CN-120241652-B - Protopanaxatriol nano delivery system targeting Twist1-Ku70-Sirt1 complex and application thereof in colorectal cancer treatment related to colonitis

Abstract

The invention relates to the field of medicines, in particular to a protopanaxatriol nano delivery system of a targeted Twist1-Ku70-Sirt1 compound and application of the nano delivery system in treating colonitis related colorectal cancer, wherein the nano delivery system comprises (a) protopanaxatriol nanocrystals, (b) lipoic acid modified chitosan and (c) a cross-linking agent, wherein the average particle size of the protopanaxatriol nanocrystals is 100-200 nm, the modification degree of the lipoic acid modified chitosan is 15-25, the cross-linking agent is sodium tripolyphosphate, and the average particle size of the nano delivery system is 300-500 nm.

Inventors

• XUN QINGQING
• LIU YANRONG
• XU SHAOSHAO
• LI YAFANG
• YIN DAN
• Miao Chengzhen
• DU LINLIN

Assignees

• 济宁医学院附属医院

Dates

Publication Date

20260508

Application Date

20250421

Claims (4)

1. 1. A protopanaxatriol nanodelivery system targeting a Twist1-Ku70-Sirt1 complex, the nanodelivery system comprising: (a) A protopanaxatriol nanocrystal prepared by an ultrasonic-assisted anti-solvent precipitation method, wherein ethanol is used as a solvent, ultrapure water is used as an anti-solvent, and polyvinylpyrrolidone K30 or polysorbate 80 is used as a stabilizer; (b) Lipoic acid modified chitosan with molecular weight of 100-200 kDa and deacetylation degree not less than 90%, synthesized through EDC/NHS activated coupling process, wherein EDC is 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide, NHS is N-hydroxysuccinimide, and (C) A cross-linking agent which is sodium tripolyphosphate; The average particle size of the protopanaxatriol nanocrystals is 100-200 nm, the modification degree of the lipoic acid modified chitosan is 15-25%, the average particle size of the nano delivery system is 300-500 nm, the mass ratio of the protopanaxatriol nanocrystals to the lipoic acid modified chitosan is 1:2-1:5, the mass ratio of the lipoic acid modified chitosan to sodium tripolyphosphate is 3:1-5:1, the Zeta potential absolute value of the nano delivery system is more than or equal to 30 mV, the polydispersity coefficient is less than 0.3, the protopanaxatriol loading capacity is more than or equal to 10%, and the encapsulation rate is more than or equal to 80%; The protopanaxatriol in the nano-delivery system binds to tyrosine 320 of Ku70 and glutamic acid 32 of Sirt1, thereby inhibiting the formation of a Twist1-Ku70-Sirt1 ternary protein complex.
2. 2. The nano-delivery system of claim 1, wherein the nano-delivery system releases <20% of protopanaxatriol within 4 hours in simulated gastric fluid pH 1.2 and in intestinal fluid pH 6.8 and releases >80% of protopanaxatriol within 24 hours in simulated intestinal fluid pH 7.4 with the addition of colonic bacterial enzymes, exhibiting colonic targeted release characteristics.
3. 3. A method of preparing the nano-delivery system of claim 1 or 2, comprising the steps of: (a) Dissolving protopanaxatriol in ethanol, adding a stabilizer, and adding the protopanaxatriol solution into ultrapure water under ultrasonic conditions to prepare protopanaxatriol nanocrystals; (b) Activating lipoic acid through EDC/NHS, and reacting the activated lipoic acid with chitosan to prepare lipoic acid modified chitosan; (c) Dispersing the raw panaxatriol nanocrystals prepared in step (a) in a solution containing lipoic acid modified chitosan prepared in step (b), then adding sodium tripolyphosphate solution for crosslinking to form a nano-delivery system, and (D) Collecting and purifying the nano-delivery system.
4. 4. Use of a nanodelivery system according to claim 1 or 2 in the manufacture of a medicament for the treatment of colorectal cancer associated with colitis, wherein the nanodelivery system works by binding protopanaxatriol to tyrosine 320 of Ku70 and glutamic acid 32 of Sirt1, targeted inhibition of the formation of a Twist1-Ku70-Sirt1 ternary protein complex, reduction of stem cell characteristics and tumorigenesis of cancer cells, wherein the medicament is an oral formulation, administered at a dose equivalent to 10-20 mg/kg/day of protopanaxatriol for the treatment of colorectal cancer associated with colitis induced by AOM-DSS.

