Efficient brain targeting and therapeutic intracranial activity of bortezomib through intranasal co-delivery with NEO100 in rodent glioblastoma models

Restricted access

OBJECTIVE

Many pharmaceutical agents are highly potent but are unable to exert therapeutic activity against disorders of the central nervous system (CNS), because the blood-brain barrier (BBB) impedes their brain entry. One such agent is bortezomib (BZM), a proteasome inhibitor that is approved for the treatment of multiple myeloma. Preclinical studies established that BZM can be effective against glioblastoma (GBM), but only when the drug is delivered via catheter directly into the brain lesion, not after intravenous systemic delivery. The authors therefore explored alternative options of BZM delivery to the brain that would avoid invasive procedures and minimize systemic exposure.

METHODS

Using mouse and rat GBM models, the authors applied intranasal drug delivery, where they co-administered BZM together with NEO100, a highly purified, GMP-manufactured version of perillyl alcohol that is used in clinical trials for intranasal therapy of GBM patients.

RESULTS

The authors found that intranasal delivery of BZM combined with NEO100 significantly prolonged survival of tumor-bearing animals over those that received vehicle alone and also over those that received BZM alone or NEO100 alone. Moreover, BZM concentrations in the brain were higher after intranasal co-delivery with NEO100 as compared to delivery in the absence of NEO100.

CONCLUSIONS

This study demonstrates that intranasal delivery with a NEO100-based formulation enables noninvasive, therapeutically effective brain delivery of a pharmaceutical agent that otherwise does not efficiently cross the BBB.

ABBREVIATIONS BBB = blood-brain barrier; BZM = bortezomib; cGMP = current good manufacturing practice; CNS = central nervous system; CSF = cerebrospinal fluid; GBM = glioblastoma; HPLC = high-performance liquid chromatography; IS = internal standard; MTT = methylthiazoletetrazolium; NEO100 = enriched perillyl alcohol manufactured under cGMP conditions; POH = perillyl alcohol; TMZ = temozolomide; USC = University of Southern California.

Article Information

Correspondence Thomas C. Chen: University of Southern California, Los Angeles, CA. tcchen@usc.edu.

INCLUDE WHEN CITING Published online March 15, 2019; DOI: 10.3171/2018.11.JNS181161.

Disclosures Dr. Chen reports an ownership interest (founder and stakeholder) in NeOnc Technologies, Los Angeles, CA.

© AANS, except where prohibited by US copyright law.

Headings

Figures

  • View in gallery

    Cytotoxic potency of BZM against glioma cell lines. Three different established glioma cell lines derived from mouse (GL26), rat (RG2), and human (U251) were seeded into 96-well plates and exposed to different concentrations of BZM. After 48 hours, standard MTT assay was performed to determine viability of cells. Percent survival was determined as compared to vehicle-treated cells (n ≥ 3, mean ± SD [error bars]).

  • View in gallery

    Concentrations of perillyl alcohol and perillic acid in rat brain. At different time points after intranasal delivery of NEO100 to Fisher rats, brains were collected for analysis. Two animals were used for each time point. A: One brain was used to determine POH concentrations in different brain regions (R1–R5; see schematic). B: The second brain was used to determine overall POH concentrations, as well as concentrations of its metabolite perillic acid.

  • View in gallery

    Concentrations of BZM in rat brain. A: Three rats received BZM via bolus tail vein injection. After 1 hour, serum and brains were collected. Tissue concentrations of BZM were determined as described in Methods. B: Rats with intracranially implanted RG2 glioma cells were administered with intranasal BZM in the presence or absence of NEO100. At different times thereafter, 3 animals per time point were euthanized. Their brains were collected and separated into tumor tissue and normal tissue (contralateral hemisphere). BZM concentrations were analyzed. Means and SDs (error bars) are shown.

  • View in gallery

    Effect of intranasal BZM plus NEO100 on survival of tumor-bearing mice. Human U251 GBM cells were implanted into the brains of athymic nude mice. Ten days after implantation, the animals were separated into different groups (5 animals per group) and treated with vehicle or drugs. Shown here are Kaplan-Meier survival curves. A: Animal survival in response to intravenous (IV) delivery of vehicle, BZM, or BZM plus NEO100. B: Animal survival in response to intranasal (IN) delivery of vehicle, BZM, NEO100, or BZM plus NEO100. In all cases, the dosage of BZM was 1 mg/kg per cycle, and the concentration of NEO100 was 0.3%. Animals were monitored for behavioral signs of neurological toxicity, which is also relevant in view of BZM’s known neurotoxic potential.23 Such signs emerged on occasion in a few animals but were aligned with late-stage intracranial tumor growth, not with BZM treatment. Figure is available in color online only.

