CN1314456C - Medicament box of 2beta-[N,N'-di(2-mercaptoethyl) ethylene diamine] methyl-3 beta-(4- chlorphenyl) tropane - Google Patents

Abstract

The present invention relates to a technetium [[99m]<Tc>] labeling kit of 2 beta-[N, N'-double (2-mercaptoethyl) ethylene diamine] methyl-3 beta-(4-chlorophenyl)tropane (TRODAT-1 for short), which belongs to the technical field of a single photon emission computed tomography (SPECT) imaging agent. The present invention provides a kit prescription of TRODAT-1. When used for labeling [[99m]<Tc>]], the kit prepared by the prescription has the advantages of simple operation and stable labeling rate (larger than 90%). The kit is prepared from TRODAT-1, stannous salt, glucoheptonate, edetate, a pH buffering agent, etc. according to a certain composition. No identical international report for the prescription of the kit provided by the present invention is reported.

Description

Translated fromChinese

2β-[N，N’-双(2-巯乙基)乙撑二胺]甲基-3β-(4-氯苯基)托烷的药盒Kit of 2β-[N,N'-bis(2-mercaptoethyl)ethylenediamine]methyl-3β-(4-chlorophenyl)tropane

技术领域technical field

2β-[N，N’-双(2-巯乙基)乙撑二胺]甲基-3β-(4-氯苯基)托烷(以下简称：TRODAT-1)的锝[^99mTc]标记药盒，涉及TRODAT-1的锝[^99mTc]标记药盒配方和制备技术，属于单光子发射计算机断层(SPECT)显像剂技术领域。Technetium [^99m Tc ] labeling of 2β-[N,N'-bis(2-mercaptoethyl)ethylenediamine]methyl-3β-(4-chlorophenyl)tropane (hereinafter referred to as: TRODAT-1) The medicine box relates to the technetium [^99m Tc ] labeled medicine box formula and preparation technology of TRODAT-1, and belongs to the technical field of single photon emission computed tomography (SPECT) imaging agent.

背景技术Background technique

锝[^99mTc]-2β-[N，N’-双(2-巯乙基)乙撑二胺]甲基-3β-(4-氯苯基)托烷(^99mTc-TRODAT-1)是第一个锝[^99mTc]标记的成功用于人体的多巴胺转运蛋白显像剂，它能与脑内的多巴胺转运蛋白特异结合而浓聚于纹状体，提供清晰的图像，提示多巴胺能系统的变化信息，对有关的疾病特别是对帕金森病的早期诊断具有极佳的临床应用潜力。自1996年Kung H.F.首次报道以来，国内外已开展了大量药理与临床研究，显示了优良的临床应用前景。目前^99mTc-TRODAT-1已成功用于对帕金森病的早期诊断和病情评价，并在注意力缺陷多动综合症方面也取能了成功的应用。Technetium [^99m Tc ]-2β-[N,N'-bis(2-mercaptoethyl)ethylenediamine]methyl-3β-(4-chlorophenyl)tropane (^99m Tc-TRODAT-1) is The first technetium [^99m Tc ]-labeled dopamine transporter imaging agent successfully used in human body, it can specifically combine with the dopamine transporter in the brain and concentrate in the striatum, providing clear images, suggesting the dopaminergic system The change information has excellent clinical application potential for related diseases, especially for the early diagnosis of Parkinson's disease. Since Kung HF was first reported in 1996, a large number of pharmacological and clinical studies have been carried out at home and abroad, showing excellent clinical application prospects. At present,^99m Tc-TRODAT-1 has been successfully used in the early diagnosis and condition evaluation of Parkinson's disease, and has also been successfully applied in attention deficit hyperactivity syndrome.

