Olaparib (AZD2281; KU0059436)   中文名称: 奥拉帕尼

PARP-2 ( IC50 = 1 nM ); PARP-1 ( IC50 = 5 nM ); tankyrase-1 ( IC50 = 1.5 μM ); Autophagy; Mitophagy

体外活性：奥拉帕尼可对抗 BRCA1 或 BRCA2 突变。奥拉帕尼对坦科聚合酶 1 不敏感 (IC50 >1 μM)。浓度为 30-100 nM 的奥拉帕尼可以消除 SW620 细胞中的 PARP-1 活性。与 BRCA1 和 BRCA2 丰富的细胞系（Hs578T、MDA-MB-231 和 T47D）相比，奥拉帕尼对 BRCA1 缺陷细胞系（MDA-MB-463 和 HCC1937）高度敏感。由于PARP抑制作用抑制了碱基切除修复，奥拉帕尼对KB2P细胞高度敏感，这可能导致DNA复制过程中单链断裂转化为双链断裂，从而激活BRCA2依赖性重组途径。激酶测定：向第 1 至第 10 列中添加 1 μL 奥拉帕尼（在 DMSO 中），并且仅将 1 μL DMSO 添加至阳性 (POS) 和阴性 (NEG) 对照孔（分别为第 11 列和第 12 列）预处理的 FlashPlate。 PARP-1 在缓冲液中按 1:40 稀释（缓冲液 B：10% 甘油 (v/v)、25 mM HEPES、12.5 mM MgCl2、50 mM KCl、1 mM DTT、0.01% NP-40 (v/v)、 pH 7.6）和 40 μL 添加到所有 96 个孔中（测定中的最终 PARP-1 浓度约为 1 ng/μL）。将板密封并在室温下摇动 15 分钟。随后，将 10 μL 阳性反应混合物（每孔 0.2 ng/μL 双链寡核苷酸 [M3/M4] DNA、5 μM NAD+ 最终测定浓度和每孔 0.075 μCi 3H-NAD+）添加到适当的反应孔中。孔（第 1-11 列）。将不含 DNA 寡核苷酸的阴性反应混合物添加到第 12 列（使用平均阴性对照值作为背景）。将板重新密封并在室温下再摇动 60 分钟以使反应继续进行。然后，向每孔中加入 50 μL 冰冷的乙酸 (30%) 以终止反应，并将板密封并在室温下再摇动 60 分钟。然后使用 TopCount 读板器以每分钟计数 (CPM) 确定与 FlashPlate 结合的氚化信号。细胞测定：奥拉帕尼的细胞毒性通过克隆形成测定来测量。使用前将奥拉帕尼溶解在 DMSO 中并用培养基稀释。将细胞（乳腺癌细胞系，包括 SW620 结肠细胞、A2780 卵巢细胞、HCC1937、Hs578T、MDA-MB-231、MDA-MB-436 和 T47D）接种在六孔板中并贴壁过夜。然后添加不同浓度的奥拉帕尼并将细胞孵育7-14天。之后对存活的菌落进行计数以计算IC50。

奥拉帕尼（10 mg/kg，口服）与替莫唑胺联合使用可显着抑制 SW620 异种移植物中的肿瘤生长。奥拉帕尼对 Brca1-/-;p53-/- 乳腺肿瘤显示出良好的反应（每天 50 mg/kg ip），而对 HR 缺陷的 Ecad-/-;p53-/- 乳腺肿瘤没有反应。奥拉帕尼甚至在荷瘤小鼠中没有表现出剂量限制性毒性。奥拉帕尼已用于治疗 BRCA 突变肿瘤，例如卵巢癌、乳腺癌和前列腺癌。此外，Olaparib 对 ATM（共济失调毛细血管扩张突变）缺陷型肿瘤细胞表现出选择性抑制作用，这表明它是治疗 ATM 突变淋巴肿瘤的潜在药物。

