms. In certain other embodiments, the alkyl, alkenyl, and alkynyl groups useful in the present invention contain from about 1 to about 10 aliphatic carbon atoms. In other embodiments, alkyl, alkenyl, and alkynyl groups useful in the present invention contain from about 1 to about 8 aliphatic carbon atoms. In other embodiments, alkyl, alkenyl, and alkynyl groups useful in the present invention contain about 1-6 aliphatic carbon atoms. In other embodiments, alkyl, alkenyl, and alkynyl groups useful in the present invention contain about 1-4 aliphatic carbon atoms. Thus, exemplary aliphatic groups include, but are not limited to, for example, methyl, ethyl, n-propyl, isopropyl, allyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, sec-pentyl, isopentyl, tert-pentyl, n-hexyl, sec-hexyl moieties, and the like, which may additionally bear one or more substituents. Alkenyl groups include, but are not limited to, for example, ethenyl, propenyl, butenyl, 1-methyl-2-buten-1-yl, and the like. Representative alkynyl groups include, but are not limited to, ethynyl, 2-propynyl (propargyl), 1-propynyl, and the like.

As used herein, the term "cycloaliphatic" refers to a compound that combines the properties of aliphatic and cyclic compounds, including, but not limited to, monocyclic or polycyclic aliphatic hydrocarbons and bridged cycloalkyl compounds, which are optionally substituted with one or more functional groups. As understood by those skilled in the art, "alicyclic" is intended herein to include, but is not limited to, cycloalkyl, cycloalkenyl, and cycloalkynyl moieties, which are optionally substituted with one or more functional groups. Thus, exemplary alicyclic groups include, but are not limited to, for example, cyclopropyl, -CH₂-cyclopropyl, cyclobutyl, -CH₂-cyclobutyl, cyclopentyl, -CH₂-cyclopentyl, cyclohexyl, -CH₂Cyclohexyl, cyclohexenylethyl, cyclohexylethyl, norbornyl moieties and the like, which may additionally bear one or more substituents.

As used herein, the term "alkoxy" or "alkyloxy" refers to a saturated (i.e., O-alkyl) or unsaturated (i.e., O-alkenyl and O-alkynyl) group attached to the parent molecular moiety through an oxygen atom. In certain embodiments, the alkyl group contains from about 1 to about 20 aliphatic carbon atoms. In certain other embodiments, the alkyl group contains from about 1 to about 10 aliphatic carbon atoms. In other embodiments, the alkyl groups useful in the present invention contain from about 1 to about 8 aliphatic carbon atoms. In other embodiments, the alkyl group contains from about 1 to about 6 aliphatic carbon atoms. In other embodiments, the alkyl group contains about 1-4 aliphatic carbon atoms. Examples of alkoxy include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, tert-butoxy, neopentyloxy, n-hexyloxy, and the like.

As used herein, the term "thioalkyl" refers to a saturated (i.e., S-alkyl) or unsaturated (i.e., S-alkenyl and S-alkynyl) group attached to the parent molecular moiety through a sulfur atom. In certain embodiments, the alkyl group contains from about 1 to about 20 aliphatic carbon atoms. In certain other embodiments, the alkyl group contains from about 1 to about 10 aliphatic carbon atoms. In other embodiments, the alkyl groups useful in the present invention contain from about 1 to about 8 aliphatic carbon atoms. In other embodiments, the alkyl group contains from about 1 to about 6 aliphatic carbon atoms. In other embodiments, the alkyl group contains about 1-4 aliphatic carbon atoms. Examples of thioalkyl groups include, but are not limited to, methylthio, ethylthio, propylthio, isopropylthio, n-butylthio, and the like.

The term "alkylamino" refers to a group having the structure-NHR ', where R' is alkyl as defined herein. The term "aminoalkyl" refers to a compound having the structure NH₂The group of R '-, wherein R' is alkyl as defined herein. In certain embodiments, the alkyl group contains from about 1 to about 20 aliphatic carbon atoms. In certain other embodiments, the alkyl group contains from about 1 to about 10 aliphatic carbon atoms. In other embodiments, the alkyl groups useful in the present invention contain from about 1 to about 8 aliphatic carbon atoms. In other embodiments, the alkyl group contains from about 1 to about 6 aliphatic carbon atoms. In other embodiments, the alkyl group contains about 1-4 aliphatic carbon atoms. Examples of alkylamino include, but are not limited to, methylamino, ethylamino, isopropylamino, and the like.

The above-mentioned esters of the compounds of the present inventionSome examples of substituents for aliphatic (and other) moieties include, but are not limited to, aliphatic, alicyclic, heteroaliphatic, heterocyclic, aromatic, heteroaromatic, aryl, heteroaryl, alkylaryl, heteroalkylaryl, alkylheteroaryl, heteroalkylheteroaryl, alkoxy, aryloxy, heteroalkoxy, heteroaryloxy, alkylthio, arylthio, heteroalkylthio, heteroarylthio, F, C1, Br, I, -OH, -NO₂、-CN、-CF₃、-CH₂CF₃、-CHC1₂、-CH₂OH、-CH₂CH₂OH、-CH₂NH₂、-CH₂SO₂CH₃、-C(O)R_x、-CO₂(R_x)、-CON(R_x)₂、-OC(O)R_x、-OCO₂R_x、-OCON(R_x)₂、-N(R_x)₂、-S(O)₂R_x、-NR_x(CO)R_xWherein R is_xIndependently in each occurrence, includes, but is not limited to, aliphatic, alicyclic, heteroaliphatic, heterocyclic, aryl, heteroaryl, alkylaryl, alkylheteroaryl, heteroalkylaryl, or heteroalkylheteroaryl, wherein any of the aliphatic, alicyclic, heteroaliphatic, heterocyclic, alkylaryl, or alkylheteroaryl substituents described above and herein can be substituted or unsubstituted, branched or unbranched, saturated or unsaturated, and wherein any of the aryl or heteroaryl substituents described above and herein can be substituted or unsubstituted. Further examples of substituents which are generally applicable are illustrated by the specific embodiments shown in the examples described herein.

Generally, as used herein, the term "aromatic moiety" refers to a stable monocyclic or polycyclic unsaturated moiety, preferably having 3 to 14 carbon atoms, each of which may be substituted or unsubstituted. In certain embodiments, the term "aromatic moiety" refers to a planar ring having a p-orbital perpendicular to the plane of the ring at each ring atom and satisfying the huckel rule where the number of pi electrons in the ring is (4n +2) (where n is an integer). Monocyclic or polycyclic, unsaturated moieties that do not meet one or all of these aromaticity criteria are defined herein as "non-aromatic" and are included in the term "alicyclic".

Generally, as used herein, the term "heteroaromatic moiety" refers to a monocyclic or polycyclic unsaturated moiety preferably having 3 to 14 carbon atoms, each of which may be substituted or unsubstituted, and includes at least one heteroatom selected from O, S and N within the ring (i.e., in place of a ring carbon atom). In certain embodiments, the term "heteroaromatic moiety" refers to a planar ring that includes at least one heteroatom, has a p-orbital perpendicular to the plane of the ring at each ring atom, and satisfies the huckel rule where the number of pi electrons of the ring is (4n +2) (where n is an integer).

It is also to be understood that the aromatic moiety and the heteroaromatic moiety may be linked by an alkyl or heteroalkyl moiety, as defined herein, and thus also includes- (alkyl) aromatic moieties, - (heteroalkyl) heteroaromatic moieties. Thus, as used herein, the phrases "aromatic or heteroaromatic moiety" and "aromatic, heteroaromatic, - (alkyl) aromatic, - (heteroalkyl) heteroaromatic, and- (heteroalkyl) heteroaromatic" are interchangeable. Substituents include, but are not limited to, any of the aforementioned substituents that result in the formation of a stable compound, i.e., substituents described for the aliphatic moiety, or substituents for other moieties disclosed herein.

As used herein, the term "aryl" is not clearly distinguished from the ordinary meaning of the term in the art and refers to an unsaturated cyclic moiety that includes at least one aromatic ring. In certain embodiments, "aryl" refers to a monocyclic or bicyclic carbocyclic ring system having one or two aromatic rings, including, but not limited to, phenyl, naphthyl, tetrahydronaphthyl, 2, 3-indanyl, indenyl, and the like.

As used herein, the term"heteroaryl" is not clearly distinguished from the ordinary meaning of this term in the art and refers to a cyclic aromatic group having five to ten ring atoms, one of which is selected from S, O and N; zero, one or two ring atoms are an additional heteroatom independently selected from S, O and N, and the remaining ring atoms are carbon, which group is attached to the rest of the molecule via any ring atom, such as, for example, pyridyl, pyrazinyl, pyrimidinyl, pyrrolyl, pyrazolyl, imidazolyl, thiazolyl, trifluoromethyl, and the like,Azolyl radical, isoAn azole group, a thiadiazole group, a triazole group, a,Oxadiazolyl, thienyl, furyl, quinolinyl, isoquinolinyl, and the like.

It is to be understood that aryl and heteroaryl (including bicyclic aryl) groups may be unsubstituted or substituted, wherein substitution includes independently replacing one or more hydrogen atoms thereon with any one or more moieties including, but not limited to: aliphatic, alicyclic, heteroaliphatic, heterocyclic, aromatic, heteroaromatic, aryl, heteroaryl, alkylaryl, heteroalkylaryl, alkylheteroaryl, heteroalkylheteroaryl, alkoxy, aryloxy, heteroalkoxy, heteroaryloxy, alkylthio, arylthio, heteroalkylthio, heteroarylthio, F, C1, Br, I, -OH, -NO₂、-CN、-CF₃、-CH₂CF₃、-CHC1₂、-CH₂OH、-CH₂CH₂OH、-CH₂NH₂、-CH₂SO₂CH₃、-C(O)R_x、-CO₂(R_x)、-CON(R_x)₂、-CO(O)R_x、-OCO₂R_x、-OCON(R_x)₂、-N(R_x)₂、-S(O)R_x、-S(O)₂R_x、-NR_x(CO)R_xWherein R is_xIndependently in each occurrence include, but are not limited to, aliphatic, alicyclic, heteroaliphatic, heterocyclic, aromatic, heteroaromatic, aryl, heteroaryl, alkylaryl, alkylheteroaryl, heteroalkylaryl, or heteroalkylheteroaryl, wherein any of the aliphatic, alicyclic, heteroaliphatic, heterocyclic, alkylaryl, or alkylheteroaryl substituents described above and herein can be substituted or unsubstituted, branched or unbranched, saturated or unsaturated, and wherein any of the aromatic, heteroaromatic, aryl, heteroaryl, - (alkyl) aryl, or- (alkyl) heteroaryl substituents described above and herein can be substituted or unsubstituted. In addition, it is to be understood that any two adjacent groups together may represent a 4,5, 6, or 7 membered substituted or unsubstituted cycloaliphatic or heterocyclic moiety. Further examples of substituents which are generally applicable are illustrated by the specific embodiments shown in the examples described herein.

As used herein, the term "cycloalkyl" specifically refers to groups having from three to seven, preferably from three to ten carbon atoms. Suitable cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and the like, as in the case of aliphatic, alicyclic, heteroaliphatic, or heterocyclic moieties, which may be optionally substituted with substituents including, but not limited to: aliphatic, alicyclic, heteroaliphatic, heterocyclic, aromatic, heteroaromatic, aryl, heteroaryl, alkylaryl, heteroalkylaryl, alkylheteroaryl, heteroalkylheteroaryl, alkoxy, aryloxy, heteroalkoxy, heteroaryloxy, alkylthio, arylthio, heteroalkylthio, heteroarylthio, F, C1, Br, I, -OH, -NO₂、-CN、-CF₃、-CH₂CF₃、-CHC1₂、-CH₂OH、-CH₂CH₂OH、-CH₂NH₂、-CH₂SO₂CH₃、-C(O)R_x、-CO₂(R_x)、-CON(R_x)₂、-CO(O)R_x、-OCO₂R_x、-OCON(R_x)₂、-N(R_x)₂、-S(O)₂R_x、-NR_x(CO)R_xWherein R is_xIndependently in each occurrence, include, but are not limited to, aliphatic, alicyclic, heteroaliphatic, heterocyclic, aromatic, heteroaromatic, aryl, heteroaryl, alkylaryl, alkylheteroaryl, heteroalkylaryl, or heteroalkylheteroaryl, wherein any of the aliphatic, alicyclic, heteroaliphatic, heterocyclic, alkylaryl, or alkylheteroaryl substituents as described above and herein can be substituted or unsubstituted, branched or unbranched, saturated or unsaturated, and wherein any of the aryl or heteroaryl substituents as described above and herein can be substituted or unsubstituted. Further examples of substituents which are generally applicable are illustrated by the specific embodiments shown in the examples described herein.

As used herein, the term "heteroaliphatic" refers to an aliphatic moiety in which one or more carbon atoms in the main chain have been replaced with a heteroatom. Thus, heteroaliphatic refers to an aliphatic chain comprising one or more oxygen, sulfur, nitrogen, phosphorus, or silicon atoms in place of carbon atoms. The heteroaliphatic moiety may be linear or branched, saturated or unsaturated. In certain embodiments, a heteroaliphatic moiety is substituted by independently replacing one or more hydrogen atoms thereon with one or more moieties including, but not limited to, the following substituents: aliphatic, alicyclic, heteroaliphatic, heterocyclic, aromatic, heteroaromatic, aryl, heteroaryl, alkylaryl, alkylheteroaryl, alkoxy, aryloxy, heteroalkoxy, heteroaryloxy, alkylthio, arylthio, heteroalkylthio, heteroarylthio, F, C1, Br, I, -OH, -NO₂、-CN、-CF₃、-CH₂CF₃、-CHC1₂、-CH₂OH、-CH₂CH₂OH、-CH₂NH₂、-CH₂SO₂CH₃、-C(O)R_x、-CO₂(R_x)、-CON(R_x)₂、-CO(O)R_x、-OCO₂R_x、-OCON(R_x)₂、-N(R_x)₂、-S(O)₂R_x、-NR_x(CO)R_xWherein R is_xIndependently in each occurrence, include, but are not limited to, aliphatic, alicyclic, heteroaliphatic, heterocyclic, aromatic, heteroaromatic, aryl, heteroaryl, alkylaryl, alkylheteroaryl, heteroalkylaryl, or heteroalkylheteroaryl, wherein any of the aliphatic, alicyclic, heteroaliphatic, heterocyclic, alkylaryl, or alkylheteroaryl substituents as described above and herein can be substituted or unsubstituted, branched or unbranched, saturated or unsaturated, and wherein any of the aryl or heteroaryl substituents as described above and herein can be substituted or unsubstituted. Further examples of substituents which are generally applicable are illustrated by the specific embodiments shown in the examples described herein.

As used herein, the term "heterocycloalkyl", "heterocycle" or "heterocyclic" refers to a compound that combines the properties of heteroaliphatic and cyclic compounds, including, but not limited to, saturated and unsaturated monocyclic or polycyclic ring systems having 5 to 16 atoms, wherein at least one ring atom is a heteroatom selected from O, S and N (wherein the nitrogen and sulfur heteroatoms may optionally be oxidized), wherein the ring system is optionally substituted with one or more functional groups as defined herein. In certain embodiments, the term "heterocycloalkyl", "heterocycle" or "heterocyclic" refers to a non-aromatic 5-, 6-or 7-membered ring or polycyclic group in which at least one ring atom is a heteroatom selected from O, S and N (wherein the nitrogen and sulfur heteroatoms may be optionally oxidized), including but not limited to bi-or tricyclic groups, including fused six-membered rings having one to three heteroatoms independently selected from oxygen, sulfur and nitrogen, wherein (i) each 5-membered ring has 0 to 2 double bonds, each 6-membered ring has 0 to 2 double bonds, and each 7-membered ring has 0 to 3 double bonds, (ii) the nitrogen and sulfur heteroatoms may be optionally oxidized, (iii) the nitrogen heteroatom may be optionally quaternized(iii) ammonification, and (iv) the ring of any of the above heterocycles may be fused to an aryl or heteroaryl ring. Representative heterocycles include, but are not limited to, various heterocycles such as furyl, thiofuryl (thiofuryl), pyranyl, pyrrolyl, thienyl, pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl, substituted thienyl, pyridyl, etc,Azolyl group,Oxazolidinyl, isoAzolyl radical, isoOxazolidinyl, diAn azole group, a thiadiazole group, a triazole group, a,Oxadiazolyl, tetrazolyl, triazolyl, thiatriazolyl,Triazolyl, thiadiazolyl, thia,Oxadiazolyl, morpholinyl, thiazolyl, thiazolidinyl, isothiazolyl, isothiazolidinyl, dithiazolyl, tetrahydrofuranyl, and benzo-fused derivatives thereof. In certain embodiments, a "substituted heterocycle, or heterocycloalkyl or heterocycle" group is used, which as used herein refers to a heterocycle, or heterocycloalkyl or heterocyclic group as defined above, one, two or three hydrogen atoms on which are substituted with substituents including, but not limited to: aliphatic, alicyclic, heteroaliphatic, heterocyclic, aromatic, heteroaromaticAryl, heteroaryl, alkylaryl, heteroalkylaryl, alkylheteroaryl, heteroalkylheteroaryl, alkoxy, aryloxy, heteroalkoxy, heteroaryloxy, alkylthio, arylthio, heteroalkylthio, heteroarylthio, F, Cl, Br, I, -OH, -NO₂、-CN、-CF₃、-CH₂CF₃、-CHCl₂、-CH₂OH、-CH₂CH₂OH、-CH₂NH₂、-CH₂SO₂CH₃、-C(O)R_x、-CO₂(R_x)、-CON(R_x)₂、-CO(O)R_x、-OCO₂R_x、-OCON(R_x)₂、-N(R_x)₂、-S(O)₂R_x、-NR_x(CO)R_xWherein R is_xIndependently in each occurrence include, but are not limited to, aliphatic, alicyclic, heteroaliphatic, heterocyclic, aromatic, heteroaromatic, aryl, heteroaryl, alkylaryl, alkylheteroaryl, heteroalkylaryl or heteroalkylheteroaryl, wherein any of the aliphatic, alicyclic, heteroaliphatic, heterocyclic, alkylaryl, or alkylheteroaryl substituents as described above and herein can be substituted or unsubstituted, branched or unbranched, saturated or unsaturated, and wherein any of the aromatic, heteroaromatic, aryl or heteroaryl substituents as described above and herein can be substituted or unsubstituted. Further examples of substituents which are generally applicable are illustrated by the specific embodiments shown in the examples described herein.

In addition, it is to be understood that any of the cycloaliphatic or heterocyclic moieties described above and herein may include an aryl or heteroaryl moiety fused thereto. Further examples of substituents which are generally applicable are illustrated by the specific embodiments shown in the examples described herein.

As used herein, the terms "halo" and "halogen" refer to an atom selected from fluorine, chlorine, bromine, and iodine.

The term "haloalkyl" denotes an alkyl group as defined above having one, two, or three halogen atoms attached thereto, as exemplified by groups such as chloromethyl, bromoethyl, trifluoromethyl, and the like.

As used herein, the term "amino" refers to a primary amine (-NH)₂) Secondary amine (-NHR)_x) Tertiary amines (-NR)_xR_y) Or quaternary amines (-N)⁺R_xR_yR_z) Wherein R is_x、R_yAnd R_zIndependently an aliphatic, alicyclic, heteroaliphatic moiety, heterocyclic, aromatic moiety or heteroaromatic moiety as defined herein. Examples of amino groups include, but are not limited to, methylamino, dimethylamino, ethylamino, diethylamino, diethylaminocarbonyl, methylethylamino, isopropylamino, piperidino, trimethylamino, and propylamino.

As used herein, the term "acyl" refers to a group of the general formula-C (═ O) R, where R is an aliphatic, alicyclic, heteroaliphatic moiety, heterocyclic, aromatic moiety, or heteroaromatic moiety as defined herein.

As used herein, the term "sulfonamide" refers to the general formula-SO₂NR_xR_yWherein R is_xAnd R_yIndependently hydrogen, or an aliphatic, alicyclic, heteroaliphatic moiety, heterocyclic, aromatic moiety, heteroaromatic moiety, or acyl moiety as defined herein.

As used herein, the term "benzamido" refers to the general formula PhNR_xA group of (a) wherein R_xIs hydrogen or is an aliphatic, alicyclic, heteroaliphatic moiety, heterocyclic, aromatic moiety, heteroaromatic moiety, or acyl moiety as defined herein.

As used herein, the term "C_1-6By "alkylene" is meant a substituted or unsubstituted radical containing only carbon and hydrogen atoms, having from one to six carbon atoms, and having a free valence "-" at both ends of the radicalSubstituted, straight or branched chain saturated divalent radicals.

As used herein, the term "C_2-6Alkenylene "refers to a substituted or unsubstituted, straight or branched chain, unsaturated divalent radical containing only carbon and hydrogen atoms, having two to six carbon atoms, with a free valence" - "at both ends of the radical, where the unsaturation is present only as a double bond, and where a double bond may be present between the first carbon of the chain and the rest of the molecule.

As used herein, the terms "aliphatic," "heteroaliphatic," "alkyl," "alkenyl," "alkynyl," "heteroalkyl," "heteroalkenyl," "heteroalkynyl," and the like include substituted and unsubstituted, saturated and unsaturated, straight and branched chain radicals. Similarly, the terms "cycloaliphatic," "heterocyclic," "heterocycloalkyl," "heterocyclic," and the like include substituted and unsubstituted, saturated and unsaturated groups. In addition, the terms "cycloalkyl", "cycloalkenyl", "cycloalkynyl", "heterocycloalkyl", "heterocycloalkenyl", "heterocycloalkynyl", "aromatic", "heteroaromatic", "aryl", "heteroaryl", and the like, include both substituted and unsubstituted groups.

The term "protecting group" as used herein means that a specific functional group such as O, S, or N is temporarily blocked so that a reaction can be selectively performed at another active site of the polyfunctional compound. In a preferred embodiment, the protecting groups are reacted selectively in good yield to give a protected substrate that is stable to the intended reaction; the protecting group must be selectively removed in good yield by readily available, preferably non-toxic, reagents that do not attack other functional groups; the protecting group forms an easily cleavable derivative (more preferably does not generate a new stereocenter); the protecting group has minimal additional functionality to avoid additional reaction sites. As detailed herein, oxygen, sulfur, nitrogen, and carbon protecting groups may be used. For example, as detailed herein, in certain embodiments, certain exemplary oxygen protecting groups are used. Such oxygen protecting groups include, but are not limited to, methyl ether, substituted methyl ether (e.g., MOM (methoxymethyl ether), MTM (methylthiomethyl ether), BOM (benzyloxymethyl ether), PMBM, or MPM (p-methoxybenzyloxymethyl ether), to name a few); substituted ethyl ethers, substituted benzyl ethers, silyl ethers (e.g., TMS (trimethylsilyl ether), TES (triethylsilyl ether), TIPS (triisopropylsilyl ether), TBDMS (t-butyldimethylsilyl ether), tribenzylsilyl ether, TBDPS (t-butyldiphenylsilyl ether), to name a few); esters (e.g., formates, acetates, benzoates (Bz), trifluoroacetates, dichloroacetates, to name a few), carbonates, cyclic acetals, and ketals. In certain other exemplary embodiments, a nitrogen protecting group is used. Such nitrogen protecting groups include, but are not limited to, carbamates (including methyl, ethyl, and substituted ethyl carbamates (e.g., Troc), to name a few); amides, cyclic imide derivatives, N-alkyl and N-aryl amines, imine derivatives, and enamine derivatives, to name a few. Certain other exemplary protecting groups are detailed herein, however, it is to be understood that the invention is not intended to be limited to these protecting groups; rather, a variety of additional equivalent protecting groups can be readily identified using the criteria described above and used in the present invention. In addition, a variety of protecting Groups are described in "Protective Groups in organic Synthesis" third edition, Greene, T.W. and Wuts, P.G., eds., John Wiley & Sons, New York:1999, the entire contents of which are incorporated herein by reference.

The term "natural amino acid" as used herein refers to any one of the common, naturally occurring L-amino acids found in naturally occurring proteins: glycine (Gly), alanine (Ala), valine (Val), leucine (Leu), isoleucine (Ile), lysine (Lys), arginine (Arg), histidine (His), proline (Pro), serine (Ser), threonine (Thr), phenylalanine (Phe), tyrosine (Tyr), tryptophan (Trp), aspartic acid (Asp), glutamic acid (Glu), asparagine (Asn), glutamine (gin), cysteine (Cys) and methionine (Met).

As used herein, the term "non-natural amino acid" refers to all amino acids that are not natural amino acids, including, for example, α -, β -, D-, L-amino acid residues, and general formulasWherein the side chain R is different from the side chain of a naturally occurring amino acid.

Generally, as used herein, the term "amino acid" includes both natural amino acids and unnatural amino acids.

As used herein, the term "bioisostere" refers broadly to two or more compounds or moieties having similar molecular shape and/or volume. In certain embodiments, the bioisosteres have approximately the same electron distribution. In certain other embodiments, bioisosteres exhibit similar biological properties. In preferred embodiments, bioisosteres have similar molecular shapes and volumes; have approximately the same electron distribution; and exhibit similar biological properties.

As used herein, the term "isolated" when applied to the compounds of the present invention means that such compounds are (i) isolated from at least some of the components with which they are associated in nature or at the time they are produced and/or (ii) produced, prepared, or manufactured by hand.

As used herein, the term "pharmaceutically acceptable derivative" means any pharmaceutically acceptable salt, ester, or salt of such an ester, or any other adduct or derivative of such a compound, which upon administration to a patient is capable of providing (directly or indirectly) a compound or metabolite or residue thereof as described elsewhere herein. Thus, pharmaceutically acceptable derivatives include, inter alia, prodrugs. Prodrugs are derivatives of compounds, which usually have a significantly reduced pharmacological activity, which comprise additional sensitive moieties for removal in vivo to give the parent molecule as the pharmacologically active substance. Examples of prodrugs are esters, which cleave in vivo to give the compound of interest. Prodrugs of a variety of compounds, materials and methods for derivatizing a parent compound to produce a prodrug are known and may be suitable for the present invention. Certain exemplary pharmaceutical compositions and pharmaceutically acceptable derivatives are discussed in more detail herein below.

As used herein, the term "pharmaceutically acceptable salt" refers to those salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without excessive toxicity, irritation, allergic response, and the like, commensurate with a reasonable benefit/risk ratio. Pharmaceutically acceptable salts of amines, carboxylic acids, and other types of compounds are well known to those skilled in the art. Pharmaceutically acceptable salts are described in detail, for example, in J.pharmaceutical Sciences,66:1-19(1977), by S.M.Berge et al, which is incorporated herein by reference. Salts may be prepared in situ during the final isolation and purification of the compounds of the invention, or separately by reacting the free base or free acid functional group with a suitable reagent, as generally described below. For example, the free base functionality can be reacted with a suitable acid. In addition, where the compounds of the invention carry an acidic moiety, suitable pharmaceutically acceptable salts thereof may include metal salts such as alkali metal salts, e.g. sodium or potassium salts; alkaline earth metal salts such as calcium or magnesium salts. Examples of pharmaceutically acceptable, non-toxic acid addition salts are salts of amino groups with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid, or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid, or by other methods used in the art such as ion exchange. Other pharmaceutically acceptable salts include adipates, alginates, ascorbates, aspartates, benzenesulfonates, benzoates, bisulfates, borates, butyrates, camphorates, camphorsulfonates, citrates, cyclopentanepropionates, digluconates, dodecylsulfates, ethanesulfonates, formates, fumarates, glucoheptonates, glycerophosphates, gluconates, hemisulfates, heptanoates, hexanoates, hydroiodides, 2-hydroxyethanesulfonates, lactylates (lactobionates), lactates, laurates, dodecylsulfates, malates, maleates, malonates, methanesulfonates, 2-naphthalenesulfonates, nicotinates, nitrates, oleates, oxalates, palmitates, pamoates, pectinates, persulfates, 3-phenylpropionates, persulfates, salts of citric acid, citrates, and mixtures thereof, Phosphates, picrates, pivalates, propionates, stearates, succinates, sulfates, tartrates, thiocyanates, p-toluenesulfonates, undecanoates, valerates, and the like. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. In addition, pharmaceutically acceptable salts include, where appropriate, non-toxic ammonium salts, quaternary ammonium salts, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, lower alkyl sulfonate and aryl sulfonate.

As used herein, the term "pharmaceutically acceptable ester" refers to esters that hydrolyze in vivo and include those that are easily broken down in the human body to yield the parent compound or a salt thereof. Suitable ester groups include, for example, those derived from pharmaceutically acceptable aliphatic carboxylic acids, particularly alkanoic, alkenoic, naphthenic and alkanedioic acids, in which each alkyl or alkenyl moiety advantageously has not more than 6 carbon atoms. Examples of specific esters include formates, acetates, propionates, butyrates, acrylates, and ethylsuccinates.

As used herein, the term "pharmaceutically acceptable prodrugs" refers to those prodrugs of the compounds of the present invention which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response and the like, commensurate with a reasonable benefit/risk ratio, and effective for their intended use, as well as the zwitterionic forms, where possible, of the compounds of the present invention. The term "prodrug" refers to a compound that is rapidly converted in vivo, e.g., by hydrolysis in blood, to the parent compound of the above formula. A comprehensive discussion is provided in T.Higuchi and V.Stella, Pro-drugs asNovel Delivery Systems, Vol.14of the A.C.S.Symphosis Series and in Edward dB.Roche, ed., Bioreversible Carriers in Drug Design, American pharmaceutical Association and Pergamon Press,1987, both of which are incorporated herein by reference.

As used herein, the term "LFA-1 mediated disease" generally refers to a pathological condition caused by a cell adhesion interaction involving LFA-1 receptors on lymphocytes. Examples of such diseases include, but are not limited to, T cell inflammatory responses such as inflammatory skin diseases, including psoriasis; responses associated with inflammatory bowel disease (such as crohn's disease and ulcerative colitis); adult respiratory distress syndrome; dermatitis; meningitis; encephalitis; uveitis of the eye; allergic conditions such as eczema and asthma; and other conditions involving infiltration of T cells and chronic inflammatory responses; cutaneous hypersensitivity reactions (including poison ivy and poison oak); atherosclerosis; leukocyte adhesion defects; autoimmune diseases such as rheumatoid arthritis, Systemic Lupus Erythematosus (SLE), diabetes, multiple sclerosis, Reynaud's syndrome, autoimmune thyroiditis, experimental autoimmune encephalomyelitis, sjogren's syndrome, type I diabetes, juvenile onset diabetes; and delayed hypersensitivity mediated by cytokines and T lymphocytes typically found in tuberculosis, sarcoidosis, polymyositis, granulomatosis, and vasculitis; pernicious anemia; diseases involving leukocyte extravasation; inflammatory diseases of the CNS, multiple organ damage syndrome secondary to sepsis or trauma, autoimmune hemolytic anemia, myasthenia gravis (myethamia gravis), antigen-antibody complex mediated diseases, and all types of transplantation, including graft versus host disease or host versus graft disease.

As used herein, the term "LFA-1 antagonist" refers broadly to a compound of the invention described herein that acts as a competitive inhibitor of the interaction of CD11a and/or CD18 with ICAM-1, ICAM-2 or ICAM-3.

As used herein, the term "treatment" generally means that the compounds of the present invention are useful in humans or animals, at least for the initial diagnosis of a disease. The compounds of the invention will delay or slow the progression of the disease, thereby extending the life of the individual.

