WO2017189569A1

WO2017189569A1 - Multimediator dpp4 and fap inhibitors, and uses related thereto

Info

Publication number: WO2017189569A1
Application number: PCT/US2017/029407
Authority: WO
Inventors: William W. Bachovchin; Hung-Sen Lai; Wengen Wu
Original assignee: Trustees Of Tufts College
Priority date: 2016-04-25
Filing date: 2017-04-25
Publication date: 2017-11-02

Abstract

Disclosed is a pharmaceutical preparation comprising a multimediator inhibitor agent that both inhibits DPP4 enzymatic activity with a Ki of 100 nM or less and inhibits Fibroblast Activation Protein (FAP) enzymatic activity with a Ki of 100 nM or less, which multimediator is orally active and is formulated for oral administration at a dosing frequency of 24 hours or greater.

Description

Muiti mediator DPP4 and FAP Inhibitors, and Uses Related Thereta

Cross-Reference

This a pplication claims the benefit of U .S. Provisiona l Application No. 62/32.7., 240, filed April 25, 2016, the entirety of which is hereby incorporated by reference.

Background

Dipeptidyi peptidase-4 (DPP-4) inhibitors are a relatively new class of oral diabetes drugs. Also known as gliptins, they are usually prescribed for people with type 2 diabetes who have not responded well to drugs such as metformin and suiphonyiureas.

DPP-4 inhibitors may help with weight loss as well as decreasing blood glucose levels.

In recent years, the dipeptidyi peptidase-4 (DPP-4) enzyme family has created intense pharmaceutical interest. DPP-4 inhibitors have proven successful as a therapy for the growing type 2 diabetes epidemic and have potential to treat other diseases. A large number of recently developed DPP-4 inhibitors are in various phases of clinical development, with several gliptin class inhibitors already in clinical use. The unique distribution of Fibroblast Activation Protein (FAP), the closest relative of DPP-4, has led to numerous investigations of it as a target and marker for epithelial cancers. The roles of the newer members, DPP-8 and DPP-9, are yet to be fully characterized, but early evidence suggests possible roles in various aspects of cell biology and disease.

DPP-4 and FAP belongs to the prolyl peptidase family, which comprises serine proteases that typically cleave peptide substrates after a proline residue. This family has been implicated in several diseases, including diabetes, cancer, and mood disorders, and also includes DPP-7, DPP-8, DPP-9, prolyl oiigopeptidase, acylpeptide hydrolase, and prolyl carboxypeptidase. These proteases differ in structure at the N-terminus, but each has a C- terminai αβ-hydrolase domain that contains the catalytic Ser, Asp, and His residues. FAP, like its most closely related family member, DPP-4, is a type Π transmembrane protein; both have a short cytoplasmic tail, a transmembrane domain, and a beta-propeller domain containing several sites of N-linked glycosylation. Crystallographic data for FAP and DPP-4 show that the beta-propeller has important substrate binding sites and suggest that this domain precludes access of large substrates to the αβ-hydroiase domain. DPP-4 regulates biological processes by cleaving regulatory peptides of less than about 10 kDa, including glucagon-like peptide-1, glucose-dependent insuiinotropic peptide, and stromal-derived factor-1. DPP-4 cleaves these peptides via a well characterized dipeptidyi peptidase activity that removes P2-Prol or P2-Aial dipeptides, (P2 represents any amino acid) from the N- terminus of the substrate. DPP-4 similarly cleaves P2-Prol-based synthetic peptides and does so with high catalytic efficiency and broad specificity. In contrast, endogenous peptide substrates of FAP have not previously been known, and the activity of the protease against synthetic substrates has, until this invention, remained pooriy characterized. FAP also cleaves proteins such as gelatinand 2-antiplasmin, suggesting that the two proteases cleave distinct substrates.

There are a number of favorable circumstances that have contributed to the therapeutic approaches of targeting DPP-4 and FAP. Specific targeting of DPP-4 and FAP is made easier by the small size of this enzyme family and several structural differences at their active sites. Additionally, observations over the past decade that the DPP-4 and FAP gene knockout mice are healthy suggests that selective inhibition of each of these proteases would be safe, and this is reflected in the excellent safety profiles of the DPP-4-selective gliptins. These proteins also have interesting extra-enzymatic activities that are expected to be retained in the presence of protease inhibition. This feature also points to a low likelihood of off-target effects. Thus, an overall understanding of DPP-4 and FAP structure- function relationships, distribution, and enzymatic and extra-enzymatic biological roles provides an insight into their therapeutic usefulness as disease targets.

Moreover, given the close substrate specificity relationship between DPP-4 with that of DPP-8 and DPP-9, in developing a single agent capable of selectively and potently DPP-4 and FAP, there is a need for that agent to be a substantially weaker inhibitor of DPP-8 and DPP-9.

Summary of the Invention

One aspect of the invention relates to a pharmaceutical preparation comprising a multimediator inhibitor agent that both inhibits DPP4 enzymatic activity with a Ki of 100 nM or less and inhibits Fibroblast Activation Protein (FAP) enzymatic activity with a Ki of 100 nM or less, which multimediator is orally active and is formulated for oral administration at a dosing frequency of 24 hours or greater.

In certain embodiments, the multimediator inhibits DPP4 enzymatic activity with a Ki of 10 nM or less, for example, less than 1,0 nM, 0.1 nM, 0.01 nM or even 0.001 nM. In certain embodiments, the multimediator inhibits FAP enzymatic activity with a Ki of 10 nM or less, for example, less than 1.0 nM, 0.1 nM, 0,01 nM or even 0.001 nM. In certain embodiments, the multimediator inhibits DPP4 enzymatic activity with a IC50 of 100 nM or less, for example less than 10 nM, 1.0 nM, 0.1 nM, 0.01 nM or even 0.001 nM. In certain embodiments, the multimediator inhibits FAP enzymatic activity with an IC50 of 100 nM or less, for example, less than 10 nM, 1.0 nM, 0.1 nM, 0.01 nM or even 0.001 nM.

In certain embodiments, the multimediator inhibits DPP4 enzymatic activity with a K_0ff rate less than lxl0⁴/sec (for example, less than 5 x lO ⁵ or even 1 x lO ⁵), and inhibits FAP enzymatic activity with a K_0ff rate less than lxl0^"4/sec (for example, less 5 x IfJ⁵ or even 1 x 10- ⁵).

In certain embodiments, the multimediator has a Ki for inhibition of DPP-8 and DPP-9 that is 1 microM or greater, for example, greater than even 1 micro , 10 microM or 100 microM. In certain embodiments, the multimediator has an IC50 for inhibition of DPP-8 and DPP-9 that is, for example, greater than even 1 microM, 10 microM or 100 microM.

In certain embodiments, the multimediator is formulated in a single oral dosage form that delivers an effective amount of the multimediator at a drug dosing frequency of 24 hours or greater, for example, at a drug dosing frequency of 48, 72, 96 hours or even once weekly.

In certain embodiments, the multimediator is formulated in a single oral dosage form containing less than 500 mg (i.e., 0.5 mg-500 mg) of the multimediator, for example, less than 250 mg, 100 mg, 50 mg, or even 10 mg.

In certain embodiments, the pharmaceutical preparation of multimediator also includes one or more additional active ingredients selected from the group consisting of antidiabetic agents, anti-hypergiycemic agents, hypolipidemic/lipid lowering agents, anti- obesity agents, anti-hypertensive agents and appetite suppressants.

In an exemplary embodiment, the multimediator is represented in the general formula:

wherein

R¹ represents a naturally or non-naturally occurring amino acid side chain;

R² represents a moiety including two or more overlapping p-orbitals with delocaiized electrons, which forms a conjugated system with the nitrogen to which R² is attached such that the nitrogen is a tertiary amine which maintains an electron lone pair under conditions in which the multimediator interacts with DPP4;

R⁵ represents a hydrogen, a lower alkyl, or a -lower alkyl-halogen (such as to form a halo-alkyi ketone, i.e., -CH3-CI);

R⁷ represents, for each occurrence, hydrogen, or a substituted or unsubstituted lower alkyl, lower aikenyl, aryl, aralkyl, cycioalkyl, cycloalkenyl, or heterocycie; R⁵⁰ represents O or S;

R⁵¹ represents N₃, SH₂, NH₂, N0₂ or OR⁷;

R⁵² represents hydrogen, a lower alkyl, amine, OR⁷, or a pharmaceutically acceptable salt, or R⁵¹ and R⁵² taken together with the phosphorous atom to which they are attached complete a heterocyclic ring having from 5 to 8 atoms in the ring structure;

Z represents a 4-12 membered heterocycle, which may monocyclic, bicyciic or tricyclic and may include one or more substituents R³ described below;

W represents a functional group which reacts with an active site residue of DPP4 and FAP, as for example,— CN,— -CH^NR⁵,

o o R⁵⁰ o

— S— X¹ , —-P— X¹ ,— P— R⁵² , — & , or— C— R⁵

j "~-γ2

X represents O or S;

X^I represents a halogen; and

Yi and Y₂ can independently or together be OH, or a group capable of being hydrolyzed to a hydroxy! group, including cyclic derivatives where Yi and Y₂ are connected via a ring having from 5 to 8 atoms in the ring structure (such as pinacol or the like).

The muitimediator inhibitor of the present invention can be used for a variety of different disorders, particularly those in which elongation of the serum half-life (lives) or one or more substrates for DPP4 and FAP provides therapeutic benefit.

Such methods can be practiced by administering the muitimediator alone (i.e., as a monotherapy) or in combination one or more other active agents selected from the group consisting of other antidiabetic substances, active substances that lower the blood sugar level, active substances that lower the lipid level in the blood, active substances that raise the HDL level in the blood, active substances that lower blood pressure, active substances that are indicated in the treatment of atherosclerosis or obesity, and/or active substances which are indicated in the treatment or prevention of major CV events and antiplatelet agents and/or anticoagulants.

Brief Description of the Figures

Figures 1 arid 2 demonstrate that ARI-5057, and FAP specific inhibitor (relative to DPP-4) reduces glucose excursion even further in an OGTT when co-administered with sitagiiptin in chow-fed mice at 16 weeks of age. ARI-5057 and sitagiiptin were given P.O. at 20mg/kg 45 minutes before 50% glucose challenge at 2.5g/kg. (N=6-9) per group) . ***P<.QQ1 versus vehicle by ANOVA. This indicates that substrates of FAP, which are not also substrates of DPP-4, play a role in glucose sensitivity.

Figure 3 is a table illustrating several different substrates for FAP cleavage (but not DPP-4 cleavage), indicating the site of FAP cleavage and the potential physiological roles for those proteins/peptides.

Description of Exemplary Embodiments

Overview

The present invention is directed to compositions-of-matter, pharmaceutical preparations, uses in manufacturing medicaments, and methods-of-treatments relating multimediator inhibitor agents that both inhibits DPP4 enzymatic activity with a Ki of 10 nM or less and inhibits FAP enzymatic activity with a Ki of 10 nM or less, which multimediator is orally active and is formulated for oral administration at a dosing frequency of 24 hours or greater.

wherein

R¹ represents a naturally or non-naturally occurring amino acid side chain;

R² represents a moiety including two or more overlapping p-orbitals with deiocalized electrons, which forms a conjugated system with the nitrogen to which R² is attached such that the nitrogen is a tertiary amine which maintains an electron lone pair under conditions in which the multimediator interacts with DPP4;

R⁵ represents a hydrogen, a lower alkyl, or a -lower alkyl-haiogen (such as to form a haio-alkyl ketone, i.e., -CH3-CI);

R⁷ represents, for each occurrence, hydrogen, or a substituted or unsubstitufed lower alkyl, lower alkenyl, aryi, aralkyi, cycloalkyi, cycloalkenyi, or heterocycle; R⁵⁰ represents O or S;

R⁵¹ represents N₃, SH₂, NH₂, N0₂ or OR⁷;

Z represents a 4-12 membered heterocycle, which may monocyclic, bicyclic or tricyclic and may include one or more substituents R³ described below;

W represents a functional group which reacts with an active site residue of DPP4 and FAP, as for example,— CN,— -CH=N R⁵,

o o R⁵⁰ o

— S— X¹ , —-P— X¹ , P R⁵² , — & , or — C— R⁵

j "~-γ2

X represents O or S;

X^I represents a halogen; and

As used herein, the definition of each expression, e.g. alkyl, m, n, etc., when it occurs more than once in any structure, is intended to be independent of its definition elsewhere in the same structure.

The pharmaceutically acceptable salts of the subject compounds include the conventional nontoxic salts or quaternary ammonium salts of the compounds, e.g., from non-toxic organic or inorganic acids. For example, such conventional nontoxic salts include those derived from inorganic acids such as hydrochloride, hydrobromic, sulfuric, sulfamic, phosphoric, nitric, and the like; and the salts prepared from organic acids such as acetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, palmitic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicyclic, sulfaniiic, 2-acetoxybenzoic, fumaric, toluenesulfonic, mefhanesulfonic, ethane disulfonic, oxalic, isothionic, and the like.

The pharmaceutically acceptable salts of the present invention can be synthesized from the subject compound which contain a basic or acid moiety by conventional chemical methods. Generally, the salts are prepared by reacting the free base or acid with stoichiometric amounts or with an excess of the desired salt-forming inorganic or organic acid or base in a suitable solvent. The pharmaceutically acceptable salts of the acids of the subject compounds are also readily prepared by conventional procedures such as treating an acid of Formula I with an appropriate amount of a base such as an al kali or alkaline earth methyl hydroxide (e.g. sodium, potassium, lithium, calcium or magnesium) or an organic base such as an amine, piperidine, pyrrolidine, benzyiamine and the like, or a quaternary ammonium hydroxide such as tetramethylammonium hydroxide and the like.