该测定评估了奥拉帕尼抑制 PARP-1 酶活性的能力。测量 PARP-2 活性抑制的另一种方法涉及使用 PARP-2 特异性抗体在白壁 96 孔板中结合重组 PARP-2 蛋白。添加 ³H-NAD⁺ DNA 后测量 PARP-2 活性。洗涤后添加闪烁剂以量化 ³H 掺入的核糖基化。创建了针对 tankyrase-1 的 AlphaScreen 测定，包括在 384 孔 ProxiPlate 测定中与生物素化 NAD⁺ 一起孵育 HIS 标记的重组 TANK-1 蛋白。通过添加 α 珠子结合 HIS 和生物素标签来产生邻近信号；该信号的丢失与 TANK-1 活性抑制直接相关。每个实验至少进行三次重复。

增强因子或 PF50 值是通过将对照生长中使用的烷化剂甲磺酸甲酯 (MMS) 的 IC50 除以 MMS 加 PARP 抑制剂的 IC50 来确定的。使用 HeLa B 细胞以固定 200 nM 浓度对 Olaparib 进行 MMS 筛选测试。在结直肠细胞系 SW620 上测试的奥拉帕尼浓度为 1、3、10、100 和 300 nM。磺胺罗丹明 B (SRB) 测定用于测量细胞生长。

Absorption, Distribution and Excretion
Following oral administration, olaparib is rapidly absorbed. After administration of a single 300 mg dose of olaparib, the mean (CV%) Cmax was 5.4 μg/mL (32%) and AUC was 39.2 μg x h/mL (44%). The steady state Cmax and AUC following a dose of 300 mg twice daily was 7.6 μg/mL (35%) and 49.2 μg x h/mL (44%), respectively. Tmax is 1.5 hours. A high-fat and high-calorie meal may delay Tmax, but does not significantly alter the extent of olaparib absorption.
Following a single dose of radiolabeled olaparib, 86% of the dosed radioactivity was recovered within a seven-day collection period, mostly in the form of metabolites. About 44% of the drug was excreted via the urine and 42% of the dose was excreted via the feces. Following an oral dose of radiolabeled olaparib to female patients, the unchanged drug accounted for 15% and 6% of the radioactivity in urine and feces, respectively.
The mean (± standard deviation) apparent volume of distribution of olaparib is 158 ± 136 L following a single 300 mg dose.
Following a single oral dose in patients with cancer, the mean apparent plasma clearance was 4.55 L/h.

---

Metabolism / Metabolites
Olaparib is metabolized by cytochrome P450 (CYP) 3A4/5 _in vitro_. Following an oral dose of radiolabeled olaparib to female patients, unchanged olaparib accounted for 70% of the circulating radioactivity in plasma. Olaparib undergoes oxidation reactions as well as subsequent glucuronide or sulfate conjugation. In humans, olaparib can also undergo hydrolysis, hydroxylation, and dehydrogenation. While up to 37 metabolites of olaparib were detected in plasma, urine, and feces, the majority of metabolites represent less than 1% of the total administered dose and they have not been fully characterized. The major circulating metabolites are a ring-opened piperazin-3-ol moiety and two mono-oxygenated metabolites. The pharmacodynamic activity of the metabolites is unknown.

---

Biological Half-Life
Following a single oral dose in patients with cancer, the mean terminal half-life was 6.10 hours.

Hepatotoxicity
In large clinical trials of olaparib, abnormalities in routine liver tests were uncommon with serum aminotransferase elevations occurring in 4% of patients and values above 5 times the upper limit of normal (ULN) in 1% or less. In trials of olaparib in patients with various advanced solid tumors there were no reports of hepatitis with jaundice or liver failure. Subsequent to its approval and more widescale use, there have been no published reports of clinically apparent liver injury attributed to olaparib.
Likelihood score: E (unlikely cause of clinically apparent liver injury).

