EP0629697A2
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EP
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Materials and methods for screening anti-osteoporosis agents
Assignee: Eli Lilly and Co
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Filing: 1994-06-20
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Granted: —
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Expires: 2014-06-20
Abstract
Abstract The present invention relates to methods for the identification of therapeutic agents for the treatment of osteoporosis and serum lipid lowering agents. The invention relates to isolating cloning, and using nucleic acids from the promoter regions of transforming growth factor beta genes comprising novel regulatory elements designated "raloxifene responsive elements". The invention also encompasses eukaryotic cells containing such raloxifene responsive elements operably linked to reporter genes such that the raloxifene responsive elements modulate the transcription of the reporter genes. The invention provides methods for identifying anti-osteoporosis agents that induce transcription of genes operably linked to such raloxifene responsive elements without inducing deleterious or undesirable side effects associated with current anti-osteoporosis therapy regimens.
Inventors
- 👤 Na Nora Yang
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-23 21:17
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/79; C12N15/85; A; A61; A61K; A61K31/00; A61K31/33; A61K31/38; A61K31/381; A61K31/395; A61K31/40; A61K31/435; A61K31/44; A61K31/445; C07; C07K; C07K14/00; C07K14/435; C07K14/46; C07K14/475; C07K14/495; C07K14/705; C07K14/72; C07K14/721; C12Q; C12Q1/00; C12Q1/68; C12Q1/6897; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/68; G01N33/6893; C12N2830/00; C12N2830/001; C12N2830/002; C12N2830/008; C12N2830/80; C12N2830/85; G01N2333/00; G01N2333/435; G01N2333/475; G01N2333/495; G01N2500/00; G01N2500/04
Claims Analysis
Total Claims: 10
Claim 1
Independent Claim
26 words
A nucleic acid comprising a raloxifene responsive element isolated from a promoter region of a TGFβ gene wherein the human TGFβ gene is TGFβ-2 or TGFβ-3.
Claim 2
Dependent Claim
137 words
A nucleic acid according to Claim 1 wherein the nucleotide sequence of the nucleic acid comprises a sequence selected from the group consisting of sequences corresponding to 1896 to 2306 shown in Figure 3 and SEQ ID NO:3, sequences corresponding to 1976 to 2306 shown in Figure 3 and SEQ ID NO:3, sequences corresponding to 2106 to 2306 shown in Figure 3 and SEQ ID NO:3, sequences corresponding to 2137 to 2306 shown in Figure 3 and SEQ ID NO:3, sequences corresponding to 2150 to 2306 shown in Figure 3 and SEQ ID NO:3, sequences corresponding to 2159 to 2306 shown in Figure 3 and SEQ ID NO:3, sequences corresponding to 2159 to 2271 shown in Figure 3 and SEQ ID NO:3, and sequences corresponding to 2159 to 2231 shown in Figure 3 and SEQ ID NO:3.
Claim 3
Dependent Claim
19 words
A recombinant expression construct comprising the nucleic acid according to Claim 1 or Claim 2 and a reporter gene.
Claim 4
Dependent Claim
13 words
A eukaryotic cell transfected with the recombinant expression construct according to Claim 3.
Claim 5
Independent Claim
157 words
A method for screening a multiplicity of compounds to identify compounds having potential as anti-osteoporosis agents, the method comprising identifying a compound of the multiplicity that is capable of inducing transcription from a raloxifene responsive element of a mammalian promoter, that is not a non-specific transcription inducer, is not capable of inducing transcription from an estrogen-responsive element of a mammalian promoter and that is an anti-estrogenic or non-estrogenic compound, the method comprising the steps of: (a) assaying for the ability of the compound to induce transcription from a raloxifene responsive element of the mammalian promoter; (b) assaying for the inability of the compound to induce transcription from a mammalian promoter not having a raloxifene responsive element; (c) assaying for the inability of the compound to induce transcription from an estrogen responsive promoter; and (d) assaying for the ability of the compound to inhibit estrogen induction of transcription from an estrogen responsive promoter in the presence of estrogen.
Claim 6
Independent Claim
108 words
A method for inducing bone formation in a mammal which comprises administering a compound that, when bound to an estrogen receptor, potently induces transcription from a raloxifene responsive element of a promoter region of a TGFβ-3 gene, provided the compound is other than a compound of the formula wherein n is 0, 1 or 2; R and R¹, independently, are hydrogen, hydroxyl, C₁-C₆-alkoxy, C₁-C₆-acyloxy, C₁-C₆-alkoxy-C₂-C₆-acyloxy, R³-substituted aryloxy, R³-substituted aroyloxy, R⁴-substituted carbonyloxy, chloro, or bromo; R² is a heterocyclic ring selected from the group consisting of pyrrolidino, piperidino, or hexamethyleneimino; R³ is C₁-C₃-alkyl, C₁-C₃-alkoxy, hydrogen, or halo; and R⁴ is C₁-C₆-alkoxy or aryloxy; or a pharmaceutically acceptable salt thereof.
