# Patent

## Title
Substituted amino acids as erythropoietin mimetics

## Abstract

This invention relates to a series of substitituted amino acids of Formula I




pharmaceutical compositions containing them and intermediates used in their manufacture. The compounds of the invention are small molecules which bind to the erythropoietin receptor and compete with the natural ligand for binding to this receptor.

## Background
This application is a divisional application of U.S. application Ser. No. 09/927,111, filed Aug. 10, 2001 now U.S. Pat. No. 6,750,369 to which no matter has been added. Such earlier application is hereby incorporated into the present application by reference and which is a divisional application of U.S. application Ser. No. 09/517,976, filed Mar. 3, 2000, which is now U.S. Pat. No. 6,310,078, which is a Continuation-in-part of U.S. application Ser. No. 09/294,785, filed Apr. 19, 1999 now abandoned, which claims benefit of U.S. Provisional Application 60/082,392, filed Apr. 20, 1998.


This invention relates to a series of small molecules which bind to the erythropoietin receptor and compete with the natural ligand for binding to said receptor. The invention includes pharmaceutical compositions containing these mimetics, their methods of production as well as intermediates used in their synthesis.
Erythropoietin (EPO) is a 34,000 dalton glycoprotein hormone which is produced in the mammalian kidney. Its primary role is stimulation of mitotic cell division and differentiation of erythrocyte precursor cells. As a result this hormone regulates the production of erythrocytes, the hemoglobin contained therein and the blood's ability to carry oxygen. The commercial product Epogen® is used in the treatment of anemia. This drug is produced by recombinant techniques and is formulated in aqueous isotonic sodium chloride/sodium citrate. Even though it has been used successfully in the treatment of anemia, it is a costly drug that is administered intravenously. This method of administration is both costly and inconvenient for the patient; therefore it would be desirable to find a EPO mimetic which has the potential for oral activity.
A small molecule EPO mimetic has advantages over the natural protein. The immune response associated with large peptides is unlikely to occur with small molecules. In addition, the variety of pharmaceutical formulations that may be used with small molecules are technically unfeasible for proteins. Thus the use of relatively inert formulations for small molecules is possible. The most important advantage of small molecules is their potential for oral activity. Such an agent would ease administration, cost less and facilitate patient compliance.
Although compounds which mimic EPO are useful in stimulating red blood cell synthesis, there are diseases where the overproduction of red blood cells is a problem. Erythroleukemia and polysythemia vera are examples of such diseases. Since EPO is an agent responsible for the maturation of red blood cell precursors, an antagonist of EPO would have utility treating either of those diseases.
SUMMARY OF THE INVENTION
The disclosed invention consists of a series of small molecules which demonstrate competitive binding with the natural ligand for the EPO receptor. As such these compounds are potentially useful in the treatment of diseases or conditions associated with this receptor. In addition, the invention contemplates methods of producing these compounds and intermediates used in their production.
The invention includes compounds of the Formula I:




wherein:

    
    
        R1 is the side chain of a natural or unnatural α-amino acids, where if said side chain contains a protectable group, that group may be protected with a member of the group consisting of succinyl, glutaryl, 3,3-dimethylglutaryl, C1-5alkyl, C1-5alkoxycarbonyl, acetyl, N-(9-fluorenylmethoxycarbonyl), trifluoroacetyl, omega-carboxyC1-5alkylcarbonyl, t-butoxycarbonyl, benzyl, benzyloxycarbonyl, 2-chlorobenzyloxycarbonyl, phenylsulfonyl, ureido, t-butyl, cinnamoyl, trityl, 4-methyltrityl, 1-(4,4-dimethyl-2,6-dioxocyclohexylidene)ethyl, tosyl, 4-methoxy-2,3,6-trimethylbenzenesulfonyl, phenylureido, and substituted phenylureido (where the phenyl substituents are phenoxy, halo, C1-5alkoxycarbonyl);
        R2 and R3 
        
            may be taken together to form a six-membered aromatic ring which is fused to the depicted ring, or
            are independently selected from the group consisting of hydrogen, C1-5 alkyl, C1-5 alkoxy, hydroxy, halo, trifluoromethyl, nitro, amino, phenyl, phenoxy, phenylC1-5alkyl, phenyl C1-5alkoxy,
            substituted phenyl (where the substituents are selected from C1-5alkyl, C1-5 alkoxy, hydroxy, halo, trifluoromethyl, nitro, cyano, and amino),
            substituted phenoxy (where the substituents are selected from C1-5 alkyl, C1-5 alkoxy, hydroxy, halo, trifluoromethyl, nitro, cyano, and amino),
            substituted phenylC1-5alkyl (where the substituents are selected from C1-5 alkyl, C1-5 alkoxy, hydroxy, halo, trifluoromethyl, nitro, cyano, and amino),
            substituted phenylC1-5alkoxy (where the substituents are selected from C1-5 alkyl, C1-5 alkoxy, hydroxy, halo, trifluoromethyl, nitro, cyano, and amino), and
            substituted amino (where the substituents are selected from one or more members of the group consisting of C1-5alkyl, halosubstitutedC1-5alkyl, C1-5alknyl, C1-5alkenyl, phenyl, phenylC1-5alkyl, C1-5alkylcarbonyl, halo substituted C1-5alkylcarbonyl, carboxyC1-5alkyl, C1-5alkoxyC1-5alkyl, cinnamoyl, naphthylcarbonyl, furylcarbonyl, pyridylcarbonyl, C1-5alkylsulfonyl, phenylcarbonyl, phenylC1-5alkylcarbonyl, phenylsulfonyl, phenylC1-5alkylsulfonyl substituted phenylcarbonyl, substituted phenylC1-5alkylcarbonyl, substituted phenylsulfonyl, substituted phenylC1-5alkylsulfonyl, substituted phenyl, and substituted phenylC1-5alkyl [where the aromatic phenyl, phenylC1-5alkyl, phenylcarbonyl, phenylC1-5alkylcarbonyl, phenylsulfonyl, and phenylC1-5alkylsulfonyl substitutents are independently selected from one to five members of the group consisting of C1-5alkyl, C1-5alkoxy, hydroxy, halogen, trifluoromethyl, nitro, cyano, and amino]);
        
        
        R4 and R5 
        
            may be taken together to form a six-membered aromatic ring which is fused to the depicted ring, or
            are independently selected from the group consisting of hydrogen, C1-5 alkyl, C1-5 alkoxy, hydroxy, halo, trifluoromethyl, nitro, amino, phenyl, phenoxy, phenylC1-5alkyl, phenyl C1-5alkoxy,
            substituted phenyl (where the substituents are selected from C1-5alkyl, C1-5 alkoxy, hydroxy, halo, trifluoromethyl, nitro, cyano, and amino),
            substituted phenoxy (where the substituents are selected from C1-5 alkyl, C1-5 alkoxy, hydroxy, halo, trifluoromethyl, nitro, cyano, and amino),
            substituted phenylC1-5alkyl (where the substituents are selected from C1-5 alkyl, C1-5 alkoxy, hydroxy, halo, trifluoromethyl, nitro, cyano, and amino),
            substituted phenylC1-5alkoxy (where the substituents are selected from C1-5 alkyl, C1-5 alkoxy, hydroxy, halo, trifluoromethyl, nitro, cyano, and amino), and
            substituted amino (where the substituents are selected from one or more members of the group consisting of C1-5alkyl, halosubstitutedC1-5alkyl, C1-5alknyl, C1-5alkenyl, phenyl, phenylC1-5alkyl, C1-5alkylcarbonyl, halo substituted C1-5alkylcarbonyl, carboxyC1-5alkyl, C1-5alkoxyC1-5alkyl, cinnamoyl, naphthylcarbonyl, furylcarbonyl, pyridylcarbonyl, C1-5alkylsulfonyl, phenylcarbonyl, phenylC1-5alkylcarbonyl, phenylsulfonyl, phenylC1-5alkylsulfonyl substituted phenylcarbonyl, substituted phenylC1-5alkylcarbonyl, substituted phenylsulfonyl, substituted phenylC1-5alkylsulfonyl, substituted phenyl, and substituted phenylC1-5alkyl [where the aromatic phenyl, phenylC1-5alkyl, phenylcarbonyl, phenylC1-5alkylcarbonyl, phenylsulfonyl, and phenylC1-5alkylsulfonyl substitutents are independently selected from one to five members of the group consisting of C1-5alkyl, C1-5alkoxy, hydroxy, halogen, trifluoromethyl, nitro, cyano, and amino]);
        
        
        W is selected from the group consisting of —CH═CH—, —S—, and —CH═N—;
        Q is selected from the group consisting of —CH═CH—, —S—, and —CH═N—;
        X is selected from the group consisting of carbonyl, C1-5alkyl, C1-5alkenyl, C1-5alkenylcarbonyl, and (CH2)m—C(O)— where m is 2–5;
        Y is selected from the group consisting of carbonyl, C1-5alkyl, C1-5alkenyl, C1-5alkenylcarbonyl, and (CH2)m—C(O)— where m is 2–5;
        n is 1, 2, or 3;
        Z is selected from the group consisting of hydroxy, C1-5 alkoxy, phenoxy, phenylC1-5alkoxy, amino, C1-5alkylamino, diC1-5alkylamino, phenylamino, phenylC1-5alkylamino, piperidin-1-yl
        
            substituted piperidin-1-yl (where the substituents are selected from the group consisting of C1-5alkyl, C1-5alkoxy, halo, aminocarbonyl, C1-5alkoxycarbonyl, and oxo;
            substituted phenylC1-5alkylamino (where the aromatic substitutents are selected from the group consisting of C1-5alkyl, C1-5alkoxy, phenylC1-5alkenyloxy, hydroxy, halogen, trifluoromethyl, nitro, cyano, and amino),
            substituted phenoxy (where the aromatic substitutents are selected from the group consisting of C1-5alkyl, C1-5alkoxy, hydroxy, halogen, trifluoromethyl, nitro, cyano, and amino),
            substituted phenylC1-5alkoxy (where the aromatic substitutents are selected from the group consisting of C1-5alkyl, C1-5alkoxy, hydroxy, halogen, trifluoromethyl, nitro, cyano, and amino),
            —OCH2CH2(OCH2CH2)sOCH2CH2O—, —NHCH2CH2(OCH2CH2)sOCH2CH2NH—, —NH(CH2)pO(CH2)qO(CH2)pNH—, —NH(CH2)qNCH3(CH2)sNH—, —NH(CH2)sNH—, and (NH(CH2)s)3N,
            
                where s, p, and q are independently selected from 1–7
            
            
            with the proviso that if n is 2, Z is not hydroxy, C1-5 alkoxy, amino, C1-5alkylamino, diC1-5alkylamino, phenylamino, phenylC1-5alkylamino, or piperidin-1-yl,
            
                with the further proviso that if n is 3, Z is (NH(CH2)s)3N.

and the salts thereof.

            
            
        
        
    
    




DETAILED DESCRIPTION OF THE INVENTION
The terms used in describing the invention are commonly used and known to those skilled in the art. “Independently” means that when there are more than one substituent, the substitutents may be different. The term “alkyl” refers to straight, cyclic and branched-chain alkyl groups and “alkoxy” refers O-alkyl where alkyl is as defined supra. “Cbz” refers to benzyloxycarbonyl. “Boc” refers to t-butoxycarbonyl and “Ts” refers to toluenesulfonyl. “DCC” refers to 1,3-dicyclohexylcarbodiimide, “DMAP” refers to 4-N′,N-dimethylaminopyridine and “HOBT” refers to 1-hydroxybenzotriazole hydrate. “Fmoc” refers to N-(9-fluorenylmethoxycarbonyl), “DABCO” refers to 1,4-Diazabicyclo[2.2.2]octane, “EDCI” refers to 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide, “Dde” refers to 1-(4,4-dimethyl-2,6-dioxocyclohexylidene)ethyl, and “TMOF” refers to trimethyl orthoformate. The side chains of α-amino acids refer to the substituents of the stereogenic carbon of an α-amino acid. For example if the amino acid is lysine, the side chain is 1-aminobutan-4-yl. The term natural amino acid refers to the 20 α-amino acids of the L configuration which are found in natural proteins. Unnatural α-amino acids include synthetic amino acids such as, α-aminoadipic acid, 4-aminobutanoic acid, 6-aminohexanoic acid, α-aminosuberic acid, 5-aminopentanoic acid, p-aminophenylalanine, α-aminopimelic acid γ-carboxyglutamic acid, p-carboxyphenylalanine, carnitine, citrulline, α,β-diaminopropionic acid, α,γ-diaminobutyric acid, homocitrulline, homoserine, and statine as well as D-configuration amino acids. The term “protectable group” refers to a hydroxy, amino, carboxy, carboxamide, guanidine, amidine or a thiol groups on an amino acid side. Compounds of the invention may be prepared by following general procedures known to those skilled in the art, and those set forth herein.
The compounds of the invention may be prepared by liquid phase organic synthesis techniques or by using amino acids which are bound to a number of known resins. The underlying chemistry, namely, acylation and alkylation reactions, peptide protection and deprotection reactions as well as peptide coupling reactions use similar conditions and reagents. The main distinction between the two methods is in the starting materials. While the starting materials for the liquid phase syntheses are the N-protected amino acids or the lower alkyl ester derivatives of either the N-protected or N-unprotected amino acids, the starting material for the resin syntheses are N-protected amino acids which are bound to resins by their carboxy termini.
General Procedure for the Solid-Phase Synthesis of Symmetrical Nα,Nα-Disubstituted Amino Acids
Scheme 1
An equivalent of an N-Fmoc-protected amino acid which is bound to a resin 1a is suspended in a suitable solvent such as DMF. This solvent is removed and the nitrogen protecting group (Fmoc) is removed by stirring the resin bound amino acid with an organic base, such as piperidine, and an addition portion of the solvent. A solution of about two to three equivalents of an appropriately substituted halide, 1b, and a suitable base such DIEA is added to the resin bound amino acid and this mixture is shaken for 18–36 h. The resulting mixture is washed with several portions of a suitable solvent and is suspended and shaken in an acidic solution, such as 50% TFA/CH2Cl2, over several hours to cleave the acid from the resin and give the N-disubstituted amino acid 1c.
By varying the resin bound amino acid 1a, one may obtain many of the compounds of the invention. The following resin bound amino acids may be used in Scheme I: alanine, N-g-(4-methoxy-2,3,6-trimethylbenzenesulfonyl)arginine, β-(4-methyltrityl)asparagine, aspartic acid (β-t-butyl ester), S-(trityl)cysteine, γ-(4-methyltrityl)glutamine, glutamic acid (β-t-butyl ester), glycine, N-imidazolyl-(trityl)histidine, isoleucine, leucine, N-ε-(2-chlorobenzyloxycarbonyl)lysine, N-ε-(t-butoxycarbonyl)lysine, methionine, phenylalanine, proline, O-(t-butyl)serine, O-(t-butyl)threonine, N-indolyl-(t-butoxycarbonyl)tryptophan, O-(t-butyl)tyrosine, valine, β-alanine, α-aminoadipic acid, 4-aminobutanoic acid, 6-aminohexanoic acid, α-aminosuberic acid, 5-aminopentanoic acid, p-aminophenylalanine, α-aminopimelic acid γ-carboxyglutamic acid, p-carboxyphenylalanine, carnitine, citrulline, α,β-diaminopropionic acid, α,γ-diaminobutyric acid, homocitrulline, homoserine, and statine. In addition, the choice of “W” and “X” can be varied by using known halide derivatives of 1b. For example using benzylchloride, 2-chloromethylthiophene, or 2-chloromethylpyridine gives compounds of the invention where “W” is —CH═CH—, —S—, or —CH═N—, respectively. For variations in “X”, the use of 2-chloroethylphenyl, 3-chloro-1-propenylbenzene, or benzeneacetyl chloride as 1b, give compounds where Y is (CH2)2, —CH═CH—CH2—, or —CH2C(O)— respectively. Still further, Scheme 1 may be used to produce combinatorial mixtures of products. Using mixtures of resin bound amino acids, 1a, with only one 1b produces said combinatorial mixtures. Alternatively, using one amino acid 1a with a mixture of 1b as well as mixture of 1a with mixtures of 1b gives a large range of combinatorial mixtures.




General Procedure for the Solid-Phase Synthesis of Unsymmetrical Nα,Nα-Disubstituted Amino Acids
Scheme 2, Step A
An equivalent of an N-Fmoc-protected amino acid which is bound to a resin 1a is suspended in a suitable solvent such as DMF. This solvent is removed and the nitrogen protecting group (Fmoc) is removed by stirring the resin bound amino acid with an organic base, such as piperidine, and an addition portion of the solvent. Trimethyl orthoformate and an appropriately substituted aldehyde 2a (5 equivalents) is added and the mixture is shaken under N2 overnight. This mixture is treated with a suspension of NaBH(OAc)3 (5 equivalents) in CH2Cl2 and shaken under N2 overnight. After filtration and washing with a suitable solvent, the resulting product, resin bound Nα-monosubstituted amino acid 2b, is rinsed with a suitable solvent and its identity is confirmed by MS and or HPLC analysis after treatment of a portion of the resin with 50% TFA/CH2Cl2.
Scheme 2, Step B
The resin 2b is suspended in an appropriate solvent such as DMF and is filtered. The appropriately substituted alkyl or arylkyl halide, 2c, and an appropriate base such as DIEA are added with some additional solvent and the mixture is shaken under N2 for 18–36 h. The resin bound Nα,Nα-disubstituted amino acid, 2d, is isolated from the suspension and the resin is cleaved with an acidic solution to give the free acid 2e.




Scheme 3, Step C
A resin bound amine, 2d, where R4 is nitro, is suspended in a suitable solvent, such as DMF, and is filtered. This mixture is treated with SnCl2 dihydrate in DMF and shaken under N2 overnight. The solvent is removed and the resin is washed successive portions of a suitable solvent to give the resin bound compound 3a where R4 is amino. The resin is suspended in a suitable solvent and is combined with an organic base, such as pyridine an appropriately substituted carboxylic acid anhydride, acid chloride, or sulfonyl chloride. The mixture is shaken under N2 overnight and is filtered to give the resin bound amino acid 3b. This material is treated with an acid and a suitable solvent to give the free amino acid 3b.
Scheme 3, Step D
The resin bound amine 3a is treated with TMOF and an appropriately substituted aldehyde 3c is added and the mixture is shaken under N2 overnight. The resulting mixture is drained and treated with a suspension of NaBH(OAc)3 in an appropriate solvent and this mixture is shaken under N2 overnight. The resin bound 3-aralkylaminophenyl amino acid is identified my spectral techniques after clevage to give the free acid 3d as previously described.
Scheme 3, Step E
Resin bound, 2d, where R1 is (CH2)4NH(Dde) is mixed with a suitable solvent, such as DMF, and shaken with successive portions of 2% solution of hydrazine hydrate in DMF over about 30 min. The resin is filtered and treated with a suitable solvent and a cyclic anhydride derivative 3e, and a base such as DMAP and pyridine. This mixture is shaken under N2 overnight and filtered to give the resin bound amine, 3f. This material is identified by spectral techniques after clevage to give the free acid 3f as previously described.




Scheme 4, Step F
Resin bound 2b, where R2 is nitro is suspended in CH2Cl2 and is treated with an organic base, such as pyridine, and 9-fluorenylmethoxy chloride. This mixture is shaken under N2 overnight, filtered and resuspended in a suitable solvent. This mixture is treated with SnCl2 dihydrate in DMF and shaken under N2 overnight. The solvent is removed and the resin is washed successive portions of a suitable solvent and filtered to give the resin bound compound 4a where R2 is amino. The resin 4a is then suspended in a suitable solvent, such as CH2Cl2, and is combined with 0.4 mmol of pyridine and 0.25–0.4 mmol of the appropriately substituted carboxylic acid anhydride, acid chloride, or sulfonyl chloride. The mixture is shaken under N2 overnight, filtered, and washed successively with three portions each of CH2Cl2 and MeOH. This resin is suspended in DMF, filtered, and shaken under N2 with 5 mL of a 40% solution of piperidine in DMF. After 1 h, the solvent is drained and the resin was washed successively with three portions each of suitable solvents to give the resin bound 4b. The identity of the compound was confirmed by spectral analysis after cleveage as previously described.




Scheme 5
The resin 2b (0.2 mmol) is suspended in CH2Cl2, filtered, and is resuspended in CH2Cl2. This suspension is treated with diethyl phosphonoacetic acid and diisopropylcarbodiimide or other suitable carbodiimide reagent, and the mixture is shaken under N2 overnight. The solvent is drained and the resulting resin 5a was washed successively with three portions each of CH2Cl2 and MeOH. The resin is suspended in DMF and filtered. A solution of the appropriately substituted aldehyde 5b (0.6–1.0 mmol) in 3–5 mL of DMF, lithium bromide (0.6–1.0 mmol), and a suitable base such as DIEA or Et3N (0.6–1.0 mmol) is added and the mixture is shaken under N2 overnight. The solvent is removed and the resin is washed successively with three portions each of DMF, CH2Cl2, and MeOH. The identity of the resin bound substituted amino acid 5c was confirmed spectral techniques. The resin bound material may be treated with 50% TFA/CH2Cl2 over 1–1.5 h, to give the acid 5c.




Scheme 6
To prepare compounds where n is 2 and Z is NH(CH2)sNH, products of Schemes 1–5 may be used in Scheme 6. Treatment of two equivalents of the substituted amino acid 1c with an equivalent of the diamine 6a, in the presence of HOBT and a peptide coupling agent such as EDCI and a base such as DIEA at room temperature over 16 h gives the dimer 6b.




General Procedure for the Solution-Phase Synthesis of Symmetrical Nα,Nα-Disubstituted Amino Acids
Scheme 7, Step A
A solution of of amino acid ester 7a, an appropriately substituted halide derivitive 1b, and an appropriate base such as DIEA, Na2CO3, or Cs2CO3 in a suitable solvent, such as DMF, is heated at 50–100° C. under N2 overnight, or until the starting material is exhausted, to give a mixture of the di and mono-substituted amines, 7b and 7c respectively. If the side chains of R1 contain acid cleavable protecting groups, those groups may be cleaved by treatment with 30–80% TFA/CH2Cl2. Esters 7b and 7c may be independently converted to the corresponding acids 7d and 7e by hydrolysis with an appropriate base such as aqueous NaOH.