临床应用的^99mTc-TRODAT-1要求标记过程简单、标记率最好大于90％。1997年Kung M.P等应用多步法制备^99mTc-TRODAT-1，虽然标记率大于90％，但操作繁琐，不能推广就用。为了便于临床应用，最好将制备^99mTc-TRODAT-1所需的TRODAT-1、还原剂、赋形剂等制成药盒，简化标记过程。为此，Choi S.R.等按10μg的TRODAT-1、32μg的氯化亚锡、10mg葡庚糖酸钠和840μg的乙二胺四乙酸二钠的配方制备了冻干药盒，建立了一步法标记方案，平均标记率90％。我们和其它研究人员都发现该配方的药盒缺点在于标记率不稳定，经常低于90％。为了解决这一缺点，Gyula Toth等将自制的药盒标记后，将标记溶液通过一个C₁₈小柱，再用洗脱剂按一定程序淋洗，除去大部份杂质，使放化纯大于95％，提供了一个制备高放化纯^99mTc-TRODAT-1的方法。但此方法增多了操作步骤，操作过程复杂，不利于推广，同时还增加了成本。The clinical application of^99m Tc-TRODAT-1 requires that the labeling process is simple and the labeling rate is preferably greater than 90%. In 1997, Kung MP et al. used a multi-step method to prepare^99m Tc-TRODAT-1. Although the labeling rate was greater than 90%, the operation was cumbersome and could not be popularized. For the convenience of clinical application, it is better to prepare TRODAT-1, reducing agent, excipients, etc. required for the preparation of^99m Tc-TRODAT-1 into a kit to simplify the labeling process. To this end, Choi SR et al. prepared a lyophilized kit according to the formula of 10 μg of TRODAT-1, 32 μg of stannous chloride, 10 mg of sodium glucoheptonate and 840 μg of disodium edetate, and established a one-step labeling method. program, with an average labeling rate of 90%. We and other researchers have found that the disadvantage of the formulation kit is that the labeling rate is not stable, often below 90%. In order to solve this shortcoming, after Gyula Toth et al. mark the self-made medicine box, pass the labeling solution through a C₁₈ small column, then rinse with an eluent according to a certain procedure, remove most of the impurities, and make the radiochemical purity greater than 95 %, a method for preparing highly radiochemically pure^99m Tc-TRODAT-1 is provided. However, this method increases the operation steps, and the operation process is complicated, which is not conducive to popularization, and also increases the cost.

发明内容Contents of the invention

本发明的目的是提供一种TRODAT-1标记药盒配方，使得标记率稳定并大于90％。为了克服国外TRODAT-1的标记药盒的缺点，我们设计了一种新的药盒配方，在药盒中添加了pH缓冲剂，并提高了TRODAT-1的用量，使得标记率稳定并大于90％，以利于临床应用，并且操作过程简单，便于推广。The purpose of the present invention is to provide a TRODAT-1 labeling kit formula, so that the labeling rate is stable and greater than 90%. In order to overcome the shortcomings of foreign TRODAT-1 labeling kits, we designed a new kit formula, added a pH buffer to the kit, and increased the amount of TRODAT-1, so that the labeling rate was stable and greater than 90 %, in order to facilitate clinical application, and the operation process is simple and easy to popularize.

本发明的技术方案：100瓶TRODAT-1药盒的制备方法如下，称取pH缓冲剂7mmol，溶于70mL蒸馏水中，再加入0.1-2.0g的葡庚糖酸盐和0.05-1.00g的乙二胺四乙酸盐，加入1mg/mL的亚锡盐稀盐酸(0.05mol/L)溶液0.4-10mL，最后加入1mg/mL的TRODAT-1稀盐酸(0.05mol/L)溶液2-20mL，再补加蒸馏水至药液总体积为100ml，再按1.0mL/瓶分装于10ml规格的西林瓶中，冷冻干燥后即得TRODAT-1药盒。每支药盒中含20-200μg的TRODAT-1、4-100μg的亚锡盐、0.5-10mg的乙二胺四乙酸盐和1-20mg的葡庚糖酸盐。药盒加1ml蒸馏水后pH值范围为4.0-8.5。Technical scheme of the present invention: the preparation method of 100 bottles of TRODAT-1 medicine box is as follows, take pH buffering agent 7mmol, be dissolved in 70mL distilled water, add the glucoheptonate of 0.1-2.0g and the ethyl alcohol of 0.05-1.00g Diamine tetraacetate, add 0.4-10mL of 1mg/mL stannous salt dilute hydrochloric acid (0.05mol/L) solution, and finally add 2-20mL of 1mg/mL TRODAT-1 dilute hydrochloric acid (0.05mol/L) solution, Then distilled water was added until the total volume of the medicinal solution was 100ml, and then 1.0mL/bottle was divided into 10ml vials, and the TRODAT-1 kit was obtained after freeze-drying. Each kit contains 20-200 μg of TRODAT-1, 4-100 μg of stannous salt, 0.5-10 mg of edetate and 1-20 mg of glucoheptonate. After adding 1ml of distilled water to the kit, the pH range is 4.0-8.5.

