Preclinical development of a chemopreventive agent, 4-methylumbelliferone(4-MU) HHSN2512015000361 TORFP: 2018-E07 PP: 9/17/18 to 9/16/19

化学预防剂 4-甲基伞形酮 (4-MU) 的临床前开发 HHSN2512015000361 TORFP: 2018-E07 PP: 9/17/18 至 9/16/19

• 批准号: 10006482
• 负责人: Clinton Grubbs
• 金额: $ 85.72万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-17 至 2021-09-16
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10006482
• 关键词: 4-methylumbelliferone; Age; Anabolism; Area; Asia; Biliary; Biological Availability; Bladder; Cells; Characteristics; Chemopreventive Agent; Cholecystectomy; Clinical; Clinical Trials; Colon; Consumption; Coumarins; Data; Development; Dose; Down-Regulation; Drug Delivery Systems; Drug Kinetics; Drug Targeting; Dyskinetic syndrome; E-Cadherin; Enzymes; Europe; Evaluation; Formulation; Glucosamine; Glucuronic Acids; Glucuronides; Glucuronosyltransferase; Goals; Growth; Half-Life; Health; Histologic; Human; Hyaluronic Acid; Intestines; Kidney; Liver; Lung; Lymphatic; Lymphatic System; Malignant Neoplasms; Malignant neoplasm of prostate; Malignant neoplasm of urinary bladder; Mediating; Metabolism; Mus; Myristates; Neoplasm Metastasis; Oral; Oral Administration; Organ; Ovary; Pancreas; Patients; Pharmacodynamics; Pharmacotherapy; Play; Pre-Clinical Model; Preclinical Drug Development; Prevention; Preventive; Prodrugs; Program Development; Property; Prostate; Prostate Adenocarcinoma; Prostatic Intraepithelial Neoplasias; Recycling; Reporting; Rodent Model; Role; Safety; Signal Transduction; Skin; Sodium; Sodium Chloride; Spasmolytics; Tissues; Toxic effect; Treatment Protocols; Universities; Unspecified or Sulfate Ion Sulfates; Uridine Diphosphate; Uridine Diphosphate Glucuronic Acid; Uridine Diphosphate N-Acetylglucosamine; biomarker evaluation; density; dietary supplements; efficacy evaluation; efficacy testing; healthy volunteer; improved; in vitro activity; in vivo; medical schools; mouse model; negative affect; neovascularization; optimal treatments; pharmacokinetics and pharmacodynamics; preclinical development; programs; prostate carcinogenesis; repository; transgenic adenocarcinoma of mouse prostate; tumor; tumor initiation; tumorigenesis

Abstract: 4-Methylumbelliferone (4-MU) (M.W. 176), also known as hymechromone or 7-hydroxy-4-methylcoumarin, is a coumarin derivative with choleretic and spasmolytic properties. It is an approved drug for the treatment of biliary dyskinesia in Europe and Asia, where it is also consumed as a dietary supplement to improve liver health. 4-MU is known to inhibit biosynthesis of hyaluronic acid (HA), in part by competing with uridine diphosphate (UDP) as a substrate for UDP-glucuronosyltransferase (UGT). HA is synthesized by HA synthases from its precursors, UDP-glucuronic acid (UGA) and UDP-N-acetyl-glucosamine, both of which are generated by the UGT-mediated transfer of UDP to glucuronic acid and N-acetyl-glucosamine, respectively. By inhibiting UGT, 4-MU depletes the cytosolic pool of UGA, thereby reducing HA synthesis. 4-MU has also been shown to reduce the expression levels of HA synthases, although the impact of downregulations of these enzymes on HA synthesis is not fully understood. For over the last decade, 4-MU has been reported to exert anti-tumor and tumor-preventive activities in vitro and in vivo against wide spectrum of cancers, including cancers of prostate, kidney, pancreas, ovary, colon, liver and skin. These antitumor activities have been attributed to the ability of 4-MU to inhibit HA biosynthesis, as HA signaling is known to play a critical role in tumor initiation, growth, invasion, and neovascularization. Higher levels of HA and