A Short Review on Introduction and Researches on Anticancerous Activity of Geldanamycin

Hesham Sami*, Rajesh Gour, Akhlesh Kumar Singhai

School of Pharmacy, LNCT University, Kolar Road, Bhopal - 462042, Madhya Pradesh, India.

*Corresponding Author E-mail: heshamsami1999@gmail.com

ABSTRACT:

Geldanamycin (GA) bind heat-shock protein-90 (HSP-90) and destabilize its client proteins including v-Src, Bcr-Abl, RAF-1, Erb-B2, some growth factor receptors and steroid receptors. As a result, several oncoproteins are subjected to ubiquitination and proteasomal destruction by HSP-90 active compounds. HSP-90 active substances can either stop apoptosis from occurring or promote growth arrest, differentiation, and apoptosis depending on the cellular environment. Numerous preclinical models and clinical trials have demonstrated anticancer activity for a number of HSP-90 inhibitors. The well-known HSP-90 inhibitor geldanamycin’s clinical development was hampered by its hepatic toxicity. Geldanamycin at low doses can sensitize Bcr/Abl-expressing leukemia cells to death in the presence of inadequate doxorubicin concentrations by activating caspase. In another example, 17AAG in combination with taxol shows enhanced cytotoxic effects on taxol-resistant Erb-B2 overexpressing breast cancer cells. The benzoquinone ansamycin geldanamycin selectively binds to GRP94 and HSP-90 both in vivo and in vitro. When cells are treated with geldanamycin, HSP-90’s molecular chaperone function is changed. This prevents some cytosolic proteins from maturing, reduces their activity, and/or modifies their stability. On the other hand, nothing is known about GRP94’s function in protein folding or how geldanamycin affects this endoplasmic reticulum (ER) homologue of HSP-90. In this work, we show that geldanamycin is a strong inducer of the cellular stress response in the ER, leading to the transcriptional up-regulation of ER chaperones and production of the gadd153/CHOP transcription factor in a range of cell lines. Here we mention the anticancerous activity of HSP-90 (Heat Shock Protein 90) Inhibitor geldanamycin and some researches in field of anticancerous activity of Geldanamycin.

KEYWORDS: Geldanamycin, Benzoquinone Ansamycin, HSP-90 Inhibitor, Anticancerous, Molecular Chaperone, Uncontrolled Proliferation.

INTRODUCTION:

Geldanamycin is a benzoquinone ansamycin antibiotic that manifests anti-cancer activity through the inhibition of HSP-90 chaperone function. A large range of human malignancies, including melanoma, leukemia, and tumors of the colon, prostate, lung, and breast, exhibit high quantities of the HSP-90 molecular chaperone.

The stability, folding, and activity of HSP-90 associated proteins, or “client proteins,” are thought to be crucially regulated by HSP90 in cancer cells that rely on oncogene proteins that are altered and/or overexpressed. The growth-stimulating proteins and kinases that promote malignant transformation are among these client proteins.¹ Heat shock protein 90 is one of the most abundant cellular proteins. Although its functions are still being characterized it appears to serve as a chaperone for a growing list of cell signaling proteins, including many tyrosine and serine/threonine kinases, involved in proliferation and/or survival.²

Fig. 1: Binding of geldanamycin to HSP-90⁶

HSP-90 functions as a molecular chaperone by binding to various cellular proteins and supporting the proper folding, stability, and function of target proteins. Protein kinases and steroid hormone receptors are only two examples of the many signal-transducing proteins that control cell growth and differentiation that are included in the HSP-90 client proteins. HSP-90 collaborates with other molecular chaperones including Cdc37 and FKBP52 to perform its ATP-dependent activity. Geldanamycin selectively suppresses HSP-90’s vital ATPase activity. Thus, HSP-90 client proteins are destabilized, inactivated, and degraded when cells are treated with geldanamycin. Apoptosis, oncogenesis, growth, survival, and the cell cycle are all regulated by HSP-90 client proteins, which is why geldanamycin inhibits the growth of cancer cells and has anti-cancer properties.³ Geldanamycin a benzoquinone ansamycin, it consists of a quinone ring and a hydrophobic ansa bridge.⁴ A synthetic form of geldanamycin was extracted from Streptomyces hygroscopicus A070101. 11-methoxy-17-formyl-17-demethoxy-18-O-21-O-dihydrogeldanamycin was the structure that was finally determined.⁵

