Medicinal Chemistry Research

, Volume 27, Issue 2, pp 583–591 | Cite as

Enzymic synthesis and biological evaluation of injectable glutathione-everolimus

  • Haibo Wang
  • Xiaohe Zheng
  • Lifei Mao
  • Liang Qin
  • Tianmin Zhu
Original Research


An enzymic synthesis of glutathione-everolimus is reported. This process has been optimized and scaled up with high reproducibility and yields, which will facilitate the development of such conjugate. The stability of the conjugate supported that this prodrug can be prepared into lyophilized solid, which is to be reconstituted with 0.9% sodium chloride for injection before intravenous infusion. And the results of species-related drug release experiment displayed that the performance of the conjugate in human plasma, rat and monkey was similar. Moreover, the in vivo efficacy of glutathione-everolimus in the treatment of renal cell carcinoma was investigated in detail. The conjugate was proved to be an effective, safe and well-tolerated injectable prodrug in the treatment of renal cell carcinoma. The results indicated that three times injection with a high dosage in 1 week can achieve much better in vivo efficacy, and no obvious toxic response was observed.


Everolimus Glutathione Renal cell carcinoma Prodrug Drug delivery mTOR 



This work was financially supported by the Key Science and Technology Innovation Team of Zhejiang Province (2013TD10).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no competing interests.

