WO1997012903A1 - Compounds, compositions and methods for inhibiting the binding of proteins containing an sh2 domain to cognate phosphorylated proteins - Google Patents

Compounds, compositions and methods for inhibiting the binding of proteins containing an sh2 domain to cognate phosphorylated proteins Download PDF

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Publication number
WO1997012903A1
WO1997012903A1 PCT/US1996/015998 US9615998W WO9712903A1 WO 1997012903 A1 WO1997012903 A1 WO 1997012903A1 US 9615998 W US9615998 W US 9615998W WO 9712903 A1 WO9712903 A1 WO 9712903A1
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WIPO (PCT)
Prior art keywords
phenyl
acetylamino
carbamoyl
methyl
alkyl
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PCT/US1996/015998
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French (fr)
Inventor
Elizabeth A. Lunney
Kimberly Suzanne Para
Mark Stephen Plummer
Josyula Venkata Nagendra Vara Prasad
Alan Robert Saltiel
Tomi Sawyer
Aurash Shahripour
Juswinder Singh
Charles John Stankovic
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Warner-Lambert Company
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Priority to US09/051,038 priority Critical patent/US5922697A/en
Priority to AU73926/96A priority patent/AU7392696A/en
Publication of WO1997012903A1 publication Critical patent/WO1997012903A1/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
    • C07F9/02—Phosphorus compounds
    • C07F9/06—Phosphorus compounds without P—C bonds
    • C07F9/08—Esters of oxyacids of phosphorus
    • C07F9/09—Esters of phosphoric acids
    • C07F9/12—Esters of phosphoric acids with hydroxyaryl compounds
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
    • C07F9/02—Phosphorus compounds
    • C07F9/28—Phosphorus compounds with one or more P—C bonds
    • C07F9/38—Phosphonic acids [RP(=O)(OH)2]; Thiophosphonic acids ; [RP(=X1)(X2H)2(X1, X2 are each independently O, S or Se)]
    • C07F9/3804—Phosphonic acids [RP(=O)(OH)2]; Thiophosphonic acids ; [RP(=X1)(X2H)2(X1, X2 are each independently O, S or Se)] not used, see subgroups
    • C07F9/3834—Aromatic acids (P-C aromatic linkage)
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
    • C07F9/02—Phosphorus compounds
    • C07F9/28—Phosphorus compounds with one or more P—C bonds
    • C07F9/38—Phosphonic acids [RP(=O)(OH)2]; Thiophosphonic acids ; [RP(=X1)(X2H)2(X1, X2 are each independently O, S or Se)]
    • C07F9/3804—Phosphonic acids [RP(=O)(OH)2]; Thiophosphonic acids ; [RP(=X1)(X2H)2(X1, X2 are each independently O, S or Se)] not used, see subgroups
    • C07F9/3882—Arylalkanephosphonic acids
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
    • C07F9/02—Phosphorus compounds
    • C07F9/28—Phosphorus compounds with one or more P—C bonds
    • C07F9/38—Phosphonic acids [RP(=O)(OH)2]; Thiophosphonic acids ; [RP(=X1)(X2H)2(X1, X2 are each independently O, S or Se)]
    • C07F9/40—Esters thereof
    • C07F9/4003—Esters thereof the acid moiety containing a substituent or a structure which is considered as characteristic
    • C07F9/4056—Esters of arylalkanephosphonic acids
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
    • C07F9/02—Phosphorus compounds
    • C07F9/28—Phosphorus compounds with one or more P—C bonds
    • C07F9/38—Phosphonic acids [RP(=O)(OH)2]; Thiophosphonic acids ; [RP(=X1)(X2H)2(X1, X2 are each independently O, S or Se)]
    • C07F9/40—Esters thereof
    • C07F9/4071—Esters thereof the ester moiety containing a substituent or a structure which is considered as characteristic
    • C07F9/4075—Esters with hydroxyalkyl compounds
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06191—Dipeptides containing heteroatoms different from O, S, or N
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • This invention relates to compounds that inhibit the binding of proteins containing an SH2 domain to cognate phosphorylated proteins. This invention also relates to pharmaceutical compositions containing the compounds and to therapeutic methods that use the compounds.
  • tyrosine kinases Many of the signal transduction pathways that regulate a variety of cellular processes, including the differentiation and proliferation of normal and malignant cells, operate through phosphorylated proteins called tyrosine kinases.
  • the two major types of tyrosine kinases are receptor and nonreceptor tyrosine kinases.
  • Receptor tyrosine kinases contain binding sites or receptors for growth factors such as epidermal growth factor (EGF) or platelet-derived growth factor (PDGF) .
  • EGF epidermal growth factor
  • PDGF platelet-derived growth factor
  • the tyrosine kinase receptor protein When a growth factor such as EGF or PDGF binds to the receptor, the tyrosine kinase receptor protein is activated; the tyrosine kinase receptor protein is autophosphorylated; and endogenous proteins that participate in the signal transduction pathway are phosphorlyated.
  • SH2 domain a specific domain that provides for interaction with an activated tyrosine kinase receptor protein. This interaction is a protein- protein interaction.
  • One protein that contains an SH2 domain is pp60c-src kinase, which is also a tyrosine kinase.
  • the pp60c-src kinase is a nonreceptor kinase and is related to a number of nonreceptor tyrosine kinases, which include, but are not limited to, Fyn, Lck, Yes, Blk, Lyn, Fgr, Hck, and Yrk.
  • the pp60c-src protein is exemplary of the Src family of tyrosine kinases.
  • the pp60c-src has three major domains: SHI, SH2, and SH3.
  • SHI domain is most commonly called the catalytic domain or tyrosine kinase domain.
  • SH3 domain is a binding region for proteins having proline-rich sequences. Both the SH2 and SH3 domains are noncatalytic, but are important in protein-protein recognition.
  • the Src family of protein kinases which all contain a SH2 domain, are involved in a number of cellular signalling pathways. For example, Src is involved in growth factor receptor signalling; integrin-mediated signaling; T- and B-cell activation and osteoclast activation. It is known that the Src SH2 domain binds to several key receptor and nonreceptor tyrosine kinases such as tyrosine kinases containing receptors for PDGF, EGF, HER2/Neu (an oncogene form of EGF), Fibroblast growth factor, focal adhesion kinase, pl30 protein, and p68 protein. In addition, pp60c-src has been shown to be involved in the regulation of DNA synthesis, mitosis, and other cellular activities.
  • U is -CO-, -CS-, -SO-, or -S0 2 ' Y is
  • X is or a bond
  • R 1 is hydrogen, RCONR'-, RR'NCONR''-, RS0 2 NR'-, RCSNR'-, RR'NCSNR''-, RR'NS0 2 NR' ' - , ROCONR'-, or . .
  • R 2 is hydrogen, alkyl, cycloalkyl- (CH 2 ) n - , substituted alkyl, aryl- (CH 2 ) n - , heteroaryl- (CH 2 ) n - , - (CH 2 ) n -C0 2 H, substituted cycloalkyl- (CH 2 ) n - , substituted aryl- (CH 2 ) n - or substituted heteroaryl- (CH 2 ) n _ ;
  • R 3 is hydrogen, alkyl, cycloalkyl- (CH 2 ) n - , substituted alkyl, aryl- (CH 2 ) n - , heteroaryl- (CH 2 ) n - , -(CH 2 ) n -C0 2 H, substituted cycloalkyl- (CH 2 ) n -, substituted aryl- (CH 2 ) n - , or substituted heteroaryl- (CH 2 ) n - ;
  • R 4 is hydrogen or alkyl
  • R 6 and R 9 are independently -OP0 3 R c R d , -CF 2 P0 3 R c R d , -CH 2 P0 3 R c R d , -P0 3 R c R d , -S0 3 R c , -OS0 3 R c , -CH 2 S0 3 R c , -S0 2 NH 2 , -OS0 2 NH 2 , or -CH 2 S0 2 NH 2 ;
  • R 7' and Rft° are independently hydrogen, alkyl, substituted alkyl, halogen, -OR, -NRRX -COCF 3 , -(CH 2 ) n CH 2 OH, -(CH 2 ) n C0 2 H, -(CH 2 ) n CHO, -(CH 2 ) n NRRX or -Q-CH 2 -(CH 2 ) n -NRR';
  • R 10 is -(CH 2 ) n C0 2 H, hydrogen, alkyl, aryl, substituted alkyl, or - (CH 2 ) n -substituted aryl;
  • R c and R d are independently -R, -CH 2 CH 2 Z,
  • Q is -0-, -NH-, -S-, -CH 2 0-, -CH 2 NH-, or -CH 2 S-;
  • Z is -Cl, -Br, or -F;
  • R, R', and R' ' are independently hydrogen, alkyl, cycloalkyl-(CH 2 ) n -, aryl-(CH 2 ) n -, heteroaryl-(CH 2 ) n ", substituted alkyl, substituted cycloalkyl-(CH 2 ) n _ , substituted aryl- (CH 2 ) n _ , -(CH2) n C0 H, or substituted heteroaryl-(CH 2 ) n -; and each n is independently 0 to 5, or the pharmaceutically acceptable salts, amides, esters, or prodrugs thereof.
  • X is R- ⁇ c
  • R 1 is RCONR'-, -NRCONR'R'X -NRS0 2 RX or
  • R 2 , R 4 , R 7 , and R 8 are hydrogen
  • R 3 is -(CH 2 ) n C0 2 H, alkyl, or - (CH 2 ) n -substituted aryl;
  • R' is -NRR', -NCH(CH )R, or
  • R 6 is -0P0 3 R c R d , -CF 2 P0 3 R c R d , or -P0 3 R c R d .
  • R 6 is -0P0 3 R c R d , -CF 2 P0 3 R c R d , or -P0 3 R c R d .
  • X is R 1 R 2 C and ⁇
  • R 8 are hydrogen; R 3 is -CH 2 CH 2 C0 2 H;
  • R 5 is -NRR', - ;
  • R R 6 is -OP0 3 R c R d , -CF 2 P0 3 R c R d , or -P0 3 R c R d .
  • R R 6 is -OP0 3 R c R d , -CF 2 P0 3 R c R d , or -P0 3 R c R d .
  • R 3 and R 10 are -(CH 2 ) n C0 2 H; R 4 is hydrogen;
  • R 7 and R 8 are hydrogen; and R 6 is -OP0 3 R c R d , -CF 2 P0 3 R c R d , or -P0 3 R c R d .
  • the present invention provides the compounds of Formula II below:
  • U and W are independently -CO-, -CS-, -SO-, or -S0 2 -;
  • R 11 is hydrogen, alkyl, -OH, substituted alkyl, or
  • R- 1*- is hydrogen or alkyl
  • R 13 is -(CH 2 ) n C0 2 H, alkyl, - (CH 2 ) n -aryl, - (CH 2 ) n -heteroaryl, - (CH 2 ) n -cycloalkyl, hydrogen, substituted cycloalkyl- (CH 2 ) n - , substituted aryl- (CH 2 ) n - , substituted heteroaryl- (CH 2 ) n - , or substituted alkyl;
  • R 14 and R 17 are independently -OP0 3 R c R d , -CF 2 P0 3 R c R d , -CH 2 P0 3 R c R d , -P0 3 R c R d , -S0 3 R c , -OS0 3 R c , -CH 2 S0 3 R c , -S0 2 NH 2 , -OS0 2 NH 2 , or -CH 2 S0 2 NH 2 ;
  • R 15 and R 16 are independently hydrogen, alkyl, halogen, -OR, -NRRX -COCF3, - (CH 2 ) n CH 2 OH, -(CH 2 ) n C0 2 H, -(CH 2 ) n NRRX -(CH 2 ) n CHO, or -Q-CH 2 " (CH 2 ) n -NRR' ;
  • R 18 is -(CH 2 ) n C0 2 R, hydrogen, alkyl, - (CH 2 ) n CONRR' , substituted alkyl, or - (CH 2 ) n -substituted aryl;
  • R 19 is hydrogen, RCONR'-, RR'NCONR''-, RS0 2 NR'
  • R 20 is hydrogen, alkyl, cycloalkyl- (CH 2 ) n - / substituted alkyl, aryl- (CH 2 ) n " , heteroaryl- (CH 2 ) n -, - (CH 2 ) n -C0 2 H, substituted cycloalkyl- (CH 2 ) n - , substituted aryl- (CH 2 ) n ⁇ , or substituted heteroaryl- (CH 2 ) n ⁇ ;
  • R a is hydrogen, halogen, or alkyl;
  • R b is hydrogen, alkyl, -OR, -0(CH 2 ) n -aryl, -NRRX -0(CH 2 ) n -substituted alkyl, -SR, -0(CH 2 ) n -substituted aryl, or -0(CH 2 ) n -cycloalkyl;
  • R c and R d are independently -R, -CH2CH2Z, —o 0 R ,
  • Q is -0-, -NH-, -S-, -CH 2 0-, -CH 2 NH-, or -CH 2 S-;
  • Z is -Cl, -Br, or -F;
  • R, RX and R' ' are independently hydrogen, alkyl, cycloalkyl- (CH 2 ) n - , aryl- (CH 2 ) n - , heteroaryl- (CH 2 ) n - , -CH 2 -(CH 2 ) n -C0 2 H, substituted cycloalkyl- (CH 2 ) n - , substituted alkyl, substituted aryl- (CH ) n - , or substituted heteroaryl- (CH 2 ) n - ; and each n is independently 0 to 5, or the pharmaceutically acceptable salts, amides, esters, or prodrugs thereof.
  • R ,1 ⁇ 3 J , R ⁇ , , and R 1 ⁇ 6 ⁇ are hydrogen
  • R and R 4 -- 1 - are hydrogen or alkyl
  • R b is -OR, -0(CH 2 ) n -aryl, -0(CH 2 ) n substituted aryl, or -0(CH 2 ) n cycloalkyl;
  • X is R 19 R 0 c ⁇
  • U and W are -CO- .
  • X is R 19 R 20 C ⁇
  • R 19 is RCONR ' - or RR ' NCONR ' ' - ;
  • R 20 , R 15 , R 13 , R 11 , R a , and R 16 are hydrogen;
  • R 12 is alkyl or hydrogen;
  • R b is -OR, -0(CH 2 ) n -aryl, -0(CH 2 ) n substituted aryl, or -0(CH 2 ) n cycloalkyl;
  • R 14 is -OP0 3 R c R d or -CF 2 P0 3 R c R d .
  • Also provided by the present invention is method of inhibiting the binding of a protein containing an SH2 domain to a cognate phosphorylated protein, the method comprising administering to a patient in need of SH2 inhibition an SH2 inhibiting amount of a compound of Formula I or II.
  • the present invention provides a pharmaceutical composition that comprises a compound of Formula I or II and a pharmaceutically acceptable carrier.
  • the present invention provides a method of treating a patient having a proliferative disease, the method comprising administering to the patient a therapeutically effective amount of a compound of Formula I or II.
  • the present invention provides a method of treating a patient having cancer, the method comprising administering to the patient a therapeutically effective oxygen of Formula I or II.
  • the present invention provides a method of treating a patient having restenosis, the method comprising administering to the patient a therapeutically effective amount of a compound of Formula I or II.
  • the present invention provides a method of treating a patient having osteoporosis, the method comprising administering to the patient a therapeutically effective amount of a compound of Formula I or II.
  • the present invention provides a method of treating a patient having inflammation, the method comprising administering to the patient a therapeutically effective amount of a compound of Formula I or II.
  • the present invention provides a method of treating a patient having allergies, the method comprising administering to the patient a therapeutically effective amount of a compound of Formula I or II.
  • the present invention provides a method of treating a patient having cardiovascular disease, the method comprising administering to the patient a therapeutically effective amount of a compound of Formula I or II.
  • This invention provides compounds that inhibit the binding of proteins containing an SH2 domain with cognate phosphorylated proteins.
  • One group of compounds of the present invention have the Formula I
  • U is -CO- , -CS - , -SO- , or - S0 2 -
  • X is , or a bond
  • R 1 is hydrogen, RCONR'-, RR'NCONR -, RS0 2 NR'-, RCSNR'-, RR'NCSNR''-, RR'NS0 2 NR' ' - , ROCONR'-, or
  • R ⁇ is hydrogen, alkyl, cycloalkyl-(CH 2 ) n -, substituted alkyl, aryl-(CH 2 ) n _ , heteroaryl-(CH 2 ) n - -(CH 2 ) n -C0 2 H, substituted cycloalkyl-(CH ) n -/ substituted aryl-(CH 2 ) n -, or substituted heteroaryl-(CH 2 ) n -;
  • R 3 is hydrogen, alkyl, cycloalkyl- (CH 2 ) n ⁇ , substituted alkyl, aryl-(CH 2 ) n -, heteroaryl- (CH 2 ) n v - (CH 2 ) n -C0 2 H, substituted cycloalkyl-(CH 2 ) n ⁇ , substituted aryl-(CH 2 ) n _ , or substituted heteroaryl-(CH 2 ) n
  • R 5 is -NRR', — N ⁇ ' or -NCH(CH 3 )R;
  • R R and R 9 are independently -OP0 3 R c R d ,
  • R 7 and R 8 are independently hydrogen, alkyl, substituted alkyl, halogen, -OR, -NRR', -COCF 3 , -(CH 2 ) n CH 2 OH, -(CH 2 ) n C0 2 H, -(CH 2 ) n CHO, -(CH 2 ) n NRR', or -Q-CH 2 -(CH 2 ) n -NRR' ;
  • R 10 is -(CH 2 ) n C0 2 H, hydrogen, alkyl, aryl, substituted alkyl, or - (CH 2 ) n -substituted aryl;
  • R c and R d are independently -R, -CH 2 CH 2 Z, _ 0 0 R ,
  • Q is -0-, -NH-, -S-, -CH 2 0-, -CH 2 NH-, or -CH 2 S-;
  • Z is -Cl, -Br, or -F;
  • R, R' , and R' ' are independently hydrogen, alkyl, cycloalkyl- (CH 2 ) n - , aryl- (CH 2 ) n - , heteroaryl- (CH 2 ) n -/ substituted alkyl, substituted cycloalkyl- (CH 2 ) n _ substituted aryl- (CH 2 ) n _ , -(CH 2 ) n C0 H, or substituted heteroaryl- (CH 2 ) n - ; and each n is independently 0 to 5, or the pharmaceutically acceptable salts, amides, esters, or prodrugs thereof.
  • X is R 19 R 20 C > ⁇ a ⁇ or a bond
  • R 11 is hydrogen, alkyl, -OH, substituted alkyl, or
  • R 12 is hydrogen or alkyl
  • R 13 is -(CH 2 ) n C0 2 H, alkyl, -(CH 2 ) n -aryl, -(CH 2 ) n ⁇ heteroaryl, -(CH 2 ) n -cycloalkyl, hydrogen, substituted cycloalkyl-(CH 2 ) n - , substituted aryl-(CH 2 ) n -, substituted heteroaryl- (CH 2 ) n -, or substituted alkyl;
  • R 14 and R 17 are independently -OP0 3 R c R d , -CF 2 P0 3 R c R d , -CH 2 P0 3 R c R d , -P0 3 R c R d , -S0 3 R c , -OS0 3 R c , -CH 2 S0 3 R c , -S0 2 NH 2 , -OS0 2 NH 2 , or -CH 2 S0 2 NH 2 ;
  • R 15 and R 16 are independently hydrogen, alkyl, halogen, -OR, -NRR', -COCF 3 , -(CH 2 ) n CH 2 OH, -(CH 2 ) n C0 2 H, -(CH 2 ) n NRR', -(CH 2 ) n CHO, or -Q-CH 2 ⁇ (CH 2 ) n "NRR' ;
  • R 18 is -(CH 2 ) n C0 2 R, hydrogen, alkyl, -(CH 2 ) n CONRR' , substituted alkyl, or - (CH ) n -substituted aryl;
  • R 19 is hydrogen, RCONR'-, RR'NCONR''-, RS0 2 NR'-,
  • R 20 is hydrogen, alkyl, cycloalkyl-(CH ) n -, substituted alkyl, aryl-(CH 2 ) n -, heteroaryl-(CH 2 ) n -/ -(CH 2 ) n -C0 2 H, substituted cycloalkyl-(CH 2 ) n -, substituted aryl- (CH 2 ) n -, or substituted heteroaryl-(CH 2 ) n -;
  • R a is hydrogen, halogen, or alkyl;
  • R b is hydrogen, alkyl, -OR, -0(CH 2 ) n -aryl, -NRR', -0(CH 2 ) n -substituted alkyl-, -SR, -0(CH 2 ) n -substituted aryl, or -0(CH 2 ) n -cycloalkyl;
  • R c and R d are independently -R, -CH 2 CH Z, —o 0 R ,
  • Q is -0-, -NH-, -S-, -CH 2 0-, -CH 2 NH- , or -CH 2 S-;
  • Z is -Cl, -Br, or -F;
  • R, R', and R' ' are independently hydrogen, alkyl, cycloalkyl-(CH 2 ) n -, aryl-(CH ) n -, heteroaryl-(CH ) n -, -CH 2 -(CH 2 ) n -C0 2 H, substituted cycloalkyl-(CH 2 ) n -, substituted alkyl, substituted aryl-(CH 2 ) n -, or substituted heteroaryl-(CH 2 ) n -; and each n is independently 0 to 5, or the pharmaceutically acceptable salts, amides, esters, or prodrugs thereof.
  • alkyl means a straight or branched chain hydrocarbon.
  • the alkyl group has from 1 to 10 carbon atoms. More preferably, the alkyl group has from 1 to 6 carbon atoms. Examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, and tert-butyl.
  • cycloalkyl means a cyclic hydrocarbon, which can be saturated or unsaturated.
  • the cycloalkyl group has from 3 to 10 carbon atoms. More preferably, the cycloalkyl group has from 3 to 6 carbon atoms.
  • cycloalkyl groups include, but are not limited to, cyclopropane, cyclobutane, cyclopentane, cyclopentene, cyclohexane, cyclohexene, cycloheptane, cyclooctane, and adamantane.
  • aryl means a cyclic aromatic hydrocarbon.
  • aryl groups include, but are not limited to, phenyl and naphthyl.
  • heteroaryl means a cyclic aromatic hydrocarbon that contains one or more heteroatom. Examples of heteroatoms include, but are not limited to, oxygen, nitrogen, and sulfur. Examples of heteroaryl groups include, but are not limited to, pyridinyl, furanyl, thiophenyl, and pyrrolyl.
  • substituted aryl, substituted phenyl, substituted cycloalkyl, substituted heteroaryl, or substituted alkyl mean an aryl, phenyl, cycloalkyl, heteroaryl, or alkyl group that has one or more substituent.
  • substituents include alkyl, alkoxy, (such as methoxy, ethoxy, or tert-butoxy) , halogen, -N0 2 , -OCH 2 CONH 2 , -OCH 2 C0 2 H, -SO 3 H, -CONH 2 , -S0 2 NH 2 , or -CH 2 OH, and the like.
  • the symbol "-" means a bond.
  • proteins that contain an SH2 domain include, but are not limited to, Src, Fyn, Lck, Yes, Blk, Lyn, Fgr, Hck, Yrk, and Abl.
  • the protein is Src, and most preferably the protein is pp60c-src.
  • the term "cognate phosphorylated protein” means a protein to which the SH2 domain of a protein containing an SH2 domain binds or is associated.
  • cognate phosphorylated proteins include, but are not limited to, PDGF receptor protein, EGF receptor protein, HER2/Neu receptor protein (an oncogene form of EGF receptor protein), fibroblast growth factor receptor protein, focal adhesion kinase protein, pl30 protein, and p68 protein.
  • compositions suitable for parenteral injection may comprise physiologically acceptable sterile aqueous or nonaqueous solutions, dispersions, suspensions, or emulsions and sterile powders for reconstitution into sterile injectable solutions or dispersions.
  • aqueous and nonaqueous carriers, diluents, solvents or vehicles examples include water, ethanol, polyols (propyleneglycol, polyethyleneglycol, glycerol, and the like), suitable mixtures thereof, vegetable oils (such as olive oil), and injectable organic esters such as ethyl oleate.
  • a coating such as lecithin
  • surfactants for example
  • compositions may also contain adjuvants such as preserving, wetting, emulsifying, and dispensing agents.
  • adjuvants such as preserving, wetting, emulsifying, and dispensing agents.
  • Prevention of the action of microorganisms can be ensured by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, and the like.
  • isotonic agents for example, sugars, sodium chloride, and the like.
  • Prolonged absorption of the injectable pharmaceutical form can be brought about by the use of agents delaying absorption, for example, aluminum monostearate and gelatin.
  • Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules.
  • the active compound is admixed with at least one inert customary excipient (or carrier) such as sodium citrate or dicalcium phosphate or (a) fillers or extenders, as for example, starches, lactose, sucrose, glucose, mannitol, and silicic acid, (b) binders, as for example, carboxymethylcellulose, alignates, gelatin, polyvinylpyrrolidone, sucrose and acacia, (c) humectants, as for example, glycerol,
  • inert customary excipient such as sodium citrate or dicalcium phosphate
  • fillers or extenders as for example, starches, lactose, sucrose, glucose, mannitol, and silicic acid
  • binders as for example, carboxymethylcellulose, alignates, gelatin, polyvinylpyrrolidone, sucrose and acacia
  • humectants as for example, glycerol
  • disintegrating agents as for example, agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates and sodium carbonate,
  • absorption accelerators as for example, quaternary ammonium compounds
  • wetting agents as for example, cetyl alcohol and glycerol monostearate
  • adsorbents as for example, kaolin and bentonite
  • lubricants as for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof.
  • the dosage forms may also comprise buffering agents.
  • compositions of a similar type may also be employed as fillers in soft- and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethyleneglycols, and the like.
  • Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells, such as enteric coatings and others well known in the art. They may contain opacifying agents, and can also be of such composition that they release the active compound or compounds in a certain part of the intestinal tract in a delayed manner. Examples of embedding compositions which can be used are polymeric substances and waxes. The active compounds can also be in micro-encapsulated form, if appropriate, with one or more of the above-mentioned excipients.
  • Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs.
  • the liquid dosage forms may contain inert diluents commonly used in the art, such as water or other solvents, solubilizing agents and emulsifiers, as for example, ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propyleneglycol, 1,3-butyleneglycol, dimethylformamide, oils, in particular, cottonseed oil, groundnut oil, corn germ oil, olive oil, castor oil and sesame oil, glycerol, tetrahydrofurfuryl alcohol, polyethyleneglycols, and fatty acid esters of sorbitan or mixtures of these substances, and the like.
  • inert diluents commonly used in the art, such as water or other solvents, solubil
  • composition can also include adjuvants, such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.
  • adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.
  • Suspensions in addition to the active compounds, may contain suspending agents, as for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, or mixtures of these substances, and the like.
  • suspending agents as for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, or mixtures of these substances, and the like.
  • compositions for rectal administrations are preferably suppositories which can be prepared by mixing the compounds of the present invention with suitable non-irritating excipients or carriers such as cocoa butter, polyethyleneglycol or a suppository wax, which are solid at ordinary temperatures but liquid at body temperature and therefore, melt in the rectum or vaginal cavity and release the active component.
