Methods for treatment of conditions associated with lactosylceramide

• US 6,228,889 B1
• Filed: 06/25/1999
• Issued: 05/08/2001
• Est. Priority Date: 05/21/1997
• Status: Expired due to Term

• Current Assignee: Johns Hopkins University
• Original Assignee: Johns Hopkins University
• Inventors: Chatterjee, Subroto
• Primary Examiner(s): Higel, Floyd D.
• Assistant Examiner(s): Sackey, Ebenezer
• Application Number:
  US09/340,341
• Time in Patent Office: 683 Days
• Field of Search: 514/625, 514/627, 514/629, 514/237.8, 514/331, 514/428, 424/9.2, 564/224
• US Class Current: 514/629
• CPC Class Codes: A61K 31/16 Amides, e.g. hydroxamic acids
• CPC Class Codes: A61K 31/40 having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
• CPC Class Codes: A61K 31/445 Non condensed piperidines, e.g. piperocaine
• CPC Class Codes: A61K 31/535 having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 9/08 Vasodilators for multiple indications
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis

Abstract

The present invention includes methods for treatment and prophylaxis of diseases, post-surgical disorders and bacterial infections associated with lactosylceramide. The methods generally provide for administration for a mammal, particularly a human, of a therapeutically effective amount of a compound that inhibits UDPGal:GlcCerβ1−>4 galactosylceramide (GalT-2). In vitro and in vivo assays for detecting compounds with therapeutic capacity to modulate GalT-2 are also provided.

Claims

1. A method for determining the therapeutic capacity of a GalT-2 inhibitor compound to reduce restenosis in a mammal, comprising:

performing an invasive surgical procedure on the mammal;

administering a GalT-2 inhibitor compound to the mammal; and

examining a vessel of the mammal for restenosis;

wherein the GalT-2 inhibitor compound is represented by Formula I;

wherein R and R¹ are independently selected from the group consisting of hydrogen and straight-chained or branched C₁-C₆ alkyl with or without a substituent, and further wherein R and R¹ may be taken together to form a 5, 6 or 7-membered ring,

R² is selected from the group consisting of branched or straight-chained C₆-C₃₀ alkyl with or without one to three double bonds, and R³ is selected from the group consisting of straight-chained or branched C₆-C₂₀ alkyl with or without one to three double bonds and aryl or substituted aryl, where the substituent is halo, C₁-C₄ alkoxy, methylenedioxy, C₁-C₄ mercapto, amino or substituted amino in which the amino substituents may suitably be C₁-C₄ alkyl.

2. A method for determining the therapeutic capacity of a GalT-2 inhibitor compound for treating a disease, post-surgical disorder, or bacterial infection modulated by lactosylceramide, the method comprising:

providing a population of cells responsive to lactosylceramide and comprising an oxidase capable of producing an oxygen species;

contacting the cells with lactosylceramide in an amount sufficient to produce the oxygen species;

culturing the cells in medium comprising a GalT-2 inhibitor compound; and

determining effect of the inhibitor compound on amounts of the oxygen species;

wherein the GalT-2 inhibitor compound is represented by Formula I;

wherein R and R¹ are independently selected from the group consisting of hydrogen and straight-chained or branched C₁-C₆ alkyl with or without a substituent, and further wherein R and R¹ may be taken together to form a 5, 6 or 7-membered ring;

R² is selected from the group consisting of branched or straight-chained C₆-C₃₀ alkyl with or without one to three double bonds; and

R³ is selected from the group consisting of straight-chained or branched C₆-C₂₀ alkyl with or without one to three double bonds and aryl or substituted aryl, where the substituent is halo, C_1-C4 alkoxy, methylenedioxy, C₁-C₄ mercapto, amino or substituted amino in which the amino substituents may suitably be C₁-C₄ alkyl.

3. A method for determining therapeutic capacity of a GalT-2 inhibitor compound for treating a disease, post-surgical disorder or bacterial infection modulated by lactosylceramide, the method comprising:

providing a population of cells responsive to lactosylceramide, contacting the cells with lactosylceramide in an amount sufficient to enhance loading of an oncogenic protein to a nucleotide triphosphate;

culturing the cells with a GalT-2 inhibitor compound; and

determining effect of the inhibitor compound on the oncogenic protein loading to the nucleoside triphosphate;

wherein the GalT-2 inhibitor compound is represented by Formula I;

wherein R and R¹ are independently selected from the group consisting of hydrogen and straight-chained or branched C₁-C₆ alkyl with or without a substituent, and further wherein R and R¹ may be taken together to form a 5, 6 or 7-membered ring;

R² is selected from the group consisting of branched or straight-chained C₆-C₃₀ alkyl with or without one to three double bonds; and

R³ is selected from the group consisting of straight-chained or branched C₆-C₂₀ alkyl with or without one to three double bonds and aryl or substituted aryl, where the substituent is halo, C₁-C₄ alkoxy, methylenedioxy, C₁-C₄ mercapto, amino or substituted amino in which the amino substituents may suitably be C₁-C₄ alkyl.