Description

Protopanaxatriol nano delivery system targeting Twist1-Ku70-Sirt1 complex and application thereof in colorectal cancer treatment related to colonitis Technical Field The invention relates to the field of medicines, in particular to a protopanaxatriol nano delivery system targeting a Twist1-Ku70-Sirt1 complex and application of the nano delivery system in treating colonitis related colorectal cancer. Background Colitis-related colorectal cancer (Colitis-associated colorectal cancer, CAC) is one of the most serious complications in patients with Inflammatory Bowel Disease (IBD), and chronic inflammation in the long term is a major risk factor for CAC. Epidemiological data show that inflammatory bowel disease patients are at significantly higher risk of developing colorectal cancer than the general population, with lifelong colorectal cancer incidence of ulcerative colitis patients being as high as 18%. Compared with sporadic colorectal cancer, CAC has the characteristics of early onset, multiple onset, poor prognosis and the like, and brings great burden to patients. Existing treatment regimens for CAC mainly include anti-inflammatory treatment, chemotherapy, surgical resection, etc., however these treatments have limited efficacy and are often accompanied by serious adverse reactions. Currently, there is a clinical lack of targeted therapeutic drugs against CAC-specific pathological mechanisms, especially drugs capable of inhibiting tumor stem cell characteristics and tumorigenesis, which is a major challenge faced by CAC treatment. In recent years, various transcription factors and signal paths in tumor microenvironment are found to play an important role in CAC development. Among them, twist1 is a key regulator of epithelial-mesenchymal transition (EMT), is highly expressed in various cancers and is associated with disease progression and poor prognosis. Studies have shown that Twist1 promotes maintenance of stem cell characteristics, invasion metastasis and resistance development in tumor cells in colorectal cancer. Liu et al (Oncotarget, 2017) reported that Twist1 promotes multi-drug resistance in colorectal cancer by upregulating ATP-binding cassette transporters (such as ABCB1 and ABCC 1). Chen et al (SCIENTIFIC REPORTS, 2014) found that Sirt1 was highly expressed in colorectal cancer tissue and was significantly correlated with patient prognosis and that Sirt1 was co-localized to colorectal cancer stem cell marker CD 133. On the molecular mechanism level, roth et al (Oncotarget, 2016) reported for the first time that SIRT1 and LSD1 competitively regulate Ku70 function, affecting DNA repair and tumor cells to acquire the molecular mechanism of drug-resistant mutation. Studies have shown that Ku70 is a key protein of the non-homologous end joining (NHEJ) DNA repair pathway, while SIRT1 enhances its DNA repair function by deacetylating Ku 70. In addition, cohen et al (Science, 2004) studies demonstrated that SIRT1 regulates the sensitivity of cells to apoptotic stimuli by deacetylating Ku 70. However, these studies have focused mainly on the binary protein interaction level, and there is no in-depth study on potential ternary protein complexes and their role in tumorigenesis. The effective components of the traditional Chinese medicine have long history and broad prospect in tumor treatment. Protopanaxatriol (20S-protopanaxatriol, PPT) is one of the major active metabolites of ginsenoside, and has been demonstrated to have a variety of pharmacological activities including anti-tumor, anti-inflammatory, anti-oxidant, etc. The studies of Zhao et al (Molecules, 2010) show that PPT and its derivatives have significant proliferation inhibition effects on various tumor cell lines. King et al (Journal of Experimental & CLINICAL CANCER RESEARCH, 2019) reported that 20 (S) -protopanaxatriol (g-PPT) in combination with EGFR-TKI could overcome EGFR-TKI resistance by reducing SCD 1-induced lipid accumulation. However, PPT, as a natural product, has disadvantages of poor water solubility, low bioavailability, short in vivo half-life, etc., which limits its clinical application. As a novel drug delivery platform, nano-delivery systems have made remarkable progress in recent years in improving bioavailability, enhancing targeting and controlling release of poorly soluble drugs. Gao Jun et al (journal of international pharmaceutical research, 2018) reviewed the progress of the use of nano-delivery systems in colorectal cancer treatment, indicating that nano-delivery systems can significantly increase drug enrichment at tumor sites and reduce systemic toxicity. Chitosan, a natural polysaccharide with good biocompatibility and biodegradability, has been widely used for the construction of drug delivery systems. Liu et al (International Journal of Nanomedicine, 2016) reported that chitosan nanoparticles could effectively enhance drug release at colon sites and improve therapeutic effects on colonic inflammation. However, t