标记过程：取TRODAT-1药盒1支，加入Na^99mTcO₄溶液1mL，置于沸水浴上加热30min，冷却后即可。Labeling process: Take one TRODAT-1 kit, add 1mL of Na^99m TcO₄ solution, place it on a boiling water bath and heat for 30min, then cool it down.

标记率的测定方法为上行薄层色谱(TLC)法。The detection method of the labeling rate was the ascending thin-layer chromatography (TLC) method.

我们还考察了药盒中各组份含量在一定范围内变化时对药盒质量的影响，考察方法为保持其它组份的含量不变，在一定范围内改变一种成份的含量，进行锝[^99mTc]标记后测定标记率。结果表明当药盒中的组份含量在下列范围内，均可保证标记率大于90％：We also investigated the impact on the quality of the medicine box when the content of each component in the medicine box changes within a certain range. The investigation method is to keep the content of other components unchanged, change the content of one component within a certain range, and conduct technetium[^99m Tc] labeling to measure the labeling rate. The results show that when the content of the components in the kit is within the following ranges, the labeling rate can be guaranteed to be greater than 90%:

TRODAT-1          20-200μgTRODAT-1 20-200μg

pH值              4.0-8.5pH value 4.0-8.5

亚锡盐            4-100μgStannous salt 4-100μg

乙二胺四乙酸盐    0.5-10mgEDTA 0.5-10mg

葡庚糖酸盐        1-20mgGlucoheptonate 1-20mg

所用亚锡盐为氯化亚锡、氟化亚锡、酒石酸亚锡、溴化亚锡或柠檬酸亚锡。The stannous salt used is stannous chloride, stannous fluoride, stannous tartrate, stannous bromide or stannous citrate.

所用乙二胺四乙酸盐为乙二胺四乙酸钠盐或乙二胺四乙酸钾盐。The EDTA salt used is EDTA sodium salt or EDTA potassium salt.

所用葡庚糖酸盐为葡庚糖酸钠或葡庚糖酸钾。The glucoheptonate used was sodium glucoheptonate or potassium glucoheptonate.

药盒中pH缓冲剂是磷酸盐、柠檬酸盐、铵盐、醋酸盐、酒石酸盐、碳酸盐、硼酸盐、巴比妥盐、邻苯二钾酸盐中的一种或一种以上的混合物。The pH buffer in the kit is one or one of phosphate, citrate, ammonium salt, acetate, tartrate, carbonate, borate, barbiturate, and phthalate a mixture of the above.