General Procedure for the Solution-Phase Synthesis of Unsymmetrical Nα,Nα-Disubstituted Amino Acids
Scheme 8, Step A
A solution of 1 mmol of amino acid ester 8a (or the corresponding HCl salt and 1.1 mmol of DIEA) and 1–1.5 mmol of the appropriately substituted aldehyde 2a in 3–5 mL of trimethyl orthoformate was stirred at room temperature under N2 overnight. The solution was either concentrated and used directly for the next reaction, or was partitioned between EtOAc and water, washed with brine, dried over Na2SO4, and concentrated to give crude product, which was purified by MPLC to give mono-substituted product 8b.
Scheme 8, Step B
Amino ester 8b was dissolved in DMF, combined with 1.1–1.5 mmol of the appropriately substituted chloride or bromide 2c, and heated at 50–100° C. overnight. The reaction mixture was cooled and partitioned between water and EtOAc. The organic layer was washed three times with water and once with brine, dried over Na2SO4, and concentrated. The crude product was purified by MPLC to give pure 8c. For examples of 8c wherein the side chain R1 contained an acid-cleavable protecting group such as t-butylcarbamate, t-butyl ester, or t-butyl ether, 8c was stirred in 30–80% TFA/CH2Cl2 for 1–3 h. The reaction mixture was concentrated and optionally dissolved in HOAc and freeze-dried to give the deprotected form of 8c. For examples of 8c where R9 was equal to t-butyl, 8c was stirred in 30–80% TFA/CH2Cl2 for 1–3 h and treated as described above to give acid 8d. For examples of 8c where R9 was equal to methyl, ethyl, or other primary or secondary alkyl esters, 8c was stirred with with 1–2 mmol of aqueous LiOH, NaOH, or KOH in MeOH, EtOH, or THF at 20–80° C. until TLC indicated the absence of 8c. The solution was acidified to pH 4–5 with aqueous citric acid or HCl and was extracted with CH2Cl2 or EtOAc. The organic solution was washed with brine, dried over Na2SO4, and concentrated to give 8d.
Scheme 8, Step C
For examples of amino acid ester 8c where R1═(CH2)4NHBoc, 8c (1 mmol) was stirred in 30–80% TFA/CH2Cl2 for 1–3 h. The reaction mixture was concentrated to provide 8e as the TFA salt. Optionally, the TFA salt was dissolved in CH2Cl2 or EtOAc and washed with aqueous NaOH or Na2CO3, dried over Na2SO4, and concentrated to give 8e as the free base.
Scheme 8, Step D
A solution of 1 mmol of 8e, 1–4 mmol of an appropriate base such as DIEA, and 1–2 mmol of the appropriately substituted cyclic anhydride 3e was stirred in CH2Cl2 or DMF under N2 overnight. The resulting mixture was diluted with CH2Cl2 or EtOAc and washed with aqueous HCl, water, and brine, was dried over Na2SO4, and concentrated to provide 8f. Alternatively, 1 mmol of 8e, 1–4 mmol of an appropriate base such as DIEA, and 1–2 mmol of the appropriately substituted carboxylic acid anhydride (R11CO)2O or acid chloride R11COCl was stirred in CH2Cl2 or DMF under N2 overnight and worked up as above to provide 8g. Alternatively, 1 mmol of 8e, 1–4 mmol of an appropriate base such as DIEA, and 1–2 mmol of the appropriately substituted isocyanate R12NCO was stirred in CH2Cl2 or DMF under N2 overnight and worked up as above to provide 8h.




Scheme 9, Step A
For examples of 8c where R5═NO2, a solution of 1 mmol of 8c (where R2, R3, R4, or) and 10–12 mmol of SnCl2 dihydrate was stirred in MeOH, EtOH, or DMF at 20–80° C. for 0.5–24 h under N2. The solution was taken to room temperature and poured into aqueous Na2CO3 with rapid stirring. The resulting mixture was extracted with EtOAc or CH2Cl2 and the organic extracts were washed with brine, dried over Na2SO4, and concentrated to give the aminophenyl product 9a, which was purified by MPLC or used without further purification.
Scheme 9, Step B
A solution of 1 mmol of aminophenyl compound 9a and 1–1.5 mmol of the appropriately substituted aldehyde 2a in 3–5 mL of trimethyl orthoformate was stirred at room temperature under N2 overnight. The solution was either concentrated and used directly for the next reaction, or was partitioned between EtOAc and water, washed with brine, dried over Na2SO4, and concentrated to give crude product, which was purified by MPLC to give 9b. For examples of 9b wherein the side chain R1 or R9 contained an acid-cleavable protecting group such as t-butylcarbamate, t-butyl ester, or t-butyl ether, 9b was stirred in 30–80% TFA/CH2Cl2 for 1–3 h. The reaction mixture was concentrated and optionally dissolved in HOAc and freeze-dried to give the deprotected form of 9b.
Scheme 9, Step C
A solution of 1 mmol of 3-aminophenyl compound 9a, 1.1–2 mmol of pyridine, and 1–1.5 mmol of the appropriately substituted acid chloride, acid anhydride, or sulfonyl chloride in 3–5 mL of CH2Cl2 or ClCH2CH2Cl was stirred at room temperature under N2 overnight. The solution was partitioned between EtOAc and water, washed with water, saturated aqueous NaHCO3, and brine, dried over Na2SO4, and concentrated to give crude product which was optionally purified by MPLC to give amide or sulfonamide 9c. For examples of 9c wherein the side chain R1 or R9 contained an acid-cleavable protecting group such as t-butylcarbamate, t-butyl ester, or t-butyl ether, 9c was stirred in 30–80% TFA/CH2Cl2 for 1–3 h. The reaction mixture was concentrated and optionally dissolved in HOAc and freeze-dried to give the deprotected form of 9c.




General Procedure for the Solution-Phase Synthesis of Symmetrical Nα,Nα-Disubstituted Amino Amides and Their Dimers and Trimers
Scheme 10, Step A
A solution of 1 mmol of N-Cbz-protected amino acid 10a and the appropriate amine (ZH, 1 mmol), diamine (ZH2, 0.5 mmol), or triamine (ZH3 0.33 mmol), was treated with 1.1 mmol of HOBt, 1.1 mmol of DIEA, and 2.1 mmol of EDCI in 3–6 mL of CH2Cl2 or DMF. [Alternatively, 1 mmol of the pentafluorophenyl ester or N-hydroxysuccinimide ester of 10a was mixed with the appropriate portion of amine (ZH), diamine (ZH2), or triamine (ZH3) in 3–6 mL of DMF.] The solution was stirred at room temperature under N2 for 12–24 h, and EtOAc was added. The organic solution was washed with 5% aqueous citric acid, water, saturated NaHCO3, and brine, dried over Na2SO4, and concentrated. The crude product was optionally purified by MPLC to afford amide 10b. Compound 10b was stirred in 30–80% TFA/CH2Cl2 for 1–3 h. The reaction mixture was concentrated to provide the TFA salt which was dissolved in CH2Cl2 or EtOAc and washed with aqueous NaOH or Na2CO3, dried over Na2SO4, and concentrated to give 10c as the free base.
Scheme 10, Step B
A solution of 1 mmol of amino acid ester 10c (n=1), 2.5–3 mmol of the appropriately substituted chloride or bromide 2c, and 2.5–3 mmol of an appropriate base such as DIEA, Na2CO3, or Cs2CO3 in 3–5 mL of DMF was heated at 50–100° C. under N2 for 18–24 h. (For examples of 10c where n=2 or 3, the amounts of 2c and base were increased by two- or three-fold, respectively.) The reaction mixture was cooled and partitioned between water and EtOAc. The organic layer was washed three times with water and once with brine, dried over Na2SO4, and concentrated. The crude product was purified by MPLC to give pure amide 10d.
Alternatively, a solution of 1 mmol of amino acid ester 10c (n=1), 2.5–3 mmol of the appropriately substituted aldehyde 2a, and 2.5–3 mmol of borane-pyridine complex in 3–5 mL of DMF or EtOH was stirred at room temperature under N2 for 3–5 days. (For examples of 10c where n=2 or 3, the amounts of 2c and borane-pyridine complex were increased by two- or three-fold, respectively.) The mixture was concentrated to dryness and was partitioned between water and CH2Cl2, washed with brine, dried over Na2SO4, and concentrated. The crude product was purified by MPLC to give pure amide 10d.
Scheme 10, Step C
For examples of 10d where R1═CH2CH2CO2-t-Bu or CH2CO2-t-Bu, 10d was stirred in 30–80% TFA/CH2Cl2 for 1–24 h. The reaction mixture was concentrated and optionally dissolved in HOAc and freeze-dried to give acid 10e.
Scheme 10, Step D
For examples of 10d where R1 is equal to (CH2)4NHBoc, 10d was stirred in 30–80% TFA/CH2Cl2 for 1–24 h. The reaction mixture was concentrated and optionally dissolved in HOAc and freeze-dried to give amine 10f as the TFA salt which was optionally dissolved in CH2Cl2 or EtOAc, washed with aqueous NaOH or Na2CO3, dried over Na2SO4, and concentrated to give 10f as the free base.
Scheme 10, Step E
A solution of 1 mmol of 10f, 1–4 mmol of an appropriate base such as DIEA, and 1–2 mmol of the appropriately substituted cyclic anhydride 3e was stirred in CH2Cl2 or DMF under N2 overnight. The resulting mixture was diluted with CH2Cl2 or EtOAc and washed with aqueous HCl, water, and brine, was dried over Na2SO4, and concentrated to provide acid 10g. Alternatively, 1 mmol of 10f, 1–4 mmol of an appropriate base such as DIEA, and 1–2 mmol of the appropriately substituted carboxylic acid anhydride (R11CO)2O or acid chloride R11COCl was stirred in CH2Cl2 or DMF under N2 overnight and worked up as above to provide 10h. Alternatively, 1 mmol of 8e, 1–4 mmol of an appropriate base such as DIEA, and 1–2 mmol of the appropriately substituted isocyanate R12NCO was stirred in CH2Cl2 or DMF under N2 overnight and worked up as above to provide 10i.




General Procedure for the Solid-Phase Synthesis of Nα,Nα-Bis-Cinnamyl Amino Acids and Nα-Cinnamyl Amino Acids
Scheme 11
An equivalent of an N-Fmoc-protected amino acid 11a which is bound to a polystyrene resin such as Wang resin is suspended in a suitable solvent such as DMF. This solvent is removed and the nitrogen protecting group (Fmoc) is removed by stirring the resin bound amino acid with an organic base, such as piperidine, and an addition portion of the solvent. After filtration and washing with solvent, the resin is suspended in an appropriate solvent such as DMF. A solution of about 2–3 equivalents of an appropriately substituted halide 11b and a suitable base such DIEA is added to the resin bound amino acid and this mixture is shaken for 18–36 h. The resulting mixture is washed with several portions of a suitable solvent and is suspended and shaken in an acidic solution, such as 50% TFA/CH2Cl2, over several hours to cleave the acid from the resin to give a mixture of the Nα,Nα-bis-cinnamyl amino acid 11c and the Nα-cinnamyl amino acid 11d.
By varying the resin bound amino acid 11a, one may obtain many of the compounds of the invention. The following resin bound amino acids may be used in Scheme 11: alanine, N-g-(4-methoxy-2,3,6-trimethylbenzenesulfonyl)arginine, β-(4-methyltrityl)asparagine, aspartic acid (β-t-butyl ester), S-(trityl)cysteine, γ-(4-methyltrityl)glutamine, glutamic acid (β-t-butyl ester), glycine, N-imidazolyl-(trityl)histidine, isoleucine, leucine, N-ε-(2-chlorobenzyloxycarbonyl)lysine, N-ε-(t-butoxycarbonyl)lysine, methionine, phenylalanine, proline, O-(t-butyl)serine, O-(t-butyl)threonine, N-indolyl-(t-butoxycarbonyl)tryptophan, O-(t-butyl)tyrosine, valine, β-alanine, α-aminoadipic acid, 4-aminobutanoic acid, 6-aminohexanoic acid, α-aminosuberic acid, 5-aminopentanoic acid, p-aminophenylalanine, α-aminopimelic acid γ-carboxyglutamic acid, p-carboxyphenylalanine, carnitine, citrulline, α,β-diaminopropionic acid, α,γ-diaminobutyric acid, homocitrulline, homoserine, and statine.




Scheme 12, Step A
An equivalent of an N-Fmoc-protected amino acid which is bound to a resin 11a is suspended in a suitable solvent such as DMF. This solvent is removed and the nitrogen protecting group (Fmoc) is removed by stirring the resin bound amino acid with an organic base, such as piperidine, and an addition portion of the solvent. After filtration and washing with solvent, the resin is suspended in an appropriate solvent such as trimethyl orthoformate (TMOF), an appropriately substituted aldehyde 12a (5 equivalents) is added, and the mixture is shaken under N2 overnight. This mixture is treated with a suspension of NaBH(OAc)3 (5 equivalents) in CH2Cl2 and shaken under N2 overnight. After filtration and washing with a suitable solvent, the resulting product, resin bound Nα-monosubstituted amino acid 12b, is suspended and shaken in an acidic solution, such as 50% TFA/CH2Cl2, over several hours to cleave the acid from the resin to give the Nα-cinnamyl amino acid 11d.
Scheme 12, Step B
The resin 12b is suspended in an appropriate solvent such as DMF and is filtered. The appropriately substituted halide 12c and an appropriate base such as DIEA are added with some additional solvent and the mixture is shaken under N2 for 18–36 h. The resin bound Nα,Nα-cinnamyl amino acid 12d is isolated from the suspension and the resin is cleaved with an acidic solution as described above to give the free acid 12e.




General Procedure for the Solution-Phase Synthesis of Nα,Nα-Bis-Cinnamyl Amino Acids and Nα-Cinnamyl Amino Acids
Scheme 13
A solution of of amino acid ester 13a, an appropriately substituted halide 11b, and an appropriate base such as DIEA, Na2CO3, or Cs2CO3 in a suitable solvent, such as DMF, is heated at 50–100° C. under N2 overnight, or until the starting material is exhausted, to give a mixture of the Nα,Nα-bis-cinnamyl amino acid ester 13b and Nα-cinnamyl amino acid ester 13c. If the side chain of R1 contains an acid-cleavable protecting group such as t-butylcarbamate, t-butyl ester, or t-butyl ether, those groups may be cleaved by treatment with an acidic solution such as 30–80% TFA/CH2Cl2 or 2–4N HCl in EtOAc. For examples of 13b and 13c where the ester group R4 is a primary alkyl group such as methyl or ethyl, esters 13b and 13c may be independently converted to the corresponding acids 11c and 11d by hydrolysis with an appropriate base such as aqueous NaOH, KOH, or LiOH. For examples of 13b and 13c where the ester group R4 is an acid-cleavable group such as t-butyl, esters 13b and 13c may be independently converted to the corresponding acids 11c and 11d by treatment with an acidic solution such as 30–80% TFA/CH2Cl2 or 2–4N HCl in EtOAc.




Scheme 14, Step A
A solution of 1 mmol of amino acid ester and 1–1.5 mmol of the appropriately substituted aldehyde 12a in 3–5 mL of TMOF was stirred at room temperature under N2 overnight. The solution was concentrated and used directly for the next reaction; optionally, the solution was partitioned between EtOAc and water, washed with brine, dried over Na2SO4, and concentrated to give crude product, which was purified by MPLC to give mono-substituted product 14a. For examples of 14a wherein the side chain R1 contained an acid-cleavable protecting group such as t-butylcarbamate, t-butyl ester, or t-butyl ether, 8c was treated with an acidic solution such as 30–80% TFA/CH2Cl2 or 2–4N HCl in EtOAc. The reaction mixture was concentrated and optionally dissolved in HOAc and freeze-dried to give the deprotected form of 14a. For examples of 14a where the ester group R4 is a primary alkyl group such as methyl or ethyl, esters 14a may be converted to the corresponding acids 11d by hydrolysis with an appropriate base such as aqueous NaOH, KOH, or LiOH. For examples of 14a where the ester group R4 is an acid-cleavable group such as t-butyl, esters 14a may be converted to the corresponding acids 11d by treatment with an acidic solution such as 30–80% TFA/CH2Cl2 or 2–4N HCl in EtOAc.
Scheme 14, Step B
Amino ester 14a was dissolved in DMF, combined with 1.1–1.5 mmol of the appropriately substituted chloride or bromide 12c, and heated at 50–100° C. overnight. The reaction mixture was cooled and partitioned between water and EtOAc. The organic layer was washed with water and brine, dried over Na2SO4, and concentrated. The crude product was purified by MPLC to give pure 14b. For examples of 14b wherein the side chain R1 contained an acid-cleavable protecting group such as t-butylcarbamate, t-butyl ester, or t-butyl ether, 8c was treated with an acidic solution such as 30–80% TFA/CH2Cl2 or 2–4N HCl in EtOAc. The reaction mixture was concentrated and optionally dissolved in HOAc and freeze-dried to give the deprotected form of 14b. For examples of 14b where the ester group R4 is a primary alkyl group such as methyl or ethyl, esters 14b may be converted to the corresponding acids 12e by hydrolysis with an appropriate base such as aqueous NaOH, KOH, or LiOH. For examples of 14b where the ester group R4 is an acid-cleavable group such as t-butyl, esters 14b may be converted to the corresponding acids 12e by treatment with an acidic solution such as 30–80% TFA/CH2Cl2 or 2–4N HCl in EtOAc.




Although the claimed compounds are useful as competitive binders to the EPO receptor, some compounds are more active than others and are either preferred or particularly preferred.







