RU2236222C2 - Fc receptor modulators and their application - Google Patents

Abstract

FIELD: medicine, pharmacy.

SUBSTANCE: invention relates to a pharmaceutical composition comprising a compound modulating Fc receptor and pharmaceutically acceptable vehicle. Invention relates also to a method for treatment of different diseases using a compound modulating Fc receptor.

EFFECT: valuable medicinal properties of modulators.

98 cl, 20 dwg, 2 tbl, 26 ex

Description

FIELD OF THE INVENTION

The present invention relates to compounds that modulate the binding of immunoglobulins to Fc receptors and to their use.

BACKGROUND OF THE INVENTION

Fc receptors (FcR) are a family of closely related receptors that are specific for the Fc fragment of an immunoglobulin (Ig). These receptors are essential for normal immunity and resistance to infection and provide the humoral immune system with the capabilities of a cell effector. Receptors are specific for each of the classes of immunoglobulins and are thus characterized by the class of Ig that they bind to (i.e., the Fc gamma receptor (FcgR) binds to gamma immunoglobulin (IgG), the Fc epsilon receptor (FcgR) binds to epsilon immunoglobulin (IgE) , Fc alpha receptor (FcaR) binds to alpha immunoglobulin (IgA)). Among the FcgR receptors, members of three specific subfamilies have been identified; FcgRI, a highly specific receptor for IgG; FcgRII, which are receptors with low affinity for IgG, which vigorously binds to aggregates of immune complexes; and FcgRIII, which are low affinity receptors that bind to immune complexes. These receptors are structurally closely related, but carry out various functions. Of interest is the structure and function of FcgRII due to its interaction with immune complexes and its association with the disease.

FcgRs are expressed by most hematopoietic cells and, by binding to IgG, play a key role in the homeostasis of the immune system and in protecting the host from infection. FcgRII is a receptor with low affinity for IgG and to a large extent binds only to IgG immune complexes and is expressed by cells of various types, including, for example, monocytes, macrophages, neutrophils, eosinophils, platelets and B lymphocytes. FcgRII is involved in various immune and inflammatory reactions, including antibody-dependent cell-mediated cytotoxicity, clearance of immune complexes, isolation of inflammatory mediators, and regulation of antibody formation. The binding of IgG to FcgR can lead to indications in diseases that include the regulation of FcgR. For example, an autoimmune disease, thrombocytopenic purpura, causes tissue (platelet) damage resulting from FcgR-dependent platelet activation of the IgG immune complex or their destruction by FcgR + phagocytes. In addition to this, various inflammatory diseases are known in which IgG immune complexes are involved (for example, rheumatoid arthritis, systemic lupus erythematosis), including allergic reactions of type II and type III. Allergic reactions of type II and type III are carried out with the participation of IgG, which can activate complementary-mediated or phagocytic-effector mechanisms that cause tissue damage.

Due to the fact that FcRs are involved in various biological mechanisms, there is a need for compounds that affect the binding of immunoglobulins to FcRs. There is also a need for the use of such compounds for the treatment of various diseases.

SUMMARY OF THE INVENTION

The present invention provides a pharmaceutical composition comprising:

(a) a compound selected from the group comprising an aromatic compound of the formula:

heteroaromatic compound of the formula:

cyclic compound of the formula:

a bicyclic compound of the formula:

and an amino acid derivative of the formula:

or their salts, where

each of W ¹ and W ² independently represents CO ₂ R ¹⁵ , C (= NH) NH (OH), SO ₃ R ¹⁵ , C (= NH) NH ₂ , OPO (OR ¹⁵ ) ₂ , C (= O) CF ₃ or PO (OR ¹⁵ ) ₂ ;

each of Ar ¹ , Ar ² , Ar ^4, and Ar ⁵ independently is C ₆ -C ₂₀ aryl or C ₁ -C ₂₀ heteroaryl;

Ar ³ is C ₁ -C ₂₀ heteroaryl;

each of X ¹ , X ² , X ³ , X ⁴ , X ⁵ , X ⁶ , X ⁷ and X ⁸ independently represents methylene, O, S or NR ¹⁶ ;

each R ¹ and R ² independently represents a bond, C ₁ -C ₆ alkylene or halogenated C ₁ -C ₆ alkylene;

each R ³ and R ⁴ independently represents halogen, —Z ¹ or C ₁ -C ₆ alkyl;

each X ⁹ , Y ¹ and Z ¹ independently represents OR ¹⁷ , SR ¹⁷ or NR ¹⁷ R ¹⁸ ;

each R ⁵ and R ⁶ independently represents an amino acid residue of a side chain or a particle of the formula —R ¹⁹ —W ³ ;

each R ⁸ , R ⁹ and R ¹¹ independently represents an amino acid residue of the side chain, provided that R ^{11 is} not H or CH ₃ ;

R ⁷ represents OR ²⁰ , NR ²¹ R ^22, or from about 1 to 10 amino acids;

R ¹⁰ represents C ₁ -C ₆ alkylene;

R ¹² represents C ₁ -C ₆ alkyl or C ₆ -C ₂₀ aralkyl;

W ³ represents C (= O) X ¹⁰ ;

X ¹⁰ represents OR ²³ or NR ²⁴ R ²⁵ ;

each R ¹³ , R ¹⁵ , R ¹⁷ . R ¹⁸ , R ²⁰ , R ²¹ , R ²³ and R ²⁴ independently represents hydrogen or C ₁ -C ₆ alkyl;

each R ¹⁶ independently represents H, C ₆ -C ₂₀ aryl or an amide protecting group;

R ¹⁹ represents C ₁ -C ₆ alkylene;

each R ²² and R ²⁵ independently represents H, C ₁ -C ₆ alkyl or an amide protecting group;

R ¹⁴ represents H, C ₁ -C ₆ alkyl or an amine protecting group;

L represents a linking group comprising from 1 to 20 atoms; and

each m and n independently represents an integer from 0 to 2; and

(b) a pharmaceutically acceptable carrier.

The present invention also provides a method of using a compound selected from the group consisting of substituted or unsubstituted benzoic acids; nucleosides and their analogues; folic acid and its derivatives; peptides comprising from about 2 to 10 amino acid residues or their derivatives, preferably tripeptides or hexapeptides; macrocyclic compounds containing a ring that includes from about 8 atoms to 18 atoms, preferably cyclic peptides or derivatives thereof, and compounds with the above formulas for modulating, for example, inhibiting or enhancing the binding of immunoglobulins to patient Fc receptors. In a separate embodiment of the present invention, such modulation of Fc receptors by the aforementioned compounds is used to treat a disease in which antibody aggregates are generated or in which immune complexes are produced by contacting an antibody with an internal or external antigen. Modulation of Fc receptors by the above compounds can also be used to reduce IgG-dependent tissue damage, to reduce the IgE-dependent response and / or to reduce inflammation in a patient.

BRIEF DESCRIPTION OF THE DRAWINGS

In FIG. 1 schematically depicts a binding site at the FcgRIIa receptor.

In FIG. 2 is a side schematic view of a groove; only one surface with protein residues of interest is shown.

In FIG. Figure 3 shows how a single ligand relates to the general structure of the compound of the present invention.

FIG. 4 displays various hydrogen bonds between amino acids at the binding site of an FcgIIa receptor and a specific modulator.

In FIG. 5A and 5B show some compounds modulating Fc receptors, including those corresponding to modulating the Fc receptor shown in FIG. 6-9.

In FIG. 6 shows the modulation activity of some of the compounds shown in FIG. 5A and 5B, binding of FcgRIIa to human IgG1.

In FIG. 7 shows the modulation activity of some of the compounds shown in FIG. 5A and 5B, binding of FcgRIIa to human IgG3.

In FIG. 8 shows the modulation activity of some of the compounds shown in FIG. 5A and 5B, binding of FcgRIIa to human IgG1.

In FIG. 9 shows the modulation activity of some of the compounds shown in FIG. 5A and 5B, binding of FcgRIIa to human IgG3.

In FIG. 10 shows the enhanced binding of sFcgRII to IgG1 and IgG3 in the presence of a hexapeptide.

In FIG. 11 shows the inhibition of the binding of sFcgRII to IgG1 and IgG3 in the presence of a tripeptide.

In FIG. 12 is a graph of transparency versus time versus agonist alone.

In FIG. 13 is a graph of transparency versus time versus presence of agonist and BRI6855 compound.

In FIG. Figure 14 shows a plot of% platelet aggregation at various concentrations of BRI6728.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides various compounds that can modulate the interaction between Fc receptors and immunoglobulins. Out of touch with any of the theories, it is believed that particularly useful compounds interact with region C (see FIG. 1) of an Fc receptor, for example FcgRII. Thus, it is believed that these compounds inhibit the dimerization of two FcgRII proteins, thereby affecting cell signal transmission through one or both of these FcR proteins. In particular, it is believed that peptide residues 117-131 and 150-164 FcgRII form the border region of the FcgIIa dimer, and it is believed that compounds that have an appropriate structure or are able to bind to these regions are good binding modulators. For example, the naturally occurring hexapeptide Phe121 - Ser126 or shorter segments overlap the region of significant interaction due to the presence of hydrogen bonds, and thus are suitable modulators of the dimerization of two FcgRIIa molecules. Such a protein fragment is described as part of SEQ ID No. 3 of US patent application No. 09/245764, “3D Structures and Models of Fc Receptors and Their Use,” filed February 5, 1999.

The compounds of the present invention were obtained by random screening, as well as by targeted drug design, to modulate Fc receptors. FcgRs are expressed by most hematopoietic cells and, by binding to IgG, play a key role in the homeostasis of the immune system and in protecting the host from infection. FcgRII is a low-affinity IgG receptor that binds substantially only to IgG immune complexes and is expressed by cells of various types, including, for example, monocytes, macrophages, neutrophils, eosinophils, platelets and B lymphocytes. FcgRII is involved in various immune and inflammatory responses, including antibody-dependent cell-mediated cytotoxicity, clearance of immune complexes, isolation of inflammatory mediators, and regulation of antibody formation.

The binding of IgG to FcgR can lead to a disease that is associated with regulation by FcgR. For example, an autoimmune disease, thrombocytopenic purpura, causes tissue (platelet) damage resulting from FcgR-dependent platelet activation of the IgG immune complex or their destruction by FcgR + phagocytes. In addition to this, various inflammatory diseases are known in which IgG immune complexes are involved (for example, rheumatoid arthritis, systemic lupus erythematosis), including allergic reactions of type II and type III. Allergic reactions of type II and type III are carried out with the participation of IgG, which can activate complementary-mediated or phagocytic-effector mechanisms that cause tissue damage.

Knowledge of the three-dimensional structure of FcgRIIa or, of course, any FcR can simplify the preparation of therapeutic and diagnostic reagents for the treatment of the disease. For example, knowing the structure of the binding region of FcgRIIa, one can design compounds that can modulate the binding of immunoglobulins to FcgRIIa. The structure of a number of Fc receptors, including FcgRIIa, FceRI and FcgRIIIb, is given in provisional patent application US No. 60/073972, filed February 6, 1998, and in the aforementioned US patent application No. 09/245764, "3D structures and models of Fc receptors and their use ", filed February 5, 1999.

FcgRIIa is a protein dimer and has a C2 axis of symmetry. In FIG. 1 shows a diagram of the structure of the binding region of FcgRIIa according to x-ray diffraction analysis. Out of touch with any of the theories, it is believed that sites A and A ¹ should be areas of interaction with Fc antibodies; thus, a compound that binds or interacts with sites A or A ¹ most likely interferes with the normal binding of this receptor to IgG. In addition, a compound that binds to sites B, C, and / or D can inhibit or facilitate the binding of an antibody if the compound changes the structure of this receptor so that it destabilizes binding to the antibody or promotes dimerization of the receptors, respectively.

In FIG. 2 is a side schematic view of site B, i.e. a groove representing only one surface, with protein residues of interest in the design of the modulator. The edge of the groove contains lysine and histidine residues and is a target for interaction with hydrogen-binding and / or acid groups of a suitable modulator. The surface part of the groove contains a phenylalanine-benzene ring and can be a target during hydrophobic interaction, in particular during pp interactions. The "floor" of the groove contains the residues Phe121, Thr152, Leu159 and Ser161 together with Asn154, Lys117 (frame carbonyl) and Thr119. It is assumed that these proteins are arranged so as to form a pocket that is capable of strong binding to hydrogen and / or Van der Waals interactions with a modulator or ligand.

The features of the grooves described above determine the design and synthesis of the compound, which is generally depicted as follows:

where the "core" is a lipophilic group, such as an aromatic ring, and the "linker" (linking group) is a linked set of from 1 to 20 atoms, preferably from 1 to 10 atoms and more preferably from 2 to 8 atoms. The presence of acid and groups forming a pocket that is directly attached to the linking group is optional. To ensure favorable interaction with major groups, for example Lys117 and His131, at the edge of the groove, acid groups (“acid”) can branch off from the “core” and / or “linking group”. A “pocket” is that part of the molecule that fills the pockets at the bottom of the groove. Alternatively, the modulator may bind or occupy only the receptor pocket. These key points are illustrated by the example in FIG. 3, which shows how a single modulator relates to the overall design presented above, and also are illustrated in FIG. 4, which shows the interaction points between this modulator and the FcgRIIa protein.

An example of a compound containing residues forming a “pocket” is shown in FIG. 3, where a cytosine-like ring is present in the region of the linking group of the compound. Other suitable pocket binder particles include nucleic acids and related structures, such as hydrazides and amidoureas, which are presented below, or their derivatives.

Preferably, these pocket-forming residues are represented by a dimer of the compound, for example, a nucleic acid dimer, hydrazides, amidoureas or their derivatives.

Compounds of the present invention that possess the above basic features include an aromatic compound of the formula:

heteroaromatic compound of the formula:

cyclic compound of the formula:

a bicyclic compound of the formula:

and an amino acid derivative of the formula:

or its salts, where each W ¹ and W ² independently represents CO ₂ R ¹⁵ , C (= NH) NH (OH), SO ₃ R ¹⁵ , C (= NH) NH ₂ , OPO (OR ¹⁵ ) ₂ , C (= O) CF ₃ or PO (OR ^I5 ) ₂ ; each Ar ¹ , Ar ² , Ar ⁴ and Ar ⁵ independently is C ₆ -C ₂₀ aryl or C ₁ -C ₂₀ heteroaryl; Ar ³ is C ₁ -C ₂₀ heteroaryl; each X ¹ , X ² , X ³ , X ⁴ , X ⁵ , X ⁶ , X ⁷ and X ⁸ independently represents methylene, O, S or NR ¹⁶ ; each R ¹ and R ² independently represents a bond, C ₁ -C ₆ alkylene, or halogenated C ₁ -C ₆ alkylene; each R ³ and R ⁴ independently represents halogen, —Z ¹ or C ₁ -C ₆ alkyl; each X ⁹ , Y ¹ and Z ¹ independently represents OR ¹⁷ , SR ¹⁷ or NR ¹⁷ R ¹⁸ ; each R ⁵ and R ⁶ independently represents an amino acid residue of a side chain or a particle of the formula —R ¹⁹ —W ³ ; each R ⁸ , R ⁹ and R ¹¹ independently represents an amino acid residue of the side chain, provided that R ^{11 is} not H or CH ₃ ; R ⁷ represents OR ²⁰ , NR ²¹ R ^22, or from about 1 to 10 amino acid residues; R ¹⁰ represents C ₁ -C ₆ alkylene; R ¹² represents C ₁ -C ₆ alkyl or C ₆ -C ₂₀ aralkyl; W ³ represents C (= O) X ¹⁰ ; X ¹⁰ represents OR ²³ or NR ²⁴ R ²⁵ ; each R ¹³ , R ¹⁵ , R ¹⁷ , R ¹⁸ , R ²⁰ , R ²¹ , R ²³ and R ²⁴ independently represents hydrogen or C ₁ -C ₆ alkyl; each R ¹⁶ independently represents H, C ₆ -C ₂₀ aryl or an amide protecting group; R ¹⁹ represents C ₁ -C ₆ alkylene; each R ²² and R ²⁵ independently represents H, C ₁ -C ₆ alkyl or an amide protecting group; R ¹⁴ represents H, C ₁ -C ₆ alkyl or an amine protecting group; L represents a linking group containing from 1 to 20 atoms; and each m and n independently represents an integer from 0 to 2.