HAS have been demonstrated in more advanced and aggressive cancers. Using the transgenic adenocarcinoma of the prostate (TRAMP) mouse model, Dr. Vinata Lokeshwar and colleagues have previously reported that daily oral administration of 4-MU (450 mg/kg/day by gavage), started at 8 weeks, 12 weeks, or 22 weeks and continued through 28 weeks of age, significantly reduced prostate tumorigenesis without signs of overt toxicity in a treatment-timing dependent manner. TRAMP mice treated with 4-MU from 8 weeks or 12 weeks through 28 weeks of age had histologically normal prostates with areas of low-grade prostatic intraepithelial neoplasia (PIN), which is considered characteristic of the 8-week old TRAMP mice prostate, without any organ metastasis, while the vehicle control mice were found to have invasive adenocarcinoma of the prostate with metastatic lesions in the kidney, liver, and lung. The tumor suppressive effect of 4-MU continued to be observed in the TRAMP mice through 52 weeks of age even though the 4-MU treatment was stopped after 28 weeks. Moreover, the antitumor efficacy observed in the 4-MU treated mice was associated with the decreased levels of HA expression, accompanied by reduced Ki67-positive cells and microvessel density counts with the increased level of E-cadherin expression in the prostate tissue. These findings supported the potential utility of 4-MU as a nontoxic oral chemopreventive agent for prostate and other cancers. Pharmacokinetics and safety of 4-MU have previously been studied in clinical trials involving healthy volunteers and patients requiring cholecystectomy. Excellent safety profiles have been clinically demonstrated for short-term daily administrations of 4MU at 1200 mg to 2400 mg per day. Factors negatively affecting the utility of 4-MU are its short half-life (~30 min) and low oral bioavailability, which has been reported to be less than 3% in humans. 4-MU is extensively metabolized to a glucuronic acid (4-MUG) via UGT or a sulfate form (4-MUS). Glucuronidation is believed to account for more than 90% of 4-MU metabolism. 4-MUG is biliary eliminated and likely undergoes reabsorption from the intestine (enterohepatic recycling) for eventual elimination by the kidney, although pharmacokinetics (PK) of 4-MUG has not been well studied. Considering the working hypothesis for the mode of antitumor action of 4-MU, which requires a non-metabolized form of 4-MU molecule to serve as a competitive substrate for UGT, its tumor preventive activity may be significantly improved if a new formulation of 4-MU can be developed to target the lymphatic system while avoiding the first pass metabolism in the liver. Such a lymphatic targeting drug delivery approach is expected to increase 4-MU concentrations in non-hepatic organs. As a result of an application to the NCI PREVENT Cancer Preclinical Drug Development Program submitted by Dr. Vinata Lokeshwar of the Medical College of Georgia, Augusta University, a new formulation of 4-MU, 4-MU myristate (MUM), has been developed through the DCP Repository Program in coordination with PREVENT. The preliminary PK study of MUM conducted by the DCP Repository demonstrated that higher accumulations of 4-MU and its metabolite MUG were demonstrated in the bladder followed by the prostate in a multi-dosing mouse study. The data suggested that MUM may be useful for the prevention of invasive bladder cancer. The current study is focused on the further development of the newly formulated 4-MU, MUM, including the evaluation of its PK and pharmacodynamics (PD) profiles and tumor preventive efficacy using a preclinical model of bladder tumorigenesis.