---

Effects During Pregnancy and Lactation
◉ Summary of Use during Lactation
No information is available on the clinical use of olaparib during breastfeeding. Because olaparib is 82% bound to plasma proteins, the amount in milk is likely to be low. The manufacturer recommends that breastfeeding be discontinued during olaparib therapy and for one month after the last dose.
◉ Effects in Breastfed Infants
Relevant published information was not found as of the revision date.
◉ Effects on Lactation and Breastmilk
Relevant published information was not found as of the revision date.

---

Protein Binding
The protein binding of olaparib is approximately 82% _in vitro_. In solutions of purified proteins, the olaparib fraction bound to albumin was approximately 56% and the fraction bound to alpha-1 acid glycoprotein was 29%.

[1]. J Med Chem . 2008 Oct 23;51(20):6581-91.

[2]. Clin Cancer Res . 2008 Jun 15;14(12):3916-25.

[3]. Proc Natl Acad Sci U S A . 2008 Nov 4;105(44):17079-84.

[3]. Blood . 2010 Nov 25;116(22):4578-87.

Olaparib is a member of the class of N-acylpiperazines obtained by formal condensation of the carboxy group of 2-fluoro-5-[(4-oxo-3,4-dihydrophthalazin-1-yl)methyl]benzoic acid with the free amino group of N-(cyclpropylcarbonyl)piperazine; used to treat advanced ovarian cancer. It has a role as an antineoplastic agent, an EC 2.4.2.30 (NAD(+) ADP-ribosyltransferase) inhibitor and an apoptosis inducer. It is a N-acylpiperazine, a member of cyclopropanes, a member of monofluorobenzenes and a member of phthalazines.
Olaparib is a selective and potent inhibitor of poly (ADP-ribose) polymerase (PARP) enzymes, PARP1 and PARP2. PARP inhibitors represent a novel class of anti-cancer therapy and they work by taking advantage of a defect in DNA repair in cancer cells with BRCA mutations and inducing cell death. Olaparib is used to treat a number of BRCA-associated tumours, including ovarian cancer, breast cancer, pancreatic cancer, and prostate cancer. It was first approved by the FDA and EU in December 2014, and by Health Canada in April 2016.
Olaparib is a Poly(ADP-Ribose) Polymerase Inhibitor. The mechanism of action of olaparib is as a Poly(ADP-Ribose) Polymerase Inhibitor.
Olaparib is a small molecule inhibitor of poly ADP-ribose polymerase and is used as an antineoplastic agent in the therapy of refractory and advanced ovarian carcinoma. Olaparib therapy is associated with a low rate of transient elevations in serum aminotransferase during therapy and has not been linked to instances of clinically apparent liver injury.
Olaparib is a small molecule inhibitor of the nuclear enzyme poly(ADP-ribose) polymerase (PARP) with potential chemosensitizing, radiosensitizing, and antineoplastic activities. Olaparib selectively binds to and inhibits PARP, inhibiting PARP-mediated repair of single strand DNA breaks; PARP inhibition may enhance the cytotoxicity of DNA-damaging agents and may reverse tumor cell chemoresistance and radioresistance. PARP catalyzes post-translational ADP-ribosylation of nuclear proteins and can be activated by single-stranded DNA breaks.