In accordance with the present invention, “alkyl” groups are aliphatic hydrocarbons, which may be straight or branched chain groups. Alkyl groups may optionally be substituted with one or more substituents, such as halogen, alkenyl, alkynyl, aryl, hydroxy, amino, thio, alkoxy, carboxy, oxo or cycloalkyl. Between the alkyl groups one or more oxygen, sulfur, or substituted or unsubstituted nitrogen atoms may be contained. Examples of alkyl groups include methyl, ethyl, isopropyl, n-butyl, tert-butyl, fluoromethyl, difluoromethyl, trifluoromethyl, chloromethyl, trichloromethyl, methoxyethyl, aminomethyl and pentafluoroethyl.

“Aryl” groups are monocyclic or bicyclic carbocyclic or heterocyclic aromatic ring particles. Aryl groups may be substituted with one or more than one substituent, such as halogen, alkenyl, alkyl, alkynyl, hydroxy, amino, thio, alkoxy or cycloalkyl.

“Monoaryl or heteroaryl” refers to a monocyclic carbocyclic or heterocyclic aromatic ring. Examples of monoaryl or heteroaryl rings include pyrrole, thiophene, furan, imidazole, pyrazole, 1,2,4-triazole, pyridine, pyrazine, pyrimidine, pyridazine, thiazole, isothiazole, oxazole, isoxazole, s-triazine and benzene. A preferred group is phenyl.

“Diaryl or heteroaryl” means a bicyclic ring system containing two fused carbocyclic and / or heterocyclic aromatic rings. Examples of diaryl or heteroaryl rings include indene, isoindene, benzofuran, dihydrobenzofuran, benzothiophene, indole, 1H-indazole, indoline, azulene, tetrahydroazulene, benzopyrazole, benzoxazole, benzoimidazole, benzothiazole, 1,3-benzodiazole, 1,3-benzodiazole, 1,3-benzodiazole, 1,3-benzodiazole, 1,3-benzodiazole, 1,3-benzodiazole purine, naphthalene, tetralin, coumarin, chromon, chromene, 1,2-dihydrobenzothiopyran, tetrahydrobenzothiopyran, quinoline, isoquinoline, quinazoline, pyrido [3,4-b] pyridine and 1,4-benisoxazine.

“Aralkyl” refers to an alkyl group substituted with an aryl group. Suitable aralkyl groups include, but are not limited to benzyl, 2-phenylethyl and picolyl. Aryl groups may also be substituted with other suitable functional groups. Aralkyl groups include groups with heterocyclic and carbocyclic aromatic groups.

A “linking group” (L ¹ ) refers to a chain of atoms that links Ar ¹ to Ar ^{2 with a} certain number of atoms. This number, corresponding to the linking group, refers only to the number of atoms that directly bind Ar ¹ and Ar ² . The group L ¹ may contain groups that can participate in the formation of hydrogen bonds and / or in van der Waals interactions with amino acid residues in the groove of the receptor, for example trifluoroacetyl, imide, urea, amidine, amidoxime or their derivatives.

“Amino acid side chain residue” refers to the amino acid side chain that is located at the α-carbon atom of natural and commercially available α-amino acids. Common side chain amino acid residues include hydrogen (glycine), methyl (alanine), -CH ₂ CH ₂ CH ₂ NHC (= NH) NH ₂ (arginine), CH ₂ C (= O) NH ₂ (asparagine), - CH ₂ CO ₂ H (aspartic acid), -CH ₂ CH (cysteine), -CH ₂ CH ₂ C (= O) NH ₂ (glutamine), -CH ₂ CH ₂ CO ₂ N (glutamic acid), -CH ₂ - (4-imidazole) (histidine), -CH (Et) CH ₃ (isoleucine), -CH ₂ CH (CH ₃ ) (leucine), - (CH ₂ ) ₄ NH ₂ (lysine), - (CH ₂ ) ₂ SCH ₃ (methionine), -CH ₂ Ph (phenylalanine), -CH ₂ -CH ₂ -CH ₂ - (proline), -CH ₂ OH (serine), -CH (OH) CH ₃ (threonine), -CH ₂ - (3-indole) (tryptophan), -CH ₂ - (4-hydroxyphenyl) (tyrosine) and -CH (CH ₃ ) ₂ (valine).

The pKa of the corresponding acid groups W ¹ and W ^{2 is} less than 9, more preferably less than 7, and most preferably less than 5. "Corresponding acid groups W ₁ and W ₂ " refer to the parent acid group W ¹ and W ² , for example, in the case of if W ¹ and W ² are esters, the corresponding acid corresponds to carboxylic acid, and if W ¹ and W ² are alkylphosphonates, the corresponding acid corresponds to phosphonic acid. It is important that the pKa of W ¹ and W ² depends not only on the type of W ¹ and W ² , but also on the type of substituents present on the W ¹ and W ² groups and / or in mono- or di-aryl or heteroaryl groups to which W ¹ and W ^{2 are} attached. Thus, for example, the presence of one or more than one electron-delaying group, such as nitro, nitroso, carbonyl, cyano and halo groups, reduces the pKa value of the corresponding W ¹ and W ² acid groups. pKa is defined as -log (Ka), where Ka is the dissociation constant. The value of its (its) dissociation constant indicates the strength of an acid or base in a given medium. For example, strong bases are strong proton acceptors (or electron pair donors) and have high pKa values. The pKa values depend on various factors, such as the nature of the solvent and temperature. For example, water (H ₂ O), but an acid not conjugated to water, which is H ₃ O ⁺ , has a pKa in water of 15.7 at 25 ° C, 16.7 at 0 ° C and 14.7 at 60 ° C In addition to this, in dimethyl sulfoxide (DMSO) at 25 ° C, its pKa is 27.5. As used herein, pKa values refer to pKa values that correspond to a pKa value of water corresponding to about 15.7 unless otherwise specified.

In addition to the formulas given here:

Preferably, W ¹ and W ² independently represent CO ₂ R ⁵ , C (= NH) NH (OH) OPO (OR ⁵ ), C (= O) CF ₃ or PO (OR ⁵ ) ₂ .

Preferably, R ¹ and R ² independently represent a bond, C ₁ -C ₆ alkylene or fluorinated C ₁ -C ₆ alkylene. More preferably, R ¹ and R ² independently are a bond, methylene or difluoromethylene.

Preferably, each Ar ¹ , Ar ² and Ar ⁵ independently is monoaryl or heteroaryl. More preferably, Ar ¹ , Ar ² and Ar ³ would be phenyl.

Preferably, Ar ³ is 2-pyridonyl, and more preferably Ar ³ is 4-Ar ⁴ - (2-pyridonyl), i.e. 2-pyridone at the 4-position attached to the group Ar ⁴ .

Preferably, Ar ⁴ is C ₁ -C ₂₀ heteroaryl. More preferably, Ar ⁴ is pyridyl. Most preferably, Ar ⁴ is 4-pyridyl, i.e. 4-pyridine at the 4-position attached to the group Ar ³ .

Preferably, Y ¹ represents NR ¹⁷ R ¹⁸ . More preferably, Y ¹ represents NH ₂ . Preferably, each R ¹⁵ independently represents hydrogen, methyl or ethyl. Preferably, the group L ¹ is C ₁ -C ₆ alkylene; C ₁ -C ₆ alkenylene, including unsaturated bond groups on the α, β carbon atoms (for example, —CH = CH — C (= O) -); or a group of the formula -R ³³ -X ¹⁴ -, -R ³⁴ -X ¹⁵ -R ³⁵ - or -X ¹⁶ -R ³⁶ -Ar ⁷ -R ³⁷ -X ¹⁷ -. Each of R ³³ , R ³⁴ , R ³⁵ , R ³⁶ and R ³⁷ independently represents C ₁ -C ₆ alkylene (including substituted alkylene), preferably methylene. Each X ¹⁴ , X ¹⁵ , X ¹⁶ and X ¹⁷ independently represents O, S or NR ³⁸ , preferably O or NR ³⁸ . Each Ar ⁶ and Ar ⁷ independently represents C ₆ -C ₂₀ aryl or C ₁ -C ₂₀ heteroaryl, preferably 2-pyridone. And R ³⁸ is H, C ₁ -C ₆ alkyl or an amine protecting group, preferably —CH ₂ CO ₂ N. More preferably, L ¹ is sulfonamide (—SO ₂ NH—), ethylene (—CH ₂ CH ₂ -), -CH ₂ O-, -CH = CHS (= O) -, -CH ₂ CH ₂ CH (OH) -, -CH = CH-, -CH (OH) CH (OH) -, -CH ₂ N (R ³⁸ ) CH ₂ -, a group of the formula:

or a group of the formula:

where each R ²⁷ and R ²⁸ independently represents H, C ₁ -C ₆ alkyl, C ₆ -C ₁₀ aralkyl or a protective group. Preferred R ²⁷ and R ²⁸ independently represent H or a protective group. More preferably, R ²⁷ and R ^{28 are} independently H or 4-methoxybenzyl.

Preferably m and n are 0.

Alternatively, R ¹ and W ¹ and / or R ² and W ² together form - (CH ₂ ) _a CH (NHR ²⁹ ) CO ₂ R ³⁹ and - (CH ₂ ) _b CH (NHR ³⁰ ) CO ₂ R ^40, respectively where a and b independently represent an integer from 0 to 2, R ²⁹ and R ³⁰ independently represent H or an amine protecting group, and R ³⁹ and R ⁴⁰ independently represent H or C ₁ -C ₆ alkyl. Preferably, a and b would be 1. Preferably, R ²⁹ and R ³⁰ independently represent H, C ₁ -C ₆ alkyl or an amine protecting group.

Preferably, R ⁵ is an asparagine side chain residue.

Preferably, R ⁶ is a glutamine side chain residue.

Preferably, R ⁷ represents from about 1 to 10 amino acid residues or their derivatives, more preferably from 1 to 6 amino acid residues or their derivatives, even more preferably at least 2 amino acid residues or their derivatives, and most preferably the particle -lys- ser-CONHCH ₃ , i.e. a particle of the formula NHCH [(CH ₂ ) ₄ NH ₂ ] CONHCH (CH ₂ OH) CONHCH ₃ .

Preferably, X ¹ , X ² , X ³ , X ⁴ , X ⁵ , X ⁶ , X ⁷ and X ⁸ independently represent O or NR ¹⁶ . More preferably, X ¹ , X ² , X ³ , X ⁴ , X ⁵ , X ⁶ , X ^7, and X ⁸ are NR ¹⁶ .

Preferably, X is OR ¹⁷ or NR ¹⁷ R ¹⁸ , more preferably NR ¹⁷ R ^18, and most preferably NH ₂ .

Preferably, R ⁸ is a glycine side chain residue (i.e., H).

Preferably, R ⁹ is a tyrosine side chain residue (i.e. 4-hydroxybenzyl).

Preferably, R ¹⁰ is propylene.

Preferably, R ¹¹ is a lysine side chain residue, i.e. a particle of the formula - (CH ₂ ) ₄ NH ₂ .

Preferably, R ¹² is C ₆ -C ₂₀ aralkyl, and more preferably 2-phenylethyl.

Preferably, R ¹³ represents N.

Preferably, R ¹⁴ is H or an amine protecting group, more preferably an amine protecting group, and most preferably an acetyl group, i.e. a group of the formula —C (═O) CH ₃ .

Preferably, each R ¹⁶ independently represents H or C ₆ -C ₂₀ aryl. More preferably, each R ¹⁶ independently represents H or phenyl.

In one particular embodiment of the present invention, the above aromatic compound is a compound of the formula:

More preferably, this aromatic compound would be a compound of the formula:

In another particular embodiment of the present invention, the aromatic compound described above is a compound of the formula:

In one particular embodiment of the present invention, the heteroaromatic compound described above is a compound of the formula:

More preferably, this heteroaromatic compound described above is a compound of the formula:

In another particular embodiment of the present invention, the cyclic compound described above is a compound of the formula:

In another particular embodiment of the present invention, the bicyclic compound described above is a compound of the formula:

In another particular embodiment of the present invention, the amino acid derivative described above is a derivative of the formula:

or its salts. Preferably, the amino acid derivative described above is a derivative of the formula:

or was its salt.

The Fc receptor modulating compounds of the present invention may also include nucleosides or their derivatives. Preferably, the nucleosides of the present invention would have the formula:

where Q represents O or methylene. More preferably Q is O. X ¹¹ is OR ³¹ or OPO (OR ³¹ ). More preferably X ¹¹ is OH or OPO ₃ H ₂ . Each X ¹² and X ¹³ independently represents H or OR ¹⁵ . Preferably, each X ¹² and X ¹³ independently represents H or OH. Each R ³¹ and R ³² independently represents H or C ₁ -C ₆ alkyl.

The Fc receptor modulating compounds of the present invention may also include folic acid or its derivatives.

The Fc receptor modulating compounds of the present invention may also include peptides that can modulate the interaction between Fc receptors and immunoglobulins. Out of touch with any of the theories, it is believed that particularly useful peptides interact with region C (see FIG. 1) of Fc receptors, for example FcgRII. Thus, it is believed that these peptides inhibit dimerization between two FcgRII proteins, thereby affecting signal transmission from cells through one or both FcR proteins. In this case, residues 117-131 and residues 150-164 form the boundary region of the FcgIIa dimer, and peptides with such sequences or with such properties are binding inhibitors. For example, the naturally occurring hexapeptide Phe121 - Ser126 or shorter segments overlap a region with a large number of hydrogen bonds causing a significant interaction, and thus are suitable modulators of the dimerization of two FcgRIIa molecules. Such a protein fragment is described as part of SEQ ID NO: 3 in the aforementioned US patent application No. 09/245764, “3D Structures and Models of Fc Receptors and Their Use,” filed February 5, 1999. Thus, the present researchers found that tripeptide c the sequence of GKS (gly-lys-ser) or its derivatives and hexapeptides with the sequence FQNGKS (phe-gln-asn-gly-lys-ser) or their derivatives modulate the binding of FcgRII to IgG. See experiment 24 and FIG. 10 and 11.

