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WO2020087794A1 - 一种f-18标记的5-氨基乙酰丙酸衍生物、合成方法及应用 - Google Patents

一种f-18标记的5-氨基乙酰丙酸衍生物、合成方法及应用 Download PDF

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WO2020087794A1
WO2020087794A1 PCT/CN2019/073553 CN2019073553W WO2020087794A1 WO 2020087794 A1 WO2020087794 A1 WO 2020087794A1 CN 2019073553 W CN2019073553 W CN 2019073553W WO 2020087794 A1 WO2020087794 A1 WO 2020087794A1
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reaction
aminolevulinic acid
ala
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labeled
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庄晓青
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庄晓青
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C229/00—Compounds containing amino and carboxyl groups bound to the same carbon skeleton
    • C07C229/02—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton
    • C07C229/04—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated
    • C07C229/22—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated the carbon skeleton being further substituted by oxygen atoms
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
    • A61K51/02—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by the carrier, i.e. characterised by the agent or material covalently linked or complexing the radioactive nucleus
    • A61K51/04—Organic compounds
    • A61K51/0404—Lipids, e.g. triglycerides; Polycationic carriers
    • A61K51/0406—Amines, polyamines, e.g. spermine, spermidine, amino acids, (bis)guanidines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
    • A61K51/02—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by the carrier, i.e. characterised by the agent or material covalently linked or complexing the radioactive nucleus
    • A61K51/04—Organic compounds
    • A61K51/0497—Organic compounds conjugates with a carrier being an organic compounds
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
    • C07B2200/00—Indexing scheme relating to specific properties of organic compounds
    • C07B2200/05—Isotopically modified compounds, e.g. labelled
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00—Technologies relating to chemical industry
    • Y02P20/50—Improvements relating to the production of bulk chemicals
    • Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups

Definitions

  • the embodiments of the present application relate to the technical field of organic compound synthesis, in particular to an F-18-labeled 5-aminolevulinic acid derivative, synthesis method, and application.
  • 5-Aminolevulinic acid is an endogenous precursor substance of heme metabolism in human body, which is catalyzed by glycine and succinyl CoA in 5-ALA synthase in mitochondria Under synthesis. It has no photosensitivity in itself, but it produces protoporphyrin IX (PpIX) with near infrared fluorescence and photosensitivity after being metabolized in the body. The formation process of protoporphyrin IX in normal cells is very slow, and under the action of ferrochelatase (Ferrochelatase) and Fe 2+ ions form heme with no photoactivity.
  • ferrochelatase Ferochelatase
  • protoporphyrin IX in malignant tumor cells is high and accumulated in large amounts, and emits near infrared fluorescence (Near Infrared, NIR).
  • NIR near infrared fluorescence
  • PpIX is a very effective photosensitizer. Under the irradiation of a certain wavelength of light, it undergoes a photochemical reaction to produce singlet oxygen and oxide, causing damage to cell membranes, mitochondria and nucleic acids, thereby causing cancer cells to die and wither. Death, play a therapeutic role. Therefore, 5-ALA is widely used in the diagnosis of various malignant tumors, photodynamic therapy for tumor treatment, and photodynamic diagnosis-oriented tumor resection surgery.
  • radionuclides have very ideal physical and chemical properties, but because the 5-ALA molecule is very small, it can only be labeled with a positron nuclide that does not have a significant impact on its molecular structure.
  • Suzuki et al. First reported that the precursor activated with Schiff base was subjected to 11 C methylation reaction at the 5 position to obtain 11 C-labeled radioactive probe 11 C-MALA. Then they initially evaluated the uptake of 11 C-MALA in various tumor cells and tumor models, and concluded that the uptake of 11 C-MALA is highly correlated with the aggregation of PpIX.
  • the ratio between the uptake of tumor brain tissue and the uptake of normal brain tissue at a 10-minute peak is only about 1.3, which is largely due to the 13 N
  • the half-life is only 9.96 minutes, and the biological half-life of 5-ALA in the body is about 1 hour, so it is difficult to achieve the desired effect.
  • its radioactive half-life should be greater than the half-life of the molecule's biological metabolism in the body, so that molecules in non-target organs have sufficient time for in vivo transport, metabolism, and excretion in vitro, resulting in high targets / Background ratio.
  • the purpose of the present application is to provide an F-18-labeled 5-aminolevulinic acid derivative, a synthesis method and an application to solve the problem of short half-life of radioactive probes in the prior art.
  • an F-18-labeled 5-aminolevulinic acid derivative including 18 F-ALA, whose chemical structure is:
  • n 1-6.
  • an F-18-labeled 5-aminolevulinic acid derivative including 18 FO-ALA, whose chemical structure is:
  • n 2-4.
  • a method for synthesizing an F-18-labeled 5-aminolevulinic acid derivative including:
  • the HBA was added to the dry pyridine, p-toluenesulfonyl chloride was added in portions under ice bath, and then gradually returned to room temperature, stirred overnight, after the reaction, the solution was concentrated, and the column was separated to obtain the precursor THBA;
  • the 18 F - ions from the accelerator were captured by the QMA column, and then eluted with aminopolyether / potassium carbonate to the reaction tube.
  • the acetonitrile was added to repeatedly remove the dry solvent.
  • the precursor THBA was added to the reaction tube and dissolved in dry acetonitrile.
