Therapeutic Nanomedicine based Strategies for the Treatment of Atherosclerosis and Cancer
著者 SINDHU C PILLAI
学位授与大学 東洋大学
取得学位 博士
学位の分野 バイオ・ナノサイエンス融合
報告番号 32663甲第473号 学位授与年月日 2020‑09‑25
URL http://id.nii.ac.jp/1060/00012182/
Creative Commons : 表示 ‑ 非営利 ‑ 改変禁止 http://creativecommons.org/licenses/by‑nc‑nd/3.0/deed.ja
氏 名 （ 本 籍 地 ） SINDHU C PILLAI（インド）
学 位 の 種 類 博士（バイオ・ナノサイエンス融合）
報 告 ・ 学 位 記 番 号 甲第473号（甲（バ）第十一号）
学 位 記 授 与 の 日 付 2020年9月25日
学 位 記 授 与 の 要 件 本学学位規程第3条第1項該当
学 位 論 文 題 目
Therapeutic Nanomedicine based Strategies for the Treatment of Atherosclerosis and Cancer（和訳：アテローム性動脈硬化およ びがん治療のための戦略的ナノ治療薬）
論 文 審 査 委 員 主査 教授 Ph.D. D. Sakthi Kumar 副査 教授 工学博士 前川 透
副査 特任准教授 博士（ﾊﾞｲｵ･ﾅﾉ
ｻｲｴﾝｽ融合） Sheikh Mohamed Mohamed 副査 教授 博士（工学） 花尻 達郎
副査 客員教授 博士（工学） 水木 徹 副査 教授 博士（工学） 森本 久雄
Graduate School of Interdisciplinary New Science
学位論文審査結果報告書〔甲〕
Examiners’ Reports on Doctoral Thesis
【論文審査】 Summary and Review of the thesis
Ms. Sindhu C Pillai’s thesis titled “Therapeutic Nanomedicine based Strategies for the Treatment of Atherosclerosis and Cancer” present about the nanomedicine developed based on natural products mainly Curcumin, Resveratrol and BioPerine. She used the nanoformulations developed by the natural products to find out the solution for atherosclerosis and cancer.
As per several studies conducted worldwide, cancer and cardiovascular diseases are primarily found to be responsible for nearly two thirds of all deaths. The estimated costs involving the treatment of these two diseases have been predicted to hit almost 2.8 million dollars in the global economy by 2020. It is revealed from the detailed studies that inflammation or the activation of inflammatory pathway are the major contributors to the initiation and development of these diseases arise leading to death. Unfortunately, the current therapies employed to curb atherosclerosis and cancer are not found to be fully effective. Moreover, these are also found to be associated with other issues that include the adverse side effects and toxicity. By using the nutraceuticals we can solve this problem greatly, however, due to its low bioavailability and poor solubility it cannot be employed for the job as such. With the help of nanotechnology and modification of the surface of these materials we can improve the solubility and bioavailability and by using the selective targeting we can deliver the drug to the targeted sites. In this thesis Ms. Sindhu used the nutraceuticals such as curcumin with a natural bio enhancer BioPerine that improves bioavailability and absorption resulting in enhanced efficacy.
She used BioPerine based-hybrid polymeric nanoparticles (NPs) for reversing multi-drug resistivity in breast cancer and compared and validated its effect with conventional drug such as verapamil hydrochloride.
Ms. Sindhu divided the thesis into 6 chapters.
Chapter 1 Ms. Sindhu presented a thorough review study in relation to her work and presented the importance of her work. In the introduction chapter she mentioned about the chronic inflammatory diseases like atherosclerosis and cancer and discussed about the onset of the same. These menace actually taking the lives of human beings due to the changes in lifestyle including diet that can cause the lipid accumulation leading to the development of atherosclerosis while, mutations within the body causes cancer. The inflammation and a series of shared molecular pathways of both diseases lead to a new breakthrough towards the treatment. Pro inflammatory cytokines, chemokines, chemoattractant-proteins and tumor associated macrophages (TAMs) have been found to induce chronic inflammation and also serve as the key component of the tumor microenvironment in atherosclerosis and cancer respectively. Although currently there are many treatments
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evolved for both the diseases, they are still uncontrolled. Thus, an alternative solution to control these chronic inflammatory diseases is a critical necessity. Nutraceuticals from natural compounds have been found as a promising agent in the field of pharmaceutical therapy to combat these deadly diseases.