The present researchers also found that macrocyclic compounds with a specific conformation modulate the behavior of the FcR protein. As used herein, the term “macrocyclic compound” refers to a compound containing a ring that includes from 8 atoms to 18 atoms. Preferably, the ring of the macrocyclic compound of the present invention comprises from about 10 to 16 atoms, more preferably from 12 to 14 atoms, and most preferably from 13 to 14 atoms. A particularly useful macrocyclic compound of the present invention is a cyclic peptide or its derivatives. Such a cyclic peptide having the formula:

was described above. The top of the FG cycle of the FcR was determined in mutagenesis studies that are relevant in the study of Ig binding. The FG peptide region contains an expanded b-fold in which the amino acid side residues of the FG loop are oriented at a specific position. This orientation of the Fc protein is described in the aforementioned US patent application No. 09/245764, "3D Structures and Models of Fc Receptors and Their Use", filed February 5, 1999. It has also been found that molecules that can reproduce turns of the b-helix so that its lateral residues occupy a position in the upper part of the FG loop, i.e. just as is the case for the lateral residues of the receptor, they effectively modulate the behavior of the FcR receptor. Thus, in yet another embodiment of the present invention, the Fc receptor modulating compound is also a compound of the formula:

where the macrocyclic fragment contains the same number of atoms as described above. One particular embodiment of such a b-helix loop reproduction is a compound described above, which has the formula:

The compounds of the present invention can be synthesized from readily available starting materials. Various substituents in the compounds of the present invention may be present in the starting compounds, introduced into any of the intermediates, or introduced after the formation of the final products by known substitution or conversion methods. If the substituents themselves are reactive, then these substituents can be protected in accordance with known methods. Various protecting groups are known that can be used. Examples of many possible groups can be found in the publication "Protective Groups in Organic Synthesis" by T.W. Green, John Wiley and Sons, 1981. For example, nitro groups can be introduced by nitration, and the nitro group can be converted to other groups, such as the amino group by reduction and the halogen group by diazotization of the amino group and the replacement of this diazo group with halogen. Acyl groups can be introduced by Friedel-Crafts acylation. These acyl groups can then be converted to the corresponding alkyl groups in various ways, including Wolf-Kizhner reduction and Clemenson reduction. Amino groups can be alkylated to form mono- and di-alkylamino groups; and mercapto and hydroxy groups can be alkylated to form the corresponding esters. Primary alcohols can be oxidized by known oxidizing agents to form carboxylic acids or aldehydes, and secondary alcohols can be oxidized to form ketones. Thus, substitution or change reactions can be used to introduce various substituents into the molecule of the starting material, into the intermediates, or into the final product, including the isolated products. Since the compounds of the present invention may have some substituents that must be present, the introduction of each substituent, of course, depends on the nature of the substituents involved and the chemistry of the reactions necessary for their formation. Thus, the solution of the question of how the chemical reaction will influence a separate substituent during the formation of the second substituent will be associated with methods that are familiar to the average person. It will also depend on the nature of the ring.

It should be borne in mind that the present invention covers not only various isomers that may exist, but also various mixtures of isomers that may be formed.

If the compound of the present invention contains one or more chiral centers, it can be synthesized enantioselectively or as a mixture of enantiomers, and / or diastereomers can be obtained, followed by resolution. The separation of the compounds of the present invention, the starting materials for their preparation and / or intermediates can be obtained by known methods, for example, as described in the four-volume Optical Resolution Procedures for Chemical Compounds: Optical Resolution Information Center, Manhattan College, Riverdale, NY, and Enantiomers, Racemates and Resolution, Jean Jacques, Andre Collet and Samuel H. Wilen; John Wiley & Sons, Inc., New York, 1981. In general, the separation of compounds is based on differences in the physical properties of the diastereomers upon chemical or enzymatic addition of an enantiomerically pure group to form forms that are separated by fractional crystallization, distillation, or chromatography.

If the compound of the present invention contains an olefin group and this group can have either a cis or trans configuration, then the compound can be synthesized to form either a cis or trans olefin as the predominant product. Alternatively, a compound containing an olefin group can be obtained as a mixture of cis and trans olefins and separated by known methods, for example, chromatography as described in W.K. Chan, et al., J. Am. Chem. Soc., 1974, 96, 3642.

The compounds of the present invention form salts formed by addition of acids in case a basic amino function is present, and salts formed by addition of bases if an acid function is present, for example carboxylic acid or phosphonic acid. All such salts are useful in isolating and / or purifying new products. Of particular importance are pharmaceutically acceptable salts with both acids and bases. Suitable acids include, for example, hydrochloric, oxalic, sulfuric, nitric, benzenesulfonic, toluenesulfonic, acetic, maleic, tartaric and the like acids, which are pharmaceutically acceptable. Basic salts for pharmaceutical use include salts of Na, K, Ca and Mg.

In addition to and / or instead of targeted drug development, other Fc receptor modulators can be identified by a screening method in which various compounds are tested to determine their activity in modulating the Fc receptor. In this way, various Fc receptor modulators have been identified. Thus, the compounds of the present invention include substituted and unsubstituted benzoic acids, in particular 4-methylbenzoic acid and 3-methylbenzoic acid; nucleosides and their analogues; as well as folic acid and its derivatives.

The compounds of the present invention are modulators of the Fc receptor, for example, they modulate the binding of the Fc receptor to immunoglobulins. Preferably, the compounds of the present invention would modulate Fc receptors selected from the group consisting of FcaR, FcgR, FcgR and mixtures thereof, more preferably from the group comprising FcgRI, FcgRII, FcgRIII and mixtures thereof, even more preferably from the group including FcgRIIa, FcgRIIb, FcgRIIc, and mixtures thereof, and most preferably the FcgRIIa receptor. The compounds of the present invention can be used for a variety of purposes, including the treatment or diagnosis of any disease in which aggregates with antibodies are formed and in which immune complexes are formed when the antibody contacts an internal or external antigen. Examples of the use of the compounds of the present invention for treatment and diagnosis include immune complex diseases; autoimmune diseases, including but not limited to rheumatoid arthritis, systemic lupus erythematosis, immune thrombocytopenia, neutropenia, hemolytic anemia; vasculitis, including but not limited to polyarthritis thickening, systemic vasculitis; xenograft rejection; and infectious diseases in which FcR absorbs a viral infection, including but not limited to flavivirus infections, such as dengue virus hemorrhagic fever, and measles viral infection. The compounds of the present invention can also be used to reduce IgG-mediated tissue damage and to reduce inflammation.

The compounds of the present invention can also improve leukocyte function as a result of improved FcR function. These functions include antibody-dependent cell-mediated cytotoxicity, phagocytosis, and the release of inflammatory cytokines. Examples of treatment and diagnosis for improving FcR function include any infection in which normal antibodies are produced to eliminate the pathogenic factor; and any disease requiring an FcR function in which natural or recombinant antibodies can be used to treat diseases such as cancer and infections, for example, antibodies can be administered together with the compound of the present invention to improve the effect of antibody treatment.

The compounds of the present invention can be administered to a patient to achieve the desired physiological effect. Preferably, the patient is an animal, more preferably a mammal, and most preferably a human. The compound may be administered in various forms intended for the chosen route of administration, i.e. orally or parenterally. In this regard, parenteral administration includes administration in the following ways: intravenously; intramuscularly; subcutaneously; intraocular, intrasinovial; transepithelial, including transdermal, ophthalmic, sublingual and buccal; local administration, including ophthalmic, dermal, ocular, rectal and nasal inhalations using insufflation and aerosol; intraperitoneally; and rectally systematically.

The active compound may be administered orally, for example, with an inert diluent or with an assimilable edible carrier, or it may be enclosed in hard or soft gelatin capsules, or it may be compressed into tablets, or it may be incorporated directly into food or diet. For oral therapeutic administration, the active compound may be incorporated into an excipient and used in the form of swallowable tablets, buccal tablets, lozenges, capsules, elixirs, suspensions, syrups, wafers and the like. Such compositions and pharmaceutical preparations may contain at least 0.1% of the active compound. The percentage of compositions and pharmaceutical preparations may, of course, vary and may comprise from about 1 to 10% of the weight of the form. The amount of active compound in such therapeutically useful compositions is such that a suitable dose is achieved. Preferred compositions or pharmaceutical preparations in accordance with the present invention are prepared so that the oral dosage form contains from 1 to 1000 mg of the active compound.

Tablets, lozenges, pills, capsules and the like may also contain the following: a binder, such as tragacanth gum, gum arabic, corn starch or gelatin; excipients such as dicalcium phosphate; baking powder, such as corn starch, potato starch, alginic acid and the like; a lubricant such as magnesium stearate; and a sweetener, such as sucrose, lactose or saccharin, or a flavoring agent, such as peppermint oil, gaulteria oil, or cherry flavoring, may be added. If the dosage form is a capsule, then it may contain, in addition to the materials of the above types, a liquid carrier. Various other materials can be used as coatings or to modify the physical form of the drug. For example, tablets, pills or capsules may be coated with shellac, sugar, or both. A syrup or elixir may contain an active compound, sucrose as a sweetener, methyl and propyl parabens as preservatives, a coloring agent and a flavoring agent such as cherry or orange flavoring. Of course, any materials used in the manufacture of any dosage form must be pharmaceutically pure and non-toxic in the quantities used. In addition to this, the active compound may be included in pharmaceutical preparations and long-acting formulations.

The active compound may also be administered parenterally. Solutions of the active compound in the form of a free base or a pharmacologically acceptable salt may be prepared in water suitably mixed with a surfactant such as hydroxypropyl cellulose. Dispersions can also be prepared on glycerol, liquid polyethylene glycols and their mixtures and on oils. Under ordinary conditions of storage and use, these pharmaceutical preparations contain a preservative to prevent the growth of microorganisms.

Pharmaceutical formulations suitable for use as an injection include sterile aqueous solutions or dispersions and sterile powders for the preparation of sterile injectable solutions or dispersions for immediate administration. In all cases of administration using a conventional syringe, this form must be sterile and liquid. It must be stable under the conditions of receipt and storage and must be protected from the contaminating effects of microorganisms such as bacteria and fungi. The carrier may be a solvent in the form of a dispersion medium containing, for example, water, ethanol, a polyol (e.g. glycerin, propylene glycol, as well as liquid polyethylene glycol and the like), suitable mixtures thereof and vegetable oils. Proper fluidity can be achieved, for example, by the use of a coating such as lecithin, the establishment of the required particle size in case of dispersion, and the use of surfactants. Prevention of exposure to microorganisms can be carried out using various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it is preferable to include isotonic agents, for example, sugars or sodium chloride. The prolongation of the absorption of the injectable compositions is carried out using agents that prolong the absorption time, for example, aluminum monostearate and gelatin.

Sterile injectable solutions are prepared by introducing the active compound into the appropriate solvent in the required amount, which contains various other ingredients listed above, as appropriate, followed by sterilizing filtration. In general, dispersions are prepared by incorporating various sterile active ingredients into a sterile carrier that contains a basic dispersing medium and other necessary ingredients from the above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and freeze drying methods, which make it possible to obtain an active ingredient powder containing any additionally required ingredient from their solution that has previously been sterilized by filtration.

The therapeutic compounds of the present invention can be administered to mammals alone or in conjunction with the pharmaceutically acceptable carriers mentioned above, the ratios of which are determined by the solubility and chemical properties of the compound selected by the administration and standard pharmaceutical practice.

The attending physician determines the dosage of the present therapeutic agents, which will be most suitable for prevention or treatment and will vary depending on the form of administration and the particular compound selected, and will also vary during treatment depending on the characteristics of a particular patient. Typically, the attending physician wishes to start treatment with small doses, gradually increasing them until, under the circumstances, the optimal effect is achieved. The therapeutic dose can usually be from 0.1 to 1000 mg / day, and preferably from 10 to 100 mg / day, or from 0.1 to 50 mg / kg of body weight per day, and preferably from 0.1 to 20 mg / kg of body weight per day and can be administered in the form of several different dosage units. When administered orally, 2 to 4 times larger dosages may be required.

Additional objectives, advantages and new features of the present invention will be apparent to those skilled in the art when studying the examples below, which do not limit it.

EXPERIMENTAL DATA

The following abbreviations are used in this document:

c.t. - room temperature,

Et ₂ O is diethyl ether (i.e. ether or ethyl ether),

MS (HIAD) - chemical ionization at atmospheric pressure,

THF - tetrahydrofuran,

EtOAc - ethyl acetate,

TMCCl - trimethylsilyl chloride,

CH ₃ CN - acetonitrile,

DMF - dimethylformamide.

Experience 1

In this experiment, a synthesis of 1,2-bis (m-carboxyphenyl) ethane is presented:

Stage 1: 1,2-bis (m-bromophenyl) ethane obtained by the method of Lindsay et al (JACS, 1961, 83, 943) as follows. To a solution of 3-bromobenzyl bromide (1.0 g, 4.0 mmol) in Et ₂ O (10 ml) at rt magnesium is added (0.05 g, 2.0 mmol). After 20 minutes at room temperature, all of the magnesium dissolves, and then anhydrous ferric chloride (5 mg) is added. The reaction mixture is heated to reflux for 1 hour, cooled, acidified to about pH 1 1 M aqueous H ₂ SO ₄ and extracted with Et ₂ O (3 × 50 ml). The combined organic extracts were washed with water (50 ml), dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo to give a yellow solid. Recrystallization from petroleum ether gave 1,2-bis (m-bromophenyl) ethane as a colorless solid. MS (HIAD) m / z 338 (50%), 340 (100), 342 (50). ¹ H NMR (200 MHz, CDCl ₃ ): d 2.85, s, 2H; 7.02-7.25, m, 2H; 7.30-7.39, m, 2H.

Stage 2: tert-butyl lithium (2.1 ml of a 1.7 M solution in pentane, 3.60 mmol) is added dropwise to a solution of 1,2-bis (m-bromophenyl) ethane (305 mg, 0.90 mmol) in THF (10 ml) at -78 ° C. After 20 minutes at this temperature, a mixture of CO _{2 is} bubbled through the reaction mixture until the temperature of the reaction mixture in the absence of a cooling bath reaches rt. This reaction mixture was partitioned between water (50 ml) and Et ₂ O (50 ml), the aqueous phase was separated and acidified to approximately pH 1 with concentrated aqueous HCl, keeping the internal temperature below 25 ° C. The aqueous phase was extracted with EtOAc (3 × 50 ml) and the combined organic extracts were dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo to give 1,2-bis (m-carboxyphenyl) ethane as a white solid. MS (CHIAD) m / z 269 (M + 1, 100%). ¹³ C NMR (50 MHz, d ₆ -DMSO): d 38.4, 128.8, 130.3, 131.1, 132.5, 134.8, 143.5, 169.2. The melting point is consistent with the value given by Lindsay et al. (JACS, 1961, 83, 943).

Experience 2

In this experiment, the synthesis of 3 - [(m-carboxyphenyl) methoxy] benzoic acid is presented:

Stage 1: a mixture of 3-bromophenol (13.8 g, 80 mmol), 3-bromobenzyl bromide (10 g, 40 mmol), K ₂ CO ₃ (16.6 g, 120 mmol) and NaI (300 mg, 2 mmol) in acetone (100 ml) is heated to reflux for 12 hours. The reaction mixture was cooled to rt, concentrated in vacuo, and partitioned between Et ₂ O (300 ml) and water (300 ml). The organic phase is washed with an aqueous NaOH solution (1 M, 300 ml), dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo to give 3 - [(m-bromophenyl) methoxy] bromobenzene as a clear oil. MS (HIAD) m / z 339 (M ⁺ -3.50%), 341 (M ⁺ -1, 100%), 343 (M ⁺ + 3.50%). ¹³ C NMR (50 MHz, CDCl ₃ ); d 68.9, 113.4, 117.9, 122.5, 122.6, 124.1, 125.6, 129.9, 130.0, 130.4, 130.9, 138.4, 158 ,9.

Step 2: Using 3 - [(m-bromophenyl) methoxy] bromobenzene and using the method described in Example 1, step 2, 3 - [(m-carboxyphenyl) methoxy] benzoic acid is obtained as a white solid. MS (HIAD) m / z 271 (M ⁺ -1, 100%). ¹³ C NMR (50 MHz, d ₆ -DMSO): d 68.3, 114.5, 119.3, 121.5, 127.8, 128.3, 129.3, 130.5, 131.5, 131.8, 137.0, 157.7, 166.6, 166.7.

Experience 3

In this experiment, the synthesis of 1,2-bis (3-phosphonophenyl) ethane is presented:

Stage 1: 1,2-bis (3-bromophenyl) ethane (obtained using the method of Example 1, stage 1) (440 mg, 1.29 mmol), diethyl phosphite (0.46 ml, 3.59 ml) and triethylamine ( 0.5 ml, 3.59 mmol) is dissolved in toluene and degassed. Then, Pd (PPh ₃ ) ₄ (185 mg, 0.16 mmol) was added in one portion, and the reaction mixture was heated to 90 ° C. for 16 hours. The reaction mixture was cooled to room temperature and purified by column chromatography (SiO ₂ , 50% EtOAc in petroleum ether ® 100% EtOAc ® 100% EtOH) to give 1,2-bis [3- (diethoxyphosphono) phenyl] ethane as a white solid substances. MS (HIAD) m / z 455 (M ⁺ +1, 100%). ³¹ P NMR (81 MHz, proton decoupling, CDCl ₃ ): d +19.5.