Claim 7
Independent Claim
104 words
A method for treating osteoporosis which comprises administering a compound that, when bound to an estrogen receptor, potently induces transcription from a raloxifene responsive element of a promoter region of a TGFβ-3 gene, provided the compound is other than a compound of the formula wherein n is 0, 1 or 2; R and R¹, independently, are hydrogen, hydroxyl, C₁-C₆-alkoxy, C₁-C₆-acyloxy, C₁-C₆-alkoxy-C₂-C₆-acyloxy, R³-substituted aryloxy, R³-substituted aroyloxy, R⁴-substituted carbonyloxy, chloro, or bromo; R² is a heterocyclic ring selected from the group consisting of pyrrolidino, piperidino, or hexamethyleneimino; R³ is C₁-C₃-alkyl, C₁-C₃-alkoxy, hydrogen, or halo; and R⁴ is C₁-C₆-alkoxy or aryloxy; or a pharmaceutically acceptable salt thereof.
Claim 8
Independent Claim
105 words
A method for treating bone fractures which comprises administering a compound that, when bound to an estrogen receptor, potently induces transcription from a raloxifene responsive element of a promoter region of a TGFβ-3 gene, provided the compound is other than a compound of the formula wherein n is 0, 1 or 2; R and R¹, independently, are hydrogen, hydroxyl, C₁-C₆-alkoxy, C₁-C₆-acyloxy, C₁-C₆-alkoxy-C₂-C₆-acyloxy, R³-substituted aryloxy, R³-substituted aroyloxy, R⁴-substituted carbonyloxy, chloro, or bromo; R² is a heterocyclic ring selected from the group consisting of pyrrolidino, piperidino, or hexamethyleneimino; R³ is C₁-C₃-alkyl, C₁-C₃-alkoxy, hydrogen, or halo; and R⁴ is C₁-C₆-alkoxy or aryloxy; or a pharmaceutically acceptable salt thereof.
Claim 9
Independent Claim
143 words
A method for inducing bone formation in a mammal which comprises administering a compound that: (a) potently induces transcription from a raloxifene responsive element of a promoter region of a TGFβ-3 gene; (b) does not induce transcription from a mammalian promoter not having a raloxifene responsive element; (c) does not induce transcription from an estrogen responsive promoter; and (d) inhibits estrogen-induced transcription from an estrogen responsive promoter in the presence of estrogen; provided the compound is other than a compound of the formula wherein n is 0, 1 or 2; R and R¹, independently, are hydrogen, hydroxyl, C₁-C₆-alkoxy, C₁-C₆-acyloxy, C₁-C₆-alkoxy-C₂-C₆-acyloxy, R³-substituted aryloxy, R³-substituted aroyloxy, R⁴-substituted carbonyloxy, chloro, or bromo; R² is a heterocyclic ring selected from the group consisting of pyrrolidino, piperidino, or hexamethyleneimino; R³ is C₁-C₃-alkyl, C₁-C₃-alkoxy, hydrogen, or halo; and R⁴ is C₁-C₆-alkoxy or aryloxy; or a pharmaceutically acceptable salt thereof.
Claim 10
Dependent Claim
109 words
The use of a compound that, when bound to an estrogen receptor, potently induces transcription from a raloxifene responsive element of a promoter region of a TGFβ-3 gene, provided the compound is other than a compound of the formula wherein n is 0, 1 or 2; R and R¹, independently, are hydrogen, hydroxyl, C₁-C₆-alkoxy, C₁-C₆-acyloxy, C₁-C₆-alkoxy-C₂-C₆-acyloxy, R³-substituted aryloxy, R³-substituted aroyloxy, R⁴-substituted carbonyloxy, chloro, or bromo; R² is a heterocyclic ring selected from the group consisting of pyrrolidino, piperidino, or hexamethyleneimino; R³ is C₁-C₃-alkyl, C₁-C₃-alkoxy, hydrogen, or halo; and R⁴ is C₁-C₆-alkoxy or aryloxy; or a pharmaceutically acceptable salt thereof; in the preparation of a medicament useful for treating osteoporosis.
References Cited (Backward Citations)
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