Inhibitors of hedgehog signaling pathways, compositions and uses related thereto

• US 6,432,970 B2
• Filed: 06/04/1998
• Issued: 08/13/2002
• Est. Priority Date: 04/09/1998
• Status: Expired due to Term

• Current Assignee: Johns Hopkins University
• Original Assignee: Johns Hopkins University
• Inventors: Porter, Jeffrey A., Cooper, Michael K., Beachy, Philip A.
• Primary Examiner(s): Badio, Barbara P.
• Application Number:
  US09/090,622
• Publication Number:
  US 20020006931A1
• Time in Patent Office: 1,531 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 comprising contacting a cell sensitive to the hedgehog protein with a steroidal alkaloid, or other small molecule, in a sufficient amount to reduce the sensitivity of the cell to the hedgehog protein.

Claims

1. A method for inhibiting activation of a hedgehog-patched pathway in a patient diagnosed with a hyperproliferative disorder, comprising administering to the patient a composition comprising a purified hedgehog antagonist in a sufficient amount to reduce the activation of the hedgehog-patched pathway in a cell of the patient, wherein the antagonist is a steroidal alkaloid having a structure represented in the general formula (I), or unsaturated forms thereof and/or nor- or homo-derivatives thereof:

wherein, as valence permits,

R₂, R₃, R₄, and R₅, represent one or more substitutions to the ring to which each is attached, for each occurrence, independently represent 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, R₇, and R′

₇, independently for each occurrence, are absent or represent hydrogens, 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, or

R₆ and R₇, or R₇ and R′

₇, taken together form a ring or polycyclic ring, with the proviso that at least one of R₆, R₇, or R′

₇ is present and includes a primary or secondary amine;

R₈ represents an aryl, a cycloalkyl, a cycloaklenyl, a heterocycle, or a polycycle; and

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

2. The method of claim 1, wherein:

R₂ and R₃, for each occurrence, is an —

OH, alkyl, —

O—

alkyl, —

C(O)—

alkyl, or —

C(O)—

R₈;

R₄, for each occurrence represents H, —

OH, =O, alkyl, —

O—

alkyl, —

C(O)—

alkyl, or —

C(O)—

R₈;

R₆, R₇, and R′

₇ each independently represent , hydrogen, alkyls, alkenyls, alkynyls, amines, imines, amides, carbonyls, carboxyls, carboxamides, ethers, thioethers, esters, or —

(CH₂)_m—

R₈, or

R₇, and R′

₇ taken together form a furanopiperidine, such as perhydrofuro[3,2-b]pyridine, a pyranopiperidine, a quinoline, an indole, a pyranopyrrole, a naphthyriding, a thiofuranopiperidine, or a thiopyranopiperidine

with the proviso that at least one of R₆, R₇, or R′

₇ is present and includes a primary or secondary amine;

R₈ represents an aryl, a cycloalkyl, a cycloalkenyl, a heterocycle, or a polycycle, and preferably R₈ is a piperidine, pyrimidine, morpholine, thiomorpholine, pyridazine.

7. The method of claims 1, 3, 4, 5, or 6, wherein the hedgehog antagonist does not substantially interfere with the biological activity of aldosterone, androstane, androstene, androstenedione, androsterone, cholecalciferol, cholestane, cholic acid, corticosterone, cortisol, cortisol acetate, cortisone, cortisone acetate, deoxycorticosterone, digitoxigenen, ergocalciferol, ergosterol, estradiol-17α

, estradiol-17-β

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

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

8. The method of claim 1, 3, 4, 5, or 6, wherein the hedgehog antagonist does not specifically bind a nuclear hormone receptor.

9. The method of claim 1, 3, 4, 5, or 6, wherein the hedgehog antagonist does not specifically bind estrogen or testosterone receptors.

10. The method of claim 1, 3, 4, 5, or 6, wherein the hedgehog antagonist has no estrogenic activity at therapeutic concentrations.

11. The method of claim 1, 3, 4, 5, or 6, wherein the hedgehog antagonist inhibits activation of the hedgehog-patched pathway with an ED₅₀ of 1 mM or less.

12. The method of claim 1, 3, 4, 5, or 6, wherein the hedgehog antagonist inhibits activation of the hedgehog-patched pathway with an ED₅₀ of 1μ

M or less.

13. The method of claim 1, 3, 4, 5, or 6, wherein the hedgehog antagonist inhibits activation of the hedgehog-patched pathway with an ED₅₀ of 1 nM or less.