As used herein, the term "prophylaxis" generally means that a compound of the invention is useful for administration to a patient who has not been diagnosed as likely to have the disease at the time of administration but is generally expected to develop into or be at an increasing risk for the disease. In certain embodiments, the compounds of the invention slow the progression of disease symptoms, delay the onset of disease, or completely prevent the development of the disease in a subject.

As used herein, the term "biological sample" includes, but is not limited to, cell cultures or extracts thereof; biopsy material obtained from an animal (e.g., a mammal) or an extract thereof; and blood, saliva, urine, feces, semen, tears, or other bodily fluids, or extracts thereof. For example, the term "biological sample" refers to a sample of any solid or fluid obtained from or excreted or secreted by any biological organism, including unicellular microorganisms (such as bacteria and yeast) and multicellular organisms (such as plants and animals, such as vertebrates or mammals, particularly healthy or apparently healthy human subjects, or human patients affected by a condition or disease being diagnosed or studied). The biological sample can be in any form, including solid materials such as tissues, cells, cell pellets, cell extracts, cell homogenates, or cell fractions; or a biopsy or biological fluid. The biological fluid may be obtained from any location (e.g., blood, saliva (or mouth wash containing buccal cells), tears, plasma, serum, urine, bile, cerebrospinal fluid, amniotic fluid, peritoneal fluid, and pleural fluid, or cells therefrom, aqueous or vitreous humor, or any bodily exudate), exudate, secretions (e.g., fluids from an abscess or any other site of infection or inflammation), or fluids from a joint (e.g., a normal joint or a joint affected by a disease such as rheumatoid arthritis, osteoarthritis, gout, or septic arthritis). The biological sample may be obtained from any organ or tissue (including biopsy or autopsy specimens) or may comprise cells (primary cells or cultured cells) or culture media that has been adapted to any, tissue or organ. Biological samples may also include tissue sections such as frozen sections taken for histological purposes. Biological samples also include mixtures of biomolecules, including proteins, lipids, carbohydrates and nucleic acids, produced by partial or complete fractionation of cells or tissue homogenates. Although it is preferred that the sample is taken from a human subject, the biological sample may be obtained from any animal, plant, bacteria, virus, yeast, etc. As used herein, the term animal refers to humans as well as non-human animals at any stage of development, including, for example, mammals, birds, reptiles, amphibians, fish, worms, and single cells. Cell cultures and biopsy samples are considered to be many animals. In certain exemplary embodiments, the non-human animal is a mammal (e.g., a rodent, a mouse, a rat, a rabbit, a monkey, a dog, a cat, a sheep, a cow, a primate, or a pig). The animal can be a transgenic animal or a human clone. If desired, the biological sample may be subjected to preliminary processing, including preliminary separation techniques.

Detailed description of certain preferred embodiments of the invention

The present invention provides compounds that modulate the interaction between intracellular adhesion molecules (e.g., ICAM-1, -2, and-3) and the leukocyte integrin family of receptors. In certain embodiments, the compounds of the invention are antagonists and are useful in the treatment of CD11/CD18 mediated diseases. In certain embodiments of particular interest, the compounds of the invention are useful for treating Mac-1 and LFA-1 mediated diseases. In other embodiments, the compounds may be used to treat LFA-1 mediated diseases, such as inflammatory and autoimmune diseases, to name a few.

1) General description of the Compounds of the invention

The compounds of the invention include compounds of general formula (I) as further defined below:

and pharmaceutically acceptable derivatives thereof;

wherein R is¹And R²Each independently hydrogen, an amino acid side chain, aliphatic, alicyclic, heteroaliphatic, heterocyclic, aromatic or heteroaromatic moiety, or

Wherein R is¹And R²Together being a cycloaliphatic or heterocyclic moiety, or together being

Wherein R is^1AIs hydrogen, aliphatic, alicyclic, heteroaliphatic, heterocyclic, aromatic, or heteroaromatic moiety;

R³is-C (═ O) OR^3A、-C(＝O)H、-CH₂OR^3A、-CH₂O-C (═ O) -alkyl, -C (═ O) NH (R)^3A) or-CH₂X⁰Wherein R is^3AIndependently at each occurrence is hydrogen, a protecting group, aliphatic, alicyclic, heteroaliphatic, heterocyclic, aromatic, or heteroaromatic moiety, or R^3AAnd R¹Or R²Together form a heterocyclic moiety wherein X⁰Is halogen selected from F, Cl, Br or I;

R⁴independently at each occurrence is hydrogen, halogen, -CN, -NO₂Aliphatic, alicyclic, heteroaliphatic, heterocyclic, aromatic, or heteroaromatic moiety, or is-GR^G1Wherein G is-O-, -S-, -NR^G2-、-CO-、-SO-、-SO₂-、-C(＝O)O-、-C(＝O)NR^G2-、-OC(＝O)-、-NR^G2C (═ O) -or-SO)₂NR^G2-，R^G1And R^G2Independently hydrogen, aliphatic, alicyclic, heteroaliphatic, heterocyclic, aromatic, or heteroaromatic moiety;

n is an integer of 0 to 3;

AR¹is an aromatic, heteroaromatic, alicyclic, or hetero moietyA ring portion;

A. b, D and E are connected by a single or double bond, where valency permits; wherein A, B, D and E are independently at each occurrence C-O, CRⁱRⁱⁱ、NRⁱ、CRⁱN, O, S, S (═ O) or SO₂(ii) a Wherein R isⁱIndependently at each occurrence is hydrogen, halogen, -CN, -NO₂Aliphatic, alicyclic, heteroaliphatic, heterocyclic, aromatic, or heteroaromatic moiety, or is-GR^G1Wherein G is-O-, -S-, -NR^G2-、-CO-、-SO-、-SO₂-、-C(＝O)O-、-C(＝O)NR^G2-、-OC(＝O)-、-NR^G2C (═ O) -or-SO)₂NR^G2-，R^G1And R^G2Independently hydrogen, aliphatic, alicyclic, heteroaliphatic, heterocyclic, aromatic, or heteroaromatic moiety, or RⁱAny two adjacent occurrences of (a) together represent an alicyclic, heterocyclic, aromatic, or heteroaromatic moiety;

p is an integer of 0 to 4; and

l is absent or is V-W-X-Y-Z, wherein V, W, X, Y and Z are independently absent at each occurrence, C-O, NR^L1、-O-、-C(R^L1)＝、＝C(R^L1)-、-C(R^L1)(R^L2)、C(＝N-O-R^L1)、C(＝N-R^L1)、-N＝、S(O)_0-2(ii) a Substituted or unsubstituted C_1-6Alkylene or C_2-6Alkenylene chains in which up to two non-adjacent methylene units are independently optionally substituted by-C (═ O) -, -CO₂-、-C(＝O)C(＝O)-、-C(＝O)NR^L3-、-OC(＝O)-、-OC(＝O)NR^L3-、-NR^L3NR^L4-、-NR^L3NR^L4C(＝O)-、-NR^L3C(＝O)-、-NR^L3CO₂-、-NR^L3C(＝O)NR^L4-、-S(＝O)-、-SO₂-、-NR^L3SO₂-、-SO₂NR^L3-、-NR^L3SO₂NR^L4-, -O-, -S-, or-NR^L3-substitution; wherein R is^L3And R^L4Independently at each occurrence is hydrogen, alkyl, heteroalkyl, aromatic moiety, heteroaromatic moiety, or acyl; or an aliphatic, alicyclic, heteroaliphatic, heterocyclic, aromatic, or heteroaromatic moiety; and R is^L1And R^L2Independently at each occurrence is hydrogen, hydroxy, protected hydroxy, amino, protected amino, thio, protected thio, halogen, cyano, isocyanate, carboxy, carboxyalkyl, formyl, formyloxy, azido, nitro, ureido, thioureido, thiocyanato, alkoxy, aryloxy, mercapto, sulfonamido, benzamido, tosyl, or an aliphatic, cycloaliphatic, heteroaliphatic, heterocyclic, aromatic, or heteroaromatic moiety, or wherein R occurs one or more times^L1And R^L2Together, or together with V, W, X, Y or one of Z, form an alicyclic or heterocyclic moiety or form an aromatic or heteroaromatic moiety.

In another aspect, the invention provides a compound of formula (II):

wherein AR has one of the following structures:

and pharmaceutically acceptable derivatives thereof;

wherein R is¹、R²、R³、R⁴A, B, D, E, n, p are generally as defined above and in classes and subclasses herein; and

Y¹、Y²and Y³Each independently is CR⁴Or N;

provided that when AR has the following structure

Wherein Y is¹Is CH or N, and p is 0-2,

then R is⁴Is not a carbocyclic, aryl, heteroaryl, or heterocyclic ring, and A, B, D and E do not include carbocyclic moieties, aryl, heteroaryl, or heterocyclic moieties.

In certain embodiments, for compounds of formula (II), AR represents a moiety having one of the following structures:

wherein each occurrence of n is an integer from 0 to 6; r⁴Independently at each occurrence is hydrogen, halogen, CN, isocyanate, NO₂、-P(＝O)(YR^P5)₂Alkyl, cycloalkyl, heteroalkyl, heterocyclic moiety, or is-GR^G1Wherein G is-O-, -S-, -NR^G2-、-CO-、-SO-、-SO₂-、-C(＝O)O-、-C(＝O)NR^G2-、-OC(＝O)-、-NR^G2C (═ O) -or-SO)₂NR^G2-, and R^G1And R^G2Independently hydrogen, alkyl, cycloalkyl, heteroalkyl, heterocyclic moieties. Y is independently at each occurrence a bond or O; r^P5Independently at each occurrence is alkyl, heteroalkyl, aryl or heteroaryl, or when Y is O, R^P5May also be hydrogen, and R^4AIndependently at each occurrence is hydrogen, alkyl, cycloalkyl, heteroalkyl, a heterocyclic moiety or a nitrogen protecting group; wherein R is⁴And R^4AAny two adjacent occurrences of (a) together can form a cycloalkyl, heterocyclic, aryl, or heteroaryl group. In some exemplary embodimentsIn one embodiment, the AR has the following structure:

in other exemplary embodiments, the AR has the following structure:wherein X₀Independently at each occurrence is a halogen selected from F, Cl, Br and I. In certain embodiments, X⁰At each occurrence is Cl.

A number of important sub-classes of each of the aforementioned classes deserve separate mention, including sub-classes of the aforementioned classes, wherein:

i)R¹and R²Each independently hydrogen, an amino acid side chain, - (CH)₂)_mOH、-(CH₂)_mAryl, - (CH)₂)_mHeteroaryl (wherein m is 0-6), -CH (R)^1A)(OR^1B)、-CH(R^1A)(NHR^1B) U-T-Q, or an alkyl, cycloalkyl, heteroalkyl, or heterocyclic moiety optionally substituted with U-T-Q, wherein U is absent, -O-, -S (O)_0-2-、-SO₂N(R^1A)、-N(R^1A)-、-N(R^1A)C(＝O)-、-N(R^1A)C(＝O)-O-、-N(R^1A)C(＝O)-N(R^1B)-、-N(R^1A)-SO₂-, -C (═ O) -O-, -O-C (═ O) -, aryl, heteroaryl, alkylaryl, alkylheteroaryl, -C (═ O) -N (R)^1A)-、-O-C(＝O)-N(R^1A)-、-C(＝N-R^1E)-、-C(＝N-R^1E)-O-、-C(＝N-R^1E)-N(R^1A)-、-O-C(＝N-R^1E)-N(R^1A)-、-N(R^1A)C(＝N-R^1E)-、-N(R^1A)C(＝N-R^1E)-O-、N(R^1A)C(＝N-R^1E)-N(R^1B)-、-P(＝O)(OR^1A) -O-, or-P (═ O) (R)^1A) -O-; wherein T is absent, alkyl, cycloalkyl, heteroalkyl, heterocyclic, aryl, heteroaryl, alkylaryl, alkylheteroaryl, heteroalkylAn aryl OR heteroalkyl heteroaryl moiety, and wherein Q is hydrogen, halogen, cyano, isocyanate, -OR^1B、-SR^1B；-N(R^1B)₂、-NHC(＝O)OR^1B、-NHC(＝O)N(R^1B)₂、-NHC(＝O)R^1B、-NHSO₂R^1B、-NHSO₂N(R^1B)₂、-NHSO₂NHC(＝O)OR^1B、-NHC(＝O)NHSO₂R^1B、-C(＝O)NHC(＝O)OR^1B、-C(＝O)NHC(＝O)R^1B、-C(＝O)NHC(＝O)N(R^1B)₂、-C(＝O)NHSO₂R^1B、-C(＝O)NHSO₂N(R^1B)₂、-C(＝S)N(R^1B)₂、-SO₂R^1B、-SO₂-O-R^1B、-SO₂-N(R^1B)₂、-SO₂-NHC(＝O)OR^1B、-SO₂-NHC(＝O)-N(R^1B)₂、-SO₂-NHC(＝O)R^1B、-O-C(＝O)N(R^1B)₂、-O-C(＝O)R^1B、-O-C(＝O)NHC(＝O)R^1B、-O-C(＝O)NH-SO₂R^1B、-O-SO₂R^1BOr an alkyl, cycloalkyl, heteroalkyl, heterocyclic, aryl or heteroaryl moiety, or wherein R is¹And R²Together are a cycloalkyl heterocyclic moiety, or together areWherein R is^1AAnd R^1BIndependently at each occurrence is hydrogen, alkyl, cycloalkyl, heteroalkyl, heterocyclic, aryl or heteroaryl moiety, -COR^1Cor-CONR^1CR^1D(ii) a Wherein R is^1CAnd R^1DIndependently at each occurrence is hydrogen, hydroxy, or an alkyl, cycloalkyl, heteroalkyl, heterocyclic, aryl or heteroaryl moiety; and R^1EIs hydrogen, an aliphatic, alicyclic, heteroaliphatic moiety, heterocyclic, aryl, heteroaryl, alkylaryl OR alkylheteroaryl moiety, -CN, -OR^1C、-NR^1CR^1Dor-SO₂R^1C；

ii)R³Is carboxy, protected carboxy or a prodrug thereof, wherein R³Is C (═ O) R^3AWherein R is^3AIs hydroxy, alkoxy, cycloalkoxy, aralkoxy, arylcycloalkoxy, aryloxy, alkylcarbonyloxyalkyloxy, alkoxycarbonyloxyalkyloxy, alkoxycarbonylalkyloxy, cycloalkylcarbonyloxyalkyloxy, cycloalkoxycarbonyloxyalkyloxy, cycloalkoxycarbonylalkyloxy, arylcarbonyloxyalkyloxy, aryloxycarbonyloxyalkyloxy, arylcarbonyloxyalkyloxy, alkoxyalkylcarbonyloxyalkyloxy, or one of the following structures:

iii)R³is-C (═ O) OR^3A、-C(＝O)H、-CH₂OR^3A、-CH₂O-C (═ O) -alkyl, -C (═ O) NH (R)^3A) or-CH₂X⁰(ii) a Wherein R is^3AIndependently at each occurrence is hydrogen, a protecting group, an alkyl, cycloalkyl, heteroalkyl, heterocyclic, aryl, heteroaryl, alkylaryl or alkylheteroaryl moiety, or R^3AAnd R¹Or R²Together form a heterocyclic moiety; wherein X⁰Is halogen selected from F, Cl, Br or I;

iv)R³is-C (═ O) OR^3A(ii) a Wherein R is^3AIs hydrogen, a protecting group, an alkyl, cycloalkyl, heteroalkyl, heterocyclic, aryl, heteroaryl, alkylaryl or alkylheteroaryl moiety, or R^3AAnd R¹Or R²Together form a heterocyclic moiety;

v)R³is-C (═ O) OR^3A(ii) a Wherein R is^3AIs C_1-5An alkyl group;

vi)R³is-C (═ O) OR^3A(ii) a Wherein R is^3AIs C_1-3An alkyl group;

vii)R³is-C (═ O) OR^3A(ii) a Wherein R is^3AIs ethyl;

viii)R³is-C (═ O) OR^3A(ii) a Wherein R is^3AIs benzyl;

ix)R³is CO₂H；

x)R³is-C (═ O) OR^3AWherein R is^3AAs defined in any of the above subsets ii) -ix), -C (═ O) NHC (R)¹)(R²)R³Is a moiety having the structure:

wherein Ar is₂Is a cycloalkyl, heterocyclic, aryl or heteroaryl moiety; and R^SIs hydrogen, alkyl, heteroalkyl, aryl, heteroaryl, or is-G⁰R^G1Wherein G is⁰is-O-, -S-or-NR^G2-，R^G1And R^G2Independently hydrogen, aliphatic, alicyclic, heteroaliphatic, heterocyclic, aromatic, or heteroaromatic moiety;

xi) wherein-C (═ O) NHCH (CO)₂R^3A)CH(R^S)Ar₂Compounds of the above subset x) having the following stereochemistry:

xii)R³is-C (═ O) OR^3AWherein R is^3AAs defined in any one of the above subsets ii) -ix), and-C (═ O) NHC (R)¹)(R²)R³Is a moiety having the structure:

wherein R is^1AIs Ar₂、-OR^1B、-SR^1Bor-NR^1BR^1C(ii) a Or an alkyl or heteroalkyl moiety; ar (Ar)₂Is a cycloalkyl, heterocyclic, aryl or heteroaryl moiety; wherein R is^1BAnd R^1CIndependently hydrogen, alkyl, heteroalkyl, cycloalkyl, heterocyclic, aryl, heteroaryl, or R^1BAnd R^1CTogether with the nitrogen atom to which they are attached form a heterocyclic or heteroaryl moiety;

xiii) wherein-C (═ O) NHCH (CO)₂R^3A)CH₂NHC(＝O)R^1ACompounds of the above subset xii) having the following stereochemistry:

xiv)R³is-C (═ O) OR^3AWherein R is^3AAs defined in any one of the above subsets ii) -ix), and-C (═ O) NHC (R)¹)(R²)R³Is a moiety having the structure:

wherein Ar is₂Is a cycloalkyl, heterocyclic, aryl or heteroaryl moiety; r^2AIs hydrogen, C_1-6Alkyl radical, C_2-6Alkenyl, -C (═ O) R^2Bor-SO₂R^2BWherein R is^2BIs alkyl, cycloalkyl, heteroalkyl, heterocyclyl, aryl or heteroaryl; or R^2AAnd Ar₂The substituents on (a) together form a substituted or unsubstituted heterocyclic or heteroaryl moiety;

xv) wherein-C (═ O) NHCH (CO)₂R^3A)CH₂N(R^2A)Ar₂Compounds of the above subset xiv) having the following stereochemistry:

xvi)R³is-C (═ O) OR^3AWherein R is^3AAs defined in any one of the above subsets ii) -ix), and-C (═ O) NHC (R)¹)(R²)R³Is a moiety having the structure:

wherein R is^2AIs hydrogen, C_1-6Alkyl radical, C_2-6Alkenyl, aryl, heteroaryl, -C (═ O) R^2Bor-SO₂R^2BWherein R is^2BIs alkyl, cycloalkyl, heteroalkyl, heterocyclyl, aryl or heteroaryl; or R^2AAnd R^2COr R^2DTogether form a substituted or unsubstituted heterocyclic or heteroaryl moiety; r^2CIs hydrogen, CN, -C ═ NMe, or NO₂、＝NC(＝O)NH₂、＝NS(O)₂R、＝NS(O)₂NRR′、-SO₂R^2GOr an aliphatic, alicyclic, heteroaliphatic, heteroalicyclic, aryl, heteroaryl, alkylaryl or alkylheteroaryl moiety; wherein R and R' are each independently hydrogen or methyl, R^2GIs lower alkyl; r^2DIs Ar₂Hydrogen, halogen, CN, NO₂Aliphatic, heteroaliphatic, alkylaryl or alkylheteroaryl moiety, or is-GR^G1Wherein G is-O-, -S-, -NR^G2-、-CO-、-SO-、-SO₂-、-C(＝O)O-、-C(＝O)NR^G2-、-OC(＝O)-、-NR^G2C (═ O) -or-SO)₂NR^G2-，R^G1And R^G2Independently is a hydrogen, aliphatic, alicyclic, heteroaliphatic, heteroalicyclic, aryl, heteroaryl, alkylaryl, or alkylheteroaryl moiety;

xvii) wherein-C (═ O)NHCH(CO₂R^3A)CH₂N(R^2A)C(＝NR^2C)R^2DCompounds of the above subset xvi) having the following stereochemistry:

xviii) wherein-C (═ O) NHCH (CO)₂R^3A)CH₂N(R^2A)C(＝NR^2C)R^2DCompounds of the above subset xvii) having the following structure:

wherein R is^2EAnd R^2FEach independently hydrogen, or an aliphatic, alicyclic, heteroaliphatic, heteroalicyclic, aryl, heteroaryl, alkylaryl or alkylheteroaryl moiety, or R^2EAnd R^2FTogether form a substituted or unsubstituted heterocyclic or heteroaryl moiety;

xix) wherein-C (═ O) NHCH (CO)₂R^3A)CH₂N(R^2A)C(＝NR^2C)R^2DCompounds of the above subset xvii) having the following structure:

wherein R is^2CIs hydrogen, CN, -C ═ NMe, or NO₂、＝NC(＝O)NH₂、＝NS(O)₂R or NS (O)₂NRR'; wherein R and R' are each independently hydrogen or methyl;

xx) wherein-C (═ O) NHCH (CO)₂R^3A)CH₂N(R^2A)C(＝NR^2C)R^2DCompounds of the above subset xvii) having the following structure:

wherein R is^2CIs hydrogen, CN, -C ═ NMe, or NO₂、＝NC(＝O)NH₂、＝NS(O)₂R or NS (O)₂NRR'; wherein R and R' are each independently hydrogen or methyl;

xxi) wherein-C (═ O) NHCH (CO)₂R^3A)CH₂N(R^2A)C(＝NR^2C)R^2DCompounds of the above subset xvii) having the following structure:

xxii) wherein-C (═ O) NHCH (CO)₂R^3A)CH₂N(R^2A)C(＝NR^2C)R^2DCompounds of the above subset xvii) having the following structure:

xxiii) wherein-C (═ O) NHCH (CO)₂R^3A)CH₂N(R^2A)C(＝NR^2C)R^2DCompounds of the above subsets xvii) and xviii) having the following structures:

or a bioisostere thereof;

wherein R is^2A、R^2D、R^2EAnd R^2FAs defined in xvi) and xviii) above;

xxiv) compounds in which the bioisostere has a subset xxiii) of one of the following structures:

wherein R is^2CIs lower alkyl;

xxv) wherein R^2DIs or R^2EAnd R^2FCompounds of the above subset xxiii) which together with the nitrogen atom to which they are attached form part of one of the following structures:

wherein s is an integer from 0 to 6; r^P1Independently at each occurrence is hydrogen, halogen, CN, isocyanate, NO₂、-P(＝O)(YR^P5)₂An alkyl, cycloalkyl, heteroalkyl, heterocyclic, aryl, heteroaryl, alkylaryl or alkylheteroaryl moiety, or is-GR^G1Wherein G is-O-, -S-, -NR^G2-、-CO-、-SO-、-SO₂-、-C(＝O)O-、-C(＝O)NR^G2-、-OC(＝O)-、-NR^G2C (═ O) -or-SO)₂NR^G2-，R^G1And R^G2Independently is a hydrogen, alkyl, cycloalkyl, heteroalkyl, heterocyclic, aryl, heteroaryl, alkylaryl or alkylheteroaryl moiety; y is independently at each occurrence a bond or O; r^P5Independently at each occurrence, is alkyl, heteroalkyl, aryl or heteroaryl, or R is O when Y is O^P5May also be hydrogen; r^P2Independently at each occurrence is hydrogen, an aliphatic, alicyclic, heteroaliphatic, heterocyclic, aryl, heteroaryl, alkylaryl, alkylheteroaryl, heteroalkylaryl, or heteroalkylheteroaryl moiety or a nitrogen protecting group; any two of which are adjacentR^P1And R^P2Together may form a cycloalkyl, heterocyclic, aryl or heteroaryl moiety;

xxvi) wherein R^2DIs or R^2EAnd R^2FCompounds of the above subset xxv) which together with the nitrogen atom to which they are attached form part of one of the following structures:

wherein R is^P1Independently at each occurrence is hydrogen, halogen, methyl, -OCH₃、-OH、-NH₂、-NHCH₃or-N (CH)₃)₂；

xxvii) wherein R^2DIs or R^2EAnd R^2FCompounds of the above subset xxvi) which together with the nitrogen atom to which they are attached form part of one of the following structures:

xxviii)R³is-C (═ O) OR^3AWherein R is^3A-C (═ O) NHC (R) as defined in any one of the above subsets ii) -ix)¹)(R²)R³Is a moiety having the structure:

wherein Ar is₂Is a cycloalkyl, heterocyclic, aryl or heteroaryl moiety;

xxix) compounds of sub-sets x) -xii), xiv) -xv) and xxviii); and wherein R^2DIs Ar₂(ii) a Wherein Ar is₂A compound that is a subset xvi) of one of the following structures:

wherein s is an integer from 0 to 6 at each occurrence; w is an integer of 0 to 6; x₁Is CHR^P1Or NR^P2；X₂And X₃Independently CHR^P1、NR^P2、CHSO₂R^P3Or NSO₂R^P3；R^P1Independently at each occurrence is hydrogen, halogen, CN, isocyanate, NO₂、-P(＝O)(YR^P5)₂Aliphatic, alicyclic, heteroaliphatic, heterocyclic, aryl, heteroaryl, - (aliphatic) aryl or- (aliphatic) heteroaryl moieties, or is-GR^G1Wherein G is-O-, -S-, -NR^G2-、-CO-、-SO-、-SO₂-、-C(＝O)O-、-C(＝O)NR^G2-、-OC(＝O)-、-NR^G2C (═ O) -or-SO)₂NR^G2-，R^G1And R^G2Independently is a hydrogen, aliphatic, alicyclic, heteroaliphatic, heterocyclic, aryl, heteroaryl, - (aliphatic) aryl or- (aliphatic) heteroaryl moiety; y is independently at each occurrence a bond or O; r^P5Independently at each occurrence, is alkyl, heteroalkyl, aryl or heteroaryl, or R is O when Y is O^P5May also be hydrogen; r^P2Independently at each occurrence is hydrogen, an aliphatic, alicyclic, heteroaliphatic, heterocyclic, aryl, heteroaryl, alkylaryl, alkylheteroaryl, heteroalkylaryl, or heteroalkylheteroaryl moiety or a nitrogen protecting group; wherein any two adjacent R^P1And R^P2Together may form a cycloalkyl, heterocyclic, aryl or heteroaryl moiety; r^P3Independently at each occurrence isAlkyl, aryl, heteroaryl or-N (R)^P2)₂。

xxx) wherein Ar₂Compounds of the above subset xxix) which are one of the following structures:

wherein s, X¹、X²And X³As defined above in xx); x⁵Is O, S or NR^P2；R^P1Independently at each occurrence is hydrogen, halogen, CN, isocyanate, NO₂、-P(＝O)(YR^P5)₂An alkyl, cycloalkyl, heteroalkyl, heterocyclic, aryl, heteroaryl, alkylaryl or alkylheteroaryl moiety, or is-GR^G1Wherein G is-O-, -S-, -NR^G2-、-CO-、-SO-、-SO₂-、-C(＝O)O-、-C(＝O)NR^G2-、-OC(＝O)-、-NR^G2C (═ O) -or-SO)₂NR^G2-，R^G1And R^G2Independently is a hydrogen, alkyl, cycloalkyl, heteroalkyl, heterocyclic, aryl, heteroaryl, alkylaryl or alkylheteroaryl moiety; y is independently at each occurrence a bond or O; r^P5Independently at each occurrence is alkyl, heteroalkyl, aryl or heteroaryl, or R when Y is O^P5May also be hydrogen; r^P2Independently at each occurrence is hydrogen, an aliphatic, alicyclic, heteroaliphatic, heterocyclic, aryl, heteroaryl, alkylaryl, alkylheteroaryl, heteroalkylaryl, or heteroalkylheteroaryl moiety or a nitrogen protecting group; wherein any two adjacent R^P1And R^P2Together may form a cycloalkyl, heterocyclic, aryl or heteroaryl moiety; r^P3Independently at each occurrence is alkyl, aryl, heteroaryl or-N (R)^P2)₂；

xxxi) Compounds of the above sub-set xxx), in which R^P1Independently at each occurrence, is hydrogen, halogen, -P (═ O) (YR)^P5)₂Lower alkyl or heteroalkyl moiety, or–GR^G1Wherein G is-O-, -S-, -NR^G2-or-SO₂-，R^G1And R^G2Independently hydrogen, lower alkyl or aryl; y is independently at each occurrence a bond or O; r^P5Independently at each occurrence, is lower alkyl, or R when Y is O^P5May also be hydrogen; r^P2Independently at each occurrence is hydrogen, lower alkyl or a nitrogen protecting group; wherein any two adjacent R^P1And R^P2Together may form a cycloalkyl, heterocyclic, aryl or heteroaryl moiety;

xxxii) wherein Ar₂Compounds of the above subset xxx) having one of the following structures:

wherein X₁Is N or CR^P1(ii) a s is an integer of 0 to 6; r^P1Independently at each occurrence is hydrogen, halogen, CN, NO₂An alkyl, cycloalkyl, heteroalkyl, heterocyclic, aryl, heteroaryl, alkylaryl or alkylheteroaryl moiety, or is-GR^G1Wherein G is-O-, -S-, -NR^G2-、-CO-、-SO-、-SO₂-、-C(＝O)O-、-C(＝O)NR^G2-、-OC(＝O)-、-NR^G2C (═ O) -or-SO)₂NR^G2-，R^G1And R^G2Independently is a hydrogen, alkyl, cycloalkyl, heteroalkyl, heterocyclic, aryl, heteroaryl, alkylaryl or alkylheteroaryl moiety; r^P3Independently is lower alkyl or aryl;

xxxiii) compounds of the sub-sets xxix), xxx) and xxxii) wherein s is 0;

xxxiv) xxix), xxx), and xxxii) compounds of the subset wherein s is 1;

xxxv) compounds of the sub-set xxix) where s is 2, xxx) and xxxii);

xxxvi) wherein Ar₂The above subsets x) and xi) of one of the following structuresCompound (a):

wherein s is an integer of 0 to 2; r^P1Independently at each occurrence is hydrogen, halogen, CN, isocyanate, NO₂、–OR^G1、-SR^G1、-NR^G1R^G2-, alkyl, cycloalkyl, heteroalkyl, heterocyclic, aryl, heteroaryl, alkylaryl or alkylheteroaryl moieties; y is independently at each occurrence a bond or O; r^P5Independently at each occurrence, is lower alkyl, or R when Y is O^P5May also be hydrogen; r^P2Independently at each occurrence is hydrogen, alkyl, cycloalkyl, heteroalkyl, heterocyclic, aryl, heteroaryl, alkylaryl, alkylheteroaryl, heteroalkylaryl, or heteroalkylheteroaryl moiety or a nitrogen protecting group; r^P3Is lower alkyl or-N (R)^P2)₂；R^G1And R^G2Independently is a hydrogen, alkyl, cycloalkyl, heteroalkyl, heterocyclic, aryl, heteroaryl, alkylaryl or alkylheteroaryl moiety;

xxxvii) wherein Ar₂Compounds of the above subsets x) and xi) being one of the following structures:

xxxviii) wherein Ar₂Compounds of the above subsets x) and xi) being one of the following structures:

wherein R is^P3Is lower alkyl; r^P2And R^G1Independently hydrogen or lower alkyl;

xxxix) wherein Ar₂Is a lower knotCompounds of the above subsets x) and xi) of one of the structures:

wherein R is^P3Is lower alkyl, R^G1Is hydrogen or lower alkyl;

xl) wherein R^SIs hydrogen, hydroxy or lower alkoxy and Ar₂Compounds of the above subsets x) and xi) being one of the following structures:

wherein R is^P3Is lower alkyl; r^G1Is hydrogen or lower alkyl;

xli) wherein R^1AIs alkyl or-NR^1BR^1CCompounds of sub-sets xii) and xiii); wherein R is^1BAnd R^1CIndependently hydrogen or lower alkyl;

xlii) wherein R^1Ais-NH₂Or a subset xii) and xiii) of the moieties having the following structures:

wherein R is^P1Independently hydrogen, hydroxy, lower alkyl or lower heteroalkyl; r^P2Independently at each occurrence is hydrogen or lower alkyl;

xliii) wherein R^1Ais-NH₂Or a subset xii) and xiii) of the moieties having the following structures:

wherein R is^P1Is hydrogen or lower alkyl;

xliv) wherein R^1ACompounds that are a subset xii) and xiii) of cycloalkyl, aryl, or a moiety having one of the following structures:

wherein s is an integer from 0 to 6; r^P1Independently at each occurrence is hydrogen, halogen, CN, isocyanate, NO₂、-P(＝O)(YR^P5)₂An alkyl, cycloalkyl, heteroalkyl, heterocyclic, aryl, heteroaryl, alkylaryl or alkylheteroaryl moiety, or is-GR^G1Wherein G is-O-, -S-, -NR^G2-、-CO-、-SO-、-SO₂-、-C(＝O)O-、-C(＝O)NR^G2-、-OC(＝O)-、-NR^G2C (═ O) -or-SO)₂NR^G2-，R^G1And R^G2Independently is a hydrogen, alkyl, cycloalkyl, heteroalkyl, heterocyclic, aryl, heteroaryl, alkylaryl or alkylheteroaryl moiety; y is independently at each occurrence a bond or O; r^P5Independently at each occurrence is alkyl, heteroalkyl, aryl or heteroaryl, or R when Y is O^P5May also be hydrogen; r^P2Independently at each occurrence is hydrogen, an aliphatic, alicyclic, heteroaliphatic, heterocyclic, aryl, heteroaryl, alkylaryl, alkylheteroaryl, heteroalkylaryl, or heteroalkylheteroaryl moiety or a nitrogen protecting group; wherein any two adjacent R^P1And R^P2Together may form a cycloalkyl, heterocyclic, aryl or heteroaryl moiety;

xlv) a sub-set xliv), wherein s is an integer from 0 to 2; r^P1Independently at each occurrence, is lower alkyl or is-GR^G1Wherein G is-O-or-NR^G2-，R^G1And R^G2Independently is a hydrogen, alkyl, cycloalkyl, heteroalkyl, heterocyclic, aryl, heteroaryl, alkylaryl or alkylheteroaryl moietyDividing; r^P2Independently at each occurrence is hydrogen, lower alkyl, aryl or heteroaryl;

xlvi) wherein R^1ACompounds which are a subset of moieties having one of the following structures xxi) and xxii):

wherein s is an integer from 0 to 2; x⁰Is halogen; r^P1Independently at each occurrence is hydrogen, hydroxy, lower alkyl or lower heteroalkyl; g is-O-or-NR^G2-，R^G1And R^G2Independently hydrogen or lower alkyl; r^P2Independently hydrogen or lower alkyl;

xlvii) wherein R^1AA compound of the xlvi) subset which is part of one of the following structures:

wherein G is-O-or-NR^G2-，R^G1And R^G2Independently hydrogen or lower alkyl;

xlviii)R³is-C (═ O) OR^3AWherein R is^3A-C (═ O) NHC (R) as defined in any one of the above subsets ii) -ix)¹)(R²)R³Is a moiety having the structure:

wherein R is^P3Is an alkyl, cycloalkyl, heterocyclic, aryl or heteroaryl moiety;

xlix) wherein-NH (R)^2A)Ar₂Compounds of the above subset xiv) -xv) having one of the following structures:

wherein X₁Is N or CR^P1(ii) a s is an integer of 0 to 5; r^P1Independently at each occurrence is hydrogen, halogen, CN, NO₂An alkyl, cycloalkyl, heteroalkyl, heterocyclic, aryl, heteroaryl, alkylaryl or alkylheteroaryl moiety, or is-GR^G1Wherein G is-O-, -S-, -NR^G2-、-CO-、-SO-、-SO₂-、-C(＝O)O-、-C(＝O)NR^G2-、-OC(＝O)-、-NR^G2C (═ O) -or-SO)₂NR^G2-，R^G1And R^G2Independently is a hydrogen, alkyl, cycloalkyl, heteroalkyl, heterocyclic, aryl, heteroaryl, alkylaryl or alkylheteroaryl moiety; and R^P3Is alkyl, heteroalkyl, aryl or heteroaryl;

l) compounds of the above-mentioned subset xlix) in which s is 0;

li) wherein R^P1Compounds of the above sub-group xlix) which are hydrogen, halogen or lower alkyl;

lii) wherein R^P1Compounds of the above sub-group li) which are hydrogen, chloro or methyl;

liii) wherein R^P3Compounds of the above sub-group xlix) which are lower alkyl;

liv) wherein R^P3Compounds of the above subset liii) which are methyl;

lv) wherein-NH (R)^2A)Ar₂Compounds of the above subset xlix) having the following structure:

wherein R is^P1Is hydrogen, halogen or lower alkyl;

lvi) wherein-NH (R)^2A)Ar₂Compounds of the above subset xlix) having the following structure:

lvii) compounds having the following structure subset xvii):

or a bioisostere thereof;

wherein R is^P1Independently at each occurrence is hydrogen, halogen, methyl, -OCH₃、-OH、-NH₂、-NHCH₃or-N (CH)₃)₂；R^2AIs hydrogen, C_1-6Alkyl radical, C_2-6Alkenyl, aryl, heteroaryl, -C (═ O) R^2Bor-SO₂R^2BWherein R is^2BIs alkyl, cycloalkyl, heteroalkyl, heterocyclyl, aryl or heteroaryl; and q is 1 or 2;

lviii) compounds of the above subset lvii) wherein the bioisostere has one of the following structures:

wherein q is 1 or 2; and R^2CIs lower alkyl;

lix) wherein-C (═ O) NHC (═ CHAr)₂)CO₂R^3ACompounds having the sub-set xxviii) of one of the following structures:

whereinR^P3Is lower alkyl or aryl; x₁And X₂Independently is N or CR^P1；X₃Is O, S or NR^P2(ii) a Wherein R is^P1Is hydrogen, halogen, CN, NO₂An alkyl, cycloalkyl, heteroalkyl, heterocyclic, aryl, heteroaryl, alkylaryl or alkylheteroaryl moiety, or is-GR^G1Wherein G is-O-, -S-, -NR^G2-、-CO-、-SO-、-SO₂-、-C(＝O)O-、-C(＝O)NR^G2-、-OC(＝O)-、-NR^G2C (═ O) -or-SO)₂NR^G2-, and R^G1And R^G2Independently is a hydrogen, alkyl, cycloalkyl, heteroalkyl, heterocyclic, aryl, heteroaryl, alkylaryl or alkylheteroaryl moiety; and R^P2Is a hydrogen, aliphatic, alicyclic, heteroaliphatic, heterocyclic, aryl, heteroaryl, alkylaryl, alkylheteroaryl, heteroalkylaryl, or heteroalkylheteroaryl moiety;

lx) wherein-C (═ O) NHC (═ CHAr)₂)CO₂R^3ACompounds having the sub-group xxviii) of the structure

Wherein X₁Is N or CH;

lxi)R³is-C (═ O) OR^3AWherein R is^3AAs defined in any one of the above subsets ii) -ix), and-C (═ O) NHC (R)¹)(R²)R³Is the structure-C (═ O) NHC (═ C (R)^S)Ar₂)CO₂R^3AWherein R is^3AAnd R^STogether form a substituted or unsubstituted heterocyclic moiety;

lxii) wherein-C (═ O) NHC (═ C (R)^S)Ar₂)CO₂R^3ACompounds having a subset lxi of one of the following structures):

wherein Ar is₂As defined in classes and subclasses herein; and X₁O, S or NH;

ixiii) wherein-C (═ O) NHC (═ C (R)^S)Ar₂)CO₂R^3ACompounds having a subset lxi of one of the following structures):

wherein X₁O, S or NH; and X₂Is N or CH;

lxxv) L is absent, -C (═ O), -CH₂C(＝O)NH-、-CH₂NH-C(＝O)-、-O-CH₂-C(＝O)-、-CH₂-CH₂-C(＝O)-、-CH＝CH-C(＝O)NH-CH₂-、-CH(OH)-CH₂-O-、-CH(OH)-CH₂-N(CH₃)-、-CH(OH)-CH₂-CH₂-、-CH₂-CH₂-CH(OH)-、-O-CH₂-CH(OH)-、-O-CH₂-CH(OH)-CH₂-、-O-CH₂-CH₂-CH(OH)-、O-CH₂-CH₂-O-、-CH₂-CH₂-CH₂-O-、-CH₂-CH(OH)-CH₂-O、-CH₂-CH₂-O-、-CH-(CH₃)-NH-C(＝O)-、-CH₂-NH-SO₂-、-NH-SO₂-CH₂-、-CH₂-SO₂-NH-、-SO₂NH-CH₂-、-C(＝O)-NH-C(＝O)-、-NH-C(＝O)-NH-、-NH-C(＝O)-NH-CH₂-、-CH₂-NH-C(＝O)-NH-、-C(＝O)-NH-CH₂-C (═ O) -NH, -NH-C (═ O) -O-, -O-C (═ O) -NH-, or substituted or unsubstituted C_1-6Alkylene or C_2-6Alkenylene chains in which up to two non-adjacent methylene units are independently optionally substituted by-C (═ O) -, -CO₂-、-C(＝O)C(＝O)-、-C(＝O)NR^L3-、-OC(＝O)-、-OC(＝O)NR^L3-、-NR^L3NR^L4-、-NR^L3NR^L4C(＝O)-、-NR^L3C(＝O)-、-NR^L3CO₂-、-NR^L3C(＝O)NR^L4-、-S(＝O)-、-SO₂-、-NR^L3SO₂-、-SO₂NR^L3-、-NR^L3SO₂NR^L4-, -O-, -S-, or-NR^L3-substitution; wherein R is^L3And R^L4Independently at each occurrence is hydrogen, alkyl, heteroalkyl, aryl, heteroaryl or acyl;

lxv) L is absent, -C (═ O), or substituted or unsubstituted C_1-6Alkylene or C_2-6Alkenylene chains in which up to two non-adjacent methylene units are independently optionally substituted by-C (═ O) -, -CO₂-、-C(＝O)C(＝O)-、-C(＝O)NR^L3-、-OC(＝O)-、-OC(＝O)NR^L3-、-NR^L3NR^L4-、-NR^L3NR^L4C(＝O)-、-NR^L3C(＝O)-、-NR^L3CO₂-、-NR^L3C(＝O)NR^L4-、-S(＝O)-、-SO₂-、-NR^L3SO₂-、-SO₂NR^L3-、-NR^L3SO₂NR^L4-, -O-, -S-, or-NR^L3-substitution; each occurrence of R therein^L3And R^L4Independently hydrogen, alkyl, heteroalkyl, aryl, heteroaryl, or acyl;

lxvi) L is absent;

lxvii) L is-C (═ O);

lxviii) L is absent, -C (═ O), -CH₂C(＝O)NH-、-CH₂NH-C(＝O)-、-O-CH₂-C(＝O)-、-CH₂-CH₂-C(＝O)-、-CH＝CH-C(＝O)NH-CH₂-、-CH(OH)-CH₂-O-、-CH(OH)-CH₂-N(CH₃)-、-CH(OH)-CH₂-CH₂-、-CH₂-CH₂-CH(OH)-、-O-CH₂-CH(OH)-、-O-CH₂-CH(OH)-CH₂-、-O-CH₂-CH₂-CH(OH)-、O-CH₂-CH₂-O-、-CH₂-CH₂-CH₂-O-、-CH₂-CH(OH)-CH₂-O、-CH₂-CH₂-O-、-CH-(CH₃)-NH-C(＝O)-、-CH₂-NH-SO₂-、-NH-SO₂-CH₂-、-CH₂-SO₂-NH-、-SO₂NH-CH₂-、-C(＝O)-NH-C(＝O)-、-NH-C(＝O)-NH-、-NH-C(＝O)-NH-CH₂-、-CH₂-NH-C(＝O)-NH-、-C(＝O)-NH-CH₂-C (═ O) -NH, -NH-C (═ O) -O-, or-O-C (═ O) -NH-;

lx) L is- (CH)₂)_q-, wherein q is 1 to 5;

lxx) L is-CH₂-；

Lxxi) L is- (CH₂)₃-；

Ixxii) L is a moiety having the structure

lxxii)AR¹Is one of the following structures:

wherein r is an integer from 0 to 6 at each occurrence; x₁、X₂、X₃And X₄Each independently is N or CR^Q1；AR₃Is a heterocyclic, aryl or heteroaryl moiety; r^Q1Independently at each occurrence is hydrogen, OR^Q3、OCF₃、SR^Q3Halogen, CN, isocyanate, NO₂、CF₃、NR^Q3QR^Q4、-SO₂R^Q3alkyl-NR^Q3R^Q4alkyl-C (═ O) -NR^Q3R^Q4alkyl-C (═ O) R^Q3Or an aliphatic, alicyclic, heteroaliphatic, heterocyclic, aryl, heteroaryl, alkylaryl, alkylheteroaryl, heteroalkylaryl, or heteroalkylheteroaryl moiety, wherein R is^Q3And R^Q4Independently at each occurrence is hydrogen, a protecting group, or an aliphatic, heteroaliphatic, aryl, or heteroaryl moiety; r^Q2Is hydrogen, an aliphatic, alicyclic, heteroaliphatic, heterocyclic, aryl, heteroaryl, alkylaryl, alkylheteroaryl, heteroalkylaryl, or heteroalkylheteroaryl moiety or a nitrogen protecting group;

lxxiv)AR¹is one of the following structures:

wherein r is an integer from 0 to 6 at each occurrence; x₁、X₂、X₃And X₄Independently is N or CR^Q1；X₅Is O, S or NR^Q2；AR³Is a heterocyclic, aryl or heteroaryl moiety; r^Q1Independently at each occurrence is hydrogen, OR^Q3、OCF₃、SR^Q3Halogen, CN, isocyanate, NO₂、CF₃、NR^Q3QR^Q4、-SO₂R^Q3alkyl-NR^Q3R^Q4alkyl-C (═ O) -NR^Q3R^Q4alkyl-C (═ O) R^Q3Or an aliphatic, alicyclic, heteroaliphatic, heterocyclic, aryl, heteroaryl, alkylaryl, alkylheteroaryl, heteroalkylaryl, or heteroalkylheteroaryl moiety, wherein R is^Q3And R^Q4Independently at each occurrence is hydrogen, a protecting group, or an aliphatic, heteroaliphatic, aryl, or heteroaryl moiety; and R^Q2Is hydrogen, aliphatic, alicyclic, heteroaliphatic, heterocyclic, aryl, heteroaryl, alkylaryl, alkylheteroaryl, heteroalkylAn aryl, or heteroalkyl heteroaryl moiety or a nitrogen protecting group;

lxxv)AR¹is one of the following structures:

wherein r is as defined above; x₁、X₂、X₃And X₄Independently is N or CH; x₅Is CHR^Q1Or NH; r^Q1Independently at each occurrence is hydrogen, OR^Q3、OCF₃、SR^Q3Halogen, CN, isocyanate, NO₂、CF₃、NR^Q3QR^Q4、-SO₂R^Q3alkyl-NR^Q3R^Q4alkyl-C (═ O) -NR^Q3R^Q4alkyl-C (═ O) R^Q3Or an aliphatic, alicyclic, heteroaliphatic, heterocyclic, aryl, heteroaryl, alkylaryl, alkylheteroaryl, heteroalkylaryl, or heteroalkylheteroaryl moiety, wherein R is^Q3And R^Q4Independently at each occurrence is hydrogen, a protecting group, or an aliphatic, heteroaliphatic, aryl, or heteroaryl moiety; and R^Q2Is hydrogen, an aliphatic, alicyclic, heteroaliphatic, heterocyclic, aryl, heteroaryl, alkylaryl, alkylheteroaryl, heteroalkylaryl, or heteroalkylheteroaryl moiety or a nitrogen protecting group;

lxxvi)AR¹is one of the following structures:

wherein X⁰Is F or Cl; x₂Is N or CR^Q1；X₅Is CH, O, S or NH; r^Q1Is hydrogen, methyl, -CF₃、-OCH₃、-COF₃Or halogen;

lxxvii)AR¹is as followsOne of the structures is as follows:

lxxviii)AR¹is one of the following structures:

lxxix)AR¹-L-is one of the following structures:

lxxx)AR¹-L-is one of the following structures:

lxxxi)R⁴independently at each occurrence is hydrogen, halogen, -CN, -NO₂Alkyl, alkylene, alkynyl, cycloalkyl, heteroalkyl, heterocyclic, aryl, heteroaryl, alkylaryl or alkylheteroaryl moieties, or is-GR^G1Wherein G is-O-, -S-, -NR^G2-、-CO-、-SO-、-SO₂-、-C(＝O)O-、-C(＝O)NR^G2-、-OC(＝O)-、-NR^G2C (═ O) -or-SO)₂NR^G2-，R^G1And R^G2Independently is a hydrogen, alkyl, alkylene, alkynyl, cycloalkyl, heteroalkyl, heterocyclic, aryl, heteroaryl, alkylaryl, or alkylheteroaryl moiety;

lxxxii)R⁴independently at each occurrence, hydrogen, halogen, or lower alkyl;

lxxxiii)R⁴independently at each occurrence is hydrogen or chloro;

lxxxiv) n is 0;

lxxxv) n is 2;

lxxxvi) n is 2 and R⁴At each occurrence is halogen;

lxxxvii) n is 2 and R⁴At each occurrence is Cl;

lxxxviii) p is 1;

lxxxix) p is 2; and/or

xc) Compounds of formula (II) wherein at-C (═ O) NHC (R)¹)(R²)R³When having the following structure:

AR¹is not one of

Wherein Y is¹Is N or CR^Q1；X¹、X²、X³And X⁴Independently is CR^Q1(ii) a X5 is NR^Q1O or S; r is 0 to 3; r^Q1Independently at each occurrence CN, NO₂Halogen, CF₃Alkyl, cycloalkyl, heteroalkyl, heterocyclic, aryl, heteroaryl, alkylaryl or alkylheteroaryl moieties or are GR^G1Wherein G is-O-, -S-, -NR^G2-、-CO-、-SO-、-C_0-6Alkyl SO₂-、-C_0-6Alkyl SO₂NR^G2-、-C(＝O)O-、-C(＝O)NR^G2-, -OC (═ O) -, or-NR^G2C(＝O)-，R^G1And R^G2Independently is a hydrogen, alkyl, cycloalkyl, heteroalkyl, heterocyclic, aryl, heteroaryl, alkylaryl or alkylheteroaryl moiety.

It will be understood that for each of the classes and subclasses described above and herein, the aliphatic or heteroaliphatic moiety, independently at any one or more occurrence, may be substituted or unsubstituted, cyclic or acyclic, straight or branched, and the aryl, heteroaryl, cycloaliphatic, cycloheteroaliphatic moiety, may be substituted or unsubstituted at any one or more occurrence.

The reader will also understand that the variables mentioned above under i) -to xc) (such as in particular R)¹、R²、R³、R⁴L, and AR¹) All possible combinations of (a) and (b) are considered to be part of the present invention. Thus, the invention includes passing the variable R¹、R²、R³、R⁴L, and AR¹Etc. and R for the above-mentioned i) -to xc)¹、R²、R³、R⁴、L、AR¹Etc. other variables/substituents as further defined (e.g., X)¹、X²、X³、X⁴、R^1A、R^2A、R^2C、R^2DEtc.) in any possible permutation to produce any and all compounds of formula I or II.

For example, exemplary combinations of the variables recited in I) through xc) above include those compounds of formula I wherein the following conditions are satisfied:

R¹and R²Each independently hydrogen, an amino acid side chain, - (CH)₂)_mOH、-(CH₂)_mAryl, - (CH)₂)_mHeteroaryl (wherein m is 0-6), -CH (R)^1A)(OR^1B)、-CH(R^1A)(NHR^1B) U-T-Q, or an aliphatic, alicyclic, heteroaliphatic or heteroalicyclic moiety optionally substituted with U-T-Q, wherein U is absent, -O-, -S (O)_0-2-、-SO₂N(R^1A)、-N(R^1A)-、-N(R^1A)C(＝O)-、-N(R^1A)C(＝O)-O-、-N(R^1A)C(＝O)-N(R^1B)-、-N(R^1A)-SO₂-, -C (═ O) -O-, -O-C (═ O) -, aryl, heteroaryl, alkylarylAlkyl heteroaryl, -C (═ O) -N (R)^1A)-、-O-C(＝O)-N(R^1A)-、-C(＝N-R^1E)-、-C(＝N-R^1E)-O-、-C(＝N-R^1E)-N(R^1A)-、-O-C(＝N-R^1E)-N(R^1A)-、-N(R^1A)C(＝N-R^1E)-、-N(R^1A)C(＝N-R^1E)-O-、N(R^1A)C(＝N-R^1E)-N(R^1B)-、-P(＝O)(OR^1A) -O-, or-P (═ O) (R)^1A) -O-; t is an absent, aliphatic, heteroaliphatic, aryl, heteroaryl, alkylaryl or alkylheteroaryl moiety; q is hydrogen, halogen, cyano, isocyanate, -OR^1B、-SR^1B；-N(R^1B)₂、-NHC(＝O)OR^1B、-NHC(＝O)N(R^1B)₂、-NHC(＝O)R^1B、-NHSO₂R^1B、-NHSO₂N(R^1B)₂、-NHSO₂NHC(＝O)OR^1B、-NHC(＝O)NHSO₂R^1B、-C(＝O)NHC(＝O)OR^1B、-C(＝O)NHC(＝O)R^1B、-C(＝O)NHC(＝O)N(R^1B)₂、-C(＝O)NHSO₂R^1B、-C(＝O)NHSO₂N(R^1B)₂、-C(＝S)N(R^1B)₂、-SO₂R^1B、-SO₂-O-R^1B、-SO₂-N(R^1B)₂、-SO₂-NHC(＝O)OR^1B、-SO₂-NHC(＝O)-N(R^1B)₂、-SO₂-NHC(＝O)R^1B、-O-C(＝O)N(R^1B)₂、-O-C(＝O)R^1B、-O-C(＝O)NHC(＝O)R^1B、-O-C(＝O)NH-SO₂R^1B、-O-SO₂R^1BOr an aliphatic, heteroaliphatic, aryl or heteroaryl moiety, or wherein R is¹And R²Together being a cycloaliphatic or heterocyclic moiety, or together beingWherein R is^1AAnd R^1BIndependently at each occurrence, hydrogen, aliphatic, alicyclic, heteroaliphaticHeterocyclic, aryl, heteroaryl, alkylaryl or alkylheteroaryl moiety, -COR^1Cor-CONR^1CR^1D(ii) a Wherein R is^1CAnd R^1DIndependently at each occurrence is hydrogen, hydroxy, or an aliphatic, heteroaliphatic, aryl, heteroaryl, alkylaryl, or alkylheteroaryl moiety; and R^1EIs a hydrogen, aliphatic, alicyclic, heteroaliphatic, heterocyclic, aryl, heteroaryl, alkylaryl OR alkylheteroaryl moiety, -CN, -OR^1C、-NR^1CR^1Dor-SO₂R^1C；

R³is-C (═ O) OR^3A、-C(＝O)H、-CH₂OR^3A、-CH₂O-C (═ O) -alkyl, -C (═ O) NH (R)^3A)、-CH₂X⁰(ii) a Wherein R is^3AIndependently at each occurrence is hydrogen, a protecting group, an aliphatic, alicyclic, heteroaliphatic, heteroalicyclic, aryl, heteroaryl, alkylaryl, alkylheteroaryl, heteroalkylaryl, or heteroalkylheteroaryl moiety, or R^3AAnd R¹Or R²Together form a heterocyclic moiety; wherein X⁰Is halogen selected from F, Cl, Br or I;

R⁴independently at each occurrence is hydrogen, halogen, -CN, -NO₂An aliphatic, alicyclic, heteroaliphatic, heteroalicyclic, aryl, heteroaryl, alkylaryl or alkylheteroaryl moiety, or is-GR^G1Wherein G is-O-, -S-, -NR^G2-、-CO-、-SO-、-SO₂-、-C(＝O)O-、-C(＝O)NR^G2-、-OC(＝O)-、-NR^G2C (═ O) -or-SO)₂NR^G2-，R^G1And R^G2Independently is a hydrogen, aliphatic, alicyclic, heteroaliphatic, heteroalicyclic, aryl, heteroaryl, alkylaryl, or alkylheteroaryl moiety;

AR¹aryl, heteroaryl, alkylaryl, alkylheteroaryl, alicyclic or heterocyclic moieties which are monocyclic or polycyclic;

A、B、d and E are connected by a single or double bond, where valency permits; wherein A, B, D and E are independently at each occurrence C-O, CRⁱRⁱⁱ、NRⁱ、CRⁱN, O, S, S (═ O) or SO₂(ii) a Wherein R isⁱIndependently at each occurrence is hydrogen, halogen, -CN, -NO₂An aliphatic, alicyclic, heteroaliphatic, heteroalicyclic, aryl, heteroaryl, alkylaryl or alkylheteroaryl moiety, or is-GR^G1Wherein G is-O-, -S-, -NR^G2-、-CO-、-SO-、-SO₂-、-C(＝O)O-、-C(＝O)NR^G2-、-OC(＝O)-、-NR^G2C (═ O) -or-SO)₂NR^G2-，R^G1And R^G2Independently is hydrogen, an aliphatic, alicyclic, heteroaliphatic, heteroalicyclic, aryl, heteroaryl, alkylaryl or alkylheteroaryl moiety, or any two adjacent RⁱTogether represent an alicyclic, heteroalicyclic, aryl, or heteroaryl moiety; and

l is absent or is V-W-X-Y-Z, wherein V, W, X, Y and Z are independently absent at each occurrence, C-O, NR^L1、-O-、-C(R^L1)＝、＝C(R^L1)-、-C(R^L1)(R^L2)、C(＝N-O-R^L1)、C(＝N-R^L1)、-N＝、S(O)_0-2(ii) a Substituted or unsubstituted C_1-6Alkylene or C_2-6Alkenylene chains in which up to two non-adjacent methylene units are independently optionally substituted by-C (═ O) -, -CO₂-、-C(＝O)C(＝O)-、-C(＝O)NR^L3-、-OC(＝O)-、-OC(＝O)NR^L3-、-NR^L3NR^L4-、-NR^L3NR^L4C(＝O)-、-NR^L3C(＝O)-、-NR^L3CO₂-、-NR^L3C(＝O)NR^L4-、-S(＝O)-、-SO₂-、-NR^L3SO₂-、-SO₂NR^L3-、-NR^L3SO₂NR^L4-, -O-, -S-, or-NR^L3-substitution; wherein R is^L3And R^L4Independently at each occurrence is hydrogen, alkyl, heteroalkyl, aryl, heteroalkylAryl or acyl; or is an aliphatic, alicyclic, heteroaliphatic, heteroalicyclic, aryl, heteroaryl, alkylaryl or alkylheteroaryl moiety; r^L1And R^L2Independently at each occurrence is hydrogen, hydroxy, protected hydroxy, amino, protected amino, thio, protected thio, halogen, cyano, isocyanate, carboxy, carboxyalkyl, formyl, formyloxy, azido, nitro, ureido, thioureido, thiocyanato, alkoxy, aryloxy, mercapto, sulfonamido, benzamido, tosyl, or an aliphatic, cycloaliphatic, heteroaliphatic, aryl, heteroaryl, alkylaryl, or alkylheteroaryl moiety, or wherein R is^L1And R^L2In one or more occurrences, or together with V, W, X, Y or one of Z, form an alicyclic or heterocyclic moiety or form an aryl or heteroaryl moiety.

Other exemplary combinations are illustrated by the following subclasses I through XIV of compounds:

I) a compound (and pharmaceutically acceptable derivatives thereof) having the structure:

wherein R is^4AAnd R^4BIndependently is a halogen selected from F, Cl, Br or I; and R^B1、R^B2And R^EIndependently hydrogen or substituted or unsubstituted lower alkyl. In certain embodiments, R^4AAnd R^4BEach is Cl. In other embodiments, R^B1And R^B2One is hydrogen and the other is substituted or unsubstituted lower alkyl. In certain exemplary embodiments, R^B1And R^B2Each is hydrogen. In certain other exemplary embodiments, R^B1And R^B2Each is a lower alkyl group. In certain exemplary embodiments, R^B1And R^B2Each is methyl. In other embodiments, R^EIs hydrogen. In thatIn other embodiments, R^EIs substituted or unsubstituted lower alkyl. In other embodiments, R^EIs substituted or unsubstituted methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, n-pentyl, sec-pentyl, tert-pentyl or n-hexyl. In certain embodiments, R^4AAnd R^4BEach is Cl; and R^B1And R^B2Each is hydrogen.

II) a compound (and pharmaceutically acceptable derivatives thereof) having the structure:

wherein R is^4AAnd R^4BIndependently is a halogen selected from F, Cl, Br or I; and R^EIs hydrogen or substituted or unsubstituted lower alkyl. In certain embodiments, R^4AAnd R^4BEach is Cl. In other embodiments, R^EIs hydrogen. In other embodiments, R^EIs substituted or unsubstituted lower alkyl. In other embodiments, R^EIs substituted or unsubstituted methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, n-pentyl, sec-pentyl, tert-pentyl or n-hexyl.

III) Compounds having the following structure (and pharmaceutically acceptable derivatives thereof):

wherein R is^4AAnd R^4BIndependently is a halogen selected from F, Cl, Br or I; and R^EIs hydrogen or substituted or unsubstituted lower alkyl. In certain embodiments, R^4AAnd R^4BEach is Cl. In other embodiments, R^EIs hydrogen. In other embodiments, R^EIs substituted or unsubstituted lower alkyl. In other embodiments, R^EIs substituted or unsubstituted methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, n-pentyl, sec-pentyl, tert-pentyl or n-hexyl.

IV) Compounds having the following structure (and pharmaceutically acceptable derivatives thereof):

wherein R is^4AAnd R^4BIndependently is a halogen selected from F, Cl, Br or I; and R^EIs hydrogen or substituted or unsubstituted lower alkyl. In certain embodiments, R^4AAnd R^4BEach is Cl. In other embodiments, R^EIs hydrogen. In other embodiments, R^EIs substituted or unsubstituted lower alkyl. In other embodiments, R^EIs substituted or unsubstituted methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, n-pentyl, sec-pentyl, tert-pentyl or n-hexyl.

V) a compound (and pharmaceutically acceptable derivatives thereof) having the structure:

wherein R is^4AAnd R^4BIndependently is a halogen selected from F, Cl, Br or I; r^AIs hydrogen, lower alkyl or acyl; and R^EIs hydrogen or substituted or unsubstituted lower alkyl. In certain embodiments, R^4AAnd R^4BEach is Cl. In other embodiments, R^EIs hydrogen. In other embodiments, R^EIs substituted or unsubstituted lower alkyl. In other embodiments, R^EIs substituted or unsubstituted methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, n-pentylSec-amyl, tert-amyl or n-hexyl.

