曲美曲沙（0.1 μM，18 小时）完全抑制由弓形虫引起的小鼠巨噬细胞肿胀[3]。弓形虫细胞膜可渗透三甲曲酯 (1 μM)，可快速达到细胞内高浓度（108 pmol/107 个细胞）。在 SNU-C4 和 NCI-H630 细胞系中，曲美曲沙（0.1 mM；24 小时）可降低细胞生长 50-60% [5]。十分钟内，添加 C4 中的曲美曲沙（1 和 10 mM；24 小时）[3]。

Trimetrexate（180 mg/kg 或 30 mg/kg；口服或腹腔注射；每日）可延长弓形虫感染的中位生存期，并显示出抗弓形虫活性 [3]。曲美曲沙（0-30 mg/kg；静脉注射；每天一次，持续 5 天）表现出慢性毒性[4]。

细胞增殖测定[5]
细胞类型： SNU-C4 和 NCI-H630
测试浓度： 0.1 mM
孵育时间： 24小时
实验结果：两种细胞系均抑制细胞生长50-60%。

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细胞增殖测定[5]
细胞类型： C4 细胞
测试浓度： 1 和 10 mM
孵育持续时间：24 小时
实验结果：在 1 和 10 mM 浓度下分别产生 42% 和 50% 的致死率。

Animal/Disease Models: Female balb/c (Bagg ALBino) mouse infected with Toxoplasma gondii, weighing about 20 g [3]
Doses: 180 mg/kg or 30 mg/kg Dosing: 180 mg/kg orally daily in drinking water or intraperitoneal (ip) injection 30 times daily mg/kg
Experimental Results: Extended median survival of infected mice to 10 days (oral) or 19 days (ip).

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Animal/Disease Models: Charles River Wistar Crl(WI)BR rats, weighing approximately 150 to 200 g[4]
Doses: 0, 1, 10 or 30 mg/kg
Route of Administration: intravenous (iv) (iv)injection, one time/day for 5 days, then 23-
Experimental Results: Showing chronic toxicity, testicular changes that persisted over the course of multiple dosing cycles were irreversible within 21 days but required an additional 56 days for essentially complete recovery.

Absorption, Distribution and Excretion
Ten to 30% of the administered dose is excreted unchanged in the urine.
20 ± 8 L/m2
36.9 ± 6 L/m2 [cancer patients]
38 +/- 15 mL/min/m2 [patients with acquired immunodeficiency syndrome (AIDS) who had Pneumocystis carinii pneumonia (4 patients) or toxoplasmosis (2 patients). Trimetrexate was administered intravenously as a bolus injection at a dose of 30 mg/m2/day along with leucovorin 20 mg/m2 every 6 hours for 21 days]
53 +/- 41 mL/min/m2 [Cancer patients with advanced solid tumors using various dosage regimensreceiving a single-dose administration of 10 to 130 mg/m2]
30 +/- 8 mL/min/m2 [Cancer patients with advanced solid tumors using various dosage regimensafter a five-day infusion]
Clinical pharmacokinetic studies in cancer patients show that trimetrexate plasma concentration-time curves are biphasic or triphasic in form. The terminal elimination half-life averages 13.6 hr. Mean total plasma clearance and volume of distribution as steady-state values were 27.9 ml/min/sq m and 21.1 l/sq m, respectively. Cerebrospinal fluid concentration was 3.4% of that in plasma, which shows that trimetrexate does not cross the blood-brain barrier well. Trimetrexate is 86 to 94% bound to plasma proteins.
Mean oral bioavailability of the parenteral solution (glucuronate) in AIDS patients was 42%, with a mean maximum plasma concentration of 1182 ng/ml (3.2 umole/l) achieved 1.8 hr postdose. /Trimetrexate glucuromate/

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Metabolism / Metabolites
Hepatic. Preclinical data strongly suggest that the major metabolic pathway is oxidative O-demethylation, followed by conjugation to either glucuronide or the sulfate.
Trimetrexate is highly metabolized by the liver. At least 2 metabolites are excreted in urine. One metabolite has been identified a 4'-O-glucuronide conjugate of trimetrexate, which is formed as a result of oxidative O-dimethylation at the 4'-position and subsequent conjugation with glucuronic acid. About 15% of a dose is excreted in urine as unchanged trimetrexate, while another 20% apparently excreted as metabolites.
Hepatic. Preclinical data strongly suggest that the major metabolic pathway is oxidative O-demethylation, followed by conjugation to either glucuronide or the sulfate.
Route of Elimination: Ten to 30% of the administered dose is excreted unchanged in the urine.
Half Life: 11 to 20 hours

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Biological Half-Life
11 to 20 hours
The terminal elimination half-life averages 13.6 hours.