---

Drug Indication
**Ovarian cancer** Olaparib is indicated for the maintenance treatment of adults with deleterious or suspected deleterious germline or somatic BRCA-mutated advanced epithelial ovarian, fallopian tube or primary peritoneal cancer who are in complete or partial response to first-line platinum-based chemotherapy. Olaparib is indicated in combination with [bevacizumab] for the maintenance treatment of adults with advanced epithelial ovarian, fallopian tube or primary peritoneal cancer who are in complete or partial response to first-line platinum-based chemotherapy and whose cancer is associated with homologous recombination deficiency (HRD)-positive status defined by either: a deleterious or suspected deleterious BRCA mutation, and/or genomic instability. Olaparib is indicated for the maintenance treatment of adult patients with recurrent epithelial ovarian, fallopian tube or primary peritoneal cancer, who are in complete or partial response to platinum-based chemotherapy. **Breast cancer** Olaparib is indicated for the adjuvant treatment of adult patients with deleterious or suspected deleterious g_BRCA_m human epidermal growth factor receptor 2 (HER2)-negative high risk early breast cancer who have been treated with neoadjuvant or adjuvant chemotherapy. Olaparib is indicated for the treatment of adult patients with deleterious or suspected deleterious g_BRCA_m, HER2-negative metastatic breast cancer, who have been treated with chemotherapy in the neoadjuvant, adjuvant, or metastatic setting. Patients with hormone receptor (HR) positive breast cancer should have been treated with a prior endocrine therapy or be considered inappropriate for endocrine therapy. **Pancreatic cancer** Olaparib is indicated for the maintenance treatment of adult patients with deleterious or suspected deleterious gBRCAm metastatic pancreatic adenocarcinoma whose disease has not progressed on at least 16 weeks of a first-line platinum-based chemotherapy regimen. **Prostate cancer** Olaparib is indicated for the treatment of adult patients with deleterious or suspected deleterious germline or somatic homologous recombination repair (HRR) gene-mutated metastatic castration-resistant prostate cancer (mCRPC) who have progressed following prior treatment with a hormone agent, such as [enzalutamide] or [abiraterone]. It is also indicated in combination with [abiraterone] and [prednisone] or [prednisolone] for the treatment of adult patients with deleterious or suspected deleterious BRCA-mutated (BRCAm) metastatic castration-resistant prostate cancer (mCRPC).
Ovarian cancer Lynparza is indicated as monotherapy for the: maintenance treatment of adult patients with advanced (FIGO stages III and IV) BRCA1/2-mutated (germline and/or somatic) high-grade epithelial ovarian, fallopian tube or primary peritoneal cancer who are in response (complete or partial) following completion of first-line platinum-based chemotherapy. maintenance treatment of adult patients with platinum sensitive relapsed high grade epithelial ovarian, fallopian tube, or primary peritoneal cancer who are in response (complete or partial) to platinum based chemotherapy. Lynparza in combination with bevacizumab is indicated for the: maintenance treatment of adult patients with advanced (FIGO stages III and IV) high-grade epithelial ovarian, fallopian tube or primary peritoneal cancer who are in response (complete or partial) following completion of first-line platinum-based chemotherapy in combination with bevacizumab and whose cancer is associated with homologous recombination deficiency (HRD) positive status defined by either a BRCA1/2 mutation and/or genomic instability (see section 5. 1). Breast cancer Lynparza is indicated as: monotherapy or in combination with endocrine therapy for the adjuvant treatment of adult patients with germline BRCA1/2-mutations who have HER2-negative, high risk early breast cancer previously treated with neoadjuvant or adjuvant chemotherapy (see sections 4. 2 and 5. 1). monotherapy for the treatment of adult patients with germline BRCA1/2-mutations, who have HER2 negative locally advanced or metastatic breast cancer . Patients should have previously been treated with an anthracycline and a taxane in the (neo)adjuvant or metastatic setting unless patients were not suitable for these treatments (see section 5. 1). Patients with hormone receptor (HR)-positive breast cancer should also have progressed on or after prior endocrine therapy, or be considered unsuitable for endocrine therapy. Adenocarcinoma of the pancreasLynparza is indicated as: monotherapy for the maintenance treatment of adult patients with germline BRCA1/2-mutations who have metastatic adenocarcinoma of the pancreas and have not progressed after a minimum of 16 weeks of platinum treatment within a first-line chemotherapy regimen. Prostate cancer Lynparza is indicated as: monotherapy for the treatment of adult patients with metastatic castration-resistant prostate cancer (mCRPC) and BRCA1/2-mutations (germline and/or somatic) who have progressed following prior therapy that included a new hormonal agent. in combination with abiraterone and prednisone or prednisolone for the treatment of adult patients with mCRPC in whom chemotherapy is not clinically indicated (see section 5. 1).
Treatment of all conditions included in the category of malignant neoplasms (except central nervous system tumours, haematopoietic and lymphoid tissue neoplasms)

