US20060159627A1 - Method of preventing and treating airway remodeling and pulmonary inflammation using A2B adenosine receptor antagonists - Google Patents

Method of preventing and treating airway remodeling and pulmonary inflammation using A2B adenosine receptor antagonists Download PDF

Info

Publication number
US20060159627A1
US20060159627A1 US11/251,450 US25145005A US2006159627A1 US 20060159627 A1 US20060159627 A1 US 20060159627A1 US 25145005 A US25145005 A US 25145005A US 2006159627 A1 US2006159627 A1 US 2006159627A1
Authority
US
United States
Prior art keywords
optionally substituted
propyl
dione
methyl
pyrazol
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Abandoned
Application number
US11/251,450
Other languages
English (en)
Inventor
Dewan Zeng
Michael Blackburn
Luiz Belardinelli
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Gilead Sciences Inc
Original Assignee
CV Therapeutics Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by CV Therapeutics Inc filed Critical CV Therapeutics Inc
Priority to US11/251,450 priority Critical patent/US20060159627A1/en
Assigned to CV THERAPEUTICS, INC. reassignment CV THERAPEUTICS, INC. ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: BELARDINELLI, LUIZ, BLACKBUM, MICHAEL, ZENG, DEWAN
Publication of US20060159627A1 publication Critical patent/US20060159627A1/en
Assigned to CV THERAPEUTICS, INC. reassignment CV THERAPEUTICS, INC. ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: ZENG, DEWAN, BLACKBURN, MICHAEL R., BELARDINELLI, LUIZ
Priority to US12/605,783 priority patent/US8466129B2/en
Assigned to GILEAD PALO ALTO, INC. reassignment GILEAD PALO ALTO, INC. MERGER (SEE DOCUMENT FOR DETAILS). Assignors: APEX MERGER SUB, INC., CV THERAPEUTICS, INC.
Assigned to GILEAD SCIENCES, INC. reassignment GILEAD SCIENCES, INC. ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: GILEAD PALO ALTO, INC.
Abandoned legal-status Critical Current

Links

Images

Classifications

    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • A61K31/33Heterocyclic compounds
    • A61K31/395Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/519Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
    • A61K31/52Purines, e.g. adenine
    • A61K31/522Purines, e.g. adenine having oxo groups directly attached to the heterocyclic ring, e.g. hypoxanthine, guanine, acyclovir
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00Drugs for disorders of the respiratory system
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00Drugs for disorders of the respiratory system
    • A61P11/06Antiasthmatics
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00Drugs for disorders of the respiratory system
    • A61P11/08Bronchodilators
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00Antineoplastic agents
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00