Contemplated equivalents of the compounds described above include compounds which otherwise correspond thereto, and which have the same general properties thereof (e.g. the ability to inhibit proteolysis of the natural substrates of DPP4 or FAP - such as GLP-1 and FGF21, wherein one or more simple variations of substituents are made which do not adversely affect the efficacy of the compound in use in the contemplated method. In general, the compounds of the present invention may be prepared by the methods illustrated in the general reaction schemes as, for example, described below, or by modifications thereof, using readily available starting materials, reagents and conventional synthesis procedures. In these reactions, it is also possible to make use of variants which are in themselves known, but are not mentioned here.

Also deemed as equivalents are any compounds which can be hydrolyticaliy converted into any of the aforementioned compounds including boronic acid esters and halides, and carbonyl equivalents including acetals, hemiacetals, ketals, and hemiketals.

Definitions

For convenience, before further description of the present invention, certain terms employed in the specification, examples, and appended claims are collected here.

The term "alkyl" refers to the radical of saturated aliphatic groups, including straight-chain alkyl groups, branched-chain alkyl groups, cycloalkyl (alicyclic) groups, alkyl substituted cycioaikyl groups, and cycloalkyl substituted alkyl groups. In certain embodiments, a straight chain or branched chain alkyl has 30 or fewer carbon atoms in its backbone (e.g., Ci -Cso for straight chain, C3-C30 for branched chain), for example 20 or fewer. Likewise, certain cycloalkyls have from 3-10 carbon atoms in their ring structure, for example, 5, 6 or 7 carbons in the ring structure. "Alkyl" (or "lower alkyl") as used throughout the specification and claims is intended to include both "unsubstituted alkyis" and "substituted alkyls".

As used herein, the term "substituted" is contemplated to include all permissible substituents of organic compounds. Such substituents can include, for example, a halogen, a hydroxy!, a carbony! (such as a carboxyl, an ester, a formy!, or a ketone), a thiocarbonyl (such as a thioester, a thioacetate, or a thioformate), an aikoxyi, a phosphoryi, a phosphonate, a phosphinate, an amino, an amido, an amidine, an imine, a cyano, a nitro, an azido, a sulfhydryl, an alkylthio, a sulfate, a sulfonate, a sulfamoyi, a sulfonamide, a sulfonyl, a heferocyclyl, an aralkyi, or an aromatic or heteroaromatic moiety. It will be understood by those skilled in the art that the moieties substituted on the hydrocarbon chain can themselves be substituted, if appropriate. For instance, the substituents of a substituted alkyl may include substituted and unsubstituted forms of amino, azido, imino, amido, phosphoryi (including phosphonate and phosphinate), sulfonyl (including sulfate, sulfonamide, sulfamoyi and sulfonate), and siiyi groups, as well as ethers, alkylthios, carbonyls (including ketones, aldehydes, carboxyiates, and esters), -CPs, -CN and the like. Exemplary substituted alkyls are described below. Cycioaikyls can be further substituted with alkyls, alkenyls, alkoxys, a!kylthios, aminoaikyls, carbonyi-substituted alky!s, -CF3, -CN, and the like.

The term "aralkyi", as used herein, refers to an alkyl group substituted with an aryl group (e.g., an aromatic or heteroaromatic group).

The terms "aikeny!" and "alkynyl" refer to unsaturated aliphatic groups analogous in length and possible substitution to the alkyls described above, but that contain at least one double or triple bond respectively.

Unless the number of carbons is otherwise specified, "lower alkyl" as used herein means an alkyl group, as defined above, but having from one to ten carbons, for example, from one to six carbon atoms in its backbone structure. Likewise, "lower alkenyi" and "lower alkynyl" have similar chain lengths. In some embodiments, alkyl groups are lower alkyls. In some embodiments, a substituent designated herein as alkyl is a lower alkyl.

The term "aryl" as used herein includes 5-, 6- and 7-membered single-ring aromatic groups that may include from zero to four heteroatoms, for example, benzene, pyrrole, furan, thiophene, imidazole, oxazoie, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazine and pyrimidine, and the like. Those aryl groups having heteroatoms in the ring structure may also be referred to as "aryl heterocycles" or "beteroaromatics". The aromatic ring can be substituted at one or more ring positions with such substituents as described above, for example, halogen, azide, alkyl, aralkyi, alkenyi, alkynyl, cycloalkyl, hydroxyl, amino, nitro, sulfhydryl, imino, amido, phosphionate, phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl, sulfonamide, ketone, aldehyde, ester, heterocyclyl, aromatic or heteroaromatic moieties, -CF3, -CN, or the like. The term "aryl" also includes poiycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjoining rings (the rings are "fused rings") wherein at least one of the rings is aromatic, e.g., the other cyclic rings can be cycioa!kyis, cycloalkenyls, cycioaikynyis, ary!s and/or heterocyciy!s.

The terms "heterocyciy!" or "heterocyclic group" refer to 3- to 10-membered ring structures, for example, 3- to 7-membered rings, whose ring structures include one to four heteroatoms. Heterocycles can also be polycycles. Heterocyclyl groups include, for example, thiophene, thianthrene, furan, pyran, isobenzofuran, chromene, xanthene, phenoxathiin, pyrrole, imidazole, pyrazole, isothiazole, isoxazole, pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole, indole, indazole, purine, quinolizine, isoquino!ine, quinoline, phthalazine, naphthyridine, quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline, phenanthridine, acridine, pyrimidine, phenanthroline, phenazine, phenarsazine, phenothiazine, furazan, phenoxazine, pyrrolidine, oxolane, thiolane, oxazole, piperidine, piperazine, morpholine, lactones, lactams such as azetidinones and pyrrolidinones, suitams, sulfones, and the like. The heterocyclic ring can be substituted at one or more positions with such substituents as described above, as for example, halogen, alkyl, ara!kyl, alkeny!, alkynyl, cycloalkyl, hydroxyl, amino, nitro, sulfhydryl, imino, amido, phosphonate, phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio, su!fonyl, ketone, aldehyde, ester, a heterocyclyl, an aromatic or heteroaromatic moiety, -CF3, -CM, or the like.

The terms "polycyclyl" or "polycyclic group" refer to two or more rings (e.g., cycloalkyls, cycloalkenyls, cycloalkynyls, aryls and/or heterocyclyls) in which two or more carbons are common to two adjoining rings, e.g., the rings are "fused rings". Rings that are joined through non-adjacent atoms are termed "bridged" rings. Each of the rings of the polycycle can be substituted with such substituents as described above, as for example, halogen, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxy!, amino, nitro, sulfhydryl, imino, amido, phosphonate, phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyi, ketone, aldehyde, ester, a heterocyclyl, an aromatic or heteroaromatic moiety, -CF3, -CM, or the like.

The term "carbocycle", as used herein, refers to an aromatic or non-aromatic ring in which each atom of the ring is carbon.

The term "heteroatom" as used herein means an atom of any element other than carbon or hydrogen. Examplery heteroatoms are nitrogen, oxygen, sulfur and phosphorous.

As used herein, the term "nitro" means -NO₂; the term "halogen" designates -F, -CI, -Br or - I; the term "sulfhydryl" means -SH; the term "hydroxyl" means -OH; and the term "sulfonyi" means -SO2-.

It will be understood that "substitution" or "substituted with" includes the implicit proviso that such substitution is in accordance with permitted valence of the substituted atom and the substituent, and that the substitution results in a stable compound, e.g., which does not spontaneously undergo transformation such as by rearrangement, cyciization, elimination, etc.

As used herein, the term "substituted" is contemplated to include all permissible substituents of organic compounds. In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and nonaromatic substituents of organic compounds. Illustrative substituents include, for example, those described hereinabove. The permissible substituents can be one or more and the same or different for appropriate organic compounds. For purposes of this invention, the heteroatoms such as nitrogen may have hydrogen substituents and/or any permissible substituents of organic compounds described herein which satisfy the valencies of the heteroatoms. This invention is not intended to be limited in any manner by the permissible substituents of organic compounds.

A "small hydrophobic" substituent is for example a lower alkyl or a halogen.

By the terms "amino acid residue" and "peptide residue" is meant an amino acid or peptide molecule without the -OH of its carboxyl group. In general, the abbreviations used herein for designating the amino acids and the protective groups are based on recommendations of the lUPAC-IUB Commission on Biochemical Nomenclature (see Biochemistry (1972) 11:1726-1732). For instance, Met, lie, Leu, Ala and Gly represent "residues" of methionine, isoleucine, leucine, alanine and glycine, respectively. By the residue is meant a radical derived from the corresponding alpha-amino acid by eliminating the OH portion of the carboxyl group and the H portion of the alpha-amino group. The term "amino acid side chain" is that part of an amino acid exclusive of the -CH(NH₂)COOH portion, as defined by K. D. Koppie, "Peptides and Amino Acids", W. A. Benjamin Inc., New York and Amsterdam, 1966, pages 2 and 33.

For the most part, the amino acids used in the application of this invention are those naturally occurring amino acids found in proteins, or the naturally occurring anabolic or catabolic products of such amino acids which contain amino and carboxyl groups. Particularly suitable amino acid side chains include side chains selected from those of the following amino acids: glycine, alanine, valine, cysteine, leucine, isoleucine, serine, threonine, methionine, glutamic acid, aspartic acid, glutamine, asparagine, lysine, arginine, proline, histidine, phenylalanine, tyrosine, and tryptophan, and those amino acids and amino acid analogs which have been identified as constituents of peptidylglycan bacterial ceil walls.

The term amino acid residue further includes analogs, derivatives and congeners of any specific amino acid referred to herein, as for instance, the subject compound can include an amino acid analog such as, for example, cyanoalanine, canavanine, djenkolic acid, norieucine, 3-phosphoserine, hornoserine, dihydroxy-phenylalanine, 5-hydroxytryptophan, 1-methylhistidine, 3-methylhistidine, diaminiopirnelic acid, ornithine, or diaminobutyric acid. Other naturally occurring amino acid metabolites or precursors having side chains which are suitable herein will be recognized by those skilled in the art and are included in the scope of the present invention.

Also included are the (D) and (L) stereoisomers of such amino acids when the structure of the amino acid admits of stereoisomeric forms. The configuration of the amino acids and amino acid residues herein are designated by the appropriate symbols (D), (L) or (DL), furthermore when the configuration is not designated the amino acid or residue can have the configuration (D), (L) or (DL). It will be noted that the structure of some of the compounds of this invention includes asymmetric carbon atoms. It is to be understood accordingly that the isomers arising from such asymmetry are included within the scope of this invention. Such isomers can be obtained in substantially pure form by classical separation techniques and by sterically controlled synthesis. For the purposes of this application, unless expressly noted to the contrary, a named amino acid shall be construed to include both the (D) or (L) stereoisomers.

As noted above, certain compounds of the present invention may exist in particular geometric or stereoisomeric forms. The present invention contemplates ail such compounds, including ess- and trans-isomers, R- and S-enantiomers, diastereomers, (D)- isomers, (L)-isomers, the racemic mixtures thereof, and other mixtures thereof, as, failing within the scope of the invention. Additional asymmetric carbon atoms may be present in a substituent such as an alkyl group. Ail such isomers, as well as mixtures thereof, are intended to be included in this invention.

If, for instance, a particular enantiomer of a compound of the present invention is desired, it may be prepared by asymmetric synthesis, or by derivation with a chirai auxiliary, where the resulting diastereomeric mixture is separated and the auxiliary group cleaved to provide the pure desired enantiomers. Alternatively, where the molecule contains a basic functional group, such as amino, or an acidic functional group, such as carboxyl, diastereomeric salts are formed with an appropriate optically-active acid or base, followed by resolution of the diastereomers thus formed by fractional crystallization or chromatographic means well known in the art, and subsequent recovery of the pure enantiomers.

The term "prodrug" as used herein encompasses compounds that, under physiological conditions, are converted into therapeutically active agents. A common method for making a prodrug is to include selected moieties that are hydroiyzed under physiological conditions to reveal the desired molecule. In other embodiments, the prodrug is converted by an enzymatic activity of the host animal.

The term "body mass index" (BMI) is used to mean a statistical measurement which compares a person's weight and height. Though it does not actually measure the percentage of body fat, it is a useful tool to estimate a healthy body weight based on how tall a person is. Body mass index is defined as the individual's body weight divided by the square of their height. The formulas universally used in medicine produce a unit of measure of kg/m².

The term "obesity" is used to mean a condition in which excess body fat has accumulated to such an extent that health may be negatively affected, it is commonly defined as a BMI of about 30 kg/m² or higher. This distinguishes it from being "overweight," as defined by a BMI of between about 25-29.9 kg/m².

For purposes of this invention, the chemical elements are identified in accordance with the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 67th Ed., 1986-87, inside cover. Also for purposes of this invention, the term "hydrocarbon" is contemplated to include ail permissible compounds having at least one hydrogen and one carbon atom, in a broad aspect, the permissible hydrocarbons include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and nonaromatic organic compounds which can be substituted or unsubstituted.