本发明的有益效果：我们认为，造成Choi S设计的药盒标记率不稳定的原因是：(1)体系中不含pH缓冲剂，使得标记过程中pH值不能维持在固定值；(2)TRODAT-1用量过少。由于TRODAT-1化学性质不稳定，在制备、运输、贮藏过程中可能有部份甚至全部TRODAT-1发生分解变质。本发明针对性地进行了改进：(1)配方中添加了pH缓冲剂，使标记过程中pH值能维持在稳定值，标记率稳定并大于90％，十分有利于临床用药；(2)提高了药盒中的TRODAT-1用量，当有少量的TRODAT-1发生变质时，药盒中仍有足够量的TRODAT-1，保证标记率大于90％。本药盒还具有标记过程简单，可以方便地进行推广应用的特点。本发明提出的药盒配方在国际上没有相同的报道。Beneficial effect of the present invention: we think, cause the cause of the unstable labeling rate of the medicine box of Choi S design to be: (1) do not contain pH buffering agent in the system, make pH value can not be maintained at fixed value in the labeling process; (2) Too little TRODAT-1 was used. Due to the unstable chemical properties of TRODAT-1, some or even all of TRODAT-1 may decompose and deteriorate during the process of preparation, transportation and storage. The present invention has been specifically improved: (1) a pH buffering agent is added in the formula, so that the pH value can be maintained at a stable value during the labeling process, and the labeling rate is stable and greater than 90%, which is very beneficial to clinical medication; (2) improves When the amount of TRODAT-1 in the kit is reduced, when a small amount of TRODAT-1 deteriorates, there is still a sufficient amount of TRODAT-1 in the kit to ensure that the labeling rate is greater than 90%. The medicine box also has the characteristics that the marking process is simple and can be popularized and applied conveniently. The medicine box formula that the present invention proposes does not have identical report in the world.

附图说明Description of drawings

图1.TRODAT-1的含量对标记率的影响。Figure 1. The effect of the content of TRODAT-1 on the labeling rate.

图2.亚锡盐含量对标记率的影响。Figure 2. Effect of stannous salt content on labeling efficiency.

图3.乙二胺四乙酸盐(EDTA)含量对标记率的影响。Figure 3. Effect of ethylenediaminetetraacetic acid salt (EDTA) content on labeling efficiency.

图4.葡庚糖酸盐(GH)含量对标记率的影响。Figure 4. Effect of glucoheptonate (GH) content on labeling efficiency.

图5.药盒的pH对标记率的影响。Figure 5. The effect of the pH of the kit on the labeling rate.

具体实施方式Detailed ways

实施例1Example 1

TRODAT-1药盒的制备：Preparation of TRODAT-1 kit:

100瓶TRODAT-1药盒的制备方法如下：称取磷酸二氢钾2mmol，磷酸氢二钠5mmol，溶于88mL蒸馏水中，再加入1.00g的葡庚糖酸钠和100mg的乙二胺四乙酸二钠，加入1.00mg/mL的氯化亚锡稀盐酸(0.05mol/L)溶液7.00mL，最后加入1.00mg/mL的TRODAT-1稀盐酸(0.05mol/L)溶液5.00mL，混匀后无菌过滤，再按1.00mL/瓶分装于10ml西林瓶中，冷冻干燥后即得TRODAT-1药盒。每支药盒中含50.0μg的TRODAT-1、70.0μg的氯化亚锡、1.00mg的乙二胺四乙酸二钠和10.0mg的葡庚糖酸钠，药盒中加入1ml水后pH值为：4.0-8.5。The preparation method of 100 bottles of TRODAT-1 kit is as follows:Weigh 2 mmol of potassium dihydrogen phosphate and 5 mmol of disodium hydrogen phosphate, dissolve them in 88 mL of distilled water, then add 1.00 g of sodium glucoheptonate and 100 mg of ethylenediaminetetraacetic acid Disodium, add 7.00mL of 1.00mg/mL stannous chloride dilute hydrochloric acid (0.05mol/L) solution, and finally add 5.00mL of 1.00mg/mL TRODAT-1 dilute hydrochloric acid (0.05mol/L) solution, mix well Sterile filtration, and then divided into 10ml vials at 1.00mL/bottle, and freeze-dried to obtain the TRODAT-1 kit. Each kit contains 50.0 μg of TRODAT-1, 70.0 μg of stannous chloride, 1.00 mg of disodium EDTA and 10.0 mg of sodium glucoheptonate, and the pH value after adding 1 ml of water to the kit For: 4.0-8.5.