Historical Background: In 1970, DeBoer et al. purified it for the first time using the broth of Streptomyces hygroscopicus var geldanus var nova. Whitesell et al. (1994) were one. The main mechanism of action of GA appears to be interfering with these HSP ability to function.⁴ Geldanamycin has demonstrated strong action in the National Cancer Institute's (NCI) in vitro screen for anticancer drugs, with 50% growth inhibition (GI50) against the most responsive cell lines at concentrations as low as 13nM and a mean GI50 of 180nM against all 60 of the tumor cell lines.⁷ Geldanamycin also active in mouse tumor models as well.⁸ and the National Cancer Institute has chosen it for preclinical study as an anticancer agent. The ability of geldanamycin to deplete cells of two broad classes of growth-regulatory signaling proteins is likely the cause of its antitumor effects: (1) proto-oncogenic protein kinases, which include the nuclear hormone receptor family, which includes the estrogen and androgen hormone receptors and can drive the growth of hormone-dependent cancers of the breast and prostate, and the Erb-B2 and EGF receptor tyrosine kinases, the v-Src family of non-receptor tyrosine kinases, and Raf-1 and CDK4 Ser/Thr kinases. Subsequent immuno precipitation and X-ray crystallographic studies⁹ revealed that after directly binding to HSP-90, geldanamycin prevents HSP-90 multichaperone complexes from forming, which in turn prevents ubiquitin from degrading HSP-90 client proteins. Geldanamycin competes with ATP binding via binding to the conserved N-terminal domain of HSP-90. Additionally, the crystal structure of geldanamycin and HSP-90 demonstrates that this interaction prevents substrate protein binding.¹⁰ Geldanamycin also binds to GRP94, the HSP-90 analogue in the ER¹¹. Even though geldanamycin has strong anti-tumor effects, its severe hepatotoxicity in certain human tumor models caused problems throughout clinical studies.¹² As a result, efforts to find novel classes of HSP-90 inhibitors with reduced toxicity were launched, and the analogue 17AAG(17-allylamino-17-demethoxy-geldanamycin)  as successfully developed. With less toxicity, 17AAG shares all of geldanamycin’s HSP-90 related properties.¹³

Mechanism of action: As a widely expressed, highly conserved 90 kDa molecular chaperone, HSP-90(Heat Shock Protein90) controls the cellular stress response by preserving the shape, stability, and functionality of important client proteins.¹⁴ HSP-90 and its client proteins are necessary for maintaining cellular homeostasis in organisms. In addition, the HSP-90 complex regulates a number of signal transduction pathways and is crucial for the maturation of numerous tumor-promoting client proteins.¹⁵ Expression of HSP-90 is elevated under stress conditions such as heat shock, abnormal pH and nutrient depletion. It is expressed in cancer cells at up to 2- to 10-fold higher levels than found in normal cells.¹⁶ By blocking HSP-90 ATPase activity, geldanamycin breaks up mature multi-chaperone complexes. The liberated client proteins are then broken down by the ubiquitin-proteasome pathway. HSP-90 inhibitor inhibits uncontrolled proliferation, angiogenesis and invasion/ metastasis.¹⁷ The addition of EGFR(epidermal growth factor receptor), CDK4, RAF-1, ARAF, BRAF, AKT (protein kinase Ba), MET (hepatocyte growth factor receptor), PLK1(Polo-like kinase 1), and BCR-ABL, as well as other cancer-relevant client proteins like anti-apoptotic survivin, the catalytic subunit of telomerase hTERT, HIF-1a(hypoxia-inducible factor 1a), and mutated p53, are all induced to degrade when given geldanamycin treatment.¹⁸ Geldanamycin-bound HSP-90 appears to recruit CHIP (carboxy-terminus of HSP-70 interacting protein)¹⁹, which acts as an E3 ubiquitin ligase for many of the client proteins²⁰. CHIP, originally identified as a HSC-70 co-chaperone has both a tetratricopeptide repeat (TPR) motif and a U-box domain. The TPR motif within CHIP interacts with the molecular chaperones HSC-70 and HSP-90, both of which contain the requisite C-terminal EEVD peptide motif, whereas the CHIP U-box domain performs the ubiquitin ligase function. As a consequence of this interaction, HSP-90 client substrates are ubiquitylated and degraded by the 26S proteasome.¹ HSP-90 inhibition is associated with disturbances in the cytoskeleton and cytoskeletal signaling²¹.

Fig. 2: How HSP-90 inhibitor inhibits these kinds of functions.²⁴

When different medications, including geldanamycin and anti-HSP-90 ribozyme, block HSP-90, it makes different cells more vulnerable to complement assault, detergent-induced cell lysis, hypotonic shock, and hypoxia.²² These results provide credence to the notion that HSP-90 plays a crucial role in preserving cellular integrity and indicate that, among other mechanisms, HSP-90 inhibitors work against tumors by making tumor cells more susceptible to different lytic processes.²³