The particularly preferred “R1”s are the side chain of lysine, ornithine, arginine, aspartic acid, glutamic acid, glutamine, cysteine, methionine, serine, and threonine.
The particularly preferred “R2 and R3”s are phenoxy, substituted phenoxy, benzyloxy, and substituted benzyloxy.
The particularly preferred “R4 and R5”s are phenoxy, substituted phenoxy, benzyloxy, and substituted benzyloxy.
The particularly preferred “W” is —CH═CH—
The particularly preferred “Q” is —CH═CH—
The particularly preferred “X” are C1-5alkenyl and CH2.
The particularly preferred “Y” are C1-5alkenyl and CH2.
The particularly preferred “n” are 1 and 2.
The particularly preferred “Z” are hydroxy, methoxy, phenethylamino, substituted phenethylamino, and —NH(CH2)2O(CH2)2O(CH2)2NH—.
Pharmaceutically useful compositions the compounds of the present invention, may be formulated according to known methods such as by the admixture of a pharmaceutically acceptable carrier. Examples of such carriers and methods of formulation may be found in Remington's Pharmaceutical Sciences. To form a pharmaceutically acceptable composition suitable for effective administration, such compositions will contain an effective amount of the compound of the present invention.
Therapeutic or diagnostic compositions of the invention are administered to an individual in amounts sufficient to treat or diagnose disorders in which modulation of EPO receptor-related activity is indicated. The effective amount may vary according to a variety of factors such as the individual's condition, weight, sex and age. Other factors include the mode of administration. The pharmaceutical compositions may be provided to the individual by a variety of routes such as subcutaneous, topical, transdermal, oral and parenteral.
The term “chemical derivative” describes a molecule that contains additional chemical moieties which are not normally a part of the base molecule. Such moieties may improve the solubility, half-life, absorption, etc. of the base molecule. Alternatively the moieties may attenuate undesirable side effects of the base molecule or decrease the toxicity of the base molecule. Examples of such moieties are described in a variety of texts, such as Remington's Pharmaceutical Sciences.
Compounds disclosed herein may be used alone at appropriate dosages defined by routine testing in order to obtain optimal inhibition of the EPO receptor or its activity while minimizing any potential toxicity. In addition, co-administration or sequential administration of other agents may be desirable.
The present invention also has the objective of providing suitable topical, transdermal, oral, systemic and parenteral pharmaceutical formulations for use in the novel methods of treatment of the present invention. The compositions containing compounds according to this invention as the active ingredient for use in the modulation of EPO receptors can be administered in a wide variety of therapeutic dosage forms in conventional vehicles for administration. For example, the compounds or modulators can be administered in such oral dosage forms as tablets, capsules (each including timed release and sustained release formulations), pills, powders, granules, elixirs, tinctures, solutions, suspensions, syrups and emulsions, or by transdermal delivery or injection. Likewise, they may also be administered in intravenous (both bolus and infusion), intraperitoneal, subcutaneous, topical with or without occlusion, transdermal, or intramuscular form, all using forms well known to those of ordinary skill in the pharmaceutical arts. The compounds of the present invention may be delivered by a wide variety of mechanisms, including but not limited to, transdermal delivery, or injection by needle or needle-less injection means. An effective but non-toxic amount of the compound desired can be employed as an EPO receptor modulating agent.
The daily dosage of the products may be varied over a wide range from 0.01 to 1,000 mg per patient, per day. For oral administration, the compositions are preferably provided in the form of scored or unscored tablets containing 0.01, 0.05, 0.1, 0.5, 1.0, 2.5, 5.0, 10.0, 15.0, 25.0, and 50.0 milligrams of the active ingredient for the symptomatic adjustment of the dosage to the patient to be treated. An effective amount of the drug is ordinarily supplied at a dosage level of from about 0.0001 mg/kg to about 100 mg/kg of body weight per day. The range is more particularly from about 0.001 mg/kg to 10 mg/kg of body weight per day. The dosages of the EPO receptor modulators are adjusted when combined to achieve desired effects. On the other hand, dosages of these various agents may be independently optimized and combined to achieve a synergistic result wherein the pathology is reduced more than it would be if either agent were used alone.
Advantageously, compounds or modulators of the present invention may be administered in a single daily dose, or the total daily dosage may be administered in divided doses of two, three or four times daily. Furthermore, compounds or modulators for the present invention can be administered in intranasal form via topical use of suitable intranasal vehicles, or via transdermal routes, using those forms of transdermal skin patches well known to those of ordinary skill in that art. To be administered in the form of a transdermal delivery system, the dosage administration will, of course, be continuous rather than intermittent throughout the dosage regimen.
For combination treatment with more than one active agent, where the active agents are in separate dosage formulations, the active agents can be administered concurrently, or they each can be administered at separately staggered times.
The dosage regimen utilizing the compounds or modulators of the present invention is selected in accordance with a variety of factors including type, species, age, weight, sex and medical condition of the patient; the severity of the condition to be treated; the route of administration; the renal and hepatic function of the patient; and the particular compound thereof employed. A physician or veterinarian of ordinary skill can readily determine and prescribe the effective amount of the drug required to prevent, counter or arrest the progress of the condition. Optimal precision in achieving concentrations of drug within the range that yields efficacy without toxicity requires a regimen based on the kinetics of the drug's availability to target sites. This involves a consideration of the distribution, equilibrium, and elimination of a drug.
In the methods of the present invention, the compounds or modulators herein described in detail can form the active ingredient, and are typically administered in admixture with suitable pharmaceutical diluents, excipients or carriers (collectively referred to herein as “carrier” materials) suitably selected with respect to the intended form of administration, that is, oral tablets, capsules, elixirs, syrups and the like, and consistent with conventional pharmaceutical practices.
For instance, for oral administration in the form of a tablet or capsule, the active drug component can be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water and the like. Moreover, when desired or necessary, suitable binders, lubricants, disintegrating agents and coloring agents can also be incorporated into the mixture. Suitable binders include, without limitation, starch, gelatin, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tragacanth or sodium alginate, carboxymethylcellulose, polyethylene glycol, waxes and the like. Lubricants used in these dosage forms include, without limitation, sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride and the like. Disintegrators include, without limitation, starch, methyl cellulose, agar, bentonite, xanthan gum and the like.
For liquid forms the active drug component can be combined in suitably flavored suspending or dispersing agents such as the synthetic and natural gums, for example, tragacanth, acacia, methyl-cellulose and the like. Other dispersing agents which may be employed include glycerin and the like. For parenteral administration, sterile suspensions and solutions are desired. Isotonic preparations which generally contain suitable preservatives are employed when intravenous administration is desired.
Topical preparations containing the active drug component can be admixed with a variety of carrier materials well known in the art, such as, e.g., alcohols, aloe vera gel, allantoin, glycerine, vitamin A and E oils, mineral oil, PPG2 myristyl propionate, and the like, to form, e.g., alcoholic solutions, topical cleansers, cleansing creams, skin gels, skin lotions, and shampoos in cream or gel formulations.
The compounds or modulators of the present invention can also be administered in the form of liposome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles and multilamellar vesicles. Liposomes can be formed from a variety of phospholipids, such as cholesterol, stearylamine or phosphatidylcholines.
Compounds of the present invention may also be delivered by the use of monoclonal antibodies as individual carriers to which the compound molecules are coupled. The compounds or modulators of the present invention may also be coupled with soluble polymers as targetable drug carriers. Such polymers can include polyvinyl-pyrrolidone, pyran copolymer, polyhydroxypropylmethacryl-amidephenol, polyhydroxy-ethylaspartamidephenol, or polyethyl-eneoxidepolylysine substituted with palmitoyl residues. Furthermore, the compounds or modulators of the present invention may be coupled to a class of biodegradable polymers useful in achieving controlled release of a drug, for example, polylactic acid, polyepsilon caprolactone, polyhydroxy butyric acid, polyorthoesters, polyacetals, polydihydro-pyrans, polycyanoacrylates and cross-linked or amphipathic block copolymers of hydrogels, and other suitable polymers known to those skilled in the art.
For oral administration, the compounds or modulators may be administered in capsule, tablet, or bolus form or alternatively they can be mixed in the animals feed. The capsules, tablets, and boluses are comprised of the active ingredient in combination with an appropriate carrier vehicle such as starch, talc, magnesium stearate, or di-calcium phosphate. These unit dosage forms are prepared by intimately mixing the active ingredient with suitable finely-powdered inert ingredients including diluents, fillers, disintegrating agents, and/or binders such that a uniform mixture is obtained. An inert ingredient is one that will not react with the compounds or modulators and which is non-toxic to the animal being treated. Suitable inert ingredients include starch, lactose, talc, magnesium stearate, vegetable gums and oils, and the like. These formulations may contain a widely variable amount of the active and inactive ingredients depending on numerous factors such as the size and type of the animal species to be treated and the type and severity of the infection. The active ingredient may also be administered as an additive to the feed by simply mixing the compound with the feedstuff or by applying the compound to the surface of the feed. Alternatively the active ingredient may be mixed with an inert carrier and the resulting composition may then either be mixed with the feed or fed directly to the animal. Suitable inert carriers include corn meal, citrus meal, fermentation residues, soya grits, dried grains and the like. The active ingredients are intimately mixed with these inert carriers by grinding, stirring, milling, or tumbling such that the final composition contains from 0.001 to 5% by weight of the active ingredient.
The compounds or modulators may alternatively be administered parenterally via injection of a formulation consisting of the active ingredient dissolved in an inert liquid carrier. Injection may be either intramuscular, intraruminal, intratracheal, or subcutaneous, either by needle or needle-less means. The injectable formulation consists of the active ingredient mixed with an appropriate inert liquid carrier. Acceptable liquid carriers include the vegetable oils such as peanut oil, cotton seed oil, sesame oil and the like as well as organic solvents is such as solketal, glycerol formal and the like. As an alternative, aqueous parenteral formulations may also be used. The vegetable oils are the preferred liquid carriers. The formulations are prepared by dissolving or suspending the active ingredient in the liquid carrier such that the final formulation contains from 0.005 to 10% by weight of the active ingredient.
Topical application of the compounds or modulators is possible through the use of a liquid drench or a shampoo containing the instant compounds or modulators as an aqueous solution or suspension. These formulations generally contain a suspending agent such as bentonite and normally will also contain an antifoaming agent. Formulations containing from 0.005 to 10% by weight of the active ingredient are acceptable. Preferred formulations are those containing from 0.01 to 5% by weight of the instant compounds or modulators.
The compounds of Formula I may be used in pharmaceutical compositions to treat patients (humans and other mammals) with disorders or conditions associated with the production of erythropoietin or modulated by the EPO receptor. The compounds can be administered in the manner of the commercially available product or by any oral or parenteral route (including but not limited to, intravenous, intraperitoneal, intramuscular, subcutaneous, dermal patch), where the preferred route is by injection. When the method of administration is intravenous infusion, compound of Formula I may be administered in a dose range of about 0.01 to 1 mg/kg/min. For oral administration, the dose range is about 0.1 to 100 mg/kg.
The pharmaceutical compositions can be prepared using conventional pharmaceutical excipients and compounding techniques. Oral dosage forms may be used and are elixirs, syrups, capsules, tablets and the like. Where the typical solid carrier is an inert substance such as lactose, starch, glucose, methyl cellulose, magnesium stearate, dicalcium phosphate, mannitol and the like; and typical liquid oral excipients include ethanol, glycerol, water and the like. All excipients may be mixed as needed with disintegrants, diluents, granulating agents, lubricants, binders and the like using conventional techniques known to those skilled in the art of preparing dosage forms. Parenteral dosage forms may be prepared using water or another sterile carrier.
Typically the compounds of Formula I are isolated as the free base, however when possible pharmaceutically acceptable salts can be prepared. Examples of such salts include hydrobromic, hydroiodic, hydrochloric, perchloric, sulfuric, maleic, fumaric, malic, tartaric, citric, benzoic, mandelic, methanesulfonic, hydroethanesulfonic, benzenesulfonic, oxalic, pamoic, 2-naphthalenesulfonic, p-toluenesulfonic, cyclohexanesulfamic and saccharic.
In order to illustrate the invention the following examples are included. These examples do not limit the invention. They are only meant to suggest a method of practicing the invention. Those knowledgeable in chemical synthesis and the treatment of EPO related disorders may find other methods of practicing the invention. However those methods are deemed to be within the scope of this invention.
BIOLOGICAL EXAMPLES
The compounds of the invention were evaluated for the ability to compete with EPO in the following immobilized EPO receptor preparation (EBP-Ig, EPO binding protein-Ig).
EBP-Ig fusion protein (as disclosed in WO97/27219 which is herein incorporated by reference) was purified by affinity chromatography from the conditioned media of NSO cells engineered to express a recombinant gene construct which functionally joined the N-terminal 225 amino acids of the human EPO receptor and an Ig heavy chain as described herein. The interaction of biotin and streptavidin is frequently employed to capture and effectively immobilize reagents useful in assay protocols and has been employed here as a simple method to capture and immobilize EBP-Ig. EBP-Ig is initially randomly modified with an amine reactive derivative of biotin to produce biotinylated-EBP-Ig. Use of streptavidin coated plates allows the capture of the biotinylated EBP-Ig on the surface of a scintillant impregnated coated well (Flash plates, NEN-DuPont). Upon binding of [125I]EPO to the ligand binding domain, specific distance requirements are satisfied and the scintillant is induced to emit light in response to the energy emitted by the radioligand. Unbound radioligand does not produce a measurable signal because the energy from the radioactive decay is too distant from the scintillant. The amount of light produced was quantified to estimate the amount of ligand binding. The specific assay format was suitable for the multi-well plate capacity of a Packard TopCount Microplate Scintillation counter. Compounds which were capable of reducing the amount of detected signal through competitive binding with the radioligand were identified.
Biotinylated EBP-Ig was prepared as follows. EBP-Ig (3 mL, OD280 2.9) was exchanged into 50 mM sodium bicarbonate, pH 8.5 using a Centriprep 10 ultrafiltration device. The final volume of the exchanged protein was 1.2 mL (OD280 2.6, representing about 2 mg total protein). 10 μL of a 4 mg/ml solution of NHS-LC-Biotin (Pierce) was added and the reaction mixture placed on ice in the dark for two hours. Unreacted biotin was removed by exchange of the reaction buffer into PBS in a Centriprep 10 device and the protein reagent aliquoted and stored at −70° C.
Each individual binding well (200 μL) contained final concentrations of 1 μg/mL of biotinylated EBP-Ig, 0.5 nM of [125I]EPO (NEN Research Products, Boston, 100 μCi/μg) and 0–500 μM of test compound (from a 10–50 mM stock in 100% DMSO). All wells were adjusted to a final DMSO concentration of 5%. All assay points were performed in triplicate and with each experiment a standard curve for unlabelled EPO was performed at final concentration of 2000, 62, 15, 8, 4, and 0 nM. After all additions were made, the plate was covered with an adhesive top seal and placed in the dark at room temperature overnight. The next day all liquid was aspirated from the wells to limit analyte dependent quench of the signal, and the plates were counted on a Packard TOPCOUNT Microplate Scintillation Counter. Non-specific binding (NSB) was calculated as the mean CPM of the 2000 nM EPO wells and total binding (TB) as the mean of the wells with no added unlabelled EPO. Corrected total binding (CTB) was calculated as: TB−NSB=CTB. The concentration of test compound which reduced CTB to 50% was reported as the IC50. Typically the IC50 value for unlabelled EPO was ca. 2–7 nM and EMP1 was 0.1 μM. Table 1 lists the average % inhibition, and if determined the IC50 and IC30 values for compounds of Formula I, where the compound numbers refer to the compounds in the tables accompanying the preparative examples.