---

Mechanism of Action
Poly(ADP-ribose) polymerases (PARPs) are multifunctional enzymes comprising 17 members. They are involved in essential cellular functions, such as DNA transcription and DNA repair. PARPs recognize and repair cellular DNA damage, such as single-strand breaks (SSBs) and double-strand breaks (DSBs). Different DNA repair pathways exist to repair these DNA damages, including the base excision repair (BER) pathway for SSBs and BRCA-dependent homologous recombination for DSBs. Olaparib is a PARP inhibitor: while it acts on PARP1, PARP2, and PARP3, olaparib is a more selective competitive inhibitor of NAD⁺ at the catalytic site of PARP1 and PARP2. Inhibition of the BER pathway by olaparib leads to the accumulation of unrepaired SSBs, which leads to the formation of DSBs, which is the most toxic form of DNA damage. While BRCA-dependent homologous recombination can repair DSBs in normal cells, this repair pathway is defective in cells with BRCA1/2 mutations, such as certain tumour cells. Inhibition of PARP in cancer cells with BRCA mutations leads to genomic instability and apoptotic cell death. This end result is also referred to as synthetic lethality, a phenomenon where the combination of two defects - inhibition of PARP activity and loss of DSB repair by HR - that are otherwise benign when alone, lead to detrimental results. _In vitro_ studies have shown that olaparib-induced cytotoxicity may involve inhibition of PARP enzymatic activity and increased formation of PARP-DNA complexes, resulting in DNA damage and cancer cell death.

C24H23FN4O3

434.46

434.175

C, 66.35; H, 5.34; F, 4.37; N, 12.90; O, 11.05

763113-22-0

23725625

White solid powder

1.4±0.1 g/cm3

1.702

1.9

86.37

1

5

4

32

790

0

FC1C([H])=C([H])C(C([H])([H])C2C3=C([H])C([H])=C([H])C([H])=C3C(N([H])N=2)=O)=C([H])C=1C(N1C([H])([H])C([H])([H])N(C([H])([H])C1([H])[H])C(C1([H])C([H])([H])C1([H])[H])=O)=O

FDLYAMZZIXQODN-UHFFFAOYSA-N

InChI=1S/C24H23FN4O3/c25-20-8-5-15(14-21-17-3-1-2-4-18(17)22(30)27-26-21)13-19(20)24(32)29-11-9-28(10-12-29)23(31)16-6-7-16/h1-5,8,13,16H,6-7,9-12,14H2,(H,27,30)

4-[[3-[4-(cyclopropanecarbonyl)piperazine-1-carbonyl]-4-fluorophenyl]methyl]-2H-phthalazin-1-one

AZD2281; Ku-0059436; AZD2281; AZD-2281; AZD 2281; KU59436; KU-59436; KU 59436; KU0059436; KU-0059436; KU 0059436; Olaparib; trade name Lynparza

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

配方 1 中的溶解度: 10 mg/mL (23.02 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (这些助溶剂从左到右依次添加，逐一添加), 悬浮液；超声助溶。
例如，若需制备1 mL的工作液，可将100 μL 100.0 mg/mL澄清DMSO储备液加入400 μL PEG300中，混匀；然后向上述溶液中加入50 μL Tween-80，混匀；加入450 μL生理盐水定容至1 mL。
*生理盐水的制备：将 0.9 g 氯化钠溶解在 100 mL ddH₂O中，得到澄清溶液。

---

配方 2 中的溶解度: ≥ 5 mg/mL (11.51 mM) (饱和度未知) in 5% DMSO + 40% PEG300 + 5% Tween80 + 50% Saline (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
*生理盐水的制备：将 0.9 g 氯化钠溶解在 100 mL ddH₂O中，得到澄清溶液。