  • suitable non-irritating excipients or carriers such as cocoa butter, polyethyleneglycol or a suppository wax, which are solid at ordinary temperatures but liquid at body temperature and therefore, melt in the rectum or vaginal cavity and release the active component.
  • Dosage forms for topical administration of a compound of this invention include ointments, powders, sprays and inhalants.
  • the active component is admixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers or propellants as may be required.
  • Ophthalmic formulations, eye ointments, powders, and solutions are also contemplated as being within the scope of this invention.
  • pharmaceutically acceptable salts, esters, amides, and prodrugs refers to those carboxylate salts, amino acid addition salts, esters, amides and prodrugs of the compounds of the present invention which are, within the scope of sound medical judgement, suitable for use in contact with the tissues of patients without undue toxicity, irritation, allergic response, and the like, commensurate with a reasonable benefit/risk ratio, and effective for their intended use, as well as the zwitterionic forms, where possible, of the compounds of the invention.
  • salts refers to the relatively non-toxic, inorganic and organic acid addition salts of compounds of the present invention. These salts can be prepared in situ during the final isolation and purification of the compounds or by separately reacting the purified compound in its free base form with a suitable organic or inorganic acid and isolating the salt thus formed.
  • Representative salts include the hydrobro ide, hydrochloride, sulfate, bisulfate, nitrate, acetate, oxalate, valerate, oleate, palmitate, stearate, laurate, borate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, naphthylate mesylate, glucoheptonate, lactiobionate and laurylsulphonate salts, and the like.
  • alkali and alkaline earth metals such as sodium, lithium, potassium, calcium, magnesium, and the like
  • nontoxic ammonium, quaternary ammonium and amine cations including, but not limited to ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine, and the like.
  • ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine, and the like See, for example, Berge S.M., et al. , "Pharmaceutical Salts," J. Pharm. Sci. , 1977;66:1-19 which is incorporated herein by reference.
  • esters of the compounds of this invention examples include C- ⁇ -Cg alkyl esters wherein the alkyl group is a straight or branched chain. Acceptable esters also include C 5 -C 7 cycloalkyl esters as well as arylalkyl esters such as, but not limited to benzyl. C- ⁇ -C ⁇ j alkyl esters are preferred. Esters of the compounds of the present invention may be prepared according to conventional methods. Particularly preferred esters are phosphoesters. Examples of suitable phosphoesters include, but are not limited to -P0 3 R c R d ,
  • Examples of pharmaceutically acceptable, non-toxic amides of the compounds of this invention include amides derived from ammonia, primary C- ⁇ -Cg alkyl amines and secondary C j -C dialkyl amines wherein the alkyl groups are straight or branched chain. In the case of secondary amines, the amine may also be in the form of a 5- or 6-membered heterocycle containing one nitrogen atom. Amides derived from ammonia, C- ⁇ - ⁇ alkyl primary amides and C _ C 2 dialkyl secondary amides are preferred. Amides of the compounds of the invention may be prepared according to conventional methods.
  • the term "prodrug” refers to compounds that are rapidly transformed in vivo to yield the parent compound of the above formulas, for example, by hydrolysis in blood. A thorough discussion is provided in T. Higuchi and V. Stella, "Pro-drugs as Novel
  • the compounds of the present invention can be administered to a patient at dosage levels in the range of about 1 to about 7,000 mg/day. For a normal human adult having a body weight of about 70 kg, a dosage in the range of about 1 to about 100 mg/kg of body weight per day is sufficient.
  • the specific dosage used can vary. For example, the dosage can depend on a number of factors including the requirements of the patient, the severity of the condition being treated, and the pharmacological activity of the compound being used. The determination of optimum dosages for a particular patient is well known to those skilled in the art.
  • the compounds of the present invention can exist in different stereoisometric forms by virtue of the presence of asymmetric centers in the compounds. It is contemplated that all stereoisometric forms of the compounds as well as mixture thereof, including racemic mixtures, form part of this invention.
  • the compounds of the present invention can exist in unsolvated as well as solvated forms with pharmaceutically acceptable solvents such as water, ethanol, and the like. In general, the solvated forms are considered equivalent to the unsolvated forms for the purposes of the present invention.
  • the examples presented below are intended to illustrate particular embodiments of the invention and are not intended to limit the specification or the claims in any manner.
  • Step 1 Fmoc-L-Glu t 0tBu ⁇ -N tmethyl 3 -cyclohexyl - propyl )
  • Step 2 Ac -L-Tvr-L-Glu t OtBm -NfmethyH
  • Step 3 Ac-(O-phosphono ⁇ -L-Tyr-L-Glu-N tmethyl ⁇ (3-cyclohexylpropyl )
  • Boc- [4- (diethoxyphosphonyl) -difluoromethyl] - D/L-Phe Benzyl ester can be prepared in accordance with methods well known to those skilled in the art. (See, for example, Burke, et al . , J. Org. Chem. , 1993;58(6) :1336-1340. )
  • Step 2 Ac- f4- tdiethoxyphosphonyl ⁇ - difluoromethyll -D/L-Phe
  • Step 3 Ac- (4- (difluorophosphonomethyl ) ) -D/L-Phe- L-Glu-N(methyl (3-cvclohexvlpropvl>
  • Step 1 Ac- r 4 - ( diethoxyphosphonyl ⁇ - difluoromethy11- -Phe
  • Boc- [4-(diethoxyphosphonyl) -difluoromethyl] -L-Phe Benzyl ester can be prepared in accordance with methods well known to those skilled in the art. (See, for example, Smythe and Burke, Tett. Lett.. 1994;35(4) :551- 554. ) Boc- [4-(diethoxyphosphonyl)-difluoromethyl] -L- Phe Benzyl ester(1.75 mmol, 950 mg) was deprotected with 20 mL trifluoroacetic acid:dichloromethane (1:1) for 5 hours at 0°C.
  • the reaction was diluted with 200 mL of ethyl acetate and washed with saturated sodium bicarbonate then saturated sodium chloride, dried with sodium sulfate, filtered, and concentrated under reduced pressure to yield 795 mg of a colorless oil.
  • the crude oil was treated with acetic anhydride (10 mmol, 944 ⁇ L) and pyridine (15 mmol, 1.21 mL) in 20 mL dichloromethane for 4 hours at room temperature then 2 days at 4°C.
  • the title compound was synthesized in a manner similar to Example 4 only Ac- [4-(diethoxyphosphonyl) difluoromethyl] -L-Phe was coupled rather than the D/L mixture.
  • the purified peptide (30 mg) was deprotected with 1 M trimethysilyl triflate and 2 M dimethylsulfide in trifluoroacetic acid (3 mL) for 16 hours at room temperature. Water was added to quench excess trimethysilyl triflate, and the resulting solution was concentrated at reduced pressure to remove volatiles. The remaining solution was diluted with trifluoroacetic acid and water and purified by preparative HPLC, as previously described, to provide the product as a colorless solid after lyophilization (22 mg) .
  • Step 1 4 -Morpholinecarbon ⁇ l - L- Tyr t Bzl -OBzl
  • Step 4 4-Morpholinecarbonyl-(Q-phosphono -L-Tyr-
  • Step 2 N- r tMeth ⁇ lamino ⁇ carbonyl1 -L-Tyr-OH
  • N-methylcarbonyl-L-Tyr(Bzl)-OBzl 4.8 mmol, 2.0 g
  • methanol 50 mL
  • 10% palladium on carbon 0.50 g
  • the mixture was filtered over Celite, and the solvent was removed at the reduced pressure to give (1.27 g, 97%) of the product as an oil.
  • Step 3 N- r tMethvlamino ⁇ carbonyl 1 -L-T ⁇ r-L-
  • Step 1 Boc-L-Abu-N- (methyl 3-c ⁇ clohex ⁇ lprop ⁇ H Boc-L-Abu- (methyl) (3-cyclohexylpropyl) was synthesized in a manner similar to that described for Example 1 (Step 1) to yield a colorless oil.
  • the title compound was synthesized in a manner similar to that described for Example 1.
  • the product was obtained as a colorless solid (51 mg) .
  • HPLC 100%, rt 15.7 minutes, C18, eluting with a gradient of 10% to 76% acetonitrile containing 0.1% TFA, and water containing 0.1% TFA over 22 minutes.
  • Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 595.5 (M-H).
  • the title compound was synthesized in a manner similar to that described for Example 1.
  • the product was obtained as a colorless solid (51 mg) .
  • HPLC 100%, rt 13.3 minutes, C18 , eluting with a gradient of 10% to 76% acetonitrile containing 0.1% TFA, and water containing 0.1% TFA over 22 minutes.
  • Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 589.4 (M-H).
  • the title compound was synthesized in a manner similar to that described for Example 1.
  • the product was obtained as a colorless powder (52 mg) .
  • HPLC 100%, rt 14.0 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes.
  • Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 548.3 (M-H).
  • the title compound was synthesized in a manner similar to that described for Example 1.
  • the product was obtained as a colorless powder (35 mg) .
  • HPLC 100%, rt « 14.6 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes.
  • Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 571.5 (M-H).
  • the title compound was synthesized in a manner similar to that described for Example 1.
  • the product was obtained as a colorless powder (27 mg) .
  • HPLC 100%, rt 12.5 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes.
  • Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 534.5 (M-H).
  • Example i The title compound was synthesized in a manner similar to that described for Example i.
  • the product was obtained as a colorless powder (110 mg) .
  • HPLC 100%, rt 15.6 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes.
  • Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 584.3 (M-H).
  • the title compound was synthesized in a manner similar to that described for Example 1.
  • the product was obtained as a colorless powder (62 mg) .
  • HPLC 98%, rt 14.6 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes.
  • Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 579.4 (M-H).
  • the title compound was synthesized in a manner similar to that described for Example 1.
  • the product was obtained as a colorless powder (109 mg) .
  • HPLC 92%, rt 13.7 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes.
  • Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 548.4 (M-H).
  • Step 1 N-(t-ButylQxycarbpny )-2-f2- methoxyphenyl1-ethylamine
  • Step 3 Fmoc-L-Glu ( 0tBu 1 - N dneth ⁇ l H 2 - ( 2 - methoxyphenyl -ethyl) The compound was synthesized in a manner similar to that described for Example 1 (first step).
  • Step 4 Ac-L-Tyr-L-Glu(OtBu1-N(methyl 2-(2- methoxypheny11 -ethyl1
  • the compound was synthesized in a manner similar to that described for Example 1 (second step) .
  • Step 5 Ac- (0-phosphonol -L-T ⁇ r-L-Glu-N(meth ⁇ l1 ⁇ 2 -
  • Step 2 3- r3-(0-acetamido1 -phenyl) -propvl alcohol To 3- (3-hydroxyphenyl) -propyl alcohol (10.73 mmol,
  • Step 3 3- ⁇ 3 - (O-acetamido1 -phenvl) -propvl- (O- methanesulfonyl1
  • Step 4 3- 3-(0-acetamido> -phenvl) -propvl azide
  • Step 6 Fmoc-L-Glu(OtBu ) -NHf3- f3-(O-acetamido1 - pheny 1-propyl)
  • This compound was synthesized in a manner similar to that described for Example 1 (Step 1) .
  • Step 7 Ac-L-Tyr-L-Glu(OtBu1-NHf3- r3-(O- acetamido1-phenyl) ropyl)
  • Step 8 Ac- (O-phosphonol-L-Tyr-L-Glu-NHf3- ⁇ 3-(0- acetamido1-phenyl)propvl)
  • the title compound was synthesized in a manner similar to that described for Example 1. The product was obtained as a white solid (70 mg) .
  • HPLC 93%, rt 12.2 minutes, C18, eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes.
  • Electrospray Mass Spectrum 50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 622.4 (M-H).
  • the title compound was synthesized in a manner similar to that described for Example 1.
  • the product was obtained as a colorless powder (51 mg) .
  • Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 582.3 (M-H).
  • Aminobutanoate- gamma-tert-butyl ester can be prepared in accordance with methods well known to those skilled in the art. (See, for example, Sluka, et al. ,
  • Step 2 4-(3-Phenyl-propylaminol-butyric acid tert-butyl ester
  • the title compound was synthesized in a manner similar to that described for Example 1.
  • the product was obtained as a colorless powder (127 mg) .
  • HPLC 100%, rt 15.1 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes.
  • Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 576.7 (M-H).
  • the title compound was synthesized in a manner similar to that described for Example 1.
  • the product was obtained as a colorless powder (76 mg) .
  • HPLC 100%, rt 16.1 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes.
  • Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 590.3 (M-H).
  • the title compound was synthesized in a manner similar to that described for Example 1.
  • the product was obtained as a colorless powder (59 mg) .
  • HPLC 100%, rt 18.5 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes.
  • Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 547.4 (M-H).
  • the title compound was synthesized in a manner similar to that described for Example 1.
  • the product was obtained as a colorless solid (167 mg) .
  • HPLC 100%, rt 15.4 minutes, C18, eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes.
  • Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 575.5 (M-H).
  • EXAMPLE 32 4- r (RS1-2-(Acetyl-methvl-amino1-3- (4-phosphonooxy- phenyl)-(S)-propionylamino)-4 ⁇ (3 ⁇ phenyl- propylcarbamoyll-butyric acid or
  • the title compound was synthesized in a manner similar to that described for Example 1.
  • the product was obtained as a colorless powder (71 mg) .
  • HPLC 83%, rt 15.6 minutes, C18 column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes.
  • Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 584.3 (M-H).
  • EXAMPLE 34 rs-(R*,R*11-r4-(2-Acetylamino-2-fl-r(3-cyclohexyl' propyl1 -methyl-carbamoyl) -propylcarbamoy1) -ethyl1 ⁇ phenyl) -phosphonic acid or Ac-(4-phosphonvl1-L-Phe-L-Abu-N(methyl1 (3-cyclohexvlpropvl)
  • Step 1 Ac- f4-(diethoxyphosphonyl1) -L-Phe
  • Step 2 Ac- (4-phosphonyl1-L-Phe-L-Abu-N(methvl )
  • the title compound was synthesized in a manner similar to Example 10 except Ac- [4-(phosphonyl)- difluoromethyl]-L-Phe was coupled in rather than Ac- [4- (diethoxyphosphonyl)difluoromethyl] -L-Phe.
  • the purified peptide (30 mg) was deprotected with trimethysilyl bromide (1 mL) in dichloromethane (2 mL) for 4 hours at room temperature. Water (1 mL) and trifluoroacetic acid (1 mL) were added to quench excess trimethysilyl bromide, and the resulting solution was concentrated at reduced pressure to remove volatiles.
  • the isocyanate was treated in dichloromethane (20 mL) with 4-hydroxybenzyl glycine-t-butyl ester (250 mg, 1.0 mmol) and triethylamine (0.15 mL, 1.0 mmol) for 1 hour. Ethyl acetate was added, and the reaction mixture was washed with 10% sulfuric acid, then water, and then brine. After drying over magnesium sulfate, the solvent was removed under reduced pressure to give then urea as a colorless oil (500 mg, 83%).
  • Step 1 (S1-2-r3-(6-Hydroxy-naphthalen-l-yl1- ureido) -pentanedioic acid 5-tert-butvl ester 1-methyl ester
  • Step 3 (S )-4- f (3-Cyclohexyl-propyl1- ethyl- carbamoyl) -4- T3-(6-hydroxv-naphthalen-l-yll-ureido) - butyric acid tert-butvl ester
  • This step was synthesized 'in a manner similar to that described for Example 1 (Step 1) .
  • Step 4 (SI-4- (3-Cyclohexyl-propyl -methyl- carbamoyl) -4- ⁇ 3 -(6-phosphonox ⁇ -naphthalen-l-yl ureidol-butyric acid
  • Example 44 The title compound was synthesized in a manner similar to that described for Example 44.
  • the product was obtained as a colorless powder (21 mg) .
  • HPLC 100%, rt 14.8 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes.
  • Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 455.2 (M-H).
  • Example 44 The title compound was synthesized in a manner similar to that described for Example 44.
  • the product was obtained as a colorless powder (73 mg) .
  • HPLC 100%, rt 16.9 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes.
  • Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 469.3 (M-H).
  • Example 44 The title compound was synthesized in a manner similar to that described for Example 44.
  • the product was obtained as a colorless powder (85 mg) .
  • HPLC 100%, rt 17.2 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes.
  • Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 483.4 (M-H).
  • the title compound was synthesized in a manner similar to that described for Example 44.
  • the product was obtained as a colorless powder (8 mg) .
  • HPLC 100%, rt 18.7 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes.
  • Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 449.4 (M-H).
  • Example 44 The title compound was synthesized in a manner similar to that described for Example 44.
  • the product was obtained as a colorless powder (100 mg) .
  • HPLC 99%, rt 18.3 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes.
  • Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 463.4 (M-H).
  • Step 1; 5-Aminomethvlsalicvlic acid 5-Aminomethylsalicylic acid can prepared in accordance with methods well known to those skilled in the art. (See, for example, Sekiya, et al., Chem. Pharm. Bull. , 1963;11:551-553.)
  • the reaction was then concentrated under reduced pressure. The residue was diluted with ethyl acetate and 5% citric acid. The organic layer was washed with 5% citric acid (2X50 mL) , IN sodium hydroxide (2X50 mL) , 5% citric acid (2X50 mL) , and saturated sodium chloride (1X50 mL), dried over magnesium sulfate, filtered, and concentrated under reduced pressure to yield a pale yellow powder. The powder was collected and washed with ether, followed by drying under reduced pressure to yield (1.5 g, 57%).
  • Step 5 Benzamide, 2-(cyclohexylmethoxy )-5- T f f (4- hvdroxvphenvl1-carbonvl1 mino) -methyl)
  • a solution of 4N hydrochloric acid in dioxane was added to (3-Carbamoyl-4-cyclohexylmethoxy-benzyl) - carbamic acid tert-butyl ester (1.0 g, 2.7 mmol). The reaction is stirred for 1 hour and then concentrated under reduced pressure to yield a solid. This solid of the amine hydrochloride was then used for the coupling.
  • Step 6 Phosphoric acid mono- -(3-carbamoyl-4- cyclohexylmethoxy-benzvlcarbamovl1 -phenyl) ester
  • the title compound was synthesized in a manner similar to that described for Example 1.
  • the product was obtained as a colorless powder (50 mg) .
  • HPLC 92%, rt 17.2 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes.
  • Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 461.5 (M-H).
  • Step 1 fPifluoro-(4-iodo-phenyll-methyl) - phosphonic acid diethyl ester
  • [Difluoro-(4-iodo-phenyl)-methyl] -phosphonic acid diethyl ester can be prepared in accordance with methods well known to those skilled in the art. (See, for example, Burke, et al. , J. Org. Chem. , 1993;58: 1336-1340. )
  • Step 2 fDifluoro- (4-formvl-phenyl )-methyl) - phosphonic acid diethyl ester
  • n-butyl lithium 2.5 M in hexanes, 2.5 mL, 6.1 mmol
  • Ethyl formate (0.66 mL, 8.2 mmol) was then added, and the reaction was stirred for 10 minutes at -78°C.
  • the mixture was then quenched with saturated ammonium chloride (10 mL) and warmed to room temperature.
  • the reaction was then diluted with ether.
  • the organic layers were washed with saturated sodium chloride
  • Step 3 4- ⁇ (Diethoxy-phosphoryl)-difluoro ⁇ methyl) -benzoic acid
  • acetone 25 mL
  • Jones reagent 5 mL
  • ethanol 5 mL
  • the reaction was then concentrated under reduced pressure to 5 mL.
  • Step 4 r4- (3-Carbamoyl-4-cyclohexylmethoxy- benzylcarbamoyll -phenyl) -difluoro-methyl-phosphonic acid
  • Example 44 The title compound was synthesized in a manner similar to that described for Example 44.
  • the product was obtained as a colorless powder (39 mg) .
  • HPLC 100%, rt 17.4 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes.
  • Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 495.4 (M-H).
  • Example 44 The title compound was synthesized in a manner similar to that described for Example 44.
  • the product was obtained as a colorless powder (79 mg) .
  • HPLC 100%, rt 18.3 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes.
  • Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 489.4 (M-H).
  • Example 44 The title compound was synthesized in a manner similar to that described for Example 44.
  • the product was obtained as a colorless powder (119 mg) .
  • HPLC 100%, rt 17.7 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes.
  • Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 495.3 (M-H).
  • Example 44 The title compound was synthesized in a manner similar to that described for Example 44.
  • the product was obtained as a colorless powder (79 mg) .
  • HPLC 100%, rt 18.3 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes.
  • Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 475.5 (M-H).
  • the title compound was synthesized in a manner similar to that described for Example 44.
  • the product was obtained as a colorless powder (60 mg) .
  • HPLC 99%, rt 16.9 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes.
  • Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 546.4 (M-H).
  • Example 44 The title compound was synthesized in a manner similar to that described for Example 44.
  • the product was obtained as a colorless powder (59 mg) .
  • HPLC 98%, rt 17.0 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes.
  • Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 568.4 (M-H).
  • the title compound was synthesized in a manner similar to that described for Example 44.
  • the product was obtained as a colorless powder (61 mg) .
  • HPLC 99%, rt 19.7 and 20.10 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes.
  • Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 560.4 (M-H).
  • the title compound was synthesized in a manner similar to that described for Example 44.
  • the product was obtained as a colorless powder (61 mg) .
  • HPLC 99%, rt 19.7 and 20.1 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes.
  • Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 560.4 (M-H).
  • Step 1 (S1-3-(4-Benzyloxy-phenyl1-2-(3-methyl- ureido)-propionic acid benzyl ester This compound was synthesized in a manner similar to that described in Example 7 (Step 1).
  • This compound was synthesized in a manner similar to that described in Example 7 (Step 2) .
  • Step 3 (S )-2-Cyclohexylmethoxy-5-f ⁇ 3-(4-hydroxy ⁇ phenyl1-2-(3-methvl-ureidol-propionylamino) -methyl)- benaamide
  • Step 4 (SI-Phosphoric acid mono-f4-r2- acetylureido-2-(3-carbamovl-4- cyclohexylmethoxvbenzyIcarbamovl1-ethvl) -phenvl)ester
  • Product was obtained as a white solid (100 mg) .
  • HPLC 98%, rt 17.4 minutes, C18, eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes.
  • Electrospray Mass Spectrum 50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 561.4 (M-H).
  • the title compound was synthesized in a manner similar to that described for Example 34.
  • the product was obtained as a colorless powder (125 mg) .
  • HPLC 100%, rt 12.4 minutes, C18, column eluting with a gradient of 0% to 100% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 20 minutes.
  • Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 552 (M-H).
  • N(methyl1 (3-cyclohexylpropyl ) (from Example 10) (0.15 mmol, 84 mg) was dissolved in 2 mL dimethylformamide and treated with diisopropylethylamine (0.5 mmol, 81 ⁇ L) and chloromethyl pivaloate (1.5 mmol, 214 ⁇ L) .
  • the reaction mixture was stirred at 70 °C for 24 hours.
  • the reaction mixture was diluted with 3 mL of dimethylformamide and purified by preparative HPLC, as previously described, to provide the product as a colorless solid after lyophilization (58 mg) .
  • the title compound was synthesized in a manner similar to that described for Example 1.
  • the product was obtained as a white solid (50 mg) .
  • HPLC 100%, rt 18.0 minutes, C18, eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes.
  • Electrospray Mass Spectrum (50/50 acetonitrile/water +0.1% ammonium hydroxide) m/z 593 (M-H) .
  • Example 51 The title compound is the second isomer separated in Example 51 and 53.
  • the product was obtained as a colorless powder (34 mg) .
  • HPLC >95%, rt 12.8 minutes, C18, column eluting with a gradient of 0% to 100% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 20 minutes.
  • Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 594 (M-H).
  • EXAMPLE 64b Phosphoric acid mono- T4-(2-acetylamino-2-f1- f3- carbamoy1-4-(2-cyclohexyl-1-methvl-ethoxy )- henyl1 - ethvlcarbamoyl)-ethyl )-phenvl) ester
  • Step 2 (2-methoxyphenyl -acetic acid methvl ester
  • Step 4 (5-acetyl-2-cyclohexylmethoxyphenyl)- acetic acid methvl ester This compound was synthesized in a manner similar to that described for Example 44, Step 4. The product was obtained as an oil (7.9 g) .
  • Mass Spectrum (Chemical Ionization, 1% NH 3 in CH 4 ) m/z 305 (M+H).
  • Step 5 ( 5-Acetvl - 2-cvclohexylmethoxy phenyl ) - acetic acid
  • a solution of (5-Acetyl-2-cyclohexylmethoxy phenyl)-acetic acid methyl (7.9 g, 25.9 mmol) in tetrahydrofuran (150 mL) and methanol (150 mL) was added a IN solution of sodium hydroxide (52.0 mL, 52 mmol). The reaction was stirred at room temperature overnight. The reaction was then concentrated under reduced pressure, diluted with ethyl acetate and acidified to pH 2 with IN hydrochloric acid.
  • step 6 2-( -Acetyl- -cyclohexyl ethoxy- henyl)- acetamide
  • Example 44 The title compound was synthesized in a manner similar to that described for Example 44.
  • the product was obtained as a colorless powder (93 mg) .
  • HPLC 100%, rt 17.8 and 18.0 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes.
  • Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 574 (M-H).
  • Example 44 The title compound was synthesized in a manner similar to that described in Example 44.
  • the product was obtained as a colorless solid (101 mg) .
  • HPLC 98%, rt 18.9 minutes, C18, eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA, and water containing 0.1% TFA over 22 minutes.
  • Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 574 (M-H) .
  • Example 65 The title compound was synthesized in a manner similar to that described for Example 65 starting at step 3. The product was obtained as a colorless powder (26 mg) .
  • HPLC 100%, rt 20.2 and 20.5 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes.
  • Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 636 (M-H) .
  • Example 51 The title compound was synthesized in a manner similar to that described for Example 51.
  • the product was obtained as a colorless powder (16 mg) .
  • HPLC 90%, rt 16.9 and 17.3 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes.
  • Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 582 (M-H).
  • Example 65 The title compound was synthesized in a manner similar to that described for Example 65 starting at step 3. The product was obtained as a colorless powder (32 mg) .
  • Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 588 (M-H) .
  • Step 1 l-isocyanato-4-(phenvlmethoxy)benzene To amine (21.20 mmol, 5.0 g) in toluene (10 ml) at room temperature was added phosgene in toluene (1.93 M, 106 mmol, 54 mL) . After refluxing 12 hours, the mixture was evaporated to dryness under reduced pressure. The crude product was used directly in the next step without further purification or characterization.
  • This compound was synthesized in a manner similar to that described in Example 7 (step 1) .
  • Step 3 2-Cvclohexylmethoxy5- T3- (4-hydroxy ⁇ phenyl)-ureidomethvl) -benzamide This compound was synthesized in a manner similar to that described in Example 6 (step 2) .
  • Step 4 Phosphoric acid mono-f4- ⁇ 3-(3-carbamoyl-4- cyclohexylmethoxybenzyl)-ureido) -phenyl) ester
  • Example 44 step 5 The title compound was synthesized in a manner similar to that described in Example 35 only the amine hydrochloride produced in Example 44 step 5 was employed as a partner in the urea forming reaction. The product was obtained as a colorless solid (78 mg) .