Definitions

  • the present invention relates to methods of preventing airway remodeling and/or pulmonary inflammation using A 2B adenosine receptor antagonists.
  • This invention finds utility in the treatment and prevention of asthma, COPD, pulmonary fibrosis, emphysema, and other pulmonary diseases.
  • the invention also relates to pharmaceutical compositions for use in the method.
  • Airway remodeling typically takes the form of airway wall thickening, the development of subepithelial fibrosis, increased myocyte muscle mass, myofibroblast hyperplasia, and mucus metaplasia.
  • Airway remodeling is also a common factor in the progression of chronic obstructive pulmonary disorder (COPD), and pulmonary fibrosis.
  • COPD chronic obstructive pulmonary disorder
  • Pulmonary inflammation is also a common component in the development of airway remodeling and may be typified by bronchiolitis, alveolitis, and/or vasculitis.
  • Adenosine is known to play a role in asthma and COPD (See, Spicuzza et al. (2003) TiPS 24(8):409-4130; Mann et al, (1986) J Appl Physiol 61:1667-1676; and Feoktistov et al, (1998) Trends Pharmacol Sci 19:148-153.)
  • the clinical evidence supporting the involvement of adenosine includes:
  • Adenosine is a naturally occurring nucleoside, which exerts its biological effects by interacting with a family of adenosine receptors known as A 1 , A 2A , A 2B , and A 3 , all of which modulate important physiological processes.
  • a 2B adenosine receptors are believed to be most significantly involved in asthma via their connection to mast cell activation, vasodilation, and regulation of cell growth (See Adenosine A 2B Receptors as Therapeutic Targets, Drug Dev Res 45:198; Feoktistov et al., Trends Pharmacol Sci 19:148-153).
  • adenosine A 2B receptor antagonists have been shown to affect the activation of mast cell and have thus been implicated in the inhibition of the acute airway hyperresponsiveness. Surprisingly, it has now been found that A 2B adenosine receptor antagonists are also useful in the prevention of airway remodeling and pulmonary inflammation.
  • a 2B antagonists i.e., compounds that inhibit the A 2B adenosine receptor.
  • a method for the treatment and prevention of airway remodeling and/or pulmonary inflammation by administration of a therapeutically effective amount of an A 2B receptor antagonist to a mammal that is genetically and/or environmentally predisposed to airway remodeling and pulmonary inflammation.
  • compositions comprising a therapeutically effective amount of an A 2B receptor antagonist, and at least one pharmaceutically acceptable carrier.
  • the formulation is preferably for oral administration.
  • R 1 and R 2 are independently hydrogen, optionally substituted lower alkyl, or a group -D-E, in which D is a covalent bond or alkylene, and E is optionally substituted phenyl, optionally substituted cycloalkyl, optionally substituted alkenyl, or optionally substituted alkynyl, particularly those in which R 3 is hydrogen.
  • a first preferred class of compounds include those in which R 1 and R 2 are independently lower alkyl optionally substituted by cycloalkyl, preferably n-propyl, and X is optionally substituted phenylene.
  • R 1 and R 2 are independently lower alkyl optionally substituted by cycloalkyl, preferably n-propyl, and X is optionally substituted phenylene.
  • a preferred subclass of compounds are those in which Y is alkylene, including alkylene in which a carbon atom is replaced by oxygen, preferably —O—CH 2 —, more especially where the oxygen is the point of attachment to phenylene.
  • Z is optionally substituted oxadiazole, particularly optionally substituted [1,2,4]-oxadiazol-3-yl, especially [1,2,4]-oxadiazol-3-yl substituted by optionally substituted phenyl or optionally substituted pyridyl.
  • a second preferred class of compounds include those in which X is optionally substituted 1,4-pyrazolene.
  • a preferred subclass of compounds are those in which Y is a covalent bond or alkylene, especially lower alkylene, and Z is hydrogen, optionally substituted phenyl, optionally substituted pyridyl, or optionally substituted oxadiazole.
  • one preferred embodiment includes compounds in which R 1 is lower alkyl optionally substituted by cycloalkyl, and R 2 is hydrogen.
  • a more preferred embodiment includes those compounds in which Y is —(CH 2 )— or —CH(CH 3 )— and Z is optionally substituted phenyl, or Y is —(CH 2 )— or —CH(CH 3 )— and Z is optionally substituted oxadiazole, particularly 3,5-[1,2,4]-oxadiazole, or Y is —(CH 2 )— or —CH(CH 3 )— and Z is optionally substituted pyridyl.
  • R 1 and R 2 are independently lower alkyl optionally substituted by cycloalkyl, especially n-propyl.
  • Y is a covalent bond, —(CH 2 )— or —CH(CH 3 )— and Z is hydrogen, optionally substituted phenyl, or optionally substituted pyridyl, particularly where Y is a covalent bond and Z is hydrogen.
  • the preferred compounds for use in the invention include, but are not limited to:
  • FIG. 1 depicts the differences in pulmonary histopathology in adenosine deaminase (ADA) ⁇ / ⁇ mice following treatment with adenosine A 2B receptor antagonists as described in Example 21.
  • the lungs were collected and processed for histological analysis using H&E staining.
  • A Lung from ADA+ vehicle treated mouse.
  • B Lung from ADA ⁇ / ⁇ vehicle treated mouse.
  • C Lung from ADA ⁇ / ⁇ adenosine A 2B receptor antagonist treated mouse.
  • D Higher magnification section through the lung of an ADA+ vehicle treated mouse.
  • E Higher magnification section though the lung of an ADA ⁇ / ⁇ vehicle treated mouse.
  • FIG. 2 shows the effects of an adenosine A 2B receptor antagonist (A 2B ⁇ A) on airway cellularity as described in Example 21.
  • A Mice were lavalged with PBS and total cell counts determined.
  • FIG. 3 presents the result of BAL cellularity in ADA ⁇ / ⁇ mice treated with an adenosine A 2B receptor antagonist as described in Example 21.
  • BAL fluid was collected from the lungs of mice and cells were cytospun onto microscope slides and stained with DiffQuick.
  • A ADA+ vehicle treated
  • B ADA+ adenosine A 2B receptor antagonist treated
  • C ADA ⁇ / ⁇ vehicle treated
  • D ADA ⁇ / ⁇ adenosine A 2B receptor antagonist treated.
  • FIG. 4 illustrates transcript levels of various pro-inflammatory cytokines were measure in whole lung extracts using quantitative RT-PCR as described in Example 21.
  • A IL-6
  • B Eotaxin 1
  • FIG. 5 shows the results of ⁇ -smooth muscle actin immunostaining. Lung sections were stained with antiserum against ⁇ -smooth muscle actin to visualize myofibroblast (brown).
  • A Lung from an ADA + vehicle treated mouse.
  • B Lung from an ADA ⁇ / ⁇ vehicle treated mouse.
  • FIG. 6 depicts collagen deposition in the lungs of adenosine A 2B receptor antagonist-treated ADA ⁇ / ⁇ mice as described in Example 21. Lung sections were stained with Masson's Trichrome to visualize collagen deposition (blue).
  • A Lung from an ADA + vehicle treated mouse.
  • B Lung from an ADA + adenosine A 2B receptor antagonist treated mouse.
  • C Lung from an ADA ⁇ / ⁇ vehicle treated mouse.
  • E ⁇ -1 procollagen transcript levels are presented as mean pg transcripts/ ⁇ g RNA+SEM.
  • FIG. 7 shows the expression of fibrosis associated genes.
  • RNA was extracted from whole lungs for analysis using quantitative RT-PCR for various fibrosis associated transcripts. Results demonstrate that lungs from an adenosine A 2B receptor antagonist treated ADA ⁇ / ⁇ mice exhibit lower levels of transcripts for TGF- ⁇ 1 (A), osteopontin (OPN) (B), and plasminogen activator inhibitor-1 (Pai-1) (C), as compared to that seen in the lungs of vehicle treated ADA ⁇ / ⁇ mice.
  • A TGF- ⁇ 1
  • OPN osteopontin
  • Pai-1 plasminogen activator inhibitor-1
  • FIG. 8 shows the elevation of genes associated with alveolar airway destruction.
  • Transcript levels of TIMP-1, MMP-9, and MMP-12 were measured in whole lung RNA extracts using quantitative RT-PCR.
  • A TIMP-1.
  • B MMP-9.
  • C MMP-12.
  • FIG. 9 presents alveolar destruction in ADA ⁇ / ⁇ mice treated with an adenosine A 2B receptor antagonist as described in Example 21.
  • Lungs were collected and processed for histological analysis using H&E staining.
  • A Lung from ADA+ vehicle treated mouse
  • B Lung from ADA ⁇ / ⁇ vehicle treated mouse
  • C Lung from ADA ⁇ / ⁇ mouse treated with an adenosine A 2B receptor antagonist.
  • Photographs were all taken at the same magnification (10 ⁇ ) and represent findings from 6 different animals for each treatment group.
  • alkyl refers to a monoradical branched or unbranched saturated hydrocarbon chain having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 carbon atoms. This term is exemplified by groups such as methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, t-butyl, n-hexyl, n-decyl, tetradecyl, and the like.
  • substituted alkyl refers to:
  • lower alkyl refers to a monoradical branched or unbranched saturated hydrocarbon chain having 1, 2, 3, 4, 5, or 6 carbon atoms. This term is exemplified by groups such as methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, t-butyl, n-hexyl, and the like.
  • substituted lower alkyl refers to lower alkyl as defined above having 1 to 5 substituents, preferably 1, 2, or 3 substituents, as defined for substituted alkyl, or a lower alkyl group as defined above that is interrupted by 1, 2, 3, 4, or 5 atoms as defined for substituted alkyl, or a lower alkyl group as defined above that has both 1, 2, 3, 4 or 5 substituents as defined above and is also interrupted by 1, 2, 3, 4, or 5 atoms as defined above.
  • alkylene refers to a diradical of a branched or unbranched saturated hydrocarbon chain, having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 carbon atoms, preferably 1-10 carbon atoms, more preferably 1, 2, 3, 4, 5 or 6 carbon atoms.
  • This term is exemplified by groups such as methylene (—CH 2 —), ethylene (—CH 2 CH 2 —), the propylene isomers (e.g., —CH 2 CH 2 CH 2 — and —CH(CH 3 )CH 2 —) and the like.
  • lower alkylene refers to a diradical of a branched or unbranched saturated hydrocarbon chain, preferably having from 1, 2, 3, 4, 5, or 6 carbon atoms.
  • lower alkylene refers to a diradical of a branched or unbranched saturated hydrocarbon chain, preferably having from 1, 2, 3, 4, 5, or 6 carbon atoms.
  • substituted alkylene refers to:
  • aralkyl refers to an aryl group covalently linked to an alkylene group, where aryl and alkylene are defined herein.
  • Optionally substituted aralkyl refers to an optionally substituted aryl group covalently linked to an optionally substituted alkylene group.
  • Such aralkyl groups are exemplified by benzyl, phenylethyl, 3-(4-methoxyphenyl)propyl, and the like.
  • alkoxy refers to the group R—O—, where R is optionally substituted alkyl or optionally substituted cycloalkyl, or R is a group —Y-Z, in which Y is optionally substituted alkylene and Z is optionally substituted alkenyl, optionally substituted alkynyl; or optionally substituted cycloalkenyl, where alkyl, alkenyl, alkynyl, cycloalkyl and cycloalkenyl are as defined herein.
  • Preferred alkoxy groups are optionally substituted alkyl-O— and include, by way of example, methoxy, ethoxy, n-propoxy, iso-propoxy, n-butoxy, tert-butoxy, sec-butoxy, n-pentoxy, n-hexoxy, 1,2-dimethylbutoxy, trifluoromethoxy, and the like.
  • alkylthio refers to the group R—S—, where R is as defined for alkoxy.
  • alkenyl refers to a monoradical of a branched or unbranched unsaturated hydrocarbon group preferably having from 2 to 20 carbon atoms, more preferably 2 to 10 carbon atoms and even more preferably 2 to 6 carbon atoms and having 1-6, preferably 1, double bond (vinyl).
  • Preferred alkenyl groups include ethenyl or vinyl (—CH ⁇ CH 2 ), 1-propylene or allyl (—CH 2 CH ⁇ CH 2 ), isopropylene (—C(CH 3 ) ⁇ CH 2 ), bicyclo[2.2.1]heptene, and the like. In the event that alkenyl is attached to nitrogen, the double bond cannot be alpha to the nitrogen.
  • lower alkenyl refers to alkenyl as defined above having from 2 to 6 carbon atoms.
  • substituted alkenyl refers to an alkenyl group as defined above having 1, 2, 3, 4 or 5 substituents, and preferably 1, 2, or 3 substituents, selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkenyl, acyl, acylamino, acyloxy, amino, aminocarbonyl, alkoxycarbonylamino, azido, cyano, halogen, hydroxy, keto, thiocarbonyl, carboxy, carboxyalkyl, arylthio, heteroarylthio, heterocyclylthio, thiol, alkylthio, aryl, aryloxy, heteroaryl, aminosulfonyl, aminocarbonylamino, heteroaryloxy, heterocyclyl, heterocyclooxy, hydroxyamino, alkoxyamino, nitro, —SO-alkyl, —SO-aryl
  • substituents may optionally be further substituted by 1, 2, or 3 substituents chosen from alkyl, carboxy, carboxyalkyl, aminocarbonyl, hydroxy, alkoxy, halogen, CF 3 , amino, substituted amino, cyano, and —S(O) n R, where R is alkyl, aryl, or heteroaryl and n is 0, 1 or 2.
  • alkynyl refers to a monoradical of an unsaturated hydrocarbon, preferably having from 2 to 20 carbon atoms, more preferably 2 to 10 carbon atoms and even more preferably 2 to 6 carbon atoms and having at least 1 and preferably from 1-6 sites of acetylene (triple bond) unsaturation.
  • Preferred alkynyl groups include ethynyl, (—C—CH), propargyl (or prop-1-yn-3-yl, —CH 2 C ⁇ CH), and the like. In the event that alkynyl is attached to nitrogen, the triple bond cannot be alpha to the nitrogen.
  • substituted alkynyl refers to an alkynyl group as defined above having 1, 2, 3, 4 or 5 substituents, and preferably 1, 2, or 3 substituents, selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkenyl, acyl, acylamino, acyloxy, amino, aminocarbonyl, alkoxycarbonylamino, azido, cyano, halogen, hydroxy, keto, thiocarbonyl, carboxy, carboxyalkyl, arylthio, heteroarylthio, heterocyclylthio, thiol, alkylthio, aryl, aryloxy, heteroaryl, aminosulfonyl, aminocarbonylamino, heteroaryloxy, heterocyclyl, heterocyclooxy, hydroxyamino, alkoxyamino, nitro, —SO-alkyl, —SO-
  • substituents may optionally be further substituted by 1, 2, or 3 substituents chosen from alkyl, carboxy, carboxyalkyl, aminocarbonyl, hydroxy, alkoxy, halogen, CF 3 , amino, substituted amino, cyano, and —S(O) n R, where R is alkyl, aryl, or heteroaryl and n is 0, 1 or 2.