In Chapter 2 Ms. Sindhu mentioned about the principle concept and Instrumentation techniques that have been adopted by her to complete her research work. She used techniques such as SEM, TEM, XPS, XRD, FTIR and to characterize the nanoparticles. She carried out invitro cell culture studies in cancer cell lines and monocytes that comprises of cytotoxicity assay, cellular uptake, optical microscopy and others to demonstrate the effectiveness of the nano drugs developed by her.
In Chapter 3 she mentioned about the work that she conducted for the co-delivery of Curcumin and BioPerine by using PLGA nanoparticles that prevents atherosclerotic foam cell formation by reducing CD-36 receptor expression and NF-kB Signaling in Human Macrophages. Atherosclerosis has been characterized by the development of cholesterol-rich arterial plaques which is a major cause of heart disease. Inflammatory cytokines are highly upregulated by the arterial plaques that are secreted by the macrophages that finally become cholesterol-laden foam cells. Current therapeutics aimed at treating atherosclerosis resulted in side effects such as organ damage, diabetes, and often fail to fully reverse or repair the damaged arteries. Alternative options such as the use of nutraceuticals could offer some edge over the current synthetic substances in preventing the above-mentioned associated side effects. Curcumin, a potent polyphenol present in Curcuma longa demonstrated remarkable anti-atherosclerotic activity via anti-inflammatory and anti-oxidative properties. The bioavailability and poor water solubility of curcumin limit its clinical translational purposes, which can circumvent effectively by nano-drug delivery systems that can target atherosclerotic plaque sites.
To show the effectiveness of the nutraceuticals, she chose curcumin and to increase its bioavailability included BioPerine (derived from Piper nigrum) which is a natural bio enhancer inside a polymeric nano-drug delivery system for targeting atherosclerotic plaque sites. She selected two different ratios of curcumin: BioPerine to study its comparative effect on the inhibition of oxidized low-density lipoprotein (Ox-LDL) induced foam cell formation and studied the therapeutic efficacy in THP-1 monocyte-derived macrophages via different in vitro cell studies. Her studies demonstrated that the administration of BioPerine alongside curcumin via PLGA nanoparticles (NPs) imparts a reduction in macrophage-mediated foam cell formation, relative cholesterol content and the inflammatory pathways when administered as preventive medicine. This result clearly showing the importance of natural-based compounds towards the therapeutic intervention against the atherosclerotic activity.
In Chapter 4 Ms. Sindhu describing about the anti-cancerous activity of Resveratrol-BioPerine loaded PEG coated Polymeric Nanoparticles in Pancreatic and Breast Cancer. The enhanced efficacy of BioPerine has been
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successfully proven in cardiovascular disease. Ms. Sindhu modified the nanoparticle to check the efficiency of bioperine even in cancer by encapsulated resveratrol a natural drug into PLGA PEG NPs. Resveratrol (RES) is a polyphenolic compound found in grapes, that has potent anti-oxidant, anti-cancerous and anti- inflammatory activities. During this study, she analyzed the activity of resveratrol-bioperine PLGA PEG NPs towards breast cancer cell lines MDA-MB 353, MCF-7 and pancreatic cell lines. The PLGA RES Bioperine PEG NPs were characterized for their size, shape and chemical interaction by SEM, TEM, DLS, FTIR, and XRD analysis respectively. PLGA RES-BioPerine -PEG NPs arrested cells in G2/M phase and blocks the microtubule polymerization invitro. It reduces the ROS activity in turns reduces inflammation, reduce the cell proliferation and autophagy. From the experimental results she inferred that PLGA RES-Bioperine -PEG NPs increased the efficacy towards cancer cells as a novel anti-cancer therapy.