Exemplary Structures

The present invention relates generally to pharmaceutical preparations comprising a multimediator inhibitor agent that both inhibits DPP4 enzymatic activity with a Ki of 10 nM or less and inhibits FAP enzymatic activity with a Ki of 10 nM or less, which multimediator is orally active and is formulated for oral administration at a dosing frequency of 24 hours or greater.

wherein

R¹ represents a naturally or non-naturally occurring amino acid side chain;

R² represents a moiety including two or more overlapping p-orbitals with delocalized electrons, which forms a conjugated system with the nitrogen to which R² is attached such that the nitrogen is a tertiary amine which maintains an electron lone pair under conditions in which the multimediator interacts with DPP4;

R⁵ represents a hydrogen, a lower alkyi, or a -lower a!ky!-haiogen (such as to form a halo-alky! ketone, i.e., -CH3-CI);

R⁷ represents, for each occurrence, hydrogen, or a substituted or unsubstituted lower a!kyl, lower alkenyl, aryl, ara!kyl, cyc!oa!kyl, cyc!oa!kenyi, or heterocycle;

R⁵⁰ represents O or S;

R⁵¹ represents N₃, SH₂, N H₂, N0₂ or OR⁷;

R⁵² represents hydrogen, a lower alkyl, amine, OR⁷, or a pharmaceutically acceptable salt, or R⁵ⁱ and R⁵² taken together with the phosphorous atom to which they are attached complete a heterocyclic ring having from 5 to 8 atoms in the ring structure;

W represents a functional group which reacts with an active site residue of DPP4 and FAP, as for example,—-CN,— -CH=N R⁵,

X represents O or S;

X^I represents a halogen Yi and Y₂ can independently or together be OH, or a group capable of being hydrolyzed to a hydroxy! group, including cyclic derivatives where Yi and Y₂ are connected via a ring having from 5 to 8 atoms in the ring structure (such as pinacol or the like).

In certain embodiments, the ring Z is represented by

wherein

R³ is absent or represents from 1 to 3 ring substituents, each of which can independently be a halogen, a lower alkyl, a lower alkenyl, a lower alkynyl, a carbonyl (such as a carboxyl, an ester, a formate, or a ketone), a thiocarbonyl (such as a thioester, a thioacetate, or a thioformate), an amino, an acyiamino, an amido, a cyano, a nitro, an azido, a sulfate, a sulfonate, a sulfonamide), — (Chbjm— R⁷,— fCH₂)_m-0-R⁷,-(CH₂)_m-S-R⁷;

n represents 1, 2, 3 or 4; and

m represents zero or an integer from 1 to 8,

For example, the ring Z can be

In certain embodiments, R² is an aromatic moiety which forms the conjugated system with the nitrogen to which R² is attached.

In certain embodiments, R² is an aromatic acyl group, to provide a muitimediator is represented in the general formula:

wherein

R³ is absent or represents from 1 to 3 ring substituents, each of which can independently be a halogen, a lower alkyl, a lower alkenyi, a lower aikynyi, a carbonyl (such as a carboxyi, an ester, a formate, or a ketone), a thiocarbonyl (such as a thioester, a thioacetate, or a thioformate), an amino, an acylamino, an amido, a cyano, a nitro, an azido, a sulfate, a sulfonate, a sulfonamido, — (ChkJm— R⁷,— iCH₂)_m— O— R⁷,— (C^— S—R⁷;

R⁴ represents an aromatic group, which along with the acyl group, forms the conjugated system with the nitrogen;

n represents 1, 2, 3 or 4; and

m represents zero or an integer from 1 to 8.

In certain embodiments, the aromatic moiety is a hydroxylated benzene moiety, substituted with from 1 to 5 hydroxyl groups (for example, 2 hydroxy! groups). In certain embodiments, the aromatic moiety is a haiogenated benzene moiety, substituted with from 1 to 5 halogen groups, for example with 2 halogen groups, which halogen groups are for example CI, Br or F.

In certain embodiments of the muitimediator, R¹ is a hydrogen or a small hydrophobic group.

In certain embodiments, the muitimediator is represented in the general formula:

wherein

R¹ is hydrogen or a lower akyi;

R^:3a and R^3b are each hydrogen, or R^3a is hydrogen and R^:3 is -OH, or R^:3a and R^3b each independently are a halogen, for example, F; and

R⁶ represents 1, 2 or 3 hydroxy! groups, or R^b represents 1, 2 or 3 halogens, for example, F.

Pharmaceutical Formulations

The inhibitors can be administered in various forms, depending on the disorder to be treated and the age, condition and body weight of the patient, as is well known in the art. For example, where the compounds are to be administered orally, they may be formulated as tablets, capsules, granules, powders or syrups; or for parenteral administration, they may be formulated as injections (intravenous, intramuscular or subcutaneous), drop infusion preparations or suppositories. For application by the ophthalmic mucous membrane route, they may be formulated as eyedrops or eye ointments. These formulations can be prepared by conventional means, and, if desired, the active ingredient may be mixed with any conventional additive, such as an excipient, a binder, a disintegrating agent, a lubricant, a corrigent, a solubilizing agent, a suspension aid, an emulsifying agent or a coating agent. Although the dosage will vary depending on the symptoms, age and body weight of the patient, the nature and severity of the disorder to be treated or prevented, the route of administration and the form of the drug, in general, a daily dosage of from 0.01 to 2000 mg of the compound is recommended for an adult human patient, and this may be administered in a single dose or in divided doses.

Glucose metabolism can be altered, and symptoms associated with type 11 diabetes can be decreased or eliminated, in accordance with a "timed" administration of muitimediator inhibitors wherein one or more appropriate indices for glucose metabolism and/or type II diabetes can be used to assess effectiveness of the treatment (dosage and/or timing): e.g. glucose tolerance, glucose level, insulin level, insulin sensitivity, glycosylated hemoglobin.

An effective time for administering muitimediator inhibitors can be accomplished by routine experiment as described below, using one or more groups of animals (for example, at least 5 animals per group).

The precise time of administration and/or amount of muitimediator inhibitor that will yield the most effective results in terms of efficacy of treatment in a given patient will depend upon the activity, pharmacokinetics, and bioavailability of a particular compound, physiological condition of the patient (including age, sex, disease type and stage, general physical condition, responsiveness to a given dosage and type of medication), route of administration, etc. However, the above guidelines can be used as the basis for fine-tuning the treatment, e.g., determining the optimum time and/or amount of administration, which will require no more than routine experimentation consisting of monitoring the subject and adjusting the dosage and/or timing.

While the subject is being treated, glucose metabolism is monitored by measuring one or more of the relevant indices at predetermined times during a 24-hour period. Treatment (amounts, times of administration and type of medication) may be adjusted (optimized) according to the results of such monitoring. The patient is periodically reevaluated to determine extent of improvement by measuring the same parameters, the first such reeva!uation typically occurring at the end of four weeks from the onset of therapy, and subsequent reevaluations occurring every 4 to 8 weeks during therapy and then every 3 months thereafter. Therapy may continue for several months or even years with six months being a typical length of therapy for humans.

Adjustments to the amounf(s) of drug(s) administered and possibly to the time of administration may be made based on these reevaluations. For example, if after 4 weeks of treatment one of the metabolic indices has not improved but at least one other one has, the dose could be increased by 1/3 without changing the time of administration.

Treatment can be initiated with smaller dosages which are less than the optimum dose of the compound. Thereafter, the dosage should be increased by small increments until the optimum effect under the circumstances is reached. For convenience, the total daily dosage may be divided and administered in portions during the day if desired.

The phrase "therapeutically-effective amount" as used herein means that amount of, e.g., a multimediator inhibitor(s), which is effective for producing some desired therapeutic effect by inhibiting, for example, the proteolysis of a peptide hormone at a reasonable benefit/risk ratio applicable to any medical treatment.

The phrase "pharmaceutically acceptable" is employed herein to refer to those multimediator inhibitors, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.

The phrase "pharmaceutically-acceptable carrier" as used herein means a pharmaceutically-acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material, involved in carrying or transporting the subject chemical from one organ, or portion of the body, to another organ, or portion of the body. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient. Some examples of materials which can serve as pharmaceuticaliy-acceptable carriers include: (1) sugars, such as lactose, glucose and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose, and its derivatives, such as sodium carboxymethyi cellulose, ethyl cellulose and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitoi and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffering agents, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethyl alcohol; (20) phosphate buffer solutions; and (21) other non-toxic compatible substances employed in pharmaceutical formulations.

The term "pharmaceuticaliy-acceptable salts" refers to the relatively non-toxic, inorganic and organic acid addition salts of multimediator inhibitors. These salts can be prepared in situ during the final isolation and purification of the multimediator inhibitors, or by separately reacting a purified multimediator inhibitor in its free base form with a suitable organic or inorganic acid, and isolating the salt thus formed. Representative salts include the hydrobromide, hydrochloride, sulfate, bisuifate, phosphate, nitrate, acetate, valerate, oleate, palmitate, stearate, laurate, benzoate, lactate, phosphate, tosylate, citrate, maieate, fumarate, succinate, tartrate, napthyiate, mesylate, glucoheptonate, iacfobionafe, and laurylsulphonate salts and the like. (See, for example, Berge et al. (1977) "Pharmaceutical Salts", J. Pharm. Sci. 66:1-19)

in other cases, the multimediator inhibitor useful in the methods of the present invention may contain one or more acidic functional groups and, thus, are capable of forming pharmaceuticaliy-acceptable salts with pharmaceuticaliy-acceptable bases. The term "pharmaceuticaliy-acceptable salts" in these instances refers to the relatively non-toxic, inorganic and organic base addition salts of a multimediator inhibitor(s). These salts can likewise be prepared in situ during the final isolation and purification of the multimediator inhibitor(s), or by separately reacting the purified multimediator inhibitor(s) in its free acid form with a suitable base, such as the hydroxide, carbonate or bicarbonate of a pharmaceuticaliy-acceptable metal cation, with ammonia, or with a pharmaceuticaliy- acceptable organic primary, secondary or tertiary amine Representative alkali or alkaline earth salts include the lithium, sodium, potassium, calcium, magnesium, and aluminum salts and the like. Representative organic amines useful for the formation of base addition salts include ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine and the like (see, for example, Berge et al., supra). Wetting agents, emuisifiers and lubricants, such as sodium iauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the compositions.

Examples of pharmaceutically-acceptable antioxidants include: (1) water soluble antioxidants, such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite and the like; (2) oil-soluble antioxidants, such as ascorbyi palmitate, butylated hydroxyanisole (BHA), butyiated hydroxytoluene (BHT), lecithin, propyl gailate, alpha-tocopherol, and the like; and (3) metal chelating agents, such as citric acid, ethyienediamine tetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, and the like.

Formulations useful in the methods of the present invention include those suitable for oral, nasal, topical (including buccal and sublingual), rectal, vaginal, aerosol and/or parenteral administration. The formulations may conveniently be presented in unit dosage form and may be prepared by any methods well known in the art of pharmacy. The amount of active ingredient which can be combined with a carrier material to produce a single dosage form will vary depending upon the host being treated, the particular mode of administration. The amount of active ingredient which can be combined with a carrier material to produce a single dosage form will generally be that amount of the compound which produces a therapeutic effect. Generally, out of one hundred percent, this amount will range from about 1 percent to about ninety-nine percent of active ingredient, for example, from about 5 percent to about 70 percent, or from about 10 percent to about 30 percent.

Methods of preparing these formulations or compositions include the step of bringing into association a multimediator inhibitor(s) with the carrier and, optionally, one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association a multimediator inhibitor with liquid carriers, or finely divided solid carriers, or both, and then, if necessary, shaping the product.

Formulations suitable for oral administration may be in the form of capsules, cachets, pills, tablets, lozenges (using a flavored basis, usually sucrose and acacia or tragacanth), powders, granules, or as a solution or a suspension in an aqueous or non-aqueous liquid, or as an oil-in-water or water-in-oil liquid emulsion, or as an elixir or syrup, or as pastilles (using an inert base, such as gelatin and glycerin, or sucrose and acacia) and/or as mouth washes and the like, each containing a predetermined amount of a multimediator inhibitor(s) as an active ingredient. A compound may also be administered as a bolus, electuary or paste. Irs solid dosage forms for oral administration (capsules, tablets, pills, dragees, powders, granules and the like), the active ingredient is mixed with one or more pharmaceutically- acceptable carriers, such as sodium citrate or dicalcium phosphate, and/or any of the following: (1) fillers or extenders, such as starches, lactose, sucrose, glucose, mannitol, and/or silicic acid; (2) binders, such as, for example, carboxymethyiceliulose, alginates, gelatin, polyvinyl pyrroiidone, sucrose and/or acacia; (3) humectants, such as glycerol; (4) disintegrating agents, such as agar-agar, calcium carbonate, potato or tapioca starch, aiginic acid, certain silicates, and sodium carbonate; (5) solution retarding agents, such as paraffin; (6) absorption accelerators, such as quaternary ammonium compounds; (7) wetting agents, such as, for example, acetyl alcohol and glycerol monostearate; (8) absorbents, such as kaolin and bentonite clay; (9) lubricants, such a talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof; and (10) coloring agents. In the case of capsules, tablets and pills, the pharmaceutical compositions may also comprise buffering agents. Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugars, as well as high molecular weight polyethylene glycols and the l ike.

