Desidustat   中文名称: 德杜司他

HIF hydroxylase

提高成绩的物质和方法已成为竞技体育中的一个严重问题。缺氧诱导因子（HIF）稳定剂可以增强生物体的分子氧运输能力，并可能被滥用为运动中的性能增强剂。本文描述了使用马肝微粒体在QExactive高分辨率质谱仪上测定的流行缺氧诱导因子脯氨酰羟化酶（HIF-PH）抑制剂，即daprodustat、Desidustat和vadadustat的代谢转化。在这项研究中，共检测到10种达布达司他代谢物（均为I期）、10种Desidustat （I期和II期各5种）和15种vadadustat代谢物（I期6种，II期9种）。当前研究的重要发现如下：（1）所有三种HIF-PH抑制剂候选药物都容易氧化，从而产生相应的羟基化代谢物；（2） 在Desidustat 中，还观察到肟键的水解和解离；（3） 观察到母体药物的葡糖醛酸结合物（达普司他除外）以及单羟基类似物；（4） 仅观察到vadadustat的磺酸结合代谢物[1]。

在体内，desindusat（口服；10-100 mg/kg）表现出良好的有效性[1]。
慢性肾病（CKD）与激活的炎症反应有关。Desidustat是一种脯氨酰羟化酶（PHD）抑制剂，可用于治疗与CKD相关的贫血，但其对CKD炎症和纤维化变化的影响尚未得到评估。在这项研究中，我们研究了地塞米松对急性和慢性肾损伤临床前模型中炎症和纤维化变化的影响。通过缺血再灌注在雄性Sprague-Dawley大鼠中诱导急性肾损伤，其中评估了去沙尘暴（15mg/kg，PO）的影响。在另一项实验中，雄性C57小鼠用腺嘌呤处理14天以诱导CKD。这些小鼠接受去沙司他（15mg/kg，口服，隔日一次）治疗14天，并继续服用腺嘌呤。去沙司他可预防血清肌酐、尿素、IL-1β、IL-6和肾损伤分子-1（KIM-1）的升高，并提高急性肾损伤大鼠的促红细胞生成素水平。用腺嘌呤治疗的小鼠会出现CKD和贫血，去粉尘治疗可改善血清肌酐、尿素，还可改善血红蛋白，降低肝脏和血清hepcidin。脱尘处理可显著降低IL-1β、IL-6、髓过氧化物酶（MPO）和氧化应激。通过组织学分析和羟脯氨酸含量观察到，去沙司他治疗也减少了肾纤维化。脱尘治疗减少了肾纤维化和炎症，同时减少了肾损伤临床前模型中的贫血，这可能转化为对CKD患者的保护作用[1]

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自身免疫性溶血性贫血（AIHA）是一组由针对红细胞的自身抗体介导的异质性疾病，导致溶血和贫血。根据触发因素，AIHA会迅速或随着时间的推移而发展。Desidustat是一种脯氨酰羟化酶抑制剂，临床上用于治疗慢性肾病（CKD）引起的贫血。在这项研究中，我们研究了desidustat在AIHA临床前模型中的作用。我们使用大鼠红细胞诱导小鼠AIHA。然后用去沙司他（15mg/kg，口服，每天一次）治疗这些小鼠八周。去尘治疗增加了血红蛋白、红细胞和红细胞压积，降低了白细胞和淋巴细胞。这种治疗抑制了血清LDH、红细胞氧化应激、抗体滴度和红细胞表面抗体沉积，延长了红细胞寿命。去铁可降低血清和脾脏铁含量，同时减轻脾脏重量和氧化应激。通过治疗去铁，骨髓铁含量增加，骨髓中CD71（早期红系祖细胞的细胞表面标志物）和TER-119（晚期红系祖动物的细胞表面标记物）的表达升高。这种治疗还抑制了膜结合抗体在晚期红系细胞中的沉积。治疗显示脾脏总细胞、CD71和TER-119阳性细胞减少。因此，去粉尘治疗增加了红细胞生成，骨髓红系细胞的早期成熟具有更长的红细胞寿命，这是由于抗体介导的红细胞及其祖细胞的裂解减少，导致氧化应激减少。因此，desidustat可以成为治疗AIHA的良好治疗选择[2]。

在体外，检查了Caco2细胞通透性、血浆蛋白结合、代谢、细胞色素P450（CYP）抑制和CYP诱导[4]。

Animal/Disease Models: C57 mice[1]
Doses: 10, 30, 50, 100 mg/kg; 20 mg/kg
Route of Administration: po (oral gavage); oral administration, one time/day for 7 days.
Experimental Results: EPO and Hb levels demonstrated significant rising.