TABLE 1







Inhibition of EPO binding to EBP-Ig












cpd
% inh @ 50 μM
IC30 μM*
IC50, μM*
















11
70
nd
nd



12
59
nd
nd



14
30
nd
nd



15
48
nd
nd



77
52
30
40



82
32
nd
nd



86
37
nd
nd



100
34
nd
nd



101
32
nd
nd



104
78
10
30



105
70
25
35



107
78
30
42



108
81
23
36



110
54
6
10



112
59
2
10



114
37
10
nd



115
35
nd
nd



116
32
nd
nd



117
34
nd
nd



118
36
2
10



119
34
nd
nd



120
35
nd
nd



121
45
6
nd



137
60
5
30



139
46
2
10



178
36
nd
nd



179
30
nd
nd



183
36
nd
nd



184
53
10
nd



203
37
50
nd



211
62
20
65



220
45
30
50



221
48
10
80



222
56
5
nd



224
51
25
50



227
48
20
50



230
42
nd
nd



231
36
nd
nd



235
49
20
50



237
55
30
70



238
39
nd
nd



239
46
8
50



243
75
2
18



244
66
1
28



246
79
10
75



247
47
7
18



248
56
7
20



249
72
7
10



250
78
7
20



251
49
10
45



261
51
1.5
2



262
93
1
1.5



263
88
1
1.5



264
89
1.5
8



265
65
1
6



266
82
1
4



267
83
2
6



268
40
nd
nd



269
55
8
85



270
56
7
100



271
77
2
7



272
78
5
10



285
41
nd
nd



286
46
35
65



287
36
nd
nd



300
57
35
145



305
48
35
225



312
45
10
85



321
42
45
nd



363
33
35
220



366
38
65
nd



368
40
90
nd







*nd = not determined






PREPARATIVE EXAMPLES
Unless otherwise noted, materials used in the examples were obtained from commercial suppliers and were used without further purification. Melting points were determined on a Thomas Hoover apparatus and are uncorrected. Proton nuclear magnetic resonance (1H NMR) spectra were measured in the indicated solvent with tetramethylsilane (TMS) as the internal standard using a Bruker AC-300 NMR spectrometer. NMR chemical shifts are expressed in parts per million (ppm) downfield from internal TMS using the d scale. 1H NMR data are tabulated in order: multiplicity, (s, singlet; d, doublet; t, triplet; q, quartet; m, multiplet), number of protons, coupling constant in Hertz). Electrospray (ES) mass spectra (MS) were determined on a Hewlett Packard Series 1090 LCMS Engine. Elemental analyses were performed by Quantitative Technologies, Inc. (QTI), PO Box 470, Salem Industrial Park, Bldg #5, Whitehouse, N.J. 08888-0470. Analytical thin layer chromatography (TLC) was done with Merck Silica Gel 60 F254 plates (250 micron). Medium pressure liquid chromatography (MPLC) was done with Merck Silica Gel 60 (230–400 mesh).
Example 1
N,N-bis(3-Phenoxycinnamyl)Glu(O-t-Bu)-OMe (cpd 96) and N-(3-phenoxycinnamyl)Glu(O-t-Bu)-OMe (cpd 334)
A solution of 500 mg (1.97 mmol) of H-Glu(O-t-Bu)OMe.HCl, 997 mg (3.45 mmol) of 3-phenoxycinnamyl bromide (Jackson, W. P.; Islip, P. J.; Kneen, G.; Pugh, A.; Wates, P. J. J. Med. Chem. 31 1988; 499–500), and 0.89 mL (5.1 mmol, 660 mg) of DIEA in 5 mL of DMF was stirred under N2 at room temperature for 40 h. The mixture was partitioned between EtOAc and water and the organic layer was washed with water and brine. After drying over Na2SO4, the organic solution was concentrated to give 1.24 g of orange oil. The crude residue was purified by MPLC using a solvent gradient ranging from 10–30% EtOAc/hexanes to give two products. The less polar product (cpd 96, 235 mg, 19% based on starting amino acid), was isolated as a pale yellow oil; 1H NMR (CDCl3, 300 MHz) 1.39 (s, 9H), 2.0 (m, 2H), 2.33 (dt, 2H, J=2, 7 Hz), 3.24 (dd, 2H, J=8, 15 Hz), 3.5, (m, 3H), 3.69 (s, 3H), 6.13 (m, 2H), 6.47 (d, 2H, J=16 Hz), 6.86 (dd, 2H, J=1.5, 8 Hz), 7.0–7.4 (complex, 16H); MS (ES+) m/z 634 (MH+).
The more polar product (cpd 334, 422 mg, 50% based on starting amino acid) was isolated as a pale yellow oil; 1H NMR (CDCl3, 300 MHz) 1.42 (s, 9H), 1.9 (m, 2H), 2.35 (t, 2H, J=7.5 Hz), 3.2–3.4 (complex, 3H), 3.71 (s, 3H), 6.17 (dt, 1H, J=16, 6 Hz), 6.46 (d, 1H, J=16 Hz), 6.87 (dd, 1H, J=1.5, 8 Hz), 7.01 (m, 3H), 7.10 (t, 2H, J=7.5 Hz), 7.2–7.4 (complex, 3H); MS (ES+) m/z 426 (MH+). Anal. Calcd for C25H31NO5: C, 70.57; H, 7.34; N, 3.29. Found: C, 70.29; H, 7.14; N, 3.08.
Example 2
N-(3-Phenoxycinnamyl)Glu-OMe (cpd 325)
A solution of 95 mg (0.22 mmol) of N-(3-phenoxycinnamyl)Glu(O-t-Bu)-OMe (cpd 334) in 3 mL of 50% TFA/CH2Cl2 was stirred for 2 h at room temperature. The mixture was concentrated and the residue was dissolve in acetic acid and freeze-dried to give 117 mg of N-(3-phenoxycinnamyl)Glu-OMe (cpd 325)as an off-white solid; 1H NMR (CD3OD, 300 MHz) 2.3–2.7 (complex, 4H), 3.78 (s, 3H), 3.81 (d, 2H, J=7 Hz), 4.09 (t, 1H, J=5 Hz), 6.17 (dt, 1H, J=16, 7 Hz), 6.55 (d, 1H, J=16 Hz), 6.9 (m, 4H), 7.11 (t, 2H, J=7.5 Hz), 7.3 (m, 4H); MS (ES+) m/z 370 (MH+), 209. Anal. Calcd for C21H23NO5.C2HF3O2: C, 57.14; H, 5.00; N, 2.90. Found: C, 57.07; H, 5.02; N, 2.73.
Example 3
N,N-bis(3-Phenoxycinnamyl)Asp(O-t-Bu)-O-t-Bu (cpd 106)
A solution of 1.00 g (3.55 mmol) of Asp(O-t-Bu)-O-t-Bu.HCl, 2.05 g (7.1 mmol) of 3-phenoxycinnamyl bromide, and 1.86 mL (10.7 mmol, 1.38 g) of DIEA in 15 mL of DMF was heated under N2 at 60° C. overnight. Additional 3-phenoxycinnamyl bromide (1.0 g, 3.4 mmol) and DIEA (0.95 mL, 0.705 g, 5.4 mmol) were added and heating was continued for an additional 14 h. The mixture was cooled and partitioned between EtOAc and water. The organic layer was washed twice with water, once with brine, and was dried over Na2SO4. The solution was concentrated to give 3.5 g of an amber oil which was purified by MPLC using a solvent gradient ranging from 2.5–3% EtOAc/hexanes to afford 1.21 g of cpd 106 as a pale yellow oil; 1H NMR (CDCl3, 300 MHz) 1.41 (s, 9H), 1.48 (s, 9H), 2.49 (dd, 1H, J=7.5, 15.5 Hz), 2.70 (dd, 1H, J=7.5, 15.5 Hz), 3.26 (dd, 2H, J=7.5, 14.5 Hz), 3.47 (dd, 2H, J=4, 14.5 Hz), 3.88 (t, 1H, J=7.5), 6.13 (m, 2H), 6.48 (d, 2H, J=16 Hz), 6.86 (dd, 2H, J=2, 8 Hz), 7.0 (m, 6H), 7.1 (m, 4H), 7.2–7.4 (complex, 6H); MS (ES+) m/z 662 (MH+).
Example 4
N,N-bis(3-Phenoxycinnamyl)Asp-OH (cpd 107)
A solution of 1.14 g (1.62 mmol) of cpd 106 in 16 mL of 50% TFA/CH2Cl2 was stirred at room temperature for 24 h. The solution was concentrated and pumped to give 1.37 g ( ̃100%) cpd 107 as an amber oil; 1H NMR (CD3OD, 300 MHz) 3.1 (m, 2H), 4.0 (dd, 2H, J=8, 14 Hz), 4.27 (dd, 2H, J=8, 16 Hz), 4.70 (t, 1H, J=4 Hz), 6.38 (2H, dt, J=16, 8 Hz), 6.7–7.4 (complex, 20H); MS (ES−) m/z 562 ([M−H]+).
Example 5
N,N-bis(4-Benzyloxybenzyl)Lys(Boc)-OMe (cpd 111) and N-(4-Benzyloxybenzyl)Lys(Boc)-OMe
A solution of 594 mg (2.0 mmol) of Lys(Boc)-OMe.HCl, 524 mg (2.25 mmol) of 4-benzyloxybenzyl chloride, 75 mg (0.5 mmol), of NaI, and 0.61 mL (3.5 mol, 452 mg) of DIEA was warmed at 50–70° C. under N2 overnight. The mixture was cooled and partioned between EtOAc and water. The organic layer was washed twice with water, once with brine, and was dried over Na2SO4. The organic solution was concentrated to give 0.83 g of amber oil which was purified by MPLC using a solvent gradient ranging from 15–40% EtOAc/hexanes to give two products. The less polar product (296 mg), cpd 111, was isolated as a pale yellow oil; 1H NMR (CDCl3, 300 MHz) 1.28 (m, 4H), 1.43 (s, 9H), 1.70 (m, 2H), 3.03 (m, 2H), 3.28 (t, 1H, J=7 Hz), 3.40 (d, 2H, J=13.5 Hz), 3.74 (s, 3H), 3.81 (d, 2H, J=13.5 Hz), 5.05 (2, 4H), 6.92 (d, 4H, J=8.5), 7.23 (d, 4H, J=8.5), 7.25–7.5 (complex, 10H); MS (ES+) m/z 653 (MH+).
The more polar product (406 mg), N-(4-benzyloxybenzyl)Lys(Boc)-OMe, was isolated as a white solid; 1H NMR (CDCl3, 300 MHz) 1.4 (, 4H), 1.43 (s, 9H), 1.65 (m, 3H), 3.08 (m, 2H), 3.23 (t, 1H, J=6.5 Hz), 3.54 (d, 1H, J=12.5 Hz), 3.71 (s, 3H), 3.73 (d, 1H, J=12.5 Hz), 5.05 (s, 2H), 6.92 (d, 2H, J=8.5 Hz), 7.23 (d, 2H, J=8.5 Hz), 7.25–7.5 (complex, 5H); MS (ES+) m/z 457 (MH+).
Example 6
N-(4-Benzyloxybenzyl)-N-(3-nitrobenzyl)Lys(Boc)-OMe (cpd 113)
A solution of 374 mg (0.82 mmol) of N-(4-Benzyloxybenzyl)Lys(Boc)-OMe, 221 mg (1.03 mmol) of 4-nitrobenzyl bromide, and 197 L (1.13 mmol, 146 mg) of DIEA was warmed at 50–70° C. for 4 h, then at 40–50° C. overnight. After the addition of 0.2 mL of 1N aqueous HCl, the mixture was partioned between EtOAc and water. The organic layer was washed twice with water, once with brine, and was dried over Na2SO4. The organic solution was concentrated to give 610 mg is of an amber oil which was purified by MPLC 1:3 EtOAc/hexanes to afford 436 mg (90%) cpd 113 as a pale yellow oil; 1H NMR (CDCl3, 300 MHz) 1.35 (m, 4H), 1.42 (s, 9H), 1.75 (broad q, 2H, J=8 Hz), 3.06 (broad q, 2H, J=6 Hz), 3.28 (t, 1H, J=7.5 Hz), 3.48 (d, 1H, J=13.5 Hz), 3.66 (d, 1H, J=14.5 Hz), 3.76 (s, 3H), 3.79 (d, 1H, J=13.5 Hz), 3.97 (d, 1H, J=14.5 Hz), 4.47 (broad s, 1H), 5.05 (s, 2H), 6.93 (d, 2H, J=8.5 Hz), 7.22 (d, 2H, J=8.5 Hz), 7.3–7.5 (complex, 6H), 7.65 (d, 1H, J=7.5 Hz), 8.09 (d, 1H, J=8 Hz), 8.22 (s, 1H); MS (ES+) m/z 592 (MH+).
Example 7
N-(3-Aminobenzyl)-N-(4-benzyloxybenzyl)Lys(Boc)-OMe
A solution of 361 mg (0.61 mmol) of cpd 113 and 835 mg (3.7 mmol) of SnCl2 dihydrate was stirred under N2 at room temperature for 6 h. The slightly cloudy mixture was poured into 200 mL of 5% aqueous Na2CO3 with rapid stirring. The resulting milky suspension was extracted with three 75 mL portions of CH2Cl2 and the combined organic layers were washed with brine and dried over Na2SO4. The extracts were concentrated to give 344 mg of colorless oil which was purified by MPLC using 1:2 EtOAc/hexanes to provide 291 mg of N-(3-aminobenzyl)-N-(4-benzyloxybenzyl)Lys(Boc)-OMe as a yellow oil; 1H NMR (CDCl3, 300 MHz) 1.25 (m, 4H), 1.44 (s, 9H), 1.70 (m, 2H), 3.31 (dd, 1H, J=6, 9 Hz), 3.38 (d, 1H, J=14 Hz), 3.40 (d, 1H, J=13.5 Hz), 3.74 (s, 3H), 3.81 (d, 1H, J=14 Hz), 3.83 (d, 1H, J=13.5 Hz), 4.52 (broad s, 1H), 5.05 (s, 2H), 6.50 (broad d, 1H, J=8 Hz), 6.70 (m, 2H), 6.92 (d, 2H, J=8.5 Hz), 7.08 (t, 1H, J=7.5 Hz), 7.2–7.5 (complex, 7H); MS (ES+) m/z 562 (base, MH+), 506.
Example 8
N-(4-Benzyloxybenzyl)-N-(3-((2-furancarbonyl)amino)benzyl)Lys-OMe (cpd 117)
A solution of 42 mg (0.075 mmol) of N-(3-aminobenzyl)-N-(4-benzyloxybenzyl)Lys(Boc)-OMe and 12 μL (12 mg, 0.15 mmol) of pyridine in 0.5 mL of 1,2-dichloroethane was combined with 8.1 μL (11 mg, 0.083 mmol) and stirred under N2 overnight. EtOAc (3 mL) was added and the solution was washed twice with 2 mL of water and 2 mL of saturated aqueous NaHCO3. The EtOAc solution was filtered through a pad of Na2SO4 and concentrated to give 44 mg of N-(4-benzyloxybenzyl)-N-(3-((2-furancarbonyl)amino)benzyl)Lys(Boc)-OMe; MS (ES+) m/z 356 (MH+). The Boc-protected intermediate was stirred in 2 mL of 50% TFA/CH2Cl2 for 2 h and was concentrated and pumped at high vacuum to provide 66 mg of cpd 117 as the bis-TFA salt; 1H NMR (CD3OD, 300 MHz) 1.55 (m, 2H), 1.65 (m, 2H), 2.10 (m, 2H), 2.93 (t, 2H, J=7 Hz), 3.68 (t, 1H, J=7 Hz), 3.78 (s, 3H), 4.20 (m, 4H), 5.09 (s, 2H), 6.66 (dd, 1H, J=1.5, 3.5 Hz), 7.03 (d, 2H, J=8.5 Hz), 7.1–7.6 (complex, 11H), 7.76 (m, H), 8.07 (m, 1H); MS (ES+) m/z 556 (base, MH+), 360, 197.
Example 9
N,N-bis(3-Nitrobenzyl)Asp(O-t-Bu)-O-t-Bu (cpd 62)
A solution of 0.50 mg (1.77 mmol) of Asp(O-t-Bu)-O-t-Bu.HCl, 1.17 g (5.42 mmol) of 3-nitrobenzyl bromide, and 1.25 mL (0.93 g, 7.2 mmol) of DIEA in 6 mL of DMF was stirred at room temperature under N2 for 24 h and was heated at 70–80° C. overnight. The reaction mixture was partitioned between EtOAc and water and the organic layer was washed twice with water and once with brine. After drying over Na2SO4, the organic solution was concentrated to give 0.86 g of a yellow oil which was purified by MPLC using 1:9 EtOAc/hexanes to afford 0.849 g (93%) cpd 62 as a pale yellow oil; 1H NMR (CDCl3, 300 MHz) 1.43 (s, 9H), 1.57 (s, 9H), 2.59 (dd, 1H, J=7.5, 16 Hz), 2.76 (dd, 1H, J=7, 16 Hz), 3.72 (t, 1H, J=7.5 Hz), 3.78 (d, 2H, J=14 Hz), 3.92 (d, 2H, J=14 Hz), 7.47 (t, 2H, J=8 Hz), 7.67 (d, 2H, J=7.5 Hz), 8.09 (broad d, 2H J=8 Hz), 8.16 (broad s, 2H); MS (ES+) m/z 538 (MNa+), 516 (base, MH+), 460, 404, 237.
Example 10
N,N-bis(3-Aminobenzyl)Asp(O-t-Bu)-O-t-Bu
A solution of 0.644 g (1.25 mmol) of cpd 62 and 2.82 g (12.5 mmol) of SnCl2.2H2O in 12 mL of absolute EtOH was refluxed for 1.5 h. The mixture was cooled and poured into 300 mL of 5% aqueous Na2CO3 with rapid stirring. The cloudy mixture was extracted with three 150 mL portions of CH2Cl2 and the organic extracts were washed with brine and dried over Na2SO4. The CH2Cl2 solution was concentrated to afford 210 mg (37%) of N,N-bis(3-aminobenzyl)Asp(O-t-Bu)-O-t-Bu as a cloudy yellow oil which was used without purification; 1H NMR (CDCl3, 300 MHz) 1.40 (s, 9H), 1.52 (s, 9H), 2.48 (dd, 1H, J=7, 16 Hz), 2.76 (dd, 1H, J=8, 16 Hz), 3.48 (d, 2H, J=14 Hz), 3.55 (m, 1H), 3.73 (d, 2H, J=14 Hz), 6.56 (broad d, 2H J=8 Hz), 6.70 (broad s, 2H), 6.77 (d, 2H, J=7.5 Hz), 7.08 (t, 2H, J=8 Hz); MS (ES+) m/z 478 (MNa+), 456 (base, MH+), 400, 344.
Example 11
N,N-bis(3-(4-Methylbenzoyl)aminobenzyl)Asp(O-t-Bu)-O-t-Bu
To a solution of 109 mg (0.24 mmol) of N,N-bis(3-aminobenzyl)Asp(O-t-Bu)-O-t-Bu, 29 mg (0.24 mmol) of DMAP, 125 μL (93 mg, 0.72 mmol) of DIEA in 1 mL of CH2Cl2 was added 95 μL (111 mg, 0.72 mmol) of 4-methylbenzoyl chloride. The solution was stirred under N2 overnight and was then partitioned between EtOAc and water. The organic layer was washed with saturated aqueous NaHCO3 and brine, dried over Na2SO4, and concentrated to give 177 mg of yellow oil. The crude material was purified by MPLC using a solvent gradient ranging from 20–25% EtOAc/hexanes to provide 87 mg of N,N-bis(3-(4-methylbenzoyl)aminobenzyl)Asp(O-t-Bu)-O-t-Bu as a pale yellow oil; 1H NMR (CDCl3, 300 MHz) 1.36 (s, 9H), 1.55 (s, 9H), 2.35 (s, 6H), 2.53 (dd, 1H, J=6, 16 Hz), 2.76 (dd, 1H, J=9, 16 Hz), 3.69 (d, 2H, J=14), 3.77 (dd, 1H, J=6, 9 Hz), 3.83 (d, 2H, J=14), 7.01 (m, 6H), 7.26 (t, 2H, J=8 Hz), 7.59 (m, 6H), 8.11 (s, 2H), 8.49 (s, 2H); MS (ES+) m/z 714 (MNa+), 692 (base, MH+), 636, 580.
Example 12
N,N-bis(3-(4-Methylbenzoyl)aminobenzyl)Asp-OH (cpd 64)
A solution of 87 mg (0.13 mmol) of N,N-bis(3-(4-methylbenzoyl)amino-benzyl)Asp(O-t-Bu)-O-t-Bu in 1 mL of 50% TFA/CH2Cl2 was stirred overnight. The mixture was concentrated and the residue was dissolved in HOAc and freeze-dried to afford 77 mg cpd 64 as a white solid; 1H NMR (CD3OD, 300 MHz) 2.40 (s, 6H), 2.85 (dd, 1H, J=6, 16.5 Hz), 2.98 (dd, 1H, J=8, 16.5 Hz), 4.02 (d, 2H, J=13.5 Hz), 4.08 (d, 4H, J=13.5 Hz), 4.10 (t, 1H, J=6.5 Hz), 7.22 (m, 6H), 7.34 (t, 2H, J=7.5 Hz), 7.60 (broad d, 2H, J=9 Hz), 7.76 (d, 4H, J=8 Hz), 7.88 (broad s, 2H); MS (ES+) m/z 580 (base, MH+).
Example 13
[N-Cbz-Glu(O-t-Bu)-NHCH2CH2OCH2]2 
To a solution of 1.69 g (5.0 mmol) of N-Cbz-Glu(O-t-Bu)-OH, 0.365 mL (0.371 g, 2.5 mmol) of 1,8-diamino-3,6-dioxaoctane, 0.743 g (5.5 mmol) of HOBT, and 1.05 mL (0.776 g, 6.0 mmol) of DIEA in 15 mL of CH2Cl2 was added 1.05 g (5.5 mmol) of EDCI in one portion. After stirring at room temperature under N2 overnight, the mixture was partitioned between EtOAc and 10% aqueous citric acid. The organic layer was washed with water, saturated NaHCO3, and brine, dried over Na2SO4, and concentrated to give 1.87 g of (N-Cbz-Glu(O-t-Bu)-NHCH2CH2OCH2)2 as a colorless oil; 1H NMR (CD3OD, 300 MHz) 1.43 (s, 18H), 1.85 (m, 2H), 2.05 (m, 2H), 2.31 (t, 4H, J=8 Hz), 3.37 (t, 4H, J=5 Hz), 3.52 (t, 4H, J=5 Hz), 3.58 (s, 4H), 4.15 (m, 2H), 5.09 (dd, 4H, J=12, 16 Hz), 7.30 (m, 10H); MS (ES+) m/z 809 (base, MNa+), 787 (MH+).
Example 14
[Glu(O-t-Bu)-NHCH2CH2OCH2]2 
Ammonium formate (0.78 g, 12.4 mmol) and 0.16 g of 10% palladium on carbon were added to a solution of (N-Cbz-Glu(O-t-Bu)-NHCH2CH2OCH2)2 in 12 mL of MeOH and the resulting suspension was stirred under N2 at room temperature overnight. The mixture was diluted with CH2Cl2 and filtered through a Celite pad. The solids were washed thoroughly with CH2Cl2 and the combined organic filtrates were concentrated to dryness. The residue was partitioned between CH2Cl2 and saturated aqueous NaHCO3, washed with brine, dried over Na2SO4, and concentrated to give 1.13 g of (Glu(O-t-Bu)-NHCH2CH2OCH2)2 as a colorless oil; 1.44 (s, 18H), 1.81 (m, 2H), 2.08 (m, 2H), 2.35 (m, 4H), 3.39 (dd, 2H, J=5, 7.5 Hz), 3.47 (t, 4H, J=5 Hz), 3.58 (t, 4H, J=5 Hz), 7.53 (m, 2H).
Example 15
[N,N-bis(4-Benzyloxybenzyl)Glu(O-t-Bu)-NHCH2CH2OCH2]2 (cpd 245)
A solution of 199 mg (0.384 mmol) of [Glu(O-t-Bu)-NHCH2CH2OCH2]2, 403 mg (1.73 mmol) of 4-benzyloxybenzyl chloride, 30 mg (0.2 mmol) of NaI, and 334 L (248 mg, 1.92 mmol) of DIEA was stirred under N2 at room temperature for several days. The solution was partitioned between EtOAc and water and the organic layer was washed three times with water and once with brine. After drying over Na2SO4, the solution was concentrated to give 528 mg of yellow oil which was purified by MPLC using a solvent gradient ranging from 42–50% EtOAc/hexanes to afford 318 mg (64%) of cpd 245 as a white foam; 1H NMR (CDCl3, 300 MHz) 1.42 (s, 18H), 2.01 (m, 4H), 2.38 (m, 2H), 2.55 (m, 2H), 3.03 (dd, 2H, J=5, 8 Hz), 3.31 (m, 2H), 3.4–3.6 (complex, 18H), 4.99 (s, 8H), 6.89 (d, 8H, J=8.5), 7.1–7.4 (complex, 30H).
Example 16
[N,N-bis(4-Benzyloxybenzyl)GluNHCH2CH2OCH2]2 (cpd 246)
A solution of 219 mg (0.168 mmol) of cpd 245 in 2 mL of 33% TFA/CH2Cl2 was stirred ad room temperature overnight. The mixture was concentrated to give a crude product which was dissolved in HOAc and freeze-dried to afford 251 mg of cpd 246 as an amber oil; 1H NMR (CD3OD, 300 MHz) 2.1–2.6 (complex, 8H), 3.3–3.6 (complex, 8H), 3.57 (s, 4H), 3.78 (m, 2H), 4.25 (broad d, 4H, J=14 Hz), 4.36 (broad d, 4H, J=14 Hz), 5.09 (s, 8H), 7.03 (d, 8H, J=8 Hz), 7.3–7.5 (complex, 28H); MS (ES+) m/z 1192 (MH+), 995, 596, 197 (base).
Example 17
[N-(3-Phenoxybenzyl)Glu(O-t-Bu)-NHCH2CH2OCH2]2 
A solution of 680 mg (0.76 mmol) of [Glu(O-t-Bu)-NHCH2CH2OCH2]2 and 278 μL (317 mg, 1.6 mmol) of 3-phenoxybenzaldehyde in 3 mL of TMOF was stirred overnight at room temperature under N2. The mixture was concentrated and pumped at high vacuum to give a colorless oil which was dissolved in 3 mL of CH2Cl2 and treated with 678 mg (3.2 mmol) of NaBH(OAc)3. After stirring under N2 for 2 days, 50 mL of saturated aqueous NaHCO3 was added and the mixture was extracted with CH2Cl2. The organic layers were combined, dried over Na2SO4, and concentrated and the crude product (1.01 g) was purified by MPLC using a solvent gradient ranging from 2–4% MeOH/CH2Cl2 to afford 490 mg of [N-(3-phenoxybenzyl)Glu(O-t-Bu)-NHCH2CH2OCH2]2 as a colorless oil; 1H NMR (CDCl3, 300 MHz) 1.41 (s, 18H), 1.89 (m, 4H), 2.31 (m, 4H), 3.12 (t, 2H, J=6 Hz), 3.45 (m, 8H), 3.55 (s, 4H), 3.60 (d, 2H, J=13.5 Hz), 3.73 (d, 2H, J=13.5 Hz), 6.86 (dd, 2H, J=1.5, 8 Hz), 7.00 (m, 8H), 7.2–7.4 (complex, 8H); MS (ES+) m/z 883 (MH+), 589, 442, 414, 386 (base), 183.
Example 18
[N-(3-Nitrobenzyl)-N-(3-phenoxybenzyl)-Glu(O-t-Bu)-NHCH2CH2OCH2]2 
DIEA (269 μL, 199 mg, 1.54 mmol), 3-nitrobenzyl bromide (322 mg, 1.49 mmol), and [N-(3-phenoxybenzyl)Glu(O-t-Bu)-NHCH2CH2OCH2]2 (482 mg, 0.546 mmol) were combined in 2 mL of DMF and heated at 60–70° C. under N2 for 2 days. The reaction mixture was cooled and partitioned between 100 mL of EtOAc and water. The organic layer was washed with three times with water and once with brine, dried over Na2SO4, and concentrated to give 661 mg ( ̃100%) of [N-(3-nitrobenzyl)-N-(3-phenoxybenzyl)-Glu(O-t-Bu)-NHCH2CH2OCH2]2 which was used without purification; MS (ES+) m/z 1154 (MH+), 577, 130 (base).
Example 19
[N-(3-Aminobenzyl)-N-(3-phenoxybenzyl)-Glu(O-t-Bu)-NHCH2CH2OCH2]2 
A solution of 661 mg (0.54 mmol) of crude [N-(3-nitrobenzyl)-N-(3-phenoxy-benzyl)-Glu(O-t-Bu)-NHCH2CH2OCH2]2 and 2.71 g (12.0 mmol) of SnCl2. 2H2O in 20 mL of absolute EtOH was refluxed under N2 for 30 min. The cooled solution was poured into 500 mL of 2.5% aqueous Na2CO3 with rapid stirring and the resulting cloudy mixture was extracted thoroughly with EtOAc. The slightly cloudy organic extracts were washed twice with brine, dried over Na2SO4, and concentrated to give 604 mg of yellow oil which was purified by MPLC using 3% MeOH/CH2Cl2 to provide 350 mg (59%) of [N-(3-aminobenzyl)-N-(3-phenoxybenzyl)-Glu(O-t-Bu)-NHCH2CH2OCH2]2 as a pale yellow oil; 1H NMR (CDCl3, 300 MHz) 1.41 (s, 18H), 1.97 (m, 4H), 2.25 (m, 4H), 2.48 (m, 4H), 3.03 (dd, 2H, J=5, 8 Hz), 3.30 (m, 2H), 3.4–3.8 (complex, 24H), 6.47 (d, 2H, J=7.5 Hz), 6.65 (m, 4H), 6.85 (d, 2H, J=9.5 Hz), 6.9–7.15 (complex, 12H), 7.2–7.4 (complex, 8H); MS (ES+) m/z 1094 (MH+), 547 (base).
Example 20
[N-(3-Phenoxybenzyl)-N-(3-(pentanoylamino)benzyl)-Glu-NHCH2CH2OCH2]2 (cpd 247)
Pentanoyl chloride (16 uL, 16 mg, 0.136 mmol) was added dropwise to a solution of 68 mg (0.062 mmol) of [N-(3-aminobenzyl)-N-(3-phenoxybenzyl)-Glu(O-t-Bu)-NHCH2CH2OCH2]2, 20 μL (20 mg, 0.25 mmol) of pyridine in 0.3 mL of 1,2-dichloroethane. The mixture was shaken under N2 overnight and was then partitioned between EtOAc and water. The organic layer was washed with saturated aqueous NaHCO3, dried over Na2SO4, and concentrated to give 77 mg of [N-(3-phenoxybenzyl)-N-(3-(pentanoylamino)benzyl)-Glu(O-t-Bu)-NHCH2CH2OCH2]2; MS (ES+) m/z 1073, 575 (base, MH+/2). The crude product was dissolved in 1 mL of 50% TFA/CH2Cl2 and allows to stand overnight. The solution was concentrated and the resulting oil was dissolved in HOAc and freeze-dried to provide 82 mg of cpd 247; 1H NMR (CD3OD, 300 MHz) 3.98 (t, 6H, J=7.5 Hz), 1.39 (sextet, 4H, J=7.5 Hz), 1.66 (quintet, 4H, J=7.5 Hz), 1.65 (m, 2H), 1.78 (m, 2H), 2.35 (t, 4H, J=7.5 Hz), 2.45 (m, 4H), 3.38 (m, 4H), 3.50 (t, 2H, J=5), 3.57 (m, 4H), 4.10 (broad s, 8H), 6.9–7.25 (complex, 14H), 7.25–7.4 (complex, 10H), 7.71 (s, 2H); MS (ES+) m/z 1150 (MH+), 575 (base).
Example 21
[N-Cbz-Lys(BOC)-NHCH2CH2]3N
A solution of 1.0 g (2.63 mmol) of N-Cbz-Lys(Boc)OH, 0.131 mL (0.128 g, 0.876 mmol) of tris(2-aminoethyl)amine, 0.391 g (2.98 mmol) of HOBt, 0.555 g (2.89 mmol) of EDCI, and 0.55 mL (0.408 g, 3.16 mmol) of DIEA in 5 mL of CH2Cl2 was stirred under N2 at room temperature overnight. The mixture was diluted with EtOAc and washed with 10% aqueous citric acid, saturated aqueous NaHCO3, and brine. The solution was dried over Na2SO4 and concentrated to give 0.872 g of [N-Cbz-Lys(Boc)-NHCH2CH2]3N as an off-white solid; 1H NMR (CD3OD, 300 MHz) 135 (m, 12H), 1.40 (s, 27H), 1.60 (m, 3H), 1.72 (m, 3H), 2.51 (m, 6H), 2.99 (m, 6H), 3.10 (m, 3H), 3.21 (m, 3H), 4.12 (m, 3H), 5.00 (d, 3H, J=12.5 Hz), 5.08 (d, 3H, J=12.5 Hz), 7.29 (m, 15H); MS (ES+) m/z 1243 (base, MH+), 567, 467.
Example 22
[Lys(Boc)-NHCH2CH2]3N
A solution of 0.841 g (0.682 mmol) [N-Cbz-Lys(Boc)-NHCH2CH2]3N, 0.252 g of 10% Pd-C, and 0.774 g (12.3 mmol) of ammonium formate in 10 mL of MeOH was stirred for 5 h at room temperature under N2. The mixture was filtered through a Celite pad, the solids were washed with CH2Cl2, and the reslulting solution was concentrated to dryness. The residue was partitioned between CH2Cl2 and brine; the organic layer was dried over Na2SO4 and concentrated to provide 0.191 g of [Lys(Boc)-NHCH2CH2]3N as an off-white solid; 1H NMR (CD3OD, 300 MHz) 1.40 (s, 27H), 1.45 (m, 12H), 1.75 (m, 6H), 2.62 (m, 6H), 3.01 (m, 6H), 3.28 (m, 6H), 3.64 (m, 3H); MS (ES+) m/z 853 (MNa+), 831 (MH+), 266 (base).
Example 23
[N,N-bis(3-Phenoxybenzyl)Lys(Boc)-NHCH2CH2]3N
A solution of 65 mg (0.078 mmol) of [Lys(Boc)-NHCH2CH2]3N, 120 μL (140 mg, 0.70 mmol) of 3-phenoxybenzaldehyde, and 71 μL (65 mg, 0.70 mmol) of borane-pyridine complexin 3 mL of absolute EtOH was stirred for 4 days at room temperature under N2. The mixture was concentrated to dryness and partitioned between water and CH2Cl2. The organic layer was concentrated to give a yellow oil which was purified by MPLC using 2.5% MeOH/CH2Cl2 to give 78 mg of [N,N-bis(3-phenoxybenzyl)Lys(Boc)-NHCH2CH2]3N as a yellow oil; MS (ES+) m/z 872 (base, [M—C13H12O)/2]+), 611, 443.
Example 24
[N,N-bis(3-Phenoxybenzyl)Lys-NHCH2CH2]3N (cpd 277)
A solution of 78 mg (0.048 mmol) of [N,N-bis(3-phenoxybenzyl)Lys(Boc)-NHCH2CH2]3N in 2 mL of 50% TFA/CH2Cl2 was stirred for 2 h at room temperature. The mixture was diluted with CH2Cl2, washed with water and 5% Na2CO3, and concentrated to give 57 mg of cpd 277 as an off-white foam; 1H NMR (CD3OD, 300 MHz) 1.35 (m, 6H), 1.52 (m, 6H), 1.76 (m, 6H), 2.75 (m, 6H), 3.19 (m, 6H), 3.40 (m, 6H), 3.60 (m, 9H), 3.77 (m, 6H), 6.79 (d, 6H, J=8 Hz), 6.93 (m, 24H), 7.05 (m, 6H), 7.19 (m, 6H), 7.29 (m, 12H); MS (ES+) m/z 813 ([MH2/2]+), 721, 542 (base, [MH/3]+).
Example 25
N,N-bis(3-Phenoxycinnamyl)Ser(t-Bu)-OMe (cpd 290) and N-(3-phenoxycinnamyl)Ser(t-Bu)-OMe (cpd 352)
A solution of 423 mg (2.0 mmol) of H-Ser(t-Bu)OMe.HCl, 1.01 g (3.5 mmol) of 3-phenoxycinnamyl bromide (Jackson, W. P.; Islip, P. J.; Kneen, G.; Pugh, A.; Wates, P. J. J. Med. Chem. 31 1988; 499–500), and 0.87 mL (5.0 mmol, 650 mg) of DIEA in 6 mL of DMF was stirred under N2 at room temperature for 20 h. The mixture was partitioned between EtOAc and water and the organic layer was washed with water and brine. After drying over Na2SO4, the organic solution was concentrated to give 0.98 g of yellow oil. The crude residue was purified by MPLC using a solvent gradient ranging from 10–30% EtOAc/hexanes to give two products. The less polar product (168 mg, 14% based on starting amino acid), N,N-bis(3-phenoxycinnamyl)Ser(t-Bu)-OMe (cpd 290), was isolated as a pale yellow oil; 1H NMR (CDCl3, 300 MHz) 1.15 (s, 9H), 3.35 (dd, 2H, J=7, 14.5 Hz), 3.53 (dd, 2H, J=5.5, 14.5 Hz), 3.6–3.8 (complex, 3H), 3.69 (s, 3H), 6.18 (dt, 2H, J=16, 6.5 Hz), 6.49 (d, 2H, J=16 Hz), 6.86 (dd, 2H, J=2, 8 Hz), 6.9–7.4 (complex, 16H); MS (ES+) m/z 614, 592 (MH+, base), 406, 384, 209.
The more polar product (354 mg, 46% based on starting amino acid), N-(3-phenoxycinnamyl)Ser(t-Bu)-OMe (cpd 352), was isolated as a pale yellow oil; 1H NMR (CDCl3, 300 MHz) 1.15 (s, 9H), 1.98 (broad s, 1H), 3.32 (ddd, 1H, J=1.2, 6.5, 14 Hz), 3.4–3.7 (complex, 4H), 3.72 (s, 3H), 6.21 (dt, 1H, J=16, 6.5 Hz), 6.48 (d, 1H, J=16 Hz), 6.88 (dd, 1H, J=1.5, 8 Hz), 7.0–7.4 (complex, 8H); MS (ES+) m/z 789 (2M+Na+), 384 (MH+, base), 209.
Example 26
N,N-Bis(3-phenoxycinnamyl)Ser-OMe (cpd 299)
N,N-Bis(3-phenoxycinnamyl)Ser(t-Bu)-OMe (cpd 290, 168 mg, 0.284 mmol) was stirred in 3 mL of 50% TFA/CH2Cl2 under N2 overnight. The solvent was removed using a rotary evaporator and the crude residue was partitioned between EtOAc and saturated aqueous NaHCO3. After washing with brine and drying over Na2SO4, the organic layer was concentrated using a rotary evaporator and the crude product (134 mg) was purified by MPLC using 30% EtOAc/hexanes to give 44 mg (29%) of N,N-bis(3-phenoxycinnamyl)Ser-OMe (cpd 299) as a colorless oil; 1H NMR (CDCl3, 300 MHz) 1.6 (broad s, 2H), 3.38 (dd, 2H, J−8, 12 Hz), 3.4–3.9 (complex, 5H), 3.72 (s, 3H), 6.13 (dt, 2H, J=16, 7 Hz), 6.50 (d, 2H, J=16 Hz), 6.8–7.4 (complex, 18H); MS (ES+) m/z 536 (MH+).
Example 27
N,N-Bis(3-phenoxycinnamyl)Ser-OH (cpd 300)
N,N-Bis(3-phenoxycinnamyl)Ser-OMe (cpd 299, 44 mg, 0.082 mmol) was dissolved in 0.2 mL of MeOH and was stirred with 0.090 mL of 1N aqueous NaOH. When TLC analysis revealed that starting material had been consumed, the solvent was removed by rotary evaporation and the residue was lyophilized from acetic acid to give 42 mg (88%) of N,N-bis(3-phenoxycinnamyl)Ser-OH acetate (cpd 300) as a sticky yellow solid; 1H NMR (methanol-d4, 300 MHz) 1.97 (s, 3H), 3.3–4.2 (complex, 7H), 6.80 (d, 2H, J=16 Hz), 6.9–7.4 (complex, 18H); MS (ES+) m/z 522 (MH+), 209.
Example 28
N-(3-Phenoxycinnamyl)Ser-OMe (cpd 346)
N-(3-Phenoxycinnamyl)Ser(t-Bu)-OMe (cpd 352, 268 mg, 0.699 mmol) was stirred in 3 mL of 50% TFA/CH2Cl2 under N2 overnight. The solvent was removed using a rotary evaporator and the crude residue (256 mg) was purified by MPLC using EtOAc to give 137 mg (60%) of N-(3-phenoxycinnamyl)Ser-OMe (cpd 346) as a colorless oil; 1H NMR (CDCl3, 300 MHz) 2.2 (broad s, 2H), 3.36 (dd, 1H, J=6, 14 Hz), 3.4–3.5 (complex, 2H), 3.62 (dd, 1H, J=6.5, 11 Hz), 3.74 (s, 3H), 3.80 (dd, 1H, J=4.5, 11 Hz). 6.19 (dt, 1H, J=16, 6.5 Hz), 6.48 (d, 1H, J=6 Hz), 6.88 (dd, 1H, J=1.5, 8 Hz), 7.0–7.4 (complex, 8H); MS (ES+) m/z 677 (2M+Na+), 350 (M+Na+), 328 (MH+), 209 (base).
Example 29
N-(3-phenoxycinnamyl)Ser-OH (cpd 347)
N-(3-Phenoxycinnamyl)Ser-OMe (cpd 346, 110 mg, 0.336 mmol) was dissolved in 1.5 mL of MeOH and was stirred with 0.50 mL of 1N aqueous NaOH. When TLC analysis revealed that starting material had been consumed, the solvent was removed by rotary evaporation. The residue was dissolved in water and acidified to pH 7–8 with 1N aqueous HCl; the resulting solids were filtered, washed with water, and dried to give 71 mg of white powder. The insoluble powder was dissolved in TFA and, after removal of excess TFA by rotary evaporation, lyophilized from acetic acid to give 82 mg (57%) of N-(3-phenoxycinnamyl)Ser-OH trifluoroacetate (cpd 347) as an amber oil; 1H NMR (methanol-d4, 300 MHz) 3.88 (d, 2H, J=7H), 4.0–4.2 (complex, 3H), 6.27 (dt, 1H, J=16, 6.5), 6.83 (d, 1H, J=16 Hz), 6.9–7.4 (complex, 9H); MS (ES+) m/z 314, (MH+), 209.
Example 30
N-(3-Phenoxycinnamyl)Glu(O-t-Bu)-OH (cpd 337)
A mixture of 249 mg (0.585 mmol) of N-(3-phenoxycinnamyl)Glu(O-t-Bu)-OMe (cpd 334) in 3 mL of MeOH was sonicated to speed dissolution, and the resulting solution was treated with 0.585 mL of 1N aqueous NaOH. After stirring overnight, the MeOH was removed using a rotary evaporator and the residue was dissolved in water. Acidification with 0.64 mL of 1N aqueous HCl produced a 250 mg of solid material that was triturated with Et2O to give 111 mg (46%) of N-(3-phenoxycinnamyl)Glu(O-t-Bu)-OH (cpd 337) as a white solid; 1H NMR (300 MHz, methanol-d4) 1.43 (s, 9H), 1.9–2.2 (complex, 2H), 2.46 (t, 2H, J=7 Hz), 3.57 (dd, 1H, J=5, 7 Hz), 3.78 (dd, 1H, J=7, 13.5 Hz), 3.82 (dd, 1H, J=7, 13.5 Hz), 6.28 (dt, 1H, J=16, 7 Hz), 6.81 (d, 1H, J=16 Hz), 6.9–7.5 (complex, 9H); MS (ES+) m/z 412 (MH+, base), 356, 209. Anal. Calcd for C24H29NO5.0.4H2O: C, 68.55; H, 7.04; N, 3.24. Found: C, 68.89; H, 7.04; N, 3.24.
Example 31
N-(3-Phenoxycinnamyl)Glu-OH (cpd 326)
A mixture of 85 mg (0.21 mmol) of N-(3-phenoxycinnamyl)Glu(O-t-Bu)-OH (cpd 337) in was stirred in 1 mL of 50% TFA/CH2Cl2 for 1 h. After solvent removal using a rotary evaporator, the residue was dissolved in acetic acid and freeze-dried to give 75 mg (76%) of N-(3-phenoxycinnamyl)Glu-OH tifluoroacetate (cpd 326) as a fluffy white solid; 1H NMR (300 MHz, methanol-d4) 2.0–2.4 (complex, 2H), 2.55 (m, 2H), 3.84 (d, 2H, J=7 Hz), 3.96 (dd, 1H, J=5, 7 Hz, 6.24 (dt, 1H, J=16, 7 Hz), 6.84 (d, 1H, J=16 Hz), 6.9–7.4 (complex, 9H); MS (ES+) m/z 356 (MH+), 209 (base).