A tablet may be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared using binder (for example, gelatin or hydroxypropylmethyl cellulose), lubricant, inert diluent, preservative, disintegrant (for example, sodium starch glycoiate or cross-linked sodium carboxymefhyi cellulose), surface- active or dispersing agent. Molded tablets may be made by molding in a suitable machine a mixture of the powdered peptide or peptidomimetic moistened with an inert liquid diluent.

Tablets, and other solid dosage forms, such as dragees, capsules, pills and granules, may optionally be scored or prepared with coatings and shells, such as enteric coatings and other coatings well known in the pharmaceutical-formulating art. They may also be formulated so as to provide slow or controlled release of the active ingredient therein using, for example, hydroxypropylmethyl cellulose in varying proportions to provide the desired release profile, other polymer matrices, liposomes and/or microspheres. They may be sterilized by, for example, filtration through a bacteria-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved in sterile water, or some other sterile injectable medium immediately before use. These compositions may also optionally contain opacifying agents and may be of a composition that they release the active ingredient(s) only, or in a certain portion of the gastrointestinal tract, optionally, in a delayed manner. Examples of embedding compositions which can be used include polymeric substances and waxes. The active ingredient can also be in micro-encapsulated form, if appropriate, with one or more of the above-described excipients. Liquid dosage forms for orai administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs, in addition to the active ingredient, the liquid dosage forms may contain inert diluents commonly used in the art, such as, for example, water or other solvents, solubilizing agents and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor and sesame oils), glycerol, tetrahydrofuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof.

Besides inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, coloring, perfuming and preservative agents.

Suspensions, in addition to the active multimediator inhibitor(s) may contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanfh, and mixtures thereof.

Formulations for rectal or vaginal administration may be presented as a suppository, which may be prepared by mixing one or more multimediator inhibitor(s) with one or more suitable nonirrifating excipients or carriers comprising, for example, cocoa butter, polyethylene glycol, a suppository wax or a salicylate, and which is solid at room temperature, but liquid at body temperature and, therefore, will melt in the rectum or vaginal cavity and release the active agent.

Formulations which are suitable for vaginal administration also include pessaries, tampons, creams, gels, pastes, foams or spray formulations containing such carriers as are known in the art to be appropriate.

Dosage forms for the topical or transdermal administration of a multimediator inhibitor(s) include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and inhalants. The active component may be mixed under sterile conditions with a pharmaceutically-acceptable carrier, and with any preservatives, buffers, or propeilants which may be required.

The ointments, pastes, creams and gels may contain, in addition to multimediator inhibitor(s), excipients, such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.

Powders and sprays can contain, in addition to a multimediator inhibitor(s), excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and poiyamide powder, or mixtures of these substances. Sprays can additionally contain customary propeliants, such as chiorofiuorohydrocarbons and volatile unsubstituted hydrocarbons, such as butane and propane.

The multimediator inhibitor(s) can be alternatively administered by aerosol. This is accomplished by preparing an aqueous aerosol, liposomal preparation or solid particles containing the compound. A nonaqueous (e.g.. fiuorocarbon propellant) suspension could be used. Sonic nebulizers minimize exposing the agent to shear, which can result in degradation of the compound.

Ordinarily, an aqueous aerosol is made by formulating an aqueous solution or suspension of the agent together with conventional pharmaceutically acceptable carriers and stabilizers. The carriers and stabilizers vary with the requirements of the particular compound, but typically include nonionic surfactants (Tweens, Piuronics, or polyethylene glycol), innocuous proteins like serum albumin, sorbitan esters, oleic acid, lecithin, amino acids such as glycine, buffers, salts, sugars or sugar alcohols. Aerosols generally are prepared from isotonic solutions.

Transdermal patches have the added advantage of providing controlled delivery of a multimediator inhibitor(s) to the body. Such dosage forms can be made by dissolving or dispersing the agent in the proper medium. Absorption enhancers can also be used to increase the flux of the peptldomimetic across the skin. The rate of such flux can be controlled by either providing a rate controlling membrane or dispersing the peptldomimetic in a polymer matrix or gel.

Ophthalmic formulations, eye ointments, powders, solutions and the like, are also contemplated as being within the scope of this invention.

Pharmaceutical compositions of this invention suitable for parenteral administration comprise one or more multimediator inhibitor(s) in combination with one or more pharmaceutically-acceptable sterile isotonic aqueous or nonaqueous solutions, dispersions, suspensions or emulsions, or sterile powders which may be reconstituted into sterile injectable solutions or dispersions just prior to use, which may contain antioxidants, buffers, bacteriostats, solutes which render the formulation isotonic with the blood of the intended recipient or suspending or thickening agents.

Examples of suitable aqueous and nonaqueous carriers which may be employed in the pharmaceutical compositions of the invention include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, and the like), and suitable mixtures thereof, vegetable oils, such as olive oil, and injectable organic esters, such as ethyl oleate. Proper fluidity can be maintained, for example, by the use of coating materials, such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants.

These compositions may also contain adjuvants such as preservatives, wetting agents, emulsifying agents and dispersing agents. Prevention of the action of microorganisms may be ensured by the inclusion of various antibacterial and antifungal agents, for example, paraben, chlorobutanol, phenol sorbic acid, and the like. It may also be desirable to include isotonic agents, such as sugars, sodium chloride, and the like into the compositions, in addition, prolonged absorption of the injectable pharmaceutical form may be brought about by the inclusion of agents which delay absorption such as aluminum monostearate and gelatin.

In some cases, in order to prolong the effect of a drug, it is desirable to slow the absorption of the drug from subcutaneous or intramuscular injection. This may be accomplished by the use of a liquid suspension of crystalline or amorphous material having poor water solubility. The rate of absorption of the drug then depends upon its rate of dissolution which, in turn, may depend upon crystal size and crystalline form. Alternatively, delayed absorption of a parenteraliy-administered drug form is accomplished by dissolving or suspending the drug in an oil vehicle.

Injectable depot forms are made by forming microencapsu!e matrices of multimediator inhibitor(s) in biodegradable polymers such as polylactide-polyglycolide. Depending on the ratio of drug to polymer, and the nature of the particular polymer employed, the rate of drug release can be controlled. Examples of other biodegradable polymers include po!yforthoesters) and poly(anhydrides). Depot injectable formulations are also prepared by entrapping the drug in liposomes or microemulsions which are compatible with body tissue.

When the multimediator inhibitor(s) of the present invention are administered as pharmaceuticals, to humans and animals, they can be given per se or as a pharmaceutical composition containing, for example, 0.1 to 99.5% or 0.5 to 90% of active ingredient in combination with a pharmaceutically acceptable carrier.

The preparations of agents may be given orally, parenterally, topically, or rectally. They are of course given by forms suitable for each administration route. For example, they are administered in tablets or capsule form, by injection, inhalation, eye lotion, ointment, suppository, etc. administration by injection, infusion or inhalation; topical by lotion or ointment; and rectal by suppositories.

The phrases "parenteral administration" and "administered parenterally" as used herein means modes of administration other than enteral and topical administration, usually by injection, and includes, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticulare, subcapsular, subarachnoid, intraspinal and intrasternal injection and infusion.

The phrases "systemic administration," "administered systemicaliy," "peripheral administration" and "administered peripherally" as used herein mean the administration of a multimediator inhibitor, drug or other material other than directly into the central nervous system, such that it enters the patient's system and, thus, is subject to metabolism and other like processes, for example, subcutaneous administration.

These multimediator inhibitor{s) may be administered to humans and other animals for therapy by any suitable route of administration, including orally, nasally, as by, for example, a spray, rectally, intravaginaily, parenteraliy, intracisternaliy and topically, as by powders, ointments or drops, including buccaliy and sublingually.

Regardless of the route of administration selected, the multimediator inhibitor(s), which may be used in a suitable hydrated form, and/or the pharmaceutical compositions of the present invention, are formulated into pharmaceutica!ly-acceptab!e dosage forms by conventional methods known to those of skill in the art.

Actual dosage levels of the active ingredients in the pharmaceutical compositions of this invention may be varied so as to obtain an amount of the active ingredient which is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient.

In certain embodiments, the invention relates to any one of the aforementioned methods, wherein the compound is administered to the mammal orally.

In certain embodiments, the invention relates to any one of the aforementioned methods, wherein the compound is administered to the mammal chronically. In certain embodiments, chronic administration or chronic dosing takes place over a period of time. In certain embodiments, the period of time is greater than about 2 weeks, greater than about 3 weeks, greater than about 4 weeks, greater than about 5 weeks, greater than about 6 weeks, greater than about 7 weeks, greater than about 8 weeks, greater than about 9 weeks, or greater than about 10 weeks. In certain embodiments, a chronic dose is about 0.1 mg/kg/day, about 0.2 mg/kg/day, about 0.3 mg/kg/day, about 0.4 mg/kg/day, about 0.5 mg/kg/day, about 0.6 mg/kg/day, about 0.7 mg/kg/day, about 0.8 mg/kg/day, about 0.9 mg/kg/day, about 1 mg/kg/day, about 1.5 mg/kg/day, about 2 mg/kg/day, about 2.5 mg/kg/day, about 3 mg/kg/day, about 3.5 mg/kg/day, about 4 mg/kg/day, about 4.5 mg/kg/day, or about 5 mg/kg/day over a period of time. In certain embodiments, a chronic dose is about 0.5

about 1 moie/kg/day, about 1.5 about 2

about 3 μϊ-nole/kg/day, about 3.5

about 4

about 5.5 μΓηοΙθ/Ί<§/¾ν, about 6 ΓηοΙθ/!< /ά3ν, about 6.5

about 7.5

about 8.5 μηηοΙθ/Ι , τΐ3ν, about 9

about 9.5 ϊτιοΐ6/1 §/οΐ3ν, about 10 πιοΐ6/Ι §/οΐ3ν, about 11 μϊ-nole/kg/day, about 12 πιοΐ6/Ι<§/ά3ν, about 13

about 14 μπιοίθ/!¾/εί3ν, or about 15

over a period of time.

Exemplary Therapeutic Uses

The multimediator inhibitor of the present invention can be used for a variety of different disorders, particularly those in which elongation of the serum haif-life (lives) or one or more substrates for DPP4 and FAP provides therapeutic benefit. For instance, the pharmaceutical preparations of the present invention can be used for the following:

* A method for reducing a blood glucose level of a subject having an elevated blood glucose level or preventing or delaying the onset of an elevated blood glucose level in a subject at risk for developing an elevated glucose level;

* A method for improving insulin sensitivity in a subject with insulin resistance or delaying the onset of insulin resistance in a subject at risk for developing insulin resistance;

* A method for improving glucose tolerance in a subject with impaired glucose tolerance;

* A method of treating at least one metabolic disorder in a subject or preventing or delaying the onset of at least one metabolic disorder, such as selected from prediabetes, diabetes, metabolic syndrome, obesity, diabetic dyslipidemia, hyperlipdemia, hypertension, hypertriglyceridemia, hyperfattyacidemia, hypercholesterolemia, and hyperinsulinemia;

* A method for treating fatty liver disease in a subject, such as non-alcoholic fatty liver disease (NAFLD), alcoholic fatty liver disease, alcoholic steatohepatitis, or nonalcoholic steatohepatitis (NASH);

* A method of reducing liver triglycerides;

* A method of treating a patient having a cardiovascular (CV) and/or renal microvascular disease, particularly where the multimediator inhibitor is both cardioprotective and renoprofective. Such treatments can be used to protect against, delay the occurrence of, delay the progression of and/or reduce the risk of such diseases as CV morbidity, premature CV mortality, renal morbidity and premature renal mortality. For instance, the subject pharmaceutical preparations can be used as part of treatments intended to protect against, reduce the risk of and/or delays the occurrence of: a cardio- or cerebrovascular disease or event selected from the group consisting of cardiovascular (CV) death selected from the group consisting of fatal stroke, fatal myocardial infarction, fatal heart failure, cardiogenic shock and sudden death, non-fatal stroke and non-fatal myocardial infarction (Ml) (with or without silent M l), and, optionally, hospitalization, wherein said hospitalization is for unstable angina pectoris, stable angina pectoris, transient ischemic attack, coronary revascularization procedures, peripheral revascularization or congestive heart failure, and/or (b) protects against, reduces the risk of, delays the progression of and/or delays the occurrence of a renal microvascular disease selected from the group consisting of: abuminuria, chronic kidney disease (CKD), renal impairment, renal death, end-stage renal disease and loss in estimated glomerular filtration rate;