Importance of HSP-90 inhibitor in the treatment of cancer: HSP-90 inhibitors significant contribution to clinical uses. Combination therapy, which combine these medications in low dosages with traditional chemotherapeutic medicines, appear to be a successful strategy for treating a variety of malignancies. For instance, geldanamycin at low doses can sensitize Bcr/Abl-expressing leukemia cells to death in the presence of inadequate doxorubicin concentrations by activating caspase.²⁵ In another example, 17AAG in combination with taxol shows enhanced cytotoxic effects on taxol-resistant Erb-B2 overexpressing breast cancer cells.²⁶ The type and grade of tumor can differentiate the selectivity of drug-induced effects (Clark et al.)²⁷ demonstrated that 17AAG depletes RAF and AKT in human colon cancer cells by inhibiting HSP-90, but does not change the expression of other client proteins. A single injection of a HSP-90 inhibitor, like 17AAG, has been demonstrated to be more effective in specific malignancies for a range of client protein degradation.²⁸

Researches on Geldanamycin:

S. No.	Researcher/ Year	Title of Research	Finding	Ref.
1	Sivahari Prasad Gorantla et al./ 2024	Type II JAK2 Inhibitor CHZ‐868 and HSP-90 Inhibitors Are Potent towards the Ruxolitinib Resistant JAK2 Variants in JAK2 Driven Myeloproliferative Neoplasms.	HSP-90 inhibitors are potent against ruxolitinib resistant variants through the JAK2 degradation and provides the rationale for clinical evaluation of potent HSP90 inhibitors in genetic resistance driven by JAK2 inhibitors.	29
2	M Lizeth Orozco Morales et al. / 2023	Geldanamycin treatment does not result in anti-cancer activity in a preclinical model of orthotopic mesothelioma.	Geldanamycin as a single agent does not appear to be a viable treatment for mesothelioma.	30
3	Wenwen Yi et al. / 2022	Cytotoxic metabolites from the marine-associated Streptomyces sp. ZZ1944.	Marine-derived actinomycetes from the genus Streptomycete have a huge potential for the production of metabolites one of them is Seco-Geldanamycin A.	31
4	Tipparat Samsawat et al. / 2021	Evaluating the effect of amine-geldanamycin hybrids on anticancer activity.	Three new geldanamycin (GDM) derivatives, 17-((S)-2-amino-3-(1H-indol-3-ylpropan-1-ol)-17-demethoxygeldanamycin (2), 17-((S)-2-amino-3-phenylpropan-1-ol)-17-demethoxygeldanamycin (3), and 17-((S)-4-(2-amino-3-hydroxypropyl) phenol)-17-demethoxygeldanamycin (4), were synthesized by nucleophilic substitution of GDM (1) has also anticancerous activity.	32
5	Ean-Jeong Seo et al. / 2020	Molecular determinants of the response of cancer cells towards geldanamycin and its derivatives	17-desmethoxy-17-N,N-dimethylamino-geldanamycin (17-DMAG) is the compound with the highest binding affinity (−7.73 ± 0.12 kcal/mol) and the lowest inhibition constant (2.16 ± 0.49 μM)	33
6	Thongchai Taechowisan et al / 2019	Anti-Inflammatory activity of geldanamycin and its derivatives in LPS-induced RAW 264.7 cells	Two new gelda-namycin derivatives 17-(tryptamine)-17-demethoxygeldanamycin (2) and 17-(5’-methoxytryptamine)-17-demethoxygeldanamycin (3) possess anti-inflammatory activity on LPS-induced RAW 264.7 cells.	34
7	Hassan Mellatyar et al / 2018	17-DMAG-loaded nanofibrous scaffold for effective growth inhibition of lung cancer cells through targeting HSP-90 gene expression	The implantable 17-DMAG-loaded nanofibrous scaffolds might be an excellent tool for efficiently killing of the lung residual cancer cells and avoid the local cancer recurrence.	35
8	Xinran Wang et al. / 2017	Improved PKS gene expression with strong endogenous promoter resulted in geldanamycin yield increase.	Combined overexpression of the 6‐gene AHBA biosynthetic cassette and PKS genes increased the yield of geldanamycin by 88%, from 773 mg/L of the wild‐type to 1450 mg/L in the derived strain.	36
9	Qiang Huo et al. / 2016	Biosynthesis of novel glucosides geldanamycin analogs by enzymatic synthesis.	Two new glucosides (1 and 2) of geldanamycin (GA) analogs were obtained from in vitro glycosylation by UDP-glycosyltransferase (YjiC) 4, 5-dihydro-7-O-descarbamoyl-7-hydroxyl GA-7-O-β-D-glucoside (1) and ACDL3172-18-O-β-D-glucoside (2).	37
10	Yan-ping Li et al / 2015	Synthesis and biological evaluation of geldanamycin analogs against human cancer cells.	The anti-proliferation activity of 19-methylthio-substituted geldanamycins was significantly lower compared with no 19-substitution geldanamycins in all tested cancer cells.	38

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Received on 05.04.2024         Modified on 08.07.2024

Accepted on 16.08.2024  ©Asian Pharma Press All Right Reserved

Asian J. Pharm. Tech. 2024; 14(3):220-224.

DOI: 10.52711/2231-5713.2024.00036