---
title: "Pyrazolo[3,4-b]pyridine chemotype as a dual FAK1/FAK2 inhibitor for triple-negative breast cancer"
id: "pubmed-42172799"
canonical_url: "https://medichelpline.com/clinical-feed/pubmed-42172799"
content_type: "clinical_feed_article"
specialty: "Oncology"
source_name: "PubMed / NCBI"
source_url: "https://pubmed.ncbi.nlm.nih.gov/42172799/"
doi: "10.1016/j.bioorg.2026.110005"
published_at: "2026-09-05T00:00:00.000Z"
evidence_level: "Journal Article"
license: "CC-BY-NC-4.0 / Informational Use"
---
# Pyrazolo[3,4-b]pyridine chemotype as a dual FAK1/FAK2 inhibitor for triple-negative breast cancer
## Provenance & Clinical Metadata
- **Canonical URL:** https://medichelpline.com/clinical-feed/pubmed-42172799
- **Specialty:** [Oncology](https://medichelpline.com/clinical-feed/oncology.md)
- **Primary Source:** PubMed / NCBI
- **Source URL:** [Original Journal Publication](https://pubmed.ncbi.nlm.nih.gov/42172799/)
- **DOI:** [10.1016/j.bioorg.2026.110005](https://doi.org/10.1016%2Fj.bioorg.2026.110005)
- **Published At:** 2026-09-05T00:00:00.000Z
- **Evidence Rating:** Journal Article
## Executive GIST (TL;DR)
- Triple-negative breast cancer (TNBC) has limited therapeutic targets and remains difficult to treat, prompting interest in new targeted agents. - The study used virtual screening to identify a **pyrazolo[3,4-b]pyridine** scaffold as a candidate for dual inhibition of **FAK1/FAK2**. - Sixteen derivatives of the scaffold were synthesized and evaluated; compound **8a** emerged with the strongest binding to both FAK1 and FAK2. - Measured KD values for 8a were 2.9 μM (FAK1) and 2.5 μM (FAK2), indicating micromolar affinity for both kinases. - Intracellular target engagement assays showed that 8a protected specific peptides within the kinase domains of FAK1 and FAK2 inside cells, supporting on-target binding. - Functionally, 8a caused G2/M cell-cycle arrest and activated DNA damage–related signaling, with a reduction in CDK1 phosphorylation. - The antiproliferative effect of 8a was reduced when FAK1/FAK2 were knocked down, indicating target-dependence. - Combining 8a with doxorubicin produced a significantly enhanced antiproliferative effect compared with 8a alone. - In vivo, 8a inhibited growth of 4 T1 tumors without causing significant body weight loss in treated animals. - The authors conclude that the **pyrazolo[3,4-b]pyridine** scaffold is a novel chemotype for **dual FAK1/FAK2** targeting and that 8a is a target-dependent lead for anti-TNBC development. - Specific experimental details (full structure-activity relationships, dosing regimens, statistical analyses, and comprehensive safety data) were not reported in the abstract and require consultation of the full text for verification.
## Clinical Analysis & Structured Key Points
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Epub 2026 May 19. # Identification of a pyrazolo[3,4-b]pyridine chemotype for dual focal adhesion kinase 1/2 (FAK1/2) targeting in triple-negative breast cancer [Huirong Li](https://pubmed.ncbi.nlm.nih.gov/?term=Li+H&cauthor_id=42172799)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42172799/#full-view-affiliation-1 "School of Pharmacy, Key Laboratory of Molecular Pharmacology and Drug Evaluation \(Yantai University\), Ministry of Education, Basic Science Research Center Base \(Pharmaceutical Science\), Yantai University, Yantai 264005, China."), [Lili Sun](https://pubmed.ncbi.nlm.nih.gov/?term=Sun+L&cauthor_id=42172799)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42172799/#full-view-affiliation-1 "School of Pharmacy, Key Laboratory of Molecular Pharmacology and Drug Evaluation \(Yantai University\), Ministry of Education, Basic Science Research Center Base \(Pharmaceutical Science\), Yantai University, Yantai 264005, China."), [Huilin Ling](https://pubmed.ncbi.nlm.nih.gov/?term=Ling+H&cauthor_id=42172799)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42172799/#full-view-affiliation-1 "School of Pharmacy, Key Laboratory of Molecular Pharmacology and Drug Evaluation \(Yantai University\), Ministry of