A year-long extended release nanoformulated cabotegravir prodrug

Abstract

Long-acting cabotegravir (CAB) extends antiretroviral drug administration from daily to monthly. However, dosing volumes, injection site reactions and health-care oversight are obstacles towards a broad usage. The creation of poloxamer-coated hydrophobic and lipophilic CAB prodrugs with controlled hydrolysis and tissue penetrance can overcome these obstacles. To such ends, fatty acid ester CAB nanocrystal prodrugs with 14, 18 and 22 added carbon chains were encased in biocompatible surfactants named NMCAB, NM2CAB and NM3CAB and tested for drug release, activation, cytotoxicity, antiretroviral activities, pharmacokinetics and biodistribution. Pharmacokinetics studies, performed in mice and rhesus macaques, with the lead 18-carbon ester chain NM2CAB, showed plasma CAB levels above the protein-adjusted 90% inhibitory concentration for up to a year. NM2CAB, compared with NMCAB and NM3CAB, demonstrated a prolonged drug release, plasma circulation time and tissue drug concentrations after a single 45 mg per kg body weight intramuscular injection. These prodrug modifications could substantially improve CAB’s effectiveness.

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Fig. 1: Synthesis and characterization of the CAB prodrugs.
Fig. 2: Synthesis and characterization of nanoformulations of CAB and prodrugs.
Fig. 3: In vitro characterization.
Fig. 4: CAB levels in plasma and tissues.
Fig. 5: Prodrug measurements.
Fig. 6: PK, BD and toxicological assessments in RMs.

Data availability

The data supporting the study’s findings are available within the article and its supplementary files or from the corresponding authors upon request. All the relevant data used to generate Figs. 4a,c,k, 5a–i and 6a–c are included as Source Data.

References

  1. 1.

    Fauci, A. S., Redfield, R. R., Sigounas, G., Weahkee, M. D. & Giroir, B. P. Ending the HIV epidemic: a plan for the United States. J. Am. Med. Assoc. 321, 844–845 (2019).

    Google Scholar 

  2. 2.

    Gendelman, H. E., McMillan, J., Bade, A. N., Edagwa, B. & Kevadiya, B. D. The promise of long-acting antiretroviral therapies: from need to manufacture. Trends Microbiol. 27, 593–606 (2019).

    CAS  Google Scholar 

  3. 3.

    Currier, J. S. Monthly injectable antiretroviral therapy—version 1.0 of a new treatment approach. N. Engl. J. Med. 382, 1164–1165 (2020).

    Google Scholar 

  4. 4.

    Margolis, D. A. et al. Long-acting intramuscular cabotegravir and rilpivirine in adults with HIV-1 infection (LATTE-2): 96-week results of a randomised, open-label, phase 2b, non-inferiority trial. Lancet 390, 1499–1510 (2017).

    CAS  Google Scholar 

  5. 5.

    Orkin, C. et al. Long-acting cabotegravir and rilpivirine after oral induction for HIV-1 Infection. N. Engl. J. Med. 382, 1124–1135 (2010).

    Google Scholar 

  6. 6.

    Swindells, S. et al. Long-acting cabotegravir and rilpivirine for maintenance of HIV-1 suppression. N. Engl. J. Med. 382, 1112–1123 (2020).

    CAS  Google Scholar 

  7. 7.

    ViiV Healthcare submits new drug application to US FDA for the first monthly, injectable, two-drug regimen of cabotegravir and rilpivirine for treatment of HIV (ViiV Healthcare, 2019); https://viivhealthcare.com/en-gb/media/press-releases/2019/april/viiv-healthcare-submits-new-drug-application-to-us-fda-for-the-first-monthly-injectable-two-drug-regimen-of-cabotegravir-and-rilpivirine-for-treatment-of-hiv/

  8. 8.

    ViiV Healthcare receives complete response letter from US FDA for use of investigational cabotegravir and rilpivirine long-acting regimen in the treatment of HIV (ViiV Healthcare, 2019); https://viivhealthcare.com/en-gb/media/press-releases/2019/december/complete-response-letter-from-us-fda/

  9. 9.

    Kovarova, M. et al. Ultra-long-acting removable drug delivery system for HIV treatment and prevention. Nat. Commun. 9, 4156 (2018).

    Google Scholar 

  10. 10.