Step 2: Trimethylsilyl bromide (1.03 ml, 7.8 mmol) is added dropwise to a solution of the above ether (586 mg, 1.30 mmol) in CH ₂ Cl ₂ (10 ml) at rt. The reaction mixture was stirred for 16 hours at room temperature and concentrated in vacuo. MeOH (5 ml) was then added, and this solution was concentrated in vacuo. This operation is repeated two more times to give 1,2-bis (3-phosphonophenyl) ethane as a white solid. MS (HIAD) m / z 341 (M ⁺ -1, 100%). ³¹ P NMR (81 MHz, proton decoupling, CDCl ₃ ): d +14.6.

Experience 4

In this experiment, a synthesis of 3,3’-dicarboxy-chalcone is presented:

Stage 1: 3-Cyanobenzaldehyde (3.0 g, 23.0 mmol), 3-cyanoacetophenone (3.34 g, 23.0 mmol) in glacial acetic acid (5 ml) and concentrated H ₂ SO ₄ (3.66 ml, 69 mmol) was stirred at room temperature for 72 hours. Water (200 ml) was then added and the reaction mixture was filtered. The precipitate was washed with water (2 × 200 ml) and dried in vacuo to give 3.3'-dicyanochalcone as an off-white solid. MS (HIAD) m / z 258 (M ⁺ -1, 100%). ¹³ C NMR (50 MHz, d ₆ -DMCO): d 111.7, 117.8, 118.0, 123.0, 129.7, 131.6, 132.1, 132.4, 133.3, 133.5, 135.3, 136.1, 137.4, 142.1, 187.3.

Stage 2: The solution of 3,3'-dicyanohalcon obtained in stage 1 (2.0 g, 7.75 mmol) in glacial acetic acid (30 ml) is treated with a mixture of concentrated H ₂ SO ₄ (10 ml) and water (10 ml). This reaction mixture was heated to 130 ° C for 12 hours, cooled to room temperature and filtered. The precipitate was washed with water (3 × 100 ml) and dried in vacuo to give 3.3'-dicarboxychalcone as a yellow solid. MS (HIAD) m / z 295 (M ⁺ -1, 100%). ¹³ C NMR (50 MHz, d ₆ -DMSO); d 122.5, 128.6, 128.7, 129.2, 130.8, 131.0, 131.2, 132.4, 132.5, 133.1, 134.5, 137.2, 143 , 1, 166.3, 166.5, 188.2.

Experience 5

In this experiment, a synthesis of 1,3-bis (m-carboxyphenyl) -1-propanol is presented:

3.3'-dicarboxychalcon (Example 4, step 2) (430 mg, 1.45 mmol) in ethanol (10 ml) containing an aqueous solution of NaOH (1 M, 2.90 mmol) was hydrogenated at a pressure of 310 kPa for 48 hours in the presence of a Wilkinson catalyst (67 mg, 0.07 mmol). The reaction mixture was filtered and concentrated in vacuo. The residue was dissolved in methanol (10 ml) and treated with NaBH ₄ (220 mg, 5.8 mmol) at rt. The reaction mixture was stirred for 16 hours at rt, quenched by careful addition of a saturated aqueous solution of NH ₄ Cl and partitioned between EtOAc (50 ml) and aqueous HCl (1 M, 50 ml). The organic extract was dried (Na ₂ SO ₄ , filtered and concentrated in vacuo to give 1,3-bis (m-carboxyphenyl) -1-propanol as a viscous oil. MS (HIAD) m / z 299 (M ⁺ -1, 100 %). ¹ H NMR (200 MHz, CDCl ₃ ); d 1.95-2.10, m, 2H; 2.68-2.83, m, 2H; 4.62-4.78, m, 1H ; 7.03-7.60, m, 4H; 7.75-8.03, m, 4H.

Experience 6

In this experiment, a synthesis of trans-3,3’-bis-carboxystilbene is presented:

Stage 1: Methyl 3-bromobenzoate (21.5 g, 100 mmol), Pd (OAc) ₂ (224 mg, 1 mmol), tri-o-tolylphosphine (608 mg, 2 mmol) and tributylamine (26.2 ml, 110 mmol) in DMF (100 ml) is degassed with argon, heated to 130 ° C for 6 hours, and ethylene is bubbled through this solution. The reaction mixture was cooled to room temperature and filtered. The precipitate was washed with cold Et ₂ O (2 × 50 ml) and dried in vacuo to give trans-3,3'-bis-carboxystilbene dimethyl ether as an off-white solid. ¹³ C NMR (50 MHz, CDCl ₃ ): d 52.2, 127.5, 128.8, 130.6, 130.9, 137.2, 166.9.

Stage 2: The above diester (500 mg, 1.7 mmol) in THF (10 ml) is treated at room temperature with an aqueous solution of LiOH (1 M, 10 ml). After stirring for 16 hours at rt the reaction mixture was partitioned between Et ₂ O (50 ml) and water (50 ml). The aqueous phase was separated and the organic phase was extracted with water (25 ml). The combined aqueous extracts are acidified with concentrated aqueous HCl (while keeping the internal temperature below 10 ° C). The aqueous phase was extracted with CH ₂ Cl ₂ (3 × 50 ml) and the combined organic extracts were dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo to give trans-3,3'-biscarboxystilbene as a white solid. MS (HIAD) m / z 267 (M ⁺ -1, 100%). ¹ H NMR (200 MHz, d ₆ -DMSO): d 7.28-7.56, m, 2H; 7.78-7.90, m, 2H; 8.20, s, 1H.

Experience 7

In this experiment, the synthesis of (S, S) -1,2-bis- (3-carboxyphenyl) ethane-1,2-diol is presented:

Stage 1: Trans-3,3'-biscarboxystilbene dimethyl ester (Example 6, stage 1) (5.0 g, 16.9 mmol) and N-methylmorpholine-N-oxide (2.2 g, 18.6 mmol) in acetone (50 ml) and water (20 ml) is treated at room temperature with an aqueous solution of OsO ₄ (4.3 ml, 39.4 mmol, 0.17 mmol). The reaction mixture was stirred for 16 hours at rt, quenched by the addition of sodium metabisulfite (3.0 g) and adjusted to about pH 7 with a 2 M aqueous sulfuric acid solution. The acetone was removed in vacuo, and the remaining solution was acidified to about pH 2, saturated with NaCl and extracted with EtOAc (3 × 100 ml). The combined organic extracts were dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo to give (R, R) -1,2-bis- [3- (carbomethoxy) phenyl] ethane-1,2-diol as a white solid substances. ¹ H NMR (200 MHz, CDCl ₃ ): d 3.2, bs, 1H; 3.82, s, 3H; 4.77, s 1H; 7.20-7.31, m, 2H; 7.80-7.89, m, 2H.

Stage 2: The above diester (500 mg, 1.5 mmol) is hydrolyzed using the procedure described in Example 6, stage 2, to obtain (S, S) -1,2-bis- (3-carboxyphenyl) ethane-1, 2-diol as a white solid. MS (HIAD) m / z 301 (M ⁺ -1, 100%). ¹ H NMR (200 MHz, d ₆ -DMSO): d 3.40, bs, 1H; 4.76, s, 1H; 5.56, bs, 1H; 7.20-7.29, m, 2H; 7.80-7.91, m, 2H.

Experience 8

In this experiment, a synthesis of 3,3'-bis- (carboxymethyl) stilbene is presented:

Step 1: Methyl 3-bromophenyl acetate (8.0 g, 34.9 mmol) was reacted with ethylene using the procedure described in Example 6, step 1. The crude reaction product was purified by column chromatography (SiO ₂ , 5% EtOAc in petroleum ether ) to give 3,3'-bis - [(carbo-methoxy) methyl] stilbene and methyl 3- (ethenyl) phenyl acetate as a white solid.

3,3'-bis - [(carbo-methoxy) methyl] stilbene: ¹ H NMR (200 MHz, CDCl ₃ ): d 3.65, s, 2H; 3.70, s, 3H; 7.1, s, 1H, 7.15-7.50, m, 4H. ¹³ C NMR (50 MHz, CDCl ₃ ): d 41.2, 52.1, 125.3, 127.4, 128.6. 128.7, 128.9, 134.4, 137.6, 171.9.

Methyl 3- (ethenyl) phenylacetate: ¹ H NMR (200 MHz, CDCl ₃ ): d 3.63, s, 2H; 3.68, s, 3H; 5.28, d, J = 10.9 Hz, 1H; 5.78, d, J = 18.8 Hz, 1H; 6.72, d, J = 10.9, 18.8 Hz, 1H; 7.18-7.41, m, 4H.

Stage 2: 3,3'-bis - [(carbomethoxy) methyl] stilbene is hydrolyzed using the procedure described in Example 6, stage 2, to obtain 3,3'-bis - [(carboxy) methyl] stilbene as a white solid substances. MS (HIAD) m / z 295 (M ⁺ -1, 100%). ¹ H NMR (200 MHz, d ₆ -DMCO): d 3.60, s, 2H; 7.00-7.62, m, 5H.

Experience 9

In this experiment, the synthesis of 1,2-bis- [m- (carboxymethyl) phenyl] ethane is presented:

Stage 1: 3,3'-bis - [(carbomethoxy) methyl] stilbene (Example 8, stage 1) (500 mg, 1.5 mmol) and palladium on carbon (10%, 200 mg) in methanol (20 ml) hydrogenated in a hydrogen atmosphere for 16 hours at rt The reaction mixture was filtered and concentrated in vacuo to give 1,2-bis- [m- (carbomethoxymethyl) phenyl] ethane as a colorless oil. ¹ H NMR (200 MHz, CDCl ₃ ): d 2.91, s, 2H; 3.63, s, 2H; 3.72, s, 3H; 7.08-7.31, m, 4H.

Step 2: The above ester is hydrolyzed using the procedure described in Example 6, step 2, to give 1,2-bis- [m- (carboxymethyl] ethane as a white solid. MS (HIAD) m / z 297 (M ⁺ -1, 100%). ¹ H NMR (200 MHz, d ₆ -DMSO): d 2.82, s, 2H; 3.56, s, 2H; 7.06-7.06-6.27, m 4H; 12.25; bs, 1H.

Experience 10

In this experiment, the synthesis of 1- [m- (carboxymethyl) phenyl] -2- [m- (carboxyphenyl)] ethane is presented:

Stage 1: Methyl 3- (ethenyl) phenylacetate (Example 8, stage 1) (1.1 g, 6.25 mmol), methyl 3-bromobenzoate (960 mg, 4.46 mmol), palladium acetate (20 mg, 0 , 09 mmol), N, N-dimethylglycine hydrochloride (249 mg, 1.78 mmol) and sodium acetate (731 mg, 8.92 mmol) are dissolved in N-methylpyrrolidinone, degassed with argon and heated to 130 ° C for 5 hours . The reaction mixture was cooled to rt, diluted with EtOAc (100 ml), and the organic phase was washed with water (100 ml), aqueous HCl (1 M, 100 ml) and saturated aqueous NaHCO ₃ (100 ml). The organic extracts were dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo to give trans-1- [m- (3-carboxymethoxymethyl) phenyl] -2- [3- (carbomethoxyphenyl)] ethane as a colorless oil. MS (HIAD) m / z 309 (M ⁺ -1, 100%). ¹ H NMR (200 MHz, CDCl ₃ ): d 3.64, s, 2H; 3.68, s, 3H; 7.16-7.56, 6H; 7.63-7.71, m, 1H; 7.90-7.98, m, 1H; 8.20, m, 1H.

Stage 2: The above compound is hydrogenated in accordance with the method described in Example 9, stage 1, to obtain 1- [m- (carbomethoxymethyl) phenyl] -2- [m- (carbomethoxyphenyl)] ethane as a colorless oil. ¹ H NMR (200 MHz, CDCl ₃ ): d 2.87, m, 4H; 3.56 s, 2H; 3.60, s, 3H; 3.84, s, 3H; 6.95-7.36, 6H; 7.77-7.90, m, 2H.

Stage 3: The ether obtained in Stage 2 is hydrolyzed using the method described in Example 6, stage 2, to obtain 1- [m- (carboxymethyl) phenyl] -2- [m- (carboxyphenyl)] ethane as a white solid . MS (HIAD) m / z 283 (M ⁺ -1, 100%). ¹ H NMR (200 MHz, d ₆ -DMCO): d 2.92, m, 4H; 3.55, s, 2H; 7.02-7.35, m, 4H; 7.36-7.60, m, 2H; 7.71-7.93, m, 2H. ¹³ C NMR (50 MHz, d ₆ -DMSO): d 38.6, 38.7, 40.9, 128.5, 128.8, 130.0, 130.3, 131.0, 131.2, 132.6, 134.8, 136.7, 143.1, 143.8, 169.2, 174.5.

Experience 11

In this experiment, a synthesis of N, N-bis (m-carboxybenzyl) glycine is presented:

Stage 1: m-Cyanobenzyl bromide (2.35 g, 12.0 mmol) is slowly added to a solution of glycine methyl ether hydrochloride (0.63 g, 5.0 mmol), NaHCO ₃ (1.4 g, 17.0 mmol) and NaI (0.37 g, 2.4 mmol) in DMSO (5 ml) and THF (20 ml). The reaction mixture was heated to reflux for 2 hours, cooled to room temperature and diluted with EtOAc (50 ml) and water (40 ml). The organic phase is washed with water (3 × 40 ml), saturated aqueous NaCl (40 ml), dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo to give N, N-bis (m-cyanobenzyl) glycine methyl ether as a colorless oils of sufficient purity for subsequent reactions. Further purification can be carried out by extraction with a dilute aqueous solution of acid, alkalization and extraction with an organic solvent. ¹ H NMR (200 MHz, CDCl ₃ ): d 3.39, s, 2H; 3.71, s, 3H; 3.86, s, 4H; 7.39-7.73, m, 10H.

Stage 2: The above nitrile (1.5 g, 5.02 mmol) is hydrolyzed in accordance with the method described in Example 4, stage 2, to obtain N, N-bis (m-carboxybenzyl) glycine (sulfate salt) as a whitish solid matter. MS (HIAD) m / z 342 (M ⁺ -1, 100%). ¹³ C NMR (50 MHz, d _4- MeOH): d 53.8, 59.0, 130.2, 131.5, 132.7, 132.8, 134.2, 136.1, 169.1, 170.7.

Experience 12

This experiment presents the synthesis of (3R, 4R) -1,3-bis- (p-methoxybenzyl) -4,5-bis (m-phosphonophenyl) imidazolid-2-one:

Step 1: p-Methoxybenzylamine (7.42 g, 54 mmol) in CH ₂ Cl ₂ (100 ml) containing anhydrous MgSO ₄ was treated with m-bromobenzaldehyde (10.0 g, 54 mmol) at 0 ° C. The reaction mixture allowed to stir at room temperature for 16 hours, filtered and concentrated in vacuo to give N-p-methoxybenzylimine m-bromobenzaldehyde as a colorless oil. ¹ H NMR (200 MHz, CDCl ₃ ): d 3.82, s, 3H; 4.78, s, 2H; 6.92, d, J = 7.5 Hz, 2H; 7.16, d, J = 7.5 Hz, 2H; 7.52-7.60, m, 1H; 7.62-7.72, m, 1H; 7.98, m, 1H; 8.29, m, 1H.

