WO2021232791A1 - Procédé de préparation de polypeptide actif antitumoral de type tubulysine m efficace - Google Patents

Procédé de préparation de polypeptide actif antitumoral de type tubulysine m efficace Download PDF

Info

Publication number
WO2021232791A1
WO2021232791A1 PCT/CN2020/139772 CN2020139772W WO2021232791A1 WO 2021232791 A1 WO2021232791 A1 WO 2021232791A1 CN 2020139772 W CN2020139772 W CN 2020139772W WO 2021232791 A1 WO2021232791 A1 WO 2021232791A1
Authority
WO
WIPO (PCT)
Prior art keywords
compound
reaction
water
add
ethyl acetate
Prior art date
Application number
PCT/CN2020/139772
Other languages
English (en)
Chinese (zh)
Inventor
吴正治
龙伯华
Original Assignee
深圳市老年医学研究所
吴正治
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 深圳市老年医学研究所, 吴正治 filed Critical 深圳市老年医学研究所
Publication of WO2021232791A1 publication Critical patent/WO2021232791A1/fr

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06139—Dipeptides with the first amino acid being heterocyclic

Definitions

  • the present invention relates to the technical field of medicines, and more specifically, to a preparation method of Tubulysin M, a polypeptide with high anti-tumor activity.
  • Tubulysin A-Z is a family of compounds, as shown in Table 1, they are all natural products.
  • the active peptide Tubulysin M is obtained by chemists through structural modification, that is, the sensitive acetal is replaced with a stable methyl group. Studies have found that the active peptide Tubulysin M shows strong cytotoxic activity against human cancer cells, with IC50 values in the low nanomolar range, as shown in Table 2 (Reference J.Am.Chem.Soc., 2016, 138(5) ):1698-1708).
  • the active peptide Tubulysin M is a cytotoxic activity of tubulin, which inhibits tubulin polymerization and leads to cell cycle arrest and apoptosis.
  • the active peptide Tubulysin M also has the advantages of inhibiting angiogenesis and resisting multi-drug resistance. In terms of water solubility, the active peptide Tubulysin M has obvious advantages over other anti-cancer drugs. It shows super anti-cancer activity against specific cancer cells, making it one of the most beloved target molecules in the development of new anti-cancer drugs. one.
  • the present invention provides a method for preparing the active polypeptide Tubulysin M.
  • the preparation method adopts an innovative route and optimizes the total synthesis process of the active polypeptide Tubulysin M.
  • the overall yield is high and the stereoselectivity is high. good.
  • a preparation method of Tubulysin M, a polypeptide with high anti-tumor activity comprising:
  • Step S1 providing compound 2;
  • Step S2 preparing active polypeptide Tubulysin M using compound 2 as a key raw material
  • step S2 adopts the following synthetic route:
  • the third intermediate is dissolved in dichloromethane, trifluoroacetic acid is added, the reaction is stirred at room temperature, concentrated under reduced pressure, and dried to obtain the fourth intermediate;
  • the molar ratio of compound 2: compound 3: HATU: diisopropylethylamine 1:1.1-1.5:2-3:4-6, preferably 1: 1.2:3:6.
  • the volume ratio of petroleum ether: ethyl acetate in the mixture of petroleum ether and ethyl acetate 4:1.
  • the solvent used in the extraction is dichloromethane.
  • the molar ratio of compound 4:2,6-lutidine:tert-butyldimethylsilyl trifluoromethanesulfonate 1:3-5:1.5 -2, preferably 1:3:1.5.
  • 2,6-lutidine and tert-butyldimethylsilyl trifluoromethanesulfonate are added under cooling in an ice water bath, and after 20 to 30 minutes, the temperature is raised to room temperature and the reaction is stirred for 3 to 3 minutes. 4h, more preferably 30min, raise to room temperature and stir for 4h.
  • the dilute hydrochloric acid is 1M dilute hydrochloric acid.
  • the volume ratio of petroleum ether: ethyl acetate in the mixture of petroleum ether and ethyl acetate 8:1.
  • the molar ratio of compound 5: sodium hydride: methyl iodide 1:3-5:1.5-2, preferably 1:5:1.5.
  • the solvent used in the extraction is ethyl acetate.
  • the volume ratio of petroleum ether: ethyl acetate in the mixture of petroleum ether and ethyl acetate 10:1.
  • the molar ratio of compound 6: sodium hydroxide: compound 7: HATU: diisopropylethylamine 1:10-15:1.1-1.5:2-3: 4-6, preferably 1:10: 1.2:3:6.
  • step (4) solid sodium hydroxide is added under cooling in an ice-water bath, and the temperature is raised to room temperature for 20-30 minutes, and the reaction is stirred for 2 to 3 hours, and more preferably, the temperature is raised to room temperature for 30 minutes and the reaction is stirred for 2 hours.
  • the volume ratio of tetrahydrofuran:water in the tetrahydrofuran/water mixed solvent 1:1.
  • the volume ratio of petroleum ether: ethyl acetate in the mixture of petroleum ether and ethyl acetate 8:1.
  • the molar ratio of compound 8: triphenylphosphine: compound 9: HATU: diisopropylethylamine 1:10-15:1.1-1.5:2-3 : 4-6, preferably 1:10:1.2:3:6;