  • View in gallery

    BZM content in CSF versus serum and stability. A: Six rats per time point were treated with intranasal BZM (3 with co-administered NEO100, 3 with vehicle). CSF and serum were collected, processed, and analyzed for BZM content. B: BZM was mixed with 0.3% NEO100, or vehicle only, and incubated at 37°C. Aliquots were removed and processed at different times, as indicated, to measure BZM content. Starting concentration of BZM was set at 100%. Shown in both figure parts are the means of the triplicate samples and the standard errors (error bars).

References

1

Agrawal MSaraf SSaraf SAntimisiaris SGChougule MBShoyele SA: Nose-to-brain drug delivery: An update on clinical challenges and progress towards approval of anti-Alzheimer drugs. J Control Release 281:1391772018

2

Aiello-Laws LRutledge DN: Management of adult patients receiving intraventricular chemotherapy for the treatment of leptomeningeal metastasis. Clin J Oncol Nurs 12:4294352008

3

Bota DAAlexandru DKeir STBigner DVredenburgh JFriedman HS: Proteasome inhibition with bortezomib induces cell death in GBM stem-like cells and temozolomide-resistant glioma cell lines, but stimulates GBM stem-like cells’ VEGF production and angiogenesis. J Neurosurg 119:141514232013

4

Chen TCDa Fonseca COSchönthal AH: Perillyl alcohol and its drug-conjugated derivatives as potential novel methods of treating brain metastases. Int J Mol Sci 17:172016

5

Chen TCFonseca COSchönthal AH: Preclinical development and clinical use of perillyl alcohol for chemoprevention and cancer therapy. Am J Cancer Res 5:158015932015

6

Chen TCWang WGolden EBThomas SSivakumar WHofman FM: Green tea epigallocatechin gallate enhances therapeutic efficacy of temozolomide in orthotopic mouse glioblastoma models. Cancer Lett 302:1001082011

7

Crowe TPGreenlee MHWKanthasamy AGHsu WH: Mechanism of intranasal drug delivery directly to the brain. Life Sci 195:44522018

8

da Fonseca COSchwartsmann GFischer JNagel JFuturo DQuirico-Santos T: Preliminary results from a phase I/II study of perillyl alcohol intranasal administration in adults with recurrent malignant gliomas. Surg Neurol 70:2592672008

9

da Fonseca COSimão MLins IRCaetano ROFuturo DQuirico-Santos T: Efficacy of monoterpene perillyl alcohol upon survival rate of patients with recurrent glioblastoma. J Cancer Res Clin Oncol 137:2872932011

10

da Fonseca COTeixeira RMSilva JCTde Saldanha da Gama Fischer JMeirelles OCLandeiro JA: Long-term outcome in patients with recurrent malignant glioma treated with perillyl alcohol inhalation. Anticancer Res 33:562556312013

11

Dhuria SVHanson LRFrey WH II: Intranasal delivery to the central nervous system: mechanisms and experimental considerations. J Pharm Sci 99:165416732010

12

Djupesland PGMessina JCMahmoud RA: The nasal approach to delivering treatment for brain diseases: an anatomic, physiologic, and delivery technology overview. Ther Deliv 5:7097332014

13

Dréan AGoldwirt LVerreault MCanney MSchmitt CGuehennec J: Blood-brain barrier, cytotoxic chemotherapies and glioblastoma. Expert Rev Neurother 16:128513002016

14

Fribley AZeng QWang CY: Proteasome inhibitor PS-341 induces apoptosis through induction of endoplasmic reticulum stress-reactive oxygen species in head and neck squamous cell carcinoma cells. Mol Cell Biol 24:969597042004

15

Friday BBAnderson SKBuckner JYu CGiannini CGeoffroy F: Phase II trial of vorinostat in combination with bortezomib in recurrent glioblastoma: a north central cancer treatment group study. Neuro Oncol 14:2152212012

16

Gabay MPThakkar JPStachnik JMWoelich SKVillano JL: Intra-CSF administration of chemotherapy medications. Cancer Chemother Pharmacol 70:1152012

17

Gross EASwenberg JAFields SPopp JA: Comparative morphometry of the nasal cavity in rats and mice. J Anat 135:83881982

18

Harkema JRCarey SAWagner JG: The nose revisited: a brief review of the comparative structure, function, and toxicologic pathology of the nasal epithelium. Toxicol Pathol 34:2522692006

19

Hemeryck AGeerts RMonbaliu JHassler SVerhaeghe TDiels L: Tissue distribution and depletion kinetics of bortezomib and bortezomib-related radioactivity in male rats after single and repeated intravenous injection of 14 C-bortezomib. Cancer Chemother Pharmacol 60:7777872007

20

Joshi SMeyers PMOrnstein E: Intracarotid delivery of drugs: the potential and the pitfalls. Anesthesiology 109:5435642008