VI) a compound (and pharmaceutically acceptable derivatives thereof) having the structure:

wherein R is^4AAnd R^4BIndependently is a halogen selected from F, Cl, Br or I; r^A1、R^A2、R^B1And R^B2Independently hydrogen or substituted or unsubstituted lower alkyl. In certain embodiments, R^4AAnd R^4BEach is Cl. In certain embodiments, R^A1、R^A2、R^B1And R^B2Each is hydrogen.

VII) compounds (and pharmaceutically acceptable derivatives thereof) having the structure:

wherein R is^4AAnd R^4BIndependently is a halogen selected from F, Cl, Br or I; and R^AAnd R^BIndependently hydrogen or substituted or unsubstituted lower alkyl. In certain embodiments, R^4AAnd R^4BEach is Cl. In certain embodiments, R^AAnd R^BEach is hydrogen.

VIII) compounds having the structure (and pharmaceutically acceptable derivatives thereof):

wherein R is^4AAnd R^4BIndependently is a halogen selected from F, Cl, Br or I; and R^AIs hydrogen or substituted or unsubstituted lower alkyl. In certain embodiments, R^4AAnd R^4BEach is Cl. In certain embodiments, R^AIs hydrogen.

IX) compounds having the structure (and pharmaceutically acceptable derivatives thereof):

wherein R is^4AAnd R^4BIndependently is a halogen selected from F, Cl, Br or I; and R^BIs hydrogen or substituted or unsubstituted lower alkyl. In certain embodiments, R^4AAnd R^4BEach is Cl. In certain embodiments, R^BIs hydrogen.

X) a compound (and pharmaceutically acceptable derivatives thereof) having the structure:

wherein R is^4AAnd R^4BIndependently is a halogen selected from F, Cl, Br or I; and R^AIs hydrogen or substituted or unsubstituted lower alkyl. In certain embodiments, R^4AAnd R^4BEach is Cl. In certain embodiments, R^AIs hydrogen.

XI) compounds having the structure (and pharmaceutically acceptable derivatives thereof):

wherein R is^4AAnd R^4BIndependently is a halogen selected from F, Cl, Br or I; r^A、R^BAnd R^EIndependently hydrogen or substituted or unsubstituted lower alkyl. In certain embodiments, R^4AAnd R^4BEach is Cl. In certain embodiments, R^AAnd R^BEach is hydrogen. At a certain pointIn some other embodiments, R^EIs hydrogen. In other embodiments, R^A、R^BAnd R^EEach is hydrogen.

XII) a compound having the structure (and pharmaceutically acceptable derivatives thereof):

wherein R is^4AAnd R^4BIndependently is a halogen selected from F, Cl, Br or I; r^A、R^BAnd R^EIndependently hydrogen or substituted or unsubstituted lower alkyl. In certain embodiments, R^4AAnd R^4BEach is Cl. In certain embodiments, R^AAnd R^BEach is hydrogen. In certain other embodiments, R^EIs hydrogen. In other embodiments, R^A、R^BAnd R^EEach is hydrogen.

XIII) a compound (and pharmaceutically acceptable derivatives thereof) having the structure:

wherein R is^4AAnd R^4BIndependently is a halogen selected from F, Cl, Br or I; and A and B are independently N or CH. In certain embodiments, R^4AAnd R^4BEach is Cl. In certain embodiments, a is N. In certain embodiments, a is CH. In certain embodiments, B is N. In certain embodiments, a is CH. In certain embodiments, a and B are each N. In certain embodiments, a is CH. In certain embodiments, a and B are each CH.

XIV) a compound (and pharmaceutically acceptable derivatives thereof) having the structure:

wherein R is^4AAnd R^4BIndependently is a halogen selected from F, Cl, Br or I; and A and B are independently N or CH. In certain embodiments, R^4AAnd R^4BEach is Cl. In certain embodiments, a is N. In certain embodiments, a is CH. In certain embodiments, B is N. In certain embodiments, a is CH. In certain embodiments, a and B are each N. In certain embodiments, a is CH. In certain embodiments, a and B are each CH.

In certain embodiments, for the compounds of classes I-XIV described above, AR¹-L-is a moiety having one of the following structures:

and-C (═ O) NHC (R)¹)(R²)R³Is a moiety having one of the following structures:

or a bioisostere thereof;

wherein R is^2AAnd R^3AAs defined in classes and subclasses herein; and R^2DIs a moiety having one of the following structures:

wherein s is an integer from 0 to 6; r^P1Independently at each occurrence is hydrogen, halogen, CN, isocyanate, NO₂、-P(＝O)(YR^P5)₂Alkyl, cycloalkyl, heteroalkyl, heterocyclic, aryl, heteroaryl, alkylaryl orAn alkylheteroaryl moiety, or is-GR^G1Wherein G is-O-, -S-, -NR^G2-、-CO-、-SO-、-SO₂-、-C(＝O)O-、-C(＝O)NR^G2-、-OC(＝O)-、-NR^G2C (═ O) -or-SO)₂NR^G2-，R^G1And R^G2Independently is a hydrogen, alkyl, cycloalkyl, heteroalkyl, heterocyclic, aryl, heteroaryl, alkylaryl or alkylheteroaryl moiety; y is independently at each occurrence a bond or O; r^P5Independently at each occurrence is alkyl, heteroalkyl, aryl or heteroaryl, or R when Y is O^P5May also be hydrogen; r^P2Independently at each occurrence is hydrogen, an aliphatic, alicyclic, heteroaliphatic, heterocyclic, aryl, heteroaryl, alkylaryl, alkylheteroaryl, heteroalkylaryl, or heteroalkylheteroaryl moiety or a nitrogen protecting group; wherein any two adjacent R^P1And R^P2Together may form a cycloalkyl, heterocyclic, aryl or heteroaryl moiety.

In certain embodiments, R^2AAnd R^3AEach is hydrogen.

In certain embodiments, R^2DIs a moiety having one of the following structures:

wherein R is^P1Independently at each occurrence is hydrogen, halogen, methyl, -OCH₃、-OH、-NH₂、-NHCH₃or-N (CH)₃)₂。

In certain embodiments, R^2DIs a moiety having one of the following structures:

it should also be understood that for each of the above subclasses I-XIV, a variety of other subclasses are of particular interest, including but not limited to those classes I) -xc) above and the classes, subclasses, and species described above and in the examples herein.

Some of the aforementioned compounds may include one or more asymmetric centers and, thus, may exist in multiple isomeric forms, e.g., stereoisomers and/or diastereomers. Thus, the compounds of the present invention and pharmaceutical compositions thereof may be in the form of individual enantiomers, diastereomers or geometric isomers, or may be in the form of mixtures of stereoisomers. In certain embodiments, the compounds of the invention are enantiomerically pure compounds. In certain other embodiments, mixtures of stereoisomers or diastereomers are provided.

In addition, certain compounds, as described herein, may have one or more double bonds, which may exist as either Z or E isomers unless otherwise specified. The invention additionally includes compounds that are individual isomers substantially free of other isomers or that are mixtures of different isomers, such as racemic mixtures of stereoisomers. In addition to the compounds themselves, the present invention also includes pharmaceutically acceptable derivatives of these compounds, compositions comprising one or more compounds of the present invention and one or more pharmaceutically acceptable excipients or additives.

The compounds of the invention may be prepared by crystallization of a compound of formula (I) or (II) under different conditions and may exist as one polymorph or a combination of polymorphs of a compound of formula (I) or (II) which form part of the present invention. For example, different solvents, or different mixtures of solvents, for recrystallization may be used; by crystallization at different temperatures; different polymorphs are identified and/or prepared by different cooling regimes from very fast to very slow cooling during crystallization. Polymorphs can also be obtained by heating or melting the compound followed by gradual or rapid cooling. The presence of polymorphs can be determined by solid state detection nuclear magnetic resonance spectroscopy, infrared spectroscopy, differential scanning calorimetry, powder X-ray diffraction patterns, and/or other techniques. Accordingly, the present invention includes the compounds of the present invention, their derivatives, their tautomeric forms, their stereoisomers, their polymorphs, their pharmaceutically acceptable salts, their pharmaceutically acceptable solvates, and pharmaceutically acceptable compositions containing them.

2) Pharmaceutical compositions

As noted above, the present invention provides novel compounds having biological properties useful in the treatment of Mac-1 and LFA-1 mediated diseases.

Accordingly, in another aspect of the invention there is provided a pharmaceutical composition comprising any one of the compounds described herein (or a prodrug, pharmaceutically acceptable salt or other pharmaceutically acceptable derivative thereof), and optionally a pharmaceutically acceptable carrier. In certain embodiments, the compositions optionally further comprise one or more additional therapeutic agents. Alternatively, a compound of the invention may be administered to a patient in need thereof in combination with the administration of one or more other therapeutic agents. For example, the additional therapeutic agent for Administration in combination with or contained within a pharmaceutical composition of a compound of the present invention may be an approved anti-inflammatory agent, or it may be any of a number of drugs that have undergone Food and Drug Administration approval and ultimately obtained approval for the treatment of any disease mediated by Mac-1 or LFA-1. It will also be appreciated that certain compounds of the invention may exist in the free state for use in therapy, or as pharmaceutically acceptable derivatives thereof where appropriate.

As noted above, the pharmaceutical compositions of the present invention additionally include a pharmaceutically acceptable carrier, as used herein, which includes any and all solvents, diluents, or other liquid media, dispersing or suspending aids, surfactants, isotonicity agents, thickening or emulsifying agents, preservatives, solid binders, lubricants and the like, as appropriate for the particular dosage form desired. Remington's pharmaceutical Sciences, Sixteenth Edition, e.w. martin (Mack Publishing co., Easton, Pa.,1980) discloses a variety of carriers and known manufacturing techniques for formulating pharmaceutical compositions. Unless any conventional carrier medium is incompatible with the compounds of the present invention, such as to produce any undesirable biological effect or to interact in a deleterious manner with any other component of the pharmaceutical composition, its use is contemplated to be within the scope of the present invention. Some examples of materials that can be used as pharmaceutically acceptable carriers include, but are not limited to, sugars such as lactose, glucose, and sucrose; starches, such as corn starch and potato starch; cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; packaging with powder of Astragalus membranaceus gum; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols, such as propylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffering agents such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; ringer's solution; ethanol, and phosphate buffer, as well as other non-toxic compatible lubricants such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, detackifying agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the composition, according to the judgment of the formulator.

Liquid dosage forms for oral administration include, but are not limited to, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active compounds, the liquid dosage forms may contain inert diluents commonly used in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1, 3-butylene glycol, dimethylformamide, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof. In addition to inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

Injectable preparations such as sterile injectable aqueous or oleaginous suspensions may be formulated according to the known art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution, suspension or emulsion in a parenterally-acceptable, non-toxic diluent or solvent, for example as a solution in 1, 3-butanediol. Acceptable media and solvents that can be employed are water, Ringer's solution, U.S. p. and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil may be employed including synthetic mono-or diglycerides. In addition, fatty acids such as oleic acid find use in the preparation of injectables.

Injectable formulations can be sterilized, for example, by filtration through a bacterial-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions that can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use.

In order to prolong the effect of a drug, it is often desirable to delay the absorption of the drug from subcutaneous or intramuscular injection. This can be achieved by using liquid suspensions or crystalline or amorphous materials with poor water solubility. The rate of absorption of the drug then depends on its rate of dissolution, which in turn may depend on crystal size and crystal form. Alternatively, delayed absorption of a parenterally administered pharmaceutical form may be achieved by dissolving or suspending the drug in an oily medium. Injectable depot forms are produced by forming a microcapsule matrix of the drug in a biodegradable polymer such as polylactide-polyglycolic acid. Depending on the ratio of drug to polymer and the nature of the particular polymer employed, the rate of drug release can be controlled. Examples of other biodegradable polymers include (poly (orthoesters) and poly (anhydrides) depot injectable formulations are also prepared by entrapping the drug in liposomes or microemulsions which are compatible with body tissues.

Compositions for rectal or vaginal administration are preferably suppositories which can be prepared by mixing the compounds of the invention with suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol or a suppository wax which are solid at ambient temperature but liquid at body temperature and therefore melt in the rectal or vaginal cavity and release the active compound.

Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound is admixed with at least one inert, pharmaceutically acceptable excipient or carrier such as sodium citrate or dicalcium phosphate and/or a) fillers or extenders such as starches, lactose, sucrose, glucose, mannitol, and silicic acid, b) binders such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and acacia, c) humectants such as glycerol, d) disintegrating agents such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, e) solution retarding agents such as paraffin, f) absorption promoters such as quaternary ammonium compounds, g) wetting agents such as, for example, hexadecanol and glyceryl monostearate, h) adsorbents such as kaolin and bentonite, and i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, including, Sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents.

Solid compositions of a similar type may also be employed as fillers in filled soft and hard gelatin capsules using excipients such as lactose and high molecular weight polyethylene glycols and the like. Solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared using coatings and shells such as enteric coatings and other coatings well known in the pharmaceutical formulating art. They may optionally contain opacifying agents and may also be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally in a delayed manner. Examples of embedding compositions that can be used include polymers and waxes. Solid compositions of a similar type may also be employed as fillers in filled soft and hard gelatin capsules using excipients such as lactose and high molecular weight polyethylene glycols and the like.

The active compound may also be in microencapsulated form with one or more excipients as described above. Solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared using coatings and shells such as enteric coatings, release controlling coatings, and other coatings well known in the pharmaceutical formulating art. In such solid dosage forms, the active compound may be mixed with at least one inert diluent such as sucrose, lactose and starch. Such dosage forms may also include additional substances other than inert diluents, e.g., tablet lubricants and other tablet aids such as magnesium stearate and microcrystalline cellulose, as is normal in manufacturing. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents. They may optionally contain opacifying agents and may also be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally in a delayed manner. Examples of embedding compositions that can be used include polymers and waxes.

The present invention includes pharmaceutically acceptable topical formulations of the compounds of the present invention. As used herein, the term "pharmaceutically acceptable topical formulation" refers to any formulation that is pharmaceutically acceptable for intradermal administration of the compounds of the invention by applying the formulation to the epidermis. In certain embodiments of the invention, the topical formulation includes a carrier system. Pharmaceutically effective carriers include, but are not limited to, solvents (e.g., alcohols, polyols, water), creams, lotions, ointments, oils, patches, liposomes, powders, emulsions, microemulsions, and buffered solutions (e.g., hypotonic or buffered saline) or any other carrier known in the art for topical administration of drugs. A more exhaustive list of carriers known in the art is provided by standard references in the art, such as Remington's pharmaceutical Sciences, 16 th edition, 1980 and 17 th edition, 1985, both published by Mack publishing company, Easton, Pa., the disclosure of which is incorporated herein by reference in its entirety. In certain other embodiments, the topical formulations of the present invention may include an excipient. The pharmaceutically acceptable topical formulations of the present invention can be prepared using any pharmaceutically acceptable excipient known in the art. Examples of excipients that may be included in the topical formulations of the present invention include, but are not limited to, preservatives, antioxidants, moisturizers, emollients, buffers, solubilizers, other penetrants, skin protectants, surfactants, and propellants and/or additional therapeutic agents for use in combination with the compounds of the present invention. Suitable preservatives include, but are not limited to, alcohols, quaternary amines, organic acids, parabens, and phenols. Suitable antioxidants include, but are not limited to, ascorbic acid and its esters, sodium bisulfite, butylated hydroxytoluene, butylated hydroxyanisole, vitamin E, and chelating agents such as EDTA and citric acid. Suitable moisturizers include, but are not limited to, glycerin, sorbitol, polyethylene glycol, urea, and propylene glycol. Suitable buffers for use in the present invention include, but are not limited to, buffers of citric acid, hydrochloric acid, and lactic acid. Suitable solubilizers include, but are not limited to, quaternary ammonium chlorides, cyclodextrins, benzyl benzoate, lecithin, and polysorbates. Suitable skin care agents that may be used in the topical formulations of the present invention include, but are not limited to, vitamin E oil, allantoin, dimethicone, glycerin, petrolatum, and zinc oxide.

In certain embodiments, a pharmaceutically acceptable topical formulation of the invention comprises at least one compound of the invention and a penetration enhancer. The choice of topical formulation depends on several factors, including the condition to be treated, the physicochemical properties of the compounds of the invention and other excipients, their stability in the formulation, available processing equipment, and cost constraints. As used herein, the term "penetration enhancer" refers to an agent capable of transporting a pharmacologically active compound through the stratum corneum and into the epidermis or dermis, preferably with little or no systemic absorption. A variety of compounds have been evaluated for their effectiveness in enhancing the rate of drug permeation through the skin. See, e.g., Percutaneous pennetration Enhancers, maibachh.i., and Smith h.e. (eds.), CRC Press, inc., Boca Raton, Fla (1995), which examined the use and testing of various skin permeation Enhancers; and Buyukttim et al, Chemical Means of Transdermal Drug evaluation in Transdermal and Topical Drug Delivery Systems, GoshT.K., Pfister W.R., Yum S.I, (Eds.), Interpharm Press Inc., Buffalo Grove, Ill. (1997). In certain exemplary embodiments, penetrants for use in the present invention include, but are not limited to, triglycerides (e.g., soybean oil), aloe vera compositions (e.g., aloe-vera gel), ethanol, isopropanol, octoly phenyl polyethylene glycol, oleic acid, polyethylene glycol 400, propylene glycol, N-dodecyl methyl sulfoxide, fatty acid esters (e.g., isopropyl myristate, methyl laurate, glycerol monooleate, and propylene glycol monooleate), and N-methyl pyrrolidone.

In certain embodiments, the composition may be in the form of an ointment, paste, cream, lotion, gel, powder, solution, spray, inhalant, or patch. In certain exemplary embodiments, the formulation of the composition of the present invention is a cream, which may additionally comprise saturated or unsaturated fatty acids such as stearic acid, palmitic acid, oleic acid, palmitoleic acid, cetyl alcohol or oleyl alcohol, with stearic acid being particularly preferred. The cream of the present invention may also contain a non-ionic surfactant, such as poly-40-hydroxy stearate. In certain embodiments, the active ingredient is mixed under sterile conditions with a pharmaceutically acceptable carrier and any required preservatives or buffers that may be required. Ophthalmic formulations, ear drops, and eye drops are also contemplated as being within the scope of the present invention. In addition, the present invention contemplates the use of transdermal patches, which have the additional advantage of providing controlled delivery of the compound to the body. Such dosage forms are produced by dissolving or dispensing the compound in a suitable medium. As noted above, permeation enhancers may also be used to increase the flux of a compound through the skin. The rate can be controlled by providing a rate controlling membrane or by dispersing the compound in a polymer matrix or gel.

It is also to be understood that the compounds and pharmaceutical compositions of the present invention can be formulated and used in combination therapy, that is, the compounds and pharmaceutical compositions can be formulated with, administered concurrently with, before, or after one or more other desired therapies or treatments. The particular combination therapy (therapy or procedure) used in the combination regimen takes into account the compatibility of the desired therapy and/or procedure and the therapeutic effect desired to be achieved. It is also understood that the treatments employed may achieve the desired effect for the same disease (e.g., the compounds of the invention may be administered simultaneously with another anti-inflammatory agent), or they may achieve different effects (e.g., control of any side effects).

In certain embodiments, the pharmaceutical compositions of the invention additionally comprise one or more additional therapeutically active components (e.g., anti-inflammatory and/or palliative agents). For the purposes of the present invention, the term "palliative" is meant to focus on alleviating the symptoms of the disease and/or the side effects of the treatment regimen and is not intended for therapeutic treatment. For example, palliative therapy includes analgesics, antiemetics, and anti-nausea drugs.

3) Research applications, pharmaceutical applications and methods of treatment

Research applications

In accordance with the present invention, the compounds of the present invention may be tested in any available assay known in the art for identifying compounds having the ability to modulate adhesion between an intracellular adhesion molecule and the leukocyte integrin family of receptors, the ability to antagonize the leukocyte-associated CD11/CD18 receptor, and/or the ability to antagonize Mac-1 and/or LFA-1. For example, the assay may be cellular or acellular, in vivo or in vitro, in high-throughput or low-throughput format, and the like.

Thus, in one aspect, compounds of the invention of particular interest include those in which the following conditions are satisfied:

modulating adhesion between intracellular adhesion molecules (e.g., ICAM-1, -2, and-3) and the leukocyte integrin family of receptors;

exhibit the ability to antagonize the leukocyte-associated CD11/CD18 receptor;

exhibit the ability to antagonize Mac-1 and/or LFA-1; and

useful in the treatment of LFA-1 mediated diseases.

As detailed in the examples herein, certain compounds of the invention exhibit IC in assays that determine the ability of compounds to modulate T cell adhesion to 5dICAM-Ig (e.g., cell attachment assays)₅₀The value is less than or equal to 50 mu M. In thatIn certain other embodiments, the compounds of the present invention exhibit IC₅₀The value is less than or equal to 40 mu M. In certain other embodiments, the compounds of the present invention exhibit IC₅₀The value is less than or equal to 30 mu M. In certain other embodiments, the compounds of the present invention exhibit IC₅₀The value is less than or equal to 20 mu M. In certain other embodiments, the compounds of the present invention exhibit IC₅₀The value is less than or equal to 10 mu M. In certain other embodiments, the compounds of the present invention exhibit IC₅₀The value is less than or equal to 7.5 mu M. In certain embodiments, the compounds of the present invention exhibit IC₅₀The value is less than or equal to 5 mu M. In certain other embodiments, the compounds of the present invention exhibit IC₅₀The value is less than or equal to 2.5 mu M. In certain embodiments, the compounds of the present invention exhibit IC₅₀The value is less than or equal to 1 mu M. In certain other embodiments, the compounds of the present invention exhibit IC₅₀The value is less than or equal to 750 nM. In certain other embodiments, the compounds of the present invention exhibit IC₅₀The value is less than or equal to 500 nM. In certain other embodiments, the compounds of the present invention exhibit IC₅₀The value is less than or equal to 250 nM. In certain other embodiments, the compounds of the present invention exhibit IC₅₀The value is less than or equal to 100 nM. In other embodiments, exemplary compounds exhibit IC₅₀The value is less than or equal to 75 nM. In other embodiments, exemplary compounds exhibit IC₅₀The value is less than or equal to 50 nM. In other embodiments, exemplary compounds exhibit IC₅₀The value is less than or equal to 40 nM. In other embodiments, exemplary compounds exhibit IC₅₀The value is less than or equal to 30 nM. In other embodiments, exemplary compounds exhibit IC₅₀The value is less than or equal to 20 nM. In other embodiments, exemplary compounds exhibit IC₅₀The value is less than or equal to 10 nM. In other embodiments, exemplary compounds exhibit IC₅₀The value is less than or equal to 5 nM.

Pharmaceutical use and method of treatment

As noted above, certain compounds described herein exhibit activity as modulators of adhesion between intracellular adhesion molecules in general. More specifically, the compounds of the present invention exhibit the ability to antagonize CD11/CD18 receptors associated with leukocytes, and in certain embodiments, exhibit the ability to antagonize LFA-1 interactions. Thus, in certain embodiments, the compounds of the invention are useful for treating LFA-1 mediated diseases.

Thus, in another aspect of the invention, there is provided a method for the treatment (or prevention) of LFA-1 mediated diseases, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of formula (I) or (II) as described herein. In certain embodiments, methods are provided for treating LFA-1 mediated diseases comprising administering to a subject in need thereof a therapeutically effective amount of a compound of the present invention or a pharmaceutical composition comprising a compound of the present invention in an amount and for a time necessary to achieve the desired effect.

In certain embodiments, the method involves administering to a subject (including but not limited to a human or animal) in need thereof a therapeutically effective amount of a compound or a pharmaceutically acceptable derivative thereof.

As noted above, the present invention provides novel compounds having biological properties useful for the treatment of Mac-1 and/or LFA-1 mediated diseases. In certain embodiments, the compounds of the invention are useful for treating psoriasis, responses associated with inflammatory bowel disease (e.g., crohn's disease and ulcerative colitis), dermatitis, meningitis, encephalitis, uveitis, allergic conditions such as eczema and asthma, conditions involving T-cell infiltration and chronic inflammatory responses, skin hypersensitivity reactions (including poison ivy and poison oak), atherosclerosis, autoimmune diseases such as rheumatoid arthritis, Systemic Lupus Erythematosus (SLE), diabetes, multiple sclerosis, Reynaud's syndrome, autoimmune thyroiditis, experimental autoimmune encephalomyelitis, sjogren's syndrome, juvenile onset diabetes, delayed-onset allergic-related immune responses mediated by cytokines and T lymphocytes typically found in tuberculosis, sarcoidosis, polymyositis, granulomatosis and vasculitis, Pernicious anemia, diseases involving leukocyte extravasation, CNS inflammatory diseases, sepsis or multiple organ damage syndrome secondary to trauma, autoimmune hemolytic anemia, myasthenia gravis, antigen-antibody complex mediated diseases, all types of transplantation, including graft versus host disease or host versus graft disease, HIV and rhinovirus infections, and pulmonary fibrosis, to name a few.

As described in more detail herein, in general, the compounds of the invention are useful as antagonists of the interaction between intracellular adhesion molecules (e.g., ICAM-1, -2, or-3) and the leukocyte integrin family of receptors. Thus, in certain embodiments, the present invention provides compounds useful for treating diseases mediated by the CD11/CD18 family of cell adhesion molecules. In certain embodiments of particular interest, the present invention provides compounds useful for treating diseases mediated by Mac-1 and/or LFA-1. For example, the compounds of the present invention may be particularly useful in the treatment of inflammatory diseases, organ transplant rejection, and autoimmune diseases, to name a few.

Thus, as mentioned above, in another aspect of the invention there is provided a method for the treatment of a disease mediated by the CD11/CD18 family of cell adhesion molecules, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of formula (I) or (II). In certain embodiments of particular interest, the methods of the invention are useful for treating diseases mediated by Mac-1 or LFA-1. It is understood that the compounds and compositions can be administered in any amount and by any route of administration effective to treat diseases mediated by the CD11/CD18 family of cell adhesion molecules in accordance with the methods of the present invention. For example, in certain exemplary embodiments, the compounds of the invention are useful as antagonists of the interaction between Mac-1 or LFA-1 and intracellular adhesion molecules (e.g., ICAM-1), and thus, the compounds are useful in the treatment of LFA-1 mediated diseases, including, but not limited to, psoriasis, responses associated with inflammatory bowel disease (e.g., crohn's disease and ulcerative colitis), dermatitis, meningitis, encephalitis, uveitis, allergic conditions such as eczema and asthma, conditions involving T cell infiltration and chronic inflammatory responses, skin hypersensitivity reactions (including poison ivy and poison oak), atherosclerosis, autoimmune diseases such as rheumatoid arthritis, Systemic Lupus Erythematosus (SLE), diabetes, multiple sclerosis, Reynaud's syndrome, autoimmune thyroiditis, experimental autoimmune encephalomyelitis, Sjogren's syndrome, juvenile onset diabetes, and immune responses typically found in tuberculosis associated with delayed hypersensitivity mediated by cytokines and T-lymphocytes, sarcoidosis, polymyositis, granulomatosis and vasculitis, pernicious anemia, diseases involving leukocyte extravasation, CNS inflammatory diseases, multiple organ damage syndrome secondary to sepsis or trauma, autoimmune hemolytic anemia, myasthenia gravis, antigen-antibody complex mediated diseases, all types of transplantation including graft versus host disease or host versus graft disease, HIV and rhinovirus infections, pulmonary fibrosis, to name a few. Thus, as used herein, the expression "effective amount" refers to an amount of a drug sufficient to antagonize the interaction between an intracellular adhesion molecule (e.g., ICAM) and the leukocyte integrin family of receptors and to exhibit a therapeutic effect. The exact amount required will vary from subject to subject, depending on the species, age, and general condition of the subject, the severity of the infection, the particular therapeutic agent, mode of administration, and the like. The compounds of the present invention are preferably formulated in dosage unit form for ease of administration and uniformity of dosage. As used herein, the expression "dosage unit form" refers to a physically discrete unit of a therapeutic agent suitable for the patient to be treated. It will be understood, however, that the total daily amount of the compounds and compositions of the present invention will be determined by the attending physician, within the scope of sound medical judgment. The specific therapeutically effective dose level for any particular patient or organism will depend upon a variety of factors including the disease to be treated and the severity of the disease; the activity of the particular compound employed; the specific composition employed; the age, weight, general health, sex, and diet of the patient; the time of administration, route of administration, and rate of excretion of the particular compound employed; the duration of the treatment; medicaments for administration in combination or concomitantly with the specific compound employed; various factors are widely known in The medical field (see, e.g., Goodman and Gilman's, "The pharmaceutical Basis of Therapeutics", tenth edition, A.Gilman, J.Hardman and L.Limbird, eds., McGraw-Hill Press, 155-.

Another aspect of the invention relates to a method of inhibiting the interaction between LFA-1 and ICAM-1 in a biological sample or a patient, comprising administering to the patient, or contacting said biological sample with a compound of formula I or II or a composition comprising said compound.

Another aspect of the invention relates to a method of inhibiting the interaction of CD11a and/or CD18 with ICAM-1, ICAM-2, or ICAM-3 in a biological sample or a patient, comprising administering to the patient, or contacting said biological sample with a compound of formula I or II, or a composition comprising said compound.

In addition, the pharmaceutical compositions of the present invention may be administered to humans and other animals orally, rectally, parenterally, intracisternally (intracisternally), intravaginally, intraperitoneally, topically (as by powders, ointments, or drops), buccally (as an oral or nasal spray), after formulation in the required dosage form with an appropriate pharmaceutically acceptable carrier, depending on the severity of the infection to be treated. In certain embodiments, a compound of the invention may be administered once or more times daily at a dosage level of from about 0.001mg/kg to about 50mg/kg body weight of the subject per day, from about 0.01mg/kg to about 25mg/kg body weight of the subject per day, or from about 0.1mg/kg to about 10mg/kg body weight of the subject per day to achieve the desired therapeutic effect. It is also understood that a dose of less than 0.001mg/kg or greater than 50mg/kg (e.g., 50-100mg/kg) may be administered to a subject. In certain embodiments, the compound is administered orally or parenterally.

Therapeutic medicine bag

In other embodiments, the invention relates to kits for conveniently and efficiently carrying out the methods of the invention. Generally, a pharmaceutical pack comprises one or more containers filled with one or more of the components of the pharmaceutical composition of the invention. Such kits are particularly suitable for delivering solid oral dosage forms such as tablets or capsules. Preferably such kits comprise a plurality of unit doses and may further comprise a card having the doses oriented in their intended order of application. If desired, memory aids, such as numbers, letters, or other indicia forms, or the use of calendar inserts, can be provided to specify when a dose can be administered in a treatment schedule. Alternatively, a placebo dose or a calcium dietary supplement, similar or different to the dose of the pharmaceutical composition, may be included to provide a package in which the dose is received daily. Optionally in combination with such a container, may be a notice prescribed by a governmental agency regulating the manufacture, use or sale of pharmaceuticals, which notice reflects approval by the agency of manufacture, use or sale for human administration.

Equivalents of the same

The following representative examples are intended to aid in the description of the invention and are not intended to, or should not be construed as, limiting the scope of the invention. Indeed, various modifications of the invention and many additional embodiments thereof, in addition to those shown and described herein, will become apparent to those skilled in the art from the entire contents of these documents, including the examples that follow and references to the scientific and patent documents cited herein. It should also be understood that the contents of those references cited are incorporated herein by reference to help explain the prior art.