TABLE 2













 














R1 (amino





cpd
% inh
acid side chain)
R2
R3
W, Q





11
70
Asn, Asp, Gln, Glu
3-PhO

CH═CH


12
59
Cys, Met, Ser, Thr
3-PhO

CH═CH


13
nd
Arg, Gly, His, Pro
3-PhO

CH═CH


14
30
Lys(2-Cl—Cbz),
3-PhO

CH═CH




Phe, Trp, Tyr


15
48
Ala, Ile, Leu, Val
3-PhO

CH═CH


16
nd
Glu, Asp
2,3-benzo

CH═CH


17
nd
Cys, Met
2,3-benzo

CH═CH


18
nd
Ser, Thr
2,3-benzo

CH═CH


19
nd
His, Arg(Mtr)
2,3-benzo

CH═CH


20
nd
Pro, Gly
2,3-benzo

CH═CH


21
nd
Phe, Tyr
2,3-benzo

CH═CH


22
nd
Trp, Lys(2-Cl—Cbz)
2,3-benzo

CH═CH


23
nd
Ile, Ala
2,3-benzo

CH═CH


24
nd
Val, Leu
2,3-benzo

CH═CH


25
nd
Asn, Lys
2,3-benzo

CH═CH


26
nd
Ala, Ile

3,4-benzo
CH═CH


27
nd
Arg(Mtr),

3,4-benzo
CH═CH




Lys(2-Cl—Cbz)


28
nd
Asp, Glu

3,4-benzo
CH═CH


29
nd
Cys, Met

3,4-benzo
CH═CH


30
nd
Gly, Pro

3,4-benzo
CH═CH


31
nd
His, Lys

3,4-benzo
CH═CH


32
nd
Leu, Val

3,4-benzo
CH═CH


33
nd
Lys(2-Cl—Cbz),

3,4-benzo
CH═CH




Phe


34
nd
Ser, Thr

3,4-benzo
CH═CH


35
nd
Trp, Tyr

3,4-benzo
CH═CH
















TABLE 3













 














EPO/EBP-Ig







cpd
% inh @ 50 μM
R1
R2
R9
W, Q
MS MH+





36
nd
CH3
4-CF3
H
CH═CH
458


37
19
H
4-CF3
H
CH═CH
430


38
nd
(CH2)4NH(2-Cl—Cbz)
4-F
H
CH═CH
448


40
nd
CH3
4-F
H
CH═CH
223


41
nd
CH2CO2H
4-F
H
CH═CH
266


42
nd
CH2CH2CO2H
4-F
H
CH═CH
281


43
nd
(CH2)3NHC(═NH)NH2
4-F
H
CH═CH
308


45
nd
PhCH2
4-F
H
CH═CH
299


46
nd
4-HO—PhCH2
4-F
H
CH═CH
315


47
nd
CH2OH
4-F
H
CH═CH
238


48
nd
CH(OH)CH3
4-F
H
CH═CH
253


49
 1
(CH2)3NHC(═NH)NH2
H
H
S
419


50
−6
(CH2)4NH2
H
H
S
391


51
nd
CH(CH3)CH2CH3
H
H
S
376


52
21
CH2CH2CO2H
H
H
S
392


53
14
CH2CO2H
H
H
S
378


54
18
CH3
H
H
S
334


55
 4
CH2CH2CONH2
H
H
S
391


56
nd
(CH2)4NHCbz
H
Me
S
539


57
 0
(CH2)4NHCbz
H
CH2Ph
S
615


58
nd
CH2(indol-3-yl)
H
Me
S
463


59
26
CH2CH2CO2-t-Bu
H
Me
S
462


60
 9
CH2CH2CO2Et
H
Me
S
434


61
14
CH2CH2CO2H
H
Me
S
406
















TABLE 4













 














EPO/EBP-Ig







cpd
% inh @ 50 μM
Ra
R2
R4
R9
MS, MH+





62
nd
t-Bu
NO2
NO2
t-Bu
516


63
20
H
PhCH2NH
PhO
H
511


64
−4
H
4-MePhCONH
4-MePhCONH
H
580


65
−7
H
4-MePhSO2NH
4-MePhSO2NH
H
652


66
−16 
H
3-ClPhCH2NH
PhO
H
546


67
−8
H
3-BrPhCH2NH
PhO
H
590


68
−13 
H
2-FPhCH2NH
PhO
H
529


69
−13 
H
2-MePhCH2NH
PhO
H
525


70
−8
H
4-FPhCH2NH
PhO
H
529


71
−6
H
3-ClPhCH2NH
4-Me—PhO
H
560


72
−14 
H
F5—PhCH2NH
4-Me—PhO
H
615


73
−13 
H
2-FPhCH2NH
4-Me—PhO
H
543


74
−7
H
3-CNPhCH2NH
4-Me—PhO
H
550


75
−2
H
PhCH2NH
4-Me—PhO
H
525
















TABLE 5













 

















EPO/EBP-Ig







MS,


cpd
% inh @ 50 μM
Ra
R2
R3
R4
R5
R9
n
MH+





76
25
t-Bu
PhO
H
PhO
H
t-Bu
1
636


77
52
H
PhO
H
PhO
H
H
1
524


78
nd
H
H
4-MePhCONH
H
BnO
H
2
593


79
nd
H
H
n-BuCONH
H
BnO
H
2
559


80
nd
H
H
2-naphthyl CONH
H
BnO
H
2
629


81
nd
H
H
2-furyl CONH
H
BnO
H
2
569


82
32
H
H
4-MeO—PhCONH
H
BnO
H
2
609


83
18
H
H
HO2C(CH2)3CONH
H
BnO
H
2
589


84
14
H
H
C2F5CONH
H
BnO
H
2
621


85
20
H
H
CF3CONH
H
BnO
H
2
571


86
37
H
H
4-pyridyl-CONH
H
BnO
H
2
580


87
23
H
H
4-MePhSO2NH
H
BnO
H
2
629


88
  10.3
H
H
HO2CCH2(1,1-
H
BnO
H
2
643






cyclopentyl)






CH2CONH


89
22
H
H
PhOCONH
H
BnO
H
2
595


90
29
H
H
4-Ph—PhCONH
H
BnO
H
2
655


91
19
H
H
4-NO2—PhCONH
H
BnO
H
2
624
















TABLE 6













 

















EPO/EBP-Ig







MS,


cpd
% inh @ 50 μM
Ra
R2
R3
R4
R5
n
R9
MH+



















92
20
H
H
H
H
H
2
Me
394


93
20
t-Bu
H
H
H
H
2
Me
450


94
25
Et
H
H
H
H
2
Me
422


95
15
t-Bu
2,3-benzo

2,3-benzo

2
Me
550


96
−5
t-Bu
PhO
H
PhO
H
2
Me
634


97
14
t-Bu
3,4-benzo

3,4-benzo

2
H
536


98
12
t-Bu
H
Ph
H

2
Me
602


99
13
t-Bu
3,4-di-Cl—PhO
H
3,4-di-Cl—PhO
H
2
Me
772


100
34
H
H
Ph
H

2
Me
546


101
32
H
3,4-di-Cl—PhO
H
3,4-di-Cl—PhO
H
2
Me
716


102
5
t-Bu
4-t-Bu—PhO
H
4-t-Bu—PhO
H
2
t-Bu
789


103
17
t-Bu
3-CF3—PhO
H
3-CF3—PhO
H
2
t-Bu
812


104
78
H
4-t-Bu—PhO
H
4-t-Bu—PhO
H
2
H
676


105
70
H
3-CF3—PhO
H
3-CF3—PhO
H
2
H
700


106
20
t-Bu
PhO
H
PhO
H
1
t-Bu
662


107
78
H
PhO
H
PhO
H
2
H
 562*


108
81
H
PhO
H
PhO
H
1
H
550





*[M—H]−













TABLE 7













 
















EPO/










EBP-Ig



% inh






MS,



@ 50






MH


cpd
μM
Ra
R2
R3
R4
R5
R9
+





109
 7
Boc
BnO
H
BnO
H
Me
653


110
54
H
H
BnO
H
BnO
Me
553


111
 5
Boc
H
BnO
H
BnO
Me
653


112
59
H
BnO
H
BnO
H
Me
553


113
24
Boc
H
BnO
NO2
H
Me
592


114
37
H
H
BnO
NO2
H
Me
492


115
35
H
H
BnO
NH2
H
Me
462


116
32
H
H
BnO
n-BuCONH
H
Me
546


117
34
H
H
BnO
2-furylCONH
H
Me
556


118
36
H
H
BnO
4-MePhCONH
H
Me
580


119
34
H
H
BnO
i-Pr—CONH
H
Me
532


120
35
H
H
BnO
4-pyridyl-
H
Me
567







CONH


121
45
H
H
BnO
2-naphthyl-
H
Me
616







CONH


122
nd
Boc
PhCH2NH
H
PhCH2NH
H
Me
651


123
nd
Boc
2-MePhCH2NH
H
2-MePhCH2NH
H
Me
679


124
nd
Boc
4-MeO—
H
4-MeO—
H
Me
711





PhCH2NH

PhCH2NH


125
nd
Boc
3,4-di-MeO—
H
3,4-di-MeO—
H
Me
771





PhCH2NH

PhCH2NH


126
nd
Boc
—NH2
H
—NH2
H
Me
471


127
nd
H
PhCH2NH
H
PhCH2NH
H
Me
551


128
nd
H
2-MePhCH2NH
H
2-MePhCH2NH
H
Me
579


129
nd
H
4-MeO—
H
4-MeO—
H
Me
611





PhCH2NH

PhCH2NH


130
nd
H
3,4-di-MeO—
H
3,4-di-MeO—
H
Me
671





PhCH2NH

PhCH2NH


131
nd
H
PhCH2CH2NH
H
PhCH2CH2NH
H
Me
579


132
nd
HO2CCH2CH2CO
PhCH2NH
H
PhCH2NH
H
Me
651


133
nd
HO2CCH2CH2CO
2-MePhCH2NH
H
2-MePhCH2NH
H
Me
679


134
nd
HO2CCH2CH2CO
4-MeO—
H
4-MeO—
H
Me
711





PhCH2NH

PhCH2NH


135
nd
HO2CCH2CH2CO
3,4-di-MeO—
H
3,4-di-MeO—
H
Me
771





PhCH2NH

PhCH2NH


136
nd
HO2CCH2CH2CO
PhCH2CH2NH
H
PhCH2CH2NH
H
Me
679
















TABLE 8













 















EPO/EBP-Ig









% inh @ 50





MS,


cpd
μM
Ra
R2
R4
R5
R9
MH+





137
nd
H
PhO
PhO
H
Me
551


138
nd
Boc
4-t-Bu—PhO
BnO
H
Me
721


139
nd
H
4-t-Bu—PhO
BnO
H
Me
621


140
nd
H
(CF3CO)2N
BnO
H
H
666


141
nd
H
PhCONH
BnO
H
H
578


142
nd
H
4-pyridyl-CONH
BnO
H
H
579


143
nd
H
(CF3CO)2N
PhO
H
H
652


144
nd
H
PhCONH
PhO
H
H
564


145
nd
H
4-pyridyl-CONH
PhO
H
H
565


146
nd
H
(CF3CO)2N
MeO
MeO
H
620


147
nd
H
PhCONH
MeO
MeO
H
532


148
nd
H
4-pyridyl-CONH
MeO
MeO
H
533


149
nd
H
(CF3CO)2N
H
PhO
H
652


150
nd
H
PhCONH
H
PhO
H
564


151
nd
H
4-pyridyl-CONH
H
PhO
H
565


152
nd
H
PhCONH
H
BnO
H
578


153
nd
H
4-pyridyl-CONH
H
BnO
H
579


154
nd
H
(CF3CO)2N
H
BnO
H
666


155
nd
HO2CCH2CH2CO
4-MeO—PhCONH
PhO
H
H
694


156
nd
HO2CCH2CH2CO
PhCONH
PhO
H
H
664


157
nd
HO2CCH2CH2CO
2-naphthyl-
PhO
H
H
714





CONH


158
nd
HO2CCH2CH2CO
4-Me—PhSO2NH
PhO
H
H
714


159
nd
HO2CCH2CH2CO
4-MeO—PhCONH
2,3-

H
652






benzo


160
nd
HO2CCH2CH2CO
PhCONH
2,3-

H
622






benzo


161
nd
HO2CCH2CH2CO
2-naphthyl-
2,3-

H
672





CONH
benzo


162
nd
HO2CCH2CH2CO
4-Me—PhSO2NH
2,3-

H
672






benzo


163
nd
HO2CCH2CH2CO
4-MeO—PhCONH
H
F
H
620


164
nd
HO2CCH2CH2CO
PhCONH
H
F
H
590


165
nd
HO2CCH2CH2CO
2-naphthyl-
H
F
H
640





CONH


166
nd
HO2CCH2CH2CO
4-Me—PhSO2NH
H
F
H
640


167
nd
HO2CCH2CH2CO
4-MeO—PhCONH
BnO
H
H
708


168
nd
HO2CCH2CH2CO
PhCONH
BnO
H
H
678


169
nd
HO2CCH2CH2CO
2-naphthyl-
BnO
H
H
728





CONH


170
nd
HO2CCH2CH2CO
4-Me—PhSO2NH
BnO
H
H
728
















TABLE 9













 
