实施例2Example 2

TRODAT-1药盒的标记方法：Labeling method of TRODAT-1 kit:

取TRODAT-1药盒1支，加入新淋洗的Na^99mTcO4溶液1.0mL，置于沸水浴上加热30min，冷却后即可。Take 1 TRODAT-1 kit, add 1.0mL of newly washed Na^99m TcO4 solution, place it on a boiling water bath and heat it for 30min, then cool it down.

实施例3Example 3

标记率的测定方法：Marking rate measurement method:

上行薄层色谱(TLC)法。Upstream thin layer chromatography (TLC) method.

实施例4Example 4

各组份含量在一定范围内变化时对药盒标记率的影响：The influence of the content of each component on the labeling rate of the kit when the content of each component changes within a certain range:

保持其它组份含量按实施例1中的不变，改变药盒中TRODAT-1的含量为1-300μg，按实施例2的方法标记后再按实施例3的方法测定标记率。结果如图1所示，TRODAT-1的含量在20-200μg范围内，药盒标记率达到90％。Keep the content of other components unchanged as in Example 1, change the content of TRODAT-1 in the kit to 1-300 μg, mark according to the method of Example 2, and then measure the labeling rate according to the method of Example 3. The results are shown in Figure 1, the content of TRODAT-1 was in the range of 20-200 μg, and the labeling rate of the kit reached 90%.

实施例5Example 5

各组份含量在一定范围内变化时对药盒标记率的影响：The influence of the content of each component on the labeling rate of the kit when the content of each component changes within a certain range:

保持其它组份含量按实施例1中的不变，药盒中亚锡盐改用氟化亚锡，其含量为0.5-100μg，按实施例2的方法标记后再按实施例3的方法测定标记率。结果如图2所示，亚锡盐的含量在4-100μg范围内，药盒标记率达到90％。Keep the content of other components constant as in Example 1, use stannous fluoride instead of stannous salt in the kit, its content is 0.5-100 μ g, and then measure by the method of Example 3 after marking by the method of Example 2 marking rate. The results are shown in Figure 2, the content of the stannous salt is in the range of 4-100 μg, and the labeling rate of the kit reaches 90%.

实施例6Example 6

各组份含量在一定范围内变化时对药盒标记率的影响：The influence of the content of each component on the labeling rate of the kit when the content of each component changes within a certain range:

保持其它组份含量按实施例1中的不变，药盒中乙二胺四乙酸盐改用乙二胺四乙酸二钾，其含量为0.1-20mg，按实施例2的方法标记后再按实施例3的方法测定标记率。结果如图3所示，乙二胺四乙酸盐的含量在0.5-10mg范围内，药盒标记率达到90％。Keep the content of other components constant as in Example 1, use dipotassium edetate instead of edetate in the kit, and its content is 0.1-20mg, mark it according to the method of example 2 and then The labeling rate was determined according to the method in Example 3. The results are shown in Figure 3, the content of EDTA is in the range of 0.5-10 mg, and the labeling rate of the kit reaches 90%.

实施例7Example 7

各组份含量在一定范围内变化时对药盒标记率的影响：The influence of the content of each component on the labeling rate of the kit when the content of each component changes within a certain range:

保持其它组份含量按实施例1中的不变，药盒中葡庚糖酸盐改用葡庚糖酸钾，其含量为1-20mg，按实施例2的方法标记后再按实施例3的方法测定标记率。结果如图4所示，葡庚糖酸盐的含量在1-20mg范围内，药盒标记率达到90％。Keep the content of other components unchanged as in Example 1, use potassium glucoheptonate in the kit instead of glucoheptonate, and its content is 1-20 mg, mark according to the method of Example 2 and then follow Example 3 method to determine the labeling rate. The results are shown in Figure 4, the content of glucoheptonate is in the range of 1-20 mg, and the labeling rate of the kit reaches 90%.