Toxicity Summary
In vitro studies have shown that trimetrexate is a competitive inhibitor of dihydrofolate reductase (DHFR) from bacterial, protozoan, and mammalian sources. DHFR catalyzes the reduction of intracellular dihydrofolate to the active coenzyme tetrahydrofolate. Inhibition of DHFR results in the depletion of this coenzyme, leading directly to interference with thymidylate biosynthesis, as well as inhibition of folate-dependent formyltransferases, and indirectly to inhibition of p.r.n. biosynthesis. The end result is disruption of DNA, RNA, and protein synthesis, with consequent cell death.

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Hepatotoxicity
When given without leucovorin protection, trimetrexate therapy is associated with a moderate rate of serum enzyme elevations, serum ALT or AST elevations above 5 times ULN in up to 20% of patients. When given with leucovorin, however, trimetrexate has fewer side effects although serum enzyme elevations can still occur. In clinical trials in patients with HIV infection and Pneumocystis jirovecii pneumonia, ALT elevations above 5 times ULN occurred in 1% to 8% of patients, but were usually no more frequent than with standard therapy using trimethoprim with sulfamethoxazole. The elevations were typically transient, without accompanying symptoms or jaundice and resolved or improved despite continuation of therapy. No instances of clinically apparent acute liver injury attributed to trimetrexate have been reported in the literature. In addition, trimetrexate has not been linked to sinusoidal obstruction syndrome or to reactivation of hepatitis B. Nevertheless, trimetrexate probably has hepatotoxic potential, but because it has limited use, is given for short periods of time and is administered with leucovorin, it has not been convincingly linked to cases of clinically apparent liver injury with jaundice.
Likelihood score: E* (unproven but suspected cause of liver injury).

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Protein Binding
95% (over the concentration range of 18.75 to 1000 ng/mL)

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Toxicity Data
LD50: 62 mg/kg (Intravenous, Mouse) (A308)

[1]. Hopper AT, et al. Discovery of Selective Toxoplasma gondii Dihydrofolate Reductase Inhibitors for the Treatment of Toxoplasmosis. J Med Chem. 2019 Feb 14;62(3):1562-1576.
[2]. Fulton, B., et al. Trimetrexate. Drugs 49, 563–576 (1995).
[3]. Allegra CJ, et al. Potent in vitro and in vivo antitoxoplasma activity of the lipid-soluble antifolate trimetrexate. J Clin Invest. 1987 Feb;79(2):478-82.
[4]. Dethloff LA, et al. Chronic toxicity of the anticancer agent trimetrexate in rats. Fundam Appl Toxicol. 1992 Jul;19(1):6-14.
[5]. Grem JL, Voeller DM, Geoffroy F, Horak E, Johnston PG, Allegra CJ. Determinants of trimetrexate lethality in human colon cancer cells. Br J Cancer. 1994 Dec;70(6):1075-84.

Trimetrexate is a member of quinazolines. It has a role as an antifungal drug.
A nonclassical folic acid inhibitor through its inhibition of the enzyme dihydrofolate reductase. It is being tested for efficacy as an antineoplastic agent and as an antiparasitic agent against pneumocystis pneumonia in AIDS patients. Myelosuppression is its dose-limiting toxic effect.
Trimetrexate is a parenterally administered folate antagonist that is used as a second line therapy for severe Pneumocystis jirovecii (previously carinii) pneumonia. Trimetrexate therapy has been associated with transient, mild serum enzyme elevations during therapy, but has not been convincingly linked to instances of acute, clinically apparent liver injury.
Trimetrexate is a methotrexate derivative with potential antineoplastic activity. Trimetrexate inhibits the enzyme dihydrofolate reductase, thereby preventing the synthesis of purine nucleotides and thymidylate, with subsequent inhibition of DNA and RNA synthesis. Trimetrexate also exhibits antiviral activity. (NCI04)
A nonclassical folic acid inhibitor through its inhibition of the enzyme dihydrofolate reductase. It is being tested for efficacy as an antineoplastic agent and as an antiparasitic agent against pneumocystis pneumonia in AIDS patients. Myelosuppression is its dose-limiting toxic effect. [PubChem]
A nonclassical folic acid inhibitor through its inhibition of the enzyme dihydrofolate reductase. It is being tested for efficacy as an antineoplastic agent and as an antiparasitic agent against PNEUMOCYSTIS PNEUMONIA in AIDS patients. Myelosuppression is its dose-limiting toxic effect.