In the Chapter 5 Ms. Sindhu is mentioning about the BioPerine encapsulated nanoformulation for overcoming drug-resistant Breast Cancers. The drug resistance due to tumor heterogeneity often creates impediments to traditional therapies making it a challenging issue for cancer cure. Breast cancer often faces the challenges of current therapeutic interventions owing to its multiple complexities and high drug resistivity for example against drugs like trastuzumab and tamoxifen. Drug resistance in the majority of breast cancers is often aided by the overtly expressed P-glycoprotein (P-gp) that guides in the rapid drug efflux of chemotherapy drugs.
Despite continuous endeavors and ground-breaking achievements in the pursuit of finding better cancer therapeutic avenues, drug resistance is still a menace to hold back. Among the newer therapeutic approaches, the application of phytonutrients such as alkaloids to suppress P-gp activity in drug-resistant cancers has found an exciting niche in the arena of alternative cancer therapies. To suppress the P-gp activity, Ms. Sindhu developed a nanoformulation having black pepper alkaloid derivative known as BioPerine-loaded chitosan (CS)-polyethylene glycol (PEG) coated polylactic acid (PLA) hybrid polymeric nanoparticle to improve the bioavailability of BioPerine and its therapeutic efficacy in suppressing P-gp expression in MDA-MB 453 breast cancer cell line. The developed CS-PEG-BioPerine-PLA nanoparticles showed a smooth spherical morphology with an average size of 316 nm, with improved aqueous solubility, and provided sustained BioPerine release. The CS-PEG-BioPerine-PLA nanoparticles also reported enhanced in vitro cytotoxicity and downregulation of P-gp expression in MDA-MB 453 cells compared to the commercial inhibitor verapamil hydrochloride. This clearly shows that BioPerine based nano-drug delivery system in combination with traditional therapies as a crucial approach to tackling multi-drug resistance in cancers.
In Chapter 6 Ms. Sindhu concluded and summarized all her work that she conducted during her doctoral course that presented in the thesis.
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【審査結果】 Examination Results
The thesis, submitted by Ms. Sindhu C Pillai titled “Therapeutic Nanomedicine based Strategies for the Treatment of Atherosclerosis and Cancer” describes about, the use of Curcumin, Resveratrol and Bioperine as the drug in the nanoformulation to work against atherosclerosis, to kill different cancer cells and to reduce the P-gp efflux pump actions to curb the drug resistance.
The results shown in the thesis are outstanding and the significant points in the present study are summarised below;
1. Developed nanoformulation of Curcumin and BioPerine by using PLGA nanoparticles that prevents atherosclerotic foam cell formation by reducing CD-36 receptor expression and NF-kB Signaling in Human Macrophages. This nanoformulation showed highly effective against atherosclerosis.
2. Developed Resveratrol-Bioperine loaded PEG coated Polymeric Nanoparticles against Pancreatic and Breast Cancer. This nanoformulation showed effectiveness in the cancer killing.
3. Developed BioPerine encapsulated nanoformulation for overcoming drug-resistant Breast Cancers by inhibiting P-gp efflux pump. This drug proved to be highly effective in comparison with commercial P-gp inhibitor verapamil hydrochloride.
4. Combination of Curcumin and Bioperine nanoformulation could be a potential candidate to prevent the atherosclerosis and the finding of P-gp inhibiting Bioperine nanoformulation indeed a great step in the field of nanodrug delivery to overcome the drug resistance.
Ms. Sindhu has already published 1 first author paper and 1 second author paper in internationally reputed journals. 1 more paper communicated based on her work which is under review. She will be submitting one more paper which is review paper soon. The results obtained in this study have been presented at 2 international conferences and symposia. Based on her work, she already received Inoue Enryo funding award from the university to conduct her work, which clearly shows the appreciation of the quality of her work.
Judging by the results shown in the thesis, the number of international papers published so far and the number of presentations at international conferences and symposia, the level of the research results is definitely high by international standards and the present results may well make a great contribution to the development of new drug formulations and its application in the atherosclerosis and cancer field. In conclusion, the thesis is considered to be a high quality, high standard one by international standards.