14. The method of claim 1, 3, 4, 5, ro 6, wherein the hedgehog antagonist is administered as part of a therapeutic or cosmetic application.

15. The method of claim 1, 3, 4, 5, or 6, wherein the hyperproliferative disorder comprises basal cell carcinoma.

16. The method of claim 1, 3, 4, 5, or 6, wherein the hyperproliferative disorder comprises medulloblastoma.

17. The method of claim 1, 3, 4, 5, or 6, wherein the composition is administered topically.

18. The method of claim 1, 3, 4, 5, or 6, wherein the antagonist is other than jervine.

3. A method for inhibiting activation of a hedgehog-patched pathway in a patient diagnosed with a hyperproliferative disorder, comprising administering to the patient a composition comprising a purified hedgehog antagonist in a sufficient amount to reduce the activation of the hedgehog-patched pathway in a cell of the patient, wherein the antagonist is a steroidal alkaloid having a structure represented in the general formula (II), or unsaturated forms thereof and/of nor- or homo-derivatives thereof:

wherein, as valence permits,

R₂, R₃, R₄, and R₅, represent one or more substitutions to the ring to which each is attached, for each occurrence, independently represent 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, R₇, and R′

₇, are absent or represent, independently, halogens, alkyls, alkenyls, alkynyls, aryls, hydroxyl, =O, =S, alkoxyl, silyloxy, amino, nitro, thiol, amines, imines, amides, phosphoryls, phosphonates, phosphines, carbonyls, carboxyls, carboxamides, anhydrides, silyls, etheres, thioethers, alkylsulfonyls, selenoethers, ketones, aldehydes, esters, or —

(CH₂)_m—

R₈, or

R₆ and R₇, or R₇ and R′

₇, taken togehter form a ring or polycyclic ring,

with the proviso that at least one of R₆, R₇, or R′

₇ is present and includes a primary or secondary amine;

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

m is an integer in the range 0 to 8 inclusive; and

X represents O or S.

4. A method for inhibiting activation of a hedgehog-patched pathway in a patient diagnosed with a hyperproliferative disorder, comprising administering to the patient a composition comprising a purified hedgehog antagonist in a sufficient amount to reduce the activation of the hedgehog-patched pathway in a cell of the patient, wherein the antagonist has a structure represented in the general formula (III), or unsaturated forms thereof and/or nor- or homo-derivatives thereof;

wherein, as valence permits,

R₂, R₃, R₄, and R₅, represent one or more substitutions to the ring to which each is attached, for each occurrence, independently represent 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;

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

A and B represent monocyclic or polycyclic groups;

T represents an alkyl, an aminoalkyl, a carboxyl, an ester, an amide, ether or amine linkage of 1-10 bond lengths;

T′

is absent, or represents an alkyl, an aminolkyl, a carboxyl, an ester, an amide, ether of amine linkage of 1-3 bond lengths, wheren if T and T′

are present together, than T and T′

taken together with the ring B form a covalently closed ring of 5-8 ring atoms;

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

n and m are, independently, zero, 1 or 2;

with the proviso that A and R₉, or T, T′

B and R₉, taken together include at least one primary or secondary amine.

5. A method for inhibiting activation of a hedgehog-patched pathway in a patient diagnosed with a hyperproliferative disorder, comprising administering to the patient a composition comprising a purified hedgehog antagonist in a sufficient amount to reduce the activation of the hedgehog-patched pathway in a cell of the patient, wherein the antagonist has a structure represented in the general formula (IV), or unsaturated forms thereof and/or nor- or homo-derivatives thereof:

wherein, as valence permits,

R₂, R₃, R₄, And R₅, represent one or more substitutions to the ring to which each is attached, for each occurrence, independently represent 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, arysulfonyls, 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;

m is 0, 1, or 2; and

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

OH.

6. A method for inhibiting activation of a hedgehog-patched pathway in a patient diagnosed with a hyperproliferative disorder, comprising administering to the patient a composition comprising a purified hedgehog antagonist in a sufficient amount to reduce the activation of the hedgehog-patched pathway in a cell of the patient, wherein the antagonist has a structure represented in the general formula (V) or unsaturated forms thereof and/or nor- or homo-derivatives thereof:

wherein, as valence permits,

R₂, R₃, R₄, and R₅, represent one or more substitutions to the ring to which each is attached, for each occurrence, independently represent 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, selesnoethers, 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, arysulfonyls, selenoethers, ketones, aldehydes, esters, or —

(CH₂)_m—

R₈; and

m is 0, 1, or 2.