  • HPLC 98%, rt 17.5 minutes, C18, eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA, and water containing 0.1% TFA over 22 minutes.
  • Electrospray Mass Spectrum 50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 490 (M-H).
  • the binding affinities of compounds of the present invention to Src SH2 was determined using a competitive radiolabeled phosphopeptide displacement assay. Specifically, binding of 125 I-labeled Glu-Pro-Gln-pTyr- Glu-Glu-Ile-Pro-Ile-Tyr-Leu or 125 I-labeled Glu-Pro- Gln- (4-(difluorophosphonomethyl) )-Phe-Glu-Glu-Ile-Pro- Ile-Tyr-Leu to a glutathione-S-transferase(GST) -Src SH2 fusion protein was performed in 20 mM Tris (pH 7.5), 150 mM NaCl, 5 mM EDTA, and 0.1% NP-40.
  • Immobili ed Abl SH2 The binding affinities of compounds of the present invention to Abl SH2 was determined using a competitive radiolabeled phosphopeptide displacement assay. Specifically, binding of 125 I-labeled Glu-Pro-Gln- (4- (difluorophosphonomethyl) )-Phe-Glu-Glu-Ile-Pro-Ile-Tyr- Leu to a glutathione-S-transferase(GST)-Abl SH2 fusion protein was performed in 20 mM Tris (pH 7.5), 150 mM NaCl, 5 mM EDTA, and 0.1% NP-40.
  • DNA synthesis occurs when a cell is exposed to Platelet Derived Growth Factor (PDGF) .
  • PDGF Platelet Derived Growth Factor
  • c-SRC c-SRC or another member of the SRC gene family is required.
  • PDGF-stimulated DNA synthesis can be assessed using the protocol set forth below.
  • Swiss 3T3 cells were grown in 12-well plates for 3 days to approximately 50% confluency. The growth media was removed and replaced with 1.0 mL/well of assay buffer (Dulbecco's Modified Eagle Medium containing 0.2% Bovine Serum Albumin), and incubated for 24 hours to arrest growth.
  • assay buffer Dulbecco's Modified Eagle Medium containing 0.2% Bovine Serum Albumin
  • Test compounds (5 ⁇ L) were add at 100 times their final concentration. Methyl sulfoxide was used as a control and levels in all wells are kept under 1.0% of DMSO. Cells were then stimulated with growth factor (PDGFbb or Fetal Bovine Serum), and allowed to incubate another 24 hours.
  • PDGFbb or Fetal Bovine Serum growth factor

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Abstract

The present invention provides compounds that inhibit the binding of proteins containing an SH2 domain to cognate phosphorylated proteins. The invention also provides pharmaceutical compositions containing the compounds and methods of inhibiting the binding of proteins containing an SH2 domain to cognate phosphorylated proteins.

Description

COMPOUNDS, COMPOSITIONS AND METHODS FOR INHIBITING THE
BINDING OF PROTEINS CONTAINING AN SH2 DOMAIN
TO COGNATE PHOSPHORYLATED PROTEINS
FIELD OF THE INVENTION
This invention relates to compounds that inhibit the binding of proteins containing an SH2 domain to cognate phosphorylated proteins. This invention also relates to pharmaceutical compositions containing the compounds and to therapeutic methods that use the compounds.
BACKGROUND OF THE INVENTION
Many of the signal transduction pathways that regulate a variety of cellular processes, including the differentiation and proliferation of normal and malignant cells, operate through phosphorylated proteins called tyrosine kinases. The two major types of tyrosine kinases are receptor and nonreceptor tyrosine kinases. Receptor tyrosine kinases contain binding sites or receptors for growth factors such as epidermal growth factor (EGF) or platelet-derived growth factor (PDGF) . When a growth factor such as EGF or PDGF binds to the receptor, the tyrosine kinase receptor protein is activated; the tyrosine kinase receptor protein is autophosphorylated; and endogenous proteins that participate in the signal transduction pathway are phosphorlyated.
Some endogenous proteins that are involved in the cellular signal transduction pathways contain a specific domain called the SH2 domain, which provides for interaction with an activated tyrosine kinase receptor protein. This interaction is a protein- protein interaction. One protein that contains an SH2 domain is pp60c-src kinase, which is also a tyrosine kinase. The pp60c-src kinase is a nonreceptor kinase and is related to a number of nonreceptor tyrosine kinases, which include, but are not limited to, Fyn, Lck, Yes, Blk, Lyn, Fgr, Hck, and Yrk. Another important protein that contains an SH2 domain is Abl. The pp60c-src protein is exemplary of the Src family of tyrosine kinases. The pp60c-src has three major domains: SHI, SH2, and SH3. The SHI domain is most commonly called the catalytic domain or tyrosine kinase domain. The SH3 domain is a binding region for proteins having proline-rich sequences. Both the SH2 and SH3 domains are noncatalytic, but are important in protein-protein recognition.
The Src family of protein kinases, which all contain a SH2 domain, are involved in a number of cellular signalling pathways. For example, Src is involved in growth factor receptor signalling; integrin-mediated signaling; T- and B-cell activation and osteoclast activation. It is known that the Src SH2 domain binds to several key receptor and nonreceptor tyrosine kinases such as tyrosine kinases containing receptors for PDGF, EGF, HER2/Neu (an oncogene form of EGF), Fibroblast growth factor, focal adhesion kinase, pl30 protein, and p68 protein. In addition, pp60c-src has been shown to be involved in the regulation of DNA synthesis, mitosis, and other cellular activities.
Thus, it would be useful to have compounds that inhibit the binding of proteins containing an SH2 domain to cognate phosphorylated proteins, as the inhibition of binding of proteins containing an SH2 domain to cognate phosphorylated proteins can be used to treat proliferative diseases such as cancer, osteoporosis, inflammation, allergy, restenosis, and cardiovascular disease, which all rely on signal transduction involving proteins that contain an SH2 domain that binds to phosphorylated proteins during the cellular signalling process.
SUMMARY OF THE INVENTION The present invention provides compounds having the Formula I
Figure imgf000005_0001
wherein
U is -CO-, -CS-, -SO-, or -S02' Y is
Figure imgf000005_0002
Figure imgf000005_0003
X is or a bond;
Figure imgf000006_0001
R1 is hydrogen, RCONR'-, RR'NCONR''-, RS02NR'-, RCSNR'-, RR'NCSNR''-, RR'NS02NR' ' - , ROCONR'-, or . .
0 NCONR' - ; \ /
R2 is hydrogen, alkyl, cycloalkyl- (CH2)n- , substituted alkyl, aryl- (CH2)n- , heteroaryl- (CH2)n- , - (CH2)n-C02H, substituted cycloalkyl- (CH2)n- , substituted aryl- (CH2)n- or substituted heteroaryl- (CH2)n _ ;
R3 is hydrogen, alkyl, cycloalkyl- (CH2)n- , substituted alkyl, aryl- (CH2)n- , heteroaryl- (CH2)n- , -(CH2)n-C02H, substituted cycloalkyl- (CH2)n-, substituted aryl- (CH2)n- , or substituted heteroaryl- (CH2)n- ;
R4 is hydrogen or alkyl;
R'
>5 i.js_ --NXTROR-D'',r ——NN O^ t or -NCH(CH3)R;
R6 and R9 are independently -OP03RcRd, -CF2P03RcRd, -CH2P03RcRd, -P03RcRd, -S03Rc, -OS03Rc, -CH2S03Rc, -S02NH2, -OS02NH2, or -CH2S02NH2;
R 7' and Rft° are independently hydrogen, alkyl, substituted alkyl, halogen, -OR, -NRRX -COCF3, -(CH2)nCH2OH, -(CH2)nC02H, -(CH2)nCHO, -(CH2)nNRRX or -Q-CH2-(CH2)n-NRR'; R10 is -(CH2)nC02H, hydrogen, alkyl, aryl, substituted alkyl, or - (CH2)n-substituted aryl;
Rc and Rd are independently -R, -CH2CH2Z,
-CH2CHZ2, -CH2CZ3, or -°vI °vH R' ; R O
Q is -0-, -NH-, -S-, -CH20-, -CH2NH-, or -CH2S-;
Z is -Cl, -Br, or -F; R, R', and R' ' are independently hydrogen, alkyl, cycloalkyl-(CH2)n-, aryl-(CH2)n-, heteroaryl-(CH2)n", substituted alkyl, substituted cycloalkyl-(CH2)n _, substituted aryl- (CH2)n _, -(CH2)nC0 H, or substituted heteroaryl-(CH2)n-; and each n is independently 0 to 5, or the pharmaceutically acceptable salts, amides, esters, or prodrugs thereof.
In a preferred embodiment of the compounds of Formula I,
U is -CO-
Figure imgf000007_0001
In a preferred embodiment of the compounds of Formula I,
U is -CO-
,lτ,2 /
X is R-^c and
\
Figure imgf000007_0002
R1 is RCONR'-, -NRCONR'R'X -NRS02RX or
NCONR' -; R2, R4, R7, and R8 are hydrogen;
R3 is -(CH2)nC02H, alkyl, or - (CH2)n-substituted aryl;
R' is -NRR', -NCH(CH )R, or
~N\ ; and
R
R6 is -0P03RcRd, -CF2P03RcRd, or -P03RcRd. In another preferred embodiment of the compounds of Formula I
U is -CO-;
/
X is R1R2C and \
Figure imgf000008_0001
Figure imgf000008_0002
R8 are hydrogen; R3 is -CH2CH2C02H;
R5 is -NRR', - ; and
Figure imgf000008_0003
R R6 is -OP03RcRd, -CF2P03RcRd, or -P03RcRd. In another preferred embodiment of the compounds of Formula I
Figure imgf000008_0004
Figure imgf000009_0001
In another preferred embodiment of the compounds of Formula I
U is -C0-
Figure imgf000009_0002
R3 and R10 are -(CH2)nC02H; R4 is hydrogen;
Figure imgf000009_0003
R7 and R8 are hydrogen; and R6 is -OP03RcRd, -CF2P03RcRd, or -P03RcRd. In another aspect, the present invention provides the compounds of Formula II below:
Figure imgf000009_0004
wherein U and W are independently -CO-, -CS-, -SO-, or -S02-;
Figure imgf000010_0001
Figure imgf000010_0002
Figure imgf000010_0003
/ is R19R20C >1-L -N or a bond;
\
\
R 11 is hydrogen, alkyl, -OH, substituted alkyl, or
■ NH-
R- 1*- is hydrogen or alkyl;
R13 is -(CH2)nC02H, alkyl, - (CH2)n-aryl, - (CH2)n-heteroaryl, - (CH2)n-cycloalkyl, hydrogen, substituted cycloalkyl- (CH2)n- , substituted aryl- (CH2)n- , substituted heteroaryl- (CH2)n- , or substituted alkyl;
R14 and R17 are independently -OP03RcRd, -CF2P03RcRd, -CH2P03RcRd, -P03RcRd, -S03Rc, -OS03Rc, -CH2S03Rc, -S02NH2, -OS02NH2, or -CH2S02NH2;
R15 and R16 are independently hydrogen, alkyl, halogen, -OR, -NRRX -COCF3, - (CH2)nCH2OH, -(CH2)nC02H, -(CH2)nNRRX -(CH2)nCHO, or -Q-CH2" (CH2)n-NRR' ;
R18 is -(CH2)nC02R, hydrogen, alkyl, - (CH2)nCONRR' , substituted alkyl, or - (CH2)n-substituted aryl; R19 is hydrogen, RCONR'-, RR'NCONR''-, RS02NR'
RR'NS02NR' '-, ROCONR'-, or o NCONR'- ;
R20 is hydrogen, alkyl, cycloalkyl- (CH2)n- / substituted alkyl, aryl- (CH2)n" , heteroaryl- (CH2)n-, - (CH2)n-C02H, substituted cycloalkyl- (CH2)n- , substituted aryl- (CH2)n~ , or substituted heteroaryl- (CH2)n~; Ra is hydrogen, halogen, or alkyl;
Rb is hydrogen, alkyl, -OR, -0(CH2)n-aryl, -NRRX -0(CH2)n-substituted alkyl, -SR, -0(CH2)n-substituted aryl, or -0(CH2)n-cycloalkyl;
Rc and Rd are independently -R, -CH2CH2Z, —o 0 R,
-CH2CHZ2, -CH2CZ3, or j fl
R 0
Q is -0-, -NH-, -S-, -CH20-, -CH2NH-, or -CH2S-;
Z is -Cl, -Br, or -F; R, RX and R' ' are independently hydrogen, alkyl, cycloalkyl- (CH2)n- , aryl- (CH2)n- , heteroaryl- (CH2)n- , -CH2-(CH2)n-C02H, substituted cycloalkyl- (CH2)n- , substituted alkyl, substituted aryl- (CH )n- , or substituted heteroaryl- (CH2)n- ; and each n is independently 0 to 5, or the pharmaceutically acceptable salts, amides, esters, or prodrugs thereof.
In a preferred embodiment of the compounds of Formu
Figure imgf000011_0001
U and W are -CO- ; and X is a bond. In another preferred embodiment of the compounds of Fo
Figure imgf000012_0001
U and W are -CO-; X is a bond;
R ,1±3J, Rα,
Figure imgf000012_0002
, and R 1±6Ό are hydrogen;
R and R4--1- are hydrogen or alkyl;
Rb is -OR, -0(CH2)n-aryl, -0(CH2)n substituted aryl, or -0(CH2)ncycloalkyl; and
,14 -OP02R >Cv-τR_du or -CF2P03Rc"-DRd In another preferred embodiment of the compounds of Formula II,
/
X is R19R 0c \
Figure imgf000012_0003
U and W are -CO- .
In another preferred embodiment of the compounds of Formula II,
/
X is R19R20C \
Figure imgf000012_0004
U and W are -CO- ;
R19 is RCONR ' - or RR ' NCONR ' ' - ;
R20, R15, R13, R11, Ra, and R16 are hydrogen;
R12 is alkyl or hydrogen; Rb is -OR, -0(CH2)n-aryl, -0(CH2)n substituted aryl, or -0(CH2)ncycloalkyl; and
R14 is -OP03RcRd or -CF2P03RcRd.
Also provided by the present invention is method of inhibiting the binding of a protein containing an SH2 domain to a cognate phosphorylated protein, the method comprising administering to a patient in need of SH2 inhibition an SH2 inhibiting amount of a compound of Formula I or II. In another aspect, the present invention provides a pharmaceutical composition that comprises a compound of Formula I or II and a pharmaceutically acceptable carrier.
In another aspect, the present invention provides a method of treating a patient having a proliferative disease, the method comprising administering to the patient a therapeutically effective amount of a compound of Formula I or II.
In another aspect, the present invention provides a method of treating a patient having cancer, the method comprising administering to the patient a therapeutically effective amourit of a compound of Formula I or II.
In another aspect, the present invention provides a method of treating a patient having restenosis, the method comprising administering to the patient a therapeutically effective amount of a compound of Formula I or II.
In another aspect, the present invention provides a method of treating a patient having osteoporosis, the method comprising administering to the patient a therapeutically effective amount of a compound of Formula I or II.
In another aspect, the present invention provides a method of treating a patient having inflammation, the method comprising administering to the patient a therapeutically effective amount of a compound of Formula I or II.
In another aspect, the present invention provides a method of treating a patient having allergies, the method comprising administering to the patient a therapeutically effective amount of a compound of Formula I or II.
In another aspect, the present invention provides a method of treating a patient having cardiovascular disease, the method comprising administering to the patient a therapeutically effective amount of a compound of Formula I or II.
DETAILED DESCRIPTION OF THE INVENTION
This invention provides compounds that inhibit the binding of proteins containing an SH2 domain with cognate phosphorylated proteins. One group of compounds of the present invention have the Formula I
Figure imgf000014_0001
wherein
U is -CO- , -CS - , -SO- , or - S02 -
Y is
Figure imgf000015_0001
X is , or a bond;
Figure imgf000015_0002
R1 is hydrogen, RCONR'-, RR'NCONR -, RS02NR'-, RCSNR'-, RR'NCSNR''-, RR'NS02NR' ' - , ROCONR'-, or
NCON '
R^ is hydrogen, alkyl, cycloalkyl-(CH2)n-, substituted alkyl, aryl-(CH2)n _, heteroaryl-(CH2)n- -(CH2)n-C02H, substituted cycloalkyl-(CH )n-/ substituted aryl-(CH2)n-, or substituted heteroaryl-(CH2)n-; R3 is hydrogen, alkyl, cycloalkyl- (CH2)n~, substituted alkyl, aryl-(CH2)n-, heteroaryl- (CH2)nv - (CH2)n-C02H, substituted cycloalkyl-(CH2)n~, substituted aryl-(CH2)n _, or substituted heteroaryl-(CH2)n-; R4 is hydrogen or alkyl; R'
R5 is -NRR', —N\ ' or -NCH(CH3)R;
R R and R9 are independently -OP03RcRd,
-CF2P03RcRd, -CH2P03RcRd, -P03RcRd, -S03Rc, -OS03Rc, -CH2S03Rc, -S02NH2, -OS02NH2, or -CH2S02NH2;
R7 and R8 are independently hydrogen, alkyl, substituted alkyl, halogen, -OR, -NRR', -COCF3, -(CH2)nCH2OH, -(CH2)nC02H, -(CH2)nCHO, -(CH2)nNRR', or -Q-CH2-(CH2)n-NRR' ;
R10 is -(CH2)nC02H, hydrogen, alkyl, aryl, substituted alkyl, or - (CH2)n-substituted aryl; Rc and Rd are independently -R, -CH2CH2Z, _0 0 R,
-CH2CHZ2, -CH2CZ3, or | H ;
R O
Q is -0-, -NH-, -S-, -CH20-, -CH2NH-, or -CH2S-;
Z is -Cl, -Br, or -F; R, R' , and R' ' are independently hydrogen, alkyl, cycloalkyl- (CH2)n- , aryl- (CH2)n- , heteroaryl- (CH2)n-/ substituted alkyl, substituted cycloalkyl- (CH2)n _ substituted aryl- (CH2)n _ , -(CH2)nC0 H, or substituted heteroaryl- (CH2)n- ; and each n is independently 0 to 5, or the pharmaceutically acceptable salts, amides, esters, or prodrugs thereof.
Another group has the Formula II below:
Figure imgf000016_0001
wherein U and W are independently -CO-, -CS-, -SO-, or -S02-; Y is
Figure imgf000017_0001
Figure imgf000017_0002
/ /
X is R19R20C >ιa ■ or a bond; \
\
R 11 is hydrogen, alkyl, -OH, substituted alkyl, or
• NH-
R12 is hydrogen or alkyl;
R13 is -(CH2)nC02H, alkyl, -(CH2)n-aryl, -(CH2)n~heteroaryl, -(CH2)n-cycloalkyl, hydrogen, substituted cycloalkyl-(CH2)n- , substituted aryl-(CH2)n-, substituted heteroaryl- (CH2)n-, or substituted alkyl;
R14 and R17 are independently -OP03RcRd, -CF2P03RcRd, -CH2P03RcRd, -P03RcRd, -S03Rc, -OS03Rc, -CH2S03Rc, -S02NH2, -OS02NH2, or -CH2S02NH2;
R15 and R16 are independently hydrogen, alkyl, halogen, -OR, -NRR', -COCF3, -(CH2)nCH2OH, -(CH2)nC02H, -(CH2)nNRR', -(CH2)nCHO, or -Q-CH2~(CH2)n"NRR' ;
R18 is -(CH2)nC02R, hydrogen, alkyl, -(CH2)nCONRR' , substituted alkyl, or - (CH )n-substituted aryl; R19 is hydrogen, RCONR'-, RR'NCONR''-, RS02NR'-,
RR'NS02NR' '-, ROCONR'-, or 0 NCONR'- !
R20 is hydrogen, alkyl, cycloalkyl-(CH )n-, substituted alkyl, aryl-(CH2)n-, heteroaryl-(CH2)n-/ -(CH2)n-C02H, substituted cycloalkyl-(CH2)n-, substituted aryl- (CH2)n-, or substituted heteroaryl-(CH2)n-; Ra is hydrogen, halogen, or alkyl;
Rb is hydrogen, alkyl, -OR, -0(CH2)n-aryl, -NRR', -0(CH2)n-substituted alkyl-, -SR, -0(CH2)n-substituted aryl, or -0(CH2)n-cycloalkyl;
Rc and Rd are independently -R, -CH2CH Z, —o 0 R,
-CH2CHZ2, -CH2CZ3, or j |j
R O
Q is -0-, -NH-, -S-, -CH20-, -CH2NH- , or -CH2S-; Z is -Cl, -Br, or -F; R, R', and R' ' are independently hydrogen, alkyl, cycloalkyl-(CH2)n-, aryl-(CH )n-, heteroaryl-(CH )n-, -CH2-(CH2)n-C02H, substituted cycloalkyl-(CH2)n-, substituted alkyl, substituted aryl-(CH2)n-, or substituted heteroaryl-(CH2)n-; and each n is independently 0 to 5, or the pharmaceutically acceptable salts, amides, esters, or prodrugs thereof.
The term "alkyl" means a straight or branched chain hydrocarbon. Preferably, the alkyl group has from 1 to 10 carbon atoms. More preferably, the alkyl group has from 1 to 6 carbon atoms. Examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, and tert-butyl. The term "cycloalkyl" means a cyclic hydrocarbon, which can be saturated or unsaturated. Preferably, the cycloalkyl group has from 3 to 10 carbon atoms. More preferably, the cycloalkyl group has from 3 to 6 carbon atoms. Examples of cycloalkyl groups include, but are not limited to, cyclopropane, cyclobutane, cyclopentane, cyclopentene, cyclohexane, cyclohexene, cycloheptane, cyclooctane, and adamantane.
The term "aryl" means a cyclic aromatic hydrocarbon. Examples of aryl groups include, but are not limited to, phenyl and naphthyl. The term "heteroaryl" means a cyclic aromatic hydrocarbon that contains one or more heteroatom. Examples of heteroatoms include, but are not limited to, oxygen, nitrogen, and sulfur. Examples of heteroaryl groups include, but are not limited to, pyridinyl, furanyl, thiophenyl, and pyrrolyl.
The terms substituted aryl, substituted phenyl, substituted cycloalkyl, substituted heteroaryl, or substituted alkyl mean an aryl, phenyl, cycloalkyl, heteroaryl, or alkyl group that has one or more substituent. Examples of substituents include alkyl, alkoxy, (such as methoxy, ethoxy, or tert-butoxy) , halogen, -N02, -OCH2CONH2, -OCH2C02H, -SO3H, -CONH2, -S02NH2, or -CH2OH, and the like. The symbol "-" means a bond. Examples of proteins that contain an SH2 domain include, but are not limited to, Src, Fyn, Lck, Yes, Blk, Lyn, Fgr, Hck, Yrk, and Abl. Preferably, the protein is Src, and most preferably the protein is pp60c-src. The term "cognate phosphorylated protein" means a protein to which the SH2 domain of a protein containing an SH2 domain binds or is associated. Examples of cognate phosphorylated proteins include, but are not limited to, PDGF receptor protein, EGF receptor protein, HER2/Neu receptor protein (an oncogene form of EGF receptor protein), fibroblast growth factor receptor protein, focal adhesion kinase protein, pl30 protein, and p68 protein.
The compounds and pharmaceutically acceptable compositions that contain the compounds can be administered to humans and animals either orally, rectally, parenterally (intravenous, by intramuscularly or subcutaneously) , intracisternally, intravaginally, intraperitoneally, intravesically, locally (powders, ointments or drops), or as a buccal or nasal spray. Compositions suitable for parenteral injection may comprise physiologically acceptable sterile aqueous or nonaqueous solutions, dispersions, suspensions, or emulsions and sterile powders for reconstitution into sterile injectable solutions or dispersions. Examples of suitable aqueous and nonaqueous carriers, diluents, solvents or vehicles include water, ethanol, polyols (propyleneglycol, polyethyleneglycol, glycerol, and the like), suitable mixtures thereof, vegetable oils (such as olive oil), and injectable organic esters such as ethyl oleate. Proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants.
These compositions may also contain adjuvants such as preserving, wetting, emulsifying, and dispensing agents. Prevention of the action of microorganisms can be ensured by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, and the like. It may also be desirable to include isotonic agents, for example, sugars, sodium chloride, and the like. Prolonged absorption of the injectable pharmaceutical form can be brought about by the use of agents delaying absorption, for example, aluminum monostearate and gelatin. Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound is admixed with at least one inert customary excipient (or carrier) such as sodium citrate or dicalcium phosphate or (a) fillers or extenders, as for example, starches, lactose, sucrose, glucose, mannitol, and silicic acid, (b) binders, as for example, carboxymethylcellulose, alignates, gelatin, polyvinylpyrrolidone, sucrose and acacia, (c) humectants, as for example, glycerol,
(d) disintegrating agents, as for example, agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates and sodium carbonate,
(e) solution retarders, as for example, paraffin,
(f) absorption accelerators, as for example, quaternary ammonium compounds, (g) wetting agents, as for example, cetyl alcohol and glycerol monostearate,
(h) adsorbents, as for example, kaolin and bentonite, and (i) lubricants, as for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets, and pills, the dosage forms may also comprise buffering agents.
Solid compositions of a similar type may also be employed as fillers in soft- and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethyleneglycols, and the like.
Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells, such as enteric coatings and others well known in the art. They may contain opacifying agents, and can also be of such composition that they release the active compound or compounds in a certain part of the intestinal tract in a delayed manner. Examples of embedding compositions which can be used are polymeric substances and waxes. The active compounds can also be in micro-encapsulated form, if appropriate, with one or more of the above-mentioned excipients.
Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs. In addition to the active compounds, the liquid dosage forms may contain inert diluents commonly used in the art, such as water or other solvents, solubilizing agents and emulsifiers, as for example, ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propyleneglycol, 1,3-butyleneglycol, dimethylformamide, oils, in particular, cottonseed oil, groundnut oil, corn germ oil, olive oil, castor oil and sesame oil, glycerol, tetrahydrofurfuryl alcohol, polyethyleneglycols, and fatty acid esters of sorbitan or mixtures of these substances, and the like.
Besides such inert diluents, the composition can also include adjuvants, such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.
Suspensions, in addition to the active compounds, may contain suspending agents, as for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, or mixtures of these substances, and the like.