  • the reaction was heated under reflux. After the reaction, the HCl solution was added to heat to remove the protecting group. After dilution with water, it was preliminarily purified with a solid phase extraction column. Finally, the column was separated by high performance liquid chromatography to obtain 18 F-ALA.
  • a method for synthesizing an F-18-labeled 5-aminolevulinic acid derivative including;
  • the HEBA was added to the dry pyridine, p-toluenesulfonyl chloride was added in portions under ice bath, and then gradually returned to room temperature, stirred overnight, after the reaction, the solution was concentrated, and the column was separated to obtain the precursor TEBA;
  • the 18 F - ions from the accelerator were captured by the QMA column, and then eluted with amino polyether / potassium carbonate to the reaction tube.
  • the solvent was repeatedly removed by adding acetonitrile.
  • the precursor TEBA was added to the reaction tube and dissolved in dry acetonitrile.
  • the reaction was heated under reflux.
  • the HCl solution was added to heat to remove the protective group.
  • the solid phase extraction column was used for preliminary purification. Finally, the column was separated by high performance liquid chromatography to obtain 18 FO-ALA.
  • an F-18-labeled 5-aminolevulinic acid derivative is provided for detecting tumors.
  • an F-18-labeled 5-aminolevulinic acid derivative is provided for tumor detection.
  • the embodiments of the present application provide an F-18-labeled 5-aminolevulinic acid derivative, a synthesis method, and an application.
  • the radioactive probe provided in the implementation of this application has a long radioactive half-life, and its stability and fat solubility are obviously higher than that of 5-ALA.
  • F-18-labeled 5-aminolevulinic acid derivatives can form higher concentrations in diseased tissues, while they are lower in normal tissues and less toxic. And because of its better fat solubility and stronger ability to penetrate the blood-brain barrier, it can accurately locate tumor tissues throughout the body and become a broad-spectrum tumor imaging agent.
  • an F-18-labeled 5-aminolevulinic acid derivative including 18 F-ALA, and its chemical structure is:
  • n 1-6.
  • an F-18-labeled 5-aminolevulinic acid derivative including 18 FO-ALA, and its chemical structure is:
  • n 2-4.
  • a method for synthesizing an F-18-labeled 5-aminolevulinic acid derivative including:
  • the HBA was added to the dry pyridine, p-toluenesulfonyl chloride was added in portions under ice bath, and then gradually returned to room temperature, and stirred overnight. After the reaction, the solution was concentrated and separated by the column to obtain the precursor THBA (6- (tosyloxy) hexyl5 -((tert-butoxycarbonyl) amino) -4-oxopentanoate);
  • the 18 F - ions from the accelerator were captured by the QMA column, and then eluted with aminopolyether / potassium carbonate to the reaction tube.
  • the acetonitrile was added to repeatedly remove the dry solvent.
  • the precursor THBA was added to the reaction tube and dissolved in dry acetonitrile.
  • the reaction was heated under reflux. After the reaction, the HCl solution was added to heat to remove the protecting group. After dilution with water, it was preliminarily purified with a solid phase extraction column. Finally, the column was separated by high performance liquid chromatography to obtain 18 F-ALA.
  • the synthetic route of 18 F-ALA is as follows.
  • Boc-5-ALA, dimethylaminopyridine (DMAP), and 1,6-dihexanol are added at a molar ratio of 1: 1: 1 in an ice bath and added with the same molar amount of carbodiimide as Boc-5-ALA.
  • TsCl p-toluenesulfonyl chloride
  • the 18 F - ions from the accelerator were captured by the QMA column, then eluted with K222 / K 2 CO 3 to the reaction tube, and the solvent was removed repeatedly by adding acetonitrile. Then add 5-20 mg of HBA to the reaction tube and dissolve it in 1 mL of dry acetonitrile. Reflux and heat to react. After the reaction, 2M HCl solution was added to remove the protecting group by heating, then diluted with water, and then preliminarily purified with a solid phase extraction column. Finally, the column was separated by HPLC, and finally the radiochemical purity of 18 F-ALA-3 was obtained> 95%.
  • a method for synthesizing an F-18-labeled 5-aminolevulinic acid derivative including;
  • the HEBA was added to the dry pyridine, p-toluenesulfonyl chloride was added in portions under ice bath, and then gradually returned to room temperature, and stirred overnight, after the reaction, the solution was concentrated, and the column was separated to obtain the precursor TEBA (2- (2- (tosyloxy ) ethoxy) ethyl 5-((tert-butoxycarbonyl) amino) -4-oxopentanoate);
  • the 18 F - ions from the accelerator were captured by the QMA column, and then eluted with amino polyether / potassium carbonate to the reaction tube.
  • the solvent was repeatedly removed by adding acetonitrile.
  • the precursor TEBA was added to the reaction tube and dissolved in dry acetonitrile.
  • the reaction was heated under reflux.
  • the HCl solution was added to heat to remove the protective group.
  • the solid phase extraction column was used for preliminary purification. Finally, the column was separated by high performance liquid chromatography to obtain 18 FO-ALA.
  • HEBA p-toluenesulfonyl chloride
  • the 18 F - ions from the accelerator were captured by the QMA column, then eluted with K222 / K 2 CO 3 to the reaction tube, and the solvent was removed repeatedly by adding acetonitrile. Then add 5-20 mg of TEBA to the reaction tube and dissolve it in 1 mL of dry acetonitrile, and heat under reflux to carry out the reaction. After the reaction, 2M HCl solution was added to remove the protecting group by heating, then diluted with water, and then preliminarily purified with a solid phase extraction column. Finally, the column was separated by HPLC, and finally the radiochemical purity of 18 FO-ALA-2 was obtained> 95%.