Education, Basic Science Research Center Base \(Pharmaceutical Science\), Yantai University, Yantai 264005, China."), [Yanfei Chang](https://pubmed.ncbi.nlm.nih.gov/?term=Chang+Y&cauthor_id=42172799)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42172799/#full-view-affiliation-1 "School of Pharmacy, Key Laboratory of Molecular Pharmacology and Drug Evaluation \(Yantai University\), Ministry of Education, Basic Science Research Center Base \(Pharmaceutical Science\), Yantai University, Yantai 264005, China."), [Shuo Ren](https://pubmed.ncbi.nlm.nih.gov/?term=Ren+S&cauthor_id=42172799)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42172799/#full-view-affiliation-1 "School of Pharmacy, Key Laboratory of Molecular Pharmacology and Drug Evaluation \(Yantai University\), Ministry of Education, Basic Science Research Center Base \(Pharmaceutical Science\), Yantai University, Yantai 264005, China."), [Huicong Xin](https://pubmed.ncbi.nlm.nih.gov/?term=Xin+H&cauthor_id=42172799)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42172799/#full-view-affiliation-1 "School of Pharmacy, Key Laboratory of Molecular Pharmacology and Drug Evaluation \(Yantai University\), Ministry of Education, Basic Science Research Center Base \(Pharmaceutical Science\), Yantai University, Yantai 264005, China."), [Hongbo Wang](https://pubmed.ncbi.nlm.nih.gov/?term=Wang+H&cauthor_id=42172799)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42172799/#full-view-affiliation-1 "School of Pharmacy, Key Laboratory of Molecular Pharmacology and Drug Evaluation \(Yantai University\), Ministry of Education, Basic Science Research Center Base \(Pharmaceutical Science\), Yantai University, Yantai 264005, China."), [Qikun Yin](https://pubmed.ncbi.nlm.nih.gov/?term=Yin+Q&cauthor_id=42172799)[ 2 ](https://pubmed.ncbi.nlm.nih.gov/42172799/#full-view-affiliation-2 "School of Pharmacy, Key Laboratory of Molecular Pharmacology and Drug Evaluation \(Yantai University\), Ministry of Education, Basic Science Research Center Base \(Pharmaceutical Science\), Yantai University, Yantai 264005, China.. Electronic address: qkyin@ytu.edu.cn.") Affiliations Expand ### Affiliations * 1 School of Pharmacy, Key Laboratory of Molecular Pharmacology and Drug Evaluation (Yantai University), Ministry of Education, Basic Science Research Center Base (Pharmaceutical Science), Yantai University, Yantai 264005, China. * 2 School of Pharmacy, Key Laboratory of Molecular Pharmacology and Drug Evaluation (Yantai University), Ministry of Education, Basic Science Research Center Base (Pharmaceutical Science), Yantai University, Yantai 264005, China.. Electronic address: qkyin@ytu.edu.cn. * PMID: **42172799** * DOI: [ 10.1016/j.bioorg.2026.110005 ](https://doi.org/10.1016/j.bioorg.2026.110005) Item in Clipboard # Identification of a pyrazolo[3,4-b]pyridine chemotype for dual focal adhesion kinase 1/2 (FAK1/2) targeting in triple-negative breast cancer Huirong Li et al. Bioorg Chem. 2026. Show details Display options Display options Format Abstract PubMed PMID Bioorg Chem Actions * [ Search in PubMed ](https://pubmed.ncbi.nlm.nih.gov/?term=%22Bioorg+Chem%22%5Bjour%5D&sort=date&sort_order=desc) * [ Search in NLM Catalog ](https://www.ncbi.nlm.nih.gov/nlmcatalog?term=%22Bioorg+Chem%22%5BTitle+Abbreviation%5D) * [ Add to Search ](https://pubmed.ncbi.nlm.nih.gov/42172799/) . 2026 Sep 5:179:110005. doi: 10.1016/j.bioorg.2026.110005. Epub 2026 May 19. ### Authors [Huirong Li](https://pubmed.ncbi.nlm.nih.gov/?term=Li+H&cauthor_id=42172799)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42172799/#short-view-affiliation-1 "School of Pharmacy, Key Laboratory of Molecular Pharmacology and Drug Evaluation \(Yantai University\), Ministry of Education, Basic Science Research Center Base \(Pharmaceutical Science\), Yantai University, Yantai 264005, China."), [Lili Sun](https://pubmed.ncbi.nlm.nih.gov/?term=Sun+L&cauthor_id=42172799)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42172799/#short-view-affiliation-1 "School of Pharmacy, Key Laboratory of Molecular Pharmacology and Drug