    Gunawardana, M. et al. Pharmacokinetics of long-acting tenofovir alafenamide (GS-7340) subdermal implant for HIV prophylaxis. Antimicrob. Agents Chemother. 59, 3913–3919 (2015).

    CAS  Google Scholar 

  11. 11.

    Flexner, C. Antiretroviral implants for treatment and prevention of HIV infection. Curr. Opin. HIV AIDS 13, 374–380 (2018).

    CAS  Google Scholar 

  12. 12.

    Barrett, S. E. et al. Extended-duration MK-8591-eluting implant as a candidate for HIV treatment and prevention. Antimicrob. Agents Chemother. 62, e01058-18 (2018).

    Google Scholar 

  13. 13.

    Markowitz, M. et al. Safety and tolerability of long-acting cabotegravir injections in HIV-uninfected men (ECLAIR): a multicentre, double-blind, randomised, placebo-controlled, phase 2a trial. Lancet HIV 4, e331–e340 (2017).

    Google Scholar 

  14. 14.

    Zhou, T. et al. Creation of a nanoformulated cabotegravir prodrug with improved antiretroviral profiles. Biomaterials 151, 53–65 (2018).

    CAS  Google Scholar 

  15. 15.

    Dash, P. K. et al. Sequential LASER ART and CRISPR treatments eliminate HIV-1 in a subset of infected humanized mice. Nat. Commun. 10, 2753 (2019).

    Google Scholar 

  16. 16.

    Hilaire, J. R. et al. Creation of a long-acting rilpivirine prodrug nanoformulation. J. Control. Release 311–312, 201–211 (2019).

    Google Scholar 

  17. 17.

    Sillman, B. et al. Creation of a long-acting nanoformulated dolutegravir. Nat. Commun. 9, 443 (2018).

    Google Scholar 

  18. 18.

    Smith, N. et al. A long acting nanoformulated lamivudine ProTide. Biomaterials 223, 119476 (2019).

    CAS  Google Scholar 

  19. 19.

    Soni, D. et al. Synthesis of a long acting nanoformulated emtricitabine ProTide. Biomaterials 222, 119441 (2019).

    CAS  Google Scholar 

  20. 20.

    Huttunen, K. M., Raunio, H. & Rautio, J. Prodrugs—from serendipity to rational design. Pharm. Rev. 63, 750–771 (2011).

    CAS  Google Scholar 

  21. 21.

    Rautio, J. et al. Prodrugs: design and clinical applications. Nat. Rev. Drug Discov. 7, 255–270 (2008).

    CAS  Google Scholar 

  22. 22.

    Bahar, F. G., Ohura, K., Ogihara, T. & Imai, T. Species difference of esterase expression and hydrolase activity in plasma. J. Pharm. Sci. 101, 3979–3988 (2012).

    CAS  Google Scholar 

  23. 23.

    Malamatari, M., Taylor, K. M. G., Malamataris, S., Douroumis, D. & Kachrimanis, K. Pharmaceutical nanocrystals: production by wet milling and applications. Drug Discov. Today 23, 534–547 (2018).

    CAS  Google Scholar 

  24. 24.

    Zhou, T. et al. Optimizing the preparation and stability of decorated antiretroviral drug nanocrystals. Nanomedicine (Lond.) 13, 871–885 (2018).

    CAS  Google Scholar 

  25. 25.

    Darville, N. et al. Intramuscular administration of paliperidone palmitate extended-release injectable microsuspension induces a subclinical inflammatory reaction modulating the pharmacokinetics in rats. J. Pharm. Sci. 103, 2072–2087 (2014).

    CAS  Google Scholar 

  26. 26.

    Kadiu, I., Nowacek, A., McMillan, J. & Gendelman, H. E. Macrophage endocytic trafficking of antiretroviral nanoparticles. Nanomedicine (Lond.) 6, 975–994 (2011).

    CAS  Google Scholar 

  27. 27.

    Nowacek, A., Kadiu, I., McMillan, J. & Gendelman, H. E. Immunoisolation of nanoparticles containing endocytic vesicles for drug quantitation. Methods Mol. Biol. 991, 41–46 (2013).

    CAS  Google Scholar 

  28. 28.

    McMillan, J. et al. Pharmacokinetics of a long-acting nanoformulated dolutegravir prodrug in rhesus macaques. Antimicrob. Agents Chemother. 62, e01316-17 (2017).

    Google Scholar 

  29. 29.