  • the 5-aminolevulinic acid derivatives include 18 F-ALA for detecting tumors.
  • Nude mice were used to make tumor models, tumor-bearing mice were anesthetized and placed on animal PET / CT, then 0.1-0.5mCi 18 F-ALA solution was injected from the tail vein, and metabolized in the body for 0.5-2 hours for PET / CT imaging. image.
  • AsPC-1 cells were used to construct a tumor model, and 0.1-0.5mCi of 18 F-ALA-3 probe was injected from the tail vein.
  • the ⁇ count of each organ in the tumor mice was detected 10 minutes, 1 hour and 2 hours after injection. Value, that is, the amount of radiopharmaceutical 18F-ALA-3, and the ratio of tumor to muscle, the experimental results are shown in Table 1.
  • the distribution data of 18 F-ALA-3 in tumor mice indicates that 18 F-ALA-3 has a higher uptake in the tumor, and the ratio of tumor to muscle is higher, which can be used as a target for tumor Like agent.
  • the 5-aminolevulinic acid derivatives include 18 FO-ALA for detecting tumors.
  • the embodiments of the present application provide an F-18-labeled 5-aminolevulinic acid derivative, a synthesis method, and an application.
  • the radioactive probe provided in the implementation of this application has a long radioactive half-life, and its stability and fat solubility are obviously higher than that of 5-ALA.
  • F-18-labeled 5-aminolevulinic acid derivatives can form higher concentrations in diseased tissues, but lower concentrations in normal tissues and less toxicity. And because of its better fat solubility and stronger ability to penetrate the blood-brain barrier, it can accurately locate tumor tissues throughout the body and become a broad-spectrum tumor imaging agent.

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Abstract

本申请公开了一种F-18标记的5-氨基乙酰丙酸衍生物、合成方法及应用。本申请实施提供的放射性探针放射性半衰期长，稳定性和脂溶性都明显要高于5-氨基乙酰丙酸。与5-氨基乙酰丙酸相比，F-18标记的5-氨基乙酰丙酸衍生物可以在病变组织内形成更高的浓度，而在正常组织内浓度更低，毒性也更小。并且因为它的脂溶性更好，穿透血脑屏障的能力更强，可以对全身肿瘤组织进行精确的定位，成为广谱型肿瘤显像剂。

Description

一种F-18标记的5-氨基乙酰丙酸衍生物、合成方法及应用 技术领域
本申请实施例涉及有机化合物合成技术领域，特别涉及一种F-18标记的5-氨基乙酰丙酸衍生物、合成方法及应用。
背景技术