Stage 2: 1,2-Dibromoethane (0.5 ml) was added to zinc (1.31 g, 20.0 mmol) in CH ₃ CN (5 ml), and this mixture was heated to reflux for 1 minute. Once the reaction mixture has cooled to rt, TMCCl (1 ml) is added to it, and the reaction mixture is stirred at room temperature for 1 hour. The above imine (6.08 g, 20 mmol) in CH ₃ CN (20 ml) was added in one portion, followed by the addition of TMSl (3.8 ml) over 30 minutes. Then the reaction mixture was stirred for 4 hours at 35-40 ° C. The reaction mixture was quenched with an aqueous solution of NH ₄ OH (6 ml) and a saturated aqueous solution of NH ₄ Cl (14 ml) and filtered. The aqueous phase was separated, and this aqueous phase was extracted with Et ₂ O (50 ml). The combined organic extracts were dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo to give an orange oil. Column chromatography (SiO ₂ 25% Et ₂ O in petroleum ether) gives (1R, 2R) -N, N '- (p-methoxybenzyl) -1,2- (m-bromophenyl) ethane-1,2-diamine in the form of a colorless oil. ¹³ C NMR (50 MHz, CDCl ₃ ): d 50.5, 55.3, 67.4, 113.8, 122.4, 126.7, 129.2, 129.6, 130.3, 130, 7, 132.1, 143.4, 158.7.

Stage 3: N, N'-disuccinimidyl carbonate (160 mg, 0.64 mmol) is added to a solution of the above diamine (260 mg, 0.43 mmol) in CH ₃ CN (10 ml). The reaction mixture is heated to reflux for 2 hours. A subsequent portion of N, N'-disuccinimidyl carbonate (160 mg, 0.64 mmol) was then added, and the reaction mixture was heated to reflux over the next 2 hours. The reaction mixture was concentrated and partitioned between EtOAc (40 ml) and aqueous HCl (1 M, 40 ml). The organic phase was washed with saturated aqueous NaHCO ₃ solution (40 ml), saturated NaCl (40 ml), dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo to give an orange oil. Column chromatography (SiO ₂ , 25% Et ₂ O in petroleum ether ® EtOAc) gives (3R, 4R) -1,3-bis- (p-methoxybenzyl) -4,5-bis (m-bromophenyl) imidazolid -2-one as a white solid. ¹³ C NMR (50 MHz, CDCl ₃ ): d 45.3, 55.2, 65.0, 111.4, 123.1, 125.9, 128.1, 129.8, 130.2, 130, 5, 131.7, 140.7, 159.1, 159.9.

Stage 4: The above urea (200 mg, 0.31 mmol) is treated with diethyl phosphite under the conditions described in Example 3, stage 1, to obtain (3R, 4R) -1,3-bis- (p-methoxybenzyl) -4.5 bis [m- (diethoxyphosphono) phenyl] imidazolid-2-one as a colorless oil. MS (HIAD) m / z 750 (M ⁺ -1, 100%). ³¹ P NMR (81 MHz, proton decoupling, CDCl ₃ ): d +18.4.

Step 5: The above phosphonate (340 mg, 0.45 mmol) was treated with trimethylsilyl bromide under the conditions described in Example 3, step 2, to give (3R, 4R) -1,3-bis- (p-methoxybenzyl) -4.5 bis (m-phosphonophenyl) imidazolid-2-one as an off-white solid. ³¹ P NMR (81 MHz, proton decoupling, CDCl ₃ ): d + 11.5.

Experience 13

In this experiment, the synthesis of 6-amino-4- (4'-pyridyl) -2- (1H) -pyridone is presented:

Stage 1: the interaction of 4,4'-bipyridine with NaNH ₂ in accordance with JOC, 1997, 62, 2774 receive, in addition to the described 2,2'-diamino-4,4'-bipyridine, previously not described 2-amino 4,4'-bipyridine. ¹³ C NMR (50 MHz, d ₆ -DMSO): d 105.0, 109.5, 120.9, 145.4, 148.9, 150.3, 160.5.

Stage 2: The above amino pyridine (1.5 g, 10.5 mmol) is dissolved in acetic anhydride (20 ml) and heated to 60 ° C for 3 hours. The reaction mixture was cooled to room temperature and filtered. The solid was washed with Et ₂ O (2 × 50 ml) and dried in vacuo to give 2- (acetylamino) -4- (4'-pyridyl) pyridine as a light brown solid. ¹ H NMR (200 MHz, d ₆ -DMSO): d 2.12, s, 3H; 7.40-7.58, m, 1H; 7.60-7.83, m, 2H; 8.30-8.58, m, 2H; 8.62-8.88, m, 2H.

Stage 3: The above pyridine (0.9 g, 4.2 mmol) was dissolved in CH ₂ Cl ₂ (50 ml) and treated with m-chloroperbenzoic acid (4.86 g, 60 wt.%) And the reaction mixture was heated to reflux within 16 hours. The reaction mixture was cooled to rt, filtered, and the precipitate was washed with Et ₂ O (2 × 50 ml). The precipitate was added to acetic anhydride (25 ml) and heated to reflux for 4 hours, cooled to room temperature and the precipitate was collected. This precipitate was added to methanol (5 ml), treated with Na ₂ CO ₃ (50 mg) and heated to reflux for 5 hours. The reaction mixture was cooled to rt, filtered and the filtrate was concentrated in vacuo. After trituration with Et ₂ O, 6-amino-4- (4'-pyridyl) -2- (1H) -pyridone is obtained as a yellow solid. MS (CI) m / z 188 (M ⁺ -1, 100%). ¹ H NMR (200 MHz, d ₄ -MeOH): d 5.83, d, J = 1 Hz, 1H, 5.90, d, J = 1 Hz, 1H; 7.67, d, J = 7 Hz, 2H; 8.16, d, J = 7 Hz, 2H.

Experience 14

In this experiment, the activity of some compounds of the present invention in modulating the Fc receptor is displayed.

The interaction between recombinant soluble FcgRIIa and human immunoglobulin in the presence of small amounts of the compounds shown in FIG. 5A and 5B, examined using a BIAcore 2000 biosensor (Pharmacia Biotech, Uppsala, Sweden) at 22 ° C in Hepes buffered saline [NBR: 10 mM Hepes (pH 7.4), 150 mM NaCl, 3.4 mM EDTA, 0.005% surfactant P20 (Pharmacia)]. Monomeric human IgG1, IgG3 and IgE (50 mg / ml) (non-specific binding control) are covalently coupled to the surface of carboxymethylated dextran on a CM-5 sensor chip (BIAcore, Uppsala, Sweden) using an amino group binding protocol (BIAcore, Uppsala, Sweden ) An additional channel is chemically treated using a binding protocol. Recombinant soluble FcgRIIa is used at a concentration of 125 mg / ml, which is equivalent to 50% binding. Recombinant soluble FcgRIIa was preincubated with each of the compounds at room temperature for 30 minutes before being applied to the surface of the sensor chip for 1 minute at a rate of 10 ml / min, followed by a 3-minute dissociation phase. All surfaces are regenerated with 50 mM diethylamine (around pH 11.5), 1 M NaCl between assays of each compound. Then measure the maximum response for each of the interactions. Nonspecific binding responses (IgE channel) are subtracted from the values for binding to IgG1 and IgG3. Measurements are adjusted according to differences in the composition of the buffer between the compounds and the receptor.

Using the sensitivity of surface plasma resonance, measure the degree of interaction of IgG1 (Fig. 6 and 8) and IgG3 (Fig. 7 and 9) with soluble FcgRIIa in the presence of various compounds. Compounds BRI6728, BRI6734, BRI6813, BRI6800, BRI6801, BRI6802, BRI6803, BRI6814, BRI6817, BRI6822, BRI6823 and BRI6824 inhibit the interaction of soluble FcgRIIa with IgG1 (Fig. 6 and 8). At a concentration of 5 mg / ml, compounds BRI6798, BRI6799, BRI6815 and BRI6825 enhance the interaction between soluble FcgRIIa with IgG1 (Fig. 6 and 8). Compounds BRI6728, BRI6734, BRI6813, BRI6800, BRI6801, BRI6802, BRI6803, BRI6814, BRI6816, BRI6817, BRI6822, BRI6823 and BRI6824 inhibit the interaction of soluble FcgRIIa with IgG3 (Fig. 7 and 9). Compounds BRI6727, BRI6798, BRI6815, and BRI6825 enhance the interaction between soluble FcgRIIa with IgG3 at a concentration of about 5 mg / ml and 10 mg / ml.

Experience 15

This experiment presents the synthesis of N- (3’-carboxyphenyl) -2- (carboxybenzene) sulfonamide:

Step 1: Methyl 2- (chlorosulfonyl) benzoate (2.25 g, 8.73 mmol) in methylene chloride (20 ml) was added dropwise to a solution of ethyl 3-aminobenzoate (1.44 g, 8.73 mmol) and triethylamine (1.21 ml, 8.73 mmol) in methylene chloride (10 ml) at 0 ° C. The reaction mixture was allowed to warm to room temperature and stirred overnight. The reaction mixture was washed with water (20 ml), aqueous HCl (1 M, 20 ml) and aqueous NaOH (1 M, 20 ml), dried (MgSO ₄ ), filtered and concentrated in vacuo to give an orange oil. After trituration with ethyl ether, N- (3'-carboethoxyphenyl) -2- (carbomethoxy) benzene sulfonamide is obtained as a white solid. ¹ H NMR (200 MHz, CDCl ₃ ): d 1.31, t, J = 6.0 Hz, 3H; 4.00, s, 3H; 4.29, q, J = 6.0 Hz, 2H; 7.23-7.61, m, 5H; 7.66-7.92, m, 3H; 8.26. br s, 1H.

Stage 2: The above diester (1.0 g, 2.75 mmol) is hydrolyzed using the procedure described in Example 6, stage 2, to obtain N- (3'-carboxyphenyl) -2- (carboxybenzene) sulfonamide as a white solid substances. MS (CI) m / z 320 (M ⁺ -1, 100%). ¹³ C NMR (50 MHz, d ₆ -DMSO): d 168.0, 166.3, 137.3, 135.8, 133.4, 132.6, 131.3, 130.1, 129.0, 128.8, 128.0, 124.5, 123.8 and 120.5.

Experience 16

In this experiment, a synthesis of trans-3,3'-bis- (N-hydroxyamidino) stilbene is presented:

Stage 1: trans-3,3'-Dicyanostilbene obtained from 3-bromobenzonitrile by the method of Example 6, stage 1. MS (CI) m / z 230 (M ⁺ , 100%).

Stage 2: trans-3,3'-Dicyanostilbene (1.5 g, 6.52 mmol), hydroxylamine hydrochloride (3.26 g, 50 mmol) and Na ₂ CO ₃ (3.04 g, 30 mmol) in EtOH (40 ml) and water (15 ml) are heated to reflux for 3 hours. The reaction mixture was cooled to room temperature, and ethanol was removed in vacuo. The remaining solution was extracted with EtOAc (2 × 50 ml), and the combined organic extracts were washed with aqueous HCl (1 M, 2 × 20 ml). The combined aqueous extracts were made basic and extracted with EtOAc (3 × 50 ml). The combined organic extracts were dried (MgSO ₄ ), filtered and concentrated in vacuo to give a colorless solid. MS (CI) m / z 297 (M ⁺ +1, 100%). ¹³ C NMR (50 MHz, d ₆ -DMSO): d 123.3, 124.8, 127.1, 128.6, 133.8, 136.8 and 150.7.

Experience 17

This experiment presents the synthesis of (d, 1) - and meso-2-acetylamino-3- (3- {2- [3- (2-acetylamino-2-carboxyethyl) phenyl] ethyl} phenyl) propionic acid:

Stage 1: 3-Bromobenzaldehyde (23.7 g, 128.2 mmol), N-acetylglycine (10.0 g, 85.5 mmol) and sodium acetate (5.26 g, 64.1 mmol) in acetic anhydride ( 60 ml) are heated to reflux for 1 hour. The reaction mixture was cooled to room temperature and water (100 ml) was added. The resulting suspension was filtered, and the solid was washed with water (2 × 50 ml). The remaining solid was dissolved in methylene chloride (100 ml), dried (MgSO ₄ ), filtered and concentrated in vacuo to give a yellow solid. This solid was suspended in dry MeOH (200 ml) and heated to reflux for 9 hours. The reaction mixture was concentrated in vacuo to give a yellow solid. Recrystallization from EtOAc and petroleum ether gave methyl m-bromo-α-acetamidocinnamate as a yellow solid. MS (CI) m / z 298 (M ⁺ +1 (Br = 79), 100%), 300 (M ⁺ +1 (Br = 81), 100%). ¹³ C NMR (50 MHz, d ₆ -DMCO): d 23.3, 52.8, 122.5, 125.0, 128.06, 130.0, 130.2, 132.2, 132.3, 135.9, 165.4 and 168.8.

Step 2: Trans-Methyl 2-acetylamino-3- (3- {2- [3-trans- (trans-2-acetylamino-2-carbomethoxyethenyl) phenyl] ethenyl} phenyl) prop-2-enoate is prepared from the aforementioned compound by the method Example 6, stage 1. MS (CI) m / z 461 (M ⁺ -1, 100%).

Step 3: The compound from Step 2 (380 mg, 0.82 mmol) and Pd / C (300 mg, 10%) in MeOH (20 ml) was stirred under a hydrogen atmosphere at room temperature for 16 hours. The reaction mixture was filtered and concentrated in vacuo to give (d, 1) - and meso-methyl 2-acetylamino-3- (3- {2- [3- (2-acetylamino-2-carbomethoxyethyl) phenyl] ethyl } -phenyl) -propanoate in the form of a clear viscous oil, which is used without further purification.

Stage 4: The compound from stage 3 (280 mg, 0.60 mmol) is hydrolyzed using the method described in Example 6, stage 2, to obtain (d, 1) - and meso-2-acetylamino-3- (3- { 2- [3- (2-acetylamino-2-carboxyethyl) phenyl] ethyl} phenyl) propionic acid as a clear, viscous oil. MS (CI) m / z 440 (M ⁺ -1, 100%).

Experience 18

In this experiment, the synthesis of (3R, 4R) -4,5-bis (m-carboxyphenyl) imidazolid-2-one is presented:

Step 1: To a solution of (R, R) -1,2-bis- [3- (carbomethoxy) phenyl] ethane-1,2-diol (Example 7, step 1) (1.5 g, 4.54 mmol) in pyridine (10 ml) at 0 ° C. methanesulfonyl chloride (1.01 ml, 13.1 mmol) was added dropwise. The reaction mixture was allowed to warm to room temperature and was stirred overnight. The reaction mixture was diluted with water (30 ml) and 30 ml of methylene chloride, and the aqueous phase was extracted with 2 × 10 ml of methylene chloride. The combined organic extracts are washed with 2 × 20 ml of 1 M aqueous HCl, 20 ml of an aqueous sodium bicarbonate solution, dried over magnesium sulfate, filtered and concentrated in vacuo to give (R, R) -1,2-bis [3] - (carbomethoxy) phenyl] ethane-1,2-diol as a yellow viscous oil.

Stage 2: A solution of the above mesylate (505 mg, 1.0 mmol) and NaN ₃ (150 mg, 2.31 mmol) in 6 ml of DMF is heated to 90 ° C for 17 hours. The reaction mixture was cooled to rt, diluted with 50 ml of diethyl ether and washed with 3 × 50 ml of water. The organic phase was dried over MgSO ₄ , filtered and concentrated in vacuo to give (R, R) -1,2-bis-3- (carbomethoxy) phenyl] -1,2-diazo-ethane as a yellow viscous oil. ¹ H NMR (200 MHz, CDCl ₃ ): d 3.93, s, 3H; 4.73, s, 1H; 7.17-7.39, m, 2H; 7.78-8, 01, m, 2H.

