Inhibitors of hedgehog signaling pathways, compositions and uses related thereto

• US 20080058298A1
• Filed: 08/20/2007
• Published: 03/06/2008
• Est. Priority Date: 04/09/1998
• Status: Active Grant

• Current Assignee: Johns Hopkins University
• Original Assignee: Johns Hopkins University
• Inventors: Cooper, Michael, Porter, Jeffrey, Beachy, Philip
• Granted Patent: US 8,097,632 B2
• Time in Patent Office: Days
• US Class Current: 514/176
• CPC Class Codes: A61K 31/00 Medicinal preparations containing organic active ingredients
• CPC Class Codes: A61K 31/435 having six-membered rings with one nitrogen as the only ring hetero atom
• 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 31/438 The ring being spiro-condensed with carbocyclic or heterocyclic ring systems
• CPC Class Codes: A61K 31/58 containing heterocyclic rings, e.g. danazol, stanozolol, pancuronium or digitogenin digitoxin A61K31/7048
• CPC Class Codes: A61P 15/16 Masculine contraceptives
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 35/00 Antineoplastic agents

Abstract

The present invention makes available 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 paracrine and/or autocrine signals produced by a hedgehog proteins comprising contacting a cell sensitive to the hedgehog protein with a hedgehog antagonist in a sufficient amount to reduce the sensitivity of the cell to the hedgehog protein, wherein the hedgehog antagonist is a organic molecule having a molecule weight less than 750 amu.

3. The method of claim 1 or 2, wherein hedgehog antagonist is a the steroidal alkaloid represented in the general formulas (I), or unsaturated forms thereof and/or seco-, nor- or homo-derivatives thereof:

4. The method of claim 3, wherein:

R₂ and R₃, for each occurrence, is an —

OH, alkyl, —

O-alkyl, —

C(O)-alkyl, or —

C(O)—

R₈;

R₄, for each occurrence, is an absent, or represents —

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 naphthyridine, 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,

5. The method of claim 1 or 2, wherein the hedgehog antagonist is a steroidal alkaloid represented in the general formula (II), or unsaturated forms thereof and/or seco-, nor- or homo-derivatives thereof:

6. The method of claim 1 or 2, wherein the steroidal hedgehog antagonist is a alkaloid represented in the general formula (III), or unsaturated forms thereof and/or seco-, nor- or homo-derivatives thereof:

7. The method of claim 1 or 2, wherein the hedgehog antagonist is a steroidal alkaloid represented in the general formula (IV), or unsaturated forms thereof and/or seco-, nor- or homo-derivatives thereof:

8. The method of claim 1 or 2, wherein the hedgehog antagonist is a steroidal alkaloid represented in the general formula (V) or unsaturated forms thereof and/or seco-, nor- or homo-derivatives thereof:

9. The method of claim 1 or 2, wherein the hedgehog antagonist is a steroidal alkaloid represented in the general formula (VI), or unsaturated forms thereof and/or seco-, nor- or homo-derivatives thereof:

10. The method of claim 1 or 2, wherein the hedgehog antagonist is a steroidal alkaloid represented in the general formula (VII) or unsaturated forms thereof and/or seco-, nor- or homo-derivatives thereof:

11. The method of any of claims 3-10, 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, triamcinolone and their derivatives.

12. The method of any of claims 3-10, wherein the steroidal alkaloid does not specifically bind a nuclear hormone receptor.

13. The method of any of claims 3-10, wherein the steroidal alkaloid does not specifically bind estrogen or testerone receptors.

14. The method of any of claims 3-10, wherein the steroidal alkaloid has no estrogenic activity at therapeutic concentrations.

15. The method of any of claims 1-10, wherein the hedgehog antagonist inhibits hedgehog-mediated signal transduction with an ED₅₀ of 1 mM or less.

16. The method of any of claims 1-10, wherein the hedgehog antagonist inhibits hedgehog-mediated signal transduction with an ED₅₀ of 1 μ

M or less.

17. The method of any of claims 1-10, wherein the steroidal alkaloid inhibit hedgehog-mediated signal transduction with an ED₅₀ of 1 nM or less.

18. The method of claims 1 or 2, wherein the cell is contacted with the hedgehog antagonist in vitro.

19. The method of claims 1 or 2, wherein the cell is contacted with the hedgehog antagonist in vivo.

20. The method of claim 1 or 2, wherein the hedgehog antagonist is administered as part of a therapeutic or cosmetic application.

21. The method of claim 19 or 20, wherein the hedgehog antagonist is administered to treat a condition selected from the group consisting of regulation of neural tissues, bone and cartilage formation and repair, regulation of spermatogenesis, regulation of smooth muscle, regulation of lung, liver and other organs arising from the primitive gut, regulation of hematopoietic function, regulation of skin and hair growth, etc.

22. The method of any of claims 1-7, wherein the hedgehog antagonist is applied as a topical formulation to skin in order to inhibit aberrant proliferation of epithelial cells.

23. The method of any of claims 1-17 or 22, wherein the hedgehog antagonist is administered to patient to inhibit growth of a basal cell carcinoma.

2. A method for inhibiting an altered growth state of a cell having a ptc loss-of-function phenotype or a smoothened gain-of-function phenotype, comprising contacting the cell with a ptc agonist in a sufficient amount to inhibit the altered growth state, wherein the ptc agonist is a organic molecule having a molecule weight less than 750 amu.

24. A pharmaceutical preparation comprising steroidal alkaloid is represented in the general formulas (I), or unsaturated forms thereof and/or seco-, nor- or homo-derivatives thereof:

25. The preparation of claim 24, formulated for topical application.

26. A process for manufacturing a medicament comprising formulating a steroid alkaloid inhibitor of a hedgehog signal transduction pathway in a pharmaceutically acceptable excipient to from a sterile medicament for preventing growth of cells having an aberrant activation hedgehog pathway.