5-氨基乙酰丙酸(5-aminolevulinic acid，5-ALA)是人体血红素代谢的内源性前体物质，在线粒体内由甘氨酸和琥珀酰辅酶(Succinyl CoA)在5-ALA合成酶的催化下合成。其本身无光敏性，但它在体内代谢后会产生具有近红外荧光和光敏性的原卟啉Ⅸ(protoporphyrin Ⅸ，PpⅨ)。正常细胞中原卟啉Ⅸ的形成过程非常缓慢，并且在亚铁螯合酶(Ferrochelatase)的作用下和Fe 2+离子形成不具有光活性的血红素。但在恶性肿瘤细胞内原卟啉Ⅸ的合成效率高且大量积聚，并发出近红外荧光(Near Infrared，NIR)。并且，PpⅨ是一种非常有效的光敏剂，它在一定波长光的照射下，发生光化学反应，产生单重态氧和氧化物，引起细胞膜、线粒体和核酸的损伤，从而使癌症细胞坏死、凋亡，起到治疗的作用。因此5-ALA被广泛应用于各种恶性肿瘤的诊断，治疗肿瘤的光动力学疗法以及光动力诊断导向的肿瘤切除手术中。
鉴于5-ALA在肿瘤显像方面的优势：1)小分子化合物；2)光敏剂原卟啉PpⅨ的前体；3)肿瘤和正常组织中代谢高度的差异性，如果可以被放射性核素标记，将可以构建一个具有非常高灵敏度的近红外荧光-放射性核素显像双模式探针，对肿瘤进行靶向性的全身显像。
许多放射性核素都有非常理想的物理、化学性质，但是由于5-ALA的分子很小，因此只能用不会对其分子结构造成较大影响的正电子核素进行标记。Suzuki等人首先报道了用席夫碱活化的前体在5位上进行 11C甲基化反应，得到 11C标记的放射性探针 11C-MALA。然后他们初步评价了 11C-MALA在多种肿瘤细胞和肿瘤模型中摄取，并得出结论， 11C-MALA的摄取与PpⅨ的聚集高度相关。同时，从他们的数据中也能看出，从注射后1分钟到60分钟内， 11C-MALA摄取的肿瘤比肌肉的比值一直在升高，这个数值很有可能在此时间段内并没有到达最高点。但是对于半衰期只有20分钟的核素 11C来说，注射后1小时进行检测已经是极限。Pippin等通过用 13N进行标记，完全不改变5-ALA的结构合成了 13N-5-ALA，反应速度快且效率高，并且用 PET可在肿瘤鼠模型中观测到肿瘤内的聚集。但是从他们的数据可以看出，在原位胶质瘤模型中，肿瘤脑组织中的摄取与正常脑组织的摄取在10分钟峰值的时比值只有1.3左右，这很大原因是因为 13N的半衰期只有9.96分钟，而5-ALA在体内的生物半衰期约为1小时，因此很难达到理想的效果。对于一个理想的放射性探针来说，它的放射性半衰期应该大于该分子在体内的生物代谢半衰期，这样才能使在非靶器官的分子有足够时间进行体内运输、代谢以及体外排出，得到高的靶/本底的比值。
发明内容
本申请的目的在于提供一种F-18标记的5-氨基乙酰丙酸衍生物、合成方法及应用，以解决现有技术中放射性探针半衰期短的问题。
第一方面，根据本申请的实施例，提供了一种F-18标记的5-氨基乙酰丙酸衍生物，包括 18F-ALA，其化学结构为：
Figure PCTCN2019073553-appb-000001
其中，n＝1-6。
第二方面，根据本申请的实施例，提供了一种F-18标记的5-氨基乙酰丙酸衍生物，包括 18F-O-ALA，其化学结构为：
Figure PCTCN2019073553-appb-000002
其中，n＝2-4。
第三方面，根据本申请的实施例，提供了一种F-18标记的5-氨基乙酰丙酸衍生物合成方法，包括：
将5-氨基乙酰丙酸溶于水中，调节pH值到8.5，滴加溶于二氧六环的二碳酸二叔丁酯，室温搅拌18h，反应后去除多余的二碳酸二叔丁酯，将水溶液调至酸性，用乙酸乙酯萃取得到5-N-叔丁氧羰基氨基乙酰丙酸；
将所述5-N-叔丁氧羰基氨基乙酰丙酸，二甲氨基吡啶和二醇在冰浴下搅拌，加入碳二亚胺，冰浴搅拌2h，室温继续搅拌48h，反应后旋干溶剂，加入水，用乙酸乙酯进行萃取，收集有机层后用10％的NaHCO 3溶液清洗有机相，最后进行 柱分离，得到淡黄色粉末状的HBA，所述二醇包括二乙醇，二丁醇或二己醇；
在干燥吡啶中加入所述HBA，在冰浴下分批加入对甲苯磺酰氯，然后逐渐恢复到室温，搅拌过夜，反应后溶液浓缩后柱分离，得到前体THBA；
加速器传出来的 18F -离子被QMA柱捕获，然后用氨基聚醚/碳酸钾洗脱至反应管，加乙腈反复除干溶剂，在反应管中加入所述前体THBA溶于干燥乙腈中，回流加热进行反应，反应后加入HCl溶液加热脱除保护基团，加水稀释后用固相萃取柱初步纯化，最后用高效液相色谱法进行柱分离，得到 18F-ALA。
第四方面，根据本申请的实施例，提供了一种F-18标记的5-氨基乙酰丙酸衍生物合成方法，包括；
将5-氨基乙酰丙酸溶于水中，调节pH值到8.5，滴加溶于二氧六环的二碳酸二叔丁酯，室温搅拌18h，反应后去除多余的二碳酸二叔丁酯，将水溶液调至酸性，用乙酸乙酯萃取得到5-N-叔丁氧羰基氨基乙酰丙酸；
将所述5-N-叔丁氧羰基氨基乙酰丙酸，二甲氨基吡啶，以及，二乙二醇，三乙二醇和四乙二醇中的一种在冰浴下搅拌，加入碳二亚胺，冰浴搅拌2h，室温继续搅拌48h，反应后旋干溶剂，加入水，用乙酸乙酯进行萃取，收集有机层后用10％的NaHCO 3溶液清洗有机相，最后进行柱分离，得到淡黄色粉末状的HEBA；
在干燥吡啶中加入所述HEBA，在冰浴下分批加入对甲苯磺酰氯，然后逐渐恢复到室温，搅拌过夜，反应后溶液浓缩后柱分离，得到前体TEBA；
加速器传出来的 18F -离子被QMA柱捕获，然后用氨基聚醚/碳酸钾洗脱至反应管，加乙腈反复除干溶剂，在反应管中加入所述前体TEBA溶于干燥乙腈中，回流加热进行反应，反应后加入HCl溶液加热脱除保护基团，加水稀释后用固相萃取柱初步纯化，最后用高效液相色谱法进行柱分离，得到 18F-O-ALA。
