Co-delivery of Cu(I) chelator and chemotherapeutics as a new strategy for tumor theranostic

doi:10.1016/j.jconrel.2020.02.023

Journal of Controlled Release

Volume 321, 10 May 2020, Pages 483-496

https://doi.org/10.1016/j.jconrel.2020.02.023 Get rights and content

Highlights

•
A theranostic specific Cu⁺ chelator was proposed.
•
Targeting micelle co-loaded with Cu⁺-chelator and chemotherapeutic reagents was fabricated.
•
Antiangiogenesis and tumor's oxygen saturation reduction were reduced.
•
Fine antitumor efficacy was achieved with low side effect.

Abstract

Chelating Cu from tumors has been verified as an effective and promising strategy for cancer therapy through antiangiogenesis. However, systematic removal Cu by injecting with Cu chelators will result unavoidable side effects, since Cu is indispensable to the body. In this work, a micelle targeting to tumors' newborn vessels based on a polypeptide was developed to co-load DOX and Probe X, which can go through an “OFF-to-ON” procedure to report the Cu⁺-capture events in vivo in a real-time way by giving near infrared (NIR) fluorescence and photoacoustic signal. By co-delivering antiangiogenesis and chemotherapeutic reagents, the tumor can be significantly suppressed, meanwhile with a low systematic toxicity. Hopefully, this work can offer new insights in designing sophisticated antitumor strategy.

Graphical abstract

Introduction

Cu concentration, which is found extraordinarily elevated in many types of tumor, often correlates with tumor burden and recurrence [1,2]. Plentiful research suggests that Cu can promote the proliferation and migration of endothelial cells among the tumor sites [3], and Cu is also indispensable for tumor cells to secrete angiogenic factors [4]. There are already research on animal models focusing on suppressing tumor angiogenesis direct by inhibiting in vivo copper trafficking [5]. A famous case under clinical trial is tetrathiomolybdate, a strong copper chelator, which has been proved able to suppress angiogenesis and tumor growth via copper deficiency [6].

Chelating Cu can remarkably lower Cu concentration systematically by promoting the excretion after chelating [7]. However, Cu is a required element for life and an essential element functionalizing in most aerobic organisms, mainly playing as a structural and catalytic cofactor, and consequently involved in many biological pathways. Notably, lowering the Cu concentration systematically by simply injecting with Cu chelators was already found to cause severe side effects, including erythra, optic neuritis, emesis and leucopenia [8]. From another perspective, without enough reliable information on “when, where, how and how much” of the chelators' fuctionalization, clinical medication could be implicit [9]. As to our knowledge, there are very few reported systems concerning chelating Cu in vivo meanwhile with a reporting system [10]. It remains challenging to integrate a specific drug carrier to deliver Cu chelators to tumors to avoid systematic toxicity and meanwhile provide the real-time information on pharmacodynamics.

Cu, especially its monovalent form Cu⁺, serves as a critical signaling species in cells and can function in multiple organelles of tumor cells [11,12]. Cu⁺ plays a more important role than Cu²⁺ in tumor development and metastasis. On the other hand, in tumors' reducing microenvironment, Cu can stay in the Cu⁺ state [13] and be stabilized with GSH or Vitamin C within the cytosol [14]. Removal of Cu⁺ is more direct and Cu concentration will be thus synchronously lowered through the redox balance of Cu⁺-Cu²⁺ [15].

From both clinic and theory, the synergistic therapy of chemotherapy and antiangiogenesis is reasonable, where chemotherapy directly kills tumor cells and antiangiogenesis modulates the microenvironment by reducing new vessels that support tumors [16]. In clinic, there are already attempts trying to combine chemotherapy and antiangiogenesis together to suppress the cancer progress [17], while the clinical therapeutic efficacy by using combinations (such as bevacizumab/5-fluorouracil and trastazumab/paclitaxel for treating metastatic colorectal/breast cancers) has been well approved [18]. However, it is still challenging in specifically delivering chemotherapeutic agents and Cu chelators to the tumor sites for better synergy, since they often perform greatly different physical properties. The strategy can be further pushed forward by the targeting nanomedicine technology with finer efficacy and improved biocompatibility.

In this work, we co-loaded DOX and a Cu⁺ Probe X (both as the chelator and tracer) into a micellar system built from a body-friendly polypeptide amphiphile (Scheme 1 and Fig. 1). The micelle was integrated with cRGD to target tumors' neoangiogenesis, based on which, the chemotherapeutics and chelator can be delivered to tumor simultaneously. The Cu⁺ chelating events can be monitored by NIR fluorescence and photoacoustic imaging in a non-invasive and real-time way. After treatment, Cu level and oxygen saturation were significantly down-regulated in tumors, and the antineoangiogenesis mechanism was studied. The improved combined anti-tumor efficacy from antiangiogenesis and chemotherapy was achieved. Finally, the in vivo toxicity was also investigated. These findings, hopefully, can provide new insights to design, fabricate and tailor artificial antitumor nanosystems.