Compositions for rectal administrations are preferably suppositories which can be prepared by mixing the compounds of the present invention with suitable non-irritating excipients or carriers such as cocoa butter, polyethyleneglycol or a suppository wax, which are solid at ordinary temperatures but liquid at body temperature and therefore, melt in the rectum or vaginal cavity and release the active component. Dosage forms for topical administration of a compound of this invention include ointments, powders, sprays and inhalants. The active component is admixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers or propellants as may be required. Ophthalmic formulations, eye ointments, powders, and solutions are also contemplated as being within the scope of this invention. The term "pharmaceutically acceptable salts, esters, amides, and prodrugs" as used herein refers to those carboxylate salts, amino acid addition salts, esters, amides and prodrugs of the compounds of the present invention which are, within the scope of sound medical judgement, suitable for use in contact with the tissues of patients without undue toxicity, irritation, allergic response, and the like, commensurate with a reasonable benefit/risk ratio, and effective for their intended use, as well as the zwitterionic forms, where possible, of the compounds of the invention. The term "salts" refers to the relatively non-toxic, inorganic and organic acid addition salts of compounds of the present invention. These salts can be prepared in situ during the final isolation and purification of the compounds or by separately reacting the purified compound in its free base form with a suitable organic or inorganic acid and isolating the salt thus formed. Representative salts include the hydrobro ide, hydrochloride, sulfate, bisulfate, nitrate, acetate, oxalate, valerate, oleate, palmitate, stearate, laurate, borate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, naphthylate mesylate, glucoheptonate, lactiobionate and laurylsulphonate salts, and the like. These may include cations based on the alkali and alkaline earth metals, such as sodium, lithium, potassium, calcium, magnesium, and the like, as well as nontoxic ammonium, quaternary ammonium and amine cations including, but not limited to ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine, and the like. (See, for example, Berge S.M., et al. , "Pharmaceutical Salts," J. Pharm. Sci. , 1977;66:1-19 which is incorporated herein by reference. ) Examples of pharmaceutically acceptable, non-toxic esters of the compounds of this invention include C-^-Cg alkyl esters wherein the alkyl group is a straight or branched chain. Acceptable esters also include C5-C7 cycloalkyl esters as well as arylalkyl esters such as, but not limited to benzyl. C-^-C^j alkyl esters are preferred. Esters of the compounds of the present invention may be prepared according to conventional methods. Particularly preferred esters are phosphoesters. Examples of suitable phosphoesters include, but are not limited to -P03RcRd,
-CF2P03RcRd, and -CH2P03RcRd, where Rc and Rd are as defined above. (See, for example, Jones R.J. and Bischofberger N., "Minireview: nucleotide prodrugs", Antiviral Research, 1995;27:1-17, which is incorporated herein by reference.
Examples of pharmaceutically acceptable, non-toxic amides of the compounds of this invention include amides derived from ammonia, primary C-^-Cg alkyl amines and secondary Cj-C dialkyl amines wherein the alkyl groups are straight or branched chain. In the case of secondary amines, the amine may also be in the form of a 5- or 6-membered heterocycle containing one nitrogen atom. Amides derived from ammonia, C-^- ^ alkyl primary amides and C _C2 dialkyl secondary amides are preferred. Amides of the compounds of the invention may be prepared according to conventional methods. The term "prodrug" refers to compounds that are rapidly transformed in vivo to yield the parent compound of the above formulas, for example, by hydrolysis in blood. A thorough discussion is provided in T. Higuchi and V. Stella, "Pro-drugs as Novel
Delivery Systems," Vol. 14 of the A.C.S. Symposium Series, and in Bioreversible Carriers in Druσ Desiσn, ed. Edward B. Roche, American Pharmaceutical Association and Pergamon Press, 1987, both of which are incorporated herein by reference.
The compounds of the present invention can be administered to a patient at dosage levels in the range of about 1 to about 7,000 mg/day. For a normal human adult having a body weight of about 70 kg, a dosage in the range of about 1 to about 100 mg/kg of body weight per day is sufficient. The specific dosage used, however, can vary. For example, the dosage can depend on a number of factors including the requirements of the patient, the severity of the condition being treated, and the pharmacological activity of the compound being used. The determination of optimum dosages for a particular patient is well known to those skilled in the art.
The compounds of the present invention can exist in different stereoisometric forms by virtue of the presence of asymmetric centers in the compounds. It is contemplated that all stereoisometric forms of the compounds as well as mixture thereof, including racemic mixtures, form part of this invention. In addition, the compounds of the present invention can exist in unsolvated as well as solvated forms with pharmaceutically acceptable solvents such as water, ethanol, and the like. In general, the solvated forms are considered equivalent to the unsolvated forms for the purposes of the present invention. The examples presented below are intended to illustrate particular embodiments of the invention and are not intended to limit the specification or the claims in any manner.
EXAMPLES The following abbreviations may be used in the present application:
Abu α-Aminobutyric acid
Ac Acetyl
Ala Alanine
Arg Arginine
Asn Asparginine Asp Aspartic acid
Boc Tertiary butyloxycarbonyl
Bn Benzyl
Cbz Benzyloxycarbonyl
Cys Cysteine CF3S02H Trifluoromethanesulfonic acid
DCC N,N' -Dicyclohexylcarbodiimide
DCM Dichloromethane
DIC N,N' -Diisopropylcarbodiimide
DIEA N,N' -Diisopropylethylamine DMAP 4-Dimethylaminopyridine
DMF N,N' -Dimethylformamide
EDCI N-Ethyl-N' -Dimethylaminopropyl- carbodiimide EtOAc Ethyl acetate
Et20 Diethyl ether
Fmoc 9-Fluorenylmethyloxycarbonyl
Gin Glutamine
Glu Glutamic acid
Gly Glycine
HCl Hydrochloric acid
His Histidine
HOAc Acetic acid
HOBT 1-Hydroxybenzotriazole
He Isoleucine iprOH Isopropanol
KOH Potassium hydroxide
Leu Leucine
Lys Lysine
MeCN Acetonitrile
MeOH Methanol
Met Methionine
NHOS N-Hydroxysuccinimide
NMP N-Methylpyrrolidone
Phe Phenylalanine
Pro Proline rt Retention time
Ser Serine
TFA Trifluoroacetic acid
THF Tetrahydrofuran
Thr Threonine Trp Tryptophan
Tyr Tyrosine
Val Valine
EXAMPLE 1 rS- ( R* .R* ) λ -4- f2-Acetvlamino-3- ( -phosphonooxy-phenyl ) propionvlaminol -4- \ (3-cyclohexyl -propyl^-methyl- carbamoyll-butyric acid or Ac- (O-phosphono)-L-Tyr-L-Glu-Nfmethyl^ (3-cyclohexyl- propyl)
Figure imgf000028_0001
Step 1 : Fmoc-L-Glu t 0tBu ^ -N tmethyl 3 -cyclohexyl - propyl )
To methyl (3-cyclohexylpropyl) amine hydrochloride (5.0 mmol, 960 mg) in tetrahydrofuran (20 mL) was added Fmoc-L-Glu(OtBu) (5.5 mmol, 2.34 g) followed by sequential addition of 1-hydroxybenzotriazole (6.25 mmol, 845 mg), N-methylmorpholine (12.5 mmol, 1.37 mL) , and 1-(3-dimethylaminopropyl) -3-ethyl- carbodiimide hydrochloride (6.25 mmol, 1.2 g) . After stirring 16 hours at room temperature, diethyl ether was added, and the remaining residue was dissolved in water. The mixture was separated, washed with 10% sulfuric acid, water, then saturated sodium bicarbonate, and then brine to provide product as a colorless foam (2.59 g, 92%). 1H NMR (CDC13^ 300 MHz): δ 0.87 (m, 2H), 1.17 (m, 6H) , 1.46 (s, 9H) , 1.50-1.80 (m, 8H), 2.05 (m, IH) , 2.33 (m, 2H) , 2.95-3.11 (s, 3H, rotational isomers), 3.18-3.58 (m, 2H), 4.21 (t, IH) , 4.36 (m, 2H) , 4.70 (m, IH), 5.80 (dd, IH), 7.27-7.45 (m, 4H) , 7.60 (m, 2H) , 7.77 (d, 2H); IR (CHC13): 1719, 1642 cm"1; Mass Spectrum (Chemical Ionization, 1% NH3 in CH4) m/z 563(M+H).
Step 2; Ac -L-Tvr-L-Glu t OtBm -NfmethyH
(3-cyclohexylpropyl ) To Fmoc-L-Glu(OtBu)-N(methyl) (3-cyclohexylpropyl)
(2.0 mmol, 1.12 g) in dichloromethane (20 L) was added piperidine (4 L) . After 20 minutes, toluene (20 mL) was added, and the solvent was removed under reduced pressure. Toluene (20 mL) was again added, and the solvent was evaporated. The resulting residue was dissolved in tetrahydrofuran (15 mL) and coupled with Ac-L-Tyr (2.2 mmol, 491 mg) in the manner described above to give a solid residue upon work-up. Chromatography of the residue (3:7, tetrahydrofuran/ dichloromethane) gave product as a colorless foam (880 mg, 81%). 1H NMR (DMSO, 300 MHz): 6 0.85 (m, 2H), 1.03-1.30 (m, 6H), 1.40 (s, 9H) , 1.40-1.90 ( , 10H), 1.75 (s, 3H) , 2.20 (t, 3H) , 2.60 (dd, IH) , 2.78-2.95 (s, 3H, rotational isomers), 3.10-3.30 (m, 2H), 4.44 (m, IH) , 4.70 (m, IH), 6.61 (d, 2H) , 7.00 (d, 2H), 7.96 (d, IH), 8.01 (t, 3H) , 9.15 (s, IH); IR (CHCI3): 1720, 1639 cm"1; Mass Spectrum (Chemical Ionization, 1% NH3 in CH4) m/z 546 (M+H) .
Step 3: Ac-(O-phosphono^ -L-Tyr-L-Glu-N tmethyl ^ (3-cyclohexylpropyl )
To Ac-L-Tyr-L-Glu(OtBu) -N(methyl) (3-cyclohexyl- propyl) (0.40 mmol, 218 mg) in dichloromethane (5 mL) was added tetrazole (0.80 mmol, 56 mg) and di-tert- butyl diethylphosphoramidate (0.60 mmol, 0.17 mL) . After 3 hours, thin layer analysis indicated the reaction was complete and tert-butylhydroperoxide (2.4 mmol, 0.22 mL) was added. After 2 hours, ethyl acetate was added, and the organic phase was washed with 10% sulfuric acid, water, 2% sodium hydroxide, and then saturated sodium chloride. The solvent was removed under reduced pressure to give an oil (432 mg) . This oil was treated with trifluoroacetic acid (15 mL) and water (1 mL) for 1 hour. The solvent was removed under reduced pressure, and the residue was precipitated with diethyl ether to give a colorless solid. Preparative HPLC of crude product (75 mg) employing a Vydac C18 (22 x 250 mm) column eluting with a gradient of 0% to 30% acetonitrile containing 0.1% trifluoroacetic acid and water containing 0.1% trifluoroacetic acid (TFA) provided pure product after lyophilization (31 mg) . HPLC 100%, rt - 17.3 minutes, C18 (analytical column, Vydac, 4.6 x 250 mm), eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 569.5 (M-H) .
EXAMPLE 2 4- f (S )-2-Acetylamino-3-f4-phosphonooxy-phenyl )- (S ) propionylaminol-4-f2-cyclohexyl- fS^ -1-methyl- ethvlcarbamoγl^-butvric acid or Ac-fO-phosphono^-L-Tyr-L-Glu-NHr- ( S \-2-cyclohexyl-
1-me hylethyl)
H2O3P
Figure imgf000030_0001
The title compound was synthesized in a manner similar to that described for Example 1. Product was obtained as a colorless solid (160 mg) . HPLC 100%, rt = 16.2 minutes, C18, eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 555.4 (M-H) .
EXAMPLE 3
4- r (S ) -2-Acetvlamino-3- (4-phosphonooxy-phenyl - (S ) - propionyl-amino1 -4 - t 2-cyclohexyl- f S ^ -l-methyl -ethyl ^ - methyl-carbamoyl)-butyric acid or Ac-f-Q-phosphono^-L-Tyr-L-Glu-Nfmethyl ( ( S ) - 2-cyclohexyl-1-methylethy1 )
Figure imgf000031_0001
The title compound was synthesized in a manner similar to that described for Example 1. Product was obtained as a colorless solid (117 mg) . HPLC 93%, rt = 17.5 minutes, C18, eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 569.5 (M-H). EXAMPLE 4 4-f CRS^ -2-Acetylamino-3- f4- fdifluoro-phosphono-methyl phenyl1 - (S ) -propionylaminol -4- r (3-cyclohexyl-propyl ) - methvl-carbamoy11 -butyric acid or Ac- (4- (difluorophosphonomethyl ) -. -D/L-Phe-L-Glu- Nfmethyls f3-cyπlohexγlpropvl^
Figure imgf000032_0001
Step 1; gpc- r4-(diethoxyphosphonyl)- difluoromethyll-D/L-Phe Benzyl ester
Boc- [4- (diethoxyphosphonyl) -difluoromethyl] - D/L-Phe Benzyl ester can be prepared in accordance with methods well known to those skilled in the art. (See, for example, Burke, et al . , J. Org. Chem. , 1993;58(6) :1336-1340. )
Step 2: Ac- f4- tdiethoxyphosphonyl^- difluoromethyll -D/L-Phe
Ac- [4- (diethoxyphosphonyl)difluoromethyl] -D/L-Phe was prepared in a manner similar to that described for Example 5 (Step 1).
Step 3: Ac- (4- (difluorophosphonomethyl ) ) -D/L-Phe- L-Glu-N(methyl (3-cvclohexvlpropvl>
The title compound was then synthesized in a manner similar with the substitution of Example 1, Ac- [4- (diethoxyphosphonyl) -difluoromethyl] -D/L-Phe for Ac-L-Tyr. The crude peptide was deprotected with trifluoroacetic acid/trimethysilyl triflate/ ethanedithiol/m-cresol/dimethylsulfide (4/1/0.1/ 0.003/1) for 2 hours at room temperature. Careful addition of water then ether quenched the reaction. Preparative HPLC, as previously described, of the aqueous layer provided product as a colorless solid after lyophilization (29 mg) . HPLC 93%, rt = 15.1 minutes, C18, eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 603.4 (M-H).
EXAMPLE 5 rS- t R*,R* )1 -4-(2-Acetvlamino-3- r4- <difluoro-phosphono- methvl^-phenvll -pro ionvlamino)-4- r (3-cyclohexyl- propyl -methvl-carbamovl1 -butyric acid or Aς-(4-(di£luorophosphonomethyl))- -Phe- -Glu-Nfmethyl)
( -cyclohexvlpropvl
Figure imgf000033_0001
Step 1 ; Ac- r 4 - ( diethoxyphosphonyl ^ - difluoromethy11- -Phe
Boc- [4-(diethoxyphosphonyl) -difluoromethyl] -L-Phe Benzyl ester can be prepared in accordance with methods well known to those skilled in the art. (See, for example, Smythe and Burke, Tett. Lett.. 1994;35(4) :551- 554. ) Boc- [4-(diethoxyphosphonyl)-difluoromethyl] -L- Phe Benzyl ester(1.75 mmol, 950 mg) was deprotected with 20 mL trifluoroacetic acid:dichloromethane (1:1) for 5 hours at 0°C. The reaction was diluted with 200 mL of ethyl acetate and washed with saturated sodium bicarbonate then saturated sodium chloride, dried with sodium sulfate, filtered, and concentrated under reduced pressure to yield 795 mg of a colorless oil. The crude oil was treated with acetic anhydride (10 mmol, 944 μL) and pyridine (15 mmol, 1.21 mL) in 20 mL dichloromethane for 4 hours at room temperature then 2 days at 4°C. The reaction was diluted with 400 mL of ethyl acetate and washed with saturated sodium bicarbonate, 5% hydrochloric acid, saturated sodium bicarbonate, then saturated sodium chloride, dried with sodium sulfate, filtered, and concentrated under reduced pressure to yield 1.2 g of a pale yellow oil. Chromatography of the residue (20:1, dichloromethane:methanol) gave 719 mg (85%) of the product as a colorless oil. Removal of the benzyl ester with hydrogen and palladium on carbon (20%) in ethanol gave, after filtration and concentrating under reduced pressure, 564 mg (99%) of a solid foam. 1H NMR (DMSO, 400 MHz): δ 1.20 (t, 6H) , 1.76 (s, 3H) , 2.90 (dd, IH), 3.12 (dd, IH), 4.07 (m, 4H), 4.44 (m, IH) , 7.39 (d, 2H), 7.46 (d, 2H) , 8.22 (d, IH) ; Mass Spectrum (Chemical Ionization, 1% NH3 in CH4) m/z 394 (M+H) . step 2: &c-(4-(Difluorpp osphonomethy ) )~L-Phe- - Glu-N(methyl> (3-cγclohexvlpropvl^
The title compound was synthesized in a manner similar to Example 4 only Ac- [4-(diethoxyphosphonyl) difluoromethyl] -L-Phe was coupled rather than the D/L mixture. The purified peptide (30 mg) was deprotected with 1 M trimethysilyl triflate and 2 M dimethylsulfide in trifluoroacetic acid (3 mL) for 16 hours at room temperature. Water was added to quench excess trimethysilyl triflate, and the resulting solution was concentrated at reduced pressure to remove volatiles. The remaining solution was diluted with trifluoroacetic acid and water and purified by preparative HPLC, as previously described, to provide the product as a colorless solid after lyophilization (22 mg) . HPLC 100%, rt = 16.6 minutes, C18, eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 603 (M-H) .
EXAMPLE 6 rs- (R*,R 1 -4- r (3-Cyclohexyl-propyl^ -methyls - carbamovll -4- \ 2 - \ fmorpholine-4-carbonyl^ -amino1 i phosphonooxy-phenvl^ -propionylaminol -butyric acid or 4-Morpholinecarbonvl- (O-phosphono^ -L-Tγr-L-Glu- N (methyl 3-cyclohexylpropyl1
Figure imgf000035_0001
Step 1: 4 -Morpholinecarbonγl - L- Tyr t Bzl -OBzl
To H-Tyr(Bzl) -Obzl -p-tosylate (7.5 mmol, 4.0 g) in dichloromethane (50 mL) at 0°C was added N-methylpiperidine (7.5 mmol, 0.75 g) followed by dropwise addition of 4-morpholinecarbonyl chloride (7.5 mmol, 1.13 g) . After stirring 12 hours at room temperature, the mixture was washed sequentially with 10% sulfuric acid, water, saturated sodium bicarbonate and brine to provide product as an oil (3.81 g, 99%). Step 2; 4-Morpholinecarbonγl-L-Tyr-OH
A mixture of 4-Morpholinecarbonyl-L-Tyr(Bzl)-OBzl (8.0 mmol, 3.81 g), methanol (50 mL) , and 10% palladium on carbon (0.50 g) was stirred under a hydrogen atmosphere for 20 hours. The mixture was filtered over Celite, and the solvent was removed at reduced pressure to give (2.35 g, 97%) of the product as an oil.
Step 3; 4 -Morpholinecarbonγl -L-Tyr-L-Glu t OtBm -
N(methyls (3-cvclohexvlpropvl )
To Fmoc-LGlu(OtBu)- (methyl) (3-cyclohexylpropyl) (2.0 mmol, 1.12 g) in dichloromethane (20 mL) was added piperidine (4 mL) . After 20 minutes toluene (20 mL) was added, and the solvent was removed under reduced pressure. Toluene (20 mL) was again added, and the solvent was evaporated. The resulting residue was dissolved in Dimethylformamide (20 mL) and coupled with the 4-Morpholine-carbonyl-L-Tyr-OH (2.0 mmol, 600 mg) in a manner similar to that described for Example 1 to give a solid residue upon work-up. Chromatography of the residue (2:8, tetrahydrofuran/dichloromethane) gave product as a colorless foam (700 mg, 63%) .
Step 4 : 4-Morpholinecarbonyl-(Q-phosphono -L-Tyr-
L-Glu-Nfmethvl (3-cvclohexvlpropvl^
The reaction was carried out in a manner similar to that described for Example 1 (Step 3). The product was obtained as a white solid (83 mg) . HPLC 89%, rt = 17.7 minutes, C18, eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 639.4. EXAMPLE 7 r s- tR* , R* n - 4 - r t 3 -Cyclohexyl -propyl ^ -methyl-carbamovl 1 4- r2- <3-methyl-ureido )-3- <4-phosphonooxv-phenvl )- propionylaminol-butyric acid or
N- rfMethylamino^carbonyl) - rθ-phosphono^ -L-Tγr-L-Glu- N(methyl ) (3-cyclohexylpropy1 )
Figure imgf000037_0001
Step 1; N- r (Methylamino )carbonyl) -L-Tγr(BzH -OBzl
To H-Tyr(Bzl)-OBzl -p-tosylate (11.2 mmol, 6.0 g) in tetrahydrofuran (50 L) at 0°C was added N-methylpiperidine (3.60 mL) followed by dropwise addition of methyl isocyanate (14.6 mmol, 0.85 g) in tetrahydrofuran (5 mL) . After stirring 2 hours at room temperature, the mixture was evaporated to dryness under reduced pressure. Chromatography of the residue (1:4, ethyl acetate:hexane) provided the product as a white foam (4.72 g, 99%).
Step 2: N- r tMethγlamino ^carbonyl1 -L-Tyr-OH A mixture of N-methylcarbonyl-L-Tyr(Bzl)-OBzl (4.8 mmol, 2.0 g), methanol (50 mL), and 10% palladium on carbon (0.50 g) was stirred under a hydrogen atmosphere for 20 hours. The mixture was filtered over Celite, and the solvent was removed at the reduced pressure to give (1.27 g, 97%) of the product as an oil. Step 3; N- r tMethvlamino ^carbonyl 1 -L-Tγr-L-
GlufOtBu)-NCmethvl) (3-cvclohexvlpropy1 \
To Fmoc-L-Glu(OtBu)-N(methyl) (3-cyclohexylpropyl) (5.3 mmol, 2.98 g) in dichloromethane (20 mL) was added piperidine (4 mL) . After 20 minutes toluene (20 mL) was added, and the solvent was removed under reduced pressure. Toluene (20 mL) was again added, and the solvent was evaporated. The resulting residue was dissolved in dimethylformamide (20 mL) and coupled with the N-methylcarbonyl-L-Tyr-OH (5.3 mmol, 1.27 g) in a manner similar to that described for Example 1 to give a solid residue upon work-up. Chromatography of the residue (2:8, tetrahydrofuran/dichloromethane) gave product as a colorless foam (2.0 g, 67%). Step 4; N- Γ (Methvlamino^carbonyl) -(0-phosphono^-
L-Tvr-L-Glu-Ntmethγl > (3-cvclohexvlpropvl^
The reaction was carried out in a manner similar to that described for Example 1 (Step 3). Product was obtained as a white solid (67 mg) . HPLC 97%, rt = 18.4 minutes, C18, eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 583.5 (M-H).
EXAMPLE 8 rs-(R*,R* -Phosphoric acid mono- ι~4-f2-acetylamino-2- propylcarbamoyl)-ethvl)-phenyl) ester or Ac- f O-phosphono ^ -L-Tyr-L-Abu-NtmethyH t 3 -cγclohexγl- propy )
Figure imgf000039_0001
The title compound was synthesized in a manner similar to that described for Example 1. The product was obtained as a colorless solid (78 mg) . HPLC 100%, rt = 19.2 minutes, C18, eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 524.4 (M-H).
EXAMPLE 9 rs- t R* . R l -Phosphoric acid mono- - (2-acetvlamino-2- f1- r t3-cyclohexyl-propyl ) -methyl-carbamoyl) - pentvlcarbamoyl) -ethyls -phenyl) ester or Ac- fO-phosphono^-L-Tγr-L-Nle-N(methyls (3-cvclohexvl- propyl)
Figure imgf000039_0002
The title compound was synthesized in a manner similar to that described for Example 1. The product was obtained as a colorless solid (92 mg) . HPLC 100%, rt = 21.5 minutes, C18, eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 552.4 (M-H).
EXAMPLE 10 r s- tR* , R ) - f r4-(2-Acetylamino-2-fl-r(3-cγclohexyl- propyl^ -methvl-carbamovl) -propvlcarbamoyl)-ethyls - phenyl) -difluoro-methvl)-phosphonic acid or Ac- (4- tdifluorophosphonomethyln-L-Phe-L- bu-N(methyl (3-cyclohexylpropyl)
Figure imgf000040_0001
Step 1: Boc-L-Abu-N- (methyl 3-cγclohexγlpropγH Boc-L-Abu- (methyl) (3-cyclohexylpropyl) was synthesized in a manner similar to that described for Example 1 (Step 1) to yield a colorless oil. XH NMR (CDC13, 400 MHZ) : δ 0.87 (m, 2H) , 0.93 (t, 3H), 1.17 (m, 6H), 1.43 (s, 9H) , 1.50-1.80 (m, 7H) , 2.92 + 3.03 (s, 3H, rotational isomers), 3.23-3.44 (m, 2H) , 4.54 (m, IH), 5.40 (2d, IH, rotational isomers). Step 2: Ac-(4-(difluorophosphonomethyln-L-Phe-L- Abu-N(methyl ) (3-cγclohexvlpropyl )
Ac-(4-(difluorophosphonomethyl) )-L-Phe-L-Abu-N- (methyl) (3-cyclohexylpropyl) was synthesized in a manner similar to that in Example 5 (Step 2) only Boc- L-Abu-N-(methyl) (3-cyclohexylpropyl) was used rather than Fmoc-L-Glu-N(methyl) (3-cyclohexylpropyl) , and was deprotected with TFA. The purified peptide (80 mg) was deprotected with 1 M trimethysilyl triflate and 2 M dimethylsulfide in trifluoroacetic acid (5 mL) for
3 hours at 0°C and 1 hour at room temperature. Water was added to quench excess trimethysilyl triflate, and the resulting solution was concentrated under reduced pressure to remove volatiles. The remaining solution was diluted with trifluoroacetic acid and water and purified by preparative HPLC, as previously described, to provided the product as a colorless solid after lyophilization (51 mg) . HPLC 100%, rt = 18.0 minutes, C18, eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 558 (M-H).
EXAMPLE 11
4 - r ( S ^ - 2 -Acetvlamino- 3 - ( 4 -phosphonooxγ-phenvH - S ^ - propionvlamino) - 5 -OXO- 5 - ( S ^ - 2 -phenethyl -pγrrolidin-l- vH -pentanoic acid
Figure imgf000042_0001
The title compound was synthesized in a manner similar to that described for Example 1. The product was obtained as a colorless solid (51 mg) . HPLC 100%, rt = 15.7 minutes, C18, eluting with a gradient of 10% to 76% acetonitrile containing 0.1% TFA, and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 595.5 (M-H).
EXAMPLE 12 4- r (S ) -2-Acetylamino-3- ( -phosphonooxy-phenyl ) - ( S ) - propionylamino)-5-l~2-((S^-2-cyclohexyl-ethyl ) - pγrrolidin-1-yl) -5-oxo-pentanoic acid
Figure imgf000042_0002
The title compound was synthesized in a manner similar to that described for Example 1. The product was obtained as a colorless solid (51 mg) . HPLC 100%, rt = 13.3 minutes, C18 , eluting with a gradient of 10% to 76% acetonitrile containing 0.1% TFA, and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 589.4 (M-H).