第五方面，根据本申请的实施例，提供了F-18标记的5-氨基乙酰丙酸衍生物应用，用于检测肿瘤。
第六方面，根据本申请的实施例，提供了F-18标记的5-氨基乙酰丙酸衍生物应用，用于检测肿瘤。
由以上技术方案可知，本申请实施例提供一种F-18标记的5-氨基乙酰丙酸衍生物、合成方法及应用。本申请实施提供的放射性探针放射性半衰期长，稳定性和脂溶性都明显要高于5-ALA。与ALA相比，F-18标记的5-氨基乙酰丙酸衍生物 可以在病变组织内形成更高的浓度，而在正常组织内浓度更低，毒性也更小。并且因为它的脂溶性更好，穿透血脑屏障的能力更强，可以对全身肿瘤组织进行精确的定位，成为广谱型肿瘤显像剂。
具体实施方式
根据本申请的实施例，提供了一种F-18标记的5-氨基乙酰丙酸衍生物，包括 18F-ALA，其化学结构为：
Figure PCTCN2019073553-appb-000003
其中，n＝1-6。
当n＝1时， 18F-ALA-1的化学结构为：
Figure PCTCN2019073553-appb-000004
18F-ALA-1的化合物鉴定数据：
质谱(ES+)m/z：177.08
氢谱 1H NMR(CDCl 3):δ1.52(t,2H),δ2.74(t,2H),δ2.84(t,2H),δ3.81(t,2H),δ4.28(t,2H),δ4.37(t,2H)。
当n＝2时， 18F-ALA-2的化学结构为：
Figure PCTCN2019073553-appb-000005
18F-ALA-2的化合物鉴定数据：
质谱(ES+)m/z：205.11
氢谱 1H NMR(CDCl 3):δ1.49(m,2H),δ1.53(t,2H),δ1.62(m,2H),δ2.74(t,2H),δ2.84(t,2H),δ3.81(t,2H),δ4.09(t,2H),δ4.13(t,2H)。
当n＝3时， 18F-ALA-3的化学结构为：
Figure PCTCN2019073553-appb-000006
18F-ALA-3的化合物鉴定数据：
质谱(ES+)m/z：233.14
氢谱 1H NMR(CDCl 3):δ1.43(m,2H)，δ1.49(m,2H)，δ1.53(t,2H)，δ1.62(m,2H)，δ2.74(t,2H)，δ2.84(t,2H)，δ3.84(t,2H)，δ4.13(t,2H)。
当n＝4时， 18F-ALA-4的化学结构为：
Figure PCTCN2019073553-appb-000007
18F-ALA-4的化合物鉴定数据：
质谱(ES+)m/z：261.17
氢谱 1H NMR(CDCl 3):δ1.29(m,6H),δ1.43(m,2H),δ1.49(m,2H),δ1.53(t,2H),δ1.62(m,2H),δ2.74(t,2H),δ2.84(t,2H),δ3.81(t,2H),δ4.09(t,2H),δ4.13(t,2H)。
当n＝5时， 18F-ALA-5的化学结构为：
Figure PCTCN2019073553-appb-000008
18F-ALA-5的化合物鉴定数据：
质谱(ES+)m/z：289.21
氢谱 1H NMR(CDCl 3):δ1.29(m,10H),δ1.43(m,2H),δ1.49(m,2H),δ1.53(t,2H),δ1.62(m,2H),δ2.74(t,2H),δ2.84(t,2H),δ3.81(t,2H),δ4.09(t,2H),δ4.13(t,2H)。
当n＝6时， 18F-ALA-6的化学结构为：
Figure PCTCN2019073553-appb-000009
18F-ALA-6的化合物鉴定数据：
质谱(ES+)m/z：317.24
氢谱 1H NMR(CDCl 3):δ1.26(m,4H),δ1.29(m,10H),δ1.43(m,2H),δ1.49(m,2H),δ1.53(t,2H),δ1.62(m,2H),δ2.74(t,2H),δ2.84(t,2H),δ3.81(t,2H),δ4.09(t,2H),δ4.13(t,2H)。
根据本申请的实施例，提供了一种F-18标记的5-氨基乙酰丙酸衍生物，包括 18F-O-ALA，其化学结构为：
Figure PCTCN2019073553-appb-000010
其中，n＝2-4。
当n＝2时， 18F-O-ALA-2的化学结构为：
Figure PCTCN2019073553-appb-000011
18F-O-ALA-2的化合物鉴定数据：
质谱(ES+)m/z：220.11
氢谱 1H NMR(CDCl 3):δ1.53(t,2H),δ2.74(t,2H),δ2.84(t,2H),δ3.57(t,2H),δ3.65(t,2H),δ3.81(t,2H),δ4.20(t,2H),δ4.26(t,2H)。
当n＝3时， 18F-O-ALA-3的化学结构为：
Figure PCTCN2019073553-appb-000012
18F-O-ALA-3的化合物鉴定数据：
质谱(ES+)m/z：264.14
