US-12617757-B2 - Compounds and modules for inhibition of pre-miR-21 and their use in treatment of certain cancers

Abstract

Small molecule compounds and corresponding dimers having inhibitory activity against pre-miR-21 RNA and related methods for treatment of neoplastic disease such as cancer and especially cancers expressing miR-21 are disclosed.

Inventors

• Matthew David Disney
• Jessica L. Childs-Disney

Assignees

• UNIVERSITY OF FLORIDA RESEARCH FOUNDATION, INCORPORATED

Dates

Publication Date

20260505

Application Date

20201029

Claims (6)

1. 1 . A compound of Formula 5 and the pharmaceutically acceptable salts thereof wherein designator m is 4, and each R 8 is independently hydrogen or methyl.
2. 2 . A pharmaceutical composition comprising a pharmaceutically acceptable carrier and an amount of a compound of claim 1 , or a pharmaceutically acceptable salt thereof, which is effective for inhibition of RNA pre-miR-21 activity.
3. 3 . A method for decreasing RNA pre-miR-21 activity in a patient, for treatment of cancer in a patient wherein the cancer cells of the patient display miR-21 RNA activity and/or pre-miR-21 expression, or for treatment of a patient's disease mediated by aberrant miR-21 RNA activity, comprising administering to the patient an effective amount of a pharmaceutical composition of claim 2 .
4. 4 . The method of claim 3 , wherein the cancer is breast, lung, pancreatic or melanoma cancer.
5. 5 . A method of cellular destruction by targeting an oncogenic non-coding RNA precursor, comprising contacting a cell expressing the oncogenic non-coding RNA precursor with a pharmaceutical composition of claim 2 .
6. 6 . The method of claim 5 wherein the oncogenic non-coding RNA precursor comprises oncogenic pre-microRNA-21 (pre-miR-21).

Description

CROSS-REFERENCE TO RELATED APPLICATION This application is a U.S. national stage filing under 35 U.S.C. § 371 from International Application No. PCT/US2020/057914, filed on 29 Oct. 2020, and published as WO2021/087084 on 6 May 2021, which claims the priority of U.S. provisional application Ser. No. 62/927,247, filed Oct. 29, 2019, the benefit of priority of each of which is claimed herein, and which applications and publication are hereby incorporated by reference in their entirety. STATEMENT OF GOVERNMENT SUPPORT This invention was made with Government support under R01GM097455-09 awarded by the National Institutes of Health. The Government has certain rights in this invention. REFERENCE TO AN ELECTRONIC SEQUENCE LISTING The contents of the electronic sequence listing (U120270138US01-SEQ-CLL.txt; Size: 7,417 bytes; and Date of Creation: Sep. 23, 2025) are herein incorporated by reference in its entirety. BACKGROUND RNA is involved with a myriad of cellular roles beyond merely encoding and assembling proteins. The Encyclopedia of DNA Elements project and subsequent analyses showed that only 1-2% of our genome encodes for protein yet about 80% of it is transcribed into RNA (ENCODE, 2012). Although the majority of transcribed RNAs are non-coding, many non-coding RNAs are functionally involved in modulating cell activities and disease states. Such functional, non-coding RNAs represent a potential therapeutic target. Indeed, RNA structures have been shown to be key players in important biological processes and in diseased states. Because RNA biology is often mediated by the structures that it forms, approaches to target structured RNAs could be advantageous. Currently, the approach to target RNA has been the use of oligonucleotide compounds that preferentially target unstructured regions1 Thus, the development of therapeutics that target RNA has mostly centered on using oligonucleotides. Small molecules interacting with a RNA's three-dimensional structure could allow one to preferentially target RNA structure. However, interactions between RNA and small molecule compounds are poorly understood which has led to the perception that RNA is “undruggable”. (Guan & Disney, 2012; Thomas & Hergenrother, 2008). Nevertheless, small molecules have shown an ability to target RNA. For example, small molecules have been investigated for targeting the three-dimensional structure of ribosome, riboswitches, certain viral RNA and nucleotide repeat expansions. (Blount and Breaker, 2006; U.S. Pat. No. 9,586,944 B2; U.S. Patent Application Publication No. 2016/0188791 A1). One class of structured RNAs that play roles in disease biology are non-coding microRNAs (miRs). They are produced from highly structured precursors that are processed in the nucleus (pri-miRs) and cytoplasm (pre-miRs) by the nucleases Drosha and Dicer, respectively see FIGS. 1A-1B. Many miRs play significant roles in human disease biology, see D. P. Bartel, MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 116, 281-297 (2004). For example, miR-21's expression negatively correlates with survival in triple negative breast cancer patients and is expressed in solid tumors, see A. M. Krichevsky, G. Gabriely, miR-21: a small multi-faceted RNA. J. Cell. Mol. Med. 13, 39-53 (2009). Therefore, an object of the present invention is the development of small molecules that achieve modification, amelioration and/or negation of miR-21 activity. Another object of the present invention is the development of small molecules that selectively inhibit pre-miR-21 so as to silence, modify and/or ameliorate the expression of miR-21. Yet another object is to target pre-miR-21 for enzymatic cleavage. A further object is to develop an inhibitory and degradation small molecule module that displays high binding and selectivity targeting of pre-miR-21. SUMMARY OF THE INVENTION These and other objects are achieved by embodiments of the present invention that are directed to a small molecule compound comprising Formula 1 and the pharmaceutically acceptable salts thereof. For Formula 1, R8 is hydrogen or methyl, R9 is hydroxyl, H2N—(CH2)q—NH— or N3—(CH2)q—NH—, designator q is an integer of 2 to 6 and designator r is an integer of 2 to 6. Preferably, R8 is methyl. Embodiments of Formula 1 including those when R9 is hydroxyl or α, ω-diaminoalkylenyl, or N3—(CH2)q—NH— display inhibitory activity against expression of the structured RNA pre-miR-21. The embodiments of Formula 1 and especially those comprising R9 as the azidoalkylamino group, N3—(CH2)q—NH— display inhibition of the expression of RNA pre-miR-21 so that production of miR-21 is ameliorated, curtailed, minimized and/or silenced. The interaction of embodiments of Formula 1 with pre-miR-21 enable treatment of diseases associated with expression of miR-21, such as neoplastic disease, cancer and especially lung and breast cancer. The inhibition of the expression of miR-21 and associated nuclease degradation leads to programmed