EXAMPLE 13 fS- (R* ,R*) 1 -4- r2-Acetylamino-3- (4-phosphonooxv-phenyl ) - propionvlamino)-4- \2- (5, 6 , 7 , 8-tetrahydro-naphthalen- 1-vl^-ethvlcarbamoγll -butyric acid
Figure imgf000043_0001
The title compound was synthesized in a manner similar to that described for Example 1. The product was obtained as a colorless solid (178 mg) . HPLC 93%, rt = 15.6 minutes, C18, eluting with a gradient of 10% to 76% acetonitrile containing 0.1% TFA, and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 588.3 (M-H). EX MPLE 14
Figure imgf000044_0001
propionvlamino) -4-(3-phenγl-propγlcarbamoγl^-butyric acid or Ac-(Q-phosphono^L-Tγr-L-Glu-NH(3-phenγlpropγl^
The title compound was synthesized in a manner similar to that described for Example 1. The product was obtained as a colorless powder (52 mg) . HPLC 100%, rt = 14.0 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 548.3 (M-H).
EXAMPLE 15 fS-(R*,R )-4- r2-Acetγlamino-3-(4-phosphonooxv-phenγl propionylamino) -4- T (naphthalen-1-ylmethγl^-carbamoyl) - butvric acid
Figure imgf000045_0001
The title compound was synthesized in a manner similar to that described for Example 1. The product was obtained as a colorless powder (35 mg) . HPLC 100%, rt «= 14.6 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 571.5 (M-H).
EXAMPLE 16 fS- (R* .R* -4- r2-Acetylamino-3- (4-phosphonooxv-phenyl ) - propionylamino) -4-phenethylcarbamoyl-butyric acid or Ac-(0-phosphonoϊL-Tγr-L-Glu-NH(2-phenγlethvl^
Figure imgf000045_0002
The title compound was synthesized in a manner similar to that described for Example 1. The product was obtained as a colorless powder (27 mg) . HPLC 100%, rt = 12.5 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 534.5 (M-H).
EXAMPLE 17 4- r (S )-2-Acetylamino-3-(4-phosphonooxy-phenyl ) - (S ) - propionvlamino) -4- fmethvl-( (S^-1-methyl-2-phenyl- ethyl )-carbamoyl) -butyric acid or Ac-(Q-phosphono^-L-Tyr-L-Glu-N(methyl^ ( (S^-1-methvl-
2-phenylethy )
Figure imgf000046_0001
The title compound was synthesized in a manner similar to that described for Example 1. The product was obtained as a colorless solid (176 mg) . HPLC 91%, rt = 13.8 minutes, C18, eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 563.4 (M-H). EXAMPLE 18 4- r (S) -2-Acetylamino-3- (4-phosphonooxy-phenvl) - (S ) propionylamino) -4- ( (SI -1-methyl-2-phenyl- ethylcarbamoyll -butyric acid or
Ac- (0-phosphono) -L-Tyr-L-Glu-NH( (SI -l-methyl-2- phenvlethyll
Figure imgf000047_0001
The title compound was synthesized in a manner similar to that described for Example 1. The product was obtained as a colorless solid (251 mg) . HPLC 100%, rt = 13.6 minutes, C18, eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 549.4 (M-H).
EXAMPLE 19 TS- (R* ,R* -4- r2-Acetvlamino-3- (4-phosphonooxy-phenyl ) - propionylamino) -4- rmethyl- (3-phenyl-propyl ) -carbamoyl) - butvric acid or Ac- (O-phosphonol -L-Tγr-L-Glu-N(methyl> (3-phenγlpropyl1
The title compound was synthesized in a manner similar to that described for Example 1. The product was obtained as a colorless solid (108 mg) . HPLC 85%, rt = 14.5 minutes, C18, eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 562.5 (M-H).
EXAMPLE 20 rs-(R*,R* -Phosphoric acid mono- (4- f2-acetylamino-2- r2- (4-hydroxy-phenyl1 -1- (3-phenyl-propylcarbamovl1 - ethyl-carbamoyl) -ethyl)-phenyl ) ester
Figure imgf000048_0002
The title compound was synthesized in a manner similar to that described for Example i. The product was obtained as a colorless powder (110 mg) . HPLC 100%, rt = 15.6 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 584.3 (M-H).
EXAMPLE 21 TS-(R* , R* ) ) -4- r2-Acetylamino-3- (4-phosphonooxy-phenyl1 propionylamino) -4- f2- (2-methoxy-pheny11- ethylcarbamovl) -butyric acid or
Ac-(0-phosphonol-L-Tyr-L-Glu-NHf2- (2-methoxyphenyl1- ethyl)
Figure imgf000049_0001
The title compound was synthesized in a manner similar to that described for Example 1. The product was obtained as a colorless powder (62 mg) . HPLC 98%, rt = 14.6 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 579.4 (M-H). EXAMPLE 22 rS- (R* ,R*11 -4- r2-Acetylamino-3- (4-phosphonooxy-phenyl ) propionylamino) -4- (2-p-tolyl-ethylcarbamoyll -butyric acid or Ac- (0-phosphono 1 -L-Tvr-L-Glu-NH r 2 - t 4 -methylphenγl > - ethyll
Figure imgf000050_0001
The title compound was synthesized in a manner similar to that described for Example 1. The product was obtained as a colorless powder (109 mg) . HPLC 92%, rt = 13.7 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 548.4 (M-H).
EXAMPLE 23 rS- (R* ,R*1 ) -4- r2-Acetylamino-3- (4-phosphonooxy-phenyl1 - propionylamino) -4- f r2- (2-methoxy-phenyl1 -ethvl) -methyl• carbamoyl ) -butvric acid or
Ac-(0-phosphono1-L-Tyr-L-Glu-N(methyl1 f2-(2- methoxyphenvl1 -ethvl1
Figure imgf000051_0001
Step 1; N-(t-ButylQxycarbpny )-2-f2- methoxyphenyl1-ethylamine
To 2-methoxyphenylethylamine (163 mmol, 24 g) in tetrahydrofuran (150 mL) at 0°C was added triethylamine (180 mmol, 25 mL) followed by dropwise addition of di- t-butyloxycarbonyl dicarbonate (180 mmol, 39.15 g) in 75 mL of tetrahydrofuran. After 12 hours, the solvent was evaporated. The resulting residue was dissolved in ethyl acetate (150 mL) , washed with 10% sulfuric acid, then saturated sodium bicarbonate to provide the product as white solid (45 g, 99%).
Step 2: N-methvl-2-(2-methoxyphenyl1-ethylamine
To N-(t-Butyloxycarbonyl)-2-(2-methoxyphenyl) - ethylamine (182.7 mmol, 45.9 g) in dry tetrahydrofuran (200 mL) at 0°C was added, in portions, lithium aluminum hydride (219.2 mmol, 8.3 g) under N2. After stirring at 0°C for 1 hour, the ice-bath was removed and then the reaction heated to reflux for 24 hours, cooled to room temperature then to 0°C; the excess lithium aluminum hydride was destroyed very carefully with 20% potassium hydroxide solution (1.3 equivalent). After removing the salt by filtration, the filtrate was dried then evaporated to dryness to give product as a white solid (26.2 g, 87%).
Step 3 : Fmoc-L-Glu ( 0tBu 1 - N dnethγl H 2 - ( 2 - methoxyphenyl -ethyl) The compound was synthesized in a manner similar to that described for Example 1 (first step).
Step 4: Ac-L-Tyr-L-Glu(OtBu1-N(methyl 2-(2- methoxypheny11 -ethyl1
The compound was synthesized in a manner similar to that described for Example 1 (second step) .
Step 5; Ac- (0-phosphonol -L-Tγr-L-Glu-N(methγl1 \ 2 -
( 2-methoxyphenyl1 -ethyl1
The title compound was synthesized in a manner similar to that described for Example 1. The product was obtained as a white solid (400 mg) . HPLC 100%, rt = 13.2 minutes, C18, eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 578.4 (M-H).
EXAMPLE 24 rs-(R*,R*11 -4- r2-Acetylamino-3-(4-phosphonooxy-phenyl1 propionylamino) -4- T3- (3-carbamoylmethoxy-phenvll - propylcarbamoyl) -butyric acid or Ac- (O-phosphonol -L-Tyr-L-Glu-NHf3- r3- (0-acetamidol - phenyl]propyl)
Figure imgf000052_0001
Step 1: 3- (3-Hydroxyphenyl ) -propyl alcohol
To 3- (3-hydroxyphenyl) -propionic acid (60.2 mmol, 10.0 g) in dry tetrahydrofuran (200 mL) at 0°C was added, in portions, lithium aluminum hydride (72.2 mmol, 2.74 g) under N2. After stirring at 0°C for 1 hour, the ice-bath was removed and then the reaction heated to reflux for 24 hours, cooled to room temperature then to 0°C; the excess lithium aluminum hydride was destroyed very carefully with 20% potassium hydroxide solution (1.3 equivalent). After removing the salt by filtration, the filtrate was dried then evaporated to dryness to give product as a white solid (7.2 g, 78%) .
Step 2: 3- r3-(0-acetamido1 -phenyl) -propvl alcohol To 3- (3-hydroxyphenyl) -propyl alcohol (10.73 mmol,
1.63 g) in acetone (75 mL) was added 2-bromoacetamide (11.80 mmol, 1.62 g) and potassium carbonate (11.80 mmol, 1.63 g) . The reaction mixture was heated to reflux for 48 hours, cooled to room temperature, filtered, then the filtrate was concentrated to give product as white solid (2.20 g, 98%).
Step 3: 3- \ 3 - (O-acetamido1 -phenvl) -propvl- (O- methanesulfonyl1
To 3- [3- (O-acetamido) -phenyl] -propyl alcohol (11.80 mmol, 2.20 g) in dichloromethane (50 mL) at -10°C (ice/methanol) was added triethylamine (17.70 mmol, 2.5 mL) , then dropwise addition of methanesulfonyl chloride (12.40 mmol, 1 mL) . After stirring at -10°C for 1 hour, the reaction mixture was washed with water, 10% H2S04, saturated sodium bicarbonate, dried (MgS04), filtered, and concentrated to give product as an oil (2.98 g, 88%).
Step 4: 3- 3-(0-acetamido> -phenvl) -propvl azide
To 3- [3- (O-acetamido) -phenyl] -propyl- (0-methane- sulfonyl) (10.40 mmol, 2.98 g) in dimethylformamide (30 mL) was added sodium azide (13.6 mmol, 0.90 g) . After stirring at room temperature for 48 hours, the reaction mixture was quenched using water, then extracted with ether (4x100 mL) , washed with saturated sodium bicarbonate, dried (MgS04), filtered, then concentrated to give product as colorless oil (1.90 g, 78%) .
Step 5; 3- f3-(O-acetamido)-Phenvl) -propyl amine
3- [3-(O-acetamido) -phenyl] -propyl azide (8.10 mmol, 1.90 g) was catalytically reduced using 5% palladium on carbon (0.50 g) in tetrahydrofuran
(75 mL) . Hydrogenation was carried out on a Parr apparatus for 15 hours at 50 psi H2. After filtration, the solvent was removed under reduced pressure. The product was obtained as a white solid (1.28 g, 75%). Step 6: Fmoc-L-Glu(OtBu ) -NHf3- f3-(O-acetamido1 - pheny 1-propyl)
This compound was synthesized in a manner similar to that described for Example 1 (Step 1) .
Step 7: Ac-L-Tyr-L-Glu(OtBu1-NHf3- r3-(O- acetamido1-phenyl) ropyl)
This compound was synthesized in a manner similar to that described for Example 1 (Step 2).
Step 8: Ac- (O-phosphonol-L-Tyr-L-Glu-NHf3- \3-(0- acetamido1-phenyl)propvl) The title compound was synthesized in a manner similar to that described for Example 1. The product was obtained as a white solid (70 mg) . HPLC 93%, rt = 12.2 minutes, C18, eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 622.4 (M-H). EXAMPLE 25 rs- (R*,R*1 ) -4- r (2- r2-Acetylamino-3- (4-phosphonooxv phenvll -propionylamino) -propionyl) - (3-cvclohexvl- propyl ) -amino) -butyric acid or Ac-(0-phosphonol-L-Tvr-L-Ala-N(3-carboxvpropyl1
( -cyclohexylprppyl)
Figure imgf000055_0001
The title compound was synthesized in a manner similar to that described for Example 1. The product was obtained as a colorless powder (51 mg) . HPLC 100%, room temperature 19.9 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 582.3 (M-H).
EXAMPLE 26 rs-(R*,R*11-4-rf2-r2-Acetylamino-3-(4-phosphonooxy- phenvll -propionylamino) -propionyl) - (3-phenyl-propyl ) amino) -butyric acid or
Ac-(0-phosphono1-L-Tvr-L-Ala-N(3-carboxvpropvl1 (3-phenvlpropyll
Figure imgf000056_0001
Step 1: Aminobutanoate-gamma-tert-butyl ester
Aminobutanoate- gamma-tert-butyl ester can be prepared in accordance with methods well known to those skilled in the art. (See, for example, Sluka, et al. ,
J. A er. Chem. Soc. , 1990;112:6369-6374.)
Step 2: 4-(3-Phenyl-propylaminol-butyric acid tert-butyl ester
To a solution of aminobutanoate- gamma-tert-butyl ester in N,N-dimethylformamide (500 mL) at room temperature was added triethylamine (28 mL, 210.2 mmol), followed by dropwise addition of 1-bromo- 3-phenylpropane (15.2 mL, 100.6 mmol). The reaction was stirred overnight at room temperature then concentrated under reduced pressure to 20 mL. The residue was then diluted with ethyl acetate and washed with saturated sodium chloride (3X250 mL) , dried over magnesium sulfate, filtered, and concentrated under reduced pressure to yield a white powder. The powder was dried under reduced pressure to yield (27.0 g,
99%). l-H NMR (400 MHz, DMSO): δ 9.00 (bs, IH), 7.30 (m, 2H), 7.21 (m, 3H) , 2.86 (m, 4H) , 2.65 (t, 2H) , 2.32 (t, 2H), 1.94 (dt, 2H), 1.85 (dt, 2H) , 1.40 (s, 9H) ; Mass Spectrum (Chemical Ionization, 1% NH3 in CH4) m/z 278 (M+H) . Step 3 : Ac- (Q-phosphono 1 -L-Tyr-L-Ala-N ( 3 - propylcarboxy) (3-phenylpropyl)
The title compound was synthesized in a manner similar to that described for Example 1. The product was obtained as a colorless powder (127 mg) . HPLC 100%, rt = 15.1 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 576.7 (M-H).
EXAMPLE 27 rs-(R*,R*1)-5- rr2-f2-Acetylamino-3-(4-phosphonooxy- phenvl1-propionylamino) -propionyl)-(3-phenvl-propyl1 amino) -pentanoic acid or
Ac-fO-phosphono1-L-Tyr-L-Ala-N(4-carboxvbutvl1 (3-phenylpropyll
Figure imgf000057_0001
The title compound was synthesized in a manner similar to that described for Example 1. The product was obtained as a colorless powder (76 mg) . HPLC 100%, rt = 16.1 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 590.3 (M-H).
EXAMPLE 28 rs-(R*,R*1) -Phosphoric acid mono- - (2-acetylamino-2- f1- rbutvl- (3-Phenvl- ropvl ) -carbamoyl) -ethyIcarbamovl) ethyl)-phenyl] ester
Figure imgf000058_0001
The title compound was synthesized in a manner similar to that described for Example 1. The product was obtained as a colorless powder (59 mg) . HPLC 100%, rt = 18.5 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 547.4 (M-H).
EXAMPLE 29 rS- (R* ,R*1 ) -4- r2-Acetylamino-3- (4-phosphonooxy-phenyl ) - propiony amino1-4- \3-(4-carboxymethoxy-phenyl)- propylcarbamovl) -butvric acid or
Ac- (O-phosphono1 -L-Tvr-L-Glu-NHf3- T4- (O-acetic acidl - phenyl1propyl)
Figure imgf000059_0001
The title compound was synthesized in a manner similar to that described for Example 1. The product was obtained as a white solid (60 mg) . HPLC 93%, rt = 12.7 minutes, C18, eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 623.4 (M-H).
EXAMPLE 30 4- r (S1 -2- cetylamino-3- (4-phosphonooxy-phenyl ) - (S1 - fpropionylamino) -5- ( (S ) -2-benzyl-pyrrolidin-1-yl1 -5■ oxo-pentanoic acid or
Ac- (O-phosphonol -L-Tyr-L-Glu-L- Pro t2-decarboxy-2- benzyll
Figure imgf000060_0001
The title compound was synthesized in a manner similar to that described for Example 1. The product was obtained as a colorless solid (167 mg) . HPLC 100%, rt = 15.4 minutes, C18, eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 575.5 (M-H).
EXAMPLE 31 L-Tyrosinamide, N-acetyl-O-phosphono-L-tyrosvl-N- (3 phenylpropyl)-Q-phcsphono- or
Ac- (O-phosphonol -L-Tvr- (0-phosphonol -L-Tyr-NH(3- phenylpropy
Figure imgf000060_0002
The title compound was synthesized in a manner similar to that described for Example 1. The product was obtained as a colorless solid (64 mg) . HPLC 96%, rt = 14.5 minutes, C18, eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 663.3 (M-H).
EXAMPLE 32 4- r (RS1-2-(Acetyl-methvl-amino1-3- (4-phosphonooxy- phenyl)-(S)-propionylamino)-4~(3~phenyl- propylcarbamoyll-butyric acid or
Ac- ( N-methyl O-phosphono l -D/L-Tyr-L-Glu-NH ( 3 - phenylpropyl1
Figure imgf000061_0001
The title compound was synthesized in a manner similar to that described for Example 1. The product was obtained as a colorless solid (52 mg) . HPLC 100%, rt = 14.9 and 15.13 minutes, C18, eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 563.6 (M-H). EXAMPLE 33 rs-(R*,R*1)-4-r2-Methanesulfonylamino-3-(4- phosphonooxv-Phenyl ) -propionvla inol -4- (3-phenyl- propylcarbamovl1 -butyric acid or Methylsulfonyl- (O-Phosphonol -L-Tyr-L-Glu-NH(3- phenvlpropvll
Figure imgf000062_0001
The title compound was synthesized in a manner similar to that described for Example 1. The product was obtained as a colorless powder (71 mg) . HPLC 83%, rt = 15.6 minutes, C18 column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 584.3 (M-H).
EXAMPLE 34 rs-(R*,R*11-r4-(2-Acetylamino-2-fl-r(3-cyclohexyl' propyl1 -methyl-carbamoyl) -propylcarbamoy1) -ethyl1 ■ phenyl) -phosphonic acid or Ac-(4-phosphonvl1-L-Phe-L-Abu-N(methyl1 (3-cyclohexvlpropvl)
Figure imgf000063_0001
Step 1: Ac- f4-(diethoxyphosphonyl1) -L-Phe
Ac- [4-(diethoxyphosphonyl)] -L-Phe was prepared in a manner similar to Ac- [4- (diethoxyphosphonyl)- difluoromethyl] -L-Phe, except starting from Boc- [4- (diethoxyphosphonyl)-difluoromethyl]-L-Phe Benzyl ester, which can prepared in accordance with methods well known to those skilled in the art. (See, for example, Thurieau, et al., J. Med.Chem. , 1994;37:625- 629.) (See Example 5, Step 1.) Deprotection, acetylation, and hydrogenation yielded a solid foam.
^-H NMR (DMSO, 400 MHz): δ 1.22 (t, 6H) , 1.77 (s, 3H) , 2.91 (dd, IH), 3.10 (dd, IH) , 3.33 (m, 4H) , 4.44 (m, IH), 7.38 (dd, 2H) , 7.61 (dd, 2H), 8.22 (d, IH) ; Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 342 (M-H).
Step 2: Ac- (4-phosphonyl1-L-Phe-L-Abu-N(methvl )
(3-cyclohexylpropyl1
The title compound was synthesized in a manner similar to Example 10 except Ac- [4-(phosphonyl)- difluoromethyl]-L-Phe was coupled in rather than Ac- [4- (diethoxyphosphonyl)difluoromethyl] -L-Phe. The purified peptide (30 mg) was deprotected with trimethysilyl bromide (1 mL) in dichloromethane (2 mL) for 4 hours at room temperature. Water (1 mL) and trifluoroacetic acid (1 mL) were added to quench excess trimethysilyl bromide, and the resulting solution was concentrated at reduced pressure to remove volatiles. An additional 1 mL of TFA was added, and the mixture was concentrated to -200 μL and then precipitated with diethyl ether. The resulting solid was filtered, washed with diethyl ether, and dried under vacuum overnight to yield 27 mg of an off-white solid. HPLC 98%, rt = 18.6 minutes, C18, eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 508 (M-H).
EXAMPLE 35 (S1 -4- r3-Carboxvmethvl-3-(4-phosphonooxy-benzyl1- ureidol -4- f (3-cyclohexyl-propyll-methyl-carbamoyl)- butyric acid
Figure imgf000064_0001
To Glu(OtBu)-N(Me)-3-cyclohexylpropyl (360 mg, 1.0 mmol) synthesized as an intermediate in Example 1, ice, dichloromethane (20 mL), and saturated sodium bicarbonate (20 mL) in a separatory funnel was added phosgene in toluene (7.5 equiv. ) . The mixture was shaken for 5 minutes, then the organic layer was separated, dried over magnesium sulfate, and concentrated to give the isocyanate as a colorless oil The isocyanate was treated in dichloromethane (20 mL) with 4-hydroxybenzyl glycine-t-butyl ester (250 mg, 1.0 mmol) and triethylamine (0.15 mL, 1.0 mmol) for 1 hour. Ethyl acetate was added, and the reaction mixture was washed with 10% sulfuric acid, then water, and then brine. After drying over magnesium sulfate, the solvent was removed under reduced pressure to give then urea as a colorless oil (500 mg, 83%). The crude urea was phosphorylated, oxidized, deprotected, and purified in a manner similar to Example 1, giving the title compound (49 mg) as a colorless solid. HPLC 100%, rt = 17.4 minutes, C18, eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA, and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 552.4 (M-H).
EXAMPLE 36 (SI-4- r (3-Cyclohexyl-propyl1 -methyl-carbamoyl) -4- T3-(6- phosphonoxy-naphthalen-1-yll-ureido) -butvric acid
Figure imgf000065_0001
Step 1: (S1-2-r3-(6-Hydroxy-naphthalen-l-yl1- ureido) -pentanedioic acid 5-tert-butvl ester 1-methyl ester
To H-Glu(OtBu)-OCH3 (2.0 mmol, 0.50 g) in dichloromethane (25 mL) at 0°C was added pyridine (8 mmol, 0.66 mL) then followed by dropwise addition of phosgene in toluene (3.0 mmol, 1.60 mL, 1.92 M) . After stirring at 0°C for 2 hours, 1-amino-6-naphthol (2.0 mmol, 0.32 g) was added in dichloromethane. After stirring at room temperature for 12 hours, the reaction mixture was evaporated to dryness and purified on silica gel (1:19, methanol/dichloromethane) to provide product as a light brown solid (0.71 g, 88%).
Step 2; (S) -2- r3-(6-Hvdroxy-naphthalen-l-yll- ureido)-pentanedioic acid 5-tert-butyl ester
To the methyl ester from Step 1 above (5 mmol, 2.0 g) was added potassium hydroxide (5 mmol, 0.28 g) in water (7 L) , and heated at 50°C. After stirring for 36 hours, the reaction mixture was evaporated to dryness, the residue was dissolved in water (15 mL), acidified to pH »5 using 0.50N hydrochloric acid, extracted with the ethyl acetate (4X50 mL), and dried to provide product as a light brown oil (1.20 g, 62%).
Step 3: (S )-4- f (3-Cyclohexyl-propyl1- ethyl- carbamoyl) -4- T3-(6-hydroxv-naphthalen-l-yll-ureido) - butyric acid tert-butvl ester
This step was synthesized 'in a manner similar to that described for Example 1 (Step 1) .
Step 4: (SI-4- (3-Cyclohexyl-propyl -methyl- carbamoyl) -4- Ϊ 3 -(6-phosphonoxγ-naphthalen-l-yl ureidol-butyric acid
The title compound was synthesized in a manner similar to that described for Example 1. Product was obtained as a white solid (27 mg) . HPLC 100%, rt = 18.9 minutes, C18, eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 549.4 (M-H). EXAMPLE 37 rs- (R* ,R*1 ) -4- r2-Acetylamino-3- (4-phosphonooxv-phenvl) propionylamino) -4- (5-methyl-hexylcarba oyll -butyric acid or Ac-(0-phosphono1-L-Tyr-L-Glu-N(5-methylhexyl1
Figure imgf000067_0001
The title compound was synthesized in a manner similar to that described for Example 1. The product was obtained as a colorless solid (154 mg) . HPLC 95%, rt = 15.2 minutes, C18, eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 529.6 (M-H).
EXAMPLE 38 rs- (R*,R ) -4- r2-Acetylamino-3- (4-phosphonooxy-phenyl1 - propionylamino) -4- (4-methyl-hexylcarbamoyll -butyric acid or
Ac-(0-phosphono1-L-Tvr-L-Glu-NH((R,S1-4-methylhexyl1
Figure imgf000068_0001
The title compound was synthesized in a manner similar to that described for Example 1. The product was obtained as a colorless solid (91 mg) . HPLC 94%, rt = 15.1 minutes, C18, eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 528.5 (M-H).
EXAMPLE 39 Phosphoric acid mono- -T4-(4-benzyloxy-3-carbamoyl- benzylcarbamovl )-phenvl) ester
Figure imgf000068_0002
The title compound was synthesized in a manner similar to that described for Example 44. The product was obtained as a colorless powder (21 mg) . HPLC 100%, rt = 14.8 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 455.2 (M-H).
EXAMPLE 40 Phosphoric acid mono- f4- r3-carbamoyl-4- (3-methyl- benzyloxv1 -benzylcarbamoyl1 -phenyl) ester
Figure imgf000069_0001
The title compound was synthesized in a manner similar to that described for Example 44. The product was obtained as a colorless powder (73 mg) . HPLC 100%, rt = 16.9 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 469.3 (M-H).
EXAMPLE 41 Phosphoric acid mono-f4- r3-carbamoyl-4- (3 , 5-dimethyl- benzyloxy ) -benzylcarbamoyl) -phenyl) ester
Figure imgf000070_0001
The title compound was synthesized in a manner similar to that described for Example 44. The product was obtained as a colorless powder (85 mg) . HPLC 100%, rt = 17.2 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 483.4 (M-H).
EXAMPLE 42 Phosphoric acid mono-f4- r3-carbamoyl-4-(4-methyl- pentyloxy1-benzylcarbamovl) -phenyl) ester
Figure imgf000070_0002
The title compound was synthesized in a manner similar to that described for Example 44. The product was obtained as a colorless powder (8 mg) . HPLC 100%, rt = 18.7 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 449.4 (M-H).
EXAMPLE 43 Phosphoric acid mono-f4- r3-carbamoyl-4- (4-methyl- hexyloxvl -benzylcarbamoyl) -phenyl) ester
Figure imgf000071_0001
The title compound was synthesized in a manner similar to that described for Example 44. The product was obtained as a colorless powder (100 mg) . HPLC 99%, rt = 18.3 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 463.4 (M-H).