氢谱 1H NMR(CDCl 3):δ1.53(t,2H),δ2.74(t,2H),δ2.84(t,2H),δ3.54(m,4H),δ3.57(t,2H),δ3.65(t,2H),δ3.81(t,2H),δ4.20(t,2H),δ4.26(t,2H)。
当n＝4时， 18F-O-ALA-4的化学结构为：
Figure PCTCN2019073553-appb-000013
18F-O-ALA-4的化合物鉴定数据：
质谱(ES+)m/z：308.16
氢谱 1H NMR(CDCl 3):δ1.53(t,2H),δ2.74(t,2H),δ2.84(t,2H),δ3.54(m,8H),δ3.57(t,2H),δ3.65(t,2H),δ3.81(t,2H),δ4.20(t,2H),δ4.26(t,2H)。
根据本申请的实施例，提供了一种F-18标记的5-氨基乙酰丙酸衍生物合成方法，包括：
将5-氨基乙酰丙酸溶于水中，调节pH值到8.5，滴加溶于二氧六环的二碳酸二叔丁酯，室温搅拌18h，反应后去除多余的二碳酸二叔丁酯，将水溶液调至酸性，用乙酸乙酯萃取得到5-N-叔丁氧羰基氨基乙酰丙酸；
将所述5-N-叔丁氧羰基氨基乙酰丙酸，二甲氨基吡啶和二醇在冰浴下搅拌，加入碳二亚胺，冰浴搅拌2h，室温继续搅拌48h，反应后旋干溶剂，加入水，用乙酸乙酯进行萃取，收集有机层后用10％的NaHCO 3溶液清洗有机相，最后进行柱分离，得到淡黄色粉末状的HBA(6-hydroxyhexyl5-((tert-butoxycarbonyl)amino)-4-oxopentanoate)，所述二醇包括二乙醇，二丁醇或二己醇；
在干燥吡啶中加入所述HBA，在冰浴下分批加入对甲苯磺酰氯，然后逐渐恢复到室温，搅拌过夜，反应后溶液浓缩后柱分离，得到前体THBA(6-(tosyloxy)hexyl 5-((tert-butoxycarbonyl)amino)-4-oxopentanoate)；
加速器传出来的 18F -离子被QMA柱捕获，然后用氨基聚醚/碳酸钾洗脱至反应管，加乙腈反复除干溶剂，在反应管中加入所述前体THBA溶于干燥乙腈中，回流加热进行反应，反应后加入HCl溶液加热脱除保护基团，加水稀释后用固相萃取柱初步纯化，最后用高效液相色谱法进行柱分离，得到 18F-ALA。
18F-ALA合成路线如下。
Figure PCTCN2019073553-appb-000014
以 18F-ALA-3为例，合成方法如下：
①前体化合物THBA的合成。
A.Boc-5-ALA(5-((tert-butoxycarbonyl)amino)-4-oxopentanoic acid)的合成。
将5-ALA溶于5mL水中，并且用1N的NaOH调节pH值到8.5，然后滴加溶于5mL二氧六环的二碳酸二叔丁酯((Boc) 2O)(相对于5-ALA的1-2倍摩尔量)，室温下搅拌18h。反应后多余的(Boc) 2O用乙醚萃取去除，水溶液用1N的HCl溶液调至酸性，然后用乙酸乙酯萃取得到Boc-5-ALA，减压蒸馏旋干后得到无色油状物。
B.HBA的合成。
Boc-5-ALA，二甲氨基吡啶(DMAP)，和1,6-二己醇按照1:1:1的摩尔比例在冰浴下搅拌加入与Boc-5-ALA同摩尔量的碳二亚胺，然后在冰浴下搅拌2h，之后在室温下继续搅拌48h。反应后旋干溶剂，加入水，然后用乙酸乙酯进行萃取，收集有机层后用10％的NaHCO 3溶液清洗有机相。最后进行柱分离(MeOH:CH 2Cl 2＝40:1)得到淡黄色粉末状HBA。
C.前体THBA的合成。
在干燥的5mL吡啶中加入HBA，在冰浴下分批加入2-3倍摩尔量的对甲苯磺酰氯(TsCl)，然后逐渐恢复到室温，搅拌过夜。反应后溶液浓缩后柱分离。
②放射性探针 18F-ALA-3的标记合成。
加速器传出来的 18F -离子被QMA柱捕获，然后用K222/K 2CO 3洗脱至反应管，加乙腈反复除干溶剂。然后在反应管中加入5-20mg的HBA溶于1mL干燥乙腈中，回流加热进行反应。反应后加入2M HCl溶液加热脱除保护基团，然后加水稀释后用固相萃取柱初步纯化，最后用HPLC进行柱分离，最后得到 18F-ALA-3的放射化学纯度>95％。
18F-ALA-3合成路线如下：
Figure PCTCN2019073553-appb-000015
根据本申请的实施例，提供了一种F-18标记的5-氨基乙酰丙酸衍生物合成方法，包括；
将5-氨基乙酰丙酸溶于水中，调节pH值到8.5，滴加溶于二氧六环的二碳酸二叔丁酯，室温搅拌18h，反应后去除多余的二碳酸二叔丁酯，将水溶液调至酸性，用乙酸乙酯萃取得到5-N-叔丁氧羰基氨基乙酰丙酸；
将所述5-N-叔丁氧羰基氨基乙酰丙酸，二甲氨基吡啶，以及，二乙二醇，三乙二醇和四乙二醇中的一种在冰浴下搅拌，加入碳二亚胺，冰浴搅拌2h，室温继续搅拌48h，反应后旋干溶剂，加入水，用乙酸乙酯进行萃取，收集有机层后用10％的NaHCO 3溶液清洗有机相，最后进行柱分离，得到淡黄色粉末状的HEBA(2-(2-hydroxyethoxy)ethyl 5-((tert-butoxycarbonyl)amino)-4-oxopentanoate)；
在干燥吡啶中加入所述HEBA，在冰浴下分批加入对甲苯磺酰氯，然后逐渐恢复到室温，搅拌过夜，反应后溶液浓缩后柱分离，得到前体TEBA(2-(2-(tosyloxy)ethoxy)ethyl 5-((tert-butoxycarbonyl)amino)-4-oxopentanoate)；