A method of slowing the progression of, delaying the onset of or treating a metabolic disorder or disease, such as type 1 diabetes mellitus, type 2 diabetes mellitus, impaired glucose tolerance (IGT), impaired fasting blood glucose (I FG), hyperglycemia, postprandial hyperglycemia, postabsorptive hyperglycemia, latent autoimmune diabetes in adults (LADA), overweight, obesity, dyslipidemia, hyperiipidemia, hypercholesterolemia, hypertriglyceridemia, hyperis!EFA-emia, postprandial lipemia, hypertension, atherosclerosis, endothelial dysfunction, osteoporosis, chronic systemic inflammation, non alcoholic fatty liver disease (NAFLD), retinopathy, neuropathy, nephropathy, nephrotic syndrome, polycystic ovarian syndrome, and/or metabolic syndrome; improving and/or maintaining glycemic control and/or for reducing of fasting plasma glucose, of postprandial plasma glucose, of postabsorptive plasma glucose and/or of glycosylated hemoglobin HbAlc; slowing, delaying or reversing progression from prediabetes, impaired glucose tolerance (IGT), impaired fasting blood glucose (I FG), insulin resistance and/or from metabolic syndrome to type 2 diabetes mellitus; reducing the risk of, slowing the progression of, delaying the onset of or treating complications of diabetes mellitus selected from the group consisting of micro- and macrovascular diseases, nephropathy, micro- or macroalbuminuria, proteinuria, nephrotic syndrome, retinopathy, cataracts, neuropathy, learning or memory impairment, neurodegenerative or cognitive disorders, cardio- or cerebrovascular diseases, tissue ischaemia, diabetic foot or ulcer, atherosclerosis, hypertension, endothelial dysfunction, myocardial infarction, acute coronary syndrome, unstable angina pectoris, stable angina pectoris, peripheral arterial occlusive disease, cardiomyopathy, heart failure, heart rhythm disorders, vascular restenosis, and stroke; reducing body weight and/or body fat and/or liver fat and/or intra- myocelluiar fat or facilitating a reduction in body weight and/or body fat and/or liver fat and/or intra-myocellular fat; slowing, delaying the onset of or treating the degeneration of pancreatic beta cells and/or the decline of the functionality of pancreatic beta cells and/or for improving, preserving and/or restoring the functionality of pancreatic beta ceils and/or stimulating and/or restoring or protecting the functionality of pancreatic insulin secretion; slowing, delaying the onset of or treating non alcoholic fatty liver disease (IMAFLD) including hepatic steatosis, non-alcoholic steatohepatitis (NASH) and/or liver fibrosis; slowing the progression of, delaying the onset of or treating type 2 diabetes with failure to conventional antidiabetic mono- or combination therapy; achieving a reduction in the dose of conventional antidiabetic medication required for adequate therapeutic effect; reducing the risk for adverse effects associated with conventional antidiabetic medication; and maintaining and/or improving the insulin sensitivity and/or for treating hyperinsulinemia and/or insulin resistance;

* A method for protecting against, delaying the occurrence of, delaying the progression of and/or reducing the risk of accelerated cognitive decline or impairment, dementia, and/or depressive, mood or anxiety disorders;

* A method for inducing satiety in a subject; or

* A method for reducing fat intake, bodyweight, and/or body fat in a subject

In another aspect, the invention includes a method of treating a subject for a cardiovascular or pulmonary disease or disorder, the method comprising administering a therapeutically effective amount of a multimediator inhibitor to the subject. Cardiovascular and pulmonary diseases and disorders that can be treated by the methods of the invention include, but are not limited to, cardiac or pulmonary fibrosis, cardiac hypertrophy, aortic aneurism, pulmonary hypertension, hypertensive heart disease, arterial inflammation, inflammatory heart disease, endocarditis, inflammatory cardiomegaly, myocarditis, valvular heart disease, cerebrovascular disease, angina, peripheral and coronary artery disease, heart failure, cor pulmonale, myocarditis, cardiomyopathy, atherosclerosis, congenital heart disease, rheumatic heart disease, myocardial infarction, ischemia-reperfusion injury, cardiac dysrhythmia, inflammatory vascular injury, atrial fibrillation, ventricular tachycardia, Brugada syndrome, preeclampsia, angioplasty, restenosis, vascular surgery, cardiac surgery, and cardiac transplantation. By "therapeutically effective dose or amount" in this context is intended an amount that, when administered as described herein, brings about a positive therapeutic response, such as improved recovery from a cardiovascular or pulmonary disease or disorder. Improved recovery may include improved cardiac repair, increased cardiac contractility, increased cardiac output, reduced pressure overload -induced cardiac dysfunction, decreased fibrosis or hypertrophy, decreased infarct size, decreased inflammation, or decreased mortality. Additionally, a therapeutically effective dose or amount may result in clinical improvement in a patient having a cardiovascular or pulmonary disease or disorder as evidenced, for example, by increased exercise toierance/capacity, decreased fluid retention, decreased dyspnea, and/or improved results on quantitative tests of cardiac function (e.g., ejection fraction, exercise capacity).

"Cardiovascular and pulmonary diseases and disorders" include, but are not limited to, cardiac or pulmonary fibrosis, cardiac hypertrophy, aortic aneurism, pulmonary hypertension, hypertensive heart disease, arterial inflammation, inflammatory heart disease, endocarditis, inflammatory cardiomegaly, myocarditis, valvular heart disease, cerebrovascular disease, angina, peripheral and coronary artery disease, heart failure, cor pulmonale, myocarditis, cardiomyopathy, atherosclerosis, congenital heart disease, rheumatic heart disease, myocardial infarction, ischemia-reperfusion injury, cardiac dysrhythmia, inflammatory vascular injury, atrial fibrillation, ventricular tachycardia, Brugada syndrome, preeclampsia, angioplasty, restenosis, vascular surgery, cardiac surgery, and cardiac transplantation.

In certain embodiments, the invention relates to any one of the aforementioned methods, wherein the subject is a mammal.

In certain embodiments, the mammal is a primate, equine, canine, feline, or bovine.

In certain embodiments, the invention relates to any one of the aforementioned methods, wherein the mammal is a human.

In certain embodiments, the invention relates to any one of the aforementioned methods, wherein the mammal is an overweight human. In certain embodiments, the invention relates to any one of the aforementioned methods, wherein the mammal is an obese human.

In certain embodiments, the invention relates to any one of the aforementioned methods, wherein the mammal is a human with a B ! from about 25 kg/m2 to about 50 kg/m2. in certain embodiments, the invention relates to any one of the aforementioned methods, wherein the mammal is a human with a BMI from about 35 kg/m2 to about 50 kg/m2. in certain embodiments, the invention relates to any one of the aforementioned methods, wherein the mammal is a human with a BMI from about 30 kg/m2 to about 50 kg/m2. In certain embodiments, the invention relates to any one of the aforementioned methods, wherein the mammal is a human with a BMI of about 26 kg/m2, about 28 about kg/m2, about 30 kg/m2, about 32 kg/m2, about 34 kg/m2, about 36 kg/m2, about 38 kg/m2, about 40 kg/m2, about 42 kg/m2, about 44 kg/m2, about 46 kg/m2, or about 48 kg/m2. Combination Therapy Overview

As mentioned above, one aspect of the present invention relates to combination therapy - that is, a method comprising the step of administering a first compound and a second compound, wherein the first compound inhibits FAP; and the second compound inhibits DPPiV, This type of therapy is advantageous because the co-administration of active ingredients achieves a therapeutic effect that is greater than the therapeutic effect achieved by administration of oniy a single therapeutic agent.

In certain embodiments, the co-administration of two or more therapeutic agents achieves a therapeutic effect that is greater than the therapeutic effect achieved by administration of only a single therapeutic agent. In this regard, the combination therapies are efficacious. The therapeutic effect of one therapeutic agent is augmented by the coadministration of another therapeutic agent.

In certain embodiments, the co-administration of two or more therapeutic agents achieves a therapeutic effect that is equal to about the sum of the therapeutic effects achieved by administration of each single therapeutic agent. In these embodiments, the combination therapies are said to be "additive."

In certain embodiments, the co-administration of two or more therapeutic agents achieves a synergistic effect, i.e., a therapeutic effect that is greater than the sum of the therapeutic effects of the individual components of the combination.

The active ingredients that comprise a combination therapy may be administered together via a single dosage form or by separate administration of each active agent. In certain embodiments, the first and second therapeutic agents are administered in a single dosage form. The agents may be formulated info a single tablet, pill, capsule, or solution for parenteral administration and the like.

In certain embodiments, the therapeutic agents are administered in a single dosage form, wherein each individual therapeutic agent is isolated from the other therapeutic agent(s). Formulating the dosage forms in such a way assists in maintaining the structural integrity of potentially reactive therapeutic agents until they are administered. A formulation of this type may be useful during production and for long-term storage of the dosage form, in certain embodiments, the therapeutic agents may comprise segregated regions or distinct caplets or the like housed within a capsule, in certain embodiments, the therapeutic agents are provided in isolated layers comprised by a tablet.

Alternatively, the therapeutic agents may be administered as separate compositions, e.g., as separate tablets or solutions. One or more active agent may be administered at the same time as the other active agent(s) or the active agents may be administered intermittently. The length of time between administrations of the therapeutic agents may be adjusted to achieve the desired therapeutic effect. In certain instances, one or more therapeutic agent(s) may be administered only a few minutes (e.g., about 1, 2, 5, 10, 30, or 60 min) after administration of the other therapeutic agent(s). Alternatively, one or more therapeutic agentfs) may be administered several hours (e.g., about 2, 4, 6, 10, 12, 24, or 36 h) after administration of the other therapeutic agent(s). In certain embodiments, it may be advantageous to administer more than one dosage of one or more therapeutic agent(s) between administrations of the remaining therapeutic agent(s). For example, one therapeutic agent may be administered at 2 hours and then again at 10 hours following administration of the other therapeutic agent(s). importantly, it is required that the therapeutic effects of each active ingredient overlap for at least a portion of the duration of each therapeutic agent so that the overall therapeutic effect of the combination therapy is attributable in part to the combined or synergistic effects of the combination therapy.

The dosage of the active agents will generally be dependent upon a number of factors including pharmacodynamic characteristics of each agent of the combination, mode and route of administration of active agentfs), the health of the patient being treated, the extent of treatment desired, the nature and kind of concurrent therapy, if any, and the frequency of treatment and the nature of the effect desired, in general, dosage ranges of the active agents often range from about 0.001 to about 250 mg/kg body weight per day. For a normal adult having a body weight of about 70 kg, a dosage may range from about 0.1 to about 25 mg/kg body weight. However, some variability in this general dosage range may be required depending upon the age and weight of the subject being treated, the intended route of administration, the particular agent being administered and the like. Since two or more different active agents are being used together in a combination therapy, the potency of each agent and the interactive effects achieved using them together must be considered. Importantly, the determination of dosage ranges and optimal dosages for a particular mammal is also well within the ability of one of ordinary skill in the art having the benefit of the instant disclosure.

In certain embodiments, it may be advantageous for the pharmaceutical combination to have a relatively large amount of the first component compared to the second component. In certain instances, the ratio of the first active agent to second active agent is about 200:1, 190:1, 180:1, 170:1, 160:1, 150:1, 140:1, 130:1, 120:1, 110:1, 100:1, 90:1, 80:1, 70:1, 60:1, 50:1, 40:1, 30:1, 20:1, 15:1, 10:1, 9:1, 8:1, 7:1, 6:1, or 5:1. in certain embodiments, it may be preferable to have a more equal distribution of pharmaceutical agents. In certain instances, the ratio of the first active agent to the second active agent is about 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, or 1:4. in certain embodiments, it may be advantageous for the pharmaceutical combination to have a relatively large amount of the second component compared to the first component, in certain instances, the ratio of the second active agent to the first active agent is about 30:1, 20:1, 15:1, 10:1, 9:1, 8:1, 7:1, 6:1, or 5:1. In certain instances, the ratio of the second active agent to first active agent is about 100:1, 90:1, 80:1, 70:1, 60:1, 50:1, or 40:1. In certain instances, the ratio of the second active agent to first active agent is about 200:1, 190:1, 180:1, 170:1, 160:1, 150:1, 140:1, 130:1, 120:1, or 110:1. Importantly, a composition comprising any of the above-identified combinations of first therapeutic agent and second therapeutic agent may be administered in divided doses about 1, 2, 3, 4, 5, 6, or more times per day or in a form that will provide a rate of release effective to attain the desired results. In one embodiment, the dosage form contains both the first and second active agents, in one embodiment, the dosage form only has to be administered one time per day and the dosage form contains both the first and second active agents.

For example, a formulation intended for oral administration to humans may contain from about 0.1 mg to about 5 g of the first therapeutic agent and about 0.1 mg to about 5 g of the second therapeutic agent, both of which are compounded with an appropriate and convenient amount of carrier material varying from about 5 to about 95 percent of the total composition. Unit dosages will generally contain between about 0.5 mg to about 1500 mg of the first therapeutic agent and 0.5 mg to about 1500 mg of the second therapeutic agent. in certain embodiments, the dosage is about 25 mg, 50 mg, 100 mg, 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 800 mg, or 1000 mg, etc., up to about 1500 mg of the first therapeutic agent, in certain embodiments, the dosage is about 25 mg, 50 mg, 100 mg, 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 800 mg, or 1000 mg, etc., up to about 1500 mg of the second therapeutic agent. in certain embodiments, the dosage form contains the first and the second active agents, in certain embodiments, the dosage form only has to be administered one time per day and the dosage form contains the first and the second active agents.

Synergism and Augmentation

The term "synergistic" refers to a combination which is more effective than the additive effects of any two or more single agents. A synergistic effect permits the effective treatment of a disease using iower amounts (doses) of individual therapy. The iower doses result in iower toxicity without reduced efficacy, in addition, a synergistic effect can result in improved efficacy. Finally, synergy may result in an improved avoidance or reduction of disease as compared to any single therapy.

Combination therapy can allow for the product of iower doses of the first therapeutic or the second therapeutic agent (referred to as "apparent one-way synergy" herein), or Iower doses of both therapeutic agents (referred to as "two-way synergy" herein) than would normally be required when either drug is used alone.

In certain embodiments, the synergism exhibited between one or more therapeutic agent(s) and the remaining therapeutic agent(s) is such that the dosage of one of the therapeutic agents would be sub-therapeutic if administered without the dosage of the other therapeutic agents.