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Acute kidney injury in SD rats [1]
Male SD rats (age 7–8 weeks, and body weight 210–240 g) were anesthetized with ketamine (50 mg/kg, IP) and xylazine (10 mg/kg, IP) and both the renal pedicles were exposed after laparotomy. Bilateral renal pedicles were occluded with hemostasis clamp for 25 min. At the end of bilateral ischemia, the clips were removed to allow reperfusion of blood in kidney. The skin and muscle were closed by surgical suture 3.0. Vehicle in ischemia reperfusion injury (I/R) group (n = 9) or Desidustat (15 mg/kg, PO, n = 9) were administered 30 min prior to and 2 h after to initiation of renal ischemia. In sham (normal) control animals only renal pedicles were exposed after laparotomy. After 24 h of reperfusion, all the animals were bled retro-orbitally under isoflurane anesthesia for collection of blood. Serum samples were analyzed for creatinine, urea, IL-1β, IL-6, erythropoietin, and kidney injury molecule, KIM-1 levels using assay kits mentioned in parentheses.

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Chronic kidney disease in C57 mice [1]
Chronic kidney disease was induced in C57 mice (age 7–8 weeks, and body weight 25–30 g) by adenine supplementation. Animals were randomized based on their body weight into two groups; Adenine group (n = 8), and Desidustat (15 mg/kg, n = 8). All these groups were fed with adenine at 50 mg/kg, by oral route (PO) for 14 days, and for next 14 days they were given Desidustat treatment on alternate day for 14 days, with adenine supplementation continued. A chow-fed control group was separately maintained. On 15th day, the animals were kept in metabolic cages for collection of urine. Next day, animals were bled retro-orbitally under isoflurane anesthesia and serum was separated. Albumin, creatinine, and urea were also measured using Labcare Diagnostics kits. Estimated glomerular filtration rate was calculated as described in literature (Pestel, Krzykalla, & Weckesser, 2007). IL-1β, IL-6 levels using assay kits mentioned in parentheses. Hepcidin in serum and liver were measured using ELISA kit. Lipid peroxidation products, in terms of thiobarbituric reacting substances (TBARS), were measured in kidneys collected immediately after sacrifice, and stored at −70°C until the assay. TBARS was assessed by a spectrophotometric method (Buege, & Aust, 1978). Lipid peroxidation was expressed in terms of malondialdehyde (MDA) equivalents using an extinction coefficient of 1.56 X 105 L / mol per cm and results are expressed as nmol MDA / g tissue. Superoxide dismutase (SOD) activity in kidney was measured spectrophotometrically by the inhibition of pyrogallol auto-oxidation at 420 nm for 10 min (Marklund, & Marklund, 1974). One unit activity was determined as the amount of enzyme that inhibited the oxidation of pyrogallol by 50%. Myeloperoxidase (MPO) was measured in the serum and kidneys (Kim et al., 2012). In addition, kidneys were also processed for the determination of hydroxyproline using Quickzyme hydroxyproline assay kit.

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Induction of autoimmune hemolytic anemia [2]
Autoimmune hemolytic anemia (AIHA) was induced in mice by repeated injections of rat erythrocytes (RBCs). SD rats were bled by retroorbital puncture under isoflurane anesthesia in a tube containing disodium EDTA as an anticoagulant. Plasma was removed and cells were washed three times with phosphate-buffered saline (PBS, pH 7.4) and cells adjusted to a concentration of 1 × 109 cells/ml in PBS. Mice were given injections of approximately 2 × 108 rat RBCs by intraperitoneal route once in a week. After six weeks of initiation of weekly rat RBC injections, mice blood was collected from the tail vein for estimation of complete blood count and were randomized based on RBC, and hemoglobin content. The groups were vehicle control (10 mL/kg), or Desidustat (15 mg/10 mL/kg, once a day, PO) treatment for next eight weeks. Injection of rat RBC was also continued with this treatment for next eight weeks. At the end of treatment, whole blood (for serum and complete blood count), bone marrow, liver and spleen were collected. Lactate dehydrogenase activity was measured in serum samples using a kit in Cobas 6000 instrument.

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In vivo, pharmacokinetic studies of oral bioavailability in mice, rats, dogs and monkeys, dose linearity, tissue distribution, and excretion in rats were conducted.[4]