加速器传出来的 18F -离子被QMA柱捕获，然后用氨基聚醚/碳酸钾洗脱至反应管，加乙腈反复除干溶剂，在反应管中加入所述前体TEBA溶于干燥乙腈中，回流加热进行反应，反应后加入HCl溶液加热脱除保护基团，加水稀释后用固相萃取柱初步纯化，最后用高效液相色谱法进行柱分离，得到 18F-O-ALA。
18F-O-ALA合成路线如下。
Figure PCTCN2019073553-appb-000016
以 18F-O-ALA-2为例，合成方法如下：
①前体化合物TEBA的合成。
A.Boc-5-ALA的合成。
将5-ALA溶于5mL水中，并且用1N的NaOH调节pH值到8.5，然后滴加溶于5mL二氧六环的二碳酸二叔丁酯((Boc) 2O)(相对于5-ALA的1-2倍摩尔量)，室温下搅拌18h。反应后多余的(Boc) 2O用乙醚萃取去除，水溶液用1N的HCl溶液调至酸性，然后用乙酸乙酯萃取得到Boc-5-ALA，减压蒸馏旋干后得到无色油状物。
B.HEBA的合成。
Boc-5-ALA，二甲氨基吡啶(DMAP)，和二乙二醇按照1:1:1的摩尔比例在冰浴下搅拌加入同摩尔量的碳二亚胺，然后在冰浴下搅拌2h，之后在室温下继续搅拌48h。反应后旋干溶剂，加入水，然后用乙酸乙酯进行萃取，收集有机层后用10％的NaHCO 3溶液清洗有机相。最后进行柱分离(MeOH:CH 2Cl 2＝40:1)得到淡黄色粉末状HEBA。
C.前体TEBA的合成。
在干燥的5mL吡啶中加入HEBA，在冰浴下分批加入2-3倍摩尔量的对甲苯磺酰氯(TsCl)，然后逐渐恢复到室温，搅拌过夜。反应后溶液浓缩后柱分离。
②放射性探针 18F-O-ALA-2的标记合成。
加速器传出来的 18F -离子被QMA柱捕获，然后用K222/K 2CO 3洗脱至反应管，加乙腈反复除干溶剂。然后在反应管中加入5-20mg的TEBA溶于1mL干燥乙腈中，回流加热进行反应。反应后加入2M HCl溶液加热脱除保护基团，然后加水稀释后用固相萃取柱初步纯化，最后用HPLC进行柱分离，最后得到 18F-O-ALA-2的放射化学纯度>95％。
18F-O-ALA-2合成路线如下。
Figure PCTCN2019073553-appb-000017
根据本申请的实施例，提供了F-18标记的5-氨基乙酰丙酸衍生物应用，所述5-氨基乙酰丙酸衍生物包括 18F-ALA，用于检测肿瘤。
使用裸鼠制作肿瘤模型，将荷瘤鼠麻醉后置于动物PET/CT上，然后从尾静脉注射0.1-0.5mCi的 18F-ALA溶液，在体内代谢0.5-2小时后进行PET/CT显像。
用AsPC-1细胞构建肿瘤模型，从尾静脉注射0.1-0.5mCi的 18F-ALA-3探针，分别在注射后10分钟、1小时和2小时后检测肿瘤鼠中各器官的γ计数的值，即放射性药物18F-ALA-3的量，以及肿瘤和肌肉的比值，实验结果如表1所示。
表1
  10分钟 60分钟 120分钟
血 4.56±1.34 1.14±0.47 0.96±0.11
脑 0.62±0.21 1.02±0.33 1.24±0.18
心脏 2.78±0.41 1.48±0.09 0.96±0.09
肺 4.21±0.62 2.64±0.29 1.26±0.20
肝 9.67±0.63 3.32±0.52 2.37±0.31
脾 13.92±0.58 4.30±0.12 3.20±0.29
胰腺 2.24±0.15 1.64±0.30 1.08±0.09
胃 2.99±0.47 1.24±0.31 1.02±0.08
肠 7.17±0.59 3.98±0.53 2.15±0.76
肾 74.30±7.97 38.64±5.20 18.48±3.24
肌肉 1.02±0.08 0.61±0.09 0.59±0.08
骨 3.02±0.26 1.10±0.47 1.02±0.23
肿瘤 3.98±0.74 5.89±0.65 5.94±0.38
肿瘤/肌肉 3.90±0.71 9.66±0.93 10.07±0.98
由表1可见， 18F-ALA-3在肿瘤鼠的体内分布数据表明 18F-ALA-3在肿瘤内有较高的摄取，并且肿瘤与肌肉的比值较高，可以作为靶向肿瘤的显像剂。
根据本申请的实施例，提供了F-18标记的5-氨基乙酰丙酸衍生物应用，所述5-氨基乙酰丙酸衍生物包括 18F-O-ALA，用于检测肿瘤。
用AsPC-1细胞构建肿瘤模型，从尾静脉注射0.1-0.5mCi的 18F-O-ALA-2探针，分别在注射后10分钟、1小时和2小时后检测肿瘤鼠中各器官的γ计数的值，即放射性药物 18F-O-ALA-2的量，以及肿瘤和肌肉的比值，实验结果如表2所示。
表2
  10分钟 60分钟 120分钟
血 4.02±1.55 1.02±0.35 0.87±0.23
脑 0.53±0.16 0.87±0.26 1.02±0.24
心脏 3.64±0.87 1.76±0.22 0.95±0.07
肺 4.89±1.02 3.42±0.43 1.33±0.27
肝 7.69±1.27 3.04±0.67 1.99±0.43