The terms "augmentation" or "augment" refer to combinations where one of the compounds increases or enhances therapeutic effects of another compound or compounds administered to a patient, in some instances, augmentation can result in improving the efficacy, tolerability, or safety, or any combination thereof, of a particular therapy.

In certain embodiments, the present invention relates to a pharmaceutical composition comprising a therapeutically effective dose of one or more therapeutic agent(s) together with a dose of another therapeutic agent effective to augment the therapeutic effect of the one or more therapeutic agent(s). in other embodiments, the present invention relates to methods of augmenting the therapeutic effect in a patient of one or more therapeutic agent(s) by administering another therapeutic agent to the patient.

In certain embodiments, the invention is directed in part to synergistic combinations of one or more therapeutic agent(s) in an amount sufficient to render a therapeutic effect together with the remaining therapeutic agent(s). For example, in certain embodiments a therapeutic effect is attained which is at least about 2 (or at least about 4, 6, 8, or 10) times greater than that obtained with the dose of the one or more therapeutic agent(s) alone. In certain embodiments, the synergistic combination provides a therapeutic effect which is up to about 20, 30 or 40 times greater than that obtained with the dose of the one or more therapeutic agent(s) alone, in such embodiments, the synergistic combinations display what is referred to herein as an "apparent one-way synergy", meaning that the dose of the remaining therapeutic agent(s) synergistica!iy potentiates the effect of the one or more therapeutic agent(s), but the dose of the one or more therapeutic agent(s) does not appear to significantly potentiate the effect of the remaining therapeutic agent(s).

In certain embodiments, the combination of active agents exhibits two-way synergism, meaning that the second therapeutic agent potentiates the effect of the first therapeutic agent, and the first therapeutic agent potentiates the effect of the second therapeutic agent. Thus, other embodiments of the invention relate to combinations of a second therapeutic agent and a first therapeutic agent where the dose of each drug is reduced due to the synergism between the drugs, and the therapeutic effect derived from the combination of drugs in reduced doses is enhanced. The two-way synergism is not always readily apparent in actual dosages due to the potency ratio of the first therapeutic agent to the second therapeutic agent. For instance, two-way synergism can be difficult to defect when one therapeutic agent displays much greater therapeutic potency relative to the other therapeutic agent.

The synergistic effects of combination therapy may be evaluated by biological activity assays. For example, the therapeutic agents are mixed at molar ratios designed to give approximately equipotent therapeutic effects based on the EC90 values. Then, three different molar ratios are used for each combination to allow for variability in the estimates of relative potency. These molar ratios are maintained throughout the dilution series. The corresponding monotherapies are also evaluated in parallel to the combination treatments using the standard primary assay format. A comparison of the therapeutic effect of the combination treatment to the therapeutic effect of the monotherapy gives a measure of the synergistic effect. Further details on the design of combination analyses can be found in B E Korba (1996) Antiviral Res. 29:49. Analysis of synergism, additivity, or antagonism can be determined by analysis of the aforementioned data using the CalcuSyn™ program (Biosoft, Inc.). This program evaluates drug interactions by use of the widely accepted method of Chou and Talalay combined with a statistically evaluation using the Monte Carlo statistical package. The data are displayed in several different formats including median- effect and dose-effects plots, isobolograms, and combination index [Ci] plots with standard deviations. For the latter analysis, a CI greater than 1.0 indicates antagonism and a CI less than 1.0 indicates synergism.

Compositions of the invention present the opportunity for obtaining relief from moderate to severe cases of disease. Due to the synergistic or additive or augmented effects provided by the inventive combination of the first and second therapeutic agent, it may be possible to use reduced dosages of each of therapeutic agent. By using lesser amounts of drugs, the side effects associated with each may be reduced in number and degree. Moreover, the inventive combinations avoid side effects to which some patients are particularly sensitive

Exemplary Combinations

In certain embodiments, the invention relates to a method of treating a metabolic disease, comprising the step of chronically co-administering to a mammal in need thereof a therapeutically effective amount of a first compound, and therapeutically effective amount of a second compound, wherein the first compound inhibits FAP, and the second compound inhibits DPP4.

In certain embodiments, the invention relates to any one of the aforementioned methods, wherein the metabolic disease is selected from the group consisting of diabetes, obesity, overweight, shortness of breath, gallbladder disease, hypertension, elevated blood cholesterol levels, cancer (e.g., endometrial, breast, prostate, colon), osteoarthritis, other orthopedic problems, reflux esophagitis (heartburn), snoring, menstrual irregularities, infertility, heart trouble, dyslipidemia, coronary heart disease, stroke, hyperinsulinernia, depression, anxiety, gout, fatty liver disease, insulin resistance, pre-diabetes, metabolic syndrome, beta-cell dysfunction, sleep apnea, obstructive sleep apnea, hypopnea, and visceral adiposity. In certain embodiments, the invention relates to any one of the aforementioned methods, wherein the metabolic disease is type 2 diabetes, hyperinsulinernia, or insulin resistance. In certain embodiments, the invention relates to any one of the aforementioned methods, wherein the metabolic disease is type 2 diabetes.

The subject muitimediator inhibitors can be used alone or in combination with one or more: anti-diabetics; anti-hypergiycemic agents; hypolipidemic/lipid lowering agents; anti- obesity agents; anti-hypertensive agents appetite suppressants; insulin secretagogues, insulin sensitizers, giucokinase activators, glucocorticoid antagonist, fructose 1,6-bis phosphatase inhibitors, AMP kinase activators, modulators of the incretin pathway such as incretin secretagogues such as GPR119 or GPR40 agonists, incretin mimics such as Byetta, and incretin potentiators, bile acid sequestrants or bile acid receptor agonists such as TGR5 agonists, dopamine receptor agonists such as Cycloset, aldose reductase inhibitors PPAR.gamma. agonists, PPAR.aipha. agonists, PPAR.gamma. antagonists or agonists, PPAR.aipbaJ.gamrna, dual agonists, ll-.beta.-HSD-l inhibitors, dipeptidyl peptidase IV (DPP4) inhibitors other than saxagliptin, SGLT2 inhibitors other than dapagliflozin, glucagon-like peptide-l (GLP-1), GLP-1 agonists, and PTP-1B inhibitors. Also weight loss agents acting to decreasing food intake such as sibutrimine, CB1 antagonists, 5HT2C agonists, MCH 1 antagonists, and agents which decrease nutrient absorption (such as lipase inhibitors (Orlistat)), and agents which increase energy expenditure such as thyromimetics, or slow Gl motility such as amylin mimetics or ghrelin antagonists.

Examples of suitable anti-diabetic agents for use in combination with the formulations of the present invention include, but are not limited to, alpha glucosidase inhibitors (acarbose or miglitol), insulins (including insulin secretagogues or insulin sensitizers), megiitinides (repaglinide), sulfonylureas (glimepiride, glyburide, gliclazide, chlorpropamide and glipizide), biguanide/giyburide combinations (Glucovance), thiazolidinediones (e.g., troglitazone, rosigiitazone and pioglitazone), PPAR-alpha agonists, PPAR-gamma agonists, PPAR alpha/gamma dual agonists, glycogen phosphorylase inhibitors, inhibitors of fatty acid binding protein (aP2), GPR-119 modulators, GPR 40 modulators, giucokinase inhibitors, glucagon-like peptide-l (GLP-1) and other agonists of the GLP-1 receptor, SGLT2 inhibitors other than dapagliflozin, and dipeptidyl peptidase IV (DPP4) inhibitors other than saxagliptin.

Other suitable thiazolidinediones include, but are not limited to, MCC-555 (disclosed in U.S. Pat. No. 5,594,016, Mitsubishi), faraglitazar (Gi-262570, Glaxo-Wellcome), englitazone (CP- 68722, Pfizer) or darglitazone (CP-86325, Pfizer; isaglitazone, MIT/Johnson& Johnson), reglitazar OTT-5Q1, (JPNT/Pharmacia & Upjohn), rivoglitazone (R-1197Q2, Sankyo/WL), liraglutide (NN-2344, Dr. Reddy/NN), and (Z)-l,4-bis-4-[(3,5-dioxo-l,2,4-oxadiazolidin-2-yl- methyl)]phenoxybut-2-e- ne (YM-440, Yamanouchi).

Examples of PPAR-a!pha agonists, PPAR-gamma agonists and PPAR alpha/gamma dual agonists include, but are not limited to, muraglitazar, peliglitazar, tesagiitazar AR-H039242 (Astra/Zeneca), GW-501516 (Glaxo-Wellcome), KRP297 (Kyorin Merck), as well as those disclosed by Murakami et al, "A Novel Insulin Sensitizer Acts As a Coligand for Peroxisome Proliferation-Activated Receptor Alpha (PPAR alpha) and PPAR gamma. Effect on PPAR alpha Activation on Abnormal Lipid Metabolism in Liver of Zucker Fatty Rats", Diabetes 47, 1841-1847 (1998); WO 01/21602 and in U.S. Pat. No. 6,414,002 and U.S. Pat. No. 6,653,314, the disclosures of which are incorporated herein by reference in their entireties, employing dosages as set out therein.

Suitable aP2 inhibitors inciude, but are not limited to, those disclosed in U.S. application Ser. No. 09/391,053, filed Sep. 7, 1999, and in U.S. Pat. No. 6,548,529, the disclosures of which are incorporated herein by reference in their entireties, employing dosages as set out therein.

Suitable SGLT2 inhibitors contemplated by the present invention's bilayer coated tablet and combination therapy with the present invention's bilayer tablet include sergliflozin, remogiiflozin, remogiif!ozin etabonate, canaglifiozin, Bi-10773 and B!-44847, ASP-1941, R- 7201, LX-4211, YM-543, AVE 2268, TS-033 or SGL-0100, and the compounds disclosed in U.S. Pat. No. 7,589,193, WO2007007628, EP2009010, WO200903596, US2009030198, U.S. Pat. No. 7,288,528 and US 2007/0197623, herein incorporated by reference in their entirety for any purpose.

Suitable meglitinides include nateglinide (Novartis) or KAD1229 (PF/Kissei).

Examples of suitable anti-hypergiycemic agents for use in combination with the formulations of the present invention include, but are not limited to, glucagon-like peptide- 1 (GLP-1) such as GLP-l(l-36) amide, GLP-l{7-36) amide, GLP-l{7-37) (as disclosed in U.S. Pat. No. 5,614,492, incorporated herein by reference in its entirety), as well as exenatide (Amylin/Liily), LY-315902 (Lilly), MK-0431 (Merck), liraglutide (NovoNordisk), ZP-10 (Zealand Pharmaceuticals A/S), CJC-1131 (Conjuchem Inc), and the compounds disclosed in WO 03/033671, incorporated herein by reference in its entirety.

Examples of suitable hypolipidemic/lipid lowering agents for use in combination with the formulations of the present invention include one or more MTP inhibitors, HMG CoA reductase inhibitors, squalene synthetase inhibitors, fibric acid derivatives, ACAT inhibitors, lipoxygenase inhibitors, cholesterol absorption inhibitors, ileal Na bile acid co-transporter inhibitors, up-regulators of LDL receptor activity, bile acid sequestrants, cholesterol ester transfer protein (e.g., CETP inhibitors, such as torcetrapib (CP-529414, Pfizer) and JTT-705 (Akros Pharma)), PPAR agonists (as described above) and/or nicotinic acid and derivatives thereof. The hypolipidemic agent can be an up-reguiator of LD2 receptor activity, such as l(3H)-isobenzofuranone,3-(13-hydroxy-10-oxotetradecyl)-5_/7-dimethoxy-(MD--700, Taisho Pharmaceutical Co. Ltd) and cho!estan-3-ol,4-(2-propenyi)-(3a,4a,5a)-(LY295427, Eli Lilly). Hypolipidemic agents include for example pravastatin, iovastatin, simvastatin, atorvastatin, fluvastatin, cerivastatin, atavastatin and rosuvastatin (ZD-4522).

Examples of MTP inhibitors that can be employed as described above include, but are not limited to, those disclosed in U.S. Pat. No, 5,595,872, U.S. Pat. No. 5,739,135, U.S. Pat, No. 5,712,279, U.S. Pat. No. 5,760,246, U.S. Pat. No. 5,827,875, U.S. Pat. No. 5,885,983 and U.S. Pat. No. 5,962,440, all of which are incorporated herein by reference in their entireties.