Supplementary material

44_2017_2084_MOESM1_ESM.pdf (1 mb)
Supplementary Information


  1. Alasker A, Meskawi M, Sun M, Ismail S, Hanna N, Hansen J, Tian Z, Bianchi M, Perrotte P, Karakiewica PI (2013) A contemporary update on rates and management of toxicities of targeted therapies for metastatic renal cell carcinoma. Cancer Treat Rev 39:388–401CrossRefPubMedGoogle Scholar
  2. Arai E, Kanai Y (2010) Genetic and epigenetic alterations during renal carcinogenesis. Int J Clin Exp Pathol 4:58–73PubMedPubMedCentralGoogle Scholar
  3. Coiffier B (2013) Clinical efficacy and management of temsirolimus in patients with relapsed or refractory mantle cell lymphoma. Clin Lymphoma Myeloma Leuk 13:351–359CrossRefPubMedGoogle Scholar
  4. Crowe A, Bruelisauer A, Duerr L, Guntz P, Lemaire M (1999) Absorption and intestinal metabolism of SDZ-RAD and rapamycin in rats. Drug Metab Dispos 27:627–632PubMedGoogle Scholar
  5. Eble JN, Sauter G, Epstein JI, Sesterhenn IA (2004) World Health Organization classification of tumors. IARC, LyonGoogle Scholar
  6. Farag SS, Zhang S, Jansak BS, Wang X, Kraut E, Chan K, Dancey JE, Grever MR (2009) Phase II trial of temsirolimus in patients with relapsed or refractory multiple myeloma. Leuk Res 33:1475–1480CrossRefPubMedPubMedCentralGoogle Scholar
  7. Kirchner GI, Meier-Wiedenbach I, Manns MP (2004) Clinical pharmacokinetics of everolimus. Clin Pharmacokinet 43:83–95CrossRefPubMedGoogle Scholar
  8. Kovarik JM, Hartmann S, Figueiredo J, Rordorf C, Golor G, Lison A, Budde K, Neumayer HH (2002) Effect of food on everolimus absorption: quantification in healthy subjects and a confirmatory screening in patients with renal transplants. Pharmacotherapy 22:154–159CrossRefPubMedGoogle Scholar
  9. Mackenzie MJ, Rini BI, Elson P, Schwandt A, Wood L, Trinkhaus M, Bjarnason G, Knox J (2011) Temsirolimus in VEGF-refractory metastatic renal cell carcinoma. Ann Oncol 22:145–148CrossRefPubMedGoogle Scholar
  10. Motzer RJ, Escudier B, Oudard S, Hutson TE, Porta C, Bracarda S, Grünwald V, Thompson JA, Figlin RA, Hollaender N, Kay A, Ravaud A (2010) Phase 3 trial of everolimus for metastatic renal cell carcinoma: final results and analysis of prognostic factors. Cancer 116:4256–4265CrossRefPubMedGoogle Scholar
  11. Patel SB, Stenehjem DD, Gill DM, Tantravahi SK, Agarwal AM, Hsu J, Vuong W, Pal SK, Agarwal N (2012) Everolimus versus temsirolimus in metastatic renal cell carcinoma after progression with previous systemic therapies. Clin Genitourin Cancer 14:153–159CrossRefGoogle Scholar
  12. Ryan J, PA P (2013) Cyclodextrin-based polymers for therapeutics delivery. US Patent 2012-0065161. Filled 12 Aug 2011, Issued 15 Mar 2012Google Scholar
  13. Sherman S, Amzal B, Calvo E, Wang X, Park J, Liu Z, Lin C, Casciano R (2015) An indirect comparison of everolimus versus axitinib in US patients with advanced renal cell carcinoma in whom prior sunitinib therapy failed. Clin Ther 37:2552–2559CrossRefGoogle Scholar
  14. Shirotake S, Yasumizu Y, Ito K, Masunaga A, Ito Y, Miyazaki Y, Hagiwara M, Kanao K, Mikami S, Nakagawa K, Momma T, Masuda T, Asano T, Oyama M, Tanaka N, Mizuno R, Oya M (2016) Impact of second-line targeted therapy dose intensity on patients with metastatic renal cell carcinoma. Clin Genitourin Cancer 14:e575–e583CrossRefPubMedGoogle Scholar
  15. Stenner-Liewen F, Grünwald V, Greil R, Porta C (2013) The clinical potential of temsirolimus in second or later lines of treatment for metastatic renal cell carcinoma. Expert Rev Anticancer Ther 13:1021–1033CrossRefPubMedGoogle Scholar
  16. Tomalova B, Sirova M, Rossmann P, Pola R, Strohalm J, Chytil P, Cerny V, Tomala J, Kabesova M, Rihova B, Ulbrich K, Etrych T, Kovar M (2016) The structure-dependent toxicity, pharmacokinetics and anti-tumor activity of HPMA copolymer conjugates in the treatment of solid tumors and leukemia. J Control Release 223:1–10CrossRefPubMedGoogle Scholar
  17. Wang HB, Kuang H, Zhang W, Zhou X, Zhu T (2016) Synthesis and evaluation of a water-soluble epothilone B prodrug with remarkably decreased toxicity. Lett Org Chem 13:619–624CrossRefGoogle Scholar
  18. Wang H, Zheng X, Cai Z, Yu O, Zheng S, Zhu T (2017) Synthesis and evaluation of an injectable everolimus prodrug. Bioorg Med Chem Lett 27:1175–1178CrossRefPubMedGoogle Scholar
  19. Weikert S, Kempkensteffen C, Busch J, Johannsen M, Grünwald V, Zimmermann K, Flörcken A, Westermann J, Weinkauf L, Miller K, Keilholz U (2013) Sequential mTOR inhibitor treatment with temsirolimus in metastatic renal cell carcinoma following failure of VEGF receptor tyrosine kinase inhibitors. World J Urol 31:805–809CrossRefPubMedGoogle Scholar
  20. Yeo SH, Nihira T, Yamada Y (1998) Screening and identification of a novel lipase from Burkholderia sp. YY62 which hydrolyzes t-butyl esters effectively. J Gen Appl Microbiol 44:147–152CrossRefPubMedGoogle Scholar
  21. Zhao H, Rubio B, Sapra P, Wu D, Reddy P, Sai P, Martinez A, Gao Y, Lozanguiez Y, Longley C, Greenberger LM, Horak ID (2008) Novel prodrugs of SN38 using multiarm poly(ethylene glycol) linkers. Bioconjugate Chem 19:849–859CrossRefGoogle Scholar
  22. Zhu T, Shah SM, Saunders RW (2002) Water soluble SDZ-RAD esters. EP Patent 2002-1210350. Filled 16 Aug 2000, Issued 06 May 2002Google Scholar

Copyright information

© Springer Science+Business Media, LLC 2017

Authors and Affiliations

  1. 1.Zhejiang Hongyuan pharmaceutical Co., Ltd.LinhaiChina
  2. 2.Zhejiang Hisun pharmaceutical Co., Ltd.TaizhouChina

Personalised recommendations