  • the volume ratio of tetrahydrofuran:water in the tetrahydrofuran/water mixed solvent 20:1.
  • the volume ratio of petroleum ether: ethyl acetate in the mixture of petroleum ether and ethyl acetate 4:1.
  • the molar ratio of compound 10: ammonium fluoride: sodium hydroxide: acetic anhydride: trifluoroacetic acid: formaldehyde: sodium cyanoborohydride 1:50-100:10 -15:20-50:20-30:10-20:10-20, preferably 1:100:10:30:20:20:20.
  • the volume ratio of tetrahydrofuran:water in the tetrahydrofuran/water mixed solvent 1:1.
  • the volume ratio of acetonitrile:methanol in the mixed solvent of acetonitrile and methanol 1:1.
  • the mass fraction of the aqueous formaldehyde solution is 37%.
  • step S1 compound 2 adopts the following synthetic route:
  • Step 11 Dissolve the starting material L-valinol 2-1 in a tetrahydrofuran/water mixed solvent, add solid sodium bicarbonate and benzyl chloroformate, and react overnight at room temperature to obtain compound 2-2;
  • Step 12 Dissolve compound 2-2 obtained in step 1 in acetonitrile, add 2-iodoylbenzoic acid, and react with heating under reflux to obtain intermediate aldehyde 2-3;
  • Step 13 Dissolve the compound 2-5 in a mixed solvent of tetrahydrofuran/water, add solid sodium hydroxide, and react with heating under reflux to obtain compound 2-6;
  • Step 14 Dissolve the compound 2-8 in tetrahydrofuran, add concentrated hydrochloric acid, and react at room temperature to obtain compound 2-9;
  • Step 15 Under the protection of inert gas, dissolve the compound 2-9 in anhydrous tetrahydrofuran, add (S)-Me-CBS and borane dimethyl sulfide complex under cooling in an ice water bath, and warm to room temperature After reaction, compound 2 is obtained.
  • the molar ratio of L-valinol 2-1: sodium bicarbonate: benzyl chloroformate is 1:3-5:1-1.02, preferably 1:3:1 .
  • the reaction time in step 11 is 10-15h, more preferably 12h.
  • it is concentrated under reduced pressure, extracted with ethyl acetate, combined the organic phases, washed with saturated brine, separated and collected the organic phase, and then Anhydrous sodium sulfate was added to the organic phase to dry, filtered, concentrated and drained to obtain compound 2-2.
  • the molar ratio of compound 2-2:2-iodoylbenzoic acid:Wittig reagent 2-4 is 1:2-3:1.2-1.5, preferably 1:2:1.5.
  • the present invention has the following beneficial effects:
  • the preparation method of the present invention adopts an innovative route, optimizes the total synthesis process of the active polypeptide Tubulysin M, has a high total yield and good stereoselectivity.
  • the synthetic route adopted by the present invention utilizes renewable resources, is recyclable, the reagents used are less toxic, and the reaction has little environmental pollution after treatment, and meets the green chemical standards.
  • the preparation method of the present invention has convenient experimental operation, mild reaction conditions, simple separation and purification, can be used for large-scale preparation, and is convenient for industrialization.
  • HATU in the present invention 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate; DIPEA: diisopropylethylamine; DCM: two Methyl chloride; TBSOTf: tert-butyl dimethylsilyl trifluoromethanesulfonate.
  • DIPEA diisopropylethylamine
  • DCM two Methyl chloride
  • TBSOTf tert-butyl dimethylsilyl trifluoromethanesulfonate.
  • the English abbreviations of other chemical reagents used are all conventional meanings in the art for understanding.
  • the present invention provides a preparation method of Tubulysin M, a polypeptide with high anti-tumor activity, and the preparation method includes:
  • Step S1 providing compound 2;
  • Step S2 preparing active polypeptide Tubulysin M using compound 2 as a key raw material
  • step S1 The specific method for providing compound 2 in step S1 can follow the preparation method described in Chinese patent application 202010336478.7, of course, the compound 2 can also be obtained in other ways.
  • Step S2 specifically adopts the following synthetic route:
  • the synthesized compound 4 was tested by 1 HNMR, 13 CNMR, and HRMS.
  • Dissolve compound 5 (3.0g, 5.86mmol) in anhydrous DMF (200mL), add 60% sodium hydride solid (0.6g, 15mmol) under ice-water bath cooling, add iodomethane (0.62mL, 10mmol) after 30 minutes, slowly increase
  • the reaction was stirred at room temperature for 2 hours, cooled in an ice-water bath, quenched by adding saturated aqueous ammonium chloride solution (500 mL), extracted three times with ethyl acetate (300 mL), and the combined organic phase was washed with water (200 mL), washed with saturated brine (200 mL), and separated.
  • the organic phase was dried with anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude product.
  • the preparation method of compound 7 can refer to patent application 202010393017.3.
  • the total yield of compound 10 in two steps is 70%.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