    Rohani, S., Horne, S. & Murthy, K. Control of product quality in batch crystallization of pharmaceuticals and fine chemicals. Part 1: design of the crystallization process and the effect of solvent. Org. Process Res. Dev. 9, 858–872 (2005).

    CAS  Google Scholar 

  30. 30.

    Nair, A. B. & Jacob, S. A simple practice guide for dose conversion between animals and human. J. Basic Clin. Pharm. 7, 27–31 (2016).

    Google Scholar 

  31. 31.

    Kerrigan, D. et al. Experiences with long acting injectable ART: a qualitative study among PLHIV participating in a Phase II study of cabotegravir + rilpivirine (LATTE-2) in the United States and Spain. PLoS ONE 13, e0190487 (2018).

    Google Scholar 

  32. 32.

    Osterberg, L. & Blaschke, T. Adherence to medication. N. Engl. J. Med. 353, 487–497 (2005).

    CAS  Google Scholar 

  33. 33.

    Shubber, Z. et al. Patient-reported barriers to adherence to antiretroviral therapy: a systematic review and meta-analysis. PLoS Med. 13, e1002183 (2016).

    Google Scholar 

  34. 34.

    Edagwa, B., McMillan, J., Sillman, B. & Gendelman, H. E. Long-acting slow effective release antiretroviral therapy. Expert Opin. Drug Deliv. 14, 1281–1291 (2017).

    CAS  Google Scholar 

  35. 35.

    Trezza, C., Ford, S. L., Spreen, W., Pan, R. & Piscitelli, S. Formulation and pharmacology of long-acting cabotegravir. Curr. Opin. HIV AIDS 10, 239–245 (2015).

    CAS  Google Scholar 

  36. 36.

    Wang, D. et al. Human carboxylesterases: a comprehensive review. Acta Pharma. Sin. B 8, 699–712 (2018).

    Google Scholar 

  37. 37.

    McPherson, T. D., Sobieszczyk, M. E. & Markowitz, M. Cabotegravir in the treatment and prevention of human immunodeficiency virus-1. Expert Opin. Investig. Drugs 27, 413–420 (2018).

    CAS  Google Scholar 

  38. 38.

    Stellbrink, H. J. & Hoffmann, C. Cabotegravir: its potential for antiretroviral therapy and preexposure prophylaxis. Curr. Opin. HIV AIDS 13, 334–340 (2018).

    CAS  Google Scholar 

  39. 39.

    Rautio, J., Meanwell, N. A., Di, L. & Hageman, M. J. The expanding role of prodrugs in contemporary drug design and development. Nat. Rev. Drug Discov. 17, 559–587 (2018).

    CAS  Google Scholar 

  40. 40.

    Landovitz, R. J. et al. Safety, tolerability, and pharmacokinetics of long-acting injectable cabotegravir in low-risk HIV-uninfected individuals: HPTN 077, a phase 2a randomized controlled trial. PLoS Med. 15, e1002690 (2018).

    Google Scholar 

  41. 41.

    Murray, M. I. et al. Satisfaction and acceptability of cabotegravir long-acting injectable suspension for prevention of HIV: patient perspectives from the ECLAIR trial. HIV Clin. Trials 19, 129–138 (2018).

    CAS  Google Scholar 

  42. 42.

    Penrose, K. J. et al. Selection of rilpivirine-resistant HIV-1 in a seroconverter from the SSAT 040 trial who received the 300-mg dose of long-acting rilpivirine (TMC278LA). J. Infect. Dis. 213, 1013–1017 (2016).

    CAS  Google Scholar 

  43. 43.

    Bollinger R. C. et al. Addressing the global burden of hepatitis B virus while developing long-acting injectables for the prevention and treatment of HIV. Lancet HIV https://doi.org/10.1016/S2352-3018(19)30342-X (2019).

  44. 44.

    Ford, S. L. et al. Effect of rifampin on the single-dose pharmacokinetics of oral cabotegravir in healthy subjects. Antimicrob. Agents Chemother. 61, e00487-17 (2017).

    Google Scholar 

  45. 45.

    Rajoli, R. K. R. et al. Predicting drug–drug interactions between rifampicin and long-acting cabotegravir and rilpivirine using physiologically based pharmacokinetic modeling. J. Infect. Dis. 219, 1735–1742 (2019).

    Google Scholar 

  46. 46.

    Experimental HIV Vaccine Regimen Ineffective In preventing HIV (National Institute of Allergy and Infectious Diseases, 2020); https://www.nih.gov/news-events/news-releases/experimental-hiv-vaccine-regimen-ineffective-preventing-hiv.