In Caco-2 cells, the apparent permeability of desidustat was high at low pH and low at neutral pH. The oral bioavailability (%F) of desidustat was 43–100% with a median time to reach peak concentration (Tmax) of about 0.25–1.3 h across species. Desidustat displayed a low mean plasma clearance (CL) of 1.3–4.1 mL/min/kg (approximately 1.8–7.4% of hepatic blood flow), and the mean steady-state volume of distribution (Vss) was 0.2–0.4 L/kg (approximately 30–61% of the total body water). Desidustat showed a dose-dependent increase in exposures over the 15–100 mg/kg dose range. It was rapidly distributed in various tissues, with the highest tissue-to-blood ratio in the liver (1.8) and kidney (1.7). Desidustat showed high plasma protein binding and was metabolically stable in human liver microsomes, hepatocytes, and recombinant CYPs. It did not show significant inhibition of major drug-metabolizing CYP enzymes (IC50 > 300 µM) or the potential to induce CYP1A2 and CYP3A4/5 (up to 100 µM) in HepG2 cells. It may have minimal potential of clinical drug–drug interaction when used in combination with iron supplements or phosphate binders. Desidustat was primarily excreted unchanged in urine (25% of the oral dose) and bile (25% of the oral dose) in rats. The mean elimination half-life of desidustat ranged from 1.0 to 5.3 h and 1.3 to 5.7 h across species after intravenous and oral administration, respectively. [4]
Taken together, desidustat is well absorbed orally. It showed a dose-dependent increase in exposure, did not accumulate in tissue, and was eliminated via dual routes. It is metabolically stable, has minimal potential to cause clinical drug–drug interactions (DDIs), and demonstrates discriminable pharmacokinetic properties for the treatment of anemia.[4]

[1]. Novel quinolone derivatives. Patent. WO2014102818A1.

[2]. Prolyl hydroxylase inhibitor desidustat attenuates autoimmune hemolytic anemia in mice. Int Immunopharmacol. 2024 Dec 5;142(Pt A):113029.

[3]. In vitro studies of hypoxia inducible factor-prolyl hydroxylase inhibitors daprodustat, desidustat, and vadadustat for equine doping control. Drug Test Anal. 2022 Feb;14(2):317-348.

[4]. Nonclinical Pharmacokinetic Evaluation of Desidustat: a Novel Prolyl Hydroxylase Inhibitor for the Treatment of Anemia. Eur J Drug Metab Pharmacokinet. 2022 Sep;47(5):725-740.

Desidustat is a N-acyl-amino acid.
Desidustat is under investigation in clinical trial NCT04012957 (Desidustat in the Treatment of Anemia in CKD).
Desidustat is an orally bioavailable, hypoxia-inducible factor prolyl hydroxylase (HIF-PH) inhibitor (HIF-PHI), with potential anti-anemic and anti-inflammatory activities. Upon administration, desidustat binds to and inhibits HIF-PH, an enzyme responsible for the degradation of transcription factors in the HIF family under normal oxygen conditions. This prevents HIF breakdown and promotes HIF activity. Increased HIF activity leads to an increase in endogenous erythropoietin production, thereby enhancing erythropoiesis. It also reduces the expression of the peptide hormone hepcidin, improves iron availability, and boosts hemoglobin (Hb) levels. HIF regulates the expression of genes in response to reduced oxygen levels, including genes required for erythropoiesis and iron metabolism. In addition, HIF 1-alpha (HIF1A) may play a role in reducing inflammation during acute lung injury (ALI) through HIF-dependent control of glucose metabolism in the alveolar epithelium.

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Mechanism of Action
The small molecule hypoxia-inducible factor, desidustat, inhibits the prolyl hydrozylase and stimulates erythropoiesis. It is currently being investigated against anemia of inflammation and COVID-19.

C16H16N2O6

332.308044433594

332.1

C, 57.83; H, 4.85; N, 8.43; O, 28.89

1616690-16-4

1616690-16-4;

75593290

White to off-white solid powder

1.5±0.1 g/cm3

1.676

0.54

116

3

6

6

24

583

0

O=C(O)CNC(C1=C(O)C2=C(N(OCC3CC3)C1=O)C=CC=C2)=O

IKRKQQLJYBAPQT-UHFFFAOYSA-N

InChI=1S/C16H16N2O6/c19-12(20)7-17-15(22)13-14(21)10-3-1-2-4-11(10)18(16(13)23)24-8-9-5-6-9/h1-4,9,21H,5-8H2,(H,17,22)(H,19,20)

2-[[1-(cyclopropylmethoxy)-4-hydroxy-2-oxoquinoline-3-carbonyl]amino]acetic acid

Desidustat; 1616690-16-4; ZYAN1; Zyan-1; Desidustat [INN]; Oxemia; ZYAN1 compound; Y962PQA4KS;

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)

DMSO : ~10 mg/mL (~30.09 mM)

配方 1 中的溶解度: ≥ 1 mg/mL (3.01 mM) (饱和度未知) 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), 澄清溶液。
例如，若需制备1 mL的工作液，可将100 μL 10.0 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。

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配方 2 中的溶解度: ≥ 1 mg/mL (3.01 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，若需制备1 mL的工作液，可将 100 μL 10.0 mg/mL 澄清 DMSO 储备液加入到 900 μL 玉米油中并混合均匀。

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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。