L'invention concerne un procédé de préparation d'un polypeptide actif antitumoral de type tubulysine M efficace. Ce procédé comprend les étapes consistant à : S1, fournir le composé 2; et S2, préparer le polypeptide actif de type tubulysine M en prenant le composé 2 en tant que matière première clé. Dans le procédé de préparation, une voie innovante est utilisée, le processus de synthèse totale du polypeptide actif de type tubulysine M est optimisé, le rendement total est relativement élevé, et la stéréosélectivité est bonne.
PCT/CN2020/139772 2020-05-22 2020-12-26 Procédé de préparation de polypeptide actif antitumoral de type tubulysine m efficace WO2021232791A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN202010438990.2A CN111690036A (zh) 2020-05-22 2020-05-22 一种高效抗肿瘤活性多肽Tubulysin M的制备方法
CN202010438990.2 2020-05-22

Publications (1)

Publication Number Publication Date
WO2021232791A1 true WO2021232791A1 (fr) 2021-11-25

Family

ID=72477211

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2020/139772 WO2021232791A1 (fr) 2020-05-22 2020-12-26 Procédé de préparation de polypeptide actif antitumoral de type tubulysine m efficace

Country Status (2)

Country Link
CN (1) CN111690036A (fr)
WO (1) WO2021232791A1 (fr)