Examples of HMG CoA reductase inhibitors that can be employed in combination with the formulations of the invention include, but are not limited to, mevastatin and related compounds, as disclosed in U.S. Pat. No. 3,983,140, Iovastatin (mevinolin) and related compounds, as disclosed in U.S. Pat. No. 4,231,938, pravastatin and related compounds, such as disclosed in U.S. Pat. No. 4,346,227, simvastatin and related compounds, as disclosed in U.S. Pat. Nos. 4,448,784 and 4,450,171. Other suitable HMG CoA reductase inhibitors that can be employed herein include, but are not limited to, fluvastatin, disclosed in U.S. Pat. No. 5,354,772, cerivastatin, as disclosed in U.S. Pat. Nos. 5,006,530 and 5,177,080, atorvastatin, as disclosed in U.S. Pat. Nos. 4,681,893, 5,273,995, 5,385,929 and 5,686,104, atavastatin (Nissan/Sankyo's nisvastatin (NK-104)), as disclosed in U.S. Pat. No. 5,011,930, rosuvastatin (Shionogi-Astra/Zeneca (ZD-4522)), as disclosed in U.S. Pat. No. 5,260,440, and related statin compounds disclosed in U.S. Pat. No. 5,753,675, pyrazoie analogs of mevalonolactone derivatives, as disclosed in U.S. Pat. No. 4,613,610, indene analogs of mevalonolactone derivatives, as disclosed in PCT application WO 86/03488, 60[2-(substituted-pyrrol-l-yl)-alkyl)pyran-2-ones and derivatives thereof, as disclosed in U.S. Pat. No. 4,647,576, Searle's SC-45355 (a 3-substituted pentanedioic acid derivative) dichloroacetate, imidazole analogs of mevalonolactone, as disclosed in PCT application WO 86/07054, 3-carboxy-2-hydroxy-propane-phosphonic acid derivatives, as disclosed in French Patent No. 2,596,393, 2,3-disubstituted pyrrole, furan and thiophene derivatives, as disclosed in European Patent Application No. 0221025, naphthyl analogs of mevalonolactone, as disclosed in U.S. Pat. No. 4,686,237, octahydronaphthalenes, such as disclosed in U.S. Pat. No. 4,499,289, keto analogs of mevinolin (Iovastatin), as disclosed in European Patent Application No. 0142146 A2, and quinoiine and pyridine derivatives, as disclosed in U.S. Pat. Nos. 5,506,219 and 5,691,322. All of the cited references are incorporated herein by reference in their entireties, in addition, phosphinic acid compounds useful in inhibiting HMG CoA reductase, such as those disclosed in GB 2205837, are suitable for use in combination with the formulations of the present invention.

Examples of squalene synthetase inhibitors suitable for use herein include, but are not limited to, . alpha. -phosphono-suifonates disclosed in U.S. Pat. No. 5,712,396, those disclosed by Biiler et al., J. Med. Chem., 1988, Vol. 31, No. 10, pp. 1869-1871, including isoprenoid (phosphinyl-methyl)phosphonates, as well as other known squalene synthetase inhibitors, for example, as disclosed in U.S. Pat. !Mos. 4,871,721 and 4,924,024 and in Biiler, S. A., Neuenschwander, K., Ponpipom, M. M., and Poulter, C. D., Current Pharmaceutical Design, 2, 1-40 (1996). Other squalene synthetase inhibitors suitable for use herein include the terpenoid pyrophosphates disclosed by P. Ortiz de Monteliano et ai, J. Med. Chem., 1977, 20, 243-249; the farnesyi diphosphate analog A and presquaiene pyrophosphate (PSQ-PP) analogs as disclosed by Corey and Volante, J. Am. Chem. Soc, 1976, 98, 1291- 1293; phosphinylphosphonates reported by McClard, R. W. et al, J.A.C.S., 1987, 109, 5544; and cyciopropanes reported by Capson, T. L, PhD dissertation, June 1987, Dept. Med. Chem. U of Utah, Abstract, Table of Contents, pp 16, 17, 40-43, 48-51, Summary. AH of the cited references are incorporated herein by reference in their entireties.

Examples of fibric acid derivatives that can be employed in combination the formulations of the invention include, but are not limited to, fenofibrate, gemfibrozil, clofibrate, bezafibrate, ciprofibrate, ciinofibrate and the like, probucol, and related compounds, as disclosed in U.S. Pat. Mo. 3,674,836, bile acid sequestrants, such as cholestyramine, colestipol and DEAE-Sephadex (Sechoiex), Policexide), as well as lipostabii (Rhone-Poulenc), Eisai E-5050 (an M-substitufed ethanolamine derivative), imanixi! (HOE-402), tetrahydrolipstatin (THL), istigmastanylphos-phorylchoiine fSPC, Roche), aminocyclodextrin (Tanabe Seiyoku), Ajinomoto AJ-814 (azulene derivative), meiinamide (Sumitomo), Sandoz 58-035, American Cyanamid CL-277,082 and CL-283,546 (disubstituted urea derivatives), nicotinic acid, acipimox, acifran, neomycin, p-aminosalicyiic acid, aspirin, poly(diallylmethylamine) derivatives, such as disclosed in U.S. Pat. No. 4,759.923, quaternary amine poly(diailyidimethylammonium chloride) and ionenes, such as disclosed in U.S. Pat. No. 4,027,009, and other known serum cholesterol lowering agents, in one embodiment, the fibric acid derivative is probucol or gemfibrozil. All of the cited references are incorporated herein by reference in their entireties.

Examples of ACAT inhibitors that can be employed in combination with the formulations of the invention include, but are not limited to, those disclosed in Drugs of the Future 24, 9-15 (1999), (Avasimibe); "The ACAT inhibitor, Cl-1011 is effective in the prevention and regression of aortic fatty streak area in hamsters", Nicolosi et ai, Atherosclerosis (Shannon, irei). (1998), 137(1), 77-85; "The pharmacological profile of FCE 27677: a novel ACAT inhibitor with potent hypolipidemic activity mediated by selective suppression of the hepatic secretion of ΑροΒΙΟΟ-containing lipoprotein", Ghiselii, Giancarlo, Cardiovasc. Drug Rev, (1998), 16(1), 16-30; "RP 73163: a bioavaiiable alkylsulfinyl-diphenylimidazole ACAT inhibitor", Smith, C, et al, Bioorg. Med. Chem. Lett. (1996), 6(1), 47-50; "ACAT inhibitors: physiologic mechanisms for hypolipidemic and anti-atherosclerotic activities in experimental animals", Krause ef al, Editor(s): Ruffolo, Robert R., Jr.; Hollinger, Mannfred A., Inflammation: Mediators Pathways (1995), 173-98, Publisher: CRC, Boca Raton, Fla.; "ACAT inhibitors: potential anti-atherosclerotic agents", Sliskovic et al, Curr. Med. Chem. (1994), 1(3), 204-25; "Inhibitors of acyl-CoA: cholesterol O-acyl transferase (ACAT) as hypocho!esterolemic agents. The first water-soluble ACAT inhibitor with lipid-regulating activity. Inhibitors of acyl-CoA:cholesterol acyltransferase (ACAT). Development of a series of substituted N-phenyl-N'-[(l-phenylcyclopentyl)iTiethyl]ureas with enhanced hypocholesterolemic activity", Stout et al, Chemfracfs: Org. Chem. (1995), 8(6), 359-62, or TS-962 (Taisho Pharmaceutical Co. Ltd). All of the cited references are incorporated herein by reference in their entireties.

Examples of suitable cholesterol absorption inhibitors for use in combination with the formulations of the invention include, but are not limited to, SCH48461 (Schering-Plough), as well as those disclosed in Atherosclerosis 115, 45-63 (1995) and J. Med. Chem. 41, 973 (1998), incorporated herein by reference in its entirety.

Examples of suitable ileal Na.sup.+/bile acid co-transporter inhibitors for use in combination with the formulations of the invention include, but are not limited to, compounds as disclosed in Drugs of the Future, 24, 425-430 (1999), incorporated herein by reference in its entirety.

Examples of lipoxygenase inhibitors that can be employed in combination with the formulations of the invention include, but are not limited to, 15-lipoxygenase (15-LO) inhibitors, such as benzimidazoie derivatives, as disclosed in WO 97/12615, 15-LO inhibitors, as disclosed in WO 97/12613, isothiazolones, as disclosed in WO 96/38144, and 15-LO inhibitors, as disclosed by Sendobry et al "Attenuation of diet-induced atherosclerosis in rabbits with a highly selective 15-lipoxygenase inhibitor lacking significant antioxidant properties", Brit. J. Pharmacology (1997) 120, 1199-1206, and Cornicelli et al., "15-Lipoxygenase and its inhibition: A Novel Therapeutic Target for Vascular Disease", Current Pharmaceutical Design, 1999, 5, 11-20. Ail of the cited references are incorporated herein by reference in their entireties.

Examples of suitable anti-hypertensive agents for use in combination with the formulations of the present invention include, but are not limited to, beta adrenergic blockers, calcium channel blockers (L-type and T-type; e.g. diltiazem, verapamil, nifedipine, amlodipine and mybefradil), diuretics (e.g., chlorothiazide, hydrochlorothiazide, fiumethiazide, hydroflumethiazide, bendroflumethiazide, methylchiorothiazide, trichloromethiazide, polythiazide, benzthiazide, ethacrynic acid tricryrsafen, chlorthalidone, furosemide, musolimine, bumetanide, triamtrenene, amiloride, spironolactone), renin inhibitors. ACE inhibitors (e.g., captopril, zofenopril, fosinopnl, enalapril, ceranopril, cilazopnl, delapril, pentopril, quinapril, ramipril, !isinopril), AT-1 receptor antagonists (e.g., losartan, irbesartan, valsartan), ET receptor antagonists (e.g., sitaxsentan, atrsentan and compounds disclosed in U.S. Pat. Nos. 5,612,359 and 6,043,265), Dual ET/AM antagonist (e.g., compounds disclosed in WO 00/01389), neutral endopeptidase (NEP) inhibitors, vasopepsidase inhibitors (dual NEP-ACE inhibitors) (e.g., omapatrilat and gemopatrilat), and nitrates. All of the cited references are incorporated herein by reference in their entireties.

Examples of suitable anti-obesity agents for use in combination with the formulations of the present invention include, but are not limited to, beta 3 adrenergic agonists, lipase inhibitors, serotonin (and dopamine) reuptake inhibitors, thyroid receptor beta drugs, 5HT2C agonists, (such as Arena APD-356); MCHR1 antagonists, such as Synaptic SNAP-7941 and Takeda T-226926, melanocortin receptor (MC4R) agonists, melanin-concentrating hormone receptor (SvlCHR) antagonists (such as Synaptic SNAP-7941 and Takeda T-226926), galanin receptor modulators, orexin antagonists, CCK agonists, NPY1 or NPY5 antagonist, NPY2 and NPY4 modulators, corticotropin releasing factor agonists, histamine receptor-3 (H3) modulators, 11-beta-HSD-l inhibitors, adinopectin receptor modulators, monoamine reuptake inhibitors or releasing agents, ciliary neurotrophic factors (CiMTF, such as AXQKINE.RTM. by Regeneron), BDIMF (brain-derived neurotrophic factor), leptin and leptin receptor modulators, cannabinoid-1 receptor antagonists (such as SR-141716 (Sanofi) or SLV-319 (Solvay)), and anorectic agents.

Beta 3 adrenergic agonists that can be optionally employed in combination with formulations of the present invention include, but are not limited to, AJ9677 (Takeda/Dainippon), L750355 (Merck), CP331648 (Pfizer,) or other known beta 3 agonists, as disclosed in U.S. Pat. Nos. 5,541,204, 5,770,615, 5,491,134, 5,776,983 and 5,488,064, ail of which are incorporated herein by reference in their entireties.

Examples of lipase inhibitors that can be employed in combination with formulations of the present invention include, but are not limited to, orlistat and ATL-962 (Aiizyme).

Serotonin (and dopamine) reuptake inhibitors (or serotonin receptor agonists) that can be employed in combination with the formulations of the present invention inciude, but are not limited to, BVT-933 (Biovitrum), sibutramine, topiramate (Johnson & Johnson) and axokine (Regeneron).

Examples of thyroid receptor beta compounds that can be employed in combination with formulations of the present invention include, but are not limited to, thyroid receptor iigands, such as those disclosed in WO 97/21993 (U. Cai SF), WO 99/00353 (KaroBio) and WO 00/039077 (KaroBio), incorporated herein by reference it their entireties.

Examples of monoamine reuptake inhibitors that can be employed in combination with the formulations of the present invention include, but are not limited to, fenfluramine, dexfenfluramine, fluvoxamine, fluoxetine, paroxetine, sertraline, chlorphentermine, cloforex, clortermine, piciiorex, sibutramine, dexamphetamine, phentermine, phenylpropanolamine and mazindo!.

Anorectic agents that can be employed in combination with the formulations of the present invention include, but are not limited to, topiramafe (Johnson & Johnson), dexamphetamine, phentermine, phenylpropanolamine and mazindoi.

Exemplary Uses

Another aspect of the present invention relates to the use of any one of the compounds disclosed herein in the manufacture of a medicament for the treatment of a metabolic disease.

Another aspect of the present invention relates to the use of a prodrug of any one of the compounds disclosed herein in the manufacture of a medicament for the treatment of a metabolic disease.

Exemplary Packaged Pharmaceuticals

Another aspect of the present invention relates to a packaged pharmaceutical, comprising any one of the compounds disclosed herein formulated in a pharmaceutically acceptable excipient, in association with instructions (written and/or pictorial) describing the recommended dosage and/or administration of the formulation to a patient.

Another aspect of the present invention relates to a packaged pharmaceutical, comprising a prodrug of any of the compounds disclosed herein formulated in a pharmaceutically acceptable excipient, in association with instructions (written and/or pictorial) describing the recommended dosage and/or administration of the formulation to a patient. Examples

Claims

We claim :

1. A pharmaceutical preparation, comprising a muitimediator inhibitor agent that both inhibits DPP4 enzymatic activity with a Ki of 100 n or less and inhibits FAP enzymatic activity with a Ki of 100 n or less, which muitimediator is orally active and is formulated for oral administration at a dosing frequency of 24 hours or greater; and a pharmaceutically acceptable excipient.