EPO/










EBP-



Ig



% inh






MS,



@ 50






MH


cpd
μM
Ra
R2
R3
R4
R5
R9
+





171
nb
Cbz
H
H
H
H
Me
527


172
15
Cbz
H
H
H
H
H
513


173
 5
Cbz
H
H
H
H
t-
569









Bu


174
23
Cbz
H
MeO
H
MeO
Me
587


175
 1
Cbz

3,4-

3,4-
Me
627






benzo

benzo


176
−4
Cbz
PhO
H
PhO
H
Me
711


177
nd
Cbz
2,3-benzo

2,3-benzo

Me
627


178
36
Boc
H
NO2
H
NO2
Me
583


179
30
Boc
H
NO2
H
NO2
H
569


180
−4
Boc
PhO
H
PhO
H
Me
677


181
−9
Boc
4-t-Bu—PhO
H
4-t-Bu—PhO
H
Me
790


182
18
H
4-t-Bu—PhO
H
4-t-Bu—PhO
H
Me
689


183
36
Boc
NO2
H
NO2
H
Me
583


184
53
H
NO2
H
NO2
H
Me
483


185
29
H
NH2
H
NH2
H
Me
423


186
nd
H
n-Bu—CONH
H
n-Bu—CONH
H
Me
591


187
nd
H
2-furyl-CONH
H
2-furyl-CONH
H
Me
611


188
nd
H
PhCONH
H
PhCONH
H
Me
631


189
nd
H
4-Me—PhCONH
H
4-Me—PhCONH
H
Me
659


190
nd
H
4-NO2—PhCONH
H
4-NO2—PhCONH
H
Me
721


191
nd
H
4-Me—PhSO2NH
H
4-Me—PhSO2NH
H
Me
731


192
nd
H
Cbz—NH
H
Cbz—NH
H
Me
691


193
nd
H
4-Br—PhCONH
H
4-Br—PhCO
H
Me
789


194
nd
H
2-MeO—PhCONH
H
2-MeO—PhCONH
H
Me
691


195
nd
H
3-MeO—PhCONH
H
3-MeO—PhCONH
H
Me
691


196
nd
H
4-MeO—PhCONH
H
4-MeO—PhCONH
H
Me
691


197
nd
H
CH3CH═CHCONH
H
CH3CH═CHCONH
H
Me
559


198
nd
H
C2F5CONH
H
C2F5CONH
H
Me
715


199
nd
H
2-naphthyl-
H
2-naphthyl-
H
Me
731





CONH

CONH


200
nd
H
EtO2CCH2CH2CONH
H
EtO2CCH2CH2CONH
H
Me
679


201
nd
H
CF3CONH
H
CF3CONH
H
Me
615


202
nd
H
MeSO2NH
H
MeSO2NH
H
Me
579
















TABLE 10













 
















EPO/










EBP-Ig






MS,



% inh @






MH


cpd
50 μM
Ra
R2
R3
R4
R5
Z
+





203
37
Boc
H
H
H
H
4-(MeCOCH2CH2)—PhNH
640


204
−6
H
H
H
H
H
4-(MeCOCH2CH2)—PhNH
540


205
26
H
H
H
H
H
n-Bu—NH
434


206
17
2-MeO—PhCO
H
H
H
H
n-Bu—NH
568


207
20
4-MeO—PhCO
H
H
H
H
n-Bu—NH
568


208
22
PhCO
H
H
H
H
n-Bu—NH
538


209
25
2-MeO—PhCO
H
H
H
H
n-Bu—NH
568


210
nd
Boc
H
H
H
H
4-MeO—PhCH2CH2NH
612


211
62
H
H
H
H
H
4-MeO—PhCH2CH2NH
512


212
−10 
H
H
H
H
H
n-Pr—NH
420


214
nd
Boc
H
H
H
H
3,4-di-MeO—
642









PhCH2CH2NH


215
nd
Boc
H
H
H
H
3-MeO—PhCH2CH2NH
612


216
10
Boc
H
H
H
H
4-(PhCH═CHCH2O)—
700









PhCH2NH


217
nd
Boc
H
H
H
H
4-HO—PhCH2NH
584


218
nd
Boc
H
H
H
H
EtNH
506


219
nd
Boc
H
H
H
H
MeNH
492


220
45
H
H
H
H
H
4-(PhCH═CHCH2O)—
600









PhCH2NH


221
48
H
H
H
H
H
3,4-di-MeO—
542









PhCH2CH2NH


222
56
H
H
H
H
H
3-MeO—PhCH2CH2NH
512


223
nd
Boc
H
H
H
H
2-MeO—PhCH2CH2NH
612


224
51
H
H
H
H
H
2-MeO—PhCH2CH2NH
512


225
10
Boc
PhO
H
PhO
H
4-MeO—PhCH2CH2NH
797


226
nd
Boc
H
H
H
H
PhCH2CH2NH
582


227
48
H
H
H
H
H
PbCH2CH2NH
482


228
21
PhNHCO
PhO
H
PhO
H
4-MeO—PhCH2CH2NH
816


229
22
4-PhO—
H
H
H
H
4-MeO—PhCH2CH2NH
723




PhNHCO


230
42
3,4-di-Cl—
H
H
H
H
4-MeO—PhCH2CH2NH
700




PhNHCO


231
36
4-EtO2C—
H
H
H
H
4-MeO—PhCH2CH2NH
703




PhNHCO


232
14
4-PhO—
PhO
H
PhO
H
4-MeO—PhCH2CH2NH
908




PhNHCO


233
18
H
H
NO2
H
NO2
3-MeO—PhCH2CH2NH
602


234
nd
Boc
H
H
H
H
PbCH2NH
568


235
49
H
H
H
H
H
PhCH2NH
468


236
nd
Boc
H
Ph
H
Ph
4-MeO—PhCH2CH2NH
765


237
55
HO2CCH2CH2CO
H
H
H
H
3-MeO—PhCH2CH2NH
612


238
39
H
H
Ph
H
Ph
4-MeO—PhCH2CH2NH
664


239
46
H
PhO
H
PhO
H
PhCH2CH2NH
666


240
nd
HO2CCH2CH2CH2CO
PhO
H
PhO
H
PhCH2CH2NH
780


285
40
H
H
H
H
H
4-(NH2CO)piperidin-1-yl
489
















TABLE 11













 

















EPO/











EBP-Ig



% inh @







MS,


cpd
50 μM
Ra
R2
R3
R4
R5
Z
r
[MH2/2]+



















241
2
t-Bu
H
BnO
H
BnO
NH(CH2)3O(CH2)4O(CH2)3NH
1
666


242
1
t-Bu
H
BnO
H
BnO
NH(CH2)3O(CH2CH2O)2(CH2)3NH
1
674


243
75
H
H
BnO
H
BnO
NH(CH2)3O(CH2)4O(CH2)3NH
1
610


244
66
H
H
BnO
H
BnO
NH(CH2)3O(CH2CH2O)2(CH2)3NH
1
618


245
0
t-Bu
H
BnO
H
BnO
NH(CH2)2O(CH2)2O(CH2)2NH
2
652


246
79
H
H
BnO
H
BnO
NH(CH2)2O(CH2)2O(CH2)2NH
2
596


247
47
H
n-Bu—
H
PhO
H
NH(CH2)2O(CH2)2O(CH2)2NH
2
575





CONH


248
56
H
2-furyl-
H
PhO
H
NH(CH2)2O(CH2)2O(CH2)2NH
2
585





CONH


249
72
H
4-Me—
H
PhO
H
NH(CH2)2O(CH2)2O(CH2)2NH
2
609





PhCONH


250
78
H
4-Me—
H
PhO
H
NH(CH2)2O(CH2)2O(CH2)2NH
2
645





PhSO2NH
















TABLE 12













 















EPO/EBP-Ig









% inh @ 50





MS,


cpd
μM
Ra
R2
R4
Z
r
[MH2/2]+





251
49
H
H
H
NH(CH2)2O(CH2)2O(CH2)2NH
2
436


252
−4
t-Bu
4-t-Bu—
4-t-Bu—PhO
NH(CH2)3O(CH2)4O(CH2)3NH
1
803





PhO


253
−5
t-Bu
4-t-Bu—
4-t-Bu—PhO
NH(CH2)3O(CH2CH2O)2(CH2)3NH
1
811





PhO


254
−9
t-Bu
4-t-Bu—
4-t-Bu—PhO
NH(CH2)10NH
1
787





PhO


255
 0
t-Bu
4-t-Bu—
4-t-Bu—PhO
NH(CH2)12NH
1
801





PhO


256
10
t-Bu
4-t-Bu—
4-t-Bu—PhO
NH(CH2)2O(CH2)2O(CH2)2NH
1
789





PhO
















TABLE 13













 












EPO/EBP-Ig


MS,


cpd
% inh @ 50 μM
Ra
Z
[MH2/2]+














257
−26
Boc
NH(CH2)3O(CH2CH2O)2(CH2)3NH
731


258
−24
Boc
NH(CH2)3O(CH2)4O(CH2)3NH
723


259
−13
Boc
NH(CH2)12NH
721


260
−12
Boc
NH(CH2)2O(CH2)2O(CH2)2NH
695


261
51
H
NH(CH2)2O(CH2)2O(CH2)2NH
595


262
93
HO2CCH2CH2CO
NH(CH2)2O(CH2)2O(CH2)2NH
695


263
88
HO2C(CH2)3CO
NH(CH2)2O(CH2)2O(CH2)2NH
709


264
89
HO2CCH2CMe2CH2CO
NH(CH2)2O(CH2)2O(CH2)2NH
737


265
65
HO2CCH2CH2CO
NH(CH2)3O(CH2)4O(CH2)3NH
723


266
82
HO2C(CH2)3CO
NH(CH2)3O(CH2)4O(CH2)3NH
737


267
83
HO2CCH2CMe2CH2CO
NH(CH2)3O(CH2)4O(CH2)3NH
765


268
40
HO2CCH2CMe2CH2CO
NH(CH2)12NH
764


269
55
HO2CCH2CH2CH2CO
NH(CH2)12NH
735


270
56
HO2CCH2CH2CO
NH(CH2)12NH
721


271
77
HO2CCH2CH2CO
NH(CH2)3O(CH2CH2O)2(CH2)3NH
731


272
78
HO2CCH2CH2CH2CO
NH(CH2)3O(CH2CH2O)2(CH2)3NH
745
















TABLE 14













 















EPO/









EBP-Ig





MS,



% inh @





[MH2/2]


cpd
50 μM
Ra
R2
R4
Z
n
+





273
nd
HO2CCH2CH2CO
4-Me—PhO
4-Me—PhO
NH(CH2)2O(CH2)2O(CH2)2NH
2
695


274
nd
HO2CCH2CH2CO
PhO
PhO
NH(CH2)2O(CH2)2O(CH2)2NH
2
667


275
nd
HO2CCH2CH2CO
4-MeO—
4-MeO—
NH(CH2)2O(CH2)2O(CH2)2NH
2
727





PhO
PhO


276
nd
HO2CCH2CH2CO
4-t-Bu—
4-t-Bu—
NH(CH2)2O(CH2)2O(CH2)2NH
2
780





PhO
PhO


277
nd
H
PhO
PhO
(NHCH2CH2)3N
3
813


278
nd
H
4-Me—PhO
4-Me—PhO
(NHCH2CH2)3N
3
855


279
nd
H
4-MeO—
4-MeO—
(NHCH2CH2)3N
3
903





PhO
PhO


280
nd
HO2CCH2CH2CO
4-MeO—
4-MeO—
(NHCH2CH2)3N
3
1053 





PhO
PhO


281
nd
HO2CCH2CH2CO
4-Me—PhO
4-Me—PhO
(NHCH2CH2)3N
3
1005 


282
nd
HO2CCH2CH2CO
PhO
PhO
(NHCH2CH2)3N
3
963


283
nd
Boc
PhO
PhO
NH(CH2)3NMe(CH2)3NH
2
666


284
nd
Boc
4-Me—PhO
4-Me—PhO
NH(CH2)3NMe(CH2)3NH
2
694
















TABLE 15













 













EPO/EBP-Ig






cpd
% inh @ 50 μM
R1
R2
R3
MS, MH+





285
−28 
Me
H
H
473


286
46
H
BnO
H
565


287
36
H
4-Me—PhO
H
565


288
27
H
4-tBu—PhO
H
607


289
20
H
H
PhO
551
















TABLE 16A













 













EPO/EBP-Ig






cpd
% inh @ 50 μM
R1
R2
R3
R4





290
 0
Me
PhO
H
t-Bu


291
17
Me
H
Ph
t-Bu


292
11
Me
3-CF3—C6H4O
H
t-Bu


293
14
Me
3,4-Cl2—C6H3O
H
t-Bu


294
 9
Me
4-t-Bu—C6H4O
H
t-Bu


295
10
Me
H
Ph
H


296
 0
Me
3,4-Cl2—C6H3O
H
H


297
 0
Me
3-CF3—C6H4O
H
H


298
 1
Me
4-t-Bu—C6H4O
H
H


299
nd
Me
PhO
H
H


300
57
H
PhO
H
H


301
25
H
H
Ph
t-Bu


302
30
H
3,4-Cl2—C6H3O
H
t-Bu


303
21
H
3-CF3—C6H4O
H
t-Bu


304
19
H
4-t-Bu—C6H4O
H
t-Bu


305
48
H
H
Ph
H


306
21
Me
H
H
t-Bu


307
25
H
3,4-Cl2—C6H3O
H
H


308
25
H
3-CF3—C6H4O
H
H


309
13
H
4-t-Bu—C6H4O
H
H


310
34
Me
H
H
H




















TABLE 16B





cpd
MPLC solvent
appearance
empirical formula
MS, MH+



















290
10–30%
pale yellow oil
C38H41NO5
592



EtOAc/hex


291
1:5 EtOAc/hex
yellow oil
C38H41NO3
560


292
1:5 EtOAc/hex
yellow oil
C40H39F6NO5
728


293
1:5 EtOAc/hex
yellow oil
C38H37Cl4NO5
728


294
1:5 EtOAc/hex
yellow oil
C46H57NO5
704


295

off-white solid
C34H33NO3/1
504





C2H4O2


296

amber solid
C34H29Cl4NO5/1
672





C2H4O2


297

amber oil
C36H31F6NO5/1
672





C2H4O2


298

off-white solid
C42H49NO5/1
648





C2H4O2


299
30% EtOAc/hex
colorless oil
C34H33NO5
536


300

sticky yellow solid
C33H31NO5/1
522





C2H4O2


301

yellow solid
C37H39NO3
546


302

amber oil
C37H35Cl4NO5
714


303

amber oil
C39H37F6NO5
714


304

amber oil
C45H55NO5
690


305

amber solid
C33H31NO2/1
490





C2HF3O2


306

light-yellow oil
C26H33NO3/0.25
408





H2O


307

amber solid
C33H27Cl4NO5/1
658





C2HF3O2


308

amber oil
C35H29F6NO5/1
658





C2HF3O2


309

off-white solid
C41H47NO5/1
634





C2HF3O2


310

light yellow oil
C22H25NO3/1
352





C2H4O2
















TABLE 17A













 














EPO/EBP-Ig






cpd
% inh @ 50 μM
R1
R2
R3

















311
5.3
t-Bu
PhO
H



312
45
H
PhO
H





















TABLE 17B





cpd
MPLC solvent
appearance
empirical formula
MS, MH+







311
10% EtOAc/hex
pale yellow oil
C36H37NO4
548


312

sticky brown
C32H29NO4/1
492




solid
C2HF3O2
















TABLE 18A













 













EPO/







EBP-Ig,



% inh @


cpd
50 μM
R1
R2
R3
R4





313
28
H
H
CF3
(CH2)4NH(2-Cl—







Cbz)


314
12
Me
H
CO2H
(CH2)4NH2


315
nd
Me
H
NO2
(CH2)4NHBoc


316
20
Me
OPh
H
(CH2)4NHBoc


317
13
Me
4-t-Bu—
H
(CH2)4NHBoc





C6H4O


318
14
Me
H
H
(CH2)4NHCbz


319
nd
Me
H
H
(CH2)4NHCbz


320
17
Me
H
OMe
(CH2)4NHCbz


321
42
Me
CO2Me
H
(CH2)4NHCbz


322
nd
Me
H
2,3-benzo
(CH2)4NHCbz


323
 6
Me
H
CO2H
(CH2)4NHCbz


324
nd
Me
H
CO2Me
nd




















TABLE 18B





cpd
MPLC solvent
appearance
empirical formula
MS, MH+



















313

yellow oil
C25H28ClF3N2O4\1
499





C2HF3O2


314

yellow oil
C17H24N2O4
321


315
30% EtOAc/hex
dark yellow gum
C21H31N3O6
422


316
20–50% EtOAc/hex
pale yellow oil
C27H36N2O5
469


317

pale yellow oil
C31H44N2O5
525


318

gum
C24H30N2O4
411


319

pale yellow oil
C24H30N2O4
411


320
2% MeOH/CH2Cl2
yellow oil
C25H32N2O5
441


321

yellow oil
C26H32N2O6\1
469





C2H4O2


322
25–50% EtOAc/hex
clear residue
C28H32N2O4
461


323

yellow oil
C25H30N2O6
455


324

yellow oil
C26H32N2O6
469
















TABLE 19A













 













EPO/







EBP-Ig



% inh @


cpd
50 μM
R1
R2
R3
R4





325
 6
Me
OPh
H
CH2CH2CO2H


326
 0
H
OPh
H
CH2CH2CO2H


327
11
Me
H
Ph
CH2CH2CO2H


328
33
Me
3,4-Cl2—C6H3O
H
CH2CH2CO2H


329
13
H
H
Ph
CH2CH2CO2H


330
12
H
3-CF3—C6H4O
H
CH2CH2CO2H


331
18
H
4-t-Bu—C6H4O
H
CH2CH2CO2H


332
17
H
3,4-Cl2—C6H3O
H
CH2CH2CO2H


333
16
Me
3,4-benzo

CH2CH2CO2-t-Bu


334
 6
Me
OPh
H
CH2CH2CO2-t-Bu


335
25
Me
H
Ph
CH2CH2CO2-t-Bu


336
32
Me
3,4-Cl2—C6H3O
H
CH2CH2CO2-t-Bu


337
 0
H
OPh
H
CH2CH2CO2-t-Bu


338
23
t-Bu
3-CF3—C6H4O
H
CH2CH2CO2-t-Bu


339
10
t-Bu
4-t-Bu—C6H4O
H
CH2CH2CO2-t-Bu


340
14
H
H
Ph
CH2CH2CO2-t-Bu


341
19
H
3,4-Cl2—C6H3O
H
CH2CH2CO2-t-Bu




















TABLE 19B





cpd
MPLC solvent
appearance
empirical formula
MS, MH+



















325

off-white solid
C21H23NO5\1
370





C2F3HO2


326

fluffy white
C20H21NO5\1
356




solid
C2HF3O2


327

off-white solid
C21H23NO4\1
354





C2F3HO2


328

amber oil
C21H21Cl2NO5\1
438





C2F3HO2


329

amber solid
C20H21NO4\1
340





C2HF3O2


330

amber oil
C21H20F3NO5\1
424





C2HF3O2


331

amber oil
C24H29NO5\1
412





C2HF3O2


332

amber oil
C20H19CL2NO5\1
424





C2HF3O2
424


333
10–25% EtOAc/hex
yellow oil
C23H29NO4
384


334
10–30% EtOAc/hex
pale yellow oil
C25H31NO5
426


335
1:5 EtOAc/hex
yellow oil
C25H31NO4
410


336
1:5 EtOAc/hex
yellow oil
C25H29C12NO5
494


337

white powder
C24H29NO5\0.4
412





H2O


338
1:5 EtOAc/hex
yellow oil
C29H36F3NO5
536


339
1:5 EtOAc/hex
yellow oil
C32H45NO5
524


340

yellow solid
C24H29NO4
396


341

white solid
C24H27C12NO5
480
















TABLE 20A













 













EPO/EBP-Ig






cpd
% inh @ 50 μM
R1
R2
R3
R4





342
 0
Me
H
Ph
CH2OH


343
37
Me
4-t-Bu—C6H4O
H
CH2OH


344
 4
Me
3-CF3—C6H4O
H
CH2OH


345
40
Me
3,4-Cl2—C6H3O
H
CH2OH


346
28
Me
OPh
H
CH2OH


347
23
H
OPh
H
CH2OH


348
21
H
H
Ph
CH2OH


349
23
H
3,4-Cl2—C6H3O
H
CH2OH


350
23
H
3-CF3—C6H4O
H
CH2OH


351
29
H
4-t-Bu—C6H4O
H
CH2OH


352
 8
Me
OPh
H
CH2O-t-Bu


353
24
Me
H
Ph
CH2O-t-Bu


354
31
Me
3,4-Cl2—C6H3O
H
CH2O-t-Bu


355
22
Me
3-CF3—C6H4O
H
CH2O-t-Bu


356
23
Me
4-t-Bu—C6H4O
H
CH2O-t-Bu


357
12
H
3-CF3—C6H4O
H
CH2O-t-Bu




















TABLE 20B





cpd
MPLC solvent
appearance
empirical formula
MS, MH+



















342

off-white solid
C19H21NO3\1
312





C2F3HO2


343

amber oil
C23H29NO4\1
384





C2F3HO2


344

amber oil
C20H20F3NO4\1
396





C2F3HO2


345

amber oil
C19H19Cl2NO4\1
396





C2F3HO2


346
EtOAc
pale yellow oil
C19H21NO4
328


347

amber oil
C18H19NO4\1
314





C2HF3O2


348

yellow solid
C18H19NO3\1
298





C2HF3O2


349

amber oil
C18H17Cl2NO4\1
382





C2HF3O2


350

amber oil
C19H18F3NO4\1
382





C2HF3O2


351

amber oil
C22H27NO4\1
370





C2HF3O2


352
10–30% EtOAc/hex
pale yellow oil
C23H29NO4
384


353
20% EtOAc/hex
off-white solid
C23H29NO3
368


354
20% EtOAc/hex
yellow oil
C23H27Cl2NO4
452


355
20% EtOAc/hex
yellow oil
C24H28F3NO4
452


356
20% EtOAc/hex
yellow oil
C27H37NO4
440


357

white solid
C23H26F3NO4
438
















TABLE 21A













 