Evaluation \(Yantai University\), Ministry of Education, Basic Science Research Center Base \(Pharmaceutical Science\), Yantai University, Yantai 264005, China."), [Huilin Ling](https://pubmed.ncbi.nlm.nih.gov/?term=Ling+H&cauthor_id=42172799)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42172799/#short-view-affiliation-1 "School of Pharmacy, Key Laboratory of Molecular Pharmacology and Drug Evaluation \(Yantai University\), Ministry of Education, Basic Science Research Center Base \(Pharmaceutical Science\), Yantai University, Yantai 264005, China."), [Yanfei Chang](https://pubmed.ncbi.nlm.nih.gov/?term=Chang+Y&cauthor_id=42172799)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42172799/#short-view-affiliation-1 "School of Pharmacy, Key Laboratory of Molecular Pharmacology and Drug Evaluation \(Yantai University\), Ministry of Education, Basic Science Research Center Base \(Pharmaceutical Science\), Yantai University, Yantai 264005, China."), [Shuo Ren](https://pubmed.ncbi.nlm.nih.gov/?term=Ren+S&cauthor_id=42172799)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42172799/#short-view-affiliation-1 "School of Pharmacy, Key Laboratory of Molecular Pharmacology and Drug Evaluation \(Yantai University\), Ministry of Education, Basic Science Research Center Base \(Pharmaceutical Science\), Yantai University, Yantai 264005, China."), [Huicong Xin](https://pubmed.ncbi.nlm.nih.gov/?term=Xin+H&cauthor_id=42172799)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42172799/#short-view-affiliation-1 "School of Pharmacy, Key Laboratory of Molecular Pharmacology and Drug Evaluation \(Yantai University\), Ministry of Education, Basic Science Research Center Base \(Pharmaceutical Science\), Yantai University, Yantai 264005, China."), [Hongbo Wang](https://pubmed.ncbi.nlm.nih.gov/?term=Wang+H&cauthor_id=42172799)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42172799/#short-view-affiliation-1 "School of Pharmacy, Key Laboratory of Molecular Pharmacology and Drug Evaluation \(Yantai University\), Ministry of Education, Basic Science Research Center Base \(Pharmaceutical Science\), Yantai University, Yantai 264005, China."), [Qikun Yin](https://pubmed.ncbi.nlm.nih.gov/?term=Yin+Q&cauthor_id=42172799)[ 2 ](https://pubmed.ncbi.nlm.nih.gov/42172799/#short-view-affiliation-2 "School of Pharmacy, Key Laboratory of Molecular Pharmacology and Drug Evaluation \(Yantai University\), Ministry of Education, Basic Science Research Center Base \(Pharmaceutical Science\), Yantai University, Yantai 264005, China.. Electronic address: qkyin@ytu.edu.cn.") ### Affiliations * 1 School of Pharmacy, Key Laboratory of Molecular Pharmacology and Drug Evaluation (Yantai University), Ministry of Education, Basic Science Research Center Base (Pharmaceutical Science), Yantai University, Yantai 264005, China. * 2 School of Pharmacy, Key Laboratory of Molecular Pharmacology and Drug Evaluation (Yantai University), Ministry of Education, Basic Science Research Center Base (Pharmaceutical Science), Yantai University, Yantai 264005, China.. Electronic address: qkyin@ytu.edu.cn. * PMID: **42172799** * DOI: [ 10.1016/j.bioorg.2026.110005 ](https://doi.org/10.1016/j.bioorg.2026.110005) Item in Clipboard Full text links Cite Display options Display options Format Abstract PubMed PMID ## Abstract Triple-negative breast cancer (TNBC) remains difficult to treat because of the lack of effective therapeutic targets and limited treatment options. Dual targeting of Focal adhesion kinase 1 (FAK1) and 2 (FAK2) has emerged as a promising strategy, yet current dual-targeted ligands are still limited in structural diversity and binding mode. In this study, a pyrazolo[3,4-b]pyridine scaffold was identified through virtual screening and optimized to generate 16 derivatives. Among them, compound 8a showed the strongest binding affinity for both FAK1 and FAK2, with KD values of 2.9 μM and 2.5 μM, respectively, and exhibited the best selectivity toward TNBC cells. Intracellular target engagement studies indicated that 8a protected specific peptides within the