The following examples contain important additional information, exemplification and guidance which can be used to practice the invention in its various embodiments and equivalents thereof.

Examples of the use of

The compounds of the present invention and their preparation may be further understood by the following examples which illustrate some of the methods of making or using these compounds. It should be understood, however, that these examples do not limit the present invention. Variants of the invention, whether now known or further developed, are considered to be within the scope of the invention as described and claimed herein.

1) General description of the synthetic methods:

the practitioner has a very tangible literature on macrolide chemistry for citation, in combination with the information contained herein, as a guide in synthetic strategies, protecting groups, and other materials and methods for synthesizing the compounds of the invention.

Various references cited herein provide useful background information on the preparation of compounds similar to those described herein or of the invention or related intermediates, as well as information on the formulation, use, and administration of such compounds that may be of interest.

In addition, practitioners are guided by reference to the specific guidance and examples provided in these references for various exemplary compounds and intermediates thereof.

The compounds of the present invention and their preparation may be further understood by the following examples which illustrate some of the methods of making or using these compounds. It should be understood, however, that these examples do not limit the present invention. Variants of the invention, whether now known or further developed, are considered to be within the scope of the invention as described herein and claimed below.

In accordance with the present invention, any available technique may be used to produce or prepare the compounds of the present invention or compositions comprising them. For example, a variety of liquid phase synthesis methods may be used, such as those specifically discussed below. Alternatively or additionally, the compounds of the invention can be prepared using a variety of combinatorial techniques, parallel synthesis, and/or solid phase synthetic methods known in the art.

As described below, it is understood that various compounds of the invention can be synthesized according to the methods described herein. The starting materials and reagents for preparing these compounds are either obtained from commercial suppliers such as Aldrich Chemical Company (Milwaukee, Wis.), Bachem (Torrance, CA), Sigma (St. Louis, Mo.), or prepared by methods well known to those skilled in the art as described in the following references: fieser and Fieser, 1991, "Reagents for organic Synthesis", Vol.1-17, John Wiley and Sons, New York, NY, 1991; rodd 1989 "Chemistry of Carbon Compounds", Vol.1-5 and supplementary Vol.Elsevier Science Publishers, 1989; "Organic Reactions", Vol.1-40, John Wiley and Sons, New York, NY, 1991; march2001, "Advanced Organic Chemistry", 5 th edition, John Wiley and Sons, New York, NY; and Larock 1990, "Comprehensive Organic Transformations: A Guide to functional groups Preparations", 2 nd edition, VCH Publishers. These schemes are merely illustrative of some of the methods by which the compounds of the present invention can be synthesized, and various modifications to these schemes can be made and suggested to one skilled in the art having regard to this disclosure.

The starting materials, intermediates, and compounds of the present invention can be isolated and purified using conventional methods, including filtration, distillation, crystallization, chromatography, and the like. They can be characterized by conventional methods, including physical constants and spectral data.

General reaction methods:

unless specifically mentioned, the reaction mixture was stirred using a magnetically driven stir bar. The inert atmosphere refers to anhydrous argon or anhydrous nitrogen. The reaction is monitored by thin layer chromatography, by proton Nuclear Magnetic Resonance (NMR) or by High Pressure Liquid Chromatography (HPLC) of a suitably treated sample of the reaction mixture.

General post-treatment methods:

unless specifically mentioned, the reaction mixture was cooled to room temperature or below and then quenched with water or saturated aqueous ammonium chloride solution, if necessary. The desired product is partitioned between water and a suitable water immiscible solvent (e.g. ethyl acetate, dichloromethane, diethyl ether). The extract containing the desired product is suitably washed with water, followed by a saturated saline solution. In the case where the extract containing the product is considered to contain residual oxidizing agent, the extract is washed with a 10% sodium sulfite solution in a saturated aqueous sodium bicarbonate solution prior to the above washing process. In the case where the extract containing the product is considered to contain residual acid, the extract is washed with a saturated aqueous sodium bicarbonate solution (excluding those cases where the desired product itself has acidic properties) before the above washing process is carried out. In the case where the extract containing the product is considered to contain residual alkali, the extract is washed with a 10% aqueous citric acid solution before the above washing process is performed (excluding those cases where the desired product itself has alkalinity). After washing, the extract containing the desired product was dried over anhydrous magnesium sulfate, and then filtered. The crude product is then isolated by removal of the solvent by rotary evaporation at a suitable temperature (typically below 45 ℃) under reduced pressure.

General purification methods:

unless specifically mentioned, chromatographic purification refers to flash column chromatography on silica, using a single solvent or a mixed solvent as eluent. The eluates containing the desired product, suitably purified, are combined and concentrated to constant quality at a suitable temperature (typically below 45 ℃) under reduced pressure. The final compound was dissolved in 50% aqueous acetonitrile, filtered and transferred to a vial, then lyophilized under high vacuum and finally submitted for biological testing.

1) Synthesis of exemplary compounds:

unless otherwise indicated, starting materials are either commercially available or readily available by laboratory synthesis to those skilled in the art. The following generally describes methods and general guidance for the synthesis of compounds as generally described herein and in the subclasses and classes herein. In addition, synthetic guidance can be found in published PCT applications WO 99/49856 and WO02/059114, the entire contents of which are incorporated herein by reference.

Detailed Description

Example 1

This example describes the synthesis of the following compounds

Prepared according to scheme 1A and following procedures.

Reaction scheme 1A

a) A solution of 3-methoxyphenylethylamine (0.2mol) and formaldehyde (0.22mol) in aqueous HCl (20%, 500mL) was heated at 80 ℃ for 4 hours. The reaction was then concentrated to dryness, and the residue was dissolved in hydrobromic acid (40% aqueous, 500mL) and refluxed for 24 hours. The reaction was concentrated to give a tan solid, which was used without purification. To the residue were added water (200mL) and tetrahydrofuran ("THF") (300mL) and to the resulting mixture was added sodium carbonate (solid, 0.5mol) very carefully followed by di-tert-butyl dicarbonate (0.3 mol). After 15 hours at room temperature, the reaction was extracted with ethyl acetate (1L), and the organic extract was washed with saturated potassium dihydrogen phosphate and brine, dried over anhydrous magnesium sulfate and filtered.

After the filtrate was concentrated, the residue was dissolved in dichloromethane ("DCM", 100mL) and acetic acid (500mL) and sulfuryl chloride (0.6mol) were slowly added thereto. After stirring the reaction mixture at room temperature for 24 hours, the reaction was concentrated to dryness and additionally dried under high vacuum for 2 hours. The crude product was used in the next step without further purification. The crude product was dissolved in water/THF (200mL/400mL) and sodium carbonate (0.5mol) was added carefully and slowly with good stirring, followed by di-tert-butyl dicarbonate (0.3 mol). After stirring the reaction mixture for 12 hours, the reaction was carefully neutralized with phosphoric acid (2M) to a pH of about 7. The resulting mixture was extracted with ethyl acetate (500mL x2), and the combined extracts were washed with water and brine, dried over anhydrous magnesium sulfate, filtered, and concentrated. The crude solid was recrystallized from ethyl acetate and hexane (ratio of about 1: 2) to give a white solid. Concentrating the mother liquor, purifying with column, and purifying with 0-10% ethyl acetate4:1 hexane: dichloromethane. The combined yield was 14.5g (23% from commercial 3-methoxyphenethylamine). MS (API-ES)⁺) M/z 262,264,266(M + H-tert-butyl)⁺)。

The product obtained above was dissolved in DCM (100mL) and pyridine (50 mL). The resulting solution was cooled to-40 ℃ and trifluoromethanesulfonic anhydride (51mmol) was slowly added thereto. After the reaction mixture was gradually warmed to room temperature over 4 hours, the reaction mixture was partitioned between ethyl acetate (500mL) and water (100mL), and the organic layer was washed with water (100mL, twice) and brine (50mL), dried over anhydrous magnesium sulfate, filtered, and concentrated. The residue was purified by column chromatography eluting with 0-5% ethyl acetate in 5:1 hexanes: DCM to give the corresponding triflate (9.73g, 48% yield).

10mmol of triflate, 1.0mmol of 1, 3-diphenylphosphinopropane ("dppp") and 40mmol of diisopropylethylamine ("DIEA") in 100mL of anhydrous dimethylformamide ("DMF") and 50mL of anhydrous CH₃The mixture in OH was purged with CO for 15 minutes, then 1.0mmol of Pd (OAc) was added under a CO atmosphere₂. Subsequently, the resulting mixture was stirred under a CO atmosphere at 70 ℃ overnight. The solvent was removed and the residue was purified by column chromatography using EtOAc/hexanes-1/4 (v/v) as eluent to give compound 1.1 in 56% yield. ESI-MS (M/z): (M)⁺)+Na 382.1；¹H NMR(CD₃OD,400MHz):7.32(s,1H),4.60(s,2H),3.95(s,3H),3.69(m,2H),2.84(m,2H),1.50(s,9H)ppm。

b) A mixture of 1.1(5mmol) and 30mmol of LiI in 20mL of pyridine was refluxed overnight. The solvent was removed and the residue was dissolved in EtOAc. The resulting solution was then saturated with NH₄Aqueous Cl solution and anhydrous Na₂SO₄And (5) drying. The solvent was removed and the residue was dried in vacuo to give compound 1.3 in quantitative yield. The crude product was carried on to the next step without further purification. ESI-MS (M/z): (M-tBu +1), 290.

c) A solution of Boc-Dap-OH (10mmol) in methanol (30mL) was treated with trimethylsilyldiazomethane until the color remained pale yellow for 10 seconds. The mixture was concentrated and the residue was dissolved in DCM (30 mL). To the solution was added triethylamine (20mmol) followed by 3-thienylformyl chloride (11 mmol). After 0.5 h at room temperature, the reaction was filtered through silica gel and concentrated. The residue was purified by column chromatography eluting with 10-50% ethyl acetate in hexane. The product thus obtained was dissolved in DCM (10mL) and treated with HCl (4M in dioxane, 10 mL). After 5 hours, the reaction was concentrated to give the title compound (60-80% overall yield).

d) 1.2(4mmol), 1.3(4.4mmol), 5.0mmol of o- (7-azabenzotriazol-1-yl) -N, N, N ', N' -tetramethyluronium fluorophosphate ("HATU") and 20mmol of Et₃A mixture of N in 20mL of DMF was stirred at room temperature overnight. The solvent was removed and the residue was purified by column chromatography using CH₂Cl₂EtOAc 6/4(v/v) as eluent gave compound 1.4 in 60% yield. ESI-MS (M/z): (M +1) 556.1.

e) 2mmol of 1.4 in 9mL of TFA and 3mL of CH₂Cl₂The solution in (a) was stirred at room temperature for 6 hours. The solvent was removed and the residue was taken up with saturated NaHCO₃And (5) diluting the aqueous solution. The mixture was extracted 3 times with EtOAc. Then using anhydrous Na for extracting the extract₂SO₄And (5) drying. The solvent was removed and the residue was dried in vacuo to give compound 1.5, which was used without further purification. ESI-MS (M/z): (M +1)456.1.

f) Intermediate compound 1.11 was prepared according to scheme 1B and following procedure.

Reaction scheme 1B

To 100mmol of commercially available 6-hydroxy- [2H ] at room temperature]-benzofuran-3-one (compound 1.8) and 150mmol of imidazole in 300mL of anhydrous DMF 110mmol of tert-butyldimethylsilyl chloride ("TBDMSCl") were added, the resulting mixture was stirred overnight at room temperature, the solvent was removed, and the residue was taken up in 100mmolmL EtOAc dilution with saturated NH₄Washing with aqueous Cl solution and anhydrous Na₂SO₄Drying, removal of the solvent and purification of the residue gave the corresponding intermediate in 70% yield. ESI-MS (M/z): (M + H)⁺)265.1。

Dissolve the intermediate in 100mL of CH₃OH, 20mmol of NaBH was added at room temperature₄After stirring at room temperature for 12 hours, the reaction mixture was treated with 10mL of acetone, then 60mL of 4.0N HCl was added to the mixture, the mixture was stirred at room temperature overnight, the organic solvent was removed, the residue was extracted several times with EtOAc, then the extract was washed with brine, washed with anhydrous Na₂SO₄Drying, removal of the solvent and dissolution of the residue in 100mmol Et₃N and 180mL of anhydrous CH₂Cl₂66mmol of PhNTf were added at 0 deg.C₂The resulting mixture was stirred at rt overnight, the solvent was removed and the residue was purified to give compound 1.9 in 90% yield.¹HNMR(400MHz,CD₃Cl):7.75(d,J＝1.9Hz,1H),7.66(d,J＝8.5Hz,1H),7.50(s,1H),7.21(d,J＝8.5Hz,1H),6.85(d,J＝1.9Hz,1H)ppm。

50mmol of the compound 1.9, 2.5mmol of dppp (diphenylphosphine-1, 3-propane) and 2.5mmol of Pd (OAc)₂A mixture of 100mmol DIEA, 125mL anhydrous DMF, 125mL anhydrous MeOH was stirred at 65 ℃ overnight under an atmosphere of CO, the solvent was removed, and the residue was purified by column chromatography to give compound 1.10, 65% yield.¹H NMR(400MHz,CDCl₃):8.23(s,1H),7.99(d,J＝8.3Hz,1H),7.78(s,1H),7.65(d,J＝8.3Hz,1H),6.85(s,1H),3.97(s,3H)ppm；ESI-MS(m/z):(M+1)177.10。

20mmol of compound 1.10 and 80mmol of LiOH & H₂O in 60mL of THF and 15mL of H₂The mixture in O was stirred at room temperature for 1 hour, then 80mL of 1.0N aqueous HCl was added, the organic solvent was removed, the residue was diluted with 50mL of brine, then the mixture was extracted with EtOAc and the extract was extracted with anhydrous Na₂SO₄Drying, removal of the solvent and vacuum drying of the residue gave compound 1.11 in quantitative yield.¹H NMR(400MHz,CD₃OD):8.14(s,1H),7.92(m,2H),7.67(d,J＝8.5Hz,1H),6.92(s,1H)ppm；ESI-MS(m/z):(M+H⁺)163.1。

g) A mixture of 0.25mmol of compound 1.11 and 0.26mmol of HATU in 1mmol of DIEA and 2mL of DMF was stirred at room temperature for 30 minutes, then 0.22mmol of compound 1.5 in 1mL of DMF was added and the resulting mixture was stirred at 45 ℃ for 12 hours, the solvent was removed and the residue was purified to give compound 1.6 in 50-65% yield. Then, compound 1.6 was hydrolyzed with LiOH (1.0M aqueous solution, 0.5mL) in THF (3mL) for 2 hours, and then the reaction mixture was acidified with HCl (aqueous solution), extracted with ethyl acetate (50mL), dried over anhydrous magnesium sulfate, and concentrated to give compound 1 in quantitative yield.¹HNMR(400MHz,CD₃OD) 7.91(s,1H),7.75(d, J ═ 8.0Hz,1H),7.67(s,3H),7.36(d, J ═ 8.0Hz,1H),7.13(s,1H),6.96(s,1H),5.01(t, J ═ 6.8Hz,1H),4.68 and 4.89(m,2H),3.85(d, J ═ 6.8Hz,2H),3.70 and 4.02(m,2H),2.93(m,2H) ppm; ESI-MS (M/z): (M +1) 586.10.

Example 2

This example describes the synthesis of the following compounds,

it was prepared according to the procedure for example 1g, except that 4-chlorobenzoic acid was used instead of compound 1.11.¹HNMR(400MHz,CD₃OD) 7.64(m,2H),7.35-7.49(m,5H),7.11(s,1H),4.98(t, J ═ 8.0Hz,1H),4.63 and.88 (m,2H),3.83(d, J ═ 8.0Hz,2H),3.68 and 3.98(m,2H),2.89(m,2H) ppm; ESI-MS (M/z): (M +1) 579.90.

Example 3

This example describes the synthesis of the following compounds,

it is prepared according to scheme 2 and the following procedure.

Reaction scheme 2

a) To 10mmol of commercial 3.1 in 20mL MeOH and 20mL CH at 0 deg.C₂Cl₂To the solution in (1) was slowly added 20mmol of 2.0M TMSCHN in hexane₂The resulting mixture was stirred at room temperature for 30 minutes. The solvent was removed and the residue was dried in vacuo to give crude 3.2.

b) Then, compound 3.2 was reacted with 15mmol of isopropyl azide in 0.2mmol of CuI, 0.2mmol of Et₃In the presence of N at 50mL of CH₃Stirring overnight at room temperature in CN. The solvent was removed and the residue was purified by column chromatography to give compound 3.3, 55% yield. ESI-MS (M/z): (M +1) 313.20.

c) A mixture of 2mmol of compound 3.3 in 10mL of 4.0N HCl in dioxane was stirred at room temperature for 12 hours. The solvent was removed and the residue was dried in vacuo to give compound 3.4 in quantitative yield. ESI-MS (M/z): (M +1) 213.10.

d) A solution of 1.1(3.60g,10mmol) in DCM (20mL) at room temperature was treated with HCl in 1, 4-dioxane (4.0M,10 mL). After 2 hours, the reaction was concentrated to give compound 3.5 in quantitative yield.

e) Compound 3.5(10mmol) was mixed with EDC (2.11g,11mmol), N-dimethylaminopyridine ("DMAP",0.1g), triethylamine (2.02g) and compound 1.11(1.62g,10mmol) in dry DMF (50mL) and after 15 hours at room temperature the reaction mixture was diluted with ethyl acetate (200mL), washed with water (30mL,3 times), dried over anhydrous magnesium sulfate and filtered. The residue after concentration of the filtrate was purified by column chromatography eluting with 10-30% ethyl acetate in hexane to give the title compound (3.7g, 92%): ESI-MS (M/z): (M +1) 213.1.

f) Compound 3.7 was prepared according to example 1b, except that compound 3.6 was used instead of compound 1.1.

g) A mixture of 0.25mmol of compound 3.7 and 0.26mmol of HATU in 1mmol of DIEA and 2mL of DMF was stirred at room temperature for 30 minutes, then a solution of 0.22mmol of compound 3.4 in 1mL of DMF was added. The resulting mixture was stirred at 45 ℃ for 4 hours. The solvent was removed and the residue was purified to give an intermediate ester which was then treated with LiOH in THF and water to give the desired compound 3 in quantitative yield.¹H NMR(400MHz,CD₃OD)7.90(s,2H),7.74(m,1H),7.64(s,1H),7.34(m,1H),6.93(s,1H),5.03(m,1H),4.82(m,1H),4.65 and 4.88(m,2H),3.72 and 3.97(m,2H),3.40(m,1H),3.18(m,1H),2.90(m,2H),1.55(m,6H) ppm; ESI-MS (M/z): (M +1) 570.1.

Example 4

This example describes the synthesis of the following compounds,

it is prepared according to scheme 3 and the following procedure.

Reaction scheme 3

a) Boc-DAP-OH (0.2g,1.0mmol), 2, 4-dichloropyrimidine (0.29g,2.0mmol), diisopropylethylamine (0.51mL,2.9mmol) were heated to 75 ℃ in ethanol (5mL) for 14 hours. The reaction mixture was cooled to room temperature and the solvent was evaporated under reduced pressure. The crude 4.1 obtained is sufficiently pure to be used for the subsequent chemical transformations.

b) The crude residue 4.1(0.31g,1.0mmol) was dissolved in 9:1 benzene: methanol (5mL), trimethylsilyldiazomethane (1.0mL,2.0M in hexanes) was slowly added to the stirred reaction mixture and stirred for an additional 1 hour, and the solvent was evaporated under reduced pressure to give a crude residue as an oil. Purification by silica gel column chromatography eluting with 50% ethyl acetate in hexane afforded pure 4.2(0.21g, 65%).

c) Compound 4.2(0.21g,0.6mmol) was dissolved in dichloromethane (5 mL). Trifluoroacetic acid (2.5mL) was added, the reaction stirred for 1 hour, and the resulting reaction mixture was concentrated to remove excess trifluoroacetic acid to give amine 4.3 in quantitative yield.

d) Compound 4.4 was prepared according to example 3d-f, except that 4-chlorobenzoic acid was used instead of compound 1.11.

e) Compound 4 was prepared according to example 3g, except that compound 4.4 was used instead of compound 3.7, and compound 4.3 was used instead of compound 3.4.

Example 5

This example describes the synthesis of the following compounds,

it is prepared according to scheme 4 and the following procedure.

Reaction scheme 4

a) To a solution of methanesulfonamide (1.01g,10.7mmol) in 15mL of DMF was added 20M aqueous NaOH (0.68mL,13.6mmol) to give a white precipitate. The solution was cooled to 0 deg.C, carbon disulfide (0.4mL,6.63mmol) was slowly added, and stirred at 0 deg.C for 15 minutes. An additional 20M aqueous NaOH solution (0) was added.32mL,6.4mmol) and carbon disulphide (0.2mL,3.31mmol), the reaction is stirred at 0 ℃ for 20 minutes and then warmed to room temperature, all precipitates have become solution by 30 minutes, the reaction mixture is cooled to 0 ℃, methyl iodide (1.33mL,21.364mmol) is added, the reaction is stirred at 0 ℃ for 20 minutes and at room temperature for 1.5 hours, 20mL of water is added to the reaction and extracted 5 times with ethyl acetate. The combined organic extracts were extracted with MgSO₄Drying and concentration to dryness recrystallized from hot ethyl acetate and hexane to give 1.44g of compound 5.1. ESI-MS (M/z): (M + H)⁺)200.0。

b) To a solution of Boc-Dap-OH (109mg,0.53mmol) and compound 5.1(125.7mg,0.632mmol) in 5mL ethanol was added 1.0M aqueous NaOH (0.8mL,0.8 mmol). The reaction was stirred until complete conversion, then concentrated to dryness, the residue was dissolved in water, washed three times with ether, and the aqueous layer was acidified to pH 1 with 2.0M phosphoric acid and extracted 4 times with ethyl acetate. The combined ethyl acetate extracts were extracted with MgSO₄Dried and concentrated to dryness to give 160.4mg of compound 5.2. ESI-MS (M/z): (M + Na)⁺)378.0。

c) To a solution of compound 5.2(160.4mg,0.452mmol) in 1:1 dichloromethane: methanol was added trimethylsilyldiazomethane, which is a 2.0M solution in ether, (0.4mL,0.8 mmol). The reaction was stirred at room temperature until complete conversion to the methyl ester, then concentrated to dryness. The product was purified by flash chromatography to give 146.6mg of compound 5.3. ESI-MS (M/z): (M + Na)⁺)270.0；¹H NMR (400MHz, chloroform-d). 1.46(s,9H),2.42(s,3H),3.02(s,3H),3.63(m,1H),3.75(m,1H),3.82(s,3H),4.51(m, 1H).

d) To a solution of compound 5.3(146.6mg,0.40mmol) in 2mL of methanol was added ammonia, 7N in methanol (0.6mL,4.2 mmol). The mixture was cooled to 0 ℃, a solution of silver nitrate (75.5mg,0.444mmol) in 0.4mL acetonitrile was added dropwise, the reaction stirred and brought to room temperature, the solvent was removed after 3 hours, the residue was suspended in ethyl acetate, filtered through celite, the filtrate was concentrated to dryness and dissolved again in dichloromethane to which HCl,4.0M in dioxane (0.5mL,2.0mmol) was added. The solution was stirred at room temperature for 8 hours and concentratedTo dryness, compound 5.4 was obtained as the HCl salt. ESI-MS (M/z): (M + H)⁺)239。

e) Compound 5 was prepared according to example 3g, except that compound 4.1 was used instead of 3.7, and compound 5.4 was used instead of compound 3.4. ESI-MS (M/z): (M + H)⁺)590.0。

Example 6

This example describes the synthesis of the following compounds,

it is prepared according to scheme 5 and the following procedure.

Reaction scheme 5

a) A solution of (L) -Boc-Dap-OH (10mmol), dimethyl N-cyano dithioimine carboxylic acid (10mmol) and DIEA (30mmol) in ethanol was stirred at room temperature for 1 hour, then pyrrolidine (20mmol) was added and the reaction heated at 65 ℃ for 10 hours, the reaction mixture was diluted with ethyl acetate (150mL) and saturated NaH was used₂PO₄(50mL, pH of aqueous layer 4-6), water (50mL) and brine. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated, and the crude product was dissolved in 4:1 DCM: MeOH, cooled to 0 deg.C, and trimethylsilyldiazomethane was added thereto until it remained yellow for 30 seconds. The solution was concentrated and the residue was purified by column chromatography eluting with 30-100% ethyl acetate in hexanes to give the title compound in 20-30% yield. MS (M/z) 240 (M-99).

b) Compound 6.1(72mg,0.20mmol) was treated with HCl (4M in dioxane) in DCM (1mL) for 2 h. The reaction mixture was concentrated to give compound 6.2.

c) Compound 6 was prepared according to example 3g, except that compound 6.2 was used instead of compound 3.4.¹HNMR(400MHz,CDCl₃)7.77(s,1H),7.63(m,3H),7.37(d,1H),6.85(s,1H),6.32(d,1H),4.68(s,2H),4.31(m,1H),3.85(m,2H),3.60(m,4H),2.88(m,2H),1.89(m,4H)ppm；ESI-MS(m/z):(M+1)597.1。

Example 7

This example describes the synthesis of the following compounds

Prepared according to scheme 6 and following procedures.

Reaction scheme 6

a) To a solution of H-2-mercapto-His-OH (598.8mg,3.2mmol) in 5mL of methanol was added 0.26mL of sulfuric acid. The mixture was stirred at room temperature for 8h, additional sulfuric acid (0.1mL,0.6mmol) was added, the reaction stirred at 50 ℃ for 4.5 h, sodium carbonate (850.3mg,8.02mmol) was slowly added, the reaction dried by rotary evaporation to remove the bulk of the methanol, 6mL of THF and 3mL of water were added, along with sodium carbonate (0.851g,8.03mmol) and Boc₂O (696.5mg,3.19 mmol). The reaction was stirred at room temperature for 1 hour, diluted with ethyl acetate, washed with water and brine, and the organic layer was MgSO₄Dried and concentrated to dryness. Purification by flash chromatography gave 283.5mg of compound 7.1. ESI-MS (M/z): (M + H)⁺)316.1。

b) Compound 7.1(283.5mg,0.90mmol) and 3-chloroperbenzoic acid (568.4mg,2.53mmol) were dissolved in 6mL of dichloromethane and stirred at room temperature for 1.5 h, the reaction diluted with dichloromethane and poured into 1.0M potassium carbonate. The aqueous layer was adjusted to neutral pH with 2.0M phosphoric acid, thenExtracting with dichloromethane for 3 times, mixing the organic extracts, and collecting the organic extract with MgSO₄Dried and concentrated to dryness. Purification by flash chromatography gave 240.8mg of compound 7.2. ESI-MS (M/z) 370.1(M + Na)⁺),248.1(M-Boc+H⁺)；¹H NMR(400MHz,CDCl₃)1.43(s,9H),3.14(m,2H),3.26(s,3H),3.74(s,3H),4.61(m,1H),7.02(s,1H)。

c) To a solution of compound 7.2(240.8mg,0.6931mmol) in dichloromethane was added HCl,4.0M in dioxane (1.0mL,4.0 mmol). The reaction was stirred at room temperature for 1.5 hours, then concentrated to dryness to give compound 7.3 as the hydrochloride salt. ESI-MS (M/z): (M + H)⁺)248。

d) Compound 7 was prepared according to example 3g, except that compound 4.1 was used instead of compound 3.7, and compound 7.3 was used instead of compound 3.4. ESI-MS (M/z): (M + H)⁺)599.0；¹H NMR(400MHz,CDCl₃)2.88(s,2H),3.10(m,1H),3.21(s,3H),3.34(m,1H),3.67(1H),3.97(1H),4.63(1H),4.85(1H),5.04(dd,1H),7.19(s,1H),7.49(m,5H)。

Example 8

This example describes the synthesis of the following compounds

Prepared according to scheme 7A and following procedures.

Reaction scheme 7A

a) Three-necked flask containing 32mmol of LiCl was placed in a vacuumFlame burning with gas lamp under the air, then burning with N₂And (5) purging. This process was repeated 3 times to dry the LiCl. At room temperature under N₂To the flask was added 10mmol of (R, R) - (-) -pseudoephedrine glycinamide hydrate (pseudo-ephedrine glycinamide hydrate, A.G.Myers et al, J.Org.chem., 64:3322-3327(1999)) and 30mL of anhydrous THF under an atmosphere. The suspension was then treated with 31mmol of LiHMDS (1.0M in THF) at 0 ℃ for 1 hour with stirring, and then a solution of 10mmol of 3-methylthiobenzyl chloride (S.Laufer et al, J.Med.chem., 45:2733-2740(2002)) in 5mL of anhydrous THF was added. The resulting mixture was stirred at 0 ℃ overnight, and 10mL of water was added to quench the reaction. The solvent was removed and the residue was diluted with 50mL of water. CH for the mixture₂Cl₂Extracting for 3 times, mixing organic extracts, and adding anhydrous Na₂SO₄And (5) drying. The solvent was then removed and the residue was purified to give the desired alkylated intermediate in 70% yield. ESI-MS (M/z): (M + H)⁺)359.2。

A mixture of 5mmol of the alkylated intermediate in 12mL of 1.0N NaOH was refluxed until the starting material was consumed, the mixture was diluted with 20mL of water and CH₂Cl₂Extracting for 3 times, and then aqueous phase and 6mmol (Boc)₂O and 12mmol NaHCO₃Stirring in 30mL of 1, 4-dioxane for 15 hours, removing the organic solvent, diluting the residue with 30mL of water, diluting with CH₂Cl₂And (4) extracting. The aqueous phase was then treated with solid citric acid to adjust the pH to 4.0 and then extracted with EtOAc 3 times and the organic extracts were extracted with anhydrous Na₂SO₄And (5) drying. The solvent was removed and the residue was dried in vacuo to give compound 8.2 in quantitative yield. ESI-MS (M/z): (M + H)⁺)334.10。

b) To 5mmol of compound 8.2 in 10mL of CH at 0 deg.C₃OH and 10mL of CH₂Cl₂To the mixture of (2.0M in hexane) was added 10mmol of (trimethylsilyl) diazomethane, and the resulting mixture was stirred at room temperature for 30 minutes. The solvent was removed and the residue was dried in vacuo to give the desired ester in quantitative yield. ESI-MS (M/z): (M + H)⁺)348.10。

To the ester at 20mL CH at room temperature₃To a solution of OH and 2mL of water was added 12mmolThe resulting suspension was stirred at room temperature for 15 h, the reaction mixture was concentrated and diluted with 50mL of EtOAc, washed with water, and dried over anhydrous Na₂SO₄And (5) drying. The solvent was removed and the residue was dried in vacuo to give compound 8.3 in quantitative yield.¹H NMR(400MHz,CD₃Cl):7.82(m,1H),7.69(s,1H),7.50(m,1H),7.43(m,1H),5.03(m,1H),4.62(m,1H),3.74(s,3H),3.26(m,1H),3.09(m,1H),3.03(s,3H),1.39(s,9H)ppm；ESI-MS(m/z):(M–tBoc+H⁺)258.1。

c) A mixture of 2mmol of compound 8.3 in 10mL of 4.0N HCl in dioxane was stirred at room temperature for 15 h. The solvent was removed and the residue was dried in vacuo to give compound 8.4 as the hydrochloride salt in quantitative yield.¹H NMR(400MHz,CD₃OD):7.95(m,1H),7.87(s,1H),7.64(m,2H),4.44(t,J＝6.85Hz,1H),3.82(s,3H),3.41(m,1H),3.29(m,1H),3.13(s,3H)ppm；ESI-MS(m/z):(M+H⁺)258.10。

d) Compound 8 was prepared according to example 3g, except that compound 4.1 was used instead of compound 3.7, and compound 8.4 was used instead of compound 3.4.¹H NMR(400MHz,CD₃OD)7.92(s,1H),7.81 (d, J ═ 7.92Hz,1H),7.68(d, J ═ 7.83Hz),7.56(t, J ═ 7.83Hz,1H),7.30-7.49(m,5H),5.06(m,1H),4.60 and 4.83(m,2H),3.95 and 3.66(m,2H),3.44(d, J ═ 13.44Hz,1H),3.14(m,1H),3.08(s,3H),2.85(m,2H) ppm; ESI-MS (M/z): (M + H)⁺)609.05。

e) Compound 8.4 can also be prepared according to scheme 7B and the following procedures.