EPO/EBP-Ig






cpd
% inh @ 50 μM
R1
R2
R3
R4















358
0
H
H
CF3
(s)-CH(OH)CH3


359
25
Me
CO2Me
H
(s)-CH(OMe)CH3


360
18
Me
H
H
Bn


361
24
Me
CO2Me
H
Bn


362
0
H
H
CF3
CH2(4-HOC6H4)


363
33
Me
CO2Me
H
CH2(4-MeOC6H4)


364
16
Me
H
H
CH2(indol-3-yl)


365
0
H
H
CF3
CH2CH2SMe


366
38
Me
CO2Me
H
CH2CO2Me


367
0
H
H
CF3
CH2CONH2


368
40
Me
CO2Me
H
CH2SBn


369
12
H
H
CF3
i-Bu


370
0
H
H
CF3
i-Pr


371
16
Me
CO2Me
H
i-Pr


372
0
Me
H
H
Me




















TABLE 21B





cpd
MPLC solvent
appearance
empirical formula
MS, MH+



















358

amber oil
C14H16F3NO3\1
304





C2HF3O2


359

amber oil
C17H23NO5\1
322





C2H4O2


360
20% EtOAc/hex
light-yellow oil
C19H21NO2
296


361

amber oil
C22H25NO5\1
354





C2H4O2


362

amber oil
C19H18F3NO3\1
366





C2HF3O2


363

amber oil
C22H25NO5\1
384





C2H4O2


364
1:2 EtOAc/hex
tan solid
C21H22N2O2
335


365

amber oil
C15H18F3NO2S\1
334





C2HF3O2


366

amber oil
C17H21NO6\1
336





C2H4O2


367

amber oil
C14H15F3N2O3\1
317





C2HF3O2


368

amber oil
C22H25NO4S\1
400





C2H4O2


369

amber oil
C17H22F3NO2\1
316





C2HF3O2


370

amber oil
C15H18F3NO2\1
302





C2HF3O2


371

amber oil
C17H23NO4\1
306





C2H4O2


372
20% EtOAc/hex
yellow oil
C13H17NO2\0.10
220





C4H8O2

## Claims

1. A compound of the formula:




wherein:
R1 is the side chain of a natural or unnatural α-amino acid, where if said side chain contains a protectable group, that group may be protected with a member of the group consisting of succinyl, glutaryl, 3,3-dimethylglutaryl, C1-5alkyl, C1-5alkoxycarbonyl, acetyl, N-(9-fluorenylmethoxycarbonyl), trifluoroacetyl, omega-carboxyC1-5alkylcarbonyl, t-butoxycarbonyl, benzyl, benzyloxycarbonyl, 2-chlorobenzyloxycarbonyl, phenylsulfonyl, ureido, t-butyl, cinnamoyl, trityl, 4-methyltrityl, 1-(4,4-dimethyl-2,6-dioxocyclohexylidene)ethyl, tosyl, 4-methoxy-2,3,6-trimethylbenzenesulfonyl, phenylureido, and substituted phenylureido(where the phenyl substituents are phenoxy, halo, C1-5alkoxycarbonyl);
R2 and R3 may be taken together to form a six-membered aromatic ring which is fused to the depicted ring, or are independently selected from the group consisting of hydrogen, C1-5alkyl, C1-5alkoxy, hydroxy, halo, trifluoromethyl, nitro, amino, phenyl, phenoxy, phenyl C1-5alkyl, phenyl C1-5alkoxy,
substituted phenyl (where the substituents are selected from C1-5alkyl, C1-5alkoxy, hydroxy, halo, trifluoromethyl, nitro, cyano, and amino),
substituted phenylC1-5alkyl(where the substituents are selected from C1-5alkyl, C1-5alkoxy, hydroxy, halo, trifluoromethyl, nitro, cyano, and amino), and
substituted amino (where the substituents are selected from one or more members of the group consisting of C1-5alkyl, halosubstitutedC1-5alkyl, C1-5alknyl, C1-5alkenyl, phenyl, phenylC1-5alkyl, C1-5alkylcarbonyl, halosubstituted, C1-5alkylcarbonyl, carboxyC1-5alkyl, C1-5alkoxyC1-5alkyl, cinnamoyl, naphthylcarbonyl, furylcarbonyl, pyridylcarbonyl, C1-5alkylsulfonyl, phenylcarbonyl, phenylC1-5alkylcarbonyl, phenylsulfonyl, phenylC1-5alkylsulfonyl substituted phenylcarbonyl, substituted phenylC1-5alkylcarbonyl, substituted phenylsulfonyl, substituted phenylC1-5alkylsulfonyl, substituted phenyl, and substituted phenylC1-5alkyl[where the aromatic phenyl, phenylC1-5alkyl, phenylcarbonyl, phenylC1-5alkylcarbonyl, phenylsulfonyl, and phenylC1-5alkylsulfonyl substituents are independently selected from one to five members of the group consisting of C1-5alkyl, C1-5alkoxy, hydroxy, halogen, trifluoromethyl, nitro, cyano, and amino]); and
W and Q is CH═CH.

2. A pharmaceutical composition comprising the compound of claim 1.

3. A pharmaceutical composition comprising an active drug component and the compound of claim 1.

4. The composition of claim 3 wherein said active drug component is combined with an oral, non-toxic pharmaceutically acceptable inert carrier.

5. The composition of claim 4 wherein said carrier is ethanol, glycerol, or water.

6. The composition of claim 3 further comprising binders, lubricants, disintegrating agents, or coloring agents.

7. The composition of claim 6 wherein said binders are selected from the group consisting of starch, gelatin, natural sugars, corn sweeteners, natural and synthetic gums carboxymethylcellulose, polyethylene glycol, and waxes.

8. The composition of claim 6 wherein said lubricants are selected from the group consisting of sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, and sodium chloride.

9. The composition of claim 6 wherein said disintegrating agents are selected from the group consisting of starch, methyl cellulose, agar, bentonite, and xanthan gum.

10. The composition of claim 3 contained in a topical administration.

11. The composition of claim 10 wherein said active drug component can be admixed with carrier materials selected from the group consisting of alcohols, aloe vera gel, allontoin, glycerine, vitamin A and E oils, mineral oil, PPG2 myristyl propionate.

12. An oral composition comprising an EPO receptor modulating compound of claim 1 having an active drug component being combined with an oral, non-toxic pharmaceutically acceptable inert carrier.

13. The composition of claim 12 wherein said carrier is ethanol, glycerol, or water.

14. The composition of claim 12 further comprising binders, lubricants, disintegrating agents, or coloring agents.

15. The composition of claim 14 wherein said binders are selected from the group consisting of starch, gelatin, natural sugars, corn sweeteners, natural and synthetic gums carboxymethylcellulose, polyethylene glycol, and waxes.

16. The composition of claim 14 wherein said lubricants are selected from the group consisting of sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, and sodium chloride.

17. The composition of claim 14 wherein said disintegrating agents are selected from the group consisting of starch, methyl cellulose, agar, bentonite, and xanthan gum.

18. A compound of the formula:




wherein:
R1 is H or the side chain of a natural or unnatural α-amino acid, where if said side chain contains a protectable group, that group may be protected with a member of the group consisting of succinyl, glutaryl, 3,3-dimethylglutaryl, C1-5alkyl, C1-5alkoxycarbonyl, acetyl, N-(9-fluorenylmethoxycarbonyl), trifluoroacetyl, omega-carboxyC1-5alkylcarbonyl, t-butoxycarbonyl, benzyl, benzyloxycarbonyl, 2-chlorobenzyloxycarbonyl, phenylsulfonyl, ureido, t-butyl, cinnamoyl, trityl, 4-methyltrityl, 1-(4,4-dimethyl-2,6-dioxocyclohexylidene)ethyl, tosyl, 4-methoxy-2,3,6-trimethylbenzenesulfonyl, phenylureido, and substituted phenylureido(where the phenyl substituents are phenoxy, halo, C1-5alkoxycarbonyl);
R2 is H, halo, or trifluoromethyl;
W is CH═CH or S; and
R9 is H, C1-5alkyl, or phenylC1-5alkyl.

19. A pharmaceutical composition comprising the compound of claim 18.

20. A pharmaceutical composition comprising an active drug component and the compound of claim 18.

21. The composition of claim 20 wherein said active drug component is combined with an oral, non-toxic pharmaceutically acceptable inert carrier.

22. The composition of claim 21 wherein said carrier is ethanol, glycerol, or water.

23. The composition of claim 20 further comprising binders, lubricants, disintegrating agents, or coloring agents.

24. The composition of claim 23 wherein said binders are selected from the group consisting of starch, gelatin, natural sugars, corn sweeteners, natural and synthetic gums carboxymethylcellulose, polyethylene glycol, and waxes.

25. The composition of claim 23 wherein said lubricants are selected from the group consisting of sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, and sodium chloride.

26. The composition of claim 23 wherein said disintegrating agents are selected from the group consisting of starch, methyl cellulose, agar, bentonite, and xanthan gum.

27. The composition of claim 20 contained in a topical administration.

28. The composition of claim 27 wherein said active drug component can be admixed with carrier materials selected from the group consisting of alcohols, aloe vera gel, allontoin, glycerine, vitamin A and E oils, mineral oil, PPG2 myristyl propionate.

29. An oral composition comprising an EPO receptor modulating compound of claims 18 having an active drug component being combined with an oral, non-toxic pharmaceutically acceptable inert carrier.

30. The composition of claim 29 wherein said carrier is ethanol, glycerol, or water.

31. The composition of claim 29 further comprising binders, lubricants, disintegrating agents, or coloring agents.

32. The composition of claim 31 wherein said binders are selected from the group consisting of starch, gelatin, natural sugars, corn sweeteners, natural and synthetic gums carboxymethylcellulose, polyethylene glycol, and waxes.

33. The composition of claim 31 wherein said lubricants are selected from the group consisting of sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, and sodium chloride.

34. The composition of claim 31 wherein said disintegrating agents are selected from the group consisting of starch, methyl cellulose, agar, bentonite, and xanthan gum.

35. A compound of the formula:




wherein
Ra is H or t-bu;

when r=0 and n=1, then
Z is OH or t-Bu-O;
R1 and R4 are independently selected from the group consisting of nitro, phenoxy, phenylC1-5alkoxy, and substituted amino (where the substituents are selected from one or more members of the group consisting of C1-5alkyl, halosubstitutedC1-5alkyl, C1-5alknyl, C1-5alkenyl, phenyl, phenylC1-5alkyl, C1-5alkylcarbonyl, halosubstituted, C1-5alkylcarbonyl, carboxyC1-5alkyl, C1-5alkoxyC1-5alkyl, cinnamoyl, naphthylcarbonyl, furylcarbonyl, pyridylcarbonyl, C1-5alkylsulfonyl, phenylcarbonyl, phenylC1-5alkylcarbonyl, phenylsulfonyl, phenylC1-5alkylsulfonyl substituted phenylcarbonyl, substituted phenylC1-5alkylcarbonyl, substituted phenylsulfonyl, substituted phenylC1-5alkylsulfonyl, substituted phenyl, and substituted phenylC1-5alkyl[where the aromatic phenyl, phenylC1-5alkyl, phenylcarbonyl, phenylC1-5alkylcarbonyl, phenylsulfonyl, and phenylC1-5alkylsulfonyl substituents are independently selected from one to five members of the group consisting of C1-5alkyl, C1-5alkoxy, hydroxy, halogen, trifluoromethyl, nitro, cyano, and amino]); and
R3 and R5 are H;

when r=1 or 2 and n=2, then
Z is NH(CH2)3O(CH2)4O(CH2)3NH, NH(CH2)3O(CH2CH2O)2(CH2)3NH, or NH(CH2)2O(CH2)O(CH2)2NH
R2 is H or substituted amino (where the substituents are selected from one or more members of the group consisting of C1-5alkyl, halosubstitutedC1-5alkyl, C1-5alknyl, C1-5alkenyl, phenyl, phenylC1-5alkyl, C1-5alkylcarbonyl, halosubstituted, C1-5alkylcarbonyl, carboxyC1-5alkyl, C1-5alkoxyC1-5alkyl, cinnamoyl, naphthylcarbonyl, furylcarbonyl, pyridylcarbonyl, C1-5alkylsulfonyl, phenylcarbonyl, phenylC1-5alkylcarbonyl, phenylsulfonyl, phenylC1-5alkylsulfonyl substituted phenylcarbonyl, substituted phenylC1-5alkylcarbonyl, substituted phenylsulfonyl, substituted phenylC1-5alkylsulfonyl, substituted phenyl, and substituted phenylC1-5alkyl[where the aromatic phenyl, phenylC1-5alkyl, phenylcarbonyl, phenylC1-5alkylcarbonyl, phenylsulfonyl, and phenylC1-5alkylsulfonyl substituents are independently selected from one to five members of the group consisting of C1-5alkyl, C1-5alkoxy, hydroxy, halogen, trifluoromethyl, nitro, cyano, and amino]);
R4 is H or phenoxy; and
R3 and R5 are independently H or BnO.

36. A pharmaceutical composition comprising the compound of claim 35.

37. A pharmaceutical composition comprising an active drug component and the compound of claim 36.

38. The composition of claim 37 wherein said active drug component is combined with an oral, non-toxic pharmaceutically acceptable inert carrier.

39. The composition of claim 38 wherein said carrier is ethanol, glycerol, or water.

40. The composition of claim 37 further comprising binders, lubricants, disintegrating agents, or coloring agents.

41. The composition of claim 40 wherein said binders are selected from the group consisting of starch, gelatin, natural sugars, corn sweeteners, natural and synthetic gums carboxymethylcellulose, polyethylene glycol, and waxes.

42. The composition of claim 40 wherein said lubricants are selected from the group consisting of sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, and sodium chloride.

43. The composition of claim 40 wherein said disintegrating agents are selected from the group consisting of starch, methyl cellulose, agar, bentonite, and xanthan gum.

44. The composition of claim 37 contained in a topical administration.

45. The composition of claim 44 wherein said active drug component can be admixed with carrier materials selected from the group consisting of alcohols, aloe vera gel, allontoin, glycerine, vitamin A and E oils, mineral oil, PPG2 myristyl propionate.

46. An oral composition comprising an EPO receptor modulating compound of claim 35 having an active drug component being combined with an oral, non-toxic pharmaceutically acceptable inert carrier.

47. The composition of claim 46 wherein said carrier is ethanol, glycerol, or water.

48. The composition of claim 46 further comprising binders, lubricants, disintegrating agents, or coloring agents.

49. The composition of claim 48 wherein said binders are selected from the group consisting of starch, gelatin, natural sugars, corn sweeteners, natural and synthetic gums carboxymethylcellulose, polyethylene glycol, and waxes.

50. The composition of claim 48 wherein said lubricants are selected from the group consisting of sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, and sodium chloride.

51. The composition of claim 48 wherein said disintegrating agents are selected from the group consisting of starch, methyl cellulose, agar, bentonite, and xanthan gum.

52. A compound of the formula:




wherein
Ra and R9 are independently H or t-Bu;
R2 and R4 are independently H or phenoxy;
R3 and R5 are independently selected from the group consisting of H, BnO, and
substituted amino (where the substituents are selected from one or more members of the group consisting of C1-5alkyl, halosubstitutedC1-5alkyl, C1-5alknyl, C1-5alkenyl, phenyl, phenylC1-5alkyl, C1-5alkylcarbonyl, halosubstitutedC1-5alkylcarbonyl, carboxyC1-5alkyl, C1-5alkoxyC1-5alkyl, cinnamoyl, naphthylcarbonyl, furylcarbonyl, pyridylcarbonyl, C1-5alkylsulfonyl, phenylcarbonyl, phenylC1-5alkylcarbonyl, phenylsulfonyl, phenylC1-5alkylsulfonyl substituted phenylcarbonyl, substituted phenylC1-5alkylcarbonyl, substituted phenylsulfonyl, substituted phenylC1-5alkylsulfonyl, substituted phenyl, and substituted phenylC1-5alkyl, [where the aromatic phenyl, phenylC1-5alkyl, phenylcarbonyl, phenylC1-5alkylcarbonyl, phenylsulfonyl, and phenylC1-5alkylsulfonyl substituents are independently selected from one to five members of the group consisting of C1-5alkyl, C1-5alkoxy, hydroxy, halogen, trifluoromethyl, nitro, cyano, and amino]); and
n is 1 or 2.

53. A compound of the formula:




wherein
Ra is independently H or C1-5alkyl;
r is 1 or 2;
R2 and R4 are independently H, phenoxy, substituted phenoxy (where the substituents are selected from C1-5alkyl, C1-5alkoxy, hydroxy, halo, trifluoromethyl, nitro, cyano, and amino), 2,3-benzo, or 3,4 benzo;

when n is 1, then
Z is OH or C1-5alkoxy;
R3 and R5 are independently H or phenyl;

when n is 2, then
Z is NH(CH2)2O(CH2)2O(CH2)2NH, NH(CH2)3O(CH2)4O(CH2)3NH, NH(CH2)3O(CH2CH2O)2(CH2)3NH, NH(CH2)10NH, or NH(CH2)12NH; and
R3 and R5 are H.

54. A pharmaceutical composition comprising the compound of claim 53.

55. A pharmaceutical composition comprising an active drug component and the compound of claim 53.

56. The composition of claim 55 wherein said active drug component is combined with an oral, non-toxic pharmaceutically acceptable inert carrier.

57. The composition of claim 56 wherein said carrier is ethanol, glycerol, or water.

58. The composition of claim 55 further comprising binders, lubricants, disintegrating agents, or coloring agents.

59. The composition of claim 58 wherein said binders are selected from the group consisting of starch, gelatin, natural sugars, corn sweeteners, natural and synthetic gums carboxymethylcellulose, polyethylene glycol, and waxes.

60. The composition of claim 58 wherein said lubricants are selected from the group consisting of sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, and sodium chloride.

61. The composition of claim 58 wherein said disintegrating agents are selected from the group consisting of starch, methyl cellulose, agar, bentonite, and xanthan gum.

62. The composition of claim 59 contained in a topical administration.

63. The composition of claim 62 wherein said active drug component can be admixed with carrier materials selected from the group consisting of alcohols, aloe vera gel, allontoin, glycerine, vitamin A and E oils, mineral oil, PPG2 myristyl propionate.

64. An oral composition comprising an EPO receptor modulating compound of claim 53 having an active drug component being combined with an oral, non-toxic pharmaceutically acceptable inert carrier.

65. The composition of claim 64 wherein said carrier is ethanol, glycerol, or water.

66. The composition of claim 64 further comprising binders, lubricants, disintegrating agents, or coloring agents.

67. The composition of claim 66 wherein said binders are selected from the group consisting of starch, gelatin, natural sugars, corn sweeteners, natural and synthetic gums carboxymethylcellulose, polyethylene glycol, and waxes.

68. The composition of claim 66 wherein said lubricants are selected from the group consisting of sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, and sodium chloride.

69. The composition of claim 66 wherein said disintegrating agents are selected from the group consisting of starch, methyl cellulose, agar, bentonite, and xanthan gum.

70. A compound of the formula:




wherein:
Ra is H, Boc, or succinyl;

when n=1, then
R2 and R4 are independently selected from the group consisting of hydrogen, BnO, nitro, amino, substituted amino (where the substituents are selected from one or more members of the group consisting of C1-5alkyl, halosubstitutedC1-5alkyl, C1-5alknyl, C1-5alkenyl, phenyl, phenylC1-5alkyl, C1-5alkylcarbonyl, halosubstitutedC1-5alkylcarbonyl, carboxyC1-5alkyl, C1-5alkoxyC1-5alkyl, cinnamoyl, naphthylcarbonyl, furylcarbonyl, pyridylcarbonyl, C1-5alkylsulfonyl, phenylcarbonyl, phenylC1-5alkylcarbonyl, phenylsulfonyl, phenylC1-5alkylsulfonyl substituted phenylcarbonyl, substituted phenylC1-5alkylcarbonyl, substituted phenylsulfonyl, substituted phenylC1-5alkylsulfonyl, substituted phenyl, and substituted phenylC1-5alkyl, [where the aromatic phenyl, phenylC1-5alkyl, phenylcarbonyl, phenylC1-5alkylcarbonyl, phenylsulfonyl, and phenylC1-5alkylsulfonyl substituents are independently selected from one to five members of the group consisting of C1-5alkyl, C1-5alkoxy, hydroxy, halogen, trifluoromethyl, nitro, cyano, and amino]);
R3 and R5 are independently H or BnO; and
Z is alkoxy;

when n is 2 or 3, then
R2 and R4 are independently phenoxy or phenylC1-5alkoxy;
R3 and R5 are H; and
Z is NH(CH2)2O(CH2)2O(CH2)2NH, (NHCH2CH2)3N, or NH(CH2)3NMe(CH2)3NH.

71. A pharmaceutical composition comprising the compound of claim 70.

72. A pharmaceutical composition comprising an active drug component and the compound of claim 70.

73. The composition of claim 72 wherein said active drug component is combined with an oral, non-toxic pharmaceutically acceptable inert carrier.

74. The composition of claim 73 wherein said carrier is ethanol, glycerol, or water.

75. The composition of claim 72 further comprising binders, lubricants, disintegrating agents, or coloring agents.

76. The composition of claim 75 wherein said binders are selected from the group consisting of starch, gelatin, natural sugars, corn sweeteners, natural and synthetic gums carboxymethylcellulose, polyethylene glycol, and waxes.

77. The composition of claim 75 wherein said lubricants are selected from the group consisting of sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, and sodium chloride.

78. The composition of claim 75 wherein said disintegrating agents are selected from the group consisting of starch, methyl cellulose, agar, bentonite, and xanthan gum.

79. The composition of claim 72 contained in a topical administration.

80. The composition of claim 79 wherein said active drug component can be admixed with carrier materials selected from the group consisting of alcohols, aloe vera gel, allontoin, glycerine, vitamin A and E oils, mineral oil, PPG2 myristyl propionate.