摘要： 4-甲基伞形酮（4-MU）（M.W. 176），也称为羟甲香豆素或7-羟基-4-甲基香豆素，是一种具有利胆和痉挛特性的香豆素衍生物。它是欧洲和亚洲批准用于治疗胆道运动障碍的药物，在那里它也被用作膳食补充剂以改善肝脏健康。已知4-MU抑制透明质酸（HA）的生物合成，部分通过与尿苷二磷酸（UDP）竞争作为UDP-葡糖醛酸基转移酶（UGT）的底物。HA由HA脱氢酶从其前体UDP-葡萄糖醛酸（UGA）和UDP-N-乙酰基-葡糖胺合成，这两种前体均通过UGT介导的UDP分别转移至葡萄糖醛酸和N-乙酰基-葡糖胺而产生。通过抑制UGT，4-MU耗尽UGA的胞质池，从而减少HA合成。4-MU还显示降低HA酶的表达水平，尽管这些酶的下调对HA合成的影响尚未完全了解。 在过去的十年中，已经报道了4-MU在体外和体内对广泛的癌症（包括前列腺癌、肾癌、胰腺癌、卵巢癌、结肠癌、肝癌和皮肤癌）发挥抗肿瘤和肿瘤预防活性。这些抗肿瘤活性归因于4-MU抑制HA生物合成的能力，因为已知HA信号传导在肿瘤起始、生长、侵袭和新血管形成中起关键作用。在更晚期和侵袭性的癌症中已经证明了更高水平的HA和HAS。 使用转基因前列腺腺癌（TRAMP）小鼠模型，Vinata Lokeshwar博士及其同事先前报告称，从8周龄、12周龄或22周龄开始每天经口给予4-MU（450 mg/kg/天，灌胃）并持续至28周龄，可显著降低前列腺肿瘤发生，且无明显毒性迹象，且具有治疗时间依赖性。用4-MU处理的8周龄或12周龄至28周龄的TRAMP小鼠具有组织学上正常的前列腺，具有低度前列腺上皮内瘤形成（PIN）区域，其被认为是8周龄TRAMP小鼠前列腺的特征，没有任何器官转移，而发现载体对照小鼠具有前列腺的侵袭性腺癌，在肾、肝、和肺在TRAMP小鼠中持续观察到4-MU的肿瘤抑制作用，直至52周龄，即使在28周后停止4-MU治疗。此外，在4-MU处理的小鼠中观察到的抗肿瘤功效与HA表达水平降低相关，伴随着Ki 67阳性细胞和微血管密度计数减少以及前列腺组织中E-钙粘蛋白表达水平增加。这些发现支持4-MU作为前列腺癌和其他癌症的无毒口服化学预防剂的潜在效用。 先前已在涉及健康志愿者和需要胆囊切除术的患者的临床试验中研究了4-MU的药代动力学和安全性。临床上已证明4 MU每日1200 mg至2400 mg短期给药具有良好的安全性特征。负面影响4-MU效用的因素是其半衰期短（约30分钟）和口服生物利用度低，据报道在人体中低于3%。4-MU通过UGT或硫酸盐形式（4-MUS）广泛代谢为葡萄糖醛酸（4-MUG）。据信葡萄糖醛酸化占4-MU代谢的90%以上。4-MUG通过胆汁消除，并可能经历肠重吸收（肝肠再循环），最终通过肾脏消除，尽管4-MUG的药代动力学（PK）尚未得到充分研究。考虑到4-MU的抗肿瘤作用模式的工作假设，其需要4-MU分子的非代谢形式作为UGT的竞争性底物，如果能够开发出靶向淋巴系统的4-MU新制剂，同时避免肝脏中的首过代谢，则其肿瘤预防活性可能会显著改善。预期这种淋巴靶向药物递送方法增加非肝器官中的4-MU浓度。 由于奥古斯塔大学格鲁吉亚医学院的Vinata Lokeshwar博士向NCI PREVENT癌症临床前药物开发计划提交的申请，通过DCP储存库计划与PREVENT协调开发了4-MU，4-MU肉豆蔻酸酯（MUM）的新制剂。DCP Repository进行的MUM初步PK研究表明，在多次给药小鼠研究中，4-MU及其代谢物MUG在膀胱中的蓄积较高，其次是前列腺。 这些数据表明，MUM可能有助于预防浸润性膀胱癌。目前的研究重点是进一步开发新配制的4-MU，MUM，包括使用膀胱肿瘤发生的临床前模型评价其PK和药效学（PD）特征以及肿瘤预防疗效。