kinase domains of FAK1 and FAK2 in cells. Functionally, 8a induced G2/M-phase arrest and activated DNA damage-related signaling, accompanied by decreased phosphorylation of CDK1. The antiproliferative activity of 8a was markedly attenuated by FAK1/FAK2 knockdown but significantly enhanced in combined treatment with doxorubicin. In addition, 8a inhibited 4 T1 tumor growth in vivo without significant body weight loss. These findings identify pyrazolo[3,4-b]pyridine as a new chemotype for dual FAK1/FAK2 targeting and support 8a as a target-dependent lead compound for anti-TNBC development. **Keywords:** Dual-target ligand; Focal adhesion kinase; Pyrazolo[3,4-b]pyridine chemotype; Structure-activity relationship; Triple-negative breast cancer. Copyright © 2026 Elsevier Inc. All rights reserved. [PubMed Disclaimer](https://pubmed.ncbi.nlm.nih.gov/disclaimer/) ## Conflict of interest statement Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. ## MeSH terms * Animals Actions * [ Search in PubMed ](https://pubmed.ncbi.nlm.nih.gov/?term=%22Animals%22%5BMeSH%5D&sort=date&sort_order=desc) * [ Search in MeSH ](https://www.ncbi.nlm.nih.gov/mesh?term=Animals) * [ Add to Search ](https://pubmed.ncbi.nlm.nih.gov/42172799/) * Antineoplastic Agents* / chemical synthesis Actions * [ Search in PubMed ](https://pubmed.ncbi.nlm.nih.gov/?term=%22Antineoplastic+Agents%2Fchemical+synthesis%22%5BMAJR%5D&sort=date&sort_order=desc) * [ Search in MeSH ](https://www.ncbi.nlm.nih.gov/mesh?term=Antineoplastic+Agents) * [ Add to Search ](https://pubmed.ncbi.nlm.nih.gov/42172799/) * Antineoplastic Agents* / chemistry Actions * [ Search in PubMed ](https://pubmed.ncbi.nlm.nih.gov/?term=%22Antineoplastic+Agents%2Fchemistry%22%5BMAJR%5D&sort=date&sort_order=desc) * [ Search in MeSH ](https://www.ncbi.nlm.nih.gov/mesh?term=Antineoplastic+Agents) * [ Add to Search ](https://pubmed.ncbi.nlm.nih.gov/42172799/) * Antineoplastic Agents* / pharmacology Actions * [ Search in PubMed ](https://pubmed.ncbi.nlm.nih.gov/?term=%22Antineoplastic+Agents%2Fpharmacology%22%5BMAJR%5D&sort=date&sort_order=desc) * [ Search in MeSH ](https://www.ncbi.nlm.nih.gov/mesh?term=Antineoplastic+Agents) * [ Add to Search ](https://pubmed.ncbi.nlm.nih.gov/42172799/) * Cell Line, Tumor Actions * [ Search in PubMed ](https://pubmed.ncbi.nlm.nih.gov/?term=%22Cell+Line%2C+Tumor%22%5BMeSH%5D&sort=date&sort_order=desc) * [ Search in MeSH ](https://www.ncbi.nlm.nih.gov/mesh?term=Cell+Line%2C+Tumor) * [ Add to Search ](https://pubmed.ncbi.nlm.nih.gov/42172799/) * Cell Proliferation / drug effects Actions * [ Search in PubMed ](https://pubmed.ncbi.nlm.nih.gov/?term=%22Cell+Proliferation%2Fdrug+effects%22%5BMeSH%5D&sort=date&sort_order=desc) * [ Search in MeSH ](https://www.ncbi.nlm.nih.gov/mesh?term=Cell+Proliferation) * [ Add to Search ](https://pubmed.ncbi.nlm.nih.gov/42172799/) * Dose-Response Relationship, Drug Actions * [ Search in PubMed ](https://pubmed.ncbi.nlm.nih.gov/?term=%22Dose-Response+Relationship%2C+Drug%22%5BMeSH%5D&sort=date&sort_order=desc) * [ Search in MeSH ](https://www.ncbi.nlm.nih.gov/mesh?term=Dose-Response+Relationship%2C+Drug) * [ Add to Search ](https://pubmed.ncbi.nlm.nih.gov/42172799/) * Drug Screening Assays, Antitumor Actions * [ Search in PubMed ](https://pubmed.ncbi.nlm.nih.gov/?term=%22Drug+Screening+Assays%2C+Antitumor%22%5BMeSH%5D&sort=date&sort_order=desc) * [ Search in MeSH ](https://www.ncbi.nlm.nih.gov/mesh?term=Drug+Screening+Assays%2C+Antitumor) * [ Add to Search ](https://pubmed.ncbi.nlm.nih.gov/42172799/) * Female Actions * [ Search in PubMed ](https://pubmed.ncbi.nlm.nih.gov/?term=%22Female%22%5BMeSH%5D&sort=date&sort_order=desc) * [ Search in MeSH ](https://www.ncbi.nlm.nih.gov/mesh?term=Female) * [ Add to Search ](https://pubmed.ncbi.nlm.nih.gov/42172799/) * Focal Adhesion Kinase 1* / antagonists & inhibitors Actions * [ Search in PubMed ](https://pubmed.ncbi.nlm.nih.gov/?term=%22Focal+Adhesion+Kinase+1%2Fantagonists+and+inhibitors%22%5BMAJR%5D&sort=date&sort_order=desc) * [ Search in MeSH
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