Step 3: The above diazide (611 mg, 1.61 mmol) and Pd on carbon (10%, 50 mg) in methanol are treated with concentrated aqueous HCl (3.86 ml, 3.86 mmol).

The reaction mixture was placed in a hydrogen atmosphere and stirred at room temperature for 30 hours. The reaction mixture was filtered through celite and concentrated to give the (R, R) -1,2-bis- [3- (carbomethoxy) phenyl] -1,2-diamino-ethane hydrochloride salt. MS (CI) m / z 329 (M ⁺ +1 for free amine, 70%), 312 (100%).

Step 4: The above diamine (free base form) (280 mg, 0.85 mmol) in 5 ml of acetonitrile is treated with DMAP (104 mg, 0.85 mmol) with a solution of di-tert-butyl dicarbonate (204 mg, 0 , 94 mmol) in 1 ml of acetonitrile at rt The reaction mixture was stirred for 25 minutes at room temperature and partitioned between 50 ml of ether and 50 ml of 1 M HCl. The organic phase is separated, dried over sodium sulfate, filtered and concentrated in vacuo. Column chromatography yields (3R, 4R) -4,5-bis- (m-carbomethoxyphenyl) imidazolid-2-one as a white solid. MS (HIAD) m / z 355 (M ⁺ +1, 100%). ¹ H NMR (200 MHz, d ₆ -DMSO): d 3.86, s, 6H; 4.57, s, 2H; 7.16, br s, 2H; 7.46-7.61, m, 4H; 7.88-8.00, m, 4H.

Step 5: The above diester (68 mg, 0.19 mmol) is hydrolyzed using the procedure described in Example 6, step 2, to give (3R, 4R) -4,5-bis (m-carboxyphenyl) imidazolid-2-one in the form of a white solid. MS (electrospray) m / z 327 (M ⁺ +1, 100%). ¹³ C NMR (50 MHz, d ₆ -DMSO): d 64.3, 127.4, 129.1, 129.3, 131.1, 131.4, 142.1, 162.5, 167.3.

Experience 19

This experiment presents the synthesis of 3 - ([3 ’- (1’ ’- oxo-2’ ’, 2’ ’, 2’ ’- trifluoroethyl) phenoxy] methyl) phenyl trifluoromethyl ketone:

To a solution of 3 - [(m-bromophenyl) methoxy] bromobenzene (Example 2, step 1) (233 mg, 3.68 mmol) in 6 ml of THF at -78 ° C, tert-butyl lithium (1.6 ml, 1.7 M in pentane, 2.72 mmol). After 20 minutes at this temperature, this solution was added dropwise to a solution of ethyl trifluoroacetate (0.35 ml, 2.94 mmol) in 5 ml of THF at -78 ° C. The reaction mixture was stirred for 16 hours and during this time the reaction mixture reached c.t. The reaction mixture was partitioned between 20 ml of 1 M Hcl and 50 ml of ether. The organic phase is separated, dried over sodium sulfate, filtered and concentrated in vacuo to give 3 - ([3 '- (1''- oxo-2``, 2'',2''- trifluoroethyl) phenoxy] methyl) phenyl trifluoromethyl ketone in colorless oil. MS (CI) m / z 377 (M ⁺ +1, 100%), ¹⁹ F NMR (188 MHz, CDCl ₃ -): d-71.76 and -71.90.

Experience 20

In this experiment, the synthesis of Ac-Phe-Gln-Asn-Gly-Lys-Ser-NH ₂ is presented:

This peptide is synthesized by the synthesis of the peptide on a solid phase. After N-acylation and cleavage of the resin, the title compound is obtained as a white solid. HPLC and MS analyzes confirm the purity and identity of this material.

Experience 21

This experiment presents a synthesis of cyclo- [N-phenylglycine-Gln-Asn- (D) -Asp] -Lus-Ser-NH ₂ :

Step 1: N - [(4S) -3-benzyloxycarbonyl-5-oxo-oxazolidin-4-yl] -acetyl chloride (3.00 g, 10 mmol) in dichloromethane (20 ml) is added dropwise to a solution of tert-butyl- N-phenyl glycinate (2.3 mg, 11 mmol) in pyridine (10 ml) at 0 ° C. The reaction mixture was allowed to warm to room temperature and was stirred overnight. The reaction mixture was diluted with H ₂ O (100 ml) and EtOAc (150 ml). The organic phase is separated and washed successively with citric acid (10%, 2 × 100 ml) and brine (100 ml), dried (MgSO ₄ ), filtered and concentrated in vacuo to give a yellow viscous oil. Column chromatography (SiO ₂ , 20-50% EtOAc in petroleum ether) gives tert-butyl N - [(4S) -3-benzyloxycarbonyl-5-oxo-oxazolidin-4-yl-acetyl] -N-phenyl glycinate as white foam. MS (HIAD) m / z 467 (M ⁺ -1, 100%).

Step 2: An aqueous solution of NaOH (3 ml, 1 M, 3 mmol) is added dropwise to a solution of the above dipeptide (570 mg, 1.22 mmol) in methanol (20 ml) at 0 ° C. The reaction mixture is allowed to warm to room temperature. and monitored by TLC. The reaction mixture was then concentrated and partitioned between Et ₂ O (30 ml) and citric acid (10%, 30 ml) at 0 ° C. The aqueous phase was extracted with Et ₂ O (3 × 30 ml), and the combined organic extracts were dried (MgSO ₄ ), filtered and concentrated in vacuo to give a white solid. Column chromatography (SiO ₂ , 2-5%, MeOH in dichloromethane) gives tert-butyl N - [(2S) -N-benzyloxycarbonyl-aspartyl] -β-N-phenylglycinate as a white solid. MS (HIAD) m / z 456 (M ⁺ +1, 90%).

Step 3: The above compound (1.35 g, 2.96 mol) in MeOH (40 ml) containing palladium on carbon (10%, 500 mg) was placed in a hydrogen atmosphere and stirred at room temperature for 16 hours. The reaction mixture was filtered and concentrated in vacuo to give tert-butyl N - [(2S) -aspartyl]) - β-N-phenyl glycinate as a white solid.

Stage 4: The above compound (890 mg, 2.76 mmol), Fmoc-O-Su, i.e. N- (9-fluoromethoxycarbonyloxy) succinimide (932 mg, 2.76 mmol), Na ₂ CO ₃ (880 mg, 8.29 mmol) in dioxane (15 ml) and H ₂ O (15 ml) are stirred at room temperature in within 16 hours. The reaction mixture was diluted with Et ₂ O (100 ml) and H ₂ O (100 ml). The organic layer was separated and extracted with an aqueous solution of Na ₂ CO ₃ (5%, 3 × 100 ml). The combined aqueous extracts were acidified with 10% aqueous citric acid and extracted with EtOAc (3 × 100 ml). These organic extracts are combined, dried (MgSO ₄ ), filtered and concentrated in vacuo to give tert-butyl N - [(2S) -N-Fmoc-aspartyl]) - β-N-phenyl glycinate as a white solid. ¹³ C NMR (50 MHz, d ₆ -DMSO): d 28.0, 36.7, 47.0, 50.4, 52.4, 67.0, 67.3, 82.3, 119.9, 125.2, 125.3, 127.1, 127.7, 127.8, 128.8, 130.1, 141.2, 142.8, 143.7, 143.9, 156.1, 167, 6, 171.6, 174.5.

Stage 5: Solid-phase amino acid synthesis using the aforementioned protected Fmoc dipeptide, followed by cyclization on the resin and bond cleavage to obtain cyclo- [N-phenylglycine-Gln-Asn- (D) -Asp] -Lus-Ser-NH ₂ .

Experience 22

This experiment presents a synthesis of 2- (2’-phenylethal) -α-N-acetyl-lysinamide and its hydrochloride salt:

Step 1: To a mixture of sodium metal (138 mg, 5.98 mmol) in dry ethanol (16 ml) was added 2-cyano-4- (phenethyl) ethylbutanoate (1.0 g, 4.6 mmol), and this mixture was stirred. at room temperature for 30 minutes Then 4-bromo-but-1-ene (0.6 ml, 6 mmol) was added and the mixture was heated at reflux for 16 hours. The resulting suspension was cooled to room temperature, concentrated under reduced pressure, diluted with ether (100 ml) and NH ₄ Cl (100 ml of a saturated aqueous solution). The aqueous layer was separated and extracted with ether (3 × 50 ml). The organic layers were combined, dried (MgSO ₄ ), filtered and concentrated to give a light brown oil. Column chromatography (silica, eluting with 20% ether / gasoline) gives ethyl 2-cyano-2- (2'-phenethyl) -hex-5-enoate as a clear, colorless oil. ¹ H NMR (200 MHz, CDCl ₃ ): d 1.35 (t, J = 7.0 Hz, 3H), 1.82-2.45 (m, 6H), 2.65 (td, J = 12 , 4 Hz and 7.0 Hz, 1H), 2.90 (td, J = 12.4 Hz and 7.0 Hz, 1H), 4.24 (q, J = 7.0 Hz, 2H), 5 00-5.13 (m, 2H); 5.67-5.76 (m, 1H); 7.15-7.35 (m, 5H).

Step 2: A mixture of the above olefin (0.72 g, 2.65 mmol), LiOH (10.6 ml, 1.0 M, 10.6 mmol) and THF (50 ml) was stirred at room temperature for 18 hours. This reaction mixture was diluted with ether (100 ml) and water (100 ml), and the phases were separated. The aqueous layer was acidified to about pH 2 with 2 M aqueous HCl and transferred to a separatory funnel containing ether (100 ml). The separated aqueous layer was extracted with ether (3 × 50 ml). The organic fractions were combined, dried (MgSO ₄ ), filtered and concentrated under reduced pressure to give 2-cyano-2- (2'-phenethyl) -5-hexenoic acid as a viscous, colorless oil. This material is used in the next reaction without further purification. MS (HIAD) m / z 244 (M ⁺ +1, 55%), 242 (M ⁺ -1, 63%).

Stage 3: Diphenylphosphoryl azide (2.75 ml, 12.8 mmol) and triethylamine (1.75 ml, 12.6 mmol) are added to a solution of the above acid (2.6 g, 10.7 mmol) in toluene (35 ml) . This solution was heated at 100 ° C. for 1 hour, and then tert-butanol (35 ml) was added. This mixture was heated at 100 ° C for the next 2 hours, cooled to room temperature and concentrated under reduced pressure. The resulting yellow oil was diluted with ether (300 ml) and water (300 ml). The organic layer was separated, washed successively with citric acid (100 ml of a 5% aqueous solution), NaHCO ₃ (100 ml of a 5% aqueous solution) and brine (100 ml), dried (MgSO ₄ ), filtered and concentrated to give a yellow oil. Column chromatography (silica, eluting with 2% ethyl acetate / chloroform) gives 2- (N-side-amino) -2- (2 ⁺ phenethyl) -5-hexenonitrile as a colorless oil. MS (HIAD) m / z 316 (M ^{+ +} 1.5%), 313 (M ⁺ -1.2%). ¹³ C NMR (50 MHz, CDCl ₃ ): d 28.4, 30.5, 36.3, 38.9, 54.9, 116.3, 119.7, 126.6, 128.4, 128, 8, 136.3, 140.1, 153.5.

Stage 4: NaOH (9.2 ml, 1.0 M) and H ₂ O ₂ (38 ml 30% (v / v) aqueous solution) are added to a solution of the above nitrile (523 mg, 1.93 mmol) in ethanol ( 20 ml) at 0 ° C. The reaction mixture was stirred at 0 ° C. for 30 minutes and at room temperature for 18 hours. Ethanol was removed under reduced pressure, the residue was diluted with ether (100 ml) and brine (100 ml). The aqueous layer was separated and extracted with ether (4 × 20 ml). The organic layers were combined, dried (MgSO ₄ ), filtered and concentrated to give 2- (N-side-amino) -2- (2'-phenethyl) -5-hexenamide as a colorless sticky foam. This material is used in the next reaction without further purification of R 0.3 (elution with a mixture of 30% ethyl acetate / gasoline). MS (HIAD) m / z 333 M ^{+ +} 1.5%), 233 (100%).

Step 5: Trifluoroacetic acid (2 ml) was added to a solution of the above amide (480 mg, 1.44 mmol) in dichloromethane (5 ml), and this mixture was stirred at room temperature for 35 minutes. This reaction mixture was concentrated to give 2-amino-2- (2'-phenethyl) -5-hexanamide as a red-brown oil. This material is used in the next reaction without further purification. MS (HIAD) m / z 233 (M ⁺ +1, 100%).

Step 6: Acetic anhydride (2.5 ml) was added to a solution of the above amine (335 mg, 1.44 mmol) in pyridine (2.5 ml) and stirred at room temperature for 21 hours. The resulting red-brown reaction mixture was concentrated under reduced pressure. Column chromatography (silica, elution with 80% ethyl acetate / gasoline, R 0.36) affords (N-acetyl amino) -2- (2'-phenethyl) -5-hexenamide as a straw-colored foam. ¹³ C NMR (50 MHz, CDCl ₃ ): d 24.2, 28.3, 30.4, 35.2, 37.8, 64.0, 115.2, 126.1, 128.4, 128, 5, 137.4, 141.1, 169.4, 175.3.

Step 7: To a solution of the above olefin (130 mg, 0.47 mmol) in dry THF (2 ml), 9-BBN (4.6 ml, 0.5 M solution in THF, 2.30 mmol) was added dropwise. The reaction mixture was stirred at ambient temperature for 18 hours. This mixture was cooled to 0 ° C, and water (0.5 ml), NaOAc (5 ml, 5.0 M aqueous solution) and H ₂ O ₂ (5 ml) were successively added. The resulting mixture was stirred at room temperature for 2 hours and diluted with ethyl acetate (30 ml) and brine (30 ml). The aqueous layer was separated and extracted with ethyl acetate (3 × 10 ml). The organic fractions were combined, dried (MgSO ₄ ), filtered and concentrated under reduced pressure to give a pale yellow oil. Column chromatography (silica, elution with 5% MeOH / ethyl acetate, R 0.4) gives 2- (N-acetyl-amino) -2- (2'-phenethyl) -6-hydroxy-hexanamide as a colorless, sticky foam. MS (HIAD) m / z 293 (M ⁺ +1, 35%), 291 (M ⁺ -1, 35%).

Step 8: Triethylamine (0.1 ml, 0.72 mmol) and methanesulfonyl chloride (0.05 ml, 0.65 mmol) are added to a solution of the above alcohol (88 mg, 0.30 mmol) in dichloromethane (2 ml) at 0 ° C. The resulting mixture was stirred at ambient temperature for 19 hours and diluted with ethyl acetate (30 ml) and brine (30 ml). The aqueous layer was separated and extracted with ethyl acetate (3 × 10 ml). The organic layers were combined, dried (MgSO ₄ ), filtered and concentrated under reduced pressure to obtain 2- (N-acetyl-amino) -2- (2'-phenethyl) -6-methanesulfonyloxy-hexanamide as a tan residue. The crude product is used in the next reaction without further purification. MS (HIAD) m / z 371 (M ⁺ +1, 45%), 369 (M ⁺ -1, 5%).

Step 9: A solution of the aforementioned mesylate (110 mg, 0.30 mmol) and sodium azide (54 mg, 0.83 mmol) in dry DMF (2 ml) are heated at a temperature of from 60 ° C to 65 ° C for 19.5 hours. The orange-colored suspension was cooled to room temperature, concentrated and diluted with ethyl acetate (30 ml) and brine (30 ml). The aqueous layer was separated and extracted with ethyl acetate (4 × 10 ml). The organic fractions were combined, dried (MgSO ₄ ), filtered and concentrated to give 2- (N-acetylamino) -2- (2'-phenethyl) -6-azido-hexanamide as a tan oil. This material is used in the next reaction without further purification. ¹ H NMR (200 MHz, CDCl ₃ ): d 1.20-1.78 (m, 6H), 1.92 (s, 3H), 2.20-3.02 (m, 1H), 3.12 -3.30 (m, 2H), 5.57 (s, 1H), 5.90 (s, 1H), 6.72 (s, 1H), 7.04-7.30 (m, 5H).

