Inhibitors of hedgehog signal pathways, compositions and uses related thereto

• US 6,867,216 B1
• Filed: 11/08/2000
• Issued: 03/15/2005
• Est. Priority Date: 04/09/1998
• Status: Expired due to Term

• Current Assignee: Johns Hopkins University
• Original Assignee: Johns Hopkins University
• Inventors: Beachy, Philip A., Porter, Jeffrey A., Cooper, Michael K.
• Primary Examiner(s): Badio, Barbara P.
• Application Number:
  US09/708,974
• Time in Patent Office: 1,588 Days
• Field of Search: 514/278
• US Class Current: 514/278
• CPC Class Codes: A61K 31/00 Medicinal preparations containing organic active ingredients
• CPC Class Codes: A61K 31/4355 the heterocyclic ring system containing a five-membered ring having oxygen as a ring hetero atom
• CPC Class Codes: A61K 31/58 containing heterocyclic rings, e.g. danazol, stanozolol, pancuronium or digitogenin digitoxin A61K31/7048

Abstract

The present invention makes availables assays and reagents inhibiting paracrine and/or autocrine signals produced by a hedgehog protein or aberrant activation of a hedgehog signal transduction pathway, e.g., which involve the use of a steroidal alkaloid or other small molecule.

Claims

1. A method for inhibiting unwanted hair growth or inhibiting spermatogenesis, comprising administering to the patient a composition comprising a purified steroidal alkaloid represented in the general formula (IV), or unsaturated forms thereof, and/or pharmaceutically acceptable salts thereof,

wherein

R₂, R₃, R₄, and R₅ independently for each occurrence, represent one or more substituents selected from hydrogen, halogens, alkyls, alkenyls, alkynyls, aryls, hydroxyl, ═

O, ═

S, alkoxyl, silyloxy, amino, nitro, thiol, amines, imines, amides, phosphoryls, phosphonates, phosphines, carbonyls, carboxyls, carboxamides, anhydrides, silyls, ethers, thioethers, alkylsulfonyls, arylsulfonyls, selenoethers, ketones, aldehydes, esters, or —

(CH₂)_m—

R₈;

R₆ is absent or represents halogens, alkyls, alkenyls, alkynyls, aryls, hydroxyl, ═

O, ═

S, alkoxyl, silyloxy, amino, nitro, thiol, amines, imines, amides, phosphoryls, phosphonates, phosphines, carbonyls, carboxyls, carboxamides, anhydrides, silyls, ethers, thioethers, alkylsulfonyls, arylsulfonyls, selenoethers, ketones, aldehydes, esters, or —

(CH₂)_m—

R₈;

R₈ represents an aryl, a cycloalkyl, a cycloalkenyl, a heterocycle, or a polycycle;

R₉ represents one or more substitutions to the ring A or B, which for each occurrence, independently represent halogens, alkyls, alkenyls, alkynyls, aryls, hydroxyl, ═

O, ═

S, alkoxyl, silyloxy, amino, nitro, thiol, amines, imines, amides, phosphoryls, phosphonates, phosphines, carbonyls, carboxyls, carboxamides, anhydrides, silyls, ethers, thioethers, alkylsulfonyls, arylsulfonyls, selenoethers, ketones, aldehydes, esters, or —

(CH₂)_m—

R₈;

R₂₂ is absent or represents an alkyl, an alkoxyl or —

OH; and

m is an integer in the range 0 to 8 inclusive.

3. The method of claim 1 or 2, wherein the steroidal alkaloid does not substantially interfere with the biological activity of such steroids as aldosterone, androstane, androstene, androstenedione, androsterone, cholecalciferol, cholestane, cholic acid, corticosterone, cortisol, cortisol acetate, cortisone, cortisone acetate, deoxycorticosterone, digitoxigenin, ergocalciferol, ergosterol, estradiol-17-α

, estradiol-17-β

, estriol, estrane, estrone, hydrocortisone, lanosterol, lithocholic acid, mestranol, β

-methasone, prednisone, pregnane, pregnenolone, progesterone, spironolactone, testosterone, or triamcinolone.

4. The method of claim 1 or 2, wherein the steroidal alkaloid does not specifically bind a nuclear hormone receptor.

5. The method of claim 1 or 2, wherein the steroidal alkaloid does not specifically bind estrogen or testosterone receptors.

6. The method of claim 1 or 2, wherein the steroidal alkaloid has no estrogenic activity at therapeutic concentrations.

7. The method of claim 1 or 2, wherein the steroidal alkaloid inhibits hedgehog-mediated signal transduction with an ED₅₀ of 1 mM or less.

8. The method of claim 1 or 2, wherein the steroidal alkaloid inhibits hedgehog-mediated signal transduction with an ED₅₀ of 1 μ

M or less.

9. The method of claim 1 or 2, wherein the steroidal alkaloid inhibits hedgehog-mediated signal transduction with an ED₅₀ of 1 nM or less.

10. The method of claim 1 or 2, wherein the steroidal alkaloid is administered as part of a therapeutic or cosmetic application.

11. The method of claim 1 or 2, wherein the steroidal alkaloid is applied as a topical formulation.

2. A method for inhibiting unwanted hair growth or inhibiting spermatogenesis, comprising administering to the patient a composition comprising a purified steroidal alkaloid represented in the general formula (V) or unsaturated forms thereof, and/or pharmaceutically acceptable salts thereof,

wherein

R₂ and R₄ independently for each occurrence, represent one or more substituents selected from hydrogen, halogens, alkyls, alkenyls, alkynyls, aryls, hydroxyl, ═

O, ═

S, alkoxyl, silyloxy, amino, nitro, thiol, amines, imines, amides, phosphoryls, phosphonates, phosphines, carbonyls, carboxyls, carboxamides, anhydrides, silyls, ethers, thioethers, alkylsulfonyls, arylsulfonyls, selenoethers, ketones, aldehydes, esters, or —

(CH₂)_m—

R₈;

R₃ independently for each occurrence, represents one or more substituents selected from hydrogen, halogens, alkyls, alkenyls, alkynyls, aryls, hydroxyl, alkoxyl, silyloxy, amino, nitro, thiol, amines, imines, amides, phosphoryls, phosphonates, phosphines, carbonyls, carboxyls, carboxamides, anhydrides, silyls, ethers, thioethers, alkylsulfonyls, arylsulfonyls, selenoethers, ketones, aldehydes, esters, or —

(CH₂)_m—

R₉;

R₆ is absent or represents halogens, alkyls, alkenyls, alkynyls, aryls, hydroxyl, ═

O, ═

S, alkoxyl, silyloxy, amino, nitro, thiol, amines, imines, amides, phosphoryls, phosphonates, phosphines, carbonyls, carboxyls, carboxamides, anhydrides, silyls, ethers, thioethers, alkylsulfonyls, arylsulfonyls, selenoethers, ketones, aldehydes, esters, or —

(CH₂)_m—

R₈;

R₈ represents an aryl, a cycloalkyl, a cycloalkenyl, a heterocycle, or a polycycle;

R₉ represents one or more substitutions to the ring A or B, which for each occurrence, independently represent halogens, alkyls, alkenyls, alkynyls, aryls, hydroxyl, ═

O, ═

S, alkoxyl, silyloxy, amino, nitro, thiol, amines, imines, amides, phosphoryls, phosphonates, phosphines, carbonyls, carboxyls, carboxamides, anhydrides, silyls, ethers, thioethers, alkylsulfonyls, arylsulfonyls, selenoethers, ketones, aldehydes, esters, or —

(CH₂)_m—

R₈; and

m is an integer in the range 0 to 8 inclusive.