Reaction scheme 7B

8.5(1.0mmoL), methyl iodide (1.2 mmoL)) And a mixture of potassium carbonate (2mmoL) in 20mL of acetone was heated at 50 ℃ for 3 hours. The solvent of the reaction mixture was removed under reduced pressure and the residue was partitioned between EtOAc and water. The aqueous solution was extracted with EtOAc and the combined organic solutions were washed with brine, Na₂SO₄Dried, filtered and concentrated. The crude product was a white solid (yield 98%), which was used directly in the next process without purification.

LiAlH was slowly added to a solution of the above-prepared compound (1mmoL) in 4mL of THF at 0 deg.C₄(1.1 mmoL). The reaction mixture was warmed to room temperature and stirred for 1 hour, and water, a 15% aqueous NaOH solution and water were sequentially added to the reaction and stirred vigorously. Filtration and evaporation of the filtrate gave crude 8.6 (92% yield). No purification is required.¹H NMR(400MHz,CDCl₃).3.05(s,3H),4.75(s,2H),7.53(t,J＝7.58Hz,1H),7.62(d,J＝7.34Hz,1H),7.81(d,J＝7.83Hz,1H),7.93(s,1H)。

At room temperature under N₂Next, solid ammonium tetrapropylperruthenate ("TPAP",0.05mmoL) was added in one portion to a stirred mixture of compound 8.6(1mmoL), 4-methylmorpholine N-oxide ("NMO"; 1.5mmoL) and powdered 4A molecular sieve (equivalent to the weight of NMO) in 5mL of DCM. The reaction mixture was stirred at room temperature for 1 hour, then filtered through a short pad of silica gel eluting with DCM and AcOEt (1: 1). The filtrate was concentrated and the residue was purified by chromatography (SiO)₂AcOEt/hexane 2:1) to give compound 8.7 (yield 72%).¹H NMR(400MHz,CDCl₃)3.14(s,3H),7.81(t,J＝7.58Hz,1H),8.21(t,J＝9.05Hz,2H),8.46(s,1H),10.12(s,1H)ppm。

N, N, N ', N', -tetramethylguanidine ("TMG"; 1.05mmole) was slowly added to a solution of (d, l) -Cbz- α -phosphonoglycine trimethyl ester (1.1mmole) in 4mL of DCM at room temperature, after 15 minutes, the mixture was cooled to-30 deg.C, compound 8.7(1mmole) was added dropwise, the mixture was held at-30 deg.C for 20 minutes and slowly warmed to 0 deg.C, the solution was diluted with AcOEt and sequentially with 1N NaHSO₄And brine washing. Drying of the solution (Na)₂SO₄) The solvent was evaporated to give the crude product. The crude product was purified by chromatography (SiO2, AcOEt/hexane/DCM 3:3:1) to afford the productProduct 8.8 (yield 72%).¹H NMR(400MHz,CDCl₃)2.97(s,3H),3.86(s,3H),5.08(s,2H),6.78(s,1H),7.34(d, J ═ 6.36Hz,5H),7.50(t, J ═ 7.83Hz,1H),7.72(d, J ═ 7.34Hz,1H),7.85(d, J ═ 7.34Hz,1H),8.04(s, 1H). The olefinic proton of the minor trans isomer was at 7.19ppm (s, 1H). ESI-MS (M/z): (M + H)⁺)346。

To a solution of the chiral catalyst, (+) -bis ((2S,5S) -2, 5-dimethylphospano) benzene (cyclooctadiene) rhodium (I) tetrafluoroborate (0.01mmole), in 8.8(1mmole) MeOH (20mL, pre-charged with nitrogen) in a glass pressure vessel was added. Then the reactor is used with H₂Pressurisation to 40psi and continued shaking at room temperature for 17 hours, the solvent was evaporated, the residue was dissolved in AcOEt and passed through SiO₂The pad was filtered with AcOEt. The filtrate was evaporated to give crude 8.9. (yield, 72%).¹H NMR(400MHz,CDCl₃)2.98(s,3H),3.13(dd,J＝13.69,6.36Hz,1H),3.29(m,1H),3.76(s,3H),4.69(m,1H),5.06(m,2H),5.44(d,J＝6.85Hz,1H),7.31(m,5H),7.41(d,J＝7.34Hz,1H,7.47(t,J＝7.83Hz,1H),7.72(s,1H),7.82(d,J＝7.34Hz,1H)。ESI-MS(m/z):(M+H⁺)348。

Hydrogenation of compound 8.9 using hydrogen balloon (Pd/C, MeOH) gave compound 8.4 (98% yield). ESI-MS (M/z): (M + H)⁺)258。

Example 9

This example describes the synthesis of the following compounds

It is prepared according to scheme 8 and the following procedure.

Reaction scheme 8

a) Compound 9.1 was prepared according to examples 1d-e, except that compound 8.4 was used instead of compound 1.3.

b) Compound 9 was prepared according to example 3g, except that compound 9.1 was used instead of compound 3.7 and 2-hydroxycinnamic acid was used instead of compound 3.4.¹H NMR(400MHz,CD₃OD)7.92(m,2H),7.81(m,1H),7.68(s,1H),7.58(m,2H),7.28(m,2H),7.18(m,1H),6.84(m,2H),5.07(m,1H),4.80 and 4.88(m,2H),3.96(m,2H),3.42(m,1H),3.15(m,1H),3.08(s,3H),2.71-2.91(m,2H) ppm; ESI-MS (M/z): (M + H)⁺)617.10。

Example 10

This example describes the synthesis of the following compounds

It was prepared according to example 3g, except using compound 9.1 instead of compound 3.7 and 2-fluorocinnamic acid instead of compound 3.4.¹H NMR(400MHz,CD₃OD)7.92(s,1H),7.81(m,3H),7.68(m,1H),7.57(m,1H),7.42(m,1H),7.29(m,2H),7.23(m,1H),7.16(m,1H),5.06(m,1H),4.81 and 4.88(m,2H),3.82 and 3.97(m,2H),3.44(m,1H),3.16(m,1H),3.08(s,3H),2.86 and 2.93(m,2H) ppm; ESI-MS (M/z): (M + H)⁺)619.10。

Example 11

This example describes the synthesis of the following compounds,

it was prepared according to example 3g, except that compound 9.1 was used instead of compound 3.7 and 6-indazole carboxylic acid was used instead of compound 3.4.¹H NMR(400MHz,CD₃OD)8.13(s,1H),7.93(m,3H),7.83(m,1H),7.70(m,1H),7.60(m,1H),7.20 and 7.34(m,1H),5.06(m,1H),4.64 and 4.88(m,2H),3.69 and 3.98(m,2H),3.40(m,1H),3.15(m,1H),3.08(s,3H),2.88(m,2H) ppm; ESI-MS (M/z): (M + H)⁺)615.15。

Example 12

This example describes the synthesis of the following compounds,

it was prepared according to example 3g, except using compound 9.1 instead of compound 3.7 and 2-indolecarboxylic acid instead of compound 3.4.¹H NMR(400MHz,CD₃OD)7.93(s,1H),7.83(m,2H),7.72(m,1H),7.83(m,1H),7.57(m,1H),7.45(m,1H),7.27(s,1H),7.20(m,1H),7.07(m,1H),6.93(s,1H),5.08(m,1H),4.88(m,2H),4.07(m,2H),3.46(m,1H),3.15(m,1H),3.08(s,3H),2.96(m,2H)ppm；ESI-MS(m/z):(M+H⁺)614.10。

Example 13

This example describes the synthesis of the following compounds,

it was prepared according to example 3g, except using compound 9.1 instead of compound 3.7 and 2-quinolinecarboxylic acid instead of compound 3.4.¹H NMR(400MHz,CD₃OD)8.51(m,1H),8.08(m,1H),8.01(m,1H),7.94(m,1H),7.84(m,2H),7.70(m,3H),7.58(m,1H),7.36 and 7.04(m,1H),5.07(m,1H),4.78 and 4.95(m,2H),3.79 and 4.08(m,2H),3.45(m,1H),3.17(m,1H),3.09(s,3H),2.97(m,2H) ppm; ESI-MS (M/z): (M + H)⁺)626.10。

Example 14

This example describes the synthesis of the following compounds,

it is prepared according to scheme 9 and the following procedure.

Reaction scheme 9

a) To a solution of 3-carboxybenzenesulfonyl chloride (3.54g,16mmol) in ethyl acetate (50mL) at 0 deg.C was added concentrated aqueous ammonia (2.5 mL). The reaction was neutralized with HCl in dioxane (20mL), diluted with ethyl acetate (100mL), dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated to give the title compound, which was used in the reaction without purification.

b) The crude compound 14.1 was dissolved in THF (50mL), borane (1.0M in THF, 50mL) was added over 20 minutes, and after stirring the reaction at room temperature for 15 minutes, the reaction was diluted with brine (20mL) and water (10mL) and then ethyl acetate (100 mL). The organic extract was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated to give the title compound, which was used in the reaction without purification.

c) To a solution of crude compound 14.2 in DCM (100mL) was added activated 4A molecular sieve powder (8g), pyridinium dichromate (7.55g,20 mmol). After the reaction was stirred at room temperature for 2 hours, the reaction mixture was filtered through silica gel (50g) and rinsed with ethyl acetate. The residue after concentrating the filtrate was purified by silica gel columnElution with 30-50% ethyl acetate in hexanes provided compound 14.3(477mg, 16%, 3 steps). ESI-MS (M/z): (M + H)⁺)186。

d) Compound 14.4 was prepared according to example 8e, except that compound 14.3 was used instead of compound 8.7. MS (ESI)⁺)m/z:260(M+H⁺)。

e) Compound 14 was prepared according to example 3g, except that compound 14.4 was used instead of compound 3.4.¹HNMR(400MHz,CD₃OD)7.89(s,1H),7.80(s,1H),7.75(m,2H),7.64(s,1H),7.57(d,1H),7.34(d,2H),6.93 9s,1H),5.00(m,1H),3.99(m,1H),3.73(m,1H),3.40(dd,1H),3.12(dd,1H),2.89(m,2H)ppm；ESI-MS(m/z):616(M+H⁺)。

Example 15

This example describes the synthesis of the following compounds

It is prepared according to scheme 10 and the following procedure.

Reaction scheme 10

a) To a solution of 0.2mol of furan in 200mL of anhydrous THF at-78 deg.C was added 0.2mol of n-BuLi (1.6M in hexanes) and the resulting solution was stirred at room temperature for 4 hours, then the mixture was cooled to-78 deg.C and stirred with 0.21mol of dimethyl sulfide at room temperature overnight, then 10mL of saturated NH was added₄Aqueous Cl solution. The mixture was concentrated at room temperature and the residue was taken up in 200mL of saturated NH₄Diluting with Cl aqueous solution and extracting with ether, washing the extract with brine and adding anhydrous Na₂SO₄And (5) drying. Removing the solventThe residue was distilled and the fractions at 135-140 ℃ C./760 mmHg were collected to give 15.1, 55% yield of the compound.¹H NMR(400MHz,CD₃Cl):7.50(s,1H),6.45(m,1H),6.39(s,1H),2.42(s,3H)ppm。

b) To a solution of 0.1mol of compound 15.1 in 100mL of anhydrous THF at-78 deg.C was added 0.1mol of n-BuLi (1.6M in hexanes) and the resulting solution was stirred at room temperature for 4 hours, then the mixture was cooled to-78 deg.C and treated with 0.12mol of anhydrous DMF and the mixture was stirred at room temperature overnight. The reaction was carried out by adding 10mL of saturated NH₄The mixture was quenched with aqueous Cl and concentrated. The residue was diluted with 100mL of saline and extracted with EtOAC. The extract was washed with brine and anhydrous Na₂SO₄And (5) drying. The solvent was removed and the residue was purified to give the title compound in 65% yield.¹H NMR(400MHz,CD₃Cl):9.52(s,1H),7.24(d,J＝3.4Hz,1H),6.42(d,J＝3.4Hz,1H),2.60(s,3H)ppm；ESI-MS(m/z):(M+H⁺)143.1。

c)50mmol of compound 15.2 and 120mmol of m-CPBA in 100mL of CH₂Cl₂The mixture of (1) was stirred at room temperature overnight. The mixture was treated with 150mL of CH₂Cl₂Diluting, and mixing with saturated NaHCO₃The aqueous solution was washed several times. Then the solution is treated with anhydrous Na₂SO₄Drying and concentration, the residue was purified to give compound 15.3 in 70% yield.¹H NMR(400MHz,CD₃Cl):9.83(s,1H),7.33(m,2H),3.27(s,3H)ppm；ESI-MS(m/z):(M+H⁺)175.0。

d) Compound 15.4 was prepared according to example 8e, except that compound 15.3 was used instead of 8.7. ESI-MS (M/z): (M + H)⁺)248.1。

e) Compound 15 was prepared according to the examples except that compound 15.4 was used instead of 3.4.¹H NMR(400MHz,CD₃OD)7.92(s,1H), 7.76(m,1H),7.67(s,1H),7.34(m,1H),7.13(s,1H),6.69(s,1H),6.49(s,1H),5.11(m,1H),4.73 and 4.88(m,2H),3.76 and 4.02(m,2H),3.46(m,1H),3.30(m,1H),3.17(s,3H),2.94(m,2H) ppm; ESI-MS (M/z): (M + H)⁺)605.05。

Example 16

This example describes the synthesis of the following compounds

It was prepared according to the following procedure.

To 0.2mmol of Compound 8.4 (example 8c or 8e) in 1mmol of Et at 0 deg.C₃N and 5mL of anhydrous CH₂Cl₂To the solution of (2, 6-dichlorobenzoyl chloride) was added 0.22mmol, the resulting mixture was stirred at room temperature for 12 hours, the solvent was removed, and the residue was dried in vacuo. The residue was then washed with 0.8mmol of LiOH H₂O in 2mL of THF and 0.5mL of H₂And (4) treating in O. After stirring at room temperature for 30 minutes, 1.0mL of 1.0N aqueous HCl was added to the reaction mixture. The organic solvent was removed and the residue was diluted with 10mL of brine. The mixture was extracted with EtOAc and the extract was extracted with anhydrous Na₂SO₄And (5) drying. The solvent was removed and the residue was dried in vacuo to afford the desired compound in 65% yield.¹H NMR(400MHz,CD₃OD)7.92(s,1H),7.82(d,J＝6.85Hz,1H),7.71(d,J＝6.85Hz,1H),7.56(t,J＝7.83Hz,1H),7.34(m,3H),5.08(dd,J＝9.78,4.89Hz,1H),3.45(dd,J＝14.67,4.89Hz,1H),3.14(dd,J＝14.67,9.78Hz,1H),3.08(s,3H)ppm；ESI-MS(m/z):(M+H⁺)416.00。

Example 17

This example describes the synthesis of the following compounds

It was prepared according to the following procedure.

Compound 6.1 (example 6a,0.2mmol) in DCM (1mL) was reacted with HCl in dioxane (4.0)M1 mL). After 1h, the solvent was evaporated and the residue and 1mmol Et were added at 0 deg.C₃N and 5mL of anhydrous CH₂Cl₂0.22mmol of 2, 6-dichlorobenzoyl chloride was added thereto, the resulting mixture was stirred at room temperature for 15 hours, the solvent was removed, and the residue was dried in vacuo. The residue was then taken up in 2mL of THF and 0.5mL of H₂0.8mmol of LiOH. H in O₂And (4) O treatment. After stirring at room temperature for 30 minutes, the reaction mixture was added to 1.0mL of 1.0N aqueous HCl. The organic solvent was removed and the residue was diluted with 10mL of brine. The mixture was extracted with EtOAc and the extract was extracted with anhydrous Na₂SO₄Drying, removal of the solvent and drying of the residue in vacuo afforded the desired compound.

Example 18

This example describes the synthesis of the following compounds

It is prepared according to scheme 11 and the following procedure.

Reaction scheme 11

a) 1 equivalent of LiAlH at-78 deg.C₄(1.0M in THF) was added to a solution of 1 equivalent of ethyl 6-indazolecarboxylate (Batt, D.G., J.Med.chem., 43:41-58(2000)) at 0 ℃. The reaction was stirred at-78 ℃ for an additional 30 minutes and then warmed to 0 ℃. Slowly add 1 equivalent of 1M aqueous NaOH. The resulting slurry was filtered through a short column of celite and washed with copious amounts of ethyl acetate. The combined organics were washed with MgSO₄Drying and vacuum concentratingThe alcohol 18.1 is obtained in sufficiently high purity to be used without further purification.

b) 1.1 equivalents of Dess-Martin periodinane (periodinane) were added to 1 equivalent of 18.1 in dichloromethane. After stirring the reaction at room temperature for 3 hours, the resulting precipitate was removed by filtration through a short pad of celite. The celite plug was washed with dichloromethane. The combined organics were concentrated to give aldehyde 18.2, which was sufficiently pure to be used without further purification.

c) 2.1 equivalents of ethylmagnesium bromide (0.5M in THF) were added to a pre-cooled solution of THF containing 18.2 at 0 deg.C. After 30 minutes, the reaction was allowed to warm to room temperature and stirred for an additional 2 hours. The resulting reaction mixture was diluted with ethyl acetate and washed with water. The organic layer was then washed with MgSO₄Drying, filtering and vacuum concentrating. The residue was then purified by silica gel column chromatography (using a gradient of ethyl acetate and hexane) to afford pure compound 18.3.

d) To 4-amino-2, 6-dichlorophenol (1 equivalent) in 3:2 THF/H₂NaHCO was added to the mixture in O₃(1.1 equiv.) and Boc₂O (1.1 equiv.) after stirring overnight, the reaction was extracted with ether and MgSO₄Drying, filtering and vacuum concentrating. The residue 18.4 was used without purification.

e) To a solution of phenol 18.4(1 eq) and 2, 6-lutidine (2.2 eq) in DCM at-78 deg.C was added trifluoromethanesulfonic anhydride (1.2 eq). After gradually warming the reaction mixture to room temperature overnight, the reaction mixture was diluted with ether, washed with water, and MgSO₄Drying, filtering and vacuum concentrating. The residue was then purified on a silica gel column (gradient elution with ethyl acetate and hexanes) to give pure compound 18.5.

f)10mmol of triflate 18.5,1.0mmol of dppp and 40mmol of DIEA in 100mL of anhydrous DMF and 50mL of anhydrous CH₃The mixture in OH was purged with CO for 15 minutes, then 1.0mmol of Pd (OAc) was added under a CO atmosphere₂. Subsequently, the resulting mixture was stirred at 70 ℃ under a CO atmosphereStirring overnight. The solvent was removed and the residue was purified by column chromatography eluting with 10-30% EtOAc in hexanes to give compound 18.6.

g)1 equivalent of Boc-aniline 18.6 was carefully dissolved in 6M H₂SO₄In aqueous solution, the mixture was then cooled to 0 ℃. To this was added slowly sodium nitrite (1.1eq in water) with vigorous stirring, followed by addition of sodium iodide (5 eq) over 1.5 hours. After the reaction was stirred at room temperature overnight, the reaction was diluted with ether, washed with water and MgSO₄Drying, filtering and vacuum concentrating. The residue was then purified on a silica gel column (gradient elution with ethyl acetate and hexanes) to give pure compound 18.7.

h) 1 equivalent of iodide 18.7, 1 equivalent of alkyne 18.3, 0.05 equivalent of CuI, 5 equivalents of triethylamine were dissolved in benzene and N was bubbled through the syringe needle into the solution₂The solution was degassed for a total of 15 minutes. 0.05 equivalent of PdCl was added₂(dppf). DCM. After 4 hours, the reaction was diluted with ethyl acetate and washed with water, brine. The organic layer was then washed with MgSO₄Drying, filtering and vacuum concentrating. The residue was then purified by silica gel column chromatography (using a gradient of ethyl acetate and hexane) to afford pure compound 18.8.

i) 1 equivalent of 18.8 was dissolved in MeOH and 5% Rh/Al was added₂O₃(20% by weight). Oxygen was removed from the flask under reduced pressure. The internal pressure was restored by adding hydrogen delivered using a hydrogen-filled balloon. The reaction was stirred under a hydrogen atmosphere for 14 hours. The reaction was filtered through a pad of celite and concentrated in vacuo. The residue was then purified by silica gel column chromatography (using a gradient of ethyl acetate and hexane) to afford pure compound 18.9.

j) 4 equivalents of LiI were added to 1 equivalent of compound 18.9 in pyridine. The reaction was refluxed for 14 hours and then allowed to cool to room temperature. The reaction was concentrated and the resulting residue partitioned between ethyl acetate and water. The aqueous layer was extracted with ethyl acetate. The combined organic layers were over MgSO₄Drying, filtering and concentrating. The residue is then passed over siliconPurification by gel column chromatography (gradient elution with ethyl acetate and methanol) afforded pure compound 18.10.

k) 1 equivalent of compound 18.10, 1 equivalent of compound 8.4 (example 8c or 8e), 3 equivalents of DIEA were dissolved in DMF. 1.1 equivalents of HATU were added. The reaction was stirred at room temperature for 14 hours. The reaction mixture was diluted with ethyl acetate and washed with water, brine. The combined organics were washed with MgSO₄Drying, filtering and concentrating. The residue was then purified by silica gel column chromatography (using a gradient of ethyl acetate and hexane) to give the pure intermediate ester. The ester was dissolved in methanol and 2 equivalents of 1MLiOH (aq) were added. Upon completion, excess solvent was removed under reduced pressure and the resulting acid was then purified by reverse phase HPLC to afford pure compound 18.

Example 19

This example describes the synthesis of the following compounds

Prepared by treating compound 18 (example 18) in DCM/TFA (1:1 ratio) with triethylsilane (10 equivalents). After LC-MS indicated complete consumption of the starting material, the reaction was concentrated and the residue was purified by reverse phase HPLC to give the title compound.

Example 19

This example describes the synthesis of the following compounds

Prepared according to scheme 12 and following procedures.

Reaction scheme 12

a) A solution of 1 equivalent of compound 18.7 (example 18g) in THF was treated with 1.0 equivalent of isopropyl magnesium bromide at-40 deg.C. After 0.5 h, DMF (5 equiv.) was added and the reaction warmed to room temperature overnight. The reaction mixture was diluted with ethyl acetate and washed with water, brine. The organic layer was then washed with MgSO₄Drying, filtering and vacuum concentrating. The residue was then purified by silica gel column chromatography (using a gradient of ethyl acetate and hexane) to afford pure compound 20.1.

b) To 20.1(1 eq) in THF was added 2.1 equivalents of acetylenic magnesium bromide (0.5M in THF) at-78 ℃. After the reaction was warmed to room temperature and stirred for an additional 2 hours, the resulting reaction mixture was diluted with ethyl acetate and washed with water. The organic layer was then washed with MgSO₄Drying, filtering and vacuum concentrating. The residue was then purified by silica gel column chromatography (using a gradient of ethyl acetate and hexane) to afford pure compound 20.2.

c) 1 equivalent of compound 20.2, 1 equivalent of 1-chloro-4-iodobenzene, 0.05 equivalent of CuI, 5 equivalents of triethylamine were dissolved in benzene and N was bubbled into the solution through the syringe needle₂The solution was degassed for a total of 15 minutes. 0.05 equivalent of PdCl was added₂(dppf). DCM. After 4 hours, the reaction was diluted with ethyl acetate and washed with water, brine. The organic layer was then washed with MgSO₄Drying, filtering and vacuum concentrating. The residue was then purified by silica gel column chromatography (using a gradient of ethyl acetate and hexane) to afford pure compound 20.3.

d) 1 equivalent of compound 20.3 was dissolved in MeOH. 5% Rh/Al addition₂O₃. Oxygen was removed from the flask under reduced pressure. The internal pressure was restored by adding hydrogen delivered using a hydrogen-filled balloon. The reaction was stirred under a hydrogen atmosphere for 14 hours. The reaction was filtered through a pad of celiteFiltered and concentrated in vacuo. The residue was then purified by silica gel column chromatography (using a gradient of ethyl acetate and hexane) to afford pure compound 20.4.

e) 4 equivalents of LiI were added to 1 equivalent of compound 20.4 in pyridine. The reaction was refluxed for 14 hours and then allowed to cool to room temperature. The reaction was concentrated and the resulting residue partitioned between ethyl acetate and water. The aqueous layer was extracted with ethyl acetate. The combined organic layers were over MgSO₄Drying, filtering and concentrating. The residue was then purified by silica gel column chromatography (using a gradient of ethyl acetate and methanol) to afford pure compound 20.5.

f)1 equivalent of compound 20.5, 1 equivalent of compound 8.4 (example 8c or 8e), 3 equivalents of DIEA were dissolved in DMF. 1.1 equivalents of HATU were added. The reaction was stirred at room temperature for 14 hours. The reaction mixture was diluted with ethyl acetate and washed with water, brine. The combined organics were washed with MgSO₄Drying, filtering and concentrating. The residue was then purified by silica gel column chromatography (using a gradient of ethyl acetate and hexane) to afford pure compound 20.6.

g)1 equivalent of compound 20.6 was dissolved in methanol, followed by the addition of 2 equivalents of 1M LiOH (aq). Upon completion, excess solvent was removed under reduced pressure and the resulting acid was purified by reverse phase HPLC and lyophilized to give pure powder compound 20.

Example 21

This example describes the synthesis of the following compounds

It is prepared according to scheme 13 and the following procedure.

Reaction scheme 13

a) A solution of bromine (461. mu.L, 9.00mmol) in methanol (6.0mL) was prepared at-78 ℃. The cold solution was added dropwise to a-78 deg.C mixture of KCN (1.85g,19.0mmol) in methanol (6.0mL) under nitrogen. After 20 minutes, a solution of pyrrole (0.624mL,9.00mmol) in methanol (20mL) was added. The mixture was brought to-40 ℃ and stirred for 0.5 h. It was then poured onto ice-water and extracted with ether (3 ×). The organic layers were combined and washed with saturated sodium thiosulfate and brine. The organic phase is then dried (Na)₂SO₄) And concentrated. The crude residue was purified by flash chromatography (0-50% ethyl acetate in hexanes) to give 734mg (66%) of compound 21.1 as a colorless oil, R_f0.24 (10% ethyl acetate in hexanes).¹H NMR(CDCl₃)8.68(br s,1H),7.03(s,1H),6.69(s,1H),6.32(s,1H)。ES(+)MS m/e＝125(M+H)⁺。

b) Aqueous sodium hydroxide (9.90mL,1M,9.90mmol) was added dropwise to a solution of 21.1(614mg,4.95mmol) and iodomethane (0.340mL,5.45mmol) in methanol (40mL) under nitrogen at-10 ℃. The mixture was allowed to warm to ambient temperature and stirred for 0.5 h. The excess sodium hydroxide was then quenched by the addition of dry ice. The mixture was diluted with brine and extracted with dichloromethane (3 ×). The combined organic layers were dried (Na)₂SO₄) And concentrated to give 504mg (90%) of compound 21.2 as a dark oil, R_f0.35 (10% ethyl acetate in hexanes).¹H NMR(CDCl₃)8.31(br s,1H),6.84(s,1H),6.38(s,1H),6.24(s,1H),2.36(s,3H)。

c) To a 0 ℃ solution of compound 21.2(100mg,0.884mmol) in methanol (4.0mL) was added sodium periodate (208mg,0.972mmol) in water (4.0mL) dropwise. The mixture was allowed to warm to room temperature and after 15 minutes LC/MS and TLC showed complete disappearance of compound 21.2. The mixture was then filtered, concentrated, and the residue partitioned between ethyl acetate and water. The aqueous layer was extracted twice with ethyl acetate and the combined organic phases were washed with brine and dried (Na)₂SO₄) And areConcentration gave 63mg (55%) of compound 21.3 as a dark oil, R_f0.25 (ethyl acetate).¹H NMR(CDCl₃)11.74(br s,1H),7.00(s,1H),6.65(s,1H),6.20(s,1H),3.03(s,3H)。ES(+)MS m/e＝130(M+H)⁺。

d) To a solution of compound 21.3(60mg,0.464mmol) in dichloromethane (2.0mL) was added trifluoroacetic acid (1.0 mL). After 15 min, LC/MS and TLC showed complete disappearance of compound 21.3. The solvent was removed in vacuo and the residue was dried under high vacuum to give 60mg (100%) of compound 21.4 as an oil, R_f0.11 (ethyl acetate).¹H NMR(CDCl₃)9.16(br s,1H),7.28(s,1H),6.94(s,1H),6.66(s,1H),3.02(s,3H)。ES(+)MS m/e＝130(M+H)⁺。

e) To compound 21.4(60mg,0.464mmol) and Cs₂CO₃(378mg,1.16mmol) of compound 21.5(Ferreira et al Tetrahedron Letters,39:9575 (1998); 140mg,0.464mmol) in acetonitrile was added. The resulting mixture was stirred at 60 ℃ until LC/MS and TLC showed complete consumption of starting material (-1 hour). The mixture was cooled to room temperature and diluted with ethyl acetate and washed with water and brine. The organic layer was dried (Na)₂SO₄) And concentrated. The crude residue was purified by flash chromatography (0-100% ethyl acetate in hexanes) to give 114mg (57%) of compound 21.6 as a viscous oil, R_f0.31 (ethyl acetate).¹H NMR(CDCl₃)7.05(s,1H),6.75(s,1H),6.50(s,1H),5.25(m,1H),4.65(m,1H),4.40(m,1H),3.79(s,3H),2.79(s,3H),1.47(s,18H)。ES(+)MS m/e＝275(M–Boc–t-Bu+2H)⁺。

f) To a solution of compound 21.6(114mg,0.265mmol) in dichloromethane (1.00mL) was added mCPBA (89.0mg,0.397mmol) dropwise. After 5 minutes of reaction at room temperature, LC/MS and TLC showed complete disappearance of compound 21.6. The mixture was filtered and concentrated. The crude residue was purified by flash chromatography (0-50% ethyl acetate in hexanes) to give 89.0mg (75%) of compound 21.7 as a colorless oil, R_f0.45 (50% ethyl acetate in hexanes).¹H NMR(CDCl₃)7.22(s,1H),6.69(s,1H),6.50(s,1H),5.27(m,1H),4.65(m,1H),4.45(m,1H),3.79(s,3H),305(s,3H),1.48(s,18H)。ES(+)MS m/e＝247(M–2Boc+3H)⁺。

g) To a solution of compound 21.7(89.0mg,0.0.199mmol) in dichloromethane (0.50mL) was added HCl (4.00mL,4.0M in dioxane). The resulting mixture was stirred at ambient temperature until LC/MS showed complete deprotection (-1 hour). The mixture was concentrated and the residue dried under high vacuum to give 53mg (100%) of compound 21.8 as a white powder. ES (+) MS M/e 247(M + H)⁺。

h) A mixture of compound 21.8(53.0mg,0.199mmol), compound 4.1 (example 4a,77.0mg,0.199mmol), HATU (79.0mg,0.209mmol), triethylamine (0.111mL,0.796mmol) in DMF (1.00mL) was stirred at room temperature overnight. The mixture was then diluted with ethyl acetate and washed with 1.0M aqueous HCl, saturated NaHCO₃And washing with saline water. The organic layer was dried (Na)₂SO₄) And concentrated. The crude residue was purified by flash chromatography (0-100% ethyl acetate in hexanes) to give 70.3mg (58%) of compound 21.9 as a white solid, R_f0.16 (75% ethyl acetate in hexanes).¹H NMR(CDCl₃)7.46-7.39(m,5H),7.28(s,1H),6.73(s,1H),6.69(br s,1H),6.48(s,1H),5.13(m,1H),4.83(br s,1H),4.56(m,1H),3.85(s,3H),3.70(m,1H),3.04(s,3H),2.91(br s,2H),2.81(s,2H)。ES(+)MS m/e＝614(M+H)⁺。

i) To a solution of 21.9(70.3mg,0.115mmol) in THF (1.00mL) was added LiOH (1.0M in water, 0.360mL,0.360 mmol). The resulting mixture was stirred at room temperature until TLC and LC/MS showed complete hydrolysis (-0.5 hours). The reaction was then quenched by the addition of 1.0m aqueous cl (0.400mL) and concentrated to dryness. The residue was dissolved in dimethyl sulfoxide ("DMSO"; 4.0mL) and purified by preparative RP-HPLC. Fractions containing pure compound were combined and concentrated. The residue was lyophilized under high vacuum for 48 hours to give 33.8mg (49%) of compound 21 as a white powder.¹H NMR(CDCl₃)7.46-7.39(m,4H),7.28(m,2H),7.15(br s,1H),6.77(s,1H),6.38(s,1H),5.74(br s,2H),5.04(m,1H),4.83(br s,1H),4.53(m,3H),3.69(m,1H),3.00(s,3H),2.85(br s,2H)。ES(+)MS m/e＝600(M+H)⁺。

Example 22

This example describes the synthesis of the following compounds

Prepared according to scheme 14 and following procedures.