Step 10: A suspension of the above azide (95 mg, 0.30 mmol) and 10% Pd per C (18.4 mg) in methanol (2 ml) was hydrogenated at room temperature and atmospheric pressure for 21 hours. The black suspension is filtered through a small layer of silica-celite, which is washed with several portions of methanol (approximately 30 ml). After concentrating the filtrate, a light tan oil is obtained. Column chromatography (silica, eluting with 10% triethylamine / methanol, R 0.22) affords 2- (2'-phenylethyl) -α-N-acetyl-lysine amide as a clear, colorless oil. MS (HIAD) m / z 292 (M ⁺ +1, 100%) 290 (M ⁺ -1, 30%). ¹ H NMR (200 MHz, d _4- MeOD): d 1.10-1.60 (m, 4H), 1.73-1.90 (m, 1H), 1.95 (s, 3H), 1 95-2.35 (m, 2H); 2.40-2.80 (m, 5H); 7.10-7.35 (m, 5H).

A small amount of the amine is converted to the corresponding derivative in the form of the hydrochloride salt by adding a 0.5 M aqueous HCl solution to this amine and concentrating the mixture under reduced pressure.

Experience 23

In this experiment, a synthesis of 4,4’-bis- (3 - [(m-carboxyphenoxy) methyl] -2-pyridone) is presented:

Step 1: To a solution of 3-formyl-4-iodo-2-methoxypyridine (prepared according to the method of Fang et al., J. Org. Chem., 1994, 59, 6142) (98 mg, 0.37 mmol) in methanol (4 ml) was added in one portion solid NaBH ₄ at -5 ° C (28 mg, 0.74 mmol). In this case, vigorous evolution of gas is observed and the yellow reaction solution becomes colorless. The reaction was quenched immediately by adding water (2 ml), and methanol was removed under reduced pressure. The resulting residue was diluted with ethyl acetate (20 ml) and water (20 ml). The aqueous layer was separated and extracted with ethyl acetate (3x10 ml). The organic fractions were combined, dried (MgSO ₄ ), filtered and concentrated to give 3- (hydroxymethyl) -4-iodo-2-methoxypyridine as a colorless crystalline solid. This material is used in the next reaction without further purification. R 0.4 (elution with a mixture of 30% ethyl acetate / gasoline). MS (HIAD) m / z 266 (M ⁺ +1, 100%). ¹ H NMR (200 MHz, CDCl ₃ ): d 3.98 (s, 3H), 4.80 (s, 2H), 7.34 (d, J = 4.0 Hz, 1H), 7.70 ( d, J = 4.0 Hz, 1H).

Stage 2: Methanesulfonyl chloride (0.5 ml, 6.4 mmol) is added dropwise to a solution of the above alcohol (373 mg, 1.41 mmol) and triethylamine (0.95 ml, 6.8 mmol) in dichloromethane (9, 4 ml) at 0 ° C. The resulting mixture was stirred at ambient temperature for 15 hours and diluted with ethyl acetate (150 ml) and brine (150 ml). The aqueous layer was separated and extracted with ethyl acetate (3 × 50 ml). The organic fractions were combined, dried (MgSO ₄ ), filtered and concentrated under reduced pressure to give 3- (chloromethyl) -4-iodo-2-methoxypyridine as a light tan crystalline solid. This material is used in the next step without further purification. MS (HIAD) m / z 284 (M ⁺ +1, 100%). ¹ H NMR (200 MHz, CDCl ₃ ): d 3.95 (s, 3H), 4.65 (s, 2H), 7.28 (d, J = 4.0 Hz, 1H), 7.65 ( d, J = 4.0 Hz, 1H).

Stage 3: Sodium salt of methyl 3-hydroxybenzoate (372 mg, 2.14 mmol) is added in one portion to a solution of the above chloride (399 mg, 1.41 mmol) in dry DMF (7 ml). The orange-colored reaction mixture was stirred at room temperature for 18 hours and diluted with ethyl acetate (150 ml) and water (150 ml). The aqueous layer was separated and extracted with ethyl acetate (3 × 30 ml). The organic layers were combined, dried (MgSO ₄ ), filtered and concentrated to give a brown oil. Column chromatography of this oil (silica, elution with 30% ether / gasoline, R 0.35) gives 4-iodo-2-methoxy-3 - {[(m-carbomethoxy) phenoxy] methyl} -pyridine as a colorless oil . MS (HIAD) m / z 400 (M ⁺ +1, 40%). ¹ H NMR (200 MHz, CDCl ₃ ): d 3.92 (s, 3H), 3.96 (s, 3H), 5.20 (s, 2H), 7.15-7.42 (m, 3H ), 7.62-7.80 (m, 3H).

Stage 4: Suspension of the aforementioned iodide (0.5 g, 1.25 mmol), Pd (PPh ₃ ) ₄ (141 mg 0.13 mmol), K ₂ CO ₃ (518 mg, 3.76 mmol), diborpinacone ether ( 159 mg, 0.63 mmol) in DMF (7.6 ml) was heated at 80 ° C, protecting from exposure to light, for 16 hours. The dark brown reaction mixture was cooled to room temperature and diluted with ethyl acetate (150 ml) and water (150 ml). The aqueous layer was separated and extracted with ethyl acetate (3 × 25 ml). The organic layers were combined, dried (MgSO ₄ ), filtered and concentrated under reduced pressure to give a brown oil. Column chromatography of this oil (silica, elution with 50% ethyl acetate / gasoline, R 5.57) gives 4,4'-bis-2-methoxy-3 - {[(m-carbomethoxy) phenoxy] methyl} pyridine in in the form of foam. MS (HIAD) m / z 545 (M ⁺ +1, 100%).

Step 5: A solution of the above dimeric diester (277 mg, 0.51 mmol) in LiOH (10 ml, 1.0 M) and THF (10 ml) was stirred at room temperature for 18 hours. The crude reaction mixture was then diluted with ether (75 ml) and the phases were separated. The aqueous layer was acidified to pH 2 with a 2.0 M aqueous HCl solution and then extracted with ethyl acetate (4 × 50 ml). The organic fractions were combined, dried (MgSO ₄ ), filtered and concentrated to give 4,4'-bis-2-methoxy-3 - {[(m-carboxy) phenoxy] methyl} pyridine as a colorless solid. This material is used in the next step without further purification. MS (HIAD) m / z 517 (M ⁺ +1, 100%), 515 (M ⁺ -1, 100%).

Step 6: Hydrolysis of the aforementioned methoxy-pyridine gives 4,4'-bis- {3 - [(m-carboxyphenoxy) methyl] -2-pyridone}.

Experience 24

This experiment presents the activity of modulating the Fc receptor with a tripeptide and hexapeptide.

The preparation of the peptide.

To obtain an acetylated tripeptide with the GKS sequence and a hexapeptide with the FQNGK-S sequence, the method of solid-phase peptide synthesis (TFPS) is used. See, for example, Merrifield, J. Am. Chem. Soc., 1963, 85, 2419, and Merrifield et al., Anal. Chem., 1966, 38, 1905. These peptides were synthesized using a synergistic peptide synthesizer model 432A. Peptide assembly is based on the chemical properties of Fmoc (Carpino et al., J. Org. Chem. 1972, 37, 3404) using amidated C-terminal resins as starting material. After the assembly of the peptides was completed, an active ester was obtained which reacted with the peptide to form the acetylated N-terminus.

Then, standard operations for breaking the bond using TFA (Fmoc compatible) are applied and the product is purified by reverse phase high performance liquid chromatography (HPLC-RP). (See e.g. Mant, CT and Hodges, RS eds, 1991, "High-Performance Liquid Chromatography of Peptides and Proteins: Separation, Analysis and Confirmation," CRC Press, Boca Raton, Florida). The two mobile phases are 0.1% trifluoroacetic acid (TFA) / 99% H ₂ O and 0.1% TFA / 60% CH ₃ CN / 39.9% H ₂ O. The stationary phase is a preparative Brownlee Column C8 column . The final product was subjected to mass spectral analysis, which confirmed the identity and degree of purity of more than 95% for both peptides.

FcgRIIa binding assays in the presence of hexa or tripeptides

Assays for the interaction between the baculovirus FcgRIIa derivative and the peptide (tripeptide: GKS, hexapeptide: FQNGKS) were performed using a BIAcore 2000 biosensor (Pharmacia Biotech, Uppsala, Sweden) at 22 EC in Hepes buffered saline (NBR: 10 mM Hepes, (pH 7 4), 150 mM NaCl, 3.4 mM EDTA, 0.005% P20 surfactant (Pharmacia). Monomeric human IgG1, IgG3 and IgE (50 mg / ml) are covalently linked to the surface of the CM-5 carboxymethylated dextran (BIAcore, Uppsala , Sweden) using an amino group binding protocol (BIAcore, Uppsala, Sweden). An unbound Ig channel is also chemically processed using rotocol binding: FcgRIIa at a specific concentration (50 mg / ml, concentration providing 50% binding) is mixed with peptides at various concentrations (see FIGS. 10 and 11) for 1 hour at 22 EC before this mixture is applied on the surface of the sensor chip for 1 min at a speed of 20 ml / min, followed by a 3-minute phase of dissociation.After measuring the concentration dependences, all surfaces are regenerated with 50 mM diethylamine (pH 11.5), 1 M NaCl. Then determine the overall response for each concentration of the peptide and plotted against the concentration of the peptide. Non-specific binding responses (IgE channel) are subtracted from IgG1 or IgG3 binding responses.

results

Using the sensitivity of surface plasmon resonance (SPR), the binding of soluble FcgRIIa to IgG1 and IgG3 is determined in the presence of a hexapeptide (FQNGKS) or tripeptide (GKS). In the presence of a hexapeptide, the binding of soluble FcgRIIa to immobilized IgG1 was enhanced four times and 1.6 times upon interaction with IgG3 (Fig. 10). However, the interaction of soluble FcgRIIa with IgG1 or IgG3 in the presence of a tripeptide was inhibited by similar concentrations of this peptide (0-4 mg / ml, Fig. 11).

Experience 25

This experiment presents platelet aggregation inhibition activity for certain compounds of the present invention. In general, the method involves adding a compound to a mixture of platelets and gamma globulin aggregated by heat treatment (HAGG). Out of touch with any of the theories, it is believed that this method reflects the ability of a compound to inhibit the formation of platelet aggregates, as well as its ability to destroy platelet aggregates that formed prior to the addition of this compound.

Platelets express one class of FcgRIIa gamma receptors. After crosslinking of FcgFIIa, platelets undergo various biochemical and cellular changes, which are completed by their aggregation. Using an analysis that is specifically designed to measure platelet aggregation, the ability of the compounds to inhibit platelet activation was measured.

Materials and methods

Platelets are isolated as follows: 30 ml of fresh whole blood is collected in selected tubes with citrate buffer and centrifuged at 1000 rpm for ten minutes. Platelet-rich plasma is separated and centrifuged at 2000 rpm for five minutes in four tubes. Supernatants are removed and platelets are carefully resuspended in 2 ml Tyrodes buffer per tube (137 mM NaCl, 2.7 mM KCl, 0.36 mM NaH ₂ PO ₄ , 0.1% dextrose, 30 mM sodium citrate, 1.0 mM MgCl ₂ · 6H ₂ O, pH 6.5) and again centrifuged at 2000 rpm for five minutes. The supernatants are again removed and the platelets resuspended in 0.5 ml per Hepes tube containing Tyrodes buffer (137 mM NaCl, 2.7 mM KCl, 0.36 mM NaH ₂ PO ₄ , 0.1% dextrose, 5 mM Hepes, 2 mM CaCl ₂ , 1.0 mM MgCl ₂ · 6H ₂ O, pH 7.35). Platelet counts are determined using a hematology analyzer (Coulter) and adjusted to a concentration of approximately 100 × 10 ⁵ platelets / ml using Hepes Tyrodes-containing buffer.

For each aggregation experiment, a mixture containing 50 ml of the Fc receptor agonist, gamma globulin, aggregated by heat treatment ("HAGG", 200 mg / ml) or collagen (2 mg / ml) is incubated with 50 ml of phosphate buffered saline (“PBS”: 3.5 mM NaH ₂ PO ₄ , 150 mM NaCl) or BRI compound (5 mg / ml in PBS) for 60 minutes at room temperature. Then an analysis is performed using a two-cell aggregometer at 37 ° C as follows: glass cuvettes are placed in holders and pre-heated to 37 ° C and 400 ml of platelet suspension are added. After baseline stability has been achieved, 100 ml of the HAGG: PBS, HAGG: BRI or collagen: PBS, collagen: BRI compounds are added to the platelet suspension. Subsequent platelet aggregation is recorded for 15 minutes or until aggregation is complete. Aggregation rate is determined by measuring the gradient and steepness of the aggregation curve.

results

The ability of the compounds (BRI6855, BRI6803, BRI6813, BRI6864, BRI6856, BRI6868, BRI7002) to inhibit HAGG-induced FcgRIIa-dependent aggregation was determined. Platelet aggregation rate, measured as the ratio of transparency increment (y) to time (x), see, for example, FIG. 12 and 13, in the presence of compounds BRI6855, BRI6803, BRI6813, BRI6864 and BRI6856 decreases compared to the speed achieved using the FcgRIIa agonist and gamma globulin aggregated by heat treatment (100%), see Table 1. BRI6868 compounds and BRI7002 do not show the ability to significantly inhibit platelet activation rate. Table 1. Compounds BRI6855 and BRI6803 reduce HAGG-induced platelet aggregation, but do not significantly inhibit platelet aggregation induced by collagen. This indicates that the effects of BRI6855 and BRI6803 are specific for HAGG.

Experience 26

This experiment presents platelet aggregation inhibition activity for certain compounds of the present invention. In general, the method comprises adding a compound to a mixture of platelets and GATO. Out of touch with any of the theories, it is believed that unlike Experience 25, this method only displays the ability of the compound to inhibit platelet aggregation

Materials and methods

To isolate platelets and determine the number of platelets, the experimental method of Experience 25 was used.

In contrast to Test 25, a platelet aggregation assay is performed by adding 50 ml of PBS or BPI compound to a platelet suspension. After approximately one minute, 50 ml of agonist (GATO, collagen or ADP) is added to the platelet suspension. Subsequent platelet aggregation is recorded for 10-15 minutes, or until aggregation is complete. Aggregation rate is determined by measuring the gradient and steepness of the aggregation curve.

results

The ability of the BRI6728 compound to inhibit HAGG-induced FcgRIIa-dependent aggregation was determined. Platelet aggregation rate, measured as the ratio of transparency increment (y) to time (x), see, for example, FIG. 14, in the presence of titratable amounts of the compound, BRI6728 decreases compared to the rate achieved using the FcgRIIa agonist and gamma globulin aggregated by heat treatment (100%), see FIG. 14. The results of platelet aggregation using other compounds are presented in Table 2.

Those skilled in the art will recognize that the preferred embodiments of the present invention are capable of numerous changes and modifications, and that such changes can be made without departing from the scope of the present invention. Thus, it is intended that the appended claims cover all such equivalent changes that meet the spirit and limits of the present invention.