4. A method for determining therapeutic capacity of a GalT-2 inhibitor compound for treating a disease, post-surgical disorder, or bacterial condition modulated by lactosylceramide, the method comprising:

providing a population of cells responsive to lactosylceramide and comprising a protein kinase cascade comprising a mitogen-activated protein kinase;

contacting the cells with lactosylceramide in an amount sufficient to increase phosphorylation of at least one of the protein kinases in the cascade;

culturing the cells in medium comprising the inhibitor compound; and

determining effect of the inhibitor compound on the phosphorylation of the protein kinase;

wherein the GalT-2 inhibitor compound is represented by Formula I;

wherein R and R¹ are independently selected from the group consisting of hydrogen and straight-chained or branched C₁-C₆ alkyl with or without a substituent, and further wherein R and R¹ may be taken together to form a 5, 6 or 7-membered ring;

R² is selected from the group consisting of branched or straight-chained C₆-C₃₀ alkyl with or without one to three double bonds; and

R³ is selected from the group consisting of straight-chained or branched C₆-C₂₀ alkyl with or without one to three double bonds and aryl or substituted aryl, where the substituent is halo, C₁-C₄ alkoxy, methylenedioxy, C₁-C₄ mercapto, amino or substituted amino in which the amino substituents may suitably be C₁-C₄ alkyl.

5. A method for determining therapeutic capacity of a GalT-2 inhibitor compound for treating a disease, post-surgical disorder or bacterial infection modulated by lactosylceramide, the method comprising:

providing a population of cells responsive to lactosylceramide;

contacting the cells with lactosylceramide in an amount sufficient to increase activity of a transcription factor;

culturing the cells in medium comprising the compound; and

determining effect of the inhibitor compound on the transcription factor activity;

wherein the GalT-2 inhibitor compound is represented by Formula I;

wherein R and R¹ are independently selected from the group consisting of hydrogen and straight-chained or branched C₁-C₆ alkyl with or without a substituent, and further wherein R and R¹ may be taken together to form a 5, 6 or 7-membered ring,

R² is selected from the group consisting of branched or straight-chained C₆-C₃₀ alkyl with or without one to three double bonds; and

R³ is selected from the group consisting of straight-chained or branched C₆-C₂₀ alkyl with or without one to three double bonds and aryl or substituted aryl, where the substituent is halo, C₁-C₄ alkoxy, methylenedioxy, C₁-C₄ mercapto, amino or substituted amino in which the amino substituents may suitably be C₁-C₄ alkyl.

6. A method for determining therapeutic capacity of a GalT-2 inhibitor compound for treating a disease, post-surgical disorder, or bacterial infection modulated by lactosylceramide, the method comprising:

providing a first population of cells responsive to lactosylceramide and capable of expressing a receptor which binds an adhesion molecule;

contacting the cells with lactosylceramide in an amount sufficient to increase expression of the receptor in the cells;

culturing the cells in medium comprising the compound;

contacting the cells with a second population of cells expressing the adhesion molecule; and

determining effect of the inhibitor compound on the adhesion

wherein the GalT-2 inhibitor compound is represented by Formula I;

wherein R and R¹ are independently selected from the group consisting of hydrogen and straight-chained or branched C₁-C₆ alkyl with or without a substituent, and further wherein R and R¹ may be taken together to form a 5, 6 or 7-membered ring;

R² is selected from the group consisting of branched or straight-chained C₆-C₃₀ alkyl with or without one to three double bonds; and

R3 is selected from the group consisting of straight-chained or branched C₆-C₂₀ alkyl with or without one to three double bonds and aryl or substituted aryl, where the substituent is halo, C₁-C₄ alkoxy, methylenedioxy, C₁-C₄ mercapto, amino or substituted amino in which the amino substituents may suitably be C₁-C₄ alkyl.

7. A method for determining therapeutic capacity of a GalT-2 inhibitor compound for treating a bacterial infection modulated by binding of a bacterial toxin to lactosylceramide, the method comprising:

providing a population of cells responsive to lactosylceramide;

contacting the cells with a bacterial toxin capable of binding the lactosylceramide;

culturing the cells in medium comprising the compound; and

determining any effect of the inhibitor compound on the cell viability wherein the GalT-2 inhibitor compound is represented by Formula I;

wherein R and R¹ are independently selected from the group consisting of hydrogen and straight-chained or branched C₁-C₆ alkyl with or without a substituent, and further wherein R and R¹ may be taken together to form a 5, 6 or 7-membered ring;

R² is selected from the group consisting of branched or straight-chained C₆-C₃₀ alkyl with or without one to three double bonds; and

R³ is selected from the group consisting of straight-chained or branched C₆-C₂₀ alkyl with or without one to three double bonds and aryl or substituted aryl, where the substituent is halo, C₁-C₄ alkoxy, methylenedioxy, C₁-C₄ mercapto, amino or substituted amino in which the amino substituents may suitably be C₁-C₄ alkyl.

8. A method for determining the therapeutic capacity of a GalT-2 inhibitor compound for treating a disease, post-surgical disorder, or bacterial infection modulated by lactosylceramide, the method comprising:

providing a population of cells responsive to lactosylceramide and comprising a cell proliferation factor;

contacting the cells with lactosylceramide in an amount sufficient to decrease activity of the cell proliferation factor;

culturing the cells in medium comprising the GalT-2 inhibitor compound; and

determining effect of the inhibitor compound on the levels of the cell proliferation factor;

wherein the GalT-2 inhibitor compound is represented by Formula I;

wherein R and R¹ are independently selected from the group consisting of hydrogen and straight-chained or branched C₁-C₆ alkyl with or without a substituent, and further wherein R and R¹ may be taken together to form a 5, 6 or 7-membered ring;

R² is selected from the group consisting of branched or straight-chained C₆-C₃₀ alkyl with or without one to three double bonds; and

R³ is selected from the group consisting of straight-chained or branched C₆-C₂₀ alkyl with or without one to three double bonds and aryl or substituted aryl, where the substituent is halo, C₁-C₄ alkoxy, methylenedioxy, C₁-C₄ mercanto, amino or substituted amino in which the amino substituents may suitably be C₁-C₄ alkyl.