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Drug Indication
For use, with concurrent leucovorin administration (leucovorin protection), as an alternative therapy for the treatment of moderate-to-severe Pneumocystis carinii pneumonia (PCP) in immunocompromised patients, including patients with the acquired immunodeficiency syndrome (AIDS). Also used to treat several types of cancer including colon cancer.
FDA Label

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Mechanism of Action
In vitro studies have shown that trimetrexate is a competitive inhibitor of dihydrofolate reductase (DHFR) from bacterial, protozoan, and mammalian sources. DHFR catalyzes the reduction of intracellular dihydrofolate to the active coenzyme tetrahydrofolate. Inhibition of DHFR results in the depletion of this coenzyme, leading directly to interference with thymidylate biosynthesis, as well as inhibition of folate-dependent formyltransferases, and indirectly to inhibition of p.r.n. biosynthesis. The end result is disruption of DNA, RNA, and protein synthesis, with consequent cell death.
Trimetrexate binds to dihydrofolate reductase and prevents the conversion of dihydrofolate to biologically active tetrahydrofolate. It inhibits nucleic acid synthesis as a result of antithymidylate and antipurine effects.

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Therapeutic Uses
Antifungal Agents; Antimetabolites; Antimetabolites, Antineoplastic; Folic Acid Antagonists
Antineoplastic
Trimetrexate glucuronate is an investigational drug that is available for treatment use ... in a hospital setting in qualifying patients with Pneumocystis carinii pneumonia and who have exhibited serious ... intolerance to both co-trimoxazole and pentamidine. /Trimetrexate glucuronate/

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Drug Warnings
Even though trimetrexate does not compete with the folate transport system for entry into cells, utilization of folates is reduced due to inhibition of dihydrofolate reductase. When trimetrexate is administered in high doses for the treatment of pneumocystis carinii pneumonia, leucovorin must be given concurrently with trimetrexate to reduce toxic effects on human tissues. It has been postulated that the absence of a folate transport system in pneumocystis carinii provides the opportunity for differential rescue of host tissue affecting antiprotozoal action.
Patients receiving trimetrexate should have frequent laboratory monitoring of hepatic and hematologic prameters, including serum alanine aminotransferase, serum aspartate aminotransferase, bilirubin, alkaline phosphatase, platelet count, and total and differential leukocyte counts. Although uncommon in AIDS patients receiving trimetrexate for pneumocystis carinii pneumonia, slight elevations of blood urea nitrogen and/or serum creatinine have been reported in patients receiving the drug for various cancers.
Isolated perfused rat liver has been used to study potential drug interactions with trimetrexate metabolism in vitro. Cimetidine, which inhibits oxidative drug metabolizing enzymes, decreased the clearance of trimetrexate to approximately one half of control values. Whether this occurs to a significant extent in vivo is unknown.

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Pharmacodynamics
Trimetrexate, a non-classical folate antagonist, is a synthetic inhibitor of the enzyme dihydrofolate reductase (DHFR). During DNA synthesis and cellular reproduction, folic acid is reduced to tetrahydrofolic acid by the enzyme folic acid reductase. By interfering with the reduction of folic acid, trimetrexate interferes with tissue cell reproduction. Generally, the most sensitive cells to the antimetabolite effect of trimetrexate are those cells which are most actively proliferating such as malignant cells, dermal epithelium, buccal and intestinal mucosa, bone marrow, fetal cells, and cells of the urinary bladder. Because the proliferation of cells in malignant tissues is greater than in most normal tissues, trimetrexate may impair the growth of the malignant tissues without causing irreversible damage to normal tissues. Due to very serious and potentially life-threatening side-effects of this drug, leucovorin must be co-administered for at least 72 hours after the last dose.

C19H23N5O3

369.41762

369.18

52128-35-5

Trimetrexate glucuronate;82952-64-5;Trimetrexate trihydrochloride;1658520-97-8;Trimetrexate isethionate;82935-04-4

5583

Typically exists as solid at room temperature

1.305g/cm3

647ºC at 760mmHg

215-217 °C
215 - 217 °C

345.1ºC

3.975

117.54

3

8

6

27

457

0

NC1=C(C(C)=C2CNC3=CC(OC)=C(OC)C(OC)=C3)C(C=C2)=NC(N)=N1

NOYPYLRCIDNJJB-UHFFFAOYSA-N

InChI=1S/C19H23N5O3/c1-10-11(5-6-13-16(10)18(20)24-19(21)23-13)9-22-12-7-14(25-2)17(27-4)15(8-12)26-3/h5-8,22H,9H2,1-4H3,(H4,20,21,23,24)

5-methyl-6-[(3,4,5-trimethoxyanilino)methyl]quinazoline-2,4-diamine

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 : ≥ 61.5 mg/mL (~166.48 mM)

配方 1 中的溶解度: ≥ 2.5 mg/mL (6.77 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 25.0 mg/mL澄清DMSO储备液加入到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: ≥ 2.5 mg/mL (6.77 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 25.0 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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配方 3 中的溶解度: 40 mg/mL (108.28 mM) in 50% PEG300 50% Saline (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。

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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。