21

Kardosh AGolden EBPyrko PUddin JHofman FMChen TC: Aggravated endoplasmic reticulum stress as a basis for enhanced glioblastoma cell killing by bortezomib in combination with celecoxib or its non-coxib analogue, 2,5-dimethyl-celecoxib. Cancer Res 68:8438512008

22

Labussiere MPinel SDelfortrie SPlenat FChastagner P: Proteasome inhibition by bortezomib does not translate into efficacy on two malignant glioma xenografts. Oncol Rep 20:128312872008

23

Meregalli C: An overview of bortezomib-induced neurotoxicity. Toxics 3:2943032015

24

Nawrocki STCarew JSPino MSHighshaw RADunner K JrHuang P: Bortezomib sensitizes pancreatic cancer cells to endoplasmic reticulum stress-mediated apoptosis. Cancer Res 65:11658116662005

25

Odia YKreisl TNAregawi DInnis EKFine HA: A phase II trial of tamoxifen and bortezomib in patients with recurrent malignant gliomas. J Neurooncol 125:1911952015

26

Ostermann SCsajka CBuclin TLeyvraz SLejeune FDecosterd LA: Plasma and cerebrospinal fluid population pharmacokinetics of temozolomide in malignant glioma patients. Clin Cancer Res 10:372837362004

27

Portnow JBadie BChen MLiu ABlanchard SSynold TW: The neuropharmacokinetics of temozolomide in patients with resectable brain tumors: potential implications for the current approach to chemoradiation. Clin Cancer Res 15:709270982009

28

Quintana DSWestlye LTRustan OGTesli NPoppy CLSmevik H: Low-dose oxytocin delivered intranasally with Breath Powered device affects social-cognitive behavior: a randomized four-way crossover trial with nasal cavity dimension assessment. Transl Psychiatry 5:e6022015

29

Raizer JJChandler JPFerrarese RGrimm SALevy RMMuro K: A phase II trial evaluating the effects and intra-tumoral penetration of bortezomib in patients with recurrent malignant gliomas. J Neurooncol 129:1391462016

30

Rambabu CVenkatrao SRamu GGanesh M: Estimation of bortezomib in bulk and its pharmaceutical dosage forms by using a novel validated accurate reverse phase high performance liquid chromatography. Int J Pharm Pharm Sci 3:3033052011

31

Reger MAWatson GSGreen PSWilkinson CWBaker LDCholerton B: Intranasal insulin improves cognition and modulates beta-amyloid in early AD. Neurology 70:4404482008

32

Richardson PGMitsiades CHideshima TAnderson KC: Proteasome inhibition in the treatment of cancer. Cell Cycle 4:2902962005

33

Scott KHayden PJWill AWheatley KCoyne I: Bortezomib for the treatment of multiple myeloma. Cochrane Database Syst Rev 4:CD0108162016

34

Serwer LPJames CD: Challenges in drug delivery to tumors of the central nervous system: an overview of pharmacological and surgical considerations. Adv Drug Deliv Rev 64:5905972012

35

Shingaki THidalgo IJFurubayashi TKatsumi HSakane TYamamoto A: The transnasal delivery of 5-fluorouracil to the rat brain is enhanced by acetazolamide (the inhibitor of the secretion of cerebrospinal fluid). Int J Pharm 377:85912009

36

Stupp RMason WPvan den Bent MJWeller MFisher BTaphoorn MJ: Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma. N Engl J Med 352:9879962005

37

Styczynski JOlszewska-Slonina DKolodziej BNapieraj MWysocki M: Activity of bortezomib in glioblastoma. Anticancer Res 26 (6B):449945032006

38

van Tellingen OYetkin-Arik Bde Gooijer MCWesseling PWurdinger Tde Vries HE: Overcoming the blood-brain tumor barrier for effective glioblastoma treatment. Drug Resist Updat 19:1122015

39

Wang DGao YYun L: Study on brain targeting of raltitrexed following intranasal administration in rats. Cancer Chemother Pharmacol 57:971042006

40

Wang FJiang XLu W: Profiles of methotrexate in blood and CSF following intranasal and intravenous administration to rats. Int J Pharm 263:172003

41

Wang WCho HYRosenstein-Sisson RMarín Ramos NIPrice RHurth K: Intratumoral delivery of bortezomib: impact on survival in an intracranial glioma tumor model. J Neurosurg 128:6957002018

42

Zhang YZhu XHou KZhao JHan ZZhang X: Mcl-1 downregulation sensitizes glioma to bortezomib-induced apoptosis. Oncol Rep 33:227722842015

TrendMD

Metrics

Metrics

All Time Past Year Past 30 Days
Abstract Views 68 68 68
Full Text Views 21 21 21
PDF Downloads 8 8 8
EPUB Downloads 0 0 0

PubMed

Google Scholar