Section snippets

Materials

N₃-PEG-NH₂ (MW 5000) and PEG-NH₂ (MW 5000) were purchased from JenKem Technology Co. Ltd. (Beijing, China). Lys (cbz), phe, triphosgene and cyclohexanone were from Aladdin (Shanghai, China). HBr, AcOH, NaOH, NaOAc and CuI were from Energy Chemical (Shanghai, China). Thiourea, POCl₃, ethyl 2-hydroxyethyl sulfide, PMDETA and VcNa were commercially available from Sigma-Aldrich (Shanghai, China). Deuterium reagents for nuclear magnetic resonance (NMR) spectroscopy including CDCl₃ and DMSO-δ₆, CD₃OD

Design, theoretical calculation and synthesis

We first started the project by selecting a specific chelating ligand, as well as a tracker, for Cu⁺ both in vitro and in vivo. With the aim of facially capturing metal ions, various types of ligands have been developed and widely applied in MRI and biomedical sciences, to name a few, 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA), 1,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA) [24] and tetrathiomolybdate (TM) [25]. Most of the developed chelating ligands, however, have

Conclusion

Tumor growth and metastasis greatly rely on neoangiogenesis, where Cu⁺ plays an important role [43]. In this work, we proposed a micelle system that can co-delivery of Cu⁺ chelator and chemotherapeutics as a new strategy for tumor treatment strategy. Human body can be compared to a black box, and once drug was injected into the system, it is important to know the fate, though currently with much difficulty. A specific Probe X, both as the chelator and “Off-to-ON” type tracer for Cu⁺, was

Acknowledgements

This work was supported by National Natural Science Foundation of China (81872808) and Program of Shanghai Academic Research Leader (18XD1400500), Shanghai Municipal Science and Technology Major Project (2018SHZDZX01) and ZJLab, Fudan-SIMM Joint Research Fund (FU-SIMM20182006) and Scientific Research Program of Shanghai Health and Family Planning Commission (20184Y0149).

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Citation Excerpt :
They participate in the physiological regulation of TME and significantly influence the angiogenesis, tumorigenesis, metastasis, and many other behaviors during tumor progression [16]. For example, copper (Cu) can promote tumor cells to secrete angiogenic factors and improve the proliferation and migration of endothelial cells among the tumor sites [17],iron (Fe) can combine with diversified proteins, such as hemoglobin, to ensure the execution of downstream functions, in addition to acting as an indispensable mediator for numerous enzymes (such as mitochondrial enzymes, detoxification enzymes, etc.) to trigger necessary catalytic reactions in TME [18–19]; manganese (Mn), a cofactor of manganese superoxide dismutase (MnSOD or SOD2), can join in the antioxidant reactions reinforcing tumor progression [16]; aluminum (Al) derivatives as potential immunomodulatory adjuvants can be used to enhance antitumor immunity [20,21]. Except for the biological functions, the unique chemical properties of transitional metallic elements have caught much more attraction.
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Co-delivery of Cu(I) chelator and chemotherapeutics as a new strategy for tumor theranostic

Highlights

Abstract

Graphical abstract

Introduction

Section snippets

Materials

Design, theoretical calculation and synthesis

Conclusion

Acknowledgements

Cancer Treat. Rev.

Biomaterials

J. Control. Release

Tetrahedron Lett.

Am. J. Clin. Nutr.

Biomaterials

J. Control. Release

J. Control. Release

Cancer Treat. Rev.

J. Control. Release

J. Control. Release

Advances in copper complexes as anticancer agents

Chem. Rev.

Targeting copper in cancer therapy: copper that cancer

Metallomics

Role of copper in angiogenesis and its medicinal implications

Curr. Med. Chem.

Structural biology of copper trafficking

Chem. Rev.

Tetrathiomolybdate inhibits copper trafficking proteins through metal cluster formation

Science

Targeting copper to treat breast cancer

Science

Treatment of Wilson disease with ammonium tetrathiomolybdate: III. Initial therapy in a total of 55 neurologically affected patients and follow-up with zinc therapy

Arch. Neurol.

A targetable fluorescent sensor reveals that copper-deficient SCO1 and SCO2 patient cells prioritize mitochondrial copper homeostasis

J. Am. Chem. Soc.

Discerning the chemistry in individual organelles with small-molecule fluorescent probes

Angew. Chem. Int. Ed.

A mitochondria-targeted turn-on fluorescent probe based on a rhodol platform for the detection of copper(i)

Org. Biomol. Chem.

Rapid detection of copper in biological systems using click chemistry

Small