---

View More

配方 3 中的溶解度: ≥ 5 mg/mL (11.51 mM) (饱和度未知) in 5% DMSO + 95% (20% SBE-β-CD in Saline) (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
*20% SBE-β-CD 生理盐水溶液的制备（4°C，1 周）：将 2 g SBE-β-CD 溶解于 10 mL 生理盐水中，得到澄清溶液。

---

配方 4 中的溶解度: ≥ 2.5 mg/mL (5.75 mM) (饱和度未知) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将100μL 25.0mg/mL澄清的DMSO储备液加入到900μL 20％SBE-β-CD生理盐水中，混匀。
*20% SBE-β-CD 生理盐水溶液的制备（4°C，1 周）：将 2 g SBE-β-CD 溶解于 10 mL 生理盐水中，得到澄清溶液。

---

配方 5 中的溶解度: ≥ 2.5 mg/mL (5.75 mM) (饱和度未知) in 10% DMF 40% PEG300 + 5% Tween80 + 45% Saline (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
*生理盐水的制备：将 0.9 g 氯化钠溶解在 100 mL ddH₂O中，得到澄清溶液。

---

配方 6 中的溶解度: ≥ 2.5 mg/mL (5.75 mM) (饱和度未知) in 10% DMF 90% (20% SBE-β-CD in Saline) (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
*20% SBE-β-CD 生理盐水溶液的制备（4°C，1 周）：将 2 g SBE-β-CD 溶解于 10 mL 生理盐水中，得到澄清溶液。

---

配方 7 中的溶解度: ≥ 2.5 mg/mL (5.75 mM) (饱和度未知) in 10% DMF 90% Corn Oil (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。

---

配方 8 中的溶解度: ≥ 2.08 mg/mL (4.79 mM) (饱和度未知) in 10% DMSO + 90% Corn Oil (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，要配制1 mL工作液，可将100 μL 20.8 mg/mL 澄清DMSO 储备液加入900 μL 玉米油中，混匀。

---

配方 9 中的溶解度: ≥ 0.5 mg/mL (1.15 mM) (饱和度未知) in 1% DMSO 99% Saline (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
*生理盐水的制备：将 0.9 g 氯化钠溶解在 100 mL ddH₂O中，得到澄清溶液。

---

配方 10 中的溶解度: 20 mg/mL (46.03 mM) in 0.5% CMC-Na/saline water (这些助溶剂从左到右依次添加，逐一添加), 悬浊液; 超声助溶。
*生理盐水的制备：将 0.9 g 氯化钠溶解在 100 mL ddH₂O中，得到澄清溶液。

---

请根据您的实验动物和给药方式选择适当的溶解配方/方案：
1、请先配制澄清的储备液(如：用DMSO配置50 或 100 mg/mL母液(储备液));
2、取适量母液，按从左到右的顺序依次添加助溶剂，澄清后再加入下一助溶剂。以 下列配方为例说明 (注意此配方只用于说明，并不一定代表此产品 的实际溶解配方):
10% DMSO → 40% PEG300 → 5% Tween-80 → 45% ddH2O (或 saline);
假设最终工作液的体积为 1 mL, 浓度为5 mg/mL: 取 100 μL 50 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀/澄清；向上述体系中加入50 μL Tween-80，混合均匀/澄清；然后继续加入450 μL ddH2O (或 saline)定容至 1 mL；
3、溶剂前显示的百分比是指该溶剂在最终溶液/工作液中的体积所占比例;
4、 如产品在配制过程中出现沉淀/析出，可通过加热(≤50℃)或超声的方式助溶;
5、为保证最佳实验结果，工作液请现配现用！
6、如不确定怎么将母液配置成体内动物实验的工作液，请查看说明书或联系我们;
7、 以上所有助溶剂都可在 Invivochem.cn网站购买。