Reaction scheme 14

a) 5.0g (28.2mmol) of compound 22.1(Plobeck et al, J.Med.chem). 43:3878-3894(2000)) and sulfuryl chloride (100 mmol each, added at the beginning of the reaction and 15 hours later) were refluxed for 36 hours in acetic acid (50 mL). The off-white solid from the concentrated reaction mixture was rinsed with ether and DCM (50mL) was added to the crude product followed by BBr₃(1.0MIn DCM, 100 mL). After 6 hours, the reaction mixture was concentrated and water (50mL) was added carefully. The resulting precipitate was collected by filtration under reduced pressure and dried to give crude compound 22.2 in quantitative yield.

b) The crude compound 22.2 was dissolved in DCM/pyridine (50mL/50mL) and cooled to 0 ℃. Trifluoromethanesulfonic anhydride (42.3mmol) was added slowly thereto, and then the reaction was stirred at room temperature for 6 hours. The reaction mixture was partitioned between ethyl acetate (200mL) and water (50mL), and the organic layer was washed with water (30mL, twice) and brine, dried over anhydrous sodium sulfate and filtered. The residue obtained by concentrating the filtrate was purified by silica gel column chromatography to give compound 22.3(3.16g, 32%). ESI-MS (M/z): (M + H)⁺)364/366。

c) Carbon monoxide was bubbled through a mixture of compound 22.3(581mg,1.6mmol), BiNAP (0.2mmol), palladium acetate (0.2mmol), triethylamine (1mL), anhydrous methanol (3mL) and anhydrous DMF (3mL) for 10 min, then the reaction was heated under a carbon monoxide balloon at 65 ℃ for 15 h. The reaction mixture was partitioned between ethyl acetate (100mL) and water (25mL), and the organic layer was washed with water (25mL, twice) and brine, dried over anhydrous magnesium sulfate and filtered. The residue obtained by concentrating the filtrate was purified by means of a silica gel column to give compound 22.4(213mg, 49%). ESI-MS (M/z): (M + H)⁺)274/276。

d) To a suspension of sodium hydride (24mg,1.0mmol) in THF (2mL) were added compound 22.4(63mg,0.23mmol), 4-chlorobenzyl chloride (55mg,0.34mmol) and tetrabutylammonium iodide (10 mg). After 6 hours, the reaction was diluted with ether and filtered through silica gel, rinsing with ether. The residue obtained by concentrating the filtrate was purified by means of a silica gel column to give compound 22.5(50mg, 55%). ESI-MS (M/z): (M + H)⁺)398/400。

e) A mixture of compound 22.5(50mg) and 1mmol of LiI in 2mL of pyridine was refluxed overnight. The reaction was concentrated in vacuo and the residue was dried under high vacuum for 2 hours. The crude compound 22.6 obtained was used without further purification. ESI-MS (M/z): (M +1), 384.

f) Compound 22 was prepared according to example 3g, except that compound 22.6 was used instead of compound 3.7 (yield: 82%).¹H NMR(400MHz,dmso-d₆):9.12(d,1H),8.54(t,1H),7.90(s,1H),7.77(dd,1H),7.72(dd,1H),7.42(d,2H),7.35(d,2H),7.16(dd,1H),4.77(m,1H),4.70(s,2H),3.64(m,2H),3.53(t,2H),3.01(t,2H)ppm。ESI-MS(m/z):(M+H⁺)580。

Example 23

This example describes the synthesis of the following compounds

Prepared according to scheme 15 and following procedures.

Reaction scheme 15

a) To a solution of Trit-Ser-Ome (compound 23.1,10mmol) and triethylamine in DCM (40mL) was added methanesulfonyl chloride (11mml) slowly, after 12 h the reaction was extracted with ether (100mL), washed with water, dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated to give compound 23.2, which was used without further purification.

b) A solution of crude compound 23.2, sodium azide (20mmol) in DMF was stirred for 15 h. The reaction was extracted with ether (100mL), washed with water, dried over magnesium sulfate and filtered. The residue obtained by concentrating the filtrate was purified by column chromatography eluting with 0-20% ethyl acetate in hexane to give compound 23.3.

c) 1mmol of compound 23.3, 1.5mmol of cyclopropylacetylene, 0.02mmol of CuI and 0.02mmol of Et₃N in 6mL CH₃The mixture in CN was stirred at rt overnight. The solvent was removed and the residue was purified to give compound 23.4 in 65% yield.¹H NMR(400MHz,CD₃OD):7.80(s,1H),7.29-7.31(m,6H),7.15-7.23(m,9H),4.49(m,2H),3.73(m,1H),3.16(s,3H),1.99(m,1H),0.99(m,2H),0.80(m,2H)ppm；ESI-MS(m/z):(M+H⁺)453.15。

d) A mixture of 0.5mmol of compound 23.4 in 2mL of 4.0N HCl in dioxane was stirred at room temperature for 1 hour. The solvent was removed and the residue was diluted with 10mL of water. The mixture was extracted 3 times with ether and the aqueous phase was dried with a lyophilizer to give compound 23.5 in quantitative yield. ESI-MS (M/z): (M + H)⁺)212.15。

e) Compound 23 is prepared according to example 3g, except that compound 4.4 is used instead of compoundCompound 3.7, compound 23.5 instead of compound 3.4.¹H NMR(400MHz,CD₃OD) 7.74(s,1H),7.33 and 7.49(m,5H),5.25(m,1H),4.63-4.92(m,4H),3.99 and 3.68(m,2H),2.89(m,2H),1.91(m,1H),0.95(m,2H),0.75(m,2H) ppm; ESI-MS (M/z): (M + H)⁺)562.10。

Example 24

This example describes the synthesis of the following compounds

Prepared according to scheme 16 and following procedures.

Reaction scheme 16

a) To a solution of 50mmol of 1-bromo-3, 5-difluorobenzene in 100mL of anhydrous DMF at 0 deg.C was added 50mmol of NaSCH3, and the resulting mixture was stirred overnight at room temperature and with 10mL of saturated NH₄And (4) treating with a Cl aqueous solution. The mixture was diluted with 1L of water and extracted several times with hexane. Washing the extract with water and anhydrous Na₂SO₄And (5) drying. The solvent was removed and the residue was purified to give compound 24.2 in 90% yield.¹H NMR(400MHz,CD₃Cl):7.15(s,1H),7.02(d,J＝8.3Hz,1H),6.89(d,J＝9.2Hz,1H),2.50(s,3H)ppm。

b) A mixture of 40mmol of compound 24.2 and 42mmol of CuCN in 100mL of anhydrous DMF was stirred at 150 ℃ overnight. The mixture was diluted with 500mL of water and extracted several times with ether. The mixture is then diluted with NH₄Aqueous OH and Water washing with anhydrous Na₂SO₄And (5) drying. The solvent was removed and the residue was purified to give compound 24.3 in 50% yield.¹H NMR(400MHz,CD₃Cl):7.28(s,1H),7.17(d,J＝9.2Hz,1H),7.11(d,J＝6.8Hz,1H),2.53(s,3H)ppm；ESI-MS(m/z):(M+H⁺)168.0。

c) 20mmol of compound 24.3 and 22mmol of KOH in 25mL of EtOH and 35mL of H₂The mixture in O was stirred at 60 ℃ for 30 minutes. The mixture was concentrated and the residue diluted with 100mL of water and extracted several times with EtOAc. Anhydrous Na for extract₂SO₄And (5) drying. The solvent was removed and the residue was dried in vacuo to give crude compound 24.4. The crude product was used in the next step without further purification. ESI-MS (M/z): (M + H)⁺)187.0。

d) To a solution of 20mmol of compound 24.4 in 60mL of anhydrous THF at 0 deg.C was added 24mmol of LiAlH₄(1.0M in THF). After stirring overnight at room temperature, saturated NH was carefully added to the reaction mixture₄Aqueous Cl solution. The resulting suspension was then concentrated. The residue was dissolved in 200mL of 1.0N HCl and extracted several times with EtOAc. Extracting with anhydrous Na₂SO₄Dried and concentrated. The residue was then purified to give 5-fluoro-3-methylsulfanyl-1-benzyl alcohol in 81% yield.¹H NMR(400MHz,CD₃Cl):7.03(s,1H),6.86(m,2H),4.69(s,2H),2.51(s,3H)ppm；ESI-MS(m/z):(M+H⁺)173.1。

15mmol of 5-fluoro-3-methylmercapto-1-benzyl alcohol and 20mmol of SOCl₂In 30mL of anhydrous CH₂Cl₂The mixture in (1) is refluxed for several hours. The mixture was then diluted with 100mL of CH₂Cl₂Diluting with saturated NaHCO₃Aqueous solution, saturated NH₄Washed with aqueous Cl solution, brine and anhydrous Na₂SO₄And (5) drying. The solvent was removed and the residue was purified to give compound 24.5 in 85% yield.

e) Compound 24.6 was prepared according to examples 3a-c, except that compound 24.5 was used instead of 3-methylthiobenzyl chloride.

f) Compound 24 was prepared according to example 3g, except that compound 24.6 was used instead of compound 3.7.¹HNMR(400MHz,CD₃OD)7.92(s,1H), 7.80(s,1H),7.75(d, J ═ 7.83Hz,1H),7.74(s,1H),7.59(d, J ═ 7.34Hz,1H),7.49(d, J ═ 9.29Hz,1H),7.07 and 7.35(m,2H),6.96(s,1H),5.10(dd, J ═ 9.78,4.40Hz,1H),4.72 and 4.91(m,2H),3.77 and 4.00(m,2H),3.50(dd, J ═ 14.18,4.40Hz,1H),3.20(m,1H),2.91(m,2H) ppm; ESI-MS (M/z): (M + H)⁺)633.10。

Example 25

This example describes the synthesis of the following compounds

It is prepared according to scheme 17 and the following procedure.

Reaction scheme 17

a) 5.0mmol of Compound 3.2 (example 3a), 0.25mmol of Pd (PPh) at 0 deg.C₂)₂Cl₂0.25mmol of CuI in 15mmol of degassed Et₃N and 40mL of degassed toluene 5.5mmol of isobutyryl chloride was added. The resulting mixture was stirred at room temperature overnight and with 20mL of saturated NaHCO₃And (4) treating with an aqueous solution. The organic layer was separated and dried over anhydrous Na₂SO₄And (5) drying. The solvent was removed and the residue was purified by column chromatography to give compound 25.1, 80% yield.¹H NMR(400MHz,CD₃OD):4.38(m,1H),3.74(s,3H),2.91(m,2H),2.61(m,1H),1.44(s,9H),1.14(d,J＝6.85Hz,6H)ppm；ESI-MS(m/z):(M+H⁺)320.01。

b) To 1.0mmol of Compound 25.1 at 0 deg.C in 3mL of CH₃Adding 1.0mmol of NH into the solution in OH₂NH₂The resulting mixture was stirred for an additional 30 minutes. Removing the solvent and purifying the residueCompound 25.2 was obtained in 65% yield.

¹H NMR(400MHz,CD₃OD) 6.28 and 5.95(s, s,1H),4.39 and 4.19(m,1H),3.72(s,3H),2.91(m,2H),2.70 and 2.98(m,1H),1.43 and 1.45(s, s,9H),1.12 and 1.27(m,6H) ppm; ESI-MS (M/z): (M + H)⁺)312.20。

c) A mixture of 0.5mmol of compound 25.2 in 4mL of 4.0N HCl in dioxane was stirred at room temperature for 12 hours. The solvent was removed and the residue was dried in vacuo to give compound 25.3. ESI-MS (M/z): (M + H)⁺)212.10。

d) Compound 25 was prepared according to example 3g, except that compound 4.4 was used instead of compound 3.7 and compound 25.3 was used instead of compound 3.4.

¹H NMR(400MHz,CD₃OD) 7.15-7.54(m,5H),6.38(s,1H),5.06(dd, J ═ 9.78,4.89Hz,1H),4.66 and 4.88(m,2H),4.01 and 3.71(m,2H),3.41(dd, J ═ 15.41,4.65Hz,1H),3.18(dd, J ═ 15.16,4.65Hz,1H),3.07(m,1H),2.92(m,2H),1.32(d, J ═ 7.34Hz,6H) ppm; ESI-MS (M/z): (M + H)⁺)563.10。

Example 26

This example describes the synthesis of the following compounds

It is synthesized according to scheme 18 and the following procedure.

Reaction scheme 18

a) Reaction of 26.1(12.4g,75mmol) with NH₄BF₄(10.5g,100mmol) in water (85mL) with concentrationTreated with HCl (15mL), cooled to 3 ℃ and treated by dropwise addition of NaNO over 25 minutes₂(5.18g,75mmol) in water (12 mL). The resulting thick slurry was stirred for 35 minutes, the solid was collected by filtration, rinsed with water, methanol and ether, and concentrated under N₂And (4) drying. The solid was added in one portion to a stirred mixture of KOAc (8.1g,82.5mmol) and 18-crown-6 (0.5g,1.9mmol) in chloroform (170 mL). After 70 minutes, water (170mL) was added and the layers were separated. The aqueous phase was extracted with chloroform and the combined organic layers were rinsed with water, dried and concentrated. The residue was triturated with hexane and the resulting solid was isolated by filtration to give 8.85g (67% yield) of compound 26.2 as a dark yellow powder.¹H NMR(CDCl₃)3.96(s,3H),7.80–7.85(m,2H),8.14(s,1H),8.27(s,1H)；ES(+)MS m/e＝177(M+1)。

b) A solution of compound 26.2(5.0g,28.4mmol) in THF (56mL) was treated with LiOH (21mL of a 2M aqueous solution, 42mmol) and the reaction mixture was stirred at 50 ℃. After 4 hours, the reaction mixture was cooled to room temperature and diluted with water. The alkaline aqueous layer was rinsed with ether, adjusted to pH 3-4 by addition of 1M HCl, and extracted with ethyl acetate. The aqueous layer was extracted with ethyl acetate and the combined organic layers were rinsed with brine and MgSO₄Drying and concentration gave 4.0g (87% yield) of compound 26.3.¹H NMR(CD₃OD)7.79–7.87(m,2H),8.14(s,1H),8.29(s,1H)；ES(+)MS m/e＝163(M+1)。

c) A solution of compound 1.1(7.5g,20.8mmol) in DCM (30mL) was treated with TFA (10 mL). After 1 hour, the reaction mixture was concentrated to give 7.8g (100% yield) of compound 26.4. ES (+) MS M/e 261(M + 1).

d) A solution of compound 26.3(7.8g,20.8mmol), compound 26.4(3.4g,20.8mmol), 1-hydroxybenzotriazole hydrate ("HOBt",3.5g,22.3mmol), diisopropylethylamine ("DIEA",14mL,83.3mmol) in DMF (100mL) was treated with EDCI (4.4g,22.3 mmol). After 2 hours, the reaction mixture was treated with 1M HCl and extracted with ethyl acetate. The combined organic extracts were extracted with NaHCO₃(saturated) rinse, rinse with brine, rinse with water, MgSO₄Drying and concentrating to obtain 8.4g (99% yield) of the title compound. ES (+) MS M/e 404(M + 1).

e) A solution of compound 26.5(8.4g,20.8mmol) in pyridine (70mL) was treated with lithium iodide (11.1g,83.1mmol) and the reaction mixture was heated to 100 ℃. After 16 hours, the reaction mixture was cooled to room temperature and diluted with 1M NaOH (aq). The basic aqueous layer was rinsed with ether to remove most of the pyridine. The aqueous portion was then carefully acidified with concentrated HCl to pH 3-4. The resulting slurry was filtered. The precipitate was collected and dissolved in THF, and the filtrate was extracted with ethyl acetate. The THF and ethyl acetate solutions were combined, rinsed with brine, MgSO₄Drying and concentration gave 7.1g (88% yield) of compound 26.6. ES (+) MS M/e 390(M + 1).

f) A solution of compound 26.6(3.06g,7.83mmol) and DIEA (4.6mL,25.4mmol) in dimethylformamide ("DMF") was treated with HATU (3.06g,8.06mmol) and the resulting mixture was stirred at room temperature. After 20 minutes, the reaction mixture was treated sequentially with HCl. H-DAP (Boc) -OMe (2.18g,8.59mmol) and N, N-dimethylaminopyridine ("DMAP",0.568g,4.65 mmol). After 2.5 hours, the reaction was diluted with ethyl acetate, washed three times with water, once with brine, MgSO₄Drying and concentrating. Flash column chromatography gave 3.91g (84% yield) of compound 26.7.¹H NMR (400MHz, chloroform-d)1.42(s,9H),2.81(s,2H),3.70(m,2H),3.75(2H),3.81(s,3H),4.82(m,21H),4.99(m,1H),7.21(d,2H),7.59(s,1H),7.81(d,1H),8.10(s,1H)。MS(API-ES⁺)m/z:590.2(M+H⁺) 534.1(M-t butyl + H)⁺),490.1(M-Boc+H⁺)。

g) A solution of compound 26.7(3.91g,6.62mmol) in DCM was treated with HCl (8.3mL of 4M dioxane, 33.2mmol) and the resulting mixture was stirred at RT. After 2 hours, the reaction mixture was concentrated to give the HCl salt, which was used without further purification. The HCl salt (3.94g,6.99mmol) and triethylamine ("TEA",3.0mL,21.5mmol) in methanol were treated with N-cyanoimido-S, S-dimethyl-dithiocarbonic acidEster (1.37g,8.43mmol) and the reaction mixture was stirred at 50 ℃. After 3.5 hours, the reaction mixture was concentrated to remove most of the methanol, diluted with ethyl acetate, washed twice with water, once with brine, MgSO₄Drying and concentrating. Flash column chromatography gave 3.27g (80% yield) of Compound 26.8(MS (API-ES)⁺)m/z:588.2(M+H⁺). A solution of compound 26.8(0.10mmol) in 4:1 methanol/dichloroethane ("DCE",2.5mL) was prepared and treated sequentially with 2M methanol-ammonia (0.25mmol) and silver nitrate (0.10 mmol). The reaction mixture was stirred at 50 ℃ until the conversion was complete by LCMS. The reaction mixture was then filtered through celite and KOH (0.1mL of 2M methanol solution, 0.2mmol) was added. The reaction mixture was stirred again at 50 ℃. After 2-4 hours, the reaction mixture was directly subjected to preparative HPLC purification to afford compound 26.

Example 27

This example describes the synthesis of the following compounds

Wherein R is^AAnd R^BEach independently hydrogen, an aliphatic moiety, an aromatic moiety, a heteroaromatic moiety, or together form a cyclic moiety. These compounds are prepared according to the method of example 26, except that in step g the formula HNR is used^AR^BSubstituted amines of (a) in place of ammonia. Illustrative examples of substituted amines and resulting compounds are shown in table 1.

TABLE 1

Example 28

This example describes the synthesis of the following compounds

Prepared according to scheme 19 and following procedures.

Reaction scheme 19

a) As indicated above, according to Okada, t. et al, chem.phar.ball., 1993,41(1), 126-; the method disclosed by Frigola, j, et al, j.med.chem., 1993,36(7), 801-. A solution of commercially available 28.1 in DCM in the presence of a base was treated with trifluoromethanesulfonyl chloride to give 36.2. The product was then dissolved in dimethoxyethane ("DME") with dimethylamine replacing the mesylate moiety. Finally, in MeOH at 45PSI H₂(g) Pd/C-catalyzed hydrogenation gave compound 28.4.

b) Compound 28 was synthesized according to the procedure for example 26, except that 3- (dimethylamino) cyclobutanol (35.1) was used in place of ammonia in step g.

Example 29

This example describes the synthesis of the following compounds

It is prepared according to scheme 20 and the following procedure.

Reaction scheme 21

a) To a solution of benzyl 3-pyrroline-1-carboxylate (compound 29.1,10mmol) in THF (15mL) was added N-methylmorpholine (22mmol) and O_SO₄(2mL, 2.5 wt% in t-BuOH), the resulting mixture was stirred at room temperature overnight. The solvent was removed and the residue was dissolved in EtOAc (100mL) and diluted Na₂SO₃Aqueous solution, saturated NH₄Washing with aqueous Cl solution and brine, and washing with anhydrous Na₂SO₄And (5) drying. The solvent was removed and the residue was purified by column chromatography to give compound 29.2, 55% yield. EIMS (m/z): C₁₂H₁₅NO₄(M⁺) + Na calculated 260.1, found 260.1;¹H NMR(CD₃OD,400MHz):7.31-7.38(m,5H),5.13(s,2H),4.17(m,2H),3.58(m,2H),3.34(m,2H)ppm。

b) a mixture of compound 29.2(1.0mmol) and 10% Pd/C (0.1mmol) in methanol (5mL) was taken up in H at room temperature₂Stirred under atmosphere for several hours. The reaction mixture was filtered, the filtrate was concentrated and the residue was dried in vacuo to give compound 29.3.

c) Compound 29 was synthesized according to the procedure for example 26, except that (3R,4S) - (dihydroxy) pyrrolidine (29.3) was used in place of ammonia in step g.

Example 30

This example describes the synthesis of the following compounds

It is prepared according to scheme 21 and the following procedure.

Reaction scheme 21

a) Mixing (3R,4R) -benzyl-3, 4-pyrrolidine diol (compound 30.1,1mmol) and 20% Pd (OH)₂A mixture of/C (0.1mmol) in methanol (10mL) at room temperature at 45psi of H₂(g) Shake down for several hours. The reaction mixture was filtered, the filtrate was concentrated and the residue was dried in vacuo to give compound 30.2.

b) Compound 30 was synthesized according to the procedure for example 26, except that compound 30.2 was used in place of ammonia in step g.

Example 31

This example describes the synthesis of the following compounds

It is prepared according to scheme 22 and the following procedure.

Reaction scheme 22

a) A mixture of commercially available benzyl 3-pyrroline-1-carboxylate (compound 28.1,10mmol) and m-CPBA (12mmol) in DCM (50mL) was stirred at room temperature overnight. The reaction mixture was diluted with DCM (100mL) and successively saturated Na₂SO₃Aqueous solution, brine washing. Anhydrous Na for organic layer₂SO₄Dried and then concentrated. The residue was purified by chromatography to give compound 31.1, 80% yield. EIMS (m/z): C₁₂H₁₃NO₃(M⁺) + Na calcd 242.1, found 242.1;¹H NMR(CDCl₃,400MHz):7.36-7.37(m,5H),5.13(s,2H),3.89(m,2H),3.70(m,2H),3.41(m,2H)ppm。

b) compound 31.1(5mmol) in concentrated NH₃The mixture in solution (20mL) was stirred at 65 ℃ overnight. The reaction mixture was concentrated and dried in vacuo to afford compound 31.2. This material was used without further purification.

c) Compound 31.2(10mmol) and Et at-20 deg.C₃A solution of N (20mmol) in dry THF (100mL) was treated by dropwise addition of TFAA (10mmol) over 1 hour. After 1 hour, the reaction mixture was saturated with NH₄Aqueous Cl (1 mL). The solvent was removed and the residue was dissolved in DCM (100 mL). The mixture was then successively saturated with NH₄Aqueous Cl solution, saturated NaHCO₃Aqueous solution, brine washing. Anhydrous Na for organic layer₂SO₄Drying, removal of the solvent and purification of the residue by chromatography gave compound 31.3.

d) Compound 31.3(5mmol) and Et at 0 deg.C₃A solution of N (10mmol) in dry DCM (20mL) was treated by dropwise addition of MsCl (5.5mmol) and the compound was gradually warmed to room temperature. After 1 hour at room temperature, the reaction mixture containing in situ generated 31.4 was treated with DBU (30mmol) and the resulting mixture was stirred for several hours. The solvent was removed and the residue was purified by chromatography to give compound 31.5.

e) The reaction of compound 31.5(3mmol) and K₂CO₃(6mmol) MeOH/H at 2/1(v/v)₂The mixture in O (15mL) was stirred at room temperature. After 24 hours, the solvent was removed and the residue was taken up with saturated NaHCO₃Aqueous solution (20mL) and the mixture was extracted several times with DCM. Extracting with Na₂CO₃Drying, removal of the solvent and purification of the residue by chromatography gave compound 31.6.

f) The compound 31.6(2mmol), NaHCO₃(3mmol)、Boc₂A mixture of O (2.2mmol) in 1:1, 4-dioxane/water (20mL) was stirred at room temperatureFor several hours. The mixture was diluted with brine (50mL) and extracted several times with EtOAc. The combined extracts were washed with brine and anhydrous Na₂SO₄Drying and concentrating. The residue was purified by chromatography to afford the N-Boc protected intermediate. A mixture of this intermediate (1mmol) and 10% Pd/C (0.1mmol) in MeOH (5mL) at room temperature in H₂(g) Stirred under atmosphere for several hours. The reaction mixture was filtered and the filtrate was concentrated and the residue was dried in vacuo to give compound 31.7.

g) Compound 31 was prepared according to example 26g, except that ammonia was replaced with compound 31.7.

Example 32

This example describes the synthesis of the following compounds

Prepared according to scheme 23 and following procedures.

Reaction scheme 23

a) The compound 31.2(2mmol), NaHCO₃(3mmol)、Boc₂A mixture of O (2.2mmol) in 1:1, 4-dioxane/water (20mL) was stirred at room temperature for several hours. The mixture was diluted with brine (50mL) and extracted several times with EtOAc. The combined extracts were washed with brine and anhydrous Na₂SO₄Drying and concentrating. The residue was purified by chromatography to give compound 32.3.

b) A mixture of compound 32.3(1mmol) and 10% Pd/C (0.1mmol) in MeOH (5mL) at room temperature in H₂(g) Stirred under atmosphere for several hours. The reaction mixture was filtered, the filtrate was concentrated, and the residue wasAir-dried to give compound 32.4.

c) Compound 32 is prepared according to example 26g, except that ammonia is replaced by compound 32.4.

Example 33

This example describes the synthesis of the following compounds

Prepared according to scheme 24 and following procedures.

Reaction scheme 24

a) A solution of compound 33.1(10mmol) and DIEA (25mmol) in DCM (20mL) was treated by dropwise addition of benzyl chloroformate (10mmol) at 0 deg.C, and the reaction mixture was allowed to warm to room temperature. After 2 hours at room temperature, the reaction mixture was diluted with ethyl acetate (100mL), rinsed with 1M HCl (50mL), rinsed with brine, MgSO₄Drying and concentration gave compound 33.2.

b) A solution of compound 33.2(10mmol) and N-methylmorpholine (22mmol) in THF (15mL) was treated with O_SO₄(2mL, 2.5 wt% in t-BuOH) and the resulting mixture was stirred at room temperature overnight. The solvent was removed. The residue was dissolved in EtOAc (100mL) and diluted Na₂SO₃Aqueous solution, saturated NH₄Washing with aqueous Cl solution, brine, and then anhydrous Na₂SO₄And (5) drying. The solvent was removed and the residue was purified by column chromatography to give the title compound.

c) A mixture of compound 33.3(1.0mmol) and 10% Pd/C (0.1mmol) in methanol (5mL) was taken up in H at room temperature₂Stirred under atmosphere for several hours. The reaction mixture was filtered, the filtrate was concentrated and the residue was dried in vacuo to give compound 33.4.

d) Compound 33 was prepared according to example 26g, except that ammonia was replaced with compound 33.4.

Example 34

This example describes the synthesis of the following compounds

It is prepared according to scheme 25 and the following procedure.

Reaction scheme 25

A mixture of Tl (OAc) (17.6g,54.5mmol) in anhydrous acetic acid (40mL) was stirred at reflux for 1 hour and then cooled to room temperature. Compound 32.2(34.6mmol) and iodine (8.46g,33.3mmol) were added and the resulting suspension was heated to reflux. After 9 hours, the reaction mixture was cooled to room temperature. The TlI precipitate was removed by filtration and rinsed with ether. The filtrate was concentrated and the residue was dissolved in ethyl acetate over MgSO₄Drying and concentration gave compound 34.1.

b) A mixture of 34.1(1.0mmol) and 10% Pd/C (0.1mmol) in methanol (5mL) was taken up in H at room temperature₂Stirred under atmosphere for several hours. The reaction mixture was filtered, the filtrate was concentrated and the residue was dried in vacuo to give compound 34.2.

c) Compound 34 was prepared according to example 26g, except that ammonia was replaced with compound 34.2.

Example 35

This example describes the synthesis of the following compounds

It is prepared according to scheme 26 and the following procedure.