EXAMPLE 44
Phosphoric acid mono- - ( -carbamoyl-4-cvclohexvl■ methoxv-benzylcarbamovl1 -phenyl ) ester
Figure imgf000072_0001
Step 1; 5-Aminomethvlsalicvlic acid 5-Aminomethylsalicylic acid can prepared in accordance with methods well known to those skilled in the art. (See, for example, Sekiya, et al., Chem. Pharm. Bull. , 1963;11:551-553.)
Step 2; 5-(tert-Butoxycarbonylamino-methyll-2- hydroxy-benzoic acid
To a solution of 5-aminomethylsalicyclic acid (8.3 g, 49.7 mmol) in 100 mL water and 100 mL of dioxane at 0°C was added IN sodium hydroxide (54.7 mL, 54.7 mmol) followed by di-tert-butyl dicarbonate (11.9 g, 54.7 mmol). The reaction was allowed to warm slowly to room temperature and stirred overnight. The reaction was then concentrated under reduced pressure to 15 mL. A layer of ethyl acetate was then added, and the reaction was acidified to pH 2 with saturated potassium hydrogen sulfate. The aqueous layer was then extracted with ethyl acetate (3X150 mL) . The combined ethyl acetate layers were combined and washed with saturated sodium chloride (1X150 mL) , dried over magnesium sulfate, filtered, and concentrated under reduced pressure to yield a white powder. The powder was collected and washed with ether, followed by drying under reduced pressure to yield (11.5 g, 87%). 1H NMR (400 MHz, DMSO): δ 7.67 (s, IH) , 7.38 (d, 2H), 6.91 (d, IH), 4.04 (d, 2H), 1.41 (s, 9H) . Mass Spectrum (Chemical Ionization, 1% NH3 in CH4) m/z 268 (M+H) . Step 3: (3-Carbamoyl-4-hydroxy-benzyl -carbamic acid tert-butvl ester
To a solution of 5-(tert-Butoxycarbonylamino- methyl) -2-hydroxy-benzoic acid (20.0 g, 74.9 mmol) in 700 mL tetrahydrofuran at 0°C was added 4-methyl morpholine (12.3 mL, 112.3 mmol), 1-hydroxybenzo- triazole (15.2 g, 112.3 mmol), followed by l-(3- dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (21.5 g, 112.3 mmol). Stir for 1 hour at 0°C, then added concentrated ammonium hydroxide (15.2 mL,
112.3 mmol). The reaction was allowed to warm slowly to room temperature and stirred overnight. The reaction was then diluted with ethyl acetate and washed with 5% citric acid (3X250 L) , saturated sodium bicarbonate (3X250 mL) , and saturated sodium chloride
(1X250 mL) , dried over magnesium sulfate, filtered, and concentrated under reduced pressure to yield a pale yellow powder. The powder was collected and washed with ether, followed by drying under reduced pressure to yield (14.0 g, 71%). 1H NMR (400 MHz, DMSO): δ 12.77 (bs, IH), 8.33 (bs, IH) , 7.72 (d, IH) , 7.28 (d, 2H), 6.84 (d, IH), 4.05 (d, 2H) , 1.38 (s, 9H) ; Mass Spectrum (Chemical Ionization, 1% NH3 in CH4) m/z 267 (M+H) . Step 4; (3-Carbamoyl-4-cyclohexylmethoxy-benzyl ) - carbamic acid tert-butyl ester
To a solution of (3-Carbamoyl-4-hydroxy-benzyl)- carbamic acid tert-butyl ester (2.0 g, 7.5 mmol) in 15 mL methanol at room temperature was added cesium carbonate (2.7 g, 8.3 mmol). The reaction was stirred overnight. The reaction was then concentrated under reduced pressure. N,N-dimethylformamide (50 mL) was then added, and the reaction was then reconcentrated under reduced pressure to remove any remaining methanol. The residue was then suspended in N,N- dimethylformamide (15 mL) and cyclohexylmethyl bromide (1.1 mL, 8.3 mmol) was then added, and the reaction was heated at 65°C for 4 hours. The reaction was then concentrated under reduced pressure. The residue was diluted with ethyl acetate and 5% citric acid. The organic layer was washed with 5% citric acid (2X50 mL) , IN sodium hydroxide (2X50 mL) , 5% citric acid (2X50 mL) , and saturated sodium chloride (1X50 mL), dried over magnesium sulfate, filtered, and concentrated under reduced pressure to yield a pale yellow powder. The powder was collected and washed with ether, followed by drying under reduced pressure to yield (1.5 g, 57%). 1H NMR (400 MHz, DMSO): δ 7.68 (bs, IH), 7.52 (d, 2H) , 7.36 (t, IH) , 7.27 (d, 2H) , 7.05 (d, IH), 4.04 (d, 2H) , 3.88 (d, 2H) , 1.76-1.61 (m, 6H), 1.36 (s, 9H), 1.28-1.02 (m, 5H) ; Mass
Spectrum (Chemical Ionization, 1% NH3 in CH4) m/z 363 (M+H) .
Step 5; Benzamide, 2-(cyclohexylmethoxy )-5- T f f (4- hvdroxvphenvl1-carbonvl1 mino) -methyl) A solution of 4N hydrochloric acid in dioxane was added to (3-Carbamoyl-4-cyclohexylmethoxy-benzyl) - carbamic acid tert-butyl ester (1.0 g, 2.7 mmol). The reaction is stirred for 1 hour and then concentrated under reduced pressure to yield a solid. This solid of the amine hydrochloride was then used for the coupling. The above amine hydrochloride was dissolved in N,N- dimethylformamide (30 L), and the solution was cooled to 0°C. 4-methyl morpholine (0.75 mL, 6.9 mmol) was then added, followed by 1-hydroxybenzotriazole (559 mg, 4.1 mmol), 4-hydroxybenzoic acid (457 mg, 3.3 mmol), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (793 mg, 4.1 mmol). The reaction was allowed to warm slowly to room temperature and stirred overnight. The reaction was concentrated under reduced pressure in 10 mL. The residue was then diluted with ethyl acetate and washed with 5% citric acid (3X250 mL) , saturated sodium bicarbonate (3X250 mL) , and saturated sodium chloride (1X250 mL) , dried over magnesium sulfate, filtered, and concentrated under reduced pressure to yield a pale yellow powder. The powder was collected and washed with ether, followed by drying under reduced pressure to yield (871 mg, 83%). 1H NMR (400 Hz, DMSO): δ 9.97 (s, IH) , 8.78 (t, IH) , 7.75 (m, 3H), 7.58 (d, 2H), 7.38 (d, IH) , 7.05 (d, IH) , 6.78 (d, 2H), 4.35 (d, 2H) , 3.88 (d, 2H) , 1.61-1.77 (m, 6H), 1.15-1.05 (m, 5H) ; Mass Spectrum (Chemical Ionization, 1% NH3 in CH4) m/z 383 (M+H) .
Step 6: Phosphoric acid mono- -(3-carbamoyl-4- cyclohexylmethoxy-benzvlcarbamovl1 -phenyl) ester
The title compound was synthesized in a manner similar to that described for Example 1. The product was obtained as a colorless powder (50 mg) . HPLC 92%, rt = 17.2 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 461.5 (M-H).
EXAMPLE 45 r4- (3-Carbamoyl-4-cyclohexvlmethoxy-benzylcarbamoy11- phenvl) -difluoro-methvl-phosphonic acid
Figure imgf000075_0001
Step 1; fPifluoro-(4-iodo-phenyll-methyl) - phosphonic acid diethyl ester
[Difluoro-(4-iodo-phenyl)-methyl] -phosphonic acid diethyl ester can be prepared in accordance with methods well known to those skilled in the art. (See, for example, Burke, et al. , J. Org. Chem. , 1993;58: 1336-1340. )
Step 2; fDifluoro- (4-formvl-phenyl )-methyl) - phosphonic acid diethyl ester To a solution of [Difluoro-(4-iodo-phenyl)- methyl] -phosphonic acid diethyl ester (1.6 g, 4.1 mmol) in anhydrous ether (40 mL) at -78°C was added dropwise n-butyl lithium (2.5 M in hexanes, 2.5 mL, 6.1 mmol). The brown solution was stirred at -78°C for 2 minutes. Ethyl formate (0.66 mL, 8.2 mmol) was then added, and the reaction was stirred for 10 minutes at -78°C. The mixture was then quenched with saturated ammonium chloride (10 mL) and warmed to room temperature. The reaction was then diluted with ether. The organic layers were washed with saturated sodium chloride
(20 mL), dried over magnesium sulfate, filtered, and concentrated under reduced pressure to yield a yellow oil. Chromatography of the residue [3:7, ethyl acetate:hexanes] gradient to (1:1)] afforded the aldehyde (268 mg, 22%) as a clear oil.
^■H NMR (400 MHz, DMSO): δ 10.28 (s, IH) , 8.25 (d, 2H) , 7.97 (d, 2H), 4.42 (m, 4H) , 1.40 (t, 6H) .
Step 3; 4- \(Diethoxy-phosphoryl)-difluoro¬ methyl) -benzoic acid To a solution of [Difluoro-(4-formyl-phenyl)- methyl] -phosphonic acid diethyl ester (260 mg, 0.9 mmol) in acetone (25 mL) at room temperature was added Jones reagent (5 mL) . The reaction was quenched by addition of ethanol (5 mL) . The reaction was then concentrated under reduced pressure to 5 mL. The residue was diluted with ethyl acetate and washed with water (2X20 mL), saturated sodium chloride (20 mL) , dried over magnesium sulfate, filtered, and concentrated under reduced pressure to yield a white crystalline solid (268 mg, 98%). 1H NMR (400 MHz, DMSO): δ 8.09 (d, 2H) , 7.70 (d, 2H) , 4.15 (m, 4H) ,
1.22 (t, 6H); Mass Spectrum (Chemical Ionization, 1% NH in CH4) m/z 309 (M+H) .
Step 4: r4- (3-Carbamoyl-4-cyclohexylmethoxy- benzylcarbamoyll -phenyl) -difluoro-methyl-phosphonic acid
The title compound was synthesized in a manner similar to that described for Example 44. The product was obtained as a colorless powder (39 mg) . HPLC 100%, rt = 17.4 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 495.4 (M-H).
EXAMPLE 46
Phosphoric acid mono- r4- (3-carbamoyl-4-cyclohexyl- methoxy-benzylcarbamoyl1 -2, 6-dimethyl-phenyl) ester
Figure imgf000077_0001
The title compound was synthesized in a manner similar to that described for Example 44. The product was obtained as a colorless powder (79 mg) . HPLC 100%, rt = 18.3 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 489.4 (M-H).
EXAMPLE 47 Phosphoric acid mono- f4- (3-carbamoyl-4-cyclohexyl- methoxy-benzylcarbamoyll-2-chloro-phenyl) ester
Figure imgf000078_0001
The title compound was synthesized in a manner similar to that described for Example 44. The product was obtained as a colorless powder (119 mg) . HPLC 100%, rt = 17.7 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 495.3 (M-H).
EXAMPLE 48 Phosphoric acid mono- f4-(4-cvclohexvlmethoxv-3- methylcarbamoyl-benzvlcarbamoyll-phenyl1 ester
Figure imgf000079_0001
The title compound was synthesized in a manner similar to that described for Example 44. The product was obtained as a colorless powder (79 mg) . HPLC 100%, rt = 18.3 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 475.5 (M-H).
EXAMPLE 49 (SI-Phosphoric acid mono-f4- r2-acetylamino-2-(3- carbamoyl-4-cyclohexyImethoxy-benzylcarbamoyl1-ethyl) - phenyl) ester
Figure imgf000079_0002
The title compound was synthesized in a manner similar to that described for Example 44. The product was obtained as a colorless powder (60 mg) . HPLC 99%, rt = 16.9 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 546.4 (M-H).
EXAMPLE 50 (SI-Phosphoric acid mono-(4-f2-acetylamino-2- T3- carbamovl-4-(3,5-dimethvl-benzvloxy1-benzylcarbamoyl)- ethyl)-phenyl1 ester
Figure imgf000080_0001
The title compound was synthesized in a manner similar to that described for Example 44. The product was obtained as a colorless powder (59 mg) . HPLC 98%, rt = 17.0 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 568.4 (M-H).
EXAMPLE 51 Phosphoric acid mono-(4-f (SI -2-acetylamino-2- r (RSI-1- (3-carbamoyl-4-cvclohexvl ethoxv-phenvl1- ethylcarbamoyl1 -ethyl)-phenyl) ester
Figure imgf000081_0001
The title compound was synthesized in a manner similar to that described for Example 44. The product was obtained as a colorless powder (61 mg) . HPLC 99%, rt = 19.7 and 20.10 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes.
Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 560.4 (M-H).
Alternatively, the title compound can be synthesized as follows: Step 1: 5-Acetyl-2-hydroxy-benzamide
The title compound was synthesized in a manner similar to that described for Example 44, Step 3. The product was obtained as a solid. 1H NMR (400 MHz, DMSO): 6 13.84 (s, IH) , 8.73 (bs, IH) , 8.53 (s, IH) , 8.12 (bs, IH), 8.00 (d, IH) , 6.99 (d, IH), 2.59
(s, 3H) . Mass Spectrum (Chemical Ionization, 1% NH3 in CH4) m/z 179 (M+H) .
Step 2; 5-Acetyl-2-cvclohexylmethoxy-benzamide
The title compound was synthesized in a manner similar to that described for Example 44, Step 4. The product was obtained as a solid. 1H NMR (400 MHz, DMSO): δ 8.31 (s, IH) , 8.05 (d, IH) , 7.22 (d, IH) , 4.04 (d, 2H), 2.53 (s, 3H) , 1.83 (m, 6H), 1.14 (m, 5H) .Mass Spectrum (Chemical Ionization, 1% NH3 in CH4) m/z 275 (M+H) . Step 3: (E/Z1-2-Cyclohexylmethoxy-5-(l- hydroxvimino-ethyl1-benzamide
To a solution of 5-acetyl-2-cyclohexylmethoxy- benzamide (1.0 g, 3.63 mmol) in pyridine (25 mL) at room temperature was added hydroxylamine hydrochloride (0.38 g, 5.45 mmol). The reaction was stirred 48 hours. The reaction was then concentrated under reduced pressure, diluted with ethyl acetate and washed with 5% citric acid (3X100 mL), saturated sodium chloride (1X100 mL), dried over magnesium sulfate, filtered, and concentrated under reduced pressure to yield a foam. ^-H NMR (400 MHz, DMSO): δ 11.07 (s, IH), 8.08 (d, IH), 7.72 (d, IH) , 7.61 (bs, IH) , 7.53 (bs, IH), 7.14 (d, IH) , 4.03 (d, 2H) , 2.13 (s, 3H), 1.74 (m, 6H) , 1.11 (m, 5H) . Mass Spectrum (Chemical Ionization, 1% NH3 in CH4) m/z 291 (M+H).
Step 4; (+/-1-5-(l-Amino-ethyl1-2- cyclohexvlmethoxy-benzamide
To a solution of (E/Z)-2-Cyclohexylmethoxy-5-(1- hydroxyimino-ethyl)-benzamide (0.50 g, 1.72 mmol) in methanol (20 mL) and triethylamine (5 mL) was added wet Raney Nickel (0.30 g) under hydrogen at 45 psi for 16.5 hours. The reaction was then concentrated under reduced pressure to yield a white solid. 1H NMR (400 MHz, DMSO): δ 7.86 (s, IH) , 7.59 (d, 2H) , 7.47 (d, IH), 7.10 (d, IH), 4.03 (q, IH) , 3.97 (d, 2H) , 1.83 (m, 6H), 1.23 (m, 8H) . Mass Spectrum (Chemical Ionization, 1% NH3 in CH4) m/z 277 (M+H).
Phosphoric acid mono-(4-f (S)-2-acetvlamino-2- T (RS1-1-(3-carbamoyl-4-cyclohexylmethoxv-phenvl )- ethylcarbamoyll -ethyl)-phenvl1 ester
The title compound was synthesized in a manner similar to that described for Example 44. The product was obtained as a colorless powder (61 mg) . HPLC 99%, rt = 19.7 and 20.1 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 560.4 (M-H).
EXAMPLE 52 (SI-Phosphoric acid mono-14- r2-acetylureido-2-(3- carbamoyl-4-cyclohexylmethoxy-benzylcarbamoyll-ethyl) phenvl) ester
Figure imgf000083_0001
Step 1: (S1-3-(4-Benzyloxy-phenyl1-2-(3-methyl- ureido)-propionic acid benzyl ester This compound was synthesized in a manner similar to that described in Example 7 (Step 1).
Step 2; (S1-3- (4-Hydroxy-phenyl )-2- (3-methvl- ureidol-propionic acid
This compound was synthesized in a manner similar to that described in Example 7 (Step 2) .
Step 3: (S )-2-Cyclohexylmethoxy-5-f \3-(4-hydroxy¬ phenyl1-2-(3-methvl-ureidol-propionylamino) -methyl)- benaamide
This compound was synthesized in a manner similar to that described in Example 44 (Step 2).
Step 4: (SI-Phosphoric acid mono-f4-r2- acetylureido-2-(3-carbamovl-4- cyclohexylmethoxvbenzyIcarbamovl1-ethvl) -phenvl)ester The title compound was synthesized in a manner similar to that described for Example 1. Product was obtained as a white solid (100 mg) . HPLC 98%, rt = 17.4 minutes, C18, eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 561.4 (M-H).
EXAMPLE 53 (S )- 4- r2-Acetylamino-2- (3-carbamoyl-4-cyclohexyl• methoxv-benzyIcarbamovl1-ethyl) -phenyl)-difluoro¬ methyl-phosphonic acid
Figure imgf000084_0001
The title compound was synthesized in a manner similar to that described for Example 49 and cleaved according to Example 5 (Step 2) . The product was obtained as a colorless powder (3.8 mg) . HPLC 97%, rt = 17.6 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 580.5 (M-H). EXAMPLE 54 rS- (R* ,R*11 -4- 2-Acetylamino-3- (4-phosphono-phenyl1 - propionylamino) -4- f (3-cyclohexyl-propyl1 -methyl- carbamoyl) -butyric acid or
Ac-(4-phosphonyl1-L-Phe-L-Glu-N(methyl1 (3- cyclohexvlpropvl )
Figure imgf000085_0001
The title compound was synthesized in a manner similar to that described for Example 34. The product was obtained as a colorless powder (125 mg) . HPLC 100%, rt = 12.4 minutes, C18, column eluting with a gradient of 0% to 100% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 20 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 552 (M-H).
EXAMPLE 55 rs-(R*,R»11-2,2-Dimethyl-propionic acid M4-(2- acetyla ino-2-f1- r( -cyclohexyl-propyl)-methyl- carbamoyl) -propvlcarbamoγl) -ethyl1 -phenvl) -difluoro¬ methyl)-hydroxv-phosphinoyloxymethyl ester
Figure imgf000086_0001
Ac-(4-(difluorophosphonomethyl11-L-Phe-L-Abn-
N(methyl1 (3-cyclohexylpropyl ) (from Example 10) (0.15 mmol, 84 mg) was dissolved in 2 mL dimethylformamide and treated with diisopropylethylamine (0.5 mmol, 81 μL) and chloromethyl pivaloate (1.5 mmol, 214 μL) . The reaction mixture was stirred at 70 °C for 24 hours. The reaction mixture was diluted with 3 mL of dimethylformamide and purified by preparative HPLC, as previously described, to provide the product as a colorless solid after lyophilization (58 mg) . HPLC 100%, rt = 14.8 minutes, C18, column eluting with a gradient of 0% to 100% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 20 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 672 (M-H).
EXAMPLE 56 rs-(R*,R*11-4- r2-Acetylamino-3-(4-phosphonooxy-phenyl1- propionylamino)-4-(2-adamantan-l-yl-ethvlcarbamoyl1- butyric acid
Figure imgf000087_0001
The title compound was synthesized in a manner similar to that described for Example 1. The product was obtained as a white solid (50 mg) . HPLC 100%, rt = 18.0 minutes, C18, eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water +0.1% ammonium hydroxide) m/z 593 (M-H) .
EXAMPLE 57
(S)-Phosphoric acid mono-(4-f2-acetylamino-2- ri-ethyl-
1-(3-phenyl-propylcarbamoyl1-propylcarbamoyl) -ethyl)- phenyl1 ester
Figure imgf000087_0002
The title compound was synthesized in a manner similar to that described for Example 1. The product was obtained as a white solid (150 mg) . HPLC 100%, rt = 18.0 minutes, C18, eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water +0.1% ammonium hydroxide) m/z 532 (M-H) .
EXAMPLE 58 rs-(R*,R ) -Phosphoric acid mono- (4-f2-acetylamino-2- r2-hvdroxv-l-(3-phenyl-propylcarbamoyl1- propvlcarbamoyl1 -ethyl)-phenvl ) ester
Figure imgf000088_0001
The title compound was synthesized in a manner similar to that described for Example 1. The product was obtained as a white solid (46 mg) . HPLC 100%, rt = 13.8 minutes, C18 , eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water +0.1% ammonium hydroxide) m/z 520 (M-H) .
EXAMPLE 59 TS- (R* ,R*1) -4- r (3-Cvclohexvl-propyl1 -methyl-carbamoyl) - 4- r2- (3, 3-dimethyl-ureido1 -3- (4-phosphonooxy-phenvl1 - propionylamino) -butvric acid or
N- r (Dimethylaminolcarbonyl) - (0-phosphonol -L-Tyr-L-Glu- N(methyl1 (3-cyclohexvlpropvl1
Figure imgf000089_0001
The title compound was synthesized in a manner similar to that described for Example 7. The product was obtained as a colorless solid (110 mg) . HPLC 100%, rt = 17.9 minutes, C18, eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water +0.1% ammonium hydroxide) m/z 597 (M-H).
EXAMPLE 60 5- r (3-cyclohexvlpropvl1methylamino)-4-r f2- r r (dimethylamino1 -sulfonvl)amino1 -1-oxo-3- T4- (phosphonooxylPhenyl)propyl) mino) -5-oxo-pentanoic acid
Figure imgf000089_0002
The title compound was synthesized in a manner similar to that described for Example 33. The product was obtained as a white solid (115 mg) . HPLC 100%, rt = 20.7 minutes, C18, eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water +0.1% ammonium hydroxide) m/z 633 (M-H).
EXAMPLE 61 (S -4- f (3-Cvclohexvl-propvl \-methyl-carbamoyl) -4- r3- (4- phosphonooxv-benzvH-ureido) -butyric acid
Figure imgf000090_0001
The title compound was synthesized in a manner similar to that described in Example 35. The product was obtained as a colorless solid (47 mg) . HPLC 98%, rt = 17.8 minutes, C18, eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA, and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 512 (M-H) .
EXAMPLE 62 (S^-4- r (3-Cyclohexyl-propyH-methyl-carbamoyl) -4-(4- phosphonooxy-benzoylamino^-butyric acid
Figure imgf000091_0001
The title compound was synthesized in a manner similar to Example 1 and 39. The product was obtained as a colorless solid (70 mg) . HPLC 95%, rt = 17.6 minutes, C18, eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water +0.1% ammonium hydroxide) m/z 483 (M-H).
EXAMPLE 63a rs-(R*,R*n- or rs-(R*,S* -r(4-f2-Acetylamino-2-ri-(3- carbamoyl-4-cyclohexylmethoxy-phenyll-ethylcarbamoyl) - ethvl)-Phenyl^-difluoro-methyl) -phosphonic acid
Figure imgf000091_0002
Isomer- 1
The title compound was synthesized in a manner similar to that described for Example 51 and 53, except in this case the mixture of diastereomers was separated by preparative HPLC as previously described. The product was obtained as a colorless powder (31 mg) . HPLC 100%, rt = 12.6 minutes, C18, column eluting with a gradient of 0% to 100% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 20 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 594 (M-H).
EXAMPLE 63b rs-(R*,R*)l- or rs-(R*,S*)1-r(4-f2-Acetylamino-2-n-(3- carbamoyl-4-cyclohexylmethoxy-phenyl)-ethylcarbamoyll- ethvl)-phenvl )-difluoro-methyl) -phosphonic acid
Figure imgf000092_0001
Isomer 2
The title compound is the second isomer separated in Example 51 and 53. The product was obtained as a colorless powder (34 mg) . HPLC >95%, rt = 12.8 minutes, C18, column eluting with a gradient of 0% to 100% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 20 minutes.
Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 594 (M-H).
EXAMPLE 64a Phosphoric acid mono- T4-(2-acetylamino-2-f1- r3- carbamoy1-4-(2-cyclohexyl-1-methyl-ethoxv)- henvl) - ethylcarbamoy1}- thyl)-phenyl1 ester
Figure imgf000093_0001
The title compound was synthesized in a manner similar to that described for Example 51, except in this case the mixture of diastereomers was separated by preparative HPLC as previously described. The product was obtained as a colorless powder (5 mg) . HPLC 100%, rt = 13.1 minutes, C18, column eluting with a gradient of 0% to 100% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 20 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 588 (M-H).
EXAMPLE 64b Phosphoric acid mono- T4-(2-acetylamino-2-f1- f3- carbamoy1-4-(2-cyclohexyl-1-methvl-ethoxy )- henyl1 - ethvlcarbamoyl)-ethyl )-phenvl) ester
Figure imgf000093_0002
The title compound was synthesized in a manner similar to that described for Example 51, except in this case the mixture of diastereomers was separated by preparative HPLC as previously described. The product was obtained as a colorless powder (4 mg) . HPLC 100%, rt = 13.3 minutes, C18, column eluting with a gradient of 0% to 100% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 20 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 588 (M-H).
EXAMPLE 65
( S ) -Phosphoric acid mono- (4- f2-acetylamino-2- (RS) -1- (3-carbamovlmethvl-4-cvclohexylmethoxy-phenyl) - ethyIcarbamovl) -ethvl)-phenyl) ester
Figure imgf000094_0001
Step 1; (2-hydroxyphenyl) -acetic acid methyl
To a solution of 2-hydroxyphenylacetic acid (20.0 g, 129.7 mmol) in 250 L of methanol was bubbled at room temperature anhydrous hydrochloric acid for 5 minutes . The reaction was capped and stirred overnight. The reaction was then concentrated under reduced pressure to yield a pale oil (20.5 g, 95%). 1H NMR (300 MHz, DMSO): δ 9.45 (s, IH) , 7.15 (m, 2H) , 6.75 (m, 2H), 3.57 (d, 2H) , 3.35 (s, 3H) .