  • aminocarbonyl refers to the group —C(O)NRR where each R is independently hydrogen, alkyl, aryl, heteroaryl, heterocyclyl or where both R groups are joined to form a heterocyclic group (e.g., morpholino). Unless otherwise constrained by the definition, all substituents may optionally be further substituted by 1-3 substituents chosen from alkyl, carboxy, carboxyalkyl, aminocarbonyl, hydroxy, alkoxy, halogen, CF 3 , amino, substituted amino, cyano, and —S(O) n R, where R is alkyl, aryl, or heteroaryl and n is 0, 1 or 2.
  • acylamino refers to the group —NRC(O)R where each R is independently hydrogen, alkyl, aryl, heteroaryl, or heterocyclyl. Unless otherwise constrained by the definition, all substituents may optionally be further substituted by 1-3 substituents chosen from alkyl, carboxy, carboxyalkyl, aminocarbonyl, hydroxy, alkoxy, halogen, CF 3 , amino, substituted amino, cyano, and —S(O) n R, where R is alkyl, aryl, or heteroaryl and n is 0, 1 or 2.
  • acyloxy refers to the groups —O(O)C-alkyl, —O(O)C-cycloalkyl, —O(O)C-aryl, —O(O)C-heteroaryl, and —O(O)C-heterocyclyl. Unless otherwise constrained by the definition, all substituents may be optionally further substituted by alkyl, carboxy, carboxyalkyl, aminocarbonyl, hydroxy, alkoxy, halogen, CF 3 , amino, substituted amino, cyano, or —S(O) n R, where R is alkyl, aryl, or heteroaryl and n is 0, 1 or 2.
  • aryl refers to an aromatic carbocyclic group of 6 to 20 carbon atoms having a single ring (e.g., phenyl) or multiple rings (e.g., biphenyl), or multiple condensed (fused) rings (e.g., naphthyl or anthryl).
  • Preferred aryls include phenyl, naphthyl and the like.
  • arylene refers to a diradical of an aryl group as defined above. This term is exemplified by groups such as 1,4-phenylene, 1,3-phenylene, 1,2-phenylene, 1,4′-biphenylene, and the like.
  • such aryl or arylene groups can optionally be substituted with from 1 to 5 substituents, preferably 1 to 3 substituents, selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkenyl, acyl, acylamino, acyloxy, amino, aminocarbonyl, alkoxycarbonylamino, azido, cyano, halogen, hydroxy, keto, thiocarbonyl, carboxy, carboxyalkyl, arylthio, heteroarylthio, heterocyclylthio, thiol, alkylthio, aryl, aryloxy, heteroaryl, aminosulfonyl, aminocarbonylamino, heteroaryloxy, heterocyclyl, heterocyclooxy, hydroxyamino, alkoxyamino, nitro, —SO-alkyl,
  • substituents may optionally be further substituted by 1-3 substituents chosen from alkyl, carboxy, carboxyalkyl, aminocarbonyl, hydroxy, alkoxy, halogen, CF 3 , amino, substituted amino, cyano, and —S(O) n R, where R is alkyl, aryl, or heteroaryl and n is 0, 1 or 2.
  • aryloxy refers to the group aryl-O— wherein the aryl group is as defined above, and includes optionally substituted aryl groups as also defined above.
  • arylthio refers to the group R—S—, where R is as defined for aryl.
  • amino refers to the group —NH 2 .
  • substituted amino refers to the group —NRR where each R is independently selected from the group consisting of hydrogen, alkyl, cycloalkyl, carboxyalkyl (for example, benzyloxycarbonyl), aryl, heteroaryl and heterocyclyl provided that both R groups are not hydrogen, or a group —Y-Z, in which Y is optionally substituted alkylene and Z is alkenyl, cycloalkenyl, or alkynyl, Unless otherwise constrained by the definition, all substituents may optionally be further substituted by 1-3 substituents chosen from alkyl, carboxy, carboxyalkyl, aminocarbonyl, hydroxy, alkoxy, halogen, CF 3 , amino, substituted amino, cyano, and —S(O) n R, where R is alkyl, aryl, or heteroaryl and n is 0, 1 or 2.
  • Carboxyalkyl refers to the groups —C(O)O-alkyl or —C(O)O-cycloalkyl, where alkyl and cycloalkyl, are as defined herein, and may be optionally further substituted by alkyl, alkenyl, alkynyl, alkoxy, halogen, CF 3 , amino, substituted amino, cyano, or —S(O) n R, in which R is alkyl, aryl, or heteroaryl and n is 0, 1 or 2.
  • cycloalkyl refers to carbocyclic groups of from 3 to 20 carbon atoms having a single cyclic ring or multiple condensed rings.
  • Such cycloalkyl groups include, by way of example, single ring structures such as cyclopropyl, cyclobutyl, cyclopentyl, cyclooctyl, and the like, or multiple ring structures such as adamantanyl, bicyclo[2.2.1]heptane, 1,3,3-trimethylbicyclo[2.2.1]hept-2-yl, (2,3,3-trimethylbicyclo[2.2.1]hept-2-yl), or carbocyclic groups to which is fused an aryl group, for example indane, and the like.
  • substituted cycloalkyl refers to cycloalkyl groups having 1, 2, 3, 4 or 5 substituents, and preferably 1, 2, or 3 substituents, selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkenyl, acyl, acylamino, acyloxy, amino, aminocarbonyl, alkoxycarbonylamino, azido, cyano, halogen, hydroxy, keto, thiocarbonyl, carboxy, carboxyalkyl, arylthio, heteroarylthio, heterocyclylthio, thiol, alkylthio, aryl, aryloxy, heteroaryl, aminosulfonyl, aminocarbonylamino, heteroaryloxy, heterocyclyl, heterocyclooxy, hydroxyamino, alkoxyamino, nitro, —SO-alkyl, —SO-aryl
  • substituents may optionally be further substituted by 1, 2, or 3 substituents chosen from alkyl, carboxy, carboxyalkyl, aminocarbonyl, hydroxy, alkoxy, halogen, CF 3 , amino, substituted amino, cyano, and —S(O) n R, where R is alkyl, aryl, or heteroaryl and n is 0, 1 or 2.
  • halogen refers to fluoro, bromo, chloro, and iodo.
  • acyl denotes a group —C(O)R, in which R is hydrogen, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl.
  • heteroaryl refers to a radical derived from an aromatic cyclic group (i.e., fully unsaturated) having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 carbon atoms and 1, 2, 3 or 4 heteroatoms selected from oxygen, nitrogen and sulfur within at least one ring.
  • Such heteroaryl groups can have a single ring (e.g., pyridyl or furyl) or multiple condensed rings (e.g., indolizinyl, benzothiazolyl, or benzothienyl).
  • heteroaryls include, but are not limited to, [1,2,4]oxadiazole, [1,3,4]oxadiazole, [1,2,4]thiadiazole, [1,3,4]thiadiazole, pyrrole, imidazole, pyrazole, pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole, indole, indazole, purine, quinolizine, isoquinoline, quinoline, phthalazine, naphthylpyridine, quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline, phenanthridine, acridine, phenanthroline, isothiazole, phenazine, isoxazole, phenoxazine, phenothiazine, imidazolidine, imidazoline, and the like as well as N-oxide and N-oxid
  • heteroarylene refers to a diradical of a heteroaryl group as defined above. This term is exemplified by groups such as 2,5-imidazolene, 3,5-[1,2,4]oxadiazolene, 2,4-oxazolene, 1,4-pyrazolene, and the like.
  • 1,4-pyrazolene is: where A represents the point of attachment.
  • heteroaryl or heteroarylene groups can be optionally substituted with 1 to 5 substituents, preferably 1 to 3 substituents selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkenyl, acyl, acylamino, acyloxy, amino, aminocarbonyl, alkoxycarbonylamino, azido, cyano, halogen, hydroxy, keto, thiocarbonyl, carboxy, carboxyalkyl, arylthio, heteroarylthio, heterocyclylthio, thiol, alkylthio, aryl, aryloxy, heteroaryl, aminosulfonyl, aminocarbonylamino, heteroaryloxy, heterocyclyl, heterocyclooxy, hydroxyamino, alkoxyamino, nitro, —SO-alkyl,
  • substituents may optionally be further substituted by 1-3 substituents chosen from alkyl, carboxy, carboxyalkyl, aminocarbonyl, hydroxy, alkoxy, halogen, CF 3 , amino, substituted amino, cyano, and —S(O) n R, where R is alkyl, aryl, or heteroaryl and n is 0, 1 or 2.
  • heteroarylkyl refers to a heteroaryl group covalently linked to an alkylene group, where heteroaryl and alkylene are defined herein.
  • Optionally substituted heteroaralkyl refers to an optionally substituted heteroaryl group covalently linked to an optionally substituted alkylene group.
  • Such heteroaralkyl groups are exemplified by 3-pyridylmethyl, quinolin-8-ylethyl, 4-methoxythiazol-2-ylpropyl, and the like.
  • heteroaryloxy refers to the group heteroaryl-O—.
  • heterocyclyl refers to a monoradical saturated or partially unsaturated group having a single ring or multiple condensed rings, having from 1 to 40 carbon atoms and from 1 to 10 hetero atoms, preferably 1, 2, 3 or 4 heteroatoms, selected from nitrogen, sulfur, phosphorus, and/or oxygen within the ring.
  • Heterocyclic groups can have a single ring or multiple condensed rings, and include tetrahydrofuranyl, morpholino, piperidinyl, piperazino, dihydropyridino, and the like.
  • heterocyclic groups can be optionally substituted with 1, 2, 3, 4 or 5, and preferably 1, 2 or 3 substituents, selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkenyl, acyl, acylamino, acyloxy, amino, aminocarbonyl, alkoxycarbonylamino, azido, cyano, halogen, hydroxy, keto, thiocarbonyl, carboxy, carboxyalkyl, arylthio, heteroarylthio, heterocyclylthio, thiol, alkylthio, aryl, aryloxy, heteroaryl, aminosulfonyl, aminocarbonylamino, heteroaryloxy, heterocyclyl, heterocyclooxy, hydroxyamino, alkoxyamino, nitro, —SO-alkyl, —SO-aryl,
  • substituents may optionally be further substituted by 1-3 substituents chosen from alkyl, carboxy, carboxyalkyl, aminocarbonyl, hydroxy, alkoxy, halogen, CF 3 , amino, substituted amino, cyano, and —S(O) n R, where R is alkyl, aryl, or heteroaryl and n is 0, 1 or 2.
  • thiol refers to the group —SH.
  • substituted alkylthio refers to the group —S-substituted alkyl.
  • heteroarylthiol refers to the group —S-heteroaryl wherein the heteroaryl group is as defined above including optionally substituted heteroaryl groups as also defined above.
  • sulfoxide refers to a group —S(O)R, in which R is alkyl, aryl, or heteroaryl. “Substituted sulfoxide” refers to a group —S(O)R, in which R is substituted alkyl, substituted aryl, or substituted heteroaryl, as defined herein.
  • sulfone refers to a group —S(O) 2 R, in which R is alkyl, aryl, or heteroaryl. “Substituted sulfone” refers to a group —S(O) 2 R, in which R is substituted alkyl, substituted aryl, or substituted heteroaryl, as defined herein.
  • keto refers to a group —C(O)—.
  • thiocarbonyl refers to a group —C(S)—.
  • compound of Formula I and Formula II is intended to encompass the compounds of the invention as disclosed, and the pharmaceutically acceptable salts, pharmaceutically acceptable esters, prodrugs, hydrates and polymorphs of such compounds. Additionally, the compounds of the invention may possess one or more asymmetric centers, and can be produced as a racemic mixture or as individual enantiomers or diastereoisomers. The number of stereoisomers present in any given compound of Formula I depends upon the number of asymmetric centers present (there are 2 n stereoisomers possible where n is the number of asymmetric centers).
  • the individual stereoisomers may be obtained by resolving a racemic or non-racemic mixture of an intermediate at some appropriate stage of the synthesis, or by resolution of the compound of Formula I by conventional means.
  • the individual stereoisomers (including individual enantiomers and diastereoisomers) as well as racemic and non-racemic mixtures of stereoisomers are encompassed within the scope of the present invention, all of which are intended to be depicted by the structures of this specification unless otherwise specifically indicated.
  • Steps are isomers that differ only in the way the atoms are arranged in space.
  • Enantiomers are a pair of stereoisomers that are non-superimposable mirror images of each other. A 1:1 mixture of a pair of enantiomers is a “racemic” mixture. The term “( ⁇ )” is used to designate a racemic mixture where appropriate.
  • “Diastereoisomers” are stereoisomers that have at least two asymmetric atoms, but which are not mirror-images of each other.
  • the absolute stereochemistry is specified according to the Cahn-Ingold-Prelog R—S system.
  • the stereochemistry at each chiral carbon may be specified by either R or S.
  • Resolved compounds whose absolute configuration is unknown are designated (+) or ( ⁇ ) depending on the direction (dextro- or laevorotary) which they rotate the plane of polarized light at the wavelength of the sodium D line.
  • Topical administration shall be defined as the delivery of the therapeutic agent to the surface of the wound and adjacent epithelium.
  • Parental administration is the systemic delivery of the therapeutic agent via injection to the patient.
  • therapeutically effective amount refers to that amount of a compound of Formula I that is sufficient to effect treatment, as defined below, when administered to a mammal in need of such treatment.
  • the therapeutically effective amount will vary depending upon the specific activity of the therapeutic agent being used, and the age, physical condition, existence of other disease states, and nutritional status of the patient. Additionally, other medication the patient may be receiving will effect the determination of the therapeutically effective amount of the therapeutic agent to administer.
  • treatment means any treatment of a disease in a mammal, including:
  • the language “genetically and/or environmentally predisposed to airway remodeling and/or pulmonary inflammation” refers to mammals that are susceptible to Examples of such environmental conditions include, but are not limited to, exposure to cigarette smoke and other pollutants, exposure to sprays or chemical agents at work, home, or with hobbies, exposure to common allergens such as dust, grasses, molds, weeds, trees, and animal dander, and exposure to irritants such as asbestos, silica and metal dusts.
  • Examples of genetic predisposition can be evidenced by family history or genetic analysis for suspected mutations in the ADAM33 gene, TLR4 polymorphisms, IL-3 polymorphisms, CD14 C-159T polymorphisms, and the like.
  • Bioinformatic methods of screening for a genetic predisposition have been presented by Tomita et al. (2004) BMC Bioinformatics. 5(1):120.
  • Other conditions and disease states that are known to cause airway remodeling and/or pulmonary inflammation include, but are not limited to, lupus, scleroderma, tuberculosis, and rheumatoid arthritis.
  • the compounds of this invention are capable of forming acid and/or base salts by virtue of the presence of amino and/or carboxyl groups or groups similar thereto.
