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.

抽象的： 4-Mmethylumbelliferone (4-MU) (M.W. 176)，也称为 hymechromone 或 7-羟基-4-甲基香豆素，是一种具有利胆和解痉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，在欧洲和亚洲，它也被用作膳食补充剂以改善肝脏健康。已知 4-MU 可以抑制透明质酸 (HA) 的生物合成，部分是通过与尿苷二磷酸 (UDP) 竞争作为 UDP-葡萄糖醛酸基转移酶 (UGT) 的底物。 HA 由 HA 合酶从其前体 UDP-葡萄糖醛酸 (UGA) 和 UDP-N-乙酰基-葡萄糖胺合成，这两种前体都是由 UGT 介导的 UDP 分别转移至葡萄糖醛酸和 N-乙酰基-葡萄糖胺而产生。通过抑制 UGT，4-MU 耗尽 UGA 胞质池，从而减少 HA 合成。 4-MU 还被证明可以降低 HA 合酶的表达水平，尽管这些酶的下调对 HA 合成的影响尚不完全清楚。 在过去的十年中，据报道 4-MU 在体外和体内对多种癌症发挥抗肿瘤和肿瘤预防活性，包括前列腺癌、肾癌、胰腺癌、卵巢癌、结肠癌、肝癌和皮肤癌。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 信号传导在肿瘤发生、生长、侵袭和新血管形成中发挥关键作用。在更晚期和更具侵袭性的癌症中，HA 和 HAS 水平更高。 Vinata Lokeshwar 博士及其同事之前使用转基因前列腺腺癌 (TRAMP) 小鼠模型报道，在第 8 周、12 周或22 周并持续到 28 周龄，以治疗时间依赖性方式显着减少了前列腺肿瘤发生，且没有明显的毒性迹象。从 8 周或 12 周到 28 周龄用 4-MU 治疗的 TRAMP 小鼠具有组织学正常的前列腺，具有低度前列腺上皮内瘤变 (PIN) 区域，这被认为是 8 周龄 TRAMP 小鼠前列腺的特征，没有任何器官转移，而载体对照小鼠被发现患有前列腺浸润性腺癌，并在肾、肝和肺中出现转移灶。尽管 4-MU 治疗在 28 周后停止，但 4-MU 的肿瘤抑制作用在 TRAMP 小鼠中持续观察到 52 周龄。此外，在 4-MU 治疗的小鼠中观察到的抗肿瘤功效与 HA 表达水平降低相关，伴随着前列腺组织中 Ki67 阳性细胞和微血管密度计数的减少以及 E-钙粘蛋白表达水平的增加。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用途。 4-MU 的药代动力学和安全性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进行过研究。临床证明，每日短期每日服用 1200 毫克至 2400 毫克 4MU 具有出色的安全性。对 4-MU 效用产生负面影响的因素是其半衰期短（约 30 分钟）和口服生物利用度低，据报道人体中的生物利用度低于 3%。 4-MU 通过 UGT 广泛代谢为葡萄糖醛酸 (4-MUG) 或硫酸盐形式 (4-MUS)。葡萄糖醛酸化被认为占 4-MU 代谢的 90% 以上。尽管 4-MUG 的药代动力学 (PK) 尚未得到充分研究，但 4-MUG 可通过胆汁消除，并可能从肠道重吸收（肠肝再循环），最终通过肾脏消除。考虑到 4-MU 抗肿瘤作用模式的工作假设需要非代谢形式的 4-MU 分子作为 UGT 的竞争性底物，如果 4-MU 的新制剂能够显着提高其肿瘤预防活性。 -MU可以开发用于靶向淋巴系统，同时避免肝脏中的首过代谢。这种淋巴靶向药物递送方法有望增加非肝器官中的 4-MU 浓度。 根据奥古斯塔大学佐治亚医学院的 Vinata Lokeshwar 博士向 NCI PREVENT Cancer 临床前药物开发计划提交的申请，开发了 4-MU、4-MU 肉豆蔻酸酯 (MUM) 的新配方通过与 PREVENT 协调的 DCP 存储库计划。 DCP Repository 进行的 MUM 初步 PK 研究表明，在多剂量小鼠研究中，4-MU 及其代谢物 MUG 在膀胱中积累较高，其次是在前列腺中。 数据表明 MUM 可能有助于预防浸润性膀胱癌。目前的研究重点是新配方的4-MU（MUM）的进一步开发，包括使用膀胱肿瘤发生的临床前模型评估其PK和药效（PD）曲线以及肿瘤预防功效。