Step 2: (2-methoxyphenyl -acetic acid methvl ester
To a solution of (2-hydroxyphenyl) -acetic acid methyl ester (20.0 g, 120.5 mmol) in 200 mL N,N- dimethylformamide was added freshly ground potassium carbonate (24.9 g, 180.7 mmol) followed by iodomethane (8.25 mL, 132.5 mmol). The reaction mixture was heated to 50°C overnight. The reaction was then concentrated under reduced pressure. The residue was diluted with ethyl acetate and water. The organic layer was washed with IN hydrochloric acid (2X100 L) and saturated sodium chloride (1X100 mL), dried over magnesium sulfate, filtered, and concentrated under reduced pressure. Chromatography of the residue (1:4, ethyl acetate/hexanes) gave the product as a clear oil. 1H NMR (400 MHz, DMSO): δ 7.25 (t, IH) , 7.17 (d, IH) , 6.97 (d, IH), 6.89 (t, IH) , 3.74 (s, 3H) , 3.58 (s, 5H) . Mass Spectrum (Chemical Ionization, 1% NH3 in CH4) m/z 180 (M+H) . Step 3; (5-acetvl-2-hydroxyphenyl)-acetic acid methyl ester
To a solution of aluminum chloride (35.0 g, 262.5 mmol) in dichloromethane (75 mL) was added a solution of (2-Methoxyphenyl)-acetic acid methyl ester (13.5 g, 75.0 mmol) and acetyl chloride (5.86 mL,
82.5 mmol)in dichloromethane (75 mL) dropwise. After the addition was complete the reaction was heated to reflux for 5 hours. The reaction mixture was then cooled and carefully poured onto a mixture of ice and water. The aqueous layer was then extracted with ethyl acetate (2X400 mL) . The combined extracts were then washed with saturated sodium chloride (1X100 mL) , dried over magnesium sulfate, filtered, and concentrated under reduced pressure to yield a solid. 1H NMR (400 MHz, DMSO): δ 10.51 (s, IH) , 7.79 (m, 2H) , 6.88 (d, IH), 3.63 (s, 2H), 3.59 (s, 3H), 2.47 (s, 3H) . Mass Spectrum (Chemical Ionization, 1% NH3 in CH4) m/z 209 (M+H) .
Step 4: (5-acetyl-2-cyclohexylmethoxyphenyl)- acetic acid methvl ester This compound was synthesized in a manner similar to that described for Example 44, Step 4. The product was obtained as an oil (7.9 g) . 1H NMR (400 MHz, DMSO): δ 7.89 (d, IH) , 7.84 (s, IH) , 7.05 (d, IH) , 3.86 (d, 2H), 3.66 (s, 2H) , 3.58 (s, 3H) , 2.49 (s, 3H) , 1.81 (m, 6H), 1.12 (m, 5H) . Mass Spectrum (Chemical Ionization, 1% NH3 in CH4) m/z 305 (M+H).
Step 5 ; ( 5-Acetvl - 2-cvclohexylmethoxy phenyl ) - acetic acid To a solution of (5-Acetyl-2-cyclohexylmethoxy phenyl)-acetic acid methyl (7.9 g, 25.9 mmol) in tetrahydrofuran (150 mL) and methanol (150 mL) was added a IN solution of sodium hydroxide (52.0 mL, 52 mmol). The reaction was stirred at room temperature overnight. The reaction was then concentrated under reduced pressure, diluted with ethyl acetate and acidified to pH 2 with IN hydrochloric acid. The aqueous layer was extracted with ethyl acetate (3X200 mL) . The combined extracts where then washed with saturated sodium chloride (1X200 mL), dried over magnesium sulfate, filtered, and concentrated under reduced pressure. 1H NMR (400 MHz, DMSO): δ 12.23 (s, IH), 7.89 (d, IH), 7.81 (s, IH) , 7.04 (d, IH) , 3.86 (d, 2H), 3.56 (s, 2H), 2.49 (s, 3H) , 1.71 (m, 6H) , 1.16 (m, 5H) . Mass Spectrum (Chemical Ionization, 1% NH3 in CH4) m/z 290 (M+H) . step 6: 2-( -Acetyl- -cyclohexyl ethoxy- henyl)- acetamide
This compound was synthesized in a manner similar to that described for Example 44, Step 3. The product was obtained as a solid. 1H NMR (400 MHz, DMSO): δ
7.85 (d, IH), 7.77 (s, IH) , 7.33 (bs, IH) , 7.01 (bs,
IH), 6.89 (s, IH), 3.85 (d, 2H) , 3.42 (s, 2H), 2.50
(s, 3H), 1.72 (m, 6H), 1.16 ( , 5H) . Mass Spectrum (Chemical Ionization, 1% NH3 in CH4) m/z 289 (M+H). Step 7: (E/Z) -2- (2-cyclohexvlmethoxv-5- (1- hydroxvimino-ethyl)-phenyl) -acetamide
This compound was synthesized in a manner similar to that described for Example 51, Step 3. The product was obtained as a solid. 1H NMR (400 MHz, DMSO): δ 10.92 (s, IH), 7.45 (m, 2H) , 7.26 (bs, IH) , 6.90 (d, IH), 6.85 (bs, IH), 3.75 (d, 2H) , 3.37 (s, 2H), 3.33 (s, 3H), 1.72 ( , 6H) , 1.16 (m, 5H) . Mass Spectrum (Chemical Ionization, 1% NH3 in CH4) m/z 304 (M+H) .
Step 8; (+/-)-2- (5- (l-Amino-ethyl) -2- cyclohexvlmethoxyphenyD -acetamide
This compound was synthesized in a manner similar to that described for Example 51, Step 4. The product was obtained as a solid. 1H NMR (400 MHz, DMSO): δ
7.11 (m,2H), 6.88 (m, IH) , 3.90 (q, IH), 3.71 (d, 2H) , 3.37 (s, 2H), 1.72 (m, 6H) , 1.20 (m, 8H) . Mass Spectrum (Chemical Ionization, 1% NH3 in CH4) m/z 290 (M+H) . Step 9: (S) -Phosphoric acid mono- (4- f2-acetvlamino-
2-r(RS)-l-(3-carbamoylmethy1-4-cyclohexylmethoxv- henvl)-ethylcarbamoy11 -ethyl) -phenyl) ester
The title compound was synthesized in a manner similar to that described for Example 44. The product was obtained as a colorless powder (93 mg) . HPLC 100%, rt = 17.8 and 18.0 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 574 (M-H).
EXAMPLE 66 Phosphoric acid mono- (4- (S) -2-acetylamino-2- \ 3- carbamoyl-4- (2-cvclohexv) - (RS) -1-methvl-ethoxv) - benzylcarbamoyl) -ethvl) -phenyl) ester
Figure imgf000098_0001
The title compound was synthesized in a manner similar to that described in Example 44. The product was obtained as a colorless solid (101 mg) . HPLC 98%, rt = 18.9 minutes, C18, eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA, and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 574 (M-H) .
EXAMPLE 67 Phosphoric acid mono- (4-f (S) -2-acetylamino-2- r (RS) -1-
(3-carbamoyl-4-cyclohexyl ethoxy-phenyl)-2-phenyl- ethylcarbamoyl) -ethvl) - henyl) ester
Figure imgf000098_0002
The title compound was synthesized in a manner similar to that described for Example 65 starting at step 3. The product was obtained as a colorless powder (26 mg) . HPLC 100%, rt = 20.2 and 20.5 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 636 (M-H) .
EXAMPLE 68 Phosphoric acid mono- f4-( (S)-2-acetylamino-2-f (RS)-1- f3-carbamovl-4-(3,5-dimethyl-benzyloxy)-phenyl) - ethylcarbamoyl)-ethyl)-phenyl) ester
Figure imgf000099_0001
The title compound was synthesized in a manner similar to that described for Example 51. The product was obtained as a colorless powder (16 mg) . HPLC 90%, rt = 16.9 and 17.3 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 582 (M-H).
EXAMPLE 69 Phosphoric acid mono-(4-f (S)-2-acetylamino-2- f (RS)-1- (3-carbamoyl-4-cyclohexyl ethoxy-phenyl)- butylcarbamoyl) -ethyl)-phenyl) ester
Figure imgf000100_0001
The title compound was synthesized in a manner similar to that described for Example 65 starting at step 3. The product was obtained as a colorless powder (32 mg) . HPLC 100%, rt - 19.2 and 19.5 minutes, C18, column eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 588 (M-H) .
EXAMPLE 70 (S)-Phosphoric acid mono-(4- [2-acetylamino-2-(4- cvclohexylmethoxy3-hydroxycarbamoyl-benzylcarbamoyl)- ethyl1-phenyl] ester
Figure imgf000100_0002
The title compound was synthesized in a manner similar to that described in Example 44, except that t- butyl-hydroxylamine was used instead of ammonia. This protecting group was not removed until the final deprotection step to give the product as a colorless solid (11 mg) . HPLC 92%, rt = 19.6 minutes, C18, eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA, and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 562 (M-H) .
EXAMPLE 71
(S) -Phosphoric acid mono-f4- r2-acetylamino-2- (4- cyclohexvlmethoxv-3-hydrazinocarbonyl-benzylcarbamoyl)- ethyl) -phenyl) ester
Figure imgf000101_0001
The title compound was synthesized in a manner similar to that described in Example 44, except that Boc-hydrazine was used instead of ammonia. This protecting group was not removed until the final deprotection step to give the product as a colorless solid (263 mg) . HPLC 96%, rt = 14.4 minutes, C18, eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA, and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 561 (M-H) .
EXAMPLE 72 (S) -Phosphoric acid mono- (4- f2-acetylamino-2- T3- carbamoyl-4- (2-ethvl-benzyloxy) -benzvlcarbamoyl) - ethyl) -phenyl) ester
Figure imgf000102_0001
The title compound was synthesized in a manner similar to that described in Example 44. The product was obtained as a colorless solid (58 mg) . HPLC 100%, rt = 16.8 minutes, C18, eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA, and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 568 (M-H) .
EXAMPLE 73 (4-f (S)-2-Acetvlamino-2- r(RS) -1-(3-carbamoyl-4- cvclohexvlmethoxyphenyl) -ethvlcarbamoyl) -ethyl) phenyl)-phosphonic acid
Figure imgf000102_0002
The title compound was synthesized in a manner similar to that described for Example 51 and 53. The product was obtained as a colorless powder (5 mg) . HPLC rt = 12.3 and 12.5 minutes, C18, column eluting with a gradient of 0% to 100% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 20 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 544 (M-H), EXAMPLE 74 Phosphoric acid mono-f4- r3-(3-carbamoyl-4- cyclohexvlmethoxv-benzyl)-ureido) -phenyl) ester
Figure imgf000103_0001
Step 1: l-isocyanato-4-(phenvlmethoxy)benzene To amine (21.20 mmol, 5.0 g) in toluene (10 ml) at room temperature was added phosgene in toluene (1.93 M, 106 mmol, 54 mL) . After refluxing 12 hours, the mixture was evaporated to dryness under reduced pressure. The crude product was used directly in the next step without further purification or characterization.
Step 2; 5- T3-(4-Benzyloxyphenyl)-ureidomethvl) - 2-cyclQhexylmethpχy-benaamide
This compound was synthesized in a manner similar to that described in Example 7 (step 1) .
Step 3; 2-Cvclohexylmethoxy5- T3- (4-hydroxy¬ phenyl)-ureidomethvl) -benzamide This compound was synthesized in a manner similar to that described in Example 6 (step 2) .
Step 4:Phosphoric acid mono-f4- \3-(3-carbamoyl-4- cyclohexylmethoxybenzyl)-ureido) -phenyl) ester
The title compound was synthesized in a manner similar to that described for Example 1. The product was obtained as a white solid (49 mg) . HPLC 100%, rt = 17.6 minutes, C18, eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water +0.1% ammonium hydroxide) m/z 476 (M-H) . EXAMPLE 75 (+/-) -Phosphoric acid mono- (5- f3- fl- (3-carbamoyl-4- cyclohexvlmethoxyphenyl) -ethyl) -ureido) -naphthalen-2- yl) ester
Figure imgf000104_0001
The title compound was synthesized in a manner similar to that described for Example 74. The product was obtained as a white solid (38 mg) . HPLC 100%, rt = 19.2 minutes, C18, eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water +0.1% ammonium hydroxide) m/z 540 (M-H).
EXAMPLE 76
Phosphoric acid mono-f4- \ 3 - (3-carbamoyl-4- cyclohexyl ethoxybenzvl) -ureidomethyl) -phenyl) ester
Figure imgf000104_0002
The title compound was synthesized in a manner similar to that described in Example 35 only the amine hydrochloride produced in Example 44 step 5 was employed as a partner in the urea forming reaction. The product was obtained as a colorless solid (78 mg) . HPLC 98%, rt = 17.5 minutes, C18, eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA, and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 490 (M-H).
EXAMPLE 77 fS-(R*rR»)1- or rs-(R*,S* l-2f2-Pimethyl-prppionic acid f r(4-f2-Acetylamino-2-ri-(3-carbamoyl-4- cyclohexylmethoxyphenyl)-ethylcarbamoyl) -ethyl)- phenyl)-difluoro-methvl) hydroxy-phosphinoyloxymethyl ester
Figure imgf000105_0001
Isomer- 1
The title compound was synthesized in a manner similar to that described in Example 55. The product was obtained as a colorless solid (6 mg) . HPLC 100%, rt = 18.8 minutes, C18, eluting with a gradient of 0% to 66% acetonitrile containing 0.1% TFA, and water containing 0.1% TFA over 22 minutes. Electrospray Mass Spectrum (50/50 acetonitrile/water + 0.1% ammonium hydroxide) m/z 708 (M-H) .
Inhibition of 125I-Phosphopeptide Binding to Immobilized Src SH2
The binding affinities of compounds of the present invention to Src SH2 was determined using a competitive radiolabeled phosphopeptide displacement assay. Specifically, binding of 125I-labeled Glu-Pro-Gln-pTyr- Glu-Glu-Ile-Pro-Ile-Tyr-Leu or 125I-labeled Glu-Pro- Gln- (4-(difluorophosphonomethyl) )-Phe-Glu-Glu-Ile-Pro- Ile-Tyr-Leu to a glutathione-S-transferase(GST) -Src SH2 fusion protein was performed in 20 mM Tris (pH 7.5), 150 mM NaCl, 5 mM EDTA, and 0.1% NP-40. Assay additions to a Millipore filter plate (0.45 mM PVDF) resulted in Src SH2 fusion protein-glutathione sepharose bead complex, 2.8 DH 125I-phosphopeptide and 2% DMSO + test compound at different concentrations. Binding was performed at room temperature for 20 minutes while continuously inverting the plate. Unbound 125I-phosphopeptide was separated from SH2-bound radiolabeled peptide by vacuum filtration and washing two times with 100 μL per well of assay buffer. Results are expressed as IC50 values in Table 1 below. Inhibition of Activated PDGF Receptor Binding to 35S-SH2 Domains
The binding of compounds of the present invention have been determined using 35S-labeled GST SH2 protein constructs and their binding to an immobilized PDGF receptor kinase domain. Binding of 35S-X-SH2-GST (X = Src, Abl, Grb2, p85-N, p85-C, Syp-N, and PLCγ-C domains) to immobilized PDGF receptor kinase domain was performed in a Millipore filter plate (0.45 mM PDVF) in 20 mM Tris buffer (pH 7.5), 150 mM NaCl, 10 mM mgCl2, and 0.1% triton. The assay was conducted at room temperature for 30 minutes while continuously inverting the plate. The resultant SH2-GST-PDGF receptor kinase complex was separated from excess 35S-SH2-GST protein by vacuum filtration and the amount of bound SH2-GST was determined by scintillation counting. Results are expressed as IC50 values and/or % inhibition at specified concentrations in Table 2 below. Inhibition of 125I-Phosphopeptide Binding to
Immobili ed Abl SH2 The binding affinities of compounds of the present invention to Abl SH2 was determined using a competitive radiolabeled phosphopeptide displacement assay. Specifically, binding of 125I-labeled Glu-Pro-Gln- (4- (difluorophosphonomethyl) )-Phe-Glu-Glu-Ile-Pro-Ile-Tyr- Leu to a glutathione-S-transferase(GST)-Abl SH2 fusion protein was performed in 20 mM Tris (pH 7.5), 150 mM NaCl, 5 mM EDTA, and 0.1% NP-40. Assay additions to a Millipore filter plate (0.45 mM PVDF) resulted in Abl SH2 fusion protein-glutathione sepharose bead complex, 2.8 nM 1 5I-phosphopeptide and 2% DMSO + test compound at different concentrations. Binding was performed at room temperature for 20 minutes while continuously inverting the plate, unbound 125I-phosphopeptide was separated from SH2-bound radiolabeled peptide by vacuum filtration and washing two times with 100 μL per well of assay buffer. Results are expressed as IC50 values in Table 3 below.
Inhibition of DNA Synthesis as Monitored by 3H- Thymidine Uptake
DNA synthesis occurs when a cell is exposed to Platelet Derived Growth Factor (PDGF) . For PDGF- induced DNA synthesis to occur, c-SRC or another member of the SRC gene family is required. The ability of the SRC SH2 inhibitors of the present invention to block
PDGF-stimulated DNA synthesis can be assessed using the protocol set forth below.
Swiss 3T3 cells were grown in 12-well plates for 3 days to approximately 50% confluency. The growth media was removed and replaced with 1.0 mL/well of assay buffer (Dulbecco's Modified Eagle Medium containing 0.2% Bovine Serum Albumin), and incubated for 24 hours to arrest growth.
Test compounds (5 μL) were add at 100 times their final concentration. Methyl sulfoxide was used as a control and levels in all wells are kept under 1.0% of DMSO. Cells were then stimulated with growth factor (PDGFbb or Fetal Bovine Serum), and allowed to incubate another 24 hours.
During the final 2 hours of treatment, 5 uL of 0.1 μCi/μL 3H-thymidine was added to each well (final = 0.5 μCi/well)
After incubation the medium was removed by aspiration, and each well was washed twice with ice- cold phosphate buffered saline (0.5 mL/wash) . 0.5 mL ice-cold 5% trichloroacetic acid was added to each well and incubated on ice for a minimum 10 minutes. The trichloroacetic acid solution was removed and each well was washed twice with 0.5 mL ice-cold trichloroacetic acid, and then once with ice-cold water (0.5 mL) . 0.5 mL of 2% Sodium Dodecyl Sulfate was added to each well to solubilize the cells and allowed to incubate at room temperature for 10-15 minutes. The solubilized cells were transfered to 20 mL scintillation vials. Each well was washed with 0.5 mL of water and combined with the corresponding scintillation vial. 10 mL of
Ready-Gel scintillation cocktail was added to each vial and the vials were then counted in a scintillation counter. Results are shown as percent of 3H-thymidine uptake relative to control in Table 4 below.
TABLE 1. Src SH2 Binding Data
IC
Example 50' SRC SH2
Binding (μM)
1 2.1
2 2.4 3 2.3
4 6.7
5 5.0
6 1.8
7 2.0 8 5.5
9 6.2
10 -25
11 2.1
12 0.7 13 4.4
14 10.0
15 7.0
16 9.8
17 3.2 18 3.3
19 5.6
20 20
21 5.5
22 8.3 23 6.2
24 7.2
25 7.1
26 9.8
27 8.0 28 12.0 TABLE 1. Src SH2 Binding Data (cont'd)
IC
Example 50' SRC SH2
Binding (μM)
29 8.3
30 5.5
31 3.9
32 9.2
33 9.3
34 -100
35 7.0
36 -30
37 6.7
38 4.1
39 9.7
40 7.0
41 6.6
42 -30
43 -20
44 6.5
45 -50
46 12.5
47 -20
48 -30
49 8.5
50 2.5
51 1.0
52 6.2
53 12.1
54 86
55 >100
56 3.8
57 33 TABLE 1 Src SH2 Binding Data (cont'd)
Example IC50, SRC SH2 Binding (μM)
58 21
59 1.1
60 1.9
61 16
62 8.3
63a 0.3
63b 29
64a 0.6
64b 11
65 9.2
66 3.8
67 1.0
68 1.1
69 2.1
70 24
71 18
72 6.2
73 8.6
74 20
75 7.2
76 34
TABLE 2
10
15
20
Figure imgf000112_0001
TABLE 3. Abl SH2 Binding Data
IC50, Abl SH2
Example Binding (μM)
5 1.7
10 1.6 11 17
12 2.5
24 36
34 42
35 >100 54 43
55 >100
57 20
58 -60
59 4.5 60 9.7
61 -100
62 44 63a 2.0 63b 59 64a , 2.4
64b 14
65 24
66 11
67 6.0 68 5.7
69 7.4
70 16
72 28
73 26 74 >100
75 -60
76 >100 TABLE 4. 3H-Thymidine Uptake in Swiss 3T3 Cell
Example % Uptake Relative to Control 77 34% 63a 73% 55 61%

Claims

A compound having the formula
Figure imgf000115_0001
wherein
U is -CO-, -CS-, -SO-, or -SO2- Y is
, ,
Figure imgf000115_0002
Figure imgf000115_0003
, , or
Figure imgf000115_0005
Figure imgf000115_0004
;
Figure imgf000115_0006
X is , , or a bond;
Figure imgf000115_0007
Figure imgf000115_0008
R1 is hydrogen, RCONR'-, RR'NCONR''-, RSO2NR'-, RCSNR'-, RR'NCSNR''-, RR'NSO2NR'
ROCONR'-, or ;
Figure imgf000115_0009
R2 is hydrogen, alkyl, cycloalkyl-(CH2)n-, substituted alkyl, aryl-(CH2)n-,
heteroaryl-(CH2)n-, -(CH2)n-CO2H, substituted cycloalkyl-(CH2)n-, substituted aryl-(CH2)n-, or substituted heteroaryl-(CH2)n-;
R3 is hydrogen, alkyl, cycloalkyl-(CH2)n-, substituted alkyl, aryl-(CH2)n-,
heteroaryl-(CH2)n-, -(CH2)n-CO2H, substituted cycloalkyl-(CH2)n-, substituted aryl-(CH2)n-, or substituted heteroaryl-(CH2)n-;
R4 is hydrogen or alkyl;
R5 is -NRR', , or
Figure imgf000116_0002
;
Figure imgf000116_0001
R6 and R9 are independently -OPO3RcRd,
-CF2PO3RcRd, -CH2PO3RcRd, -PO3RcRd, -SO3Rc,
-OSO3Rc, -CH2SO3Rc, -SO2NH2, -OSO2NH2, or
-CH2SO2NH2;
R7 and R8 are independently hydrogen, alkyl, substituted alkyl, halogen, -OR, -NRR', -COCF3,
-(CH2)nCH2OH, -(CH2)nCO2H, -(CH2)nCHO,
-(CH2)nNRR', or -Q-CH2-(CH2)n-NRR';
R10 is -(CH2)nCO2H, hydrogen, alkyl, aryl, substituted alkyl, or -(CH2)n-substituted aryl;
Rc and Rd are independently -R, -CH2CH2Z,
-CH2CHZ2, -CH2CZ3, or ;
Figure imgf000116_0003
Q is -O-, -NH-, - S -, -CH2O-, -CH2NH-, or -CH2S - ;
Z is -Cl, -Br, or - F ;
R, R', and R'' are independently hydrogen, alkyl, cycloalkyl-(CH2)n-, aryl-(CH2)n-,
heteroaryl-(CH2)n-, substituted alkyl, substituted cycloalkyl-(CH2)n-, substituted aryl-(CH2)n-,
-(CH2)nCO2H, or substituted heteroaryl-(CH2)n-; and each n is independently 0 to 5 , or the pharmaceutically acceptable salts, amides, esters, or prodrugs thereof.
2. A compound according to Claim 1 wherein
U is
X is and
Figure imgf000117_0001
.
Y is
Figure imgf000117_0004
3. A compound according Claim 2 wherein
R1 is RCONR'-, -NRCONR'R''-' -NRSO2R', or ;
Figure imgf000117_0005
R2, R4, R7, and R8 are hydrogen;
R3 is -(CH2)nCO2H, alkyl, or
-(CH2)n-substituted aryl;
R 5 is -NRR', ; and
Figure imgf000117_0002
R6 is -OPO3RcRd or -CF2PO3RcRd. 4. A compound according to Claim 2 wherein
R1 is CH3CONH-;
R2, R4, R7, and R8 are hydrogen;
R3 is -CH2CH2CO2H;
R'
R5 is -NRR', -NCH(CH3) ; and
Figure imgf000117_0003
R6 is -OPO3RcRd, -CF2PO3RcRd, or -PO3RcRd, A compound according to Claim 1 wherein
U is
is or and
Figure imgf000118_0002
Figure imgf000118_0003
X is .
Figure imgf000118_0004
6. The compound of Claim 5 wherein
R3 and R10 are -(CH2)nCO2H;
R4 is hydrogen;
R 5 is -NRR', ;
Figure imgf000118_0001
R7 and R8 are hydrogen; and
R6 is -OPO3RcRd, -CF2PO3RcRd, or -PO3RcRd.
7. A method of inhibiting the binding of a protein containing an SH2 domain to a cognate
phosphorylated protein, the method comprising administering to a patient in need of SH2 inhibition an SH2 inhibiting amount of a compound of Claim 1.
8. The method of Claim 7 wherein the protein
containing the SH2 domain is Src, Fyn, Lck, Yes, Blk, Lyn, Fgr, Hck, Yrk, or Abl.
9. The method of Claim 7 wherein the protein
containing the SH2 domain is pp60c-src kinase.
10. The method of Claim 7 wherein the cognate
phosphorylated protein is PDGF receptor protein, EGF receptor protein, HER2/Neu receptor protein, fibroblast growth factor receptor protein, focal adhesion kinase protein, pl30 protein, or p68 protein.
11. The method of Claim 7 wherein the patient in need of SH2 inhibition has a proliferative disease, cancer, restenosis, osteoporosis, inflammation, allergies, or cardiovascular disease.
12. A pharmaceutical composition that comprises a
compound of Claim 1 and a pharmaceutically acceptable carrier.
13. A method of treating a patient having a
proliferative disease, the method comprising administering to the patient a therapeutically effective amount of a compound of Claim 1.
14. A method of treating a patient having cancer, the method comprising administering to the patient a therapeutically effective amount of a compound of Claim 1.
15. A method of treating a patient having restenosis, the method comprising administering to the patient a therapeutically effective amount of a compound of Claim 1.
16. A method of treating a patient having
osteoporosis, the method comprising administering to the patient a therapeutically effective amount of a compound of Claim 1.
17. A method of treating a patient having
inflammation, the method comprising administering to the patient a therapeutically effective amount of a compound of Claim 1.
18. A method of treating a patient having allergies, the method comprising administering to the patient a therapeutically effective amount of a compound of Claim 1.
19. A method of treating a patient having
cardiovascular disease, the method comprising administering to the patient a therapeutically effective amount of a compound of Claim 1.