  • pharmaceutically acceptable salt refers to salts that retain the biological effectiveness and properties of the compounds of Formula I, and which are not biologically or otherwise undesirable.
  • Pharmaceutically acceptable base addition salts can be prepared from inorganic and organic bases. Salts derived from inorganic bases, include by way of example only, sodium, potassium, lithium, ammonium, calcium and magnesium salts.
  • Salts derived from organic bases include, but are not limited to, salts of primary, secondary and tertiary amines, such as alkyl amines, dialkyl amines, trialkyl amines, substituted alkyl amines, di(substituted alkyl) amines, tri(substituted alkyl) amines, alkenyl amines, dialkenyl amines, trialkenyl amines, substituted alkenyl amines, di(substituted alkenyl) amines, tri(substituted alkenyl) amines, cycloalkyl amines, di(cycloalkyl) amines, tri(cycloalkyl) amines, substituted cycloalkyl amines, disubstituted cycloalkyl amine, trisubstituted cycloalkyl amines, cycloalkenyl amines, di(cycloalkeny
  • Suitable amines include, by way of example only, isopropylamine, trimethyl amine, diethyl amine, tri(iso-propyl) amine, tri(n-propyl) amine, ethanolamine, 2-dimethylaminoethanol, tromethamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine, glucosamine, N-alkylglucamines, theobromine, purines, piperazine, piperidine, morpholine, N-ethylpiperidine, and the like.
  • Salts derived from inorganic acids include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like.
  • Salts derived from organic acids include acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, malic acid, malonic acid, succinic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluene-sulfonic acid, salicylic acid, and the like.
  • “pharmaceutically acceptable carrier” includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents and the like. The use of such media and agents for pharmaceutically active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active ingredient, its use in the therapeutic compositions is contemplated. Supplementary active ingredients can also be incorporated into the compositions.
  • the present invention relates to methods of preventing airway remodeling and/or pulmonary inflammation by administration of a therapeutically effective amount of a A 2B adenosine receptor antagonist to a mammal having a condition that produces or is caused by such airway remodeling.
  • a A 2B adenosine receptor antagonist As airway remodeling and pulmonary inflammation are significant components of asthma, pulmonary fibrosis, and COPD, the method of the invention will generally involve administration of an A 2B adenosine receptor antagonist to a patient suffering from either asthma, pulmonary fibrosis, and/or COPD.
  • the A 2B adenosine receptor antagonist is administered systemically as an oral formulation but may also be administered directly to the pulmonary tissue via an inhaler. This administration can be as a single dose or as repeated doses given at multiple designated intervals. It will readily be appreciated by those skilled in the art that the preferred dosage regimen will vary with the patient and severity of the condition being treated.
  • compositions that contain, as the active ingredient, one or more of the compounds of Formula I, or a pharmaceutically acceptable salt or ester thereof, and one or more pharmaceutically acceptable excipients, carriers, including inert solid diluents and fillers, diluents, including sterile aqueous solution and various organic solvents, solubilizers and adjuvants.
  • the compounds of Formula I may be administered alone or in combination with other therapeutic agents.
  • Such compositions are prepared in a manner well known in the pharmaceutical art (see, e.g., Remington's Pharmaceutical Sciences, Mace Publishing Co., Philadelphia, Pa. 17 th Ed. (1985) and “Modern Pharmaceutics”, Marcel Dekker, Inc. 3 rd Ed. (G. S. Banker & C. T. Rhodes, Eds.).
  • Any A 2B adenosine receptor antagonist may be used in the method of the invention.
  • Numerous compounds that antagonize the A 2B receptor are known in the art, as are methods for determining if a specific compound has such activity.
  • a review article by Feoktistov and Baggioni, reports the binding affinity of eight adenosine receptor agonists and eight antagonists for all four subtypes of adenosine receptors. References cited therein provide detailed descriptions of the procedures used. (Robeva et al, (1996) J Drug Dev. Res 39:243-252; Jacobson et al (1996) Drug Dev. Res.
  • Effective methods for determining the binding affinity of a compound for a receptor use a radiolabelled agonist or antagonist and correlation of the binding of that compound to a membrane fraction known to contain that receptor; for example, to determine whether a compound is an A 2B antagonist, the membrane fraction would contain the A 2B adenosine receptor.
  • Another particularly effective procedure for determining whether a compound is an A 2B antagonist is reported in U.S. Pat. No. 5,854,081.
  • a 2B receptor subtype Compounds selective for the A 2B receptor subtype are therefore preferred for the present methods.
  • An example, but not a limitation, of such a compound is 3-n-propylxanthine (enprofylline).
  • Suitable compounds are also disclosed in U.S. Pat. No. 6,545,002.
  • Compounds that antagonize other receptors in addition to the A 2B receptor are also suitable for use in the present invention.
  • One example of such a compound is 1,3-dipropyl-8-(p-acrylic)phenylxanthine.
  • a 2B adenosine receptor antagonists are those disclosed in copending and commonly assigned U.S. Pat. No. 6,825,349 and in copending and commonly assigned U.S. patent application Ser. No. 10/719,102, which published as U.S. Patent Application Publication No. 20040176399.
  • the compounds disclosed in that application have the structure of Formula I and Formula II as presented in the Summary of the Invention above and can be synthesized as described in the references or as detailed below.
  • solvent inert organic solvent or “inert solvent” mean a solvent inert under the conditions of the reaction being described in conjunction therewith [including, for example, benzene, toluene, acetonitrile, tetrahydrofuran (“THF”), dimethylformamide (“DMF”), chloroform, methylene chloride (or dichloromethane), diethyl ether, methanol, pyridine and the like].
  • THF tetrahydrofuran
  • DMF dimethylformamide
  • chloroform chloroform
  • methylene chloride or dichloromethane
  • q.s. means adding a quantity sufficient to achieve a stated function, e.g., to bring a solution to the desired volume (i.e., 100%).
  • the compound of formula (2) is made from the compound of formula (1) by a reduction step.
  • Conventional reducing techniques may be used, for example using sodium dithionite in aqueous ammonia solution; preferably, reduction is carried out with hydrogen and a metal catalyst.
  • the reaction is carried out at in an inert solvent, for example methanol, in the presence of a catalyst, for example 10% palladium on carbon catalyst, under an atmosphere of hydrogen, preferably under pressure, for example at about 30 psi, for about 2 hours.
  • a catalyst for example 10% palladium on carbon catalyst
  • the compound of formula (2) is then reacted with a carboxylic acid of the formula Z-Y—X—CO 2 H in the presence of a carbodiimide, for example 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride.
  • a carbodiimide for example 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride.
  • the reaction is conducted in a protic solvent, for example methanol, ethanol, propanol, and the like, preferably methanol, at a temperature of about 20-30° C., preferably about room temperature, for about 12-48 hours, preferably about 16 hours.
  • a protic solvent for example methanol, ethanol, propanol, and the like, preferably methanol
  • the carboxylic acid of the formula Z-Y—X—CO 2 H is first converted to an acid halide of the formula Z-Y—X—C(O)L, where L is chloro or bromo, by reacting with a halogenating agent, for example thionyl chloride or thionyl bromide, preferably thiony chloride.
  • a halogenating agent for example thionyl chloride or thionyl bromide, preferably thiony chloride.
  • oxalyl chloride, phosphorus pentachloride or phosphorus oxychloride may be used.
  • the reaction is preferably conducted in the absence of a solvent, using excess halogenating agent, for example at a temperature of about 60-80° C., preferably about 70° C., for about 1-8 hours, preferably about 4 hours.
  • the product of formula Z-Y—X—C(O)L is isolated conventionally, for example by removal of the excess halogenating agent under
  • the product is then reacted with a compound of formula (2) in an inert solvent, for example acetonitrile, in the presence of a tertiary base, for example triethylamine.
  • an inert solvent for example acetonitrile
  • a tertiary base for example triethylamine.
  • the reaction is conducted at an initial temperature of about 0 C, and then allowed to warm to 20-30° C., preferably about room temperature, for about 12-48 hours, preferably about 16 hours.
  • the product of formula (3) is isolated conventionally, for example by diluting the reaction mixture with water, filtering off the product, and washing the product with water followed by ether.
  • the compound of formula (3) is then converted into a compound of Formula I by a cyclization reaction.
  • the reaction is conducted in a protic solvent, for example methanol, ethanol, propanol, and the like, preferably methanol, in the presence of a base, for example potassium hydroxide, sodium hydroxide, sodium methoxide, sodium ethoxide, potassium t-butoxide, preferably aqueous sodium hydroxide, at a temperature of about 50-80° C., preferably about 80° C., for about 1-8 hours, preferably about 3 hours.
  • a protic solvent for example methanol, ethanol, propanol, and the like, preferably methanol
  • a base for example potassium hydroxide, sodium hydroxide, sodium methoxide, sodium ethoxide, potassium t-butoxide, preferably aqueous sodium hydroxide, at a temperature of about 50-80° C., preferably about 80° C., for about 1-8 hours, preferably about
  • the compound of formula (1) may be prepared by various methods. One preferred method is shown in Reaction Scheme II. Step 1—Preparation of Formula (5)
  • the compound of formula (4) is either commercially available or prepared by means well known in the art. It is reacted with ethyl cyanoacetate in a protic solvent, for example ethanol, in the presence of a strong base, for example sodium ethoxide. The reaction is carried out at about reflux temperature, for about 4 to about 24 hours. When the reaction is substantially complete, the compound of formula (5) thus produced is isolated conventionally.
  • a protic solvent for example ethanol
  • the compound of formula (5) is reacted with the dimethylacetal of N,N-dimethylformamide in a polar solvent, for example N,N-dimethylformamide.
  • a polar solvent for example N,N-dimethylformamide.
  • the reaction is carried out at about 40° C., for about 1 hour.
  • the compound of formula (6) thus produced is reacted with a compound of formula R 1 Hal, where Hal is chloro, bromo, or iodo, in the presence of a base, for example potassium carbonate.
  • the reaction is carried out at about 80° C., for about 4-24 hour.
  • the product of formula (7) is isolated conventionally, for example by evaporation of the solvents under reduced pressure, and the residue is used in the next reaction with no further purification.
  • the compound of formula (7) is reacted with aqueous ammonia in a polar solvent, for example suspended in methanol.
  • a polar solvent for example suspended in methanol.
  • the reaction is carried out at about room temperature, for about 1-3 days.
  • the product of formula (8) is isolated conventionally, for example by chromatography over a silica gel column, eluting, for example, with a mixture of dichloromethane/methanol.
  • the compound of formula (8) is then mixed with sodium nitrite in an aqueous acidic solvent, preferably acetic acid and water, for example 50% acetic acid/water.
  • an aqueous acidic solvent preferably acetic acid and water, for example 50% acetic acid/water.
  • the reaction is carried out at a temperature of about 50-90° C., preferably about 70° C., for about 1 hour.
  • the product of formula (1) is isolated by conventional means.
  • reaction may be conducted in an aqueous solvent, for example dimethylformamide and water, and reacted with a strong acid, for example hydrochloric acid.
  • aqueous solvent for example dimethylformamide and water
  • a strong acid for example hydrochloric acid
  • a compound of formula (8) can be prepared from a compound of formula (10) using a similar method, as shown in Reaction Scheme IIA. Step 2 and 3—Preparation of Formula (7)
  • the compound of formula (10) is reacted with the dimethylacetal of N,N-dimethylformamide in a polar solvent, for example N,N-dimethylformamide.
  • a polar solvent for example N,N-dimethylformamide.
  • the reaction is carried out at about 40° C., for about 1 hour.
  • the compound of formula (6a) thus produced is reacted with a compound of formula R 2 Hal, where Hal is chloro, bromo, or iodo, in the presence of a base, for example potassium carbonate.
  • the reaction is carried out at about 80° C., for about 4-24 hour.
  • the product of formula (7) is isolated conventionally, for example by evaporation of the solvents under reduced pressure, and the residue is used in the next reaction with no further purification.
  • the compound of formula (7) is reacted with aqueous ammonia in a polar solvent, for example suspended in methanol.
  • a polar solvent for example suspended in methanol.
  • the reaction is carried out at about room temperature, for about 1-3 days.
  • the product of formula (8) is isolated conventionally, for example by chromatography over a silica gel column, eluting, for example, with a mixture of dichloromethane/methanol.
  • the compound of formula (3) may also be prepared by various methods. One preferred method is shown in Reaction Scheme III. Step 1—Preparation of Formula (10)
  • the commercially available compound 6-aminouracil is first silylated, for example by reaction with excess hexamethyldisilazane as a solvent in the presence of a catalyst, for example ammonium sulfate.