2. The pharmaceutical preparation of claim 1, wherein the muitimediator inhibits DPP4 enzymatic activity with a K₀ff rate less than 1 x 10^~4/sec, and inhibits FAP enzymatic activity with a K_Qff rate less than 1 x lQ^~4/sec.

3. The pharmaceutical preparation of claim 1 or 2, wherein the muitimediator has an IC50 for inhibition of DPP-8 and DPP-9 that is at least one order of magnitude greater than the IC50 for inhibition of DPP4 and FAP,

4. The pharmaceutical preparation of claim 3, wherein the order of magnitude is 1 microM or greater,

5. The pharmaceutical preparation of any one of the preceding claims, wherein the muitimediator is represented in the general formula:

wherein

R¹ represents a naturally or non-naturally occurring amino acid side chain;

R² represents a moiety including two or more overlapping p-orbitals with delocaiized electrons, which forms a conjugated system with the nitrogen to which R² is attached such that the nitrogen is a tertiary amine which maintains an electron lone pair under conditions in which the muitimediator interacts with DPP4;

R⁵ represents a hydrogen, a lower a!kyl, or a -lower alkyi-haiogen (such as to form a ha!o-a!kyl ketone, i.e., -CH3-CI);

R⁷ represents, for each occurrence, hydrogen, or a substituted or unsubstituted lower aikyl, lower alkenyl, aryl, araikyl, cycioaikyl, cycioaikenyl, or heterocycle;

R⁵⁰ represents O or S; R⁵ⁱ represents N₃, SH₂, NH₂, H0₂ or OR⁷;

R⁵² represents hydrogen, a lower a!kyi, amine, OR⁷, or a pharmaceutically acceptable salt, or R^bl and R^b2 taken together with the phosphorous atom to which they are attached complete a heterocyclic ring having from 5 to 8 atoms in the ring structure;

Z represents a 4-12 membered heterocycie;

W represents a functional group which reacts with an active site residue of DPP4 and FAP, as for example,— CN,— CH=NR⁵,

o o R⁵⁰ O

— S— X¹ , — P— X¹ , P— R⁵² , — & , or— C— R⁵

j "~-γ2

X represents O or S;

X^I represents a halogen; and

The pharmaceutical preparation of claim 5, wherein the ring Z is represented by

R³ is absent or represents from 1 to 3 ring substifuents, each of which can independently be a halogen, a lower a!kyl, a lower aikenyl, a lower alkynyl, a carbonyl (such as a carboxyl, an ester, a formate, or a ketone), a thiocarbonyi (such as a thioester, a thioacetate, or a thioformate), an amino, an acyiamino, an amido, a cyano, a nitro, an azido, a sulfate, a sulfonate, a sulfonamide), — {CH2)_m—^■ R⁷,—^■ {CH₂)_m-0-R⁷,-(CH₂)_m-S-R⁷;

n represents 1, 2, 3 or 4; and

m represents zero or an integer from 1 to 8.

The pharmaceutical preparation of claim 6, wherein the ring Z is represented by:

8. The pharmaceutical preparation of any one of the preceding claims, wherein R² is an aromatic moiety which forms the conjugated system with the nitrogen to which R² is attached.

9. The pharmaceutical preparation of claim 5, wherein the muitimediator is represented in the general formula:

wherein

R³ is absent or represents from 1 to 3 ring substituents, each of which can independently be a halogen, a lower alkyi, a lower alkenyi, a lower aikynyi, a carbonyl (such as a carboxyl, an ester, a formate, or a ketone), a thiocarbonyl (such as a thioester, a thioacetate, or a thioformate), an amino, an acylamino, an amido, a cyano, a nitro, an azido, a sulfate, a sulfonate, a suifonamido, — (Chkjm— ⁷,— iCH₂)_m— O— R⁷,— (C^— S—R⁷;

n represents 1, 2, 3 or 4; and

m represents zero or an integer from 1 to 8.

10. The pharmaceutical preparation of any one of the preceding claims, wherein R¹ is a hydrogen or a small hydrophobic group.

11. The pharmaceutical preparation of any one of the preceding claims, wherein aromatic moiety is a hydroxylated benzene moiety, substituted with from 1 to 5 hydroxyl groups.

12. The pharmaceutical preparation of claim 11, wherein the hydroxylated benzene moiety is substituted with 2 hydroxyl groups.

13. The pharmaceutical preparation of any one of the preceding claims, wherein aromatic moiety is a haiogenated benzene moiety, substituted with from 1 to 5 halogen groups.

14. The pharmaceutical preparation of claim 13, wherein the haiogenated benzene moiety is substituted with 2 halogen groups.

15. The pharmaceutical preparation of claim 13 or 14, where the halogen groups are CI, Br or F.

16. The pharmaceutical preparation of claim 9, wherein the multimediator is represented in the general formula:

wherein

R¹ is hydrogen or a lower alkyl;

R^3a and R^3b are each hydrogen, or R^3a is hydrogen and R^3b is -OH, or R^3a and R^3b each independently are a halogen; and

R⁵ represents 1, 2 or 3 hydroxyl groups, or R⁶ represents 1, 2 or 3 halogens.

17. The pharmaceutical preparation of claim 16, wherein the halogens are F.

18. The pharmaceutical preparation of any one of the preceding claims, wherein the multimediator is formulated in a single oral dosage form containing less than 500 mg of the multimediator.

19. The pharmaceutical preparation of any one of the preceding claims, further including one or more additional active ingredients selected from the group consisting of anti-diabetic agents, anti-hyperglycemic agents, hypolipidemic/lipid lowering agents, anti-obesity agents, anti-hypertensive agents and appetite suppressants.

20. A compound represented in the general formula:

wherein

R¹ represents a naturally or non-naturally occurring amino acid side chain;

R⁵ represents a hydrogen, a lower alkyl, or a -lower a!ky!-haiogen (such as to form a halo-alky! ketone, i.e., -CH3-CI);

R⁵⁰ represents O or S;

R⁵¹ represents N₃, SH₂, N H₂, N0₂ or OR⁷;

Z represents a 4-12 rnembered heterocycle;

W represents a functional group which reacts with an active site residue of DPP4 and FAP, as for example,— CM,— -CH=N R⁵,

X represents O or S;

X^I represents a halogen; and

Yi and Y₂ can independently or together be OH, or a group capable of being hydro!yzed to a hydroxyl group, including cyclic derivatives where Yi and Y₂ are connected via a ring having from 5 to 8 atoms in the ring structure (such as pinacol or the like).

21. A method for reducing a blood glucose level of a subject having an elevated blood glucose level or preventing or delaying the onset of an elevated blood glucose level in a subject at risk for developing an elevated glucose level, comprising administering to the subject an effective amount of a pharmaceutical preparation of any one of claims 1-19.

22. A method for improving insulin sensitivity in a subject with insulin resistance or delaying the onset of insulin resistance in a subject at risk for developing insulin resistance in a subject, comprising administering to the subject an effective amount of a pharmaceutical preparation of any one of claims 1-19.

23. A method for improving glucose tolerance in a subject with impaired glucose tolerance, comprising administering to the subject an effective amount of a pharmaceutical preparation of any one of claims 1-19.

24. A method of treating at least one metabolic disorder in a subject or preventing or delaying the onset of at least one metabolic disorder in a subject, comprising administering to the subject having a metabolic disorder an effective amount of a pharmaceutical preparation of any one of claims 1-19, wherein the metabolic disorder is optionally selected from pre-diabetes, diabetes, metabolic syndrome, obesity, diabetic dys!ipidemia, hyperlipdemia, hypertension, hypertriglyceridemia, hyperfattyacidemia, hypercholesterolemia, and hyperinsulinemia.

25. A method for treating fatty liver disease in a subject, optionally selected from nonalcoholic fatty liver disease (NAFLD), alcoholic fatty liver disease, alcoholic steatohepatitis, and non-alcoholic steatohepatitis (NASH), comprising administering to the subject having a fatty liver disease an effective amount of a pharmaceutical preparation of any one of claims 1-19.

26. A method of reducing liver triglycerides, comprising administering to a subject an effective amount of a pharmaceutical preparation of any one of claims 1-19.

27. A method of treating a patient having a cardiovascular and/or renal microvascular disease, the method comprising administering to the patient an effective amount of a pharmaceutical preparation of any one of claims 1-19.

28. The method of claim 26, wherein the treatment is both cardioprotective and renoprotective.

29. The method of claim 26, wherein the treatment protects against, delays the occurrence of, delays the progression of and/or reduces the risk of at least one disease selected from the group consisting of CV morbidity, premature CV mortality, renai morbidity and premature renai mortality.

30. The method of claim 26, wherein the treatment (a) protects against, reduces the risk of and/or delays the occurrence of: a cardio- or cerebrovascular disease or event selected from the group consisting of cardiovascular (CV) death selected from the group consisting of fatal stroke, fatal myocardial infarction, fatal heart failure, cardiogenic shock and sudden death, non-fatal stroke and non-fatal myocardial infarction (M l) (with or without silent M l), and, optionally, hospitalization, wherein said hospitalization is for unstable angina pectoris, stable angina pectoris, transient ischemic attack, coronary revascularization procedures, peripheral revascularization or congestive heart failure, and/or (b) protects against, reduces the risk of, delays the progression of and/or delays the occurrence of a renal microvascular disease selected from the group consisting of: abuminuria, chronic kidney disease (CKD), renal impairment, renal death, end-stage renal disease and loss in estimated glomerular filtration rate.

31. A method of slowing the progression of, delaying the onset of or treating a metabolic disorder or disease, comprising administering to a subject an effective amount of a pharmaceutical preparation of any one of claims 1-19.

32. The method of claim 31, wherein the metabolic disease or disorder is selected from the group consisting of type 1 diabetes meilitus, type 2 diabetes mellitus, impaired glucose tolerance (IGT), impaired fasting blood glucose (IFG), hyperglycemia, postprandial hyperglycemia, postabsorptive hyperglycemia, latent autoimmune diabetes in adults (LADA), overweight, obesity, dyslipidemia, hyperlipidemia, hypercholesterolemia, hypertriglyceridemia, hyperNEFA-emia, postprandial iipemia, hypertension, atherosclerosis, endothelial dysfunction, osteoporosis, chronic systemic inflammation, non alcoholic fatty liver disease (NAFLD), retinopathy, neuropathy, nephropathy, nephrotic syndrome, polycystic ovarian syndrome, and/or metabolic syndrome; improving and/or maintaining giycemic control and/or for reducing of fasting plasma glucose, of postprandial plasma glucose, of postabsorptive plasma glucose and/or of glycosylated hemoglobin HbAlc; slowing, delaying or reversing progression from prediabetes, impaired glucose tolerance (IGT), impaired fasting blood glucose (IFG), insulin resistance and/or from metabolic syndrome to type 2 diabetes mellitus; reducing the risk of, slowing the progression of, delaying the onset of or treating complications of diabetes mellitus selected from the group consisting of micro- and macrovascular diseases, nephropathy, micro- or macroalbuminuria, proteinuria, nephrotic syndrome, retinopathy, cataracts, neuropathy, learning or memory impairment, neurodegenerative or cognitive disorders, cardio- or cerebrovascular diseases, tissue ischaemia, diabetic foot or ulcer, atherosclerosis, hypertension, endothelial dysfunction, myocardial infarction, acute coronary syndrome, unstable angina pectoris, stable angina pectoris, peripheral arterial occlusive disease, cardiomyopathy, heart failure, heart rhythm disorders, vascular restenosis, and stroke; reducing body weight and/or body fat and/or liver fat and/or intra- myocellular fat or facilitating a reduction in body weight and/or body fat and/or liver fat and/or intra-myocelluiar fat; slowing, delaying the onset of or treating the degeneration of pancreatic beta cells and/or the decline of the functionality of pancreatic beta cells and/or for improving, preserving and/or restoring the functionality of pancreatic beta ceils and/or stimulating and/or restoring or protecting the functionality of pancreatic insulin secretion; slowing, delaying the onset of or treating non alcoholic fatty liver disease (NAFLD) including hepatic steatosis, non-alcoholic steatohepatitis (MASH) and/or liver fibrosis; slowing the progression of, delaying the onset of or treating type 2 diabetes with failure to conventional antidiabetic mono- or combination therapy; achieving a reduction in the dose of conventional antidiabetic medication required for adequate therapeutic effect; reducing the risk for adverse effects associated with conventional antidiabetic medication; and maintaining and/or improving the insulin sensitivity and/or for treating hyperinsulinemia and/or insulin resistance.

33. The method of claim 31, wherein the treatment protects against, delays the occurrence of, delays the progression of and/or reduces the risk of accelerated cognitive decline or impairment, dementia, and/or depressive, mood or anxiety disorders.

34. The method of claim 31, further comprising administering one or more other active agents selected from the group consisting of other antidiabetic substances, active substances that lower the blood sugar level, active substances that lower the lipid level in the blood, active substances that raise the HDL level in the blood, active substances that lower blood pressure, active substances that are indicated in the treatment of atherosclerosis or obesity, and/or active substances which are indicated in the treatment or prevention of major CV events and antiplatelet agents and/or anticoagulants.

35. A method for inducing satiety in a subject, comprising administering to the subject an effective amount of a pharmaceutical preparation of any one of claims 1-19. A method for reducing fat intake, bodyweight, and/or body fat in a subject, comprising administering to the subject an effective amount of a pharmaceutical preparation of any one of claims 1-19.