脾 11.34±2.04 3.89±0.46 2.87±0.22
胰腺 2.36±0.75 1.56±0.45 1.01±0.06
胃 3.45±0.96 1.99±0.57 1.43±0.07
肠 8.78±1.69 4.84±1.24 2.96±0.24
肾 79.9±6.86 49.96±6.32 20.32±2.68
肌肉 1.32±0.07 0.66±0.08 0.52±0.06
骨 3.45±0.31 1.14±0.55 0.99±0.35
肿瘤 4.01±0.53 5.97±1.02 6.26±0.47
肿瘤/肌肉 3.04±0.35 9.05±1.24 12.04±0.66
由表2可见， 18F-O-ALA-2探针在肿瘤鼠的体内分别表明， 18F-O-ALA-2在肿瘤内有较高摄取，肿瘤和肌肉的比值较高，可以作为靶向肿瘤的显像剂。
由以上技术方案可知，本申请实施例提供一种F-18标记的5-氨基乙酰丙酸衍生物、合成方法及应用。本申请实施提供的放射性探针放射性半衰期长，稳定性和脂溶性都明显要高于5-ALA。与ALA相比，F-18标记的5-氨基乙酰丙酸衍生物可以在病变组织内形成更高的浓度，而在正常组织内浓度更低，毒性也更小。并且因为它的脂溶性更好，穿透血脑屏障的能力更强，可以对全身肿瘤组织进行精确的定位，成为广谱型肿瘤显像剂。
本领域技术人员在考虑说明书及实践这里公开的申请后，将容易想到本申请的其它实施方案。本申请旨在涵盖本申请的任何变型、用途或者适应性变化，这些变型、用途或者适应性变化遵循本申请的一般性原理并包括本申请未公开的本技术领域中的公知常识或惯用技术手段。说明书和实施例仅被视为示例性的，本申请的真正范围和精神由下面的权利要求指出。
应当理解的是，本申请并不局限于上面已经描述并在附图中示出的精确结构，并且可以在不脱离其范围进行各种修改和改变。本申请的范围仅由所附的权利要求来限制。

Claims (6)

  1. 一种F-18标记的5-氨基乙酰丙酸衍生物，其特征在于，包括 18F-ALA，其化学结构为：
    Figure PCTCN2019073553-appb-100001
    其中，n＝1-6。
  2. 一种F-18标记的5-氨基乙酰丙酸衍生物，其特征在于，包括 18F-O-ALA，其化学结构为：
    Figure PCTCN2019073553-appb-100002
    其中，n＝2-4。
  3. 一种如权利要求1所述的F-18标记的5-氨基乙酰丙酸衍生物合成方法，其特征在于，包括：
    将5-氨基乙酰丙酸溶于水中，调节pH值到8.5，滴加溶于二氧六环的二碳酸二叔丁酯，室温搅拌18h，反应后去除多余的二碳酸二叔丁酯，将水溶液调至酸性，用乙酸乙酯萃取得到5-N-叔丁氧羰基氨基乙酰丙酸；
    将所述5-N-叔丁氧羰基氨基乙酰丙酸，二甲氨基吡啶和二醇在冰浴下搅拌，加入碳二亚胺，冰浴搅拌2h，室温继续搅拌48h，反应后旋干溶剂，加入水，用乙酸乙酯进行萃取，收集有机层后用10％的NaHCO 3溶液清洗有机相，最后进行柱分离，得到淡黄色粉末状的HBA，所述二醇包括二乙醇，二丁醇或二己醇；
    在干燥吡啶中加入所述HBA，在冰浴下分批加入对甲苯磺酰氯，然后逐渐恢复到室温，搅拌过夜，反应后溶液浓缩后柱分离，得到前体THBA；
    加速器传出来的 18F -离子被QMA柱捕获，然后用氨基聚醚/碳酸钾洗脱至反应管，加乙腈反复除干溶剂，在反应管中加入所述前体THBA溶于干燥乙腈中，回流加热进行反应，反应后加入HCl溶液加热脱除保护基团，加水稀释后用固相萃取柱初步纯化，最后用高效液相色谱法进行柱分离，得到 18F-ALA。
  4. 一种如权利要求2所述的F-18标记的5-氨基乙酰丙酸衍生物合成方法， 其特征在于，包括；
    将5-氨基乙酰丙酸溶于水中，调节pH值到8.5，滴加溶于二氧六环的二碳酸二叔丁酯，室温搅拌18h，反应后去除多余的二碳酸二叔丁酯，将水溶液调至酸性，用乙酸乙酯萃取得到5-N-叔丁氧羰基氨基乙酰丙酸；
    将所述5-N-叔丁氧羰基氨基乙酰丙酸，二甲氨基吡啶，以及，二乙二醇，三乙二醇和四乙二醇中的一种在冰浴下搅拌，加入碳二亚胺，冰浴搅拌2h，室温继续搅拌48h，反应后旋干溶剂，加入水，用乙酸乙酯进行萃取，收集有机层后用10％的NaHCO 3溶液清洗有机相，最后进行柱分离，得到淡黄色粉末状的HEBA；
    在干燥吡啶中加入所述HEBA，在冰浴下分批加入对甲苯磺酰氯，然后逐渐恢复到室温，搅拌过夜，反应后溶液浓缩后柱分离，得到前体TEBA；
    加速器传出来的 18F -离子被QMA柱捕获，然后用氨基聚醚/碳酸钾洗脱至反应管，加乙腈反复除干溶剂，在反应管中加入所述前体TEBA溶于干燥乙腈中，回流加热进行反应，反应后加入HCl溶液加热脱除保护基团，加水稀释后用固相萃取柱初步纯化，最后用高效液相色谱法进行柱分离，得到 18F-O-ALA。
  5. 一种如权利要求1所述的F-18标记的5-氨基乙酰丙酸衍生物应用，其特征在于，用于检测肿瘤。
  6. 一种如权利要求2所述的F-18标记的5-氨基乙酰丙酸衍生物应用，其特征在于，用于检测肿瘤。
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