Claims (101)

1. A pharmaceutical composition comprising (a) a compound modulating the activity of an Fc receptor selected from the group comprising an aromatic compound of the formula

heteroaromatic compound of the formula

cyclic compound of the formula

bicyclic compound of the formula

and amino acid derivative of the formula

or their salts, where each of W ¹ and W ² independently represents CO ₂ R ¹⁵ , C (= NH) NH (OH), SO ₃ R ¹⁵ , C (= NH) NH ₂ , OPO (OR ¹⁵ ) ₂ , C (= O ) CF ₃ or PO (OR ¹⁵ ) ₂ ; each of Ar ¹ , Ar ² , Ar ⁴ and Ar ⁵ independently is C ₆ -C ₂₀ aryl or C ₁ -C ₂₀ heteroaryl; Ar ³ is C ₁ -C ₂₀ heteroaryl; each of X ¹ , X ² , X ³ , X ⁴ , X ⁵ , X ⁶ , X ⁷ and X ⁸ independently represents methylene, O, S or NR ¹⁶ ; each of R ¹ and R ² independently represents a bond, C ₁ -C ₆ alkylene or halogenated C ₁ -C ₆ alkylene; each of R ³ and R ⁴ independently represents halogen, —Z ¹ or C ₁ -C ₆ alkyl; each of X ⁹ , Y ¹ and Z ¹ independently represents OR ¹⁷ , SR ¹⁷ or NR ¹⁷ R ¹⁸ ; each of R ⁵ and R ⁶ independently represents an amino acid residue of a side chain or a group of the formula —R ¹⁹ —W ³ ; each of R ⁸ , R ⁹ and R ¹¹ independently represents an amino acid residue of the side chain, provided that R ^{11 is} not H or CH ₃ ; R ⁷ represents OR ²⁰ , NR ²¹ R ^22, or from about 1 to 10 amino acids; R ¹⁰ represents C ₁ -C ₆ alkylene; R ¹² represents C ₁ -C ₆ alkyl or C ₆ -C ₂₀ aralkyl; W ³ represents C (= O) X ¹⁰ ; X ¹⁰ represents OR ²³ or NR ²⁴ R ²⁵ each of R ¹³ , R ¹⁵ , R ¹⁷ , R ¹⁸ , R ²⁰ , R ²¹ , R ²³ and R ²⁴ independently represents hydrogen or C ₁ -C ₆ alkyl; R ¹⁶ represents H, C ₆ -C ₂₀ aryl or an amide protecting group; R ¹⁹ represents C ₁ -C ₆ alkylene; each of R ²² and R ²⁵ independently represents H, C ₁ -C ₆ alkyl or an amide protecting group; R ¹⁴ represents H, C ₁ -C ₆ alkyl or an amine protecting group; L represents a linking group comprising from 1 to 20 atoms; and each of m and n independently represents an integer from 0 to 2, and (b) a pharmaceutically acceptable carrier. 2. The composition according to claim 1, where the specified compound has the formula

3. The composition according to claim 2, where the specified compound has the formula

4. The composition according to claim 3, where m and n = 0. 5. The composition according to claim 4, where W ¹ and W ² represent CO ₂ N. 6. The composition according to claim 5, where R ¹ and R ² are a bond. 7. The composition of claim 6, wherein L ¹ is -CH ₂ CH _2. 8. The composition according to claim 6, where L ¹ represents-CH ₂ O-. 9. The composition according to claim 6, where L ¹ represents-CH = CHS (= O) -. 10. The composition of claim 6, wherein L ¹ is —CH ₂ CH ₂ CH (OH) -. 11. The composition according to claim 6, where L ¹ represents —CH = CH—. 12. The composition according to claim 6, where L ¹ represents —CH (OH) CH (OH) -. 13. The composition of claim 12, wherein the stereochemical arrangement of the hydroxyl groups is (S, S). 14. The composition according to claim 6, where L ¹ represents —CH ₂ N (R ²⁶ ) CH ₂ -, where R ²⁶ represents H, C ₁ -C ₆ alkyl or an amine protecting group. 15. The composition of claim 14, wherein R ²⁶ is —CH ₂ CO ₂ N. 16. The composition according to claim 6, where L ¹ represents a group of the formula

17. The composition according to claim 5, where R ¹ and R ² represent —CH ₂ . 18. The composition according to 17, where L ¹ represents ethylene. 19. The composition of claim 17, wherein L ¹ is —CH = CH—. 20. The composition according to claim 5, where R ¹ represents methylene, R ² represents a bond and L ¹ represents ethylene. 21. The composition according to claim 4, where W ¹ and W ² are PO (OR ¹⁵ ) ₂ and R ¹ and R ² are a bond. 22. The composition according to item 21, where L ¹ represents ethylene. 23. The composition of claim 22, wherein R ¹⁵ is ethyl. 24. The composition according to item 22, where R ¹⁵ represents N. 25. The composition according to item 21, where L ¹ represents a group of the formula

where each of R ²⁷ and R ²⁸ independently represents H, C ₁ -C ₆ alkyl, C ₆ -C ₁₀ aralkyl or a protective group. 26. The composition according A.25, where each of R ²⁷ and R ²⁸ independently represents 4-methoxybenzyl or N. 27. The composition according to claim 6, where L ¹ represents a group of the formula

where each of R ²⁷ and R ²⁸ independently represents H, C ₁ -C ₆ alkyl, C ₆ -C ₁₀ aralkyl or a protective group. 28. The composition according to item 27, where each of R ²⁷ and R ²⁸ independently represents 4-methoxybenzyl or N. 29. The composition according to claim 4, where L ¹ represents —CH = CH—, W ¹ and W ² represent C (NH NH) NH (OH) and R ¹ and R ² represent a bond. 30. The composition according to claim 4, where L ¹ represents —CH ₂ O—, W ¹ and W ² represent C (= O) CF ₃ and R ¹ and R ² represent a bond. 31. The composition according to claim 4, where L ¹ represents —CH ₂ CH ₂ -, R ₁ and W ₁ together form - (CH ₂ ) _a CH (NHR ²⁹ ) CO ₂ H, where a = 0, 1, 2 and R ²⁹ represents H, C ₁ -C ₆ alkyl, or an amine protecting group. 32. The composition according to p, where R ² and W ² together form - (CH ₂ ) _b CH (NHR ³⁰ ) CO ₂ H, where b = 0, 1, 2 and R ³⁰ represents H, C ₁ -C ₆ alkyl, or an amine protecting group. 33. The composition of claim 32, wherein a and b = 1 and R ²⁹ and R ³⁰ are —C (═O) CH ₃ . 34. The composition according to claim 2, where the specified compound has the formula

35. The composition according to claim 1, where the specified compound has the formula

36. The composition of claim 35, wherein said compound has the formula

37. The composition of claim 36, wherein Y ¹ is —NH ₂ . 38. The composition according to clause 37, where m and n = 0. 39. The composition according to claim 1, where the specified compound has the formula

where X ¹ , X ² , X ³ , X ⁴ represent NR ¹⁶ . 40. The composition according to § 39, where the specified compound has the formula

41. The composition according to claim 1, where the specified compound has the formula

42. The composition according to claim 1, where the specified compound has the formula

43. The composition according to § 42, where R ¹¹ represents the remainder of the side chain of lysine, R ¹² represents 2'-phenylethyl and R ¹⁴ represents-C (= O) CH ₃ . 44. A method of inhibiting the binding of an Fc receptor to an immunoglobulin of a patient, comprising administering to said patient a pharmaceutically effective amount of a compound selected from the group comprising an aromatic compound of the formula

heteroaromatic compound of the formula

cyclic compound of the formula

bicyclic compound of the formula

and amino acid derivative of the formula

or their salts, where each of W ¹ and W ² independently represents CO ₂ R ¹⁵ , C (= NH) NH (OH), SO ₃ R ¹⁵ , C (= NH) NH ₂ , OPO (OR ¹⁵ ) ₂ , C (= O ) CF ₃ or PO (OR ¹⁵ ) ₂ ; each of Ar ¹ , Ar ² , Ar ⁴ and Ar ⁵ independently is C ₆ -C ₂₀ aryl or C ₁ -C ₂₀ heteroaryl; Ar ³ represents C ₁ -C ₂₀ heteroaryl; each of X ¹ , X ² , X ³ , X ⁴ , X ⁵ , X ⁶ , X ⁷ and X ⁸ independently represents methylene, O, S or NR ¹⁶ ; each of R ¹ and R ² independently represents a bond, C ₁ -C ₆ alkylene or halogenated C ₁ -C ₆ alkylene; each of R ³ and R ⁴ independently represents halogen, —Z ¹ or C ₁ -C ₆ alkyl; each of X ⁹ , Y ¹ and Z ¹ independently represents OR ¹⁷ , SR ¹⁷ or NR ¹⁷ R ¹⁸ ; each of R ⁵ and R ⁶ independently represents an amino acid residue of a side chain or a group of the formula —R ¹⁹ —W ³ ; each of R ⁸ , R ⁹ and R ¹¹ independently represents an amino acid residue of the side chain, provided that R ^{11 is} not H or CH ₃ ; R ⁷ represents OR ²⁰ , NR ²¹ R ^22, or from about 1 to 10 amino acids; R ¹⁰ represents C ₁ -C ₆ alkylene; R ¹² represents C ₁ -C ₆ alkyl or C ₆ -C ₂₀ aralkyl; W ³ represents C (= O) X ¹⁰ ; X ¹⁰ represents OR ²³ or NR ²⁴ R ²⁵ ; each of R ¹³ , R ¹⁵ , R ¹⁷ , R ¹⁸ , R ²⁰ , R ²¹ , R ²³ and R ²⁴ independently represents hydrogen or C ₁ -C ₆ alkyl; R ¹⁶ represents H, C ₆ -C ₂₀ aryl or an amide protecting group; R ¹⁹ represents C ₁ -C ₆ alkylene; each of R ²² and R ²⁵ independently represents H, C ₁ -C ₆ alkyl or an amide protecting group; R ¹⁴ represents H, C ₁ -C ₆ alkyl or an amine protecting group; L represents a linking group comprising from 1 to 20 atoms; each m and n independently represents an integer from 0 to 2. 45. The method according to item 44, wherein said Fc receptor is selected from the group including FcaR, FcgR, FcgR and mixtures thereof. 46. The method of claim 45, wherein said Fc receptor is selected from the group consisting of FcgRIIa, FcgRIIb, FcgRIIc, and mixtures thereof. 47. The method according to item 44, in which IgG-mediated tissue damage in the specified patient is reduced. 48. The method according to item 44, wherein said method reduces inflammation in the specified patient. 49. The method of claim 44, wherein said method is used to treat an autoimmune disease. 50. The method according to item 44, which is used to treat a disease in which antibody aggregates are produced or in which immune complexes are produced as a result of the interaction of the antibody with an internal or external antigen. 51. The method of claim 50, wherein said disease is selected from the group comprising immune complex diseases, autoimmune diseases, infectious diseases, and vasculitis. 52. The method of claim 51, wherein said autoimmune disease is selected from the group consisting of rheumatoid arthritis, systemic lupus erythematosis, immune thrombocytopenia, neutropenia, and hemolytic anemia. 53. The method according to § 51, wherein said vasculitis is selected from the group comprising polyarthritis thickening and systemic vasculitis. 54. The method according to item 44, which is used to treat xenograft rejection. 55. The method according to 51, wherein said infectious diseases are selected from the group comprising hemorrhagic fever caused by Dengue virus and measles viral infection. 56. The method according to item 44, wherein the IgE-mediated response in the specified patient is reduced. 57. The method according to item 44, wherein said compound has the formula

58. The method of claim 57, wherein said compound has the formula

59. The method according to § 58, wherein m and n = 0. 60. The method according to § 59, in which W ¹ and W ² represent CO ₂ N. 61. The method of claim 60, wherein R ¹ and R ² are a bond. 62. The method of claim 61, wherein L ¹ is —CH ₂ CH ₂ . 63. The method of claim 61, wherein L ¹ is —CH ₂ O—. 64. The method of claim 61, wherein L ¹ is —CH = CHC (= O) -. 65. The method of claim 61, wherein L ¹ is —CH ₂ CH ₂ CH (OH) -. 66. The method of claim 61, wherein L ¹ is —CH = CH—. 67. The method of claim 61, wherein L ¹ is —CH (OH) CH (OH) -. 68. The method of claim 67, wherein the stereochemical arrangement of the hydroxyl groups is (S, S). 69. The method of claim 61, wherein L ¹ is —CH ₂ N (R ²⁶ ) CH ₂ -, wherein R ²⁶ is H, C ₁ -C ₆ alkyl, or an amine protecting group. 70. The method according to p, in which R ²⁶ represents-CH ₂ CO ₂ N. 71. The method of claim 61, wherein L ¹ is a group of the formula

72. The method of claim 60, wherein R ¹ and R ² are —CH ₂ . 73. The method of claim 72, wherein L ¹ is ethylene. 74. The method of claim 72, wherein L ¹ is —CH = CH—. 75. The method of claim 60, wherein R ¹ is methylene, R ² is a bond, and L ¹ is ethylene. 76. The method according to § 59, wherein W ¹ and W ² are PO (OR ¹⁵ ) ₂ and R ¹ and R ² are a bond. 77. The method according to p, in which L ¹ represents ethylene. 78. The method according to p, in which R ¹⁵ represents ethyl. 79. The method according to p, in which R ¹⁵ represents N. 80. The method according to p, in which L ¹ represents a group of the formula

where each of R ²⁷ and R ²⁸ independently represents H, C ₁ -C ₆ alkyl, C ₆ -C ₁₀ aralkyl or a protective group. 81. The method of claim 80, wherein each of R ²⁷ and R ²⁸ independently is 4-methoxybenzyl or N. 82. The method of claim 61, wherein L ¹ is a group of the formula

where each of R ²⁷ and R ²⁸ independently represents H, C ₁ -C ₆ alkyl, C ₆ -C ₁₀ aralkyl or a protective group. 83. The method of claim 82, wherein each of R ²⁷ and R ²⁸ independently is 4-methoxybenzyl or N. 84. The method of claim 59, wherein L ¹ is —CH = CH—, W ¹ and W ² are C (= NH) NH (OH), and R ₁ and R ₂ are a bond. 85. The method of claim 59, wherein L ¹ is —CH ₂ O—, W ¹ and W ² are C (═O) CF ₃ and R ¹ and R ² are a bond. 86. The method according to § 59, wherein L ¹ is —CH ₂ CH ₂ -, R ₁ and W ₁ together form - (CH ₂ ) _a CH (NHR ²⁹ ) CO ₂ H, where a = 0, 1, 2 and R ²⁹ represents H, C ₁ -C ₆ alkyl, or an amine protecting group. 87. The method according to p, in which R ² and W ² together form - (CH2) _b CH (NHR ³⁰ ) CO ₂ H, where b = 0, 1, 2 and R ³⁰ represents H, C ₁ -C ₆ alkyl, or an amine protecting group. 88. The method of claim 87, wherein a and b = 1 and R ²⁹ and R ³⁰ are —C (═O) CH ₃ . 89. The method of claim 87, wherein said compound has the formula

90. The method according to item 44, wherein said compound has the formula

91. The method of claim 90, wherein said compound has the formula

92. The method of claim 91, wherein Y ¹ is —NH ₂ . 93. The method according to paragraph 92, wherein m and n = 0. 94. The method according to item 44, wherein said compound has the formula

where X ¹ , X ² , X ³ , X ⁴ represent NR ¹⁶ . 95. The method according to clause 94, wherein said compound has the formula

96. The method according to item 44, wherein said compound has the formula

97. The method according to item 44, wherein said compound has the formula

98. The method of claim 97, wherein R ¹¹ is a lysine side chain residue, R ¹² is 2'-phenylethyl, and R ¹⁴ is —C (= O) CH ₃ . Priority on points: 09/11/1998 according to claims 1-9, 11, 17-24, 44-56, 57-64, 66, 72-79; 04/30/1999 according to claims 1-15, 17-32, 35-38, 44-70, 72-87, 90-93; 08/11/1999 according to claims 1-98.

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