Reaction scheme 26

a) A mixture of compound 35.1(10mmol) in ethanol (20mL) was saturated with HCl (g) and stirred at room temperature overnight. The solvent was removed, the residue dissolved in DCM (100mL), the resulting solution treated sequentially with TEA (30mmol) and BnBr (11mmol), and the reaction mixture heated to reflux. After 12 hours, the reaction mixture was cooled to room temperature and concentrated. The residue was dissolved in EtOAc (150mL), washed with brine, anhydrous Na₂SO₄And (5) drying. The solvent was removed and the residue was purified by chromatography to give compound 35.2.

b) Reacting LiBH₄A solution of (20mmol) in dry THF (20mL) was treated dropwise at room temperature with a solution of compound 35.2(5mmol) in THF (5 mL). After stirring overnight, the reaction mixture was quenched by the addition of a few drops of water. The mixture was concentrated, diluted with brine (50mL), and extracted several times with 9/1(v/v) EtOAc/i-PrOH. Mixing the extractive solutions with anhydrous Na₂SO₄Drying, removal of the solvent and purification of the residue by chromatography gave compound 35.3.

c) A solution of compound 35.3(2mmol) and TEA (2.2mmol) in dry THF (10mL) at-78 deg.C was treated by the dropwise addition of trifluoroacetic anhydride ("TFAA",2.2 mmol). After a few hours, the mixture was treated with TEA (6mmol) and the reaction mixture was heated to reflux. The mixture was then concentrated, the residue dissolved in THF (10mL) and washed with water(2.5 mL). The mixture was treated with NaOH (10mmol) at room temperature under vigorous stirring for several hours. The solvent was removed and the residue was taken up with saturated NaHCO₃(20mL) treated with aqueous solution and extracted several times with 9/1(v/v) EtOAc/i-PrOH. Mixing the extractive solutions with anhydrous Na₂SO₄Drying, removal of the solvent and purification of the residue by chromatography gave compound 35.4.

d) Reaction of 35.4(1mmol) with 20% Pd (OH)₂A mixture of/C (0.1mmol) in methanol (10mL) at room temperature at 45psi of H₂(g) Shaking for several hours. The reaction mixture was filtered, the filtrate was concentrated and the residue was dried in vacuo to give compound 35.5.

e) Compound 35 was prepared according to example 26g, except that ammonia was replaced with compound 35.5.

Example 36

This example describes the synthesis of the following compounds

It was prepared according to example 35, except that (2R,4S) -4-hydroxyproline (trans-D-Hyp-OH) was used instead of compound 35.1.

Example 37

This example describes the synthesis of the following compounds

It was prepared according to example 35, except that (2S,4R) -4-hydroxyproline (trans-L-Hyp-OH) was used instead of compound 35.1.

Example 38

This example describes the synthesis of the following compounds

It is prepared according to scheme 27 and the following procedure.

Reaction scheme 27

a) A solution of HCl. H-DAP (Boc) -OMe (10mmol) and DIEA (11mmol) in DCM (50mL) was treated with dropwise addition of benzoyl chloride (11mmol) at 0 ℃. The reaction mixture was allowed to warm to room temperature over 3 hours. The reaction mixture was concentrated and the residue was purified by chromatography. The intermediate was treated with 4M HCl in dioxane and the resulting mixture was stirred at room temperature. After 1 hour, the solvent was removed to give compound 38.1, which was used without further purification.

b) A solution of compound 38.1(10mmol) and DIEA (12mmol) in DCM (50mL) was treated with p-nitrophenyl chloroformate (11mmol) dropwise at 0 deg.C, and the reaction mixture was allowed to warm to room temperature. After 2 hours, the reaction mixture was concentrated and the residue was purified by chromatography to give compound 38.2.

c) A solution of compound 38.2(10mmol) and TEA (25mmol) in 1:1 DCE/DMF (10mL) was treated with 2M methanolic-aqueous ammonia (18mmol) and the mixture was heated to 40 ℃. After 15 hours, the reaction mixture was concentrated and the residue was purified by chromatography to give compound 38.3.

d) A mixture of compound 38.3(1.0mmol) and 10% Pd/C (0.1mmol) in methanol (10mL) was treated with H at 45psi at room temperature₂(g) Shaking down for several hours. The reaction mixture was filtered, the filtrate was concentrated and the residue was dried in vacuo to give compound 38.4.

e) A solution of compound 26.6(8.0mmol) and DIEA (25mmol) in DMF was treated with HATU (8.0mmol) and the resulting mixture was stirred at room temperature. After 20 minutes, the reaction mixture was treated sequentially with compound 38.4(8.6mmol) and DMAP (0.5mmol) and the mixture was then heated to 60 ℃. After 2.5 hours, the reaction was diluted with ethyl acetate, washed three times with water, once with brine, MgSO₄Drying and concentrating. Flash column chromatography gave compound 38.5.

f) A solution of compound 38.5(0.15mmol) in methanol (1mL) was treated with 2M methanol-KOH (0.45mmol) and the reaction mixture was heated to 50 ℃. After 3 hours, the reaction mixture was concentrated to dryness and the residue was purified by preparative HPLC to give compound 38.

Example 39

This example describes the synthesis of the following compounds

Wherein R is^AAnd R^BEach independently hydrogen, an aliphatic moiety, an aromatic moiety, a heteroaromatic moiety, or together form a cyclic moiety. These compounds are prepared according to the method of example 38, except that in step c the compound of formula HNR is used^AR^BSubstituted amines of (a) in place of ammonia. Illustrative examples of substituted amines and resulting compounds are shown in table 2.

TABLE 2

Example 40

This example describes the synthesis of the following compounds

It is prepared according to scheme 28 and the following procedure.

Reaction scheme 28

a) Commercial compound 40.1(10mmol) in THF (50mL) was treated with sodium bisulfite (50mmol) in water (20 mL). After 8 hours of reaction at room temperature, the reaction was extracted with ethyl acetate (100mL), and the organic extract was washed with water and brine, dried over anhydrous magnesium sulfate and filtered to give crude compound 40.2.

b) To a slurry of compound 40.2(10mmol), ammonium tetrafluoroborate (12.5mmol) in water (12mL) was added concentrated HCl (2 mL). The reaction was cooled to 0 ℃ C, and sodium nitrite (10mmol) was added thereto. After the reaction was stirred at 0 ℃ for 1 hour, the solid was collected by filtration, rinsed with methanol, ether and dried under vacuum. The resulting solid was added to a stirred solution of HOAc (3mL), 18-crown-6 (0.3mmol) in chloroform (20 mL). After 1 hour, water (10mL) and DCM (20mL) were added. The organic layer was separated, dried over magnesium sulfate and filtered. The filtrate was concentrated and the resulting residue triturated with hexanes to give product 40.3.

c) Compound 40.4 was prepared according to the procedure used to prepare compound 6.1, except that R-3- (+) -pyrrolidinol was used instead of pyrrolidine.

d) Compound 40.4 was dissolved in DCM and treated with triethylamine (1.5eq) and acetic anhydride (1.2 eq). The resulting solution was filtered through silica gel and concentrated. The residue was then purified by silica gel column chromatography to give compound 40.5.

e) Compound 40.5 in DCM was treated with anhydrous 4N HCl in dioxane (2.0 eq). After disappearance of the starting material, the reaction was concentrated to give compound 40.6.

f) Compound 40.7 was prepared according to the procedure used to prepare compound 1.5, except that compound 40.6 was used instead of 1.3.

g) Compound 40.8 was prepared according to the procedure used to prepare compound 1.6, except that compounds 40.7 and 40.3 were used instead of compounds 1.5 and 1.11.

h) Compound 40 was prepared according to the procedure used to prepare compound 1, except that compound 40.8 was used instead of 1.6.

EXAMPLE 41

This example describes the synthesis of the following compounds

It is prepared according to scheme 29 and the following procedures.

Reaction scheme 29

Compound 41 was prepared according to the procedure used for the preparation of compound 3, except that compounds 18.10 and 40.6 were used instead of 3.4 and 3.7. The enantiomerically pure compounds were separated using chiral column chromatography.

Example 42

This example describes the synthesis of the following compounds

It is prepared according to scheme 30 and the following procedure.

Reaction scheme 30

a) A mixture of 18.7(10mmol), 1-ethoxy-1-vinyltributyltin (10.5mmol), Pd (PPh)₄A solution (0.5mmol) in dimethoxyethane (DME,50mL) was heated to 80 deg.C until compound 18.7 disappeared. The reaction was cooled to room temperature, and 4N aqueous HCl (5mL) was added thereto. The reaction was stirred for 3 hours and extracted with ether (80 mL). The organic extract was washed with brine, dried over anhydrous magnesium sulfate, filtered and concentrated. The residue was purified by column chromatography to give compound 42.1.

b) A solution of compound 42.1 in THF at-78 deg.C was treated with LDA (2.0 eq). After 1 hour, a solution of compound 18.2(1.0eq) in THF was added to the dry ice cooled reaction. After a further 3 hours, saturated NH was added to the reaction₄Aqueous Cl and the mixture was allowed to warm to room temperature. The reaction mixture was partitioned between ethyl acetate and water, and the organic layer was washed with water and brine, dried over anhydrous magnesium sulfate and filtered. The residue obtained by concentrating the filtrate was purified by silica gel column to obtain compound 42.2.

c) Compound 42.2 in ethanol was treated with sodium borohydride (2.0 eq). After 1 hour, the reaction mixture was partitioned between ethyl acetate and water, and the organic layer was washed with brine, dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated, and the obtained residue was purified by silica gel column to obtain compound 42.3.

d)42.3 LiI (3eq) in pyridine at reflux overnight. The solvent was removed and the residue was dissolved in EtOAc. The resulting solution was then saturated with NH₄Washing with aqueous Cl solution and anhydrous Na₂SO₄And (5) drying. The solvent was removed and the residue was dried in vacuo to give compound 42.4 in quantitative yield. The crude product was carried on to the next step without further purification.

e) Compound 42 was prepared according to the procedure used to prepare compound 3, except that compounds 42.4 and 40.6 were used instead of 3.4 and 3.7. The enantiomerically pure compounds were separated using chiral column chromatography.

Example 43

This example describes the synthesis of the following compounds

It is prepared according to scheme 31 and the following procedure.

Reaction scheme 31

a) Compound 18.2 in ethanol was treated with hydroxylamine (1.05 eq). After 10 hours, the reaction was concentrated and the residue was dried under vacuum to give compound 43.1.

b) 43.1 catalyst 20% Pd (OH) under 45psi of hydrogen₂hydrogenation/C gives compound 43.2.

c) Commercial 43.3 in carbon tetrachloride was treated with N-chlorosuccinimide (NCS,3 eq). The reaction mixture was then diluted with ethyl acetate, washed with 1N NaOH, water and brine, and then anhydrous MgSO₄Dried and filtered. CoarseThe product was recrystallized from hot ethanol to give compound 43.4.

d) 43.4 in THF was treated with LiOH (2eq,2.0N in water). After most of the starting material was consumed, the mixture was diluted with ethyl acetate, washed with saturated aqueous ammonium chloride solution, water and brine, and then anhydrous MgSO₄Dried and filtered. The crude product was recrystallized from hot ethanol to yield compound 43.5.

e) A solution of 43.5,43.2(1.1eq) and EDC (1.0eq) in DMF was stirred at room temperature for 10 hours. The mixture was then diluted with ethyl acetate, washed with saturated ammonium chloride, water and brine, and then anhydrous MgSO₄Dried and filtered. The crude product was then refluxed with formic acid until LC-MS indicated that the reaction was complete. The solvent was evaporated and the crude product was purified by silica gel column to give compound 43.6.

f) A mixture of 43.6, LiI (3eq) in pyridine was refluxed overnight. The solvent was removed and the residue was dissolved in EtOAc. The resulting solution was then saturated with NH₄Aqueous Cl solution and anhydrous Na₂SO₄And (5) drying. The solvent was removed and the residue was dried in vacuo to give compound 43.7 in quantitative yield. The crude product was carried on to the next step without further purification.

g) Compound 43 was prepared according to the procedure used to prepare compound 3, except that compounds 43.7 and 40.6 replaced 3.4 and 3.7.

Variations in

It will also be appreciated that each of the components used in the synthesis of the compounds of the invention may be varied prior to synthesis or after the construction of the core structure of formula (I). As used herein, the term "altering" or "change" refers to reacting a compound (I) of the invention or any precursor fragment (or any class or subclass thereof) at one or more active sites to modify a functional group or add a functional group (e.g., nucleophilic addition of a substrate). Various reaction schemes are generally described herein for assisting the reader in the synthesis of various compounds through variations in the components of intermediates or through variations in the core structures, classes and subclasses thereof described herein. It is understood that various modifications of the reaction may be employed to produce compounds other than those described in the examples herein. As just a few examples, where a double bond is present in the compound structure, epoxidation and aziridination may be carried out to produce epoxide and aziridine derivatives of the compounds described herein. For additional guidance in the art, practitioners refer to "Advanced Organic Chemistry", March, j., John Wiley & Sons,2001, fifth edition, the entire contents of which are incorporated herein by reference.

2) Biological data:

as noted above, LFA-ICAM interactions may be directly implicated in a variety of inflammatory disease conditions, including but not limited to transplant rejection, dermatitis, psoriasis, asthma, and rheumatoid arthritis. Thus, compounds that modulate the interaction between intracellular adhesion molecules (e.g., ICAM-1, -2, and-3) and the leukocyte integrin family of receptors are useful in the development of novel therapeutic agents. Described below are certain assays for determining ICAM-1: LFA receptor binding, human T cell adhesion, T cell proliferation described in published PCT applications WO 99/49856 and WO 02/05114, the entire contents of which are incorporated herein by reference. WO 99/49856 also describes the preparation and purification of full-length LFA-1 from 293 cells, the preparation of plasmids for expressing human ICAM-1 immunoadhesion (immunoadhesion), and the generation of 293 cell lines expressing ICAM-1 immunoadhesion.

ICAM-1: LFA receptor binding assay (protein/protein assay):

competitive inhibition of the CD11a/CD18-ICAM-1 interaction was determined by adding known amounts of inhibitor to a two protein/protein assay system as described below.

Forward Format LFA-1: ICAM-1 test (PPFF):

purified full-length recombinant human LFA-1 protein was purified in 0.02M Hepes, 0.15M NaCl, 1mM MnCl₂Diluted to 2.5. mu.g/ml and 96-well plates (50. mu.l/well) were plated overnight at 4 ℃. Plates were washed with wash buffer (0.05% Tween in PBS) and incubated at room temperature with 0.02M Hepes, 0.15M NaCl, 1mMMnCl₂Blocking with 1% BSA in (1%). The plate was washed. Add assay buffer (in 0.02M Hepes, 0.15M NaCl, 1mM MnCl₂0.5% BSA) to 2-fold final concentration of inhibitor and incubation at room temperature for 1 hour. 50 μ l/well of purified recombinant human 5 domain ICAM-Ig diluted to 50ng/ml in assay buffer was added and incubated at room temperature for 2 hours. Plates were washed and bound ICAM-Ig was detected using goat anti-huigg (fc) -HRP for 1 hour at room temperature. The plates were washed and developed with 100. mu.l/well TMB substrate for 10-30 min at room temperature. Using 100. mu.l/well of 1M H₂PO₄The colorimetric development was terminated and the plate was read on a plate reader at 450 nM.

The following alternative protein/protein assay system also quantifies competitive inhibition of the CD11a/CD18-ICAM-1 interaction.

PLM2 antibody Capture LFA-1: ICAM-1 assay (PLM2)

Non-functional blocking monoclonal antibody PLM-2 against human CD18 (e.g., Hildreth et al, molecular immunology, Vol.26, No.9, pp.883-895,1989) was diluted to 5. mu.g/ml in PBS and flat bottom 96-well plates were plated with 100. mu.l/well overnight at 4 ℃. Plates were incubated in assay buffer (0.02M Hepes, 0.15M NaCl, 1mM mNCl) at room temperature₂) Blocking with 0.5% BSA in (1) for 1 hour. The plates were washed with 50mM Tris pH 7.5, 0.1M NaCl, 0.05% Tween 20 and 1mM MnCl₂And (6) washing. The purified full length recombinant human LFA-1 protein was diluted to 2. mu.g/ml in assay buffer and 100 ml/well added to the plate and incubated at 37 ℃ for 1 hour. The plate was washed 3 times. Add 50. mu.l/well of inhibitor diluted appropriately in assay buffer to 2-fold final concentration and incubate for 30 min at 37 ℃. Purified recombinant human 5 domain ICAM-Ig (final concentration 80ng/ml) diluted to 161ng/ml in assay buffer at 50 ml/well was added and incubated at 37 ℃ for 2 hours. Plates were washed and bound ICAM-Ig was detected using goat anti-huigg (fc) -HRP for 1 hour at room temperature. The plates were washed and developed with 100. mu.l/well TMB substrate for 5-10 min at room temperature. Using 100. mu.l/well of 1M H₃PO₄The colorimetric development was terminated and the plate was read on a plate reader at 450 nM.

Human T cell adhesion assay (cell attachment assay)

T cell adhesion assays were performed using the human T-lymphoid cell line HuT 78. Goat anti-HuIgG (Fc) was diluted to 2. mu.g/ml in PBS and 96-well plates were coated with 50 ml/well for 1 hour at 37 ℃. Plates were washed with PBS and blocked with 1% BSA in PBS for 1 hour at room temperature. 5 domain ICAM-Ig was diluted to 100ng/ml in PBS and 50. mu.l/well was added to the plate O/N at 4 ℃. HuT78 cells were centrifuged at 100g and the cell pellet was pelleted at 5% CO₂Was treated with 5mM EDTA at 37 ℃ for about 5 minutes. Cells were plated at 0.14M NaCl, 0.02M Hepes, 0.2% glucose and 0.1mM MnCl₂(assay buffer) and centrifuged. Resuspend cells in assay buffer to 3.0 × 10⁶c/ml. inhibitor was diluted to 2 fold final concentration in assay buffer and pre-incubated with HuT78 cells for 30 minutes at room temperature 100. mu.l/well of cells and inhibitor were added to the plate and incubated for 1 hour at room temperature 100. mu.l/well of PBS were added and the plate was sealed, centrifuged at 100g and inverted for 5 minutes, unattached cells were thrown out of the plate and excess PBS was blotted on a paper towel. 60. mu.l/well of p-nitrophenyl n-acetyl- β -D-glucosaminide (0.257g in 100ml citrate buffer) was added to the plate and incubated for 1.5 hours at 37 ℃ the enzyme reaction was stopped with 90. mu.l/well of 50mM glycine/5 mM EDTA and the plate read on a plate reader. the adhesion to cell Ig cell was measured using the method of p-nitrophenyl n-acetyl- β -D-glucosaminide in 37388. U. (1984), J.munol.

T cell proliferation assay:

this assay is an in vitro model of lymphocyte proliferation resulting from activation, induced by engagement of T-cell receptors with LFA-1 upon interaction with antigen presenting cells (Springer, Nature, 346:425 (1990)).

Microtiter plates (Nunc 96 wells, secured by ELISA) were pre-coated overnight at 4 ℃ with 50. mu.l of goat anti-human Fc (Caltag H10700) at 2. mu.g/mL and 50. mu.l of CD3 monoclonal antibody (Immunotech 0178) at 0.07. mu.g/mL in sterile PBS. The next day the coating solution was aspirated. The plate was then washed twice with PBS and 100. mu.l of 17ng/ml 5d-ICAM-1-IgG was added at 37 ℃ for 4 hours. Plates were washed twice with PBS and then CD4+ T cells were added. Lymphocytes from peripheral blood were isolated from heparinized whole blood from healthy donors. Another method is to obtain whole blood from healthy donors by leukapheresis (leukapheresis). Blood was diluted 1:1 with saline, layered and centrifuged at 2500Xg for 30 minutes on LSM (6.2 g Ficoll and 9.4g sodium diztrizoate per 100ml) (Organon Technica, NJ). Monocytes were depleted using the bone marrow cell depletion reagent method (myoclear, Cedarlane Labs, Hornby, Ontario, Canada). PBLs were resuspended in 90% heat-inactivated fetal bovine serum and 10% DMSO, aliquoted and stored in liquid nitrogen. After thawing, cells were resuspended in RPMI 1640 medium (Gibco, Grand Island, NY) supplemented with 10% heat-inactivated fetal bovine serum (Intergen, Purchase, NY), 1mM sodium pyruvate, 3mM L-glutamine, 1mM non-essential amino acids, 500. mu.g/ml penicillin, 50. mu.g/ml streptomycin, 50. mu.g/ml gentamicin (Gibco).

Purification of CD4+ T cells was achieved by negative selection method (human CD4Cell Recovery Column Kit # CL110-5 Accurate). At 37 ℃ in 5% CO₂Next, 100,000 purified CD4+ T cells (90% pure) per microtiter plate well were cultured in 100mL of medium (RPMI 1640(Gibco) supplemented with 10% heat-inactivated FBS (Intergen), 0.1mM non-essential amino acids, 1nM sodium pyruvate, 100 units/mL penicillin, 100. mu.g/mL streptomycin, 50. mu.g/mL gentamicin, 10mM Hepes, and 2mM glutamine) for 72 hours. Inhibitors were added to the plates at the beginning of the culture. Proliferative responses in these cultures were measured by adding 1 μ Ci/well of titrated thymidine during the last 6 hours prior to cell harvest. Radiolabel binding was measured by liquid scintillation counting (Packard 96 well collection device and counter). Results are expressed as counts per minute (cpm).

In vitro mixed lymphocyte culture model:

the mixed lymphocyte culture model is an in vitro model of Transplantation (A.J. Cunningham, "unrestrained Immunology, Transplantation Immunology" pp 157-159 (1978) which examines the proliferation and effector arm effects of different LFA-1 antagonists on human mixed lymphocyte responses.

Isolation of cells: mononuclear Cells (PBMCs) from peripheral blood were isolated from heparinized whole blood from healthy donors. Blood was diluted 1:1 with saline, layered and centrifuged at 2500Xg for 30 minutes on LSM (6.2 g Ficoll and 9.4g sodium diztrizoate per 100ml) (Organon Technica, NJ). Another approach is to obtain whole blood from healthy donors by leukapheresis. PBMCs were isolated as described above, resuspended in 90% heat-inactivated fetal bovine serum and 10% DMSO, aliquoted and stored in liquid nitrogen. After thawing, cells were resuspended in RPMI 1640 medium (Gibco, Grand Island, NY) supplemented with 10% heat-inactivated fetal bovine serum (Intergen, Purchase, NY), 1mM sodium pyruvate, 3mM L-glutamine, 1mM non-essential amino acids, 500. mu.g/ml penicillin, 50. mu.g/ml streptomycin, 50. mu.g/ml gentamicin (Gibco).

Mixed Lymphocyte Response (MLR): one-way human mixed lymphocyte cultures were established in flat-bottomed 96-well microtiter plates. Mix 1.5x10⁵Effector PBMC were co-cultured with an equal amount of stimulator PBMSc irradiated heterologously (3000rads, 3 min 52 sec) in 200. mu.l complete medium. LFA-1 antagonist is added at the beginning of the culture. Cultures were incubated at 37 ℃ in 5% CO₂Medium culture was performed for 6 days, followed by pulse culture with 1. mu. Ci/well of 3H-thymidine (6.7Ci/mmol, NEN, Boston, MA) for 6 hours. Cultures were harvested on a Packard cell harvester (Packard, Canberra, Canada). Measured by liquid scintillation counting³H]TdR binds. Results are expressed as counts per minute (cpm).

Claims (36)

1. An isolated compound represented by structure (I):

and pharmaceutically acceptable salts thereof;

wherein R is¹And R²Each independently is hydrogen; - (CH)₂)_mAn aryl group; in which m is 1- (CH)₂)_mA heteroaryl group; -CH₂NHC (O) heteroaryl; -CH₂NHC(O)NR^1BR^1CWherein R is^1BAnd R^1CIndependently is hydrogen, alkyl, heteroalkyl, cycloalkyl, heterocycle, aryl, heteroaryl, or R^1BAnd R^1CTogether with the nitrogen atom to which they are attached form a heterocyclic or heteroaryl moiety; -CH₂NH heteroaryl; or-CH₂NHC(NR^2ER^2F)＝NR^2CWherein R is^2CIs CN or SO₂R^G2，R^G2Is lower alkyl, and R^2EAnd R^2FEach independently is hydrogen, or an aliphatic, alicyclic, heteroaliphatic, heteroalicyclic, aryl, heteroaryl, alkylaryl, or alkylheteroaryl moiety, or R^2EAnd R^2FTogether form a substituted or unsubstituted heterocyclic or heteroaryl moiety;

R³is-C (═ O) OR^3AWherein R is^3AIs hydrogen;

R^4Aand R^4BIs Cl;

R^B1and R^B2Is hydrogen;

R^Eis hydrogen or C ═ O;

AR¹is that

And

l is C ═ O, -CH₂-, or

2. The compound of claim 1, wherein-C (═ O) NHC (R)¹)(R²)R³Is a moiety having the structure:

wherein Ar is₂Is an aryl or heteroaryl moiety; and R^SIs hydrogen.

3. The compound of claim 1, wherein-C (═ O) NHC (R)¹)(R²)R³Is a moiety having the structure:

wherein R is^1AIs heteroaryl or-NR^1BR^1C(ii) a Wherein R is^1BAnd R^1CIndependently hydrogen, alkyl, heteroalkyl, cycloalkyl, heterocyclic, aryl, heteroaryl, or R^1BAnd R^1CTogether with the nitrogen atom to which they are attached form a heterocyclic or heteroaryl moiety.

4. The compound of claim 1, wherein-C (═ O) NHC (R)¹)(R²)R³Is a moiety having the structure:

wherein Ar is₂Is a heteroaryl moiety; and R^2AIs hydrogen.

5. The compound of claim 1, wherein-C (═ O) NHC (R)¹)(R²)R³Is a moiety having the structure:

wherein R is^2AIs hydrogen;

R^2Cis CN or SO₂R^G2；

R^G2Is a lower alkyl group; and is

R^2EAnd R^2FEach independently hydrogen, or an aliphatic, alicyclic, heteroaliphatic, heteroalicyclic, aryl, heteroaryl, alkylaryl or alkylheteroaryl moietyIs or R^2EAnd R^2FTogether form a substituted or unsubstituted heterocyclic or heteroaryl moiety.

6. The compound of claim 1, wherein-C (═ O) NHC (R)¹)(R²)R^3DIs a moiety having the structure:

wherein R is^2AIs hydrogen;

R^2Eand R^2FEach independently hydrogen, or an aliphatic, alicyclic, heteroaliphatic, heteroalicyclic, aryl, heteroaryl, alkylaryl or alkylheteroaryl moiety, or R^2EAnd R^2FTogether form a substituted or unsubstituted heterocyclic or heteroaryl moiety.

7. The compound of claim 6, wherein R^2EAnd R^2FTogether with the nitrogen atom to which they are attached form a moiety having one of the following structures:

wherein s is an integer from 0 to 6; r^P1Independently at each occurrence is hydrogen, halogen, CN, isocyanate, NO₂、-P(＝O)(YR^P5)₂An alkyl, cycloalkyl, heteroalkyl, heterocyclic, aryl, heteroaryl, alkylaryl or alkylheteroaryl moiety, or is-GR^G1Wherein G is-O-, -S-, -NR^G2-、-CO-、-SO-、-SO₂-、-C(＝O)O-、-C(＝O)NR^G2-、-OC(＝O)-、-NR^G2C (═ O) -or-SO)₂NR^G2-, and R^G1And R^G2Independently is a hydrogen, alkyl, cycloalkyl, heteroalkyl, heterocyclic, aryl, heteroaryl, alkylaryl or alkylheteroaryl moiety; y is independently at each occurrence a bond or O; r^P5Independently at each occurrence is alkyl, heteroalkyl, aryl or heteroaryl, or R when Y is O^P5May also be hydrogen; and R^P2Independently at each occurrence is hydrogen, an aliphatic, alicyclic, heteroaliphatic, heterocyclic, aryl, heteroaryl, alkylaryl, alkylheteroaryl, heteroalkylaryl, or heteroalkylheteroaryl moiety or a nitrogen protecting group; wherein any two adjacent R^P1And R^P2Together may form a cycloalkyl, heterocyclic, aryl or heteroaryl moiety.

8. The compound of claim 6, wherein R^2EAnd R^2FTogether with the nitrogen atom to which they are attached form a moiety having one of the following structures:

wherein R is^P1Independently at each occurrence is hydrogen, halogen, methyl, -OCH₃、-OH、-NH₂、-NHCH₃or-N (CH)₃)₂。

9. The compound of claim 6, wherein R^2EAnd R^2FTogether with the nitrogen atom to which they are attached form a moiety having one of the following structures:

10. the compound of claim 1, wherein-C (═ O) NHC (R)¹)(R²)R³Has the following structure:

wherein R is^P1Independently at each occurrence is hydrogen, halogen, methyl, -OCH₃、-OH、-NH₂、-NHCH₃or-N (CH)₃)₂；R^2AIs hydrogen; and q is 1 or 2.

11. The compound of claim 1, wherein L is-C (═ O).

12. The compound of claim 1, wherein L is-CH₂-。

13. The compound of claim 1, wherein L is a moiety having the structure

14. A pharmaceutical composition comprising a compound according to any one of claims 1 to 13 and a pharmaceutically acceptable carrier or diluent.

15. The pharmaceutical composition of claim 14, wherein the compound is present in an amount effective to modulate adhesion between the intracellular adhesion molecule and the leukocyte integrin family of receptors.

16. The pharmaceutical composition of claim 14, wherein the compound is present in an amount effective to antagonize CD11/CD18 receptors associated with leukocytes.

17. A pharmaceutical composition according to claim 14 wherein the compound is present in an amount effective to antagonize Mac-1 and/or LFA-1.

18. Use of a compound according to any one of claims 1-13 in the manufacture of a medicament for treating an immune or inflammatory response or disorder mediated by the CD11/CD18 family of cell adhesion molecules in a subject.

19. The use of claim 18, wherein the CD11/CD18 family of cell adhesion molecules is LFA-1.

20. Use of a compound according to any one of claims 1-13 in the manufacture of a medicament for treating an immune or inflammatory response or disorder in a subject.

21. The use of claim 20, wherein the disorder is selected from the group consisting of T cell inflammatory responses; responses associated with inflammatory bowel disease; adult respiratory distress syndrome; dermatitis; meningitis; encephalitis; uveitis of the eye; an allergic condition; skin hypersensitivity reactions; atherosclerosis; leukocyte adhesion defects; autoimmune diseases, systemic lupus erythematosus, diabetes, multiple sclerosis, Reynaud's syndrome, autoimmune thyroiditis, experimental autoimmune encephalomyelitis, Sjogren's syndrome, type I diabetes, juvenile diabetes; an immune response associated with delayed hypersensitivity mediated by cytokines and T lymphocytes; pernicious anemia; diseases involving leukocyte extravasation; CNS inflammatory diseases, sepsis or multiple organ damage syndrome secondary to trauma, autoimmune hemolytic anemia, myasthenia gravis, antigen-antibody complex mediated diseases, and transplantation diseases.

22. The use of claim 21, wherein the T cell inflammatory response is an inflammatory skin disease.

23. The use of claim 21, wherein the T cell inflammatory response is psoriasis.

24. The use of claim 21, wherein the inflammatory bowel disease is crohn's disease.

25. The use of claim 21, wherein the inflammatory bowel disease is ulcerative colitis.

26. The use of claim 21, wherein the allergic condition is eczema.

27. The use of claim 21, wherein the allergic condition is asthma.

28. Use according to claim 21, wherein the skin hypersensitivity is poison kudzu skin hypersensitivity.

29. The use of claim 21, wherein the skin hypersensitivity is poison oak skin hypersensitivity.

30. Use according to claim 21, wherein the immune response associated with delayed hypersensitivity mediated by cytokines and T lymphocytes is found in tuberculosis.

31. Use according to claim 21, wherein the immune response associated with delayed hypersensitivity mediated by cytokines and T lymphocytes is found in sarcoidosis.

32. Use according to claim 21, wherein the immune response associated with delayed hypersensitivity mediated by cytokines and T lymphocytes is found in polymyositis.

33. The use of claim 21, wherein the immune response associated with delayed hypersensitivity mediated by cytokines and T lymphocytes is found in granulomatous disease.

34. Use according to claim 21, wherein the immune response associated with delayed hypersensitivity mediated by cytokines and T lymphocytes is found in vasculitis.

35. Use according to claim 21, wherein the transplant disease is graft versus host disease.

36. The use of claim 21, wherein the transplant disease is host anti-transplant disease.