20. A compound of the formula
Figure imgf000120_0001
wherein U and W are independently -CO-, -CS-, -SO-, or -SO2-;
Y is
Figure imgf000121_0002
, , or
Figure imgf000121_0003
Figure imgf000121_0004
Figure imgf000121_0001
X is , , or a bond;
Figure imgf000121_0005
Figure imgf000121_0006
R11 is hydrogen, alkyl, -OH, substituted alkyl, or -NH2;
R12 is hydrogen or alkyl;
R13 is -(CH2)nCO2H, alkyl, -(CH2)n-aryl, -(CH2)n-heteroaryl, -(CH2)n-cycloalkyl, hydrogen, substituted cycloalkyl-(CH2)n _, substituted aryl-(CH2)n-, substituted heteroaryl-(CH2)n-, or substituted alkyl;
R14 and R17 are independently -OPO3RcRd, -CF2PO3RcRd, -CH2PO3RcRd, -PO3RcRd, -SO3Rc,
-OSO3Rc, -CH2SO3Rc, -SO2NH2, -OSO2NH2, or
-CH2SO2NH2;
R15 and R16 are independently hydrogen, alkyl, halogen, -OR, -NRR', -COCF3, -(CH2)nCH2OH, -(CH2)nCO2H, -(CH2)nNRR', -(CH2)nCHO, or
-Q-CH2-(CH2)n-NRR'; R18 is -(CH2)nCO2R, hydrogen, alkyl,
(CH2)nCONRR' , substituted alkyl, or
(CH2)n-substituted aryl;
R19 is hydrogen, RCONR'-, RR'NCONR''-,
RSO2NR'-, RR'NSO2NR' or ;
Figure imgf000122_0001
R20 is hydrogen, alkyl, cycloalkyl-(CH2)n-, substituted alkyl, aryl-(CH2)n-,
heteroaryl-(CH2)n-, -(CH2)n-CO2H, substituted cycloalkyl-(CH2)n-, substituted aryl-(CH2)n-, or substituted heteroaryl-(CH2)n-;
Ra is hydrogen, halogen, or alkyl;
Rb is hydrogen, alkyl, -OR, -O(CH2)n-aryl,
-NRR', -O(CH2)n-substituted alkyl, -SR,
-O(CH2)n-substituted aryl, or -O(CH2)n-cycloalkyl;
Rc and Rd are independently - R, -(CH2CH2Z, -CH2CHZ2, -CH2CZ3, or ;
Figure imgf000122_0002
Q is -O-, - NH-, - S -, -CH2O-, -CH2NH -, or -CH2S - ;
Z is -Cl, -Br, or -F;
R, R' , and R' ' are independently hydrogen, alkyl, cycloalkyl- (CH2)n-, aryl-(CH2)n-,
heteroaryl- (CH2)n-, -CH2-(CH2)n-CO2H, substituted cycloalkyl-(CH2)n-, substituted alkyl, substituted aryl-(CH2)n _, or substituted heteroaryl-(CH2)n-; and
each n is independently 0 to 5 , or the pharmaceutically acceptable salts, amides, esters, or prodrugs thereof.
21. A compound according to Claim 20 wherein Y is ;
Figure imgf000123_0001
U and W are -CO-; and
X is a bond.
22. A compound according to Claim 21 wherein
R13, Ra, R15, and R16 are hydrogen; R12 and R11 are hydrogen or alkyl;
Rb is -OR, -O(CH2)n-aryl, -O(CH2)nsubstituted aryl, or -O(CH2)ncycloalkyl; and
R14 is -OPO2RcRd or -CF2PO3RcRd.
23. A compound according to Claim 20 wherein
X is ;
Figure imgf000123_0002
Y is ; and
Figure imgf000123_0003
U and W are -CO-
24. A compound according to Claim 23 wherein
R19 is RCONR'- or RR'NCONR''-;
R20, R15, R13, R11, Ra, and R16 are hydrogen; R12 is alkyl or hydrogen;
Rb is -OR, -O(CH2)n-aryl, -O(CH2)nsubstituted aryl, or -O(CH2)ncycloalkyl; and
R14 is -OPO3RcRd or -CF2PO3RcRd.
25. A method of inhibiting the binding of a protein containing an SH2 domain to a cognate
phosphorylated protein, the method comprising administering to a patient in need of SH2 inhibition an SH2 inhibiting amount of a compound of Claim 20.
26. The method of Claim 25 wherein the protein
containing the SH2 domain is Src, Fyn, Lck, Yes, Blk, Lyn, Fgr, Hck, Yrk, or Abl.
27. The method of Claim 25 wherein the protein
containing the SH2 domain is pp60c-src kinase.
28. The method of Claim 25 wherein the cognate
phosphorylated protein is PDGF receptor protein, EGF receptor protein, HER2/Neu receptor protein, fibroblast growth factor receptor protein, focal adhesion kinase protein, pl30 protein, or p68 protein.
29. The method of Claim 25 wherein the patient in need of SH2 inhibition has a proliferative disease, cancer, restenosis, osteoporosis, inflammation, allergies, or cardiovascular disease.
30. A pharmaceutical composition that comprises a
compound of Claim 20 and a pharmaceutically acceptable carrier.
31. A method of treating a patient having a
proliferative disease, the method comprising administering to the patient a therapeutically effective amount of a compound of Claim 20.
32. A method of treating a patient having cancer, the method comprising administering to the patient a therapeutically effective amount of a compound of Claim 20.
33. A method of treating a patient having restenosis, the method comprising administering to the patient a therapeutically effective amount of a compound of Claim 20.
34. A method of treating a patient having
osteoporosis, the method comprising administering to the patient a therapeutically effective amount of a compound of Claim 20.
35. A method of treating a patient having
inflammation, the method comprising administering to the patient a therapeutically effective amount of a compound of Claim 20.
36. A method of treating a patient having allergies, the method comprising administering to the patient a therapeutically effective amount of a compound of Claim 20.
37. A method of treating a patient having
cardiovascular disease, the method comprising administering to the patient a therapeutically effective amount of a compound of Claim 20.
38. The compound that is:
[S-(R*,R*)]-4-[2-Acetylamino-3-(4- phosphonooxy-phenyl)-propionylamino]-4-[(3- cyclohexyl-propyl)-methyl-carbamoyl]-butyric acid;
4-[(S)-2-Acetylamino-3-(4-phosphonooxy- phenyl)-(S)-propionylamino]-4-(2-cyclohexyl-(S)-1- methyl-ethylcarbamoyl)-butyric acid;
4-[(S)-2-Acetylamino-3-(4-phosphonooxy- phenyl)-(S)-propionyl-amino]-4-(2-cyclohexyl-(S)- 1-methyl-ethyl)-methyl-carbamoyl)-butyric acid; 4-{(RS)-2-Acetylamino-3-[4-(difluorophosphono-methyl)-phenyl]-(S)-propionylamino)-4-[(3-cyclohexyl-propyl)-methyl-carbamoyl]-butyric acid;
[S-(R*,R*)]-4-{2-Acetylamino-3-[4-(difluoro-phosphono-methyl)-phenyl]-propionylamino)-4-[(3-cyclohexyl-propyl)-methyl-carbamoyl]-butyric acid;
[S-(R*,R*)]-4-[(3-Cyclohexyl-propyl)-methyl)-carbamoyl]-4-[2-[(morpholine-4-carbonyl)-amino]-3-(4-phosphonooxy-phenyl)-propionylamino]-butyric acid;
[S-(R*,R*)]-4-[(3-Cyclohexyl-propyl)-methyl-carbamoyl]-4-[2-(3-methyl-ureido)-3-(4-phosphonooxy-phenyl)-propionylamino]-butyric acid;
[S-(R*,R*)]-Phosphoric acid mono-[4-(2-acetylamino-2-{1-[(3-cyclohexyl-propyl)-methyl-carbamoyl]-propylcarbamoyl]-ethyl)-phenyl] ester;
[S-(R*,R*)]-Phosphoric acid mono-[4-(2-acetylamino-2-{1-[(3-cyclohexyl-propyl)-methyl-carbamoyl]-pentylcarbamoyl)-ethyl)-phenyl] ester;
[S-(R*,R*)]-{[4-(2-Acetylamino-2-{1-[(3-cyclohexyl-propyl)-methyl-carbamoyl]-propylcarbamoyl)-ethyl)-phenyl]-difluoro-methyl}-phosphonic acid;
4-[(S)-2-Acetylamino-3-(4-phosphonooxy-phenyl)-(S)-propionylamino]-5-oxo-5-((S)-2-phenethyl-pyrrolidin-1-yl)-pentanoic acid;
4-[(S)-2-Acetylamino-3-(4-phosphonooxy-phenyl)-(S)-propionylamino]-5-[2-((S)-2-cyclohexyl-ethyl)-pyrrolidin-1-yl]-5-oxo-pentanoic acid;
[S-(R*,R*)]-4-[2-Acetylamino-3-(4-phosphonooxy-phenyl)-propionylamino]-4-[2- (5,6,7,8-tetrahydro-naphthalen-1-yl)-ethylcarbamoyl]-butyric acid; [S-(R*,R*)]-4-[2-Acetylamino-3-(4-phosphonooxy-phenyl)-propionylamino]-4-(3-phenyl-propylcarbamoyl)-butyric acid;
[S-(R*,R*)]-4-[2-Acetylamino-3-(4-phosphonooxy-phenyl)-propionylamino]-4- [(naphthalen-1-ylmethyl)-carbamoyl]-butyric acid;
[S-(R*,R*)]-4-[2-Acetylamino-3-(4-phosphonooxy-phenyl)-propionylamino]-4-phenethylcarbamoyl-butyric acid;
4-[(S)-2-Acetylamino-3-(4-phosphonooxy-phenyl)-(S)-propionylamino]-4-[methyl-((S)-1-methyl-2-phenyl-ethyl)-carbamoyl]-butyric acid;
4-[(S)-2-Acetylamino-3-(4-phosphonooxy-phenyl)-(S)-propionylamino]-4-((S)-1-methyl-2-phenyl-ethylcarbamoyl)-butyric acid;
[S-(R*,R*)]-4-[2-Acetylamino-3-(4-phosphonooxy-phenyl)-propionylamino]-4-[methyl-(3-phenyl-propyl)-carbamoyl]-butyric acid;
[S-(R*,R*)]-Phosphoric acid mono-(4-(2-acetylamino-2-[2-(4-hydroxyphenyl)-1-(3-phenyl-propylcarbamoyl)-ethyl-carbamoyl]-ethyl)-phenyl) ester;
[S-(R*,R*)]-4-[2-Acetylamino-3-(4-phosphonooxy-phenyl)-propionylamino]-4-[2-(2-methoxyphenyl)-ethylcarbamoyl]-butyric acid;
[S-(R*,R*)]-4-[2-Acetylamino-3-(4-phosphonooxy-phenyl)-propionylamino]-4-(2-p-tolyl-ethylcarbamoyl)-butyric acid;
[S-(R*,R*)]-4-[2-Acetylamino-3-(4-phosphonooxy-phenyl)-propionylamino]-4-{[2-(2-methoxyphenyl)-ethyl]-methyl-carbamoyl)-butyric acid;
[S-(R*,R*)]-4-[2-Acetylamino-3-(4-phosphonooxyphenyl)-propionylamino]-4-[3-(3-carbamoylmethoxy-phenyl)-propylcarbamoyl]-butyric acid; [S-(R*,R*)]-4-[{2-[2-Acetylamino-3-(4-phosphonooxy-phenyl)-propionylamino]-propionyl)-(3-cyclohexyl-propyl)-amino]-butyric acid;
[S-(R*,R*)]-4-[{2-[2-Acetylamino-3-(4-phosphonooxyphenyl)-propionylamino]-propionyl)-(3-phenyl-propyl)-amino]-butyric acid;
[S-(R*,R*)]-5-[{2-[2-Acetylamino-3-(4-phosphonooxy-phenyl)-propionylamino]-propionyl)-(3-phenyl-propyl)-amino]-pentanoic acid;
[S-(R*,R*)]-Phosphoric acid mono-[4-(2-acetylamino-2-{1-[butyl-(3-phenyl-propyl)-carbamoyl]-ethylcarbamoyl)-ethyl)-phenyl] ester;
[S-(R*,R*)]-4-[2-Acetylamino-3-(4-phosphonooxy-phenyl)-propionylamino]-4-[3-(4-carboxymethoxyphenyl)-propylcarbamoyl]-butyric acid;
4-[(S)-2-Acetylamino-3-(4-phosphonooxy-phenyl)-(S)-[propionylamino]-5-((S)-2-benzyl-pyrrolidin-1-yl)-5-oxo-pentanoic acid;
L-Tyrosinamide, N-acetyl-O-phosphono-L-tyrosyl-N-(3-phenylpropyl)-O-phosphono-;
4-[(RS)-2-(Acetyl-methyl-amino)-3-(4-phosphonooxyphenyl)-(S)-propionylamino]-4-(3-phenyl-propylcarbamoyl)-butyric acid;
[S-(R*,R*)]-4-[2-Methanesulfonylamino-3-(4-phosphonooxyphenyl)-propionylamino]-4-(3-phenyl-propylcarbamoyl)-butyric acid;
[S-(R*,R*)]-[4-(2-Acetylamino-2-{1-[(3-cyclohexyl-propyl)-methyl-carbamoyl]-propylcarbamoyl)-ethyl)-phenyl]-phosphonic acid;
(S)-4-[3-Carboxymethyl-3-(4-phosphonooxy-benzyl)-ureido]-4-[(3-cyclohexyl-propyl)-methyl-carbamoyl]-butyric acid;
(S)-4-[(3-Cyclohexyl-propyl)-methyl¬carbamoyl]-4-[3-(6-phosphonoxynaphthalen-1-yl)-ureido]-butyric acid; [S-(R*,R*)]-4-[2-Acetylamino-3-(4-phosphonooxyphenyl)-propionylamino]-4-(5-methyl-hexylcarbamoyl)-butyric acid;
[S-(R*,R*)]-4-[2-Acetylamino-3-(4-phosphonooxyphenyl)-propionylamino]-4-(4-methyl-hexylcarbamoyl)-butyric acid;
Phosphoric acid mono-[4-(4-benzyloxy-3-carbamoyl-benzylcarbamoyl)-phenyl] ester;
Phosphoric acid mono-{4-[3-carbamoyl-4-(3-methyl-benzyloxy)-benzylcarbamoyl]-phenyl) ester;
Phosphoric acid mono-{4-[3-carbamoyl-4-(3,5-dimethyl-benzyloxy)-benzylcarbamoyl]-phenyl) ester;
Phosphoric acid mono-[4-[3-carbamoyl-4-(4-methyl-pentyloxy)-benzylcarbamoyl]-phenyl) ester;
Phosphoric acid mono-{4-[3-carbamoyl-4-(4-methyl-hexyloxy)-benzylcarbamoyl]-phenyl) ester;
Phosphoric acid mono-[4-(3-carbamoyl-4-cyclohexyl-methoxy-benzylcarbamoyl)-phenyl] ester;
[4-(3-Carbamoyl-4-cyclohexyImethoxy-benzylcarbamoyl)-phenyl]-difluoro-methyl-phosphonic acid;
Phosphoric acid mono-[4-(3-carbamoyl-4-cyclohexyImethoxybenzyIcarbamoyl)-2,6-dimethyl-phenyl] ester;
Phosphoric acid mono-[4-(3-carbamoyl-4-cyclohexyImethoxybenzylcarbamoyl)-2-chloro-phenyl] ester;
Phosphoric acid mono-[4-(4-cyclohexylmethoxy3-methylcarbamoyl-benzyIcarbamoyl)-phenyl] ester;
(S)-Phosphoric acid mono-{4-[2-acetylamino-2-(3-carbamoyl-4-cyclohexyImethoxy-benzylcarbamoyl)-ethyl]-phenyl) ester;
(S)-Phosphoric acid mono-(4-[2-acetylamino-2-[3-carbamoyl-4-(3,5-dimethyl-benzyloxy)-benzylcarbamoyl]-ethyl)-phenyl) ester; Phosphoric acid mono-(4-{(S)-2-acetylamino-2- [(RS)-1-(3-carbamoyl-4-cyclohexyImethoxyphenyl)- ethylcarbamoyl]-ethyl}-phenyl) ester;
(S)-Phosphoric acid mono-{4-[2-acetylureido- 2-(3-carbamoyl-4-cyclohexyImethoxy- benzylcarbamoyl)-ethyl]-phenyl) ester; and
(S)-{4-[2-Acetylamino-2-(3-carbamoyl-4- cyclohexyl-methoxy-benzylcarbamoyl)-ethyl]- phenyl)-difluoro-methyl-phosphonic acid.
39. A compound according Claim 2 wherein
R1 is RCONR'-, -NRCONR'R' '-, or -NRSO2R'; R2, R4, R7, and R8 are hydrogen;
R3 is -(CH2)nCO2H or alkyl; is -NRR',
Figure imgf000130_0001
, or ; and
Figure imgf000130_0002
R6 is - OPO3RcRd, -CF2PO3RcRd, or PO3RcRd
40. A compound according to Claim 1 wherein
U is -CO-;
X is a bond; and
is .
Figure imgf000130_0003
41. A compound according to Claim 40 wherein
R4, R7, and R8 are hydrogen;
R3 is -CH2CH2CO2H, or alkyl;
R5 is -NRR',
Figure imgf000130_0004
, or ; and
Figure imgf000130_0005
R6 is -OPO3RcRd, -CF2PO3RcRd, or -PO3RcRd.
42. A compound of the formula
Figure imgf000131_0001
wherein U and W are independently -CO-, -CS- -SO-, or -SO2-;
Y is
Figure imgf000131_0002
, , or
Figure imgf000131_0004
Figure imgf000131_0003
;
Figure imgf000131_0007
X is , , or a bond;
Figure imgf000131_0005
Figure imgf000131_0006
R11 is hydrogen, alkyl, -OH, substituted alkyl, or -NH2;
R12 is hydrogen or alkyl;
R13 is -(CH2)nCO2H, alkyl, - (CH2)n-aryl, -(CH2)n-heteroaryl, -(CH2)n-cycloalkyl, hydrogen, substituted cycloalkyl-(CH2)n-, substituted aryl- (CH2)n- , substituted heteroaryl- (CH2)n _ , or substituted alkyl;
R14 and R17 are independently -OPO3RcRd, -CF2PO3RcRd, -CH2PO3RcRd, -PO3RcRd, -SO3Rc,
-OSO3Rc, -CH2SO3Rc, -SO2NH2, -OSO2NH2, or
-CH2SO2NH2;
R15 and R16 are independently hydrogen, alkyl, halogen, -OR, -NRR', -COCF3, -(CH2)nCH2OH, -(CH2)nCO2H, -(CH2)nNRR', -(CH2)nCHO, or
-Q-CH2-(CH2)n-NRR' ;
R18 is -(CH2)nCO2R, hydrogen, alkyl,
-(CH2)nCONRR' , substituted alkyl, or
-(CH2)n-substituted aryl;
R19 is hydrogen, RCONR'-, RR'NCONR"-, RSO2NR'-, RR'NSO2NR''-, ROCONR'-, or ;
Figure imgf000132_0001
R20 is hydrogen, alkyl, cycloalkyl-(CH2)n-, substituted alkyl, aryl-(CH2)n-,
heteroaryl-(CH2)n-, -(CH2)n-CO2H, substituted cycloalkyl-(CH2)n-, substituted aryl-(CH2)n-, or substituted heteroaryl-(CH2)n-;
Ra is hydrogen, halogen, or alkyl;
Rb is hydrogen, alkyl, -OR, -O(CRR')n-aryl,
-NRR', -O(CRR')n-substituted alkyl, -SR,
-O(CRR')n-substituted aryl, or -O(CRR')n- cycloalkyl;
Rc and Rd are independently -R, -CH2CH2Z,
-CH2CHZ2, -CH2CZ3, or ;
Figure imgf000132_0002
Q is -O-, -NH-, -S-, -CH2O-, -CH2NH-, or -CH2S-;
Z is -Cl, -Br, or -F; R, R', and R'' are each independently
hydrogen, alkyl, cycloalkyl-(CH2)n-, aryl-(CH2)n-, heteroaryl-(CH2)n-, -CH2-(CH2)n-CO2H, substituted cycloalkyl-(CH2)n-, substituted alkyl, substituted aryl-(CH2)n-, or substituted heteroaryl-(CH2)n-; and
each n is independently 0 to 5, or the pharmaceutically acceptable salts, amides, esters, or prodrugs thereof.
43. A compound according to Claim 42 wherein X is ;
Figure imgf000133_0001
Y is ; and
Figure imgf000133_0002
U and W are -CO-
44. A compound according to Claim 43 wherein
R19 is RCONR'- or RR'NCONR''-;
R20, R15, R13, R11, Ra, and R16 are hydrogen; R12 is alkyl or hydrogen;
Rb is -OR, -O(CRR' )n-aryl, -O(CRR')n substituted aryl, or -O(CRR')ncycloalkyl; and
R14 is -OPO3RcRd, -CF2PO3RcRd, or -PO3RcRd.
45. A compound according to Claim 42 wherein
X is ;
Figure imgf000133_0003
Y is ; and
Figure imgf000133_0004
U and W are -CO- .
46. A compound according to Claim 42 wherein
R15, R13, R11, Ra, and R16 are hydrogen;
R12 is alkyl or hydrogen;
Rb is -OR, -O(CRR')n-aryl, -O(CRR')n substituted aryl, or -O(CRR' )ncycloalkyl; and
R14 is -OPO3RcRd, -CF2PO3RcRd, or -PO3RcR
47. A compound according to Claim 42 wherein X is ;
Figure imgf000134_0001
Y is ; and
Figure imgf000134_0002
U and W are -CO-.
48. A compound according to Claim 47 wherein
R13, R11, and Ra are hydrogen;
R12 is alkyl or hydrogen;
Rb is -OR, -O(CRR' )n-aryl, -O(CRR')n substituted aryl, or -O(CRR' )ncycloalkyl; and
R17 is -OPO3 RcRd, -CF2PO3RcRd, or PO3RcRd.
49. The compound that is:
[S-(R*,R*)]-4-[2-Acetylamino-3-(4-phosphono- phenyl)-propionylamino]-4-[(3-cyclohexyl-propyl)- methyl-carbamoyl]-butyric acid;
[S-(R*,R*)]-2,2-Dimethyl-propionic acid {[4- (2-acetylamino-2-[1-[(3-cyclohexyl-propyl)-methyl- carbamoyl]-propylcarbamoyl)-ethyl)-phenyl]- difluoro-methyl } -hydroxyphosphinoyloxymethyl ester;
[ S- ( R*,R* ) ] -4 - [ 2-Acetylamino- 3 - ( 4 -phosphonooxyphenyl)-propionylamino]-4-(2-adamantan-1-yl-ethylcarbamoyl)-butyric acid;
(S)-Phosphoric acid mono-(4-{2-acetylamino-2- [1-ethyl-1-(3-phenyl-propylcarbamoyl)-propylcarbamoyl]-ethyl)-phenyl) ester;
[S-(R*,R*)]-Phosphoric acid mono-(4-{2-acetylamino-2- [2-hydroxyl- ( 3 -phenyl-propylcarbamoyl)-propylcarbamoyl]-ethyl]-phenyl) ester;
[S-(R*,R*)]-4-[(3-Cyclohexyl-propyl)-methyl-carbamoyl]-4-[2-(3,3-dimethyl-ureido)-3-(4-phosphonooxyphenyl)-propionylamino]-butyric acid;
5- [(3-cyclohexylpropyl)methylamino]-4-[[2- [[(dimethylamino)-sulfonyl]amino]-1-oxo-3-[4-(phosphonooxy)phenyl]propyl]amino]-5-oxo-pentanoic acid;
(S)-4-[(3-Cyclohexyl-propyl)-methyl-carbamoyl]-4-[3-(4-phosphonooxy-benzyl)-ureido]-butyric acid;
(S)-4-[(3-Cyclohexyl-propyl)-methyl-carbamoyl]-4-(4-phosphonooxybenzoylamino)-butyric acid;
[S-(R*,R*)]- or [S-(R*,S*)]-[(4-{2- Acetylamino-2-[1-(3-carbamoyl-4-cyclohexyImethoxy-phenyl)-ethylcarbamoyl]-ethyl)-phenyl)-difluoro-methyl]-phosphonic acid;
[S-(R*,R*)]- or [S-(R*,S*)]-[(4-{2- Acetylamino-2-[1-(3-carbamoyl-4-cyclohexyImethoxy-phenyl)-ethylcarbamoyl]-ethyl)-phenyl)-difluoro- methyl]-phosphonic acid;
Phosphoric acid mono-[4-(2-acetylamino-2-{1- [3-carbamoyl-4-(2-cyclohexy1-1-methyl-ethoxy)-phenyl]-ethylcarbamoyl)-ethyl)-phenyl] ester; Phosphoric acid mono-[4-(2-acetylamino-2-{1-[3-carbamoyl-4-(2-cyclohexyl-1-methyl-ethoxy)-phenyl]-ethylcarbamoyl)-ethyl)-phenyl] ester;
(S)-Phosphoric acid mono-(4-{2-acetylamino-2-[(RS)-1-(3-carbamoylmethyl-4-cyclohexyImethoxy-phenyl)-ethylcarbamoyl]-ethyl]-phenyl) ester;
Phosphoric acid mono-(4-{(S)-2-acetylamino-2- [3-carbamoyl-4-(2-cyclohexyl-(RS)-1-methyl-ethoxy)-benzylcarbamoyl]-ethyl)-phenyl) ester;
Phosphoric acid mono-(4-{(S)-2-acetylamino-2-[(RS)-1-(3-carbamoyl-4-cyclohexyImethoxy-phenyl)-2-phenyl-ethylcarbamoyl]-ethyl)-phenyl) ester;
Phosphoric acid mono-[4-((S)-2-acetylamino-2-{(RS)-1-[3-carbamoyl-4-(3,5-dimethyl-benzyloxy)-phenyl]-ethylcarbamoyl)-ethyl)-phenyl] ester;
Phosphoric acid mono-(4-{(S)-2-acetylamino-2-[(RS)-1-(3-carbamoy1-4-cyclohexyImethoxyphenyl)-butylcarbamoyl]-ethyl)-phenyl) ester;
(S)-Phosphoric acid mono-{4-[2-acetylamino-2-(4-cyclohexyImethoxy-3-hydroxycarbamoyl-benzylcarbamoyl)-ethyl]-phenyl) ester;
(S)-Phosphoric acid mono-{4-[2-acetylamino-2- (4-cyclohexyImethoxy3-hydrazinocarbonyl-benzylcarbamoyl)-ethyl]-phenyl) ester;
(S)-Phosphoric acid mono-(4-{2-acetylamino-2- [3-carbamoyl-4-(2-ethyl-benzyloxy)-benzylcarbamoyl]-ethyl)-phenyl) ester;
(4-{(S)-2-Acetylamino-2-[(RS)-1-(3-carbamoyl-4-cyclohexyImethoxyphenyl)-ethylcarbamoyl]-ethyl)-phenyl)-phosphonic acid;
Phosphoric acid mono-{4-[3-(3-carbamoyl-4-cyclohexyImethoxybenzyl)-ureido]-phenyl) ester;
(+/-)-Phosphoric acid mono-(5-[3-[1-(3-carbamoyl-4-cyclohexyImethoxy-phenyl)-ethyl]-ureido)-naphthalen-2-yl) ester; Phosphoric acid mono-{4-[3-(3-carbamoy1-4-cyclohexyImethoxy-benzyl)-ureidomethyl]-phenyl)ester; and
[S-(R*,R*)]- or [S-(R*,S*)]-2,2-Dimethyl-propionic acid {[(4-[2-Acetylamino-2-[1-(3-carbamoyl-4-cyclohexyImethoxyphenyl)-ethylcarbamoyl]-ethyl)-phenyl)-difluoro-methyl) hydroxy-phosphinoyloxymethyl ester.
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