  47. 47.

    Benitez-Gutierrez, L. et al. Treatment and prevention of HIV infection with long-acting antiretrovirals. Expert Rev. Clin. Pharm. 11, 507–517 (2018).

    CAS  Google Scholar 

  48. 48.

    de Mendoza, C. & Soriano, V. Tough requirements for new antiretroviral drugs. Lancet HIV 7, e150–e151 (2020).

    Google Scholar 

  49. 49.

    Smith, R. A. et al. In vitro antiviral activity of cabotegravir against HIV-2. Antimicrob. Agents Chemother. 62, e01299-18 (2018).

    Google Scholar 

  50. 50.

    Committee for the Update of the Guide for the Care and Use of Laboratory Animals Guide for the Care and Use of Laboratory Animals 8th edn (National Academies, 2011).

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Acknowledgements

We thank the University of Nebraska Medical Center (UNMC) cores for NMR (E. Ezell), elutriation and cell separation (M. Che, N. Ly and L. Wu), electron microscopy (N. Conoan and T. Bargar) and comparative medicine for technical assistance and animal care. We also thank S. Valloppilly of the University of Nebraska-Lincoln Nebraska Center for Materials and Nanoscience for X-ray structural characterization. We thank Z. You of the University of Nebraska-Lincoln Center for Biotechnology for SEM analysis of the nanoparticles. We thank A. Dash and D. Munt of Creighton University for the assistance in FTIR characterization and analysis. A special thank you to S. Cohen, UNMC, for the tissue histopathology assessments, and to P. K. Dash and J. Herskovitz, UNMC, for assistance in the execution and interpretation of experiments used in these and related works. R. Taylor, UNMC, is thanked for outstanding editorial support. This research is supported by the University of Nebraska Foundation, which includes donations from the Carol Swarts, M.D. Emerging Neuroscience Research Laboratory, the Margaret R. Larson Professorship, the Frances and Louie Blumkin Endowment and the Harriet Singer Endowment, the Vice Chancellor’s Office of the University of Nebraska Medical Center for Core Facilities and the National Institutes of Health grants 1R01AI145542–01A1, P01 DA028555, R01 NS36126, P01 NS31492, 2R01 NS034239, P01 MH64570, P01 NS43985, P30 MH062261, R01 AG043540 and 1 R56 AI138613–01A1. A special thank you is extended to J. Gold for his continuous support of translational research activities for our medical centre.

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Authors

Contributions

T.A.K. synthesized the prodrugs, performed the formulation characterization and cell-based laboratory experiments and interpreted the data, prepared the figure, and co-wrote the manuscript. A.N.B. designed and carried out the laboratory and rodent experiments, analysed and interpreted the data, supervised the project, prepared the figures and tables, and co-wrote and edited the manuscript. B.S., B.L.D.S., M.S.W., N.G., J.R.H. and S.S. performed the laboratory, physicochemical and/or rodent experiments. A.S. prepared NM2CAB in the Nebraska Nanomedicine Production Plant using good laboratory practice protocols. B.G.L., B.M.M. and H.S.F. performed and analysed the data from the non-human primate experiments. P.L.D., T.-Y.Y. and G.M. designed and executed the prodrug recrystallization, scale up and formulation manufacture experiments. Y.A. performed the data acquisition and interpretation. J.M.M. designed, supervised and analysed the mass spectrometry data. R.L.M. and J.M. performed statistical evaluations and data analyses. B.J.E. was responsible for the project conception, study and prodrug design, chemical drug synthesis and nanoformulation schemes, provided supervision, data analyses and interpretation, manuscript editing and funding acquisitions. H.E.G. was responsible for the project conception and study integration, integrated each of the study arms, supervised experimental design and data interpretation, co-wrote and edited the manuscript and provided funding acquisitions. All the authors critically evaluated the manuscript prior to submission.

Corresponding authors

Correspondence to Benson J. Edagwa or Howard E. Gendelman.

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Competing interests

B.J.E. and H.E.G. are named inventors on patents that cover the medicinal and polymer chemistry technologies employed in this article that encompass the synthesis of long-acting cabotegravir prodrugs and formulation manufacturing. H.E.G. is the Interim Director of the Nebraska Nanomedicine Production Plant, a good manufacturing programme facility. The authors declare that this work was produced solely by