81. An oral composition comprising an EPO receptor modulating compound of claim 70 having an active drug component being combined with an oral, non-toxic pharmaceutically acceptable inert carrier.

82. The composition of claim 81 wherein said carrier is ethanol, glycerol, or water.

83. The composition of claim 81 further comprising binders, lubricants, disintegrating agents, or coloring agents.

84. The composition of claim 83 wherein said binders are selected from the group consisting of starch, gelatin, natural sugars, corn sweeteners, natural and synthetic gums carboxymethylcellulose, polyethylene glycol, and waxes.

85. The composition of claim 83 wherein said lubricants are selected from the group consisting of sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, and sodium chloride.

86. The composition of claim 83 wherein said disintegrating agents are selected from the group consisting of starch, methyl cellulose, agar, bentonite, and xanthan gum.

87. A compound of the formula:




wherein:
Ra is H, Boc, or succinyl;
R2 is selected from the group consisting of phenoxy, substituted phenoxy (where the substituents are selected from C1-5alkyl, C1-5alkoxy, hydroxy, halo, trifluoromethyl, nitro, cyano, and amino), and
substituted amino (where the substituents are selected from one or more members of the group consisting of C1-5alkyl, halosubstitutedC1-5alkyl, C1-5alknyl, C1-5alkenyl, phenyl, phenylC1-5alkyl, C1-5alkylcarbonyl, halosubstitutedC1-5alkylcarbonyl, carboxyC1-5alkyl, C1-5alkoxyC1-5alkyl, cinnamoyl, naphthylcarbonyl, furylcarbonyl, pyridylcarbonyl, C1-5alkylsulfonyl, phenylcarbonyl, phenylC1-5alkylcarbonyl, phenylsulfonyl, phenylC1-5alkylsulfonyl substituted phenylcarbonyl, substituted phenylC1-5alkylcarbonyl, substituted phenylsulfonyl, substituted phenylC1-5alkylsulfonyl, substituted phenyl, and substituted phenylC1-5alkyl, [where the aromatic phenyl, phenylC1-5alkyl, phenylcarbonyl, phenylC1-5alkylcarbonyl, phenylsulfonyl, and phenylC1-5alkylsulfonyl substituents are independently selected from one to five members of the group consisting of C1-5alkyl, C1-5alkoxy, hydroxy, halogen, trifluoromethyl, nitro, cyano, and amino]);
R4 and R5 are independently H, methoxy, phenoxy, BnO, halo, or 2,3-benzo; and
R9 is H or methyl.

88. A pharmaceutical composition comprising the compound of claim 87.

89. A pharmaceutical composition comprising an active drug component and the compound of claim 87.

90. The composition of claim 89 wherein said active drug component is combined with an oral, nontoxic pharmaceutically acceptable inert carrier.

91. The composition of claim 90 wherein said carrier is ethanol, glycerol, or water.

92. The composition of claim 89 further comprising binders, lubricants, disintegrating agents, or coloring agents.

93. The composition of claim 92 wherein said binders are selected from the group consisting of starch, gelatin, natural sugars, corn sweeteners, natural and synthetic gums carboxymethylcellulose, polyethylene glycol, and waxes.

94. The composition of claim 92 wherein said lubricants are selected from the group consisting of sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, and sodium chloride.

95. The composition of claim 92 wherein said disintegrating agents are selected from the group consisting of starch, methyl cellulose, agar, bentonite, and xanthan gum.

96. The composition of claim 89 contained in a topical administration.

97. The composition of claim 96 wherein said active drug component can be admixed with carrier materials selected from the group consisting of alcohols, aloe vera gel, allontoin, glycerine, vitamin A and E oils, mineral oil, PPG2 myristyl propionate.

98. An oral composition comprising an EPO receptor modulating compound of claim 87 having an active drug component being combined with an oral, non-toxic pharmaceutically acceptable inert carrier.

99. The composition of claim 98 wherein said carrier is ethanol, glycerol, or water.

100. The composition of claim 98 further comprising binders, lubricants, disintegrating agents, or coloring agents.

101. The composition of claim 100 wherein said binders are selected from the group consisting of starch, gelatin, natural sugars, corn sweeteners, natural and synthetic gums carboxymethylcellulose, polyethylene glycol, and waxes.

102. The composition of claim 100 wherein said lubricants are selected from the group consisting of sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, and sodium chloride.

103. The composition of claim 100 wherein said disintegrating agents are selected from the group consisting of starch, methyl cellulose, agar, bentonite, and xanthan gum.

104. A compound of the formula:




wherein:
Ra is H or the side chain of a natural or unnatural α-amino acid, where if said side chain contains a protectable group, that group may be protected with a member of the group consisting of succinyl, glutaryl, 3,3-dimethylglutaryl, C1-5alkyl, C1-5alkoxycarbonyl, acetyl, N-(9-fluorenylmethoxycarbonyl), trifluoroacetyl, omega-carboxyC1-5alkylcarbonyl, t-butoxycarbonyl, benzyl, benzyloxycarbonyl, 2-chlorobenzyloxycarbonyl, phenylsulfonyl, ureido, t-butyl, cinnamoyl, trityl, 4-methyltrityl, 1-(4,4-dimethyl-2,6-dioxocyclohexylidene)ethyl, tosyl, 4-methoxy-2,3,6-trimethylbenzenesulfonyl, phenylureido, and substituted phenylureido(where the phenyl substituents are phenoxy, halo, C1-5alkoxycarbonyl);
R2 and R4 are independently selected from the group consisting of H, nitro, amino, phenoxy,
substituted phenoxy (where the substituents are selected from C1-5alkyl, C1-5alkoxy, hydroxy, halo, trifluoromethyl, nitro, cyano, and amino),
substituted amino (where the substituents are selected from one or more members of the group consisting of C1-5alkyl, halosubstitutedC1-5alkyl, C1-5alknyl, C1-5alkenyl, phenyl, phenylC1-5alkyl, C1-5alkylcarbonyl, halosubstitutedC1-5alkylcarbonyl, carboxyC1-5alkyl, C1-5alkoxyC1-5alkyl, cinnamoyl, naphthylcarbonyl, furylcarbonyl, pyridylcarbonyl, C1-5alkylsulfonyl, phenylcarbonyl, phenylC1-5alkylcarbonyl, phenylsulfonyl, phenylC1-5alkylsulfonyl substituted phenylcarbonyl, substituted phenylC1-5alkylcarbonyl, substituted phenylsulfonyl, substituted phenylC1-5alkylsulfonyl, substituted phenyl, and substituted phenylC1-5alkyl, [where the aromatic phenyl, phenylC1-5alkyl, phenylcarbonyl, phenylC1-5alkylcarbonyl, phenylsulfonyl, and phenylC1-5alkylsulfonyl substituents are independently selected from one to five members of the group consisting of C1-5alkyl, C1-5alkoxy, hydroxy, halogen, trifluoromethyl, nitro, cyano, and amino]);
R3 and R5 are independently H, phenyl, C1-5alkoxy, 3,4 benzo, or nitro; and
Z is H, C1-5alkoxy, substituted phenyl (where the substituents are selected from C1-5alkyl, C1-5alkoxy, hydroxy, halo, trifluoromethyl, nitro, cyano, and amino), or
substituted amino (where the substituents are selected from one or more members of the group consisting of C1-5alkyl, halosubstitutedC1-5alkyl, C1-5alknyl, C1-5alkenyl, phenyl, phenylC1-5alkyl, C1-5alkylcarbonyl, halosubstitutedC1-5alkylcarbonyl, carboxyC1-5alkyl, C1-5alkoxyC1-5alkyl, cinnamoyl, naphthylcarbonyl, furylcarbonyl, pyridylcarbonyl, C1-5alkylsulfonyl, phenylcarbonyl, phenylC1-5alkylcarbonyl, phenylsulfonyl, phenylC1-5alkylsulfonyl substituted phenylcarbonyl, substituted phenylC1-5alkylcarbonyl, substituted phenylsulfonyl, substituted phenylC1-5alkylsulfonyl, substituted phenyl, and substituted phenylC1-5alkyl, [where the aromatic phenyl, phenylC1-5alkyl, phenylcarbonyl, phenylC1-5alkylcarbonyl, phenylsulfonyl, and phenylC1-5alkylsulfonyl substituents are independently selected from one to five members of the group consisting of C1-5alkyl, C1-5alkoxy, hydroxy, halogen, trifluoromethyl, nitro, cyano, and amino]).

105. A pharmaceutical composition comprising the compound of claim 104.

106. A pharmaceutical composition comprising an active drug component and the compound of claim 104.

107. The composition of claim 106 wherein said active drug component is combined with an oral, non-toxic pharmaceutically acceptable inert carrier.

108. The composition of claim 107 wherein said carrier is ethanol, glycerol, or water.

109. The composition of claim 106 further comprising binders, lubricants, disintegrating agents, or coloring agents.

110. The composition of claim 109 wherein said binders are selected from the group consisting of starch, gelatin, natural sugars, corn sweeteners, natural and synthetic gums carboxymethylcellulose, polyethylene glycol, and waxes.

111. The composition of claim 109 wherein said lubricants are selected from the group consisting of sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, and sodium chloride.

112. The composition of claim 109 wherein said disintegrating agents are selected from the group consisting of starch, methyl cellulose, agar, bentonite, and xanthan gum.

113. The composition of claim 106 contained in a topical administration.

114. The composition of claim 113 wherein said active drug component can be admixed with carrier materials selected from the group consisting of alcohols, aloe vera gel, allontoin, glycerine, vitamin A and E oils, mineral oil, PPG2 myristyl propionate.

115. An oral composition comprising an EPO receptor modulating compound of claim 104 having an active drug component being combined with an oral, non-toxic pharmaceutically acceptable inert carrier.

116. The composition of claim 115 wherein said carrier is ethanol, glycerol, or water.

117. The composition of claim 115 further comprising binders, lubricants, disintegrating agents, or coloring agents.

118. The composition of claim 117 wherein said binders are selected from the group consisting of starch, gelatin, natural sugars, corn sweeteners, natural and synthetic gums carboxymethylcellulose, polyethylene glycol, and waxes.

119. The composition of claim 117 wherein said lubricants are selected from the group consisting of sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, and sodium chloride.

120. The composition of claim 117 wherein said disintegrating agents are selected from the group consisting of starch, methyl cellulose, agar, bentonite, and xanthan gum.

121. A compound of the formula:




wherein:
Ra is H, boc, succinyl, glutaryl, or 3,3-dimethylglutaryl; and
Z is NH(CH2)3O(CH2CH2O)2(CH2)3NH, NH(CH2)3O(CH2)4O(CH2)3NH, NH(CH2)12NH, or NH(CH2)2O(CH2)2O(CH2)2NH.

122. A pharmaceutical composition comprising the compound of claim 121.

123. A pharmaceutical composition comprising an active drug component and the compound of claims 121.

124. The composition of claim 123 wherein said active drug component is combined with an oral, non-toxic pharmaceutically acceptable inert carrier.

125. The composition of claim 124 wherein said carrier is ethanol, glycerol, or water.

126. The composition of claim 123 further comprising binders, lubricants, disintegrating agents, or coloring agents.

127. The composition of claim 126 wherein said binders are selected from the group consisting of starch, gelatin, natural sugars, corn sweeteners, natural and synthetic gums carboxymethylcellulose, polyethylene glycol, and waxes.

128. The composition of claim 126 wherein said lubricants are selected from the group consisting of sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, and sodium chloride.

129. The composition of claim 126 wherein said disintegrating agents are selected from the group consisting of starch, methyl cellulose, agar, bentonite, and xanthan gum.

130. The composition of claim 123 contained in a topical administration.

131. The composition of claim 130 wherein said active drug component can be admixed with carrier materials selected from the group consisting of alcohols, aloe vera gel, allontoin, glycerine, vitamin A and E oils, mineral oil, PPG2 myristyl propionate.

132. An oral composition comprising an EPO receptor modulating compound of claim 121 having an active drug component being combined with an oral, non-toxic pharmaceutically acceptable inert carrier.

133. The composition of claim 132 wherein said carrier is ethanol, glycerol, or water.

134. The composition of claim 133 further comprising binders, lubricants, disintegrating agents, or coloring agents.

135. The composition of claim 134 wherein said binders are selected from the group consisting of starch, gelatin, natural sugars, corn sweeteners, natural and synthetic gums carboxymethylcellulose, polyethylene glycol, and waxes.

136. The composition of claim 134 wherein said lubricants are selected from the group consisting of sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, and sodium chloride.

137. The composition of claim 134 wherein said disintegrating agents are selected from the group consisting of starch, methyl cellulose, agar, bentonite, and xanthan gum.

138. A compound of the formula:




wherein:
R1 is H or C1-5 alkyl;
R2 is H, BnO, or substituted phenoxy (where the substituents are selected from C1-5alkyl, C1-5alkoxy, hydroxy, halo, trifluoromethyl, nitro, cyano, and amino); and
R3 is H or phenoxy.

139. A pharmaceutical composition comprising the compound of claim 138.

140. A pharmaceutical composition comprising an active drug component and the compound of claim 138.

141. The composition of claim 140 wherein said active drug component is combined with an oral, non-toxic pharmaceutically acceptable inert carrier.

142. The composition of claim 141 wherein said carrier is ethanol, glycerol, or water.

143. The composition of claim 140 further comprising binders, lubricants, disintegrating agents, or coloring agents.

144. The composition of claim 143 wherein said binders are selected from the group consisting of starch, gelatin, natural sugars, corn sweeteners, natural and synthetic gums carboxymethylcellulose, polyethylene glycol, and waxes.

145. The composition of claim 143 wherein said lubricants are selected from the group consisting of sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, and sodium chloride.

146. The composition of claim 143 wherein said disintegrating agents are selected from the group consisting of starch, methyl cellulose, agar, bentonite, and xanthan gum.

147. The composition of claim 140 contained in a topical administration.

148. The composition of claim 147 wherein said active drug component can be admixed with carrier materials selected from the group consisting of alcohols, aloe vera gel, allontoin, glycerine, vitamin A and E oils, mineral oil, PPG2 myristyl propionate.

149. An oral composition comprising an EPO receptor modulating compound of claim 138 having an active drug component being combined with an oral, non-toxic pharmaceutically acceptable inert carrier.

150. The composition of claim 149 wherein said carrier is ethanol, glycerol, or water.

151. The composition of claim 149 further comprising binders, lubricants, disintegrating agents, or coloring agents.

152. The composition of claim 151 wherein said binders are selected from the group consisting of starch, gelatin, natural sugars, corn sweeteners, natural and synthetic gums carboxymethylcellulose, polyethylene glycol, and waxes.

153. The composition of claim 151 wherein said lubricants are selected from the group consisting of sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, and sodium chloride.

154. The composition of claim 151 wherein said disintegrating agents are selected from the group consisting of starch, methyl cellulose, agar, bentonite, and xanthan gum.

155. A compound of the formula:




wherein:
R1 is H or methyl;
R2 is H, phenoxy, or substituted phenyl (where the substituents are selected from C1-5alkyl, C1-5alkoxy, hydroxy, halo, trifluoromethyl, nitro, cyano, and amino);
R3 is H or phenyl; and
R4 is H or t-Bu.

156. A pharmaceutical composition comprising the compound of claim 155.

157. A pharmaceutical composition comprising an active drug component and a compound of claim 155.

158. The composition of claim 157 wherein said active drug component is combined with an oral, non-toxic pharmaceutically acceptable inert carrier.

159. The composition of claim 158 wherein said carrier is ethanol, glycerol, or water.

160. The composition of claim 157 further comprising binders, lubricants, disintegrating agents, or coloring agents.

161. The composition of claim 160 wherein said binders are selected from the group consisting of starch, gelatin, natural sugars, corn sweeteners, natural and synthetic gums carboxymethylcellulose, polyethylene glycol, and waxes.

162. The composition of claim 160 wherein said lubricants are selected from the group consisting of sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, and sodium chloride.

163. The composition of claim 160 wherein said disintegrating agents are selected from the group consisting of starch, methyl cellulose, agar, bentonite, and xanthan gum.

164. The composition of claim 157 contained in a topical administration.

165. The composition of claim 164 wherein said active drug component can be admixed with carrier materials selected from the group consisting of alcohols, aloe vera gel, allontoin, glycerine, vitamin A and E oils, mineral oil, PPG2 myristyl propionate.

166. An oral composition comprising an EPO receptor modulating compound of claim 155 having an active drug component being combined with an oral, non-toxic pharmaceutically acceptable inert carrier.

167. The composition of claim 166 wherein said carrier is ethanol, glycerol, or water.

168. The composition of claim 166 further comprising binders, lubricants, disintegrating agents, or coloring agents.

169. The composition of claim 168 wherein said binders are selected from the group consisting of starch, gelatin, natural sugars, corn sweeteners, natural and synthetic gums carboxymethylcellulose, polyethylene glycol, and waxes.

170. The composition of claim 168 wherein said lubricants are selected from the group consisting of sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, and sodium chloride.

171. The composition of claim 168 wherein said disintegrating agents are selected from the group consisting of starch, methyl cellulose, agar, bentonite, and xanthan gum.

172. A compound of the formula:




wherein:
R1 is H or t-Bu;
R2 is phenoxy; and
R3 is H.

173. A pharmaceutical composition comprising the compound of claim 172.

174. A pharmaceutical composition comprising an active drug component and the compound of claim 172.

175. The composition of claim 174 wherein said active drug component is combined with an oral, non-toxic pharmaceutically acceptable inert carrier.

176. The composition of claim 175 wherein said carrier is ethanol, glycerol, or water.

177. The composition of claim 174 further comprising binders, lubricants, disintegrating agents, or coloring agents.

178. The composition of claim 177 wherein said binders are selected from the group consisting of starch, gelatin, natural sugars, corn sweeteners, natural and synthetic gums carboxymethylcellulose, polyethylene glycol, and waxes.

179. The composition of claim 177 wherein said lubricants are selected from the group consisting of sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, and sodium chloride.

180. The composition of claim 177 wherein said disintegrating agents are selected from the group consisting of starch, methyl cellulose, agar, bentonite, and xanthan gum.

181. The composition of claim 174 contained in a topical administration.

182. The composition of claim 181 wherein said active drug component can be admixed with carrier materials selected from the group consisting of alcohols, aloe vera gel, allontoin, glycerine, vitamin A and E oils, mineral oil, PPG2 myristyl propionate.

183. An oral composition comprising an EPO receptor modulating compound as in one of claim 174 having an active drug component being combined with an oral, non-toxic pharmaceutically acceptable inert carrier.

184. The composition of claim 183 wherein said carrier is ethanol, glycerol, or water.

185. The composition of claim 183 further comprising binders, lubricants, disintegrating agents, or coloring agents.

186. The composition of claim 183 wherein said binders are selected from the group consisting of starch, gelatin, natural sugars, corn sweeteners, natural and synthetic gums carboxymethylcellulose, polyethylene glycol, and waxes.

187. The composition of claim 185 wherein said lubricants are selected from the group consisting of sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, and sodium chloride.

188. The composition of claim 185 wherein said disintegrating agents are selected from the group consisting of starch, methyl cellulose, agar, bentonite, and xanthan gum.

189. A compound of the formula:



wherein:
R1 is H or methyl; R2 is H, phenoxy, substituted phenyl (where the substituents are selected from C1-5alkyl, C1-5alkoxy, hydroxy, halo, trifluoromethyl, nitro, cyano, and amino), or CO2Me;
R3 is H, trifluoromethyl, nitro, C1-5alkoxy, carboxyl, or 2,3-benzo; and R4 is substituted amino (where the substituents are selected from one or more members of the group consisting of C1-5alkyl, halosubstituted C1-5alkyl, C1-5alkynl, C1-5alkenyl, phenyl, phenyl C1-5alkyl, C1-5alkylcarbonyl, halosubstituted C1-5alk-ylcarbonyl, carboxy C1-5alkyl, C1-5alkoxy C1-5alkyl, cinnamoyl, naphthylcarbonyl, furylcarbonyl, pyridylcarbonyl, C1-5alkylsulfonyl, phenylcarbonyl, phenyl C1-5alkylcarbonyl, phenylsulfonyl, phenyl C1-5alkylsulfonyl substituted phenylcarbonyl, substituted phenyl C1-5alkylcarbonyl, substituted phenylsulfonyl, substituted phenyl C1-5alkylsulfonyl, substituted phenyl, and substitued phenyl C1-5alkyl, [where the aromatic phenyl, phenyl C1-5alkyl, phenylcarbonyl, phenyl C1-5alkylcarbonyl, phenylsulfonyl, and phenyl C1-5alkylsulfonyl substituents are independently selected from one to five members of the group consisting of C1-5alkyl, C1-5alkoxy, hydroxy, halogen, trifluoromethyl, nitro, cyano, and amino]).

190. A pharmaceutical composition comprising the compound of claim 189.

191. A pharmaceutical composition comprising an active drug component and the compound of claim 189.

192. The composition of claim 191 wherein said active drug component is combined with an oral, non-toxic pharmaceutically acceptable inert carrier.

193. The composition of claim 192 wherein said carrier is ethanol, glycerol, or water.

194. The composition of claim 191 further comprising binders, lubricants, disintegrating agents, or coloring agents.

195. The composition of claim 194 wherein said binders are selected from the group consisting of starch, gelatin, natural sugars, corn sweeteners, natural and synthetic gums carboxymethylcellulose, polyethylene glycol, and waxes.

196. The composition of claim 194 wherein said lubricants are selected from the group consisting of sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, and sodium chloride.

197. The composition of claim 194 wherein said disintegrating agents ors are selected from the group consisting of starch, methyl cellulose, agar, bentonite, and xanthan gum.

198. The composition of claim 191 contained in a topical administration.

199. The composition of claim 198 wherein said active drug component can be admixed with carrier materials selected from the group consisting of alcohols, aloe vera gel, allontoin, glycerine, vitamin A and E oils, mineral oil, PPG2 myristyl propionate.

200. An oral composition comprising an EPO receptor modulating compound of claim 189 having an active drug component being combined with an oral, non-toxic pharmaceutically acceptable inert carrier.

201. The composition of claim 200 wherein said carrier is ethanol, glycerol, or water.

202. The composition of claim 200 further comprising binders, lubricants, disintegrating agents, or coloring agents.

203. The composition of claim 202 wherein said binders are selected from the group consisting of starch, gelatin, natural sugars, corn sweeteners, natural and synthetic gums carboxymethycellulose, polyethylene glycol, and waxes.

204. The composition of claim 202 wherein said lubricants are selected from the group consisting of sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, and sodium chloride.

205. The composition of claim 202 wherein said disintegrating agents ors are selected from the group consisting of starch, methyl cellulose, agar, bentonite, and xanthan gum.