  • the reaction is carried out at about reflux temperature, for about 1-10 hours.
  • the silylated compound thus produced is isolated conventionally, and then reacted with a compound of formula R 1 Hal, where Hal is chloro, bromo, or iodo, preferably in the absence of a solvent.
  • the reaction is carried out at about reflux, for about 4-48 hours, preferably about 12-16 hours.
  • the product of formula (10) is isolated by conventional means.
  • the compound of formula (10) is then dissolved in an aqueous acid, for example aqueous acetic acid, and reacted with sodium nitrite.
  • the reaction is carried out at a temperature of about 20-50° C., preferably about 30° C., over about 30 minutes.
  • the product of formula (11) is isolated by conventional means, for example by filtration.
  • the compound of formula (11) is then reduced to a diamino derivative.
  • the compound of formula (11) is dissolved in aqueous ammonia, and then a reducing agent, for example sodium hydrosulfite, added.
  • a reducing agent for example sodium hydrosulfite
  • the reaction is conducted at a temperature of about 70° C.
  • the product of formula (12) is isolated conventionally, for example by filtration of the cooled reaction mixture.
  • the compound of formula (12) is then reacted with a carboxylic acid of the formula Z-Y—X—CO 2 H in the presence of a carbodiimide, for example 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride.
  • a carbodiimide for example 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride.
  • the reaction is conducted at a temperature of about 20-30° C., for about 12-48 hours.
  • the product of formula (13) is isolated conventionally, for example by filtration of the cooled reaction mixture.
  • the carboxylic acid of the formula Z-Y—X—CO 2 H is converted to an acid halide of the formula Z-Y—X—C(O)L, where L is chloro or bromo, by reacting with a halogenating agent, for example thionyl chloride or thionyl bromide; alternatively, phosphorus pentachloride or phosphorus oxychloride may be used.
  • a halogenating agent for example thionyl chloride or thionyl bromide
  • phosphorus pentachloride or phosphorus oxychloride may be used.
  • the reaction is preferably conducted in the absence of a solvent, using excess halogenating agent, for example at a temperature of about 60-80° C., preferably about 70° C., for about 1-8 hours, preferably about 4 hours.
  • excess halogenating agent for example at a temperature of about 60-80° C., preferably about 70° C., for about 1-8 hours, preferably about
  • the product of the formula Z-Y—X—C(O)L is then reacted with a compound of formula (12) in an inert solvent, for example acetonitrile, in the presence of a tertiary base, for example triethylamine.
  • a compound of formula (12) in an inert solvent, for example acetonitrile, in the presence of a tertiary base, for example triethylamine.
  • the reaction is conducted at an initial temperature of about 0 C, and then allowed to warm to 20-30° C., preferably about room temperature, for about 12-48 hours, preferably about 16 hours.
  • the product of formula (13) is isolated conventionally, for example by diluting the reaction mixture with water, filtering off the product, and washing the product with water followed by ether.
  • the compound of formula (13) is reacted with a compound of formula R 2 Hal, where Hal is chloro, bromo, or iodo, in the presence of a base, for example potassium carbonate.
  • a base for example potassium carbonate.
  • the reaction is carried out at about room temperature, for about 4-24 hour, preferably about 16 hours.
  • the product of formula (3) is isolated conventionally, for example by evaporation of the solvents under reduced pressure, and the residue may be purified conventionally, or may be used in the next reaction with no further purification.
  • the compound of formula (5) is then mixed with sodium nitrite in an aqueous acidic solvent, preferably acetic acid and water, for example 50% acetic acid/water.
  • an aqueous acidic solvent preferably acetic acid and water, for example 50% acetic acid/water.
  • the reaction is carried out at a temperature of about 50-90° C., preferably about 70° C., for about 1 hour.
  • the product of formula (14) is isolated by conventional means.
  • reaction may be conducted in an aqueous solvent, for example dimethylformamide and water, and reacted with a strong acid, for example hydrochloric acid.
  • aqueous solvent for example dimethylformamide and water
  • a strong acid for example hydrochloric acid
  • the compound of formula (14) is then reduced to a diamino derivative.
  • the compound of formula (14) is dissolved in aqueous ammonia, and then a reducing agent, for example sodium hydrosulfite, added.
  • a reducing agent for example sodium hydrosulfite
  • the reaction is conducted at a temperature of about 70° C.
  • the product of formula (15) is isolated conventionally, for example by filtration of the cooled reaction mixture.
  • the compound of formula (15) is then reacted with a carboxylic acid of the formula Z-Y—X—CO 2 H in the presence of a carbodiimide, for example 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride.
  • a carbodiimide for example 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride.
  • the reaction is conducted at a temperature of about 20-30° C., for about 12-48 hours, in an inert solvent, for example methanol.
  • an inert solvent for example methanol
  • the carboxylic acid of the formula Z-Y—X—CO 2 H is converted to an acid halide of the formula Z-Y—X—C(O)L, where L is chloro or bromo, by reacting with a halogenating agent, for example thionyl chloride or thionyl bromide; alternatively, phosphorus pentachloride or phosphorus oxychloride may be used.
  • a halogenating agent for example thionyl chloride or thionyl bromide
  • phosphorus pentachloride or phosphorus oxychloride may be used.
  • the reaction is preferably conducted in the absence of a solvent, using excess halogenating agent, for example at a temperature of about 60-80° C., preferably about 70° C., for about 1-8 hours, preferably about 4 hours.
  • excess halogenating agent for example at a temperature of about 60-80° C., preferably about 70° C., for about 1-8 hours, preferably about
  • the product of the formula Z-Y—X—C(O)L is then reacted with a compound of formula (15) in an inert solvent, for example acetonitrile, in the presence of a tertiary base, for example triethylamine.
  • a compound of formula (15) in an inert solvent, for example acetonitrile, in the presence of a tertiary base, for example triethylamine.
  • the reaction is conducted at an initial temperature of about 0 C, and then allowed to warm to 20-30° C., preferably about room temperature, for about 12-48 hours, preferably about 16 hours.
  • the product of formula (16) is isolated conventionally, for example by diluting the reaction mixture with water, filtering off the product, and washing the product with water followed by ether.
  • the compound of formula (16) is reacted with a compound of formula R 1 Hal, where Hal is chloro, bromo, or iodo, in the presence of a base, for example potassium carbonate.
  • a base for example potassium carbonate.
  • the reaction is carried out at about 80° C., for about 4-24 hour, preferably about 16 hours.
  • the product of formula (3) is isolated conventionally, for example by evaporation of the solvents under reduced pressure, and the residue may be purified conventionally, or may be used in the next reaction with no further purification.
  • the method and pharmaceutical compositions of the invention are effective in the prevention of airway remodeling and/or pulmonary inflammation in a mammal that is predisposed to airway remodeling and/or pulmonary inflammation.
  • the predisposition may be due to genetic abnormalities, disease states, and/or environmental conditions that have been shown to induce airway remodeling and/or pulmonary inflammation.
  • the compounds of Formula I may be administered in either single or multiple doses by any of the accepted modes of administration of agents having similar utilities, for example as described in those patents and patent applications incorporated by reference, including buccal, intranasal, intra-arterial injection, intravenously, intraperitoneally, parenterally, intramuscularly, subcutaneously, orally, or as an inhalant.
  • Oral administration is the preferred route for administration of the compounds of Formula I.
  • Administration may be via capsule or enteric coated tablets, or the like.
  • the active ingredient is usually diluted by an excipient and/or enclosed within such a carrier that can be in the form of a capsule, sachet, paper or other container.
  • the excipient serves as a diluent, in can be a solid, semi-solid, or liquid material (as above), which acts as a vehicle, carrier or medium for the active ingredient.
  • compositions can be in the form of tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as a solid or in a liquid medium), ointments containing, for example, up to 10% by weight of the active compound, soft and hard gelatin capsules, sterile injectable solutions, and sterile packaged powders.
  • excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum acacia, calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, sterile water, syrup, and methyl cellulose.
  • the formulations can additionally include: lubricating agents such as talc, magnesium stearate, and mineral oil; wetting agents; emulsifying and suspending agents; preserving agents such as methyl- and propylhydroxy-benzoates; sweetening agents; and flavoring agents.
  • compositions of the invention can be formulated so as to provide quick, sustained or delayed release of the active ingredient after administration to the patient by employing procedures known in the art.
  • Controlled release drug delivery systems for oral administration include osmotic pump systems and dissolutional systems containing polymer-coated reservoirs or drug-polymer matrix formulations. Examples of controlled release systems are given in U.S. Pat. Nos. 3,845,770; 4,326,525; 4,902514; and 5,616,345.
  • Another formulation for use in the methods of the present invention employs transdermal delivery devices (“patches”). Such transdermal patches may be used to provide continuous or discontinuous infusion of the compounds of the present invention in controlled amounts.
  • the construction and use of transdermal patches for the delivery of pharmaceutical agents is well known in the art. See, e.g., U.S. Pat. Nos. 5,023,252, 4,992,445 and 5,001,139. Such patches may be constructed for continuous, pulsatile, or on demand delivery of pharmaceutical agents.
  • Adenosine A 2B receptor antagonists such as the compounds of Formula I are effective over a wide dosage range and is generally administered in a pharmaceutically effective amount.
  • each dosage unit contains from 1 mg to 2 g of an adenosine A 2B receptor antagonist, more commonly from 1 to 700 mg, and for parenteral administration, from 1 to 700 mg of an adenosine A 2B receptor antagonist, more commonly about 2 to 200 mg.
  • the amount of the adenosine A 2B receptor antagonist actually administered will be determined by a physician, in the light of the relevant circumstances, including the condition to be treated, the chosen route of administration, the actual compound administered and its relative activity, the age, weight, and response of the individual patient, the severity of the patient's symptoms, and the like.
  • the principal active ingredient is mixed with a pharmaceutical excipient to form a solid preformulation composition containing a homogeneous mixture of a compound of the present invention.
  • a pharmaceutical excipient for preparing solid compositions such as tablets, the principal active ingredient is mixed with a pharmaceutical excipient to form a solid preformulation composition containing a homogeneous mixture of a compound of the present invention.
  • these preformulation compositions as homogeneous, it is meant that the active ingredient is dispersed evenly throughout the composition so that the composition may be readily subdivided into equally effective unit dosage forms such as tablets, pills and capsules.
  • the tablets or pills of the present invention may be coated or otherwise compounded to provide a dosage form affording the advantage of prolonged action, or to protect from the acid conditions of the stomach.
  • the tablet or pill can comprise an inner dosage and an outer dosage component, the latter being in the form of an envelope over the former.
  • the two components can be separated by an enteric layer that serves to resist disintegration in the stomach and permit the inner component to pass intact into the duodenum or to be delayed in release.
  • enteric layers or coatings such materials including a number of polymeric acids and mixtures of polymeric acids with such materials as shellac, cetyl alcohol, and cellulose acetate.
  • compositions for inhalation or insufflation include solutions and suspensions in pharmaceutically acceptable, aqueous or organic solvents, or mixtures thereof, and powders.
  • the liquid or solid compositions may contain suitable pharmaceutically acceptable excipients as described supra.
  • the compositions are administered by the oral or nasal respiratory route for local or systemic effect.
  • Compositions in preferably pharmaceutically acceptable solvents may be nebulized by use of inert gases. Nebulized solutions may be inhaled directly from the nebulizing device or the nebulizing device may be attached to a face mask tent, or intermittent positive pressure breathing machine. Solution, suspension, or powder compositions may be administered, preferably orally or nasally, from devices that deliver the formulation in an appropriate manner.
  • a solution of sodium ethoxide was prepared from sodium (4.8 g, 226 mmol) and dry ethanol (150 ml). To this solution was added amino-N-ethylamide (10 g, 113 mmol) and ethyl cyanoacetate (12.8 g, 113 mmol). This reaction mixture was stirred at reflux for 6 hours, cooled, and solvent removed from the reaction mixture under reduced pressure. The residue was dissolved in water (50 ml), and the pH adjusted to 7 with hydrochloric acid.