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114478529A (zh) * 2022-03-01 2022-05-13 无锡捷化医药科技有限公司 一种cdk抑制剂及其制备方法
CN114671796A (zh) * 2022-04-22 2022-06-28 宁波工程学院 一种光催化合成n-烷基邻苯二甲酰亚胺的方法
CN114957284A (zh) * 2022-06-07 2022-08-30 中国科学院生态环境研究中心 天然产物Lycibarbarine的高效合成方法及应用
CN115536529A (zh) * 2022-09-26 2022-12-30 浦拉司科技（上海）有限责任公司 一种3,5-二（2-氰基-异丙基）-甲苯的合成方法
CN116143758A (zh) * 2022-12-21 2023-05-23 天津科技大学 一类氮杂黄酮类靶向蛋白嵌合体及其在制备抗肿瘤药物中的应用
CN116199680A (zh) * 2022-12-21 2023-06-02 大连理工大学 含gpx4蛋白共价基团的内过氧类化合物及其制备方法与应用

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111690036A (zh) * 2020-05-22 2020-09-22 深圳市老年医学研究所 一种高效抗肿瘤活性多肽Tubulysin M的制备方法

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102695714A (zh) * 2009-11-12 2012-09-26 R&D生技药品有限责任公司 微管蛋白抑制剂
WO2017031209A1 (fr) * 2015-08-18 2017-02-23 Endocyte, Inc. Analogues de tubulysine et procédés associés
CN109678929A (zh) * 2019-01-17 2019-04-26 深圳市老年医学研究所 一种生物活性肽N14-Desacetoxytubulysin H的制备方法
CN109912683A (zh) * 2017-12-13 2019-06-21 杭州多禧生物科技有限公司 一种细胞毒素分子、偶联物及其制备方法和应用
CN111647040A (zh) * 2020-05-11 2020-09-11 深圳市老年医学研究所 一种天然活性多肽Tubulysin U的制备方法
CN111690036A (zh) * 2020-05-22 2020-09-22 深圳市老年医学研究所 一种高效抗肿瘤活性多肽Tubulysin M的制备方法

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102695714A (zh) * 2009-11-12 2012-09-26 R&D生技药品有限责任公司 微管蛋白抑制剂
WO2017031209A1 (fr) * 2015-08-18 2017-02-23 Endocyte, Inc. Analogues de tubulysine et procédés associés
CN109912683A (zh) * 2017-12-13 2019-06-21 杭州多禧生物科技有限公司 一种细胞毒素分子、偶联物及其制备方法和应用
CN109678929A (zh) * 2019-01-17 2019-04-26 深圳市老年医学研究所 一种生物活性肽N14-Desacetoxytubulysin H的制备方法
CN111647040A (zh) * 2020-05-11 2020-09-11 深圳市老年医学研究所 一种天然活性多肽Tubulysin U的制备方法
CN111690036A (zh) * 2020-05-22 2020-09-22 深圳市老年医学研究所 一种高效抗肿瘤活性多肽Tubulysin M的制备方法

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
LONG, B. ET AL.: ""Total synthesis of tubulysin U and N14-desacetoxy tubulysin H"", ORGANIC & BIOMOLECULAR CHEMISTRY, vol. 18, 3 July 2020 (2020-07-03) *
PETER WIPF, TAKESHI TAKADA, MICHAEL J. RISHEL: "Synthesis of the Tubuvaline-Tubuphenylalanine (Tuv-Tup) Fragment of Tubulysin", ORGANIC LETTERS, AMERICAN CHEMICAL SOCIETY, US, vol. 6, no. 22, 1 October 2004 (2004-10-01), US , pages 4057 - 4060, XP055532591, ISSN: 1523-7060, DOI: 10.1021/ol048252i *

Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114478529A (zh) * 2022-03-01 2022-05-13 无锡捷化医药科技有限公司 一种cdk抑制剂及其制备方法
CN114671796A (zh) * 2022-04-22 2022-06-28 宁波工程学院 一种光催化合成n-烷基邻苯二甲酰亚胺的方法
CN114671796B (zh) * 2022-04-22 2024-06-11 宁波工程学院 一种光催化合成n-烷基邻苯二甲酰亚胺的方法
CN114957284A (zh) * 2022-06-07 2022-08-30 中国科学院生态环境研究中心 天然产物Lycibarbarine的高效合成方法及应用
CN114957284B (zh) * 2022-06-07 2023-06-09 中国科学院生态环境研究中心 天然产物Lycibarbarine的高效合成方法及应用
CN115536529A (zh) * 2022-09-26 2022-12-30 浦拉司科技（上海）有限责任公司 一种3,5-二（2-氰基-异丙基）-甲苯的合成方法
CN116143758A (zh) * 2022-12-21 2023-05-23 天津科技大学 一类氮杂黄酮类靶向蛋白嵌合体及其在制备抗肿瘤药物中的应用
CN116199680A (zh) * 2022-12-21 2023-06-02 大连理工大学 含gpx4蛋白共价基团的内过氧类化合物及其制备方法与应用
CN116199680B (zh) * 2022-12-21 2024-04-02 大连理工大学 含gpx4蛋白共价基团的内过氧类化合物及其制备方法与应用
CN116143758B (zh) * 2022-12-21 2024-04-23 天津科技大学 一类氮杂黄酮类靶向蛋白嵌合体及其在制备抗肿瘤药物中的应用

Also Published As

Publication number Publication date
CN111690036A (zh) 2020-09-22

Similar Documents

Publication Publication Date Title
WO2021232791A1 (fr) Procédé de préparation de polypeptide actif antitumoral de type tubulysine m efficace
WO2021227512A1 (fr) Procédé de préparation du polypeptide naturel actif tubulysine u
CN101967145B (zh) 一种抗血栓药物阿匹沙班的制备方法
CN111116676B (zh) 一种具有蝶烯结构的n-杂环卡宾钯配合物及其应用
CN109734738B (zh) 一种能快速检测亚硫酸氢根离子的荧光探针及其制备方法与应用
CN112538079B (zh) 一种香豆素类衍生物及其合成方法与应用
CN114395009B (zh) 一种高纯度胆固醇的合成方法
CN102659851B (zh) 多金属氧酸盐-倍半硅氧烷杂化化合物及制备方法
CN109096339B (zh) 一种三联吡啶钌配合物的制备及在逆转录酶抑制中的应用
CN104151861B (zh) 一种以菲并咔唑为核心的有机染料及其制备方法，以及有机染料敏化太阳电池
CN110437172A (zh) 一类3,7-二取代吩噁嗪类衍生物及其制备方法
CN111825655B (zh) 一种检测Hg2+用高灵敏性荧光探针及其制备方法和应用
CN101531654B (zh) 一种卢帕他定的制备方法
CN108484641B (zh) 依度沙班对甲苯磺酸盐一水合物的制备方法
CN104311485B (zh) 一种治疗白血病的药物博舒替尼的制备方法
CN110981832A (zh) 一种盐酸罗沙替丁醋酸酯的制备方法
CN104761599B (zh) 一种5,4’‑二羟基黄酮‑7‑o‑d‑葡萄糖醛酸的制备方法
WO2023109273A1 (fr) Procédé de préparation d'islatravir médicament anti-vih
CN111269094B (zh) 2-溴-1,3-二甲氧基苯的制备方法
CN107216271A (zh) 酒石酸匹莫范色林杂质及其制备方法
CN110981715B (zh) 一种蛋白质化学交联剂及其制备方法与应用
CN105111229B (zh) 一种硅噻菌胺的合成方法
CN112442005B (zh) 一种n-磺酰基亚胺香豆素衍生物及其制备方法
CN105399602B (zh) 一种v‑型刚性双齿羧酸配体及其制备方法和应用
CN102731388A (zh) 一种（r）/（s）-6,6’-二羟基-5,5’-双喹啉的制备方法

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 20936378

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 20936378

Country of ref document: EP

Kind code of ref document: A1