实施例8Example 8

药盒的pH值对标记率的影响：Effect of the pH value of the kit on the labeling rate:

保持其它组份含量按实施例1中的不变，用pH缓冲剂调节药盒pH值在1-12范围内变化，按实施例2的方法标记后再按实施例3的方法测定标记率。结果图示5所示，pH值在4.0-8.5范围内药盒标记率大于90％。Keep the content of other components constant as in Example 1, adjust the pH value of the kit to vary within the range of 1-12 with a pH buffer, and then measure the labeling rate by the method of Example 3 after marking by the method of Example 2. As shown in Figure 5 of the results, the labeling rate of the kit was greater than 90% within the pH range of 4.0-8.5.

所用pH缓冲剂为磷酸盐、柠檬酸盐、铵盐、醋酸盐、酒石酸盐、碳酸盐、硼酸盐、巴比妥盐、邻苯二钾酸盐中的一种或一种以上的混合物。The pH buffer used is one or more of phosphate, citrate, ammonium salt, acetate, tartrate, carbonate, borate, barbiturate, phthalate mixture.

Claims (6)

1. the medicine box of β-[N, N '-two (2-mercapto ethyl) ethylene diamin(e)] methyl-3 β-(4-chlorphenyl) tropane, be used to carry out technetium [^99mTc] labelling, it is characterized in that consisting of: 2 β-[N, N '-two (2-mercapto ethyl) ethylene diamin(e)] methyl-3 β-(4-chlorphenyl) tropane, tin salt, gluceptate, edetate, pH buffer agent, the content range of each component is in the medicine box:

2 β-[N, N '-two (2-mercapto ethyl) ethylene diamin(e)] methyl

-3 β-(4-chlorphenyl) tropane 20-200 μ g

Tin salt 4-100 μ g

Edetate 0.5-10mg

Gluceptate 1-20mg

It is 4.0-8.5 that the pH buffer agent is regulated the pH scope.

2. medicine box according to claim 1 is characterized in that tin salt is stannous chloride, stannous fluoride, stannous tartrate, Tin dibromide. or stannous citrate.

3. medicine box according to claim 1 is characterized in that edetate is disodium edta or ethylene diamine tetraacetic acid sylvite.

4. medicine box according to claim 1 is characterized in that gluceptate is gluceptate sodium or glucoheptonic acid potassium.

5. medicine box according to claim 1 is characterized in that pH buffer agent in the medicine box is one or more the mixture in phosphate, citrate, ammonium salt, acetate, tartrate, carbonate, borate, Barbiturate, the o-2 potassium acid salt.

6. the compound method of medicine box as claimed in claim 1, it is characterized in that 100 bottle of 2 β-[N, N '-two (2-mercapto ethyl) ethylene diamin(e)] preparation method of methyl-3 β-(4-chlorphenyl) tropane medicine box is as follows, take by weighing pH buffer agent 7mmol, be dissolved in the 70mL distilled water, add the gluceptate of 0.1-2.0g and the edetate of 0.05-1.00g again, the 0.05mol/L dilute hydrochloric acid solution 0.4-10mL that adds the tin salt of 1mg/mL, the 2 β-[N that adds 1mg/mL at last, N '-two (2-mercapto ethyl) ethylene diamin(e)] the 0.05mol/L dilute hydrochloric acid solution 2-20mL of methyl-3 β-(4-chlorphenyl) tropane, adding distilled water to medicinal liquid cumulative volume again is 100ml, be sub-packed in the cillin bottle of 10ml specification by the 1.0mL/ bottle again, promptly get 2 β-[N after the lyophilization, N '-two (2-mercapto ethyl) ethylene diamin(e)] methyl-3 β-(4-chlorphenyl) tropane medicine box, the 2 β-[N that contains 20-200 μ g in every medicine box, N '-two (2-mercapto ethyl) ethylene diamin(e)] methyl-3 β-(4-chlorphenyl) tropane, the tin salt of 4-100 μ g, 0.5-10mg edetate and the gluceptate of 1-20mg, medicine box adds that the pH value scope is 4.0-8.5 behind the 1ml distilled water.

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