  • Example 16A Following the procedure of Example 16A, but replacing 6-amino-1-ethyl-5-nitroso-1,3-dihydropyrimidine-2,4-dione with 6-amino-1-methyl-5-nitroso-1,3-dihydropyrimidine-2,4-dione, 5,6-diamino-1-methyl-1,3-dihydropyrimidine-2,4-dione was prepared.
  • Example 18A Following the procedure of Example 18A, but replacing N-(6-amino-1-ethyl-2,4-dioxo(1,3-dihydropyrimidin-5-yl))(1- ⁇ [3-(trifluoromethyl)phenyl]-methyl ⁇ pyrazol-3-yl)carboxamide with N-(6-amino-1-methyl-2,4-dioxo(1,3-dihydropyrimidin-5-yl)), N-(6-amino-1-methyl-2,4-dioxo-3-propyl(1,3-dihydropyrimidin-5-yl))(1- ⁇ [3-(trifluoromethyl)phenyl]methyl ⁇ pyrazol-4-yl)carboxamide was prepared.
  • HEK-A2B cells Human A 2B adenosine receptor cDNA was stably transfected into HEK-293 cells (referred to as HEK-A2B cells). Monolayers of HEK-A2B cells were washed with PBS once and harvested in a buffer containing 10 mM HEPES (pH 7.4), 10 mM EDTA and protease inhibitors. These cells were homogenized in polytron for 1 minute at setting 4 and centrifuged at 29000 g for 15 minutes at 4° C.
  • the cell pellets were washed once with a buffer containing 10 mM HEPES (pH7.4), 1 mM EDTA and protease inhibitors, and were resuspended in the same buffer supplemented with 10% sucrose. Frozen aliquots were kept at ⁇ 80° C.
  • CHO-A 1 , HEK-A 2A , CHO-A 3 Human A 1 , A 2A , A 3 adenosine receptor cDNAs were stably transfected into either CHO or HEK-293 cells (referred to as CHO-A 1 , HEK-A 2A , CHO-A 3 ). Membranes were prepared from these cells using the same protocol as described above.
  • Competition assays were started by mixing 0.5 nM 3 H-CPX (for CHO-A 1 ), 2 nM 3 H-ZM214385 (HEK-A 2A ) or 0.1 nM 125 I-AB-MECA (CHO-A 3 ) with various concentrations of test compounds and the perspective membranes in TE buffer (50 mM Tris and 1 mM EDTA of CHO-A 1 and HEK-A 2 A) or TEM buffer (50 mM Tris, 1 mM EDTA and 10 mM MgCl 2 for CHO-A 3 ) supplemented with 1 Unit/mL adenosine deaminase.
  • TE buffer 50 mM Tris and 1 mM EDTA of CHO-A 1 and HEK-A 2 A
  • TEM buffer 50 mM Tris, 1 mM EDTA and 10 mM MgCl 2 for CHO-A 3
  • the assays were incubated for 90 minutes, stopped by filtration using Packard Harvester and washed four times with ice-cold TM buffer (10 mM Tris, 1 mM MgCl 2 , pH 7.4). Non specific binding was determined in the presence of 1 ⁇ M CPX (CHO-A 1 ), 1 ⁇ M ZM241385 (HEK-A 2A ) and 1 ⁇ M IB-MECA (CHO-A 3 ). The affinities of compounds (i.e. Ki values) were calculated using GraphPadTM software.
  • Monolayer of transfected cells were collected in PBS containing 5 mM EDTA. Cells were washed once with DMEM and resuspended in DMEM containing 1 Unit/mL adenosine deaminase at a density of 100,000-500,000 cells/ml. 100 ⁇ l of the cell suspension was mixed with 25 ⁇ l containing various agonists and/or antagonists and the reaction was kept at 37° C. for 15 minutes. At the end of 15 minutes, 125 ⁇ l 0.2N HCl was added to stop the reaction. Cells were centrifuged for 10 minutes at 1000 rpm. 100 ⁇ l of the supernatant was removed and acetylated.
  • a 2A and A 2B adenosine receptors are coupled to Gs proteins and thus agonists for A 2A adenosine receptor (such as CGS21680) or for A 2B adenosine receptor (such as NECA) increase the cAMP accumulations whereas the antagonists to these receptors prevent the increase in cAMP accumulations-induced by the agonists.
  • a 1 and A 3 adenosine receptors are coupled to Gi proteins and thus agonists for A 1 adenosine receptor (such as CPA) or for A 3 adenosine receptor (such as IB-MECA) inhibit the increase in cAMP accumulations-induced by forskolin. Antagonists to A 1 and A 3 receptors prevent the inhibition in cAMP accumulations.
  • the model system being used is the adenosine deaminase (ADA)-deficient mouse model of adenosine-dependent pulmonary injury.
  • ADA adenosine deaminase
  • elevations in adenosine are associated with increased pulmonary inflammation and airway remodeling.
  • Many of the features seen in these mice resemble those observed in patients with various forms of chronic lung disease including severe asthma, COPD and pulmonary fibrosis.
  • mice with the A 2B AR antagonist 3-ethyl-1-propyl-8-(1- ⁇ [3-(trifluoromethyl)phenyl]methyl ⁇ (4-hydropyrazol-4-yl))-1,3,7-trihydropurine-2,6-dione as a means to probe A 2B AR contributions to pulmonary inflammation and injury in ADA-deficient mice, which should provide insight into the efficacy of this drug for the treatment of chronic lung diseases.
  • a 2B AR antagonist 3-ethyl-1-propyl-8-(1- ⁇ [3-(trifluoromethyl)phenyl]methyl ⁇ (4-hydropyrazol-4-yl))-1,3,7-trihydropurine-2,6-dione as a means to probe A 2B AR contributions to pulmonary inflammation and injury in ADA-deficient mice, which should provide insight into the efficacy of this drug for the treatment of chronic lung diseases.
  • the specific protocol is as follows: ADA-deficient (ADA ⁇ / ⁇ ) or ADA containing (ADA+) mice were identified at birth by screening of ADA enzymatic activity in the blood. ADA ⁇ / ⁇ mice were maintained on ADA enzyme therapy from postnatal day 2 until postnatal day 21.
  • Treatment groups included ADA ⁇ / ⁇ or ADA+ mice receiving 3-ethyl-1-propyl-8-(1- ⁇ [3-(trifluoromethyl)phenyl]methyl ⁇ (4-hydropyrazol-4-yl))-1,3,7-trihydropurine-2,6-dione, vehicle, or no treatment. All mice were littermates and were therefore strain matched. Both males and females were included in these experiments.
  • ADA-deficient mice were generated and genotyped as described (Blackburn et al. (1998) J Biol Chem 273:5093-5100 and Young et al. (2004) J. Immunol. 173:1380-1389. Mice homozygous for the null Ada allele were designated ADA-deficient (ADA ⁇ / ⁇ ), while mice heterozygous for the null Ada allele were designated as ADA control mice (ADA + ). All mice were on a mixed 129sv/C57BL/6J background and all phenotypic comparisons were performed amongst littermates. Animal care was in accordance with institutional and NIH guidelines. Mice were housed in ventilated cages equipped with microisolator lids and maintained under strict containment protocols. No evidence of bacterial, parasitic, or fungal infection was found, and serologies on cage littermates were negative for 12 of the most common murine viruses.
  • mice were anesthetized with avertin, and lungs were lavaged four times with 0.3 ml PBS, and 0.95-1 ml of pooled lavage fluid was recovered. Total cell counts were determined using a hemocytometer, and aliquots were cytospun onto microscope slides and stained with Diff-Quick (Dade Nehring) for cellular differentials. Lungs were then infused with 4% paraformaldehyde in PBS at 25 cm of pressure and then fixed overnight at 4° C. Fixed lung samples were rinsed in PBS, dehydrated, and embedded in paraffin. Sections (5 ⁇ m) were collected on microscope slides and stained with hematoxylin and eosin (H&E; Shandon-Lipshaw) or Masson's trichrome (EM Science), according to manufacturer's instructions.
  • H&E hematoxylin and eosin
  • EM Science Masson's trichrome
  • RNA samples were then DNase treated and subjected to quantitative real-time RT-PCR.
  • the primers, probes and procedures for real-time RT-PCR were described previously in Sun et al. (2005) J Clin Invest 115:3543. Reactions were carried out on a Smart Cycler rapid thermal cycler system (Cepheid, Sunnyvale, Calif.). Specific transcript levels were determined using Smart Cycler analysis software through comparison to a standard curve generated from the PCR amplification of template dilutions.
  • the Sircol collagen assay (Biocolor Ltd., Harbor N. Ireland) was performed on snap frozen whole lungs. Lungs were homogenized in 5 ml. 0.5M Acetic acid with 20 mg of pepsin and incubated with shaking for 24 hrs at 4° C. Homogenate was spun at 4000 rpm and supernatant was assayed for pepsin soluble collagen according to manufacture's instructions.
  • Immunohistochemistry was performed on 5 ⁇ m sections cut from formalin-fixed, paraffin embedded lungs. Sections were rehydrated through graded ethanols to water, endogenous peroxidases were quenched with 3% hydrogen peroxide, antigen retrieval was performed (DAKO Corp., Carpenteria, Calif.), and endogenous avidin and biotin was blocked with the Biotin Blocking System (DAKO Corp.).
  • ⁇ -smooth muscle actin (sma) staining slides were processed with the Mouse on Mouse Kit, and the ABC Elite Streptavidin Reagents (Vector Laboratories, Burlingame, Calif.) and incubated with a 1:500 dilution of a ⁇ -sma monoclonal antibody (Sigma, monoclonal clonel A-4) overnight at 4° C. Sections were developed with DAB (Sigma) and counterstained with Methyl Green.
  • the size of alveolar airways was determined in pressure infused lungs by measuring mean chord lengths on H&E-stained lung sections (Blackburn et al. (2000) J Exp Med 192:159-170). Representative images were digitized, and a grid consisting of 53 black lines at 10.5- ⁇ m intervals was overlaid on the image. This line grid was subtracted from the lung images using Image-Pro® Plus (Media Cybernetics) image analysis software, and the resultant lines were measured and averaged to give the mean chord length of the alveolar airways. The final mean chord lengths represent averages from 10 non-overlapping images of each lung specimen. All quantitative studies were performed blinded with regards to animal genotype.
  • FIG. 1 Histological analysis of lungs are shown in FIG. 1 .
  • the lungs of ADA ⁇ / ⁇ mice treated with vehicle exhibited significant alveolar airway simplification ( FIG. 1B ) and increased pulmonary inflammation that consisted predominantly of accumulation of activated macrophages in the distal airways ( FIG. 1E ).
  • FIG. 1B The lungs of ADA ⁇ / ⁇ mice treated with vehicle exhibited significant alveolar airway simplification
  • FIG. 1E increased pulmonary inflammation that consisted predominantly of accumulation of activated macrophages in the distal airways
  • peribronchial/perivascular inflammation was also evident (not shown).
  • Alveolar airway simplification and pulmonary inflammation was not evident in ADA+ vehicle treated ( FIGS.
  • Bronchiolalveolar lavage (BAL) was performed and airway cell counts and differentials were determined ( FIG. 2 ). Results revealed that there was a significant reduction in the number of total cells recovered from BAL of ADA ⁇ / ⁇ mice treated with the A 2B adenosine receptor antagonist as compared to vehicle treated ADA ⁇ / ⁇ mice ( FIG. 2A ). Analysis of cellular differentials from BALs revealed a reduction in all cell types examined including lymphocytes, neutrophils, eosinophils ( FIG. 2C ) and alveolar macrophages ( FIG. 2B ) in ADA ⁇ / ⁇ mice treated with the A 2B adenosine receptor antagonist.
  • FIGS. 1E and F Examination of alveolar macrophages within the lungs of ADA ⁇ / ⁇ mice with and without A 2B adenosine receptor antagonist treatment indicate that there was a difference in the degree of alveolar macrophage activation (compare FIGS. 1E and F). These observations were confirmed by quantifying the number of activated macrophages recovered from the BAL ( FIG. 2C ). In addition, reduction in alveolar macrophage activation can be appreciated by directly examining BAL cells cytospun onto microscope slides ( FIG. 3 ).
  • adenosine A 2B antagonist treatment to dampen the degree of pulmonary inflammation in ADA ⁇ / ⁇ mice prompted the examination of the levels of key cytokines and chemokines.
  • Whole lung RNA extracts from ADA + and ADA ⁇ / ⁇ mice treated with vehicle or an A 2B adenosine receptor antagonist were analyzed.
  • IL-5, IL-4, TNF ⁇ , RANTES and various monocyte chemoatractant proteins (MCPs) were found to be elevated in the lungs of ADA ⁇ / ⁇ mice treated with vehicle; however their levels did not change with A 2B adenosine receptor antagonist treatment.
  • IL-6, Eotaxin I and TARC were elevated in vehicle treated ADA ⁇ / ⁇ mice and their levels were significantly lower in ADA ⁇ / ⁇ mice treated with the A 2B adenosine receptor antagonist.
  • FIG. 4 these findings demonstrate that A 2B AR antagonism in ADA-1 mice is able to prevent the expression of certain but not all pro-inflammatory cytokines and chemokines.
  • TGF- ⁇ 1, osteopontin (OPN) and plasminogen activator inhibitor-1 (PAI-1) are pro-fibrotic mediators that have been shown to be elevated in the lungs of ADA ⁇ / ⁇ mice (Chunn et al (2005) J Immunol 175:1937-1946). The levels of these fibrotic regulators were elevated in the lungs of ADA ⁇ / ⁇ mice treated with vehicle, while adenosine A 2B receptor antagonist treatment decreased expression of these molecules ( FIG. 7 ). These findings suggest that A 2B AR antagonism can prevent the expression of key regulators of fibrosis in the lungs of ADA ⁇ / ⁇ mice.
  • MMPs metalloproteinases
  • inhibitors of proteases are features of distal airway enlargement in many models including ADA ⁇ / ⁇ mice (Sun et al. (2005) J Clin Invest 115:35-43, Hautamaki et al. (1997) Science 277:2002-2004, Lanone et al. (2002) J Clin Invest 110:463474).
  • Examination of anti-proteases and proteases in the lungs of ADA ⁇ / ⁇ mice treated with vehicle demonstrated an increase in the expression of TIMP-1, MMP-9 and MMP-12 ( FIG. 8 ).
  • ADA ⁇ / ⁇ mice develop features of distal airway enlargement characteristic of emphysema that are mediated by elevations in lung adenosine levels.
  • ADA ⁇ / ⁇ airways were enlarged ( FIG. 9B ) and treatment of ADA ⁇ / ⁇ mice with 3-ethyl-1-propyl-8-(1- ⁇ [3-(trifluoromethyl)phenyl]methyl ⁇ (4-hydropyrazol-4-yl))-1,3,7-trihydropurine-2,6-dione prevented this enlargement ( FIG. 9C ). Quantification of distal airway size agreed with the histological observations ( FIG. 9D ).
US11/251,450 2004-10-15 2005-10-14 Method of preventing and treating airway remodeling and pulmonary inflammation using A2B adenosine receptor antagonists Abandoned US20060159627A1 (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
US11/251,450 US20060159627A1 (en) 2004-10-15 2005-10-14 Method of preventing and treating airway remodeling and pulmonary inflammation using A2B adenosine receptor antagonists
US12/605,783 US8466129B2 (en) 2004-10-15 2009-10-26 Method of preventing and treating airway remodeling and pulmonary inflammation using A2B adenosine receptor antagonists

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US61943904P 2004-10-15 2004-10-15
US11/251,450 US20060159627A1 (en) 2004-10-15 2005-10-14 Method of preventing and treating airway remodeling and pulmonary inflammation using A2B adenosine receptor antagonists

Related Child Applications (1)

Application Number Title Priority Date Filing Date
US12/605,783 Continuation US8466129B2 (en) 2004-10-15 2009-10-26 Method of preventing and treating airway remodeling and pulmonary inflammation using A2B adenosine receptor antagonists

Publications (1)

Publication Number Publication Date
US20060159627A1 true US20060159627A1 (en) 2006-07-20

Family

ID=35677798

Family Applications (2)

Application Number Title Priority Date Filing Date
US11/251,450 Abandoned US20060159627A1 (en) 2004-10-15 2005-10-14 Method of preventing and treating airway remodeling and pulmonary inflammation using A2B adenosine receptor antagonists
US12/605,783 Active 2026-06-08 US8466129B2 (en) 2004-10-15 2009-10-26 Method of preventing and treating airway remodeling and pulmonary inflammation using A2B adenosine receptor antagonists

Family Applications After (1)

Application Number Title Priority Date Filing Date
US12/605,783 Active 2026-06-08 US8466129B2 (en) 2004-10-15 2009-10-26 Method of preventing and treating airway remodeling and pulmonary inflammation using A2B adenosine receptor antagonists

Country Status (13)

Country Link
US (2) US20060159627A1 (ja)
EP (2) EP2311462A1 (ja)
JP (2) JP2008516969A (ja)
KR (1) KR20070063548A (ja)
CN (1) CN101039677A (ja)
AU (1) AU2005295654B2 (ja)
CA (1) CA2583986A1 (ja)
IL (1) IL182493A0 (ja)
MX (1) MX2007004373A (ja)
NO (1) NO20072466L (ja)
NZ (2) NZ589657A (ja)
RU (1) RU2391103C2 (ja)
WO (1) WO2006044610A1 (ja)

Cited By (20)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20070066631A1 (en) * 2005-02-25 2007-03-22 Guoquan Wang Pyrazolyl substituted xanthines
US20070249598A1 (en) * 2005-02-25 2007-10-25 Guoquan Wang Derivatives of 8-substituted xanthines
US20080004292A1 (en) * 2006-06-16 2008-01-03 Guoquan Wang Substituted 8-[6-amino-3-pyridyl]xanthines
US20080045549A1 (en) * 2006-06-29 2008-02-21 Pier Giovanni Baraldi Adenosine a2b receptor antagonists
US20080170990A1 (en) * 2006-09-29 2008-07-17 Cv Therapeutics, Inc. Methods for Myocardial Imaging in Patients Having a History of Pulmonary Disease
US20080200456A1 (en) * 2003-08-25 2008-08-21 Guoquan Wang Substituted 8-Heteroaryl Xanthines
US20080213165A1 (en) * 2006-09-01 2008-09-04 Cv Therapeutics, Inc. Methods and Compositions for Increasing Patent Tolerability During Myocardial Imaging Methods
US20080267861A1 (en) * 2007-01-03 2008-10-30 Cv Therapeutics, Inc. Myocardial Perfusion Imaging
US20090007281A1 (en) * 2006-01-13 2009-01-01 Battelle Memorial Institute Animal Model for Assessing Copd-Related Diseases
US20090081120A1 (en) * 2006-09-01 2009-03-26 Cv Therapeutics, Inc. Methods and Compositions for Increasing Patient Tolerability During Myocardial Imaging Methods
US20090317331A1 (en) * 2000-02-23 2009-12-24 Cv Therapeutics, Inc. Method of Identifying Partial Agonists of the A2A Receptor
US20100004445A1 (en) * 2005-02-25 2010-01-07 Pgxhealth, Llc Methods for the synthesis of unsymmetrical cycloalkyl substituted xanthines
US20100086483A1 (en) * 2008-09-29 2010-04-08 Gilead Palo Alto, Inc. Method of multidetector computed tomagraphy
US20100158797A1 (en) * 2004-10-20 2010-06-24 Gilead Palo Alto, Inc. Use of a2a adenosine receptor agonists
US20100160620A1 (en) * 1999-06-22 2010-06-24 Gilead Palo Alto, Inc. N-pyrazole a2a receptor agonists
US20100179313A1 (en) * 2006-02-03 2010-07-15 Gilead Palo Alto, Inc. Process for preparing an a2a-adenosine receptor agonist and its polymorphs
US20100272645A1 (en) * 2002-07-29 2010-10-28 Gilead Palo Alto, Inc. Myocardial perfusion imaging method
WO2012003220A1 (en) * 2010-06-30 2012-01-05 Gilead Sciences, Inc. Use of a2b adenosine receptor antagonists for treating pulmonary hypertension
USRE47351E1 (en) 1999-06-22 2019-04-16 Gilead Sciences, Inc. 2-(N-pyrazolo)adenosines with application as adenosine A2A receptor agonists
WO2022051589A1 (en) * 2020-09-04 2022-03-10 Teon Therapeutics, Inc. Cocrystals of an adenosine a2b receptor antagonist

Families Citing this family (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7625881B2 (en) * 2005-06-16 2009-12-01 Gilead Palo Alto, Inc. Prodrugs of A2B adenosine receptor antagonists
CN102015712A (zh) * 2008-03-26 2011-04-13 阿德维纳斯治疗私人有限公司 作为腺苷受体拮抗剂的杂环化合物
US8859566B2 (en) * 2009-03-13 2014-10-14 Advinus Therapeutics Private Limited Substituted fused pyrimidine compounds
AR085942A1 (es) * 2011-04-07 2013-11-06 Gilead Sciences Inc Uso de receptor de aadenosina para tratar la insuficiencia cardiaca y la arritmia en pacientes posinfarto de miocardio
CN107249630A (zh) * 2014-11-24 2017-10-13 国家健康与医学研究院 通过il‑20r的拮抗作用治疗慢性阻塞性肺疾病的急性加重
ES2580702B1 (es) 2015-02-25 2017-06-08 Palobiofarma, S.L. Derivados de 2-aminopiridina como antagonistas del receptor A2b de adenosina y ligandos del receptor MT3 de melatonina
CA3093234A1 (en) 2018-03-05 2019-09-12 Teon Therapeutics, Inc. Adenosine receptor antagonists and uses thereof

Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6117878A (en) * 1998-02-24 2000-09-12 University Of Virginia 8-phenyl- or 8-cycloalkyl xanthine antagonists of A2B human adenosine receptors
US6387913B1 (en) * 2000-12-07 2002-05-14 S. Jamal Mustafa Method of treating airway diseases with combined administration of A2B and A3 adenosine receptor antagonists
US20030139428A1 (en) * 2001-11-09 2003-07-24 Rao Kalla A2B adenosine receptor antagonists
US20030162764A1 (en) * 2001-12-20 2003-08-28 Arlindo Castelhano Pyrimidine A2b selective antagonist compounds, their synthesis and use
US20030229106A1 (en) * 2001-11-09 2003-12-11 Rao Kalla A2B adenosine receptor antagonists
US20030235555A1 (en) * 2002-04-05 2003-12-25 David Shealey Asthma-related anti-IL-13 immunoglobulin derived proteins, compositions, methods and uses
US20040176399A1 (en) * 2001-11-09 2004-09-09 Elfatih Elzein A2B adenosine receptor antagonists

Family Cites Families (21)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3845770A (en) 1972-06-05 1974-11-05 Alza Corp Osmatic dispensing device for releasing beneficial agent
US4326525A (en) 1980-10-14 1982-04-27 Alza Corporation Osmotic device that improves delivery properties of agent in situ
US5364620A (en) 1983-12-22 1994-11-15 Elan Corporation, Plc Controlled absorption diltiazem formulation for once daily administration
US5023252A (en) 1985-12-04 1991-06-11 Conrex Pharmaceutical Corporation Transdermal and trans-membrane delivery of drugs
US5001139A (en) 1987-06-12 1991-03-19 American Cyanamid Company Enchancers for the transdermal flux of nivadipine
US4992445A (en) 1987-06-12 1991-02-12 American Cyanamid Co. Transdermal delivery of pharmaceuticals
US4902514A (en) 1988-07-21 1990-02-20 Alza Corporation Dosage form for administering nilvadipine for treating cardiovascular symptoms
US5854081A (en) 1996-06-20 1998-12-29 The University Of Patent Foundation Stable expression of human A2B adenosine receptors, and assays employing the same
US6545002B1 (en) 1999-06-01 2003-04-08 University Of Virginia Patent Foundation Substituted 8-phenylxanthines useful as antagonists of A2B adenosine receptors
US6815446B1 (en) 1999-08-31 2004-11-09 Vanderbilt University Selective antagonists of A2B adenosine receptors
DE60001969T2 (de) * 1999-08-31 2004-02-05 Vanderbilt University, Nashville Selektive antagonisten von a2b adenosinrezeptoren
EP1255550A2 (en) 2000-02-17 2002-11-13 Cv Therapeutics, Inc. Method for identifying and using a 2b adenosine receptor antagonists to mediate mammalian cell proliferation
US20080318983A1 (en) 2001-11-09 2008-12-25 Rao Kalla A2b adenosine receptor antagonists
US7317017B2 (en) 2002-11-08 2008-01-08 Cv Therapeutics, Inc. A2B adenosine receptor antagonists
US7304070B2 (en) 2001-11-09 2007-12-04 Cv Therapeutics, Inc. A2B adenosine receptor antagonists
US6933298B2 (en) * 2001-12-08 2005-08-23 Aventis Pharma Deutschland Gmbh Pyridine-2,4-dicarboxylic acid diamides and pyrimidine-4,6-dicarboxylic acid diamides and the use thereof for selectively inhibiting collagenases
ES2229928B1 (es) 2003-10-02 2006-07-01 Almirall Prodesfarma, S.A. Nuevos derivados de pirimidin-2-amina.
ATE419252T1 (de) 2003-10-31 2009-01-15 Cv Therapeutics Inc Antagonisten des a2b-adenosinrezeptors
DK1789053T3 (da) 2004-09-01 2012-08-06 Gilead Sciences Inc Fremgangsmåde til sårheling under anvendelse af a2b-adenosinreceptor-antagonister
US7625881B2 (en) 2005-06-16 2009-12-01 Gilead Palo Alto, Inc. Prodrugs of A2B adenosine receptor antagonists
CN101405003B (zh) 2006-03-17 2011-05-11 吉利德帕洛阿尔托股份有限公司 利用a2b腺苷受体拮抗剂预防和治疗肝病的方法

Patent Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6117878A (en) * 1998-02-24 2000-09-12 University Of Virginia 8-phenyl- or 8-cycloalkyl xanthine antagonists of A2B human adenosine receptors
US6387913B1 (en) * 2000-12-07 2002-05-14 S. Jamal Mustafa Method of treating airway diseases with combined administration of A2B and A3 adenosine receptor antagonists
US20030139428A1 (en) * 2001-11-09 2003-07-24 Rao Kalla A2B adenosine receptor antagonists
US20030229106A1 (en) * 2001-11-09 2003-12-11 Rao Kalla A2B adenosine receptor antagonists
US20040176399A1 (en) * 2001-11-09 2004-09-09 Elfatih Elzein A2B adenosine receptor antagonists
US6977300B2 (en) * 2001-11-09 2005-12-20 Cv Therapeutics, Inc. A2B adenosine receptor antagonists
US20030162764A1 (en) * 2001-12-20 2003-08-28 Arlindo Castelhano Pyrimidine A2b selective antagonist compounds, their synthesis and use
US6916804B2 (en) * 2001-12-20 2005-07-12 Osi Pharmaceuticals, Inc. Pyrimidine A2b selective antagonist compounds, their synthesis and use
US20030235555A1 (en) * 2002-04-05 2003-12-25 David Shealey Asthma-related anti-IL-13 immunoglobulin derived proteins, compositions, methods and uses

Cited By (42)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
USRE47351E1 (en) 1999-06-22 2019-04-16 Gilead Sciences, Inc. 2-(N-pyrazolo)adenosines with application as adenosine A2A receptor agonists
US9045519B2 (en) 1999-06-22 2015-06-02 Gilead Sciences, Inc. N-pyrazole A2A receptor agonists
US20100160620A1 (en) * 1999-06-22 2010-06-24 Gilead Palo Alto, Inc. N-pyrazole a2a receptor agonists
US9163057B2 (en) 2000-02-23 2015-10-20 Gilead Sciences, Inc. Methods of myocardial perfusion imaging
US20090317331A1 (en) * 2000-02-23 2009-12-24 Cv Therapeutics, Inc. Method of Identifying Partial Agonists of the A2A Receptor
US8071566B2 (en) 2000-02-23 2011-12-06 Gilead Sciences, Inc. Methods of coronary imaging
US20100272645A1 (en) * 2002-07-29 2010-10-28 Gilead Palo Alto, Inc. Myocardial perfusion imaging method
US8183226B2 (en) 2002-07-29 2012-05-22 Gilead Sciences, Inc. Myocardial perfusion imaging method
US20100273780A1 (en) * 2003-08-25 2010-10-28 Pgxhealth, Llc Substituted 8-heteroaryl xanthines
US7732455B2 (en) 2003-08-25 2010-06-08 Pgx Health, Llc Substituted 8-heteroaryl xanthines
US20080200456A1 (en) * 2003-08-25 2008-08-21 Guoquan Wang Substituted 8-Heteroaryl Xanthines
US8106029B2 (en) 2004-10-20 2012-01-31 Gilead Sciences, Inc. Use of A2A adenosine receptor agonists
US20100158797A1 (en) * 2004-10-20 2010-06-24 Gilead Palo Alto, Inc. Use of a2a adenosine receptor agonists
US8153628B2 (en) 2005-02-25 2012-04-10 Forest Laboratories Holdings Limited Pyrazolyl substituted xanthines
US20070066631A1 (en) * 2005-02-25 2007-03-22 Guoquan Wang Pyrazolyl substituted xanthines
US20100120765A1 (en) * 2005-02-25 2010-05-13 Pgxhealth, Llc Pyrazolyl substituted xanthines
US20100004445A1 (en) * 2005-02-25 2010-01-07 Pgxhealth, Llc Methods for the synthesis of unsymmetrical cycloalkyl substituted xanthines
US7618962B2 (en) * 2005-02-25 2009-11-17 Pgx Health, Llc Pyrazolyl substituted xanthines
US7579348B2 (en) * 2005-02-25 2009-08-25 Pgxhealth, Llc Derivatives of 8-substituted xanthines
US20070249598A1 (en) * 2005-02-25 2007-10-25 Guoquan Wang Derivatives of 8-substituted xanthines
US8501941B2 (en) 2005-02-25 2013-08-06 Dogwood Pharmaceuticals, Inc. Methods for the synthesis of unsymmetrical cycloakyl substituted xanthines
US8420813B2 (en) 2005-02-25 2013-04-16 Dogwood Pharmaceuticals, Inc. Methods for the synthesis of unsymmetrical cycloalkyl substituted xanthines
US20090007281A1 (en) * 2006-01-13 2009-01-01 Battelle Memorial Institute Animal Model for Assessing Copd-Related Diseases
US8524883B2 (en) 2006-02-03 2013-09-03 Gilead Sciences, Inc. Monohydrate of (1-{9-[4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazol-4-yl)-N-methylcarboxamide
US8268988B2 (en) 2006-02-03 2012-09-18 Gilead Sciences, Inc. Process for preparing an A2A-adenosine receptor agonist and its polymorphs
US20100179313A1 (en) * 2006-02-03 2010-07-15 Gilead Palo Alto, Inc. Process for preparing an a2a-adenosine receptor agonist and its polymorphs
USRE47301E1 (en) 2006-02-03 2019-03-19 Gilead Sciences, Inc. Process for preparing an A2A-adenosine receptor agonist and its polymorphs
US9085601B2 (en) 2006-02-03 2015-07-21 Gilead Sciences, Inc. Process for preparing an A2A-adenosine receptor agonist and its polymorphs
US8106183B2 (en) 2006-02-03 2012-01-31 Gilead Sciences, Inc. Process for preparing an A2A-adenosine receptor agonist and its polymorphs
US7956179B2 (en) 2006-02-03 2011-06-07 Gilead Sciences, Inc. Process for preparing an A2A-adenosine receptor agonist and its polymorphs
US7884100B2 (en) 2006-06-16 2011-02-08 Pgxhealth, Llc Substituted 8-[6-amino-3-pyridyl]xanthines
US20080004292A1 (en) * 2006-06-16 2008-01-03 Guoquan Wang Substituted 8-[6-amino-3-pyridyl]xanthines
US7767685B2 (en) * 2006-06-29 2010-08-03 King Pharmaceuticals Research And Development, Inc. Adenosine A2B receptor antagonists
US20080045549A1 (en) * 2006-06-29 2008-02-21 Pier Giovanni Baraldi Adenosine a2b receptor antagonists
US20080213165A1 (en) * 2006-09-01 2008-09-04 Cv Therapeutics, Inc. Methods and Compositions for Increasing Patent Tolerability During Myocardial Imaging Methods
US20090081120A1 (en) * 2006-09-01 2009-03-26 Cv Therapeutics, Inc. Methods and Compositions for Increasing Patient Tolerability During Myocardial Imaging Methods
US20080170990A1 (en) * 2006-09-29 2008-07-17 Cv Therapeutics, Inc. Methods for Myocardial Imaging in Patients Having a History of Pulmonary Disease
US20080267861A1 (en) * 2007-01-03 2008-10-30 Cv Therapeutics, Inc. Myocardial Perfusion Imaging
US20100086483A1 (en) * 2008-09-29 2010-04-08 Gilead Palo Alto, Inc. Method of multidetector computed tomagraphy
US20120003329A1 (en) * 2010-06-30 2012-01-05 Gilead Sciences, Inc. Use of A2B Adenosine Receptor Antagonists for Treating Pulmonary Hypertension
WO2012003220A1 (en) * 2010-06-30 2012-01-05 Gilead Sciences, Inc. Use of a2b adenosine receptor antagonists for treating pulmonary hypertension
WO2022051589A1 (en) * 2020-09-04 2022-03-10 Teon Therapeutics, Inc. Cocrystals of an adenosine a2b receptor antagonist

Also Published As

Publication number Publication date
WO2006044610A1 (en) 2006-04-27
KR20070063548A (ko) 2007-06-19
AU2005295654B2 (en) 2011-04-14
MX2007004373A (es) 2007-08-08
IL182493A0 (en) 2007-09-20
EP2311462A1 (en) 2011-04-20
NZ554485A (en) 2010-12-24
EP1799221A1 (en) 2007-06-27
US8466129B2 (en) 2013-06-18
JP2008516969A (ja) 2008-05-22
NO20072466L (no) 2007-05-14
NZ589657A (en) 2012-06-29
US20100105706A1 (en) 2010-04-29
RU2391103C2 (ru) 2010-06-10
RU2007117907A (ru) 2008-11-20
CA2583986A1 (en) 2006-04-27
JP2012056965A (ja) 2012-03-22
CN101039677A (zh) 2007-09-19
AU2005295654A1 (en) 2006-04-27

Similar Documents

Publication Publication Date Title
US8466129B2 (en) Method of preventing and treating airway remodeling and pulmonary inflammation using A2B adenosine receptor antagonists
US8609671B2 (en) Method of inhibiting hepatic fibrosis as a result of liver replacement or repair using A2B adenosine receptor antagonists
US7317017B2 (en) A2B adenosine receptor antagonists
US7521554B2 (en) A2B adenosine receptor antagonists
US20050038045A1 (en) A2B adenosine receptor antagonists
AU2012261721A1 (en) Method of preventing and treating hepatic disease using A2B adenosine receptor antagonists
Belardinelli et al. Method of inhibiting hepatic fibrosis as a result of liver replacement or repair using A 2B adenosine receptor antagonists
Zeng et al. Method of treating hepatic disease using A 2B adenosine receptor antagonists
Belardinelli et al. Method of decreasing hepatotoxic side effects using A 2B adenosine receptor antagonists

Legal Events

Date Code Title Description
AS Assignment

Owner name: CV THERAPEUTICS, INC., CALIFORNIA

Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:ZENG, DEWAN;BLACKBUM, MICHAEL;BELARDINELLI, LUIZ;REEL/FRAME:017231/0985;SIGNING DATES FROM 20060112 TO 20060119

AS Assignment

Owner name: CV THERAPEUTICS, INC., CALIFORNIA

Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:ZENG, DEWAN;BELARDINELLI, LUIZ;BLACKBURN, MICHAEL R.;REEL/FRAME:019578/0301;SIGNING DATES FROM 20070716 TO 20070719

STCB Information on status: application discontinuation

Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION

AS Assignment

Owner name: GILEAD PALO ALTO, INC., CALIFORNIA

Free format text: MERGER;ASSIGNORS:APEX MERGER SUB, INC.;CV THERAPEUTICS, INC.;REEL/FRAME:026025/0255

Effective date: 20090417

AS Assignment

Owner name: GILEAD SCIENCES, INC., CALIFORNIA

Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNOR:GILEAD PALO ALTO, INC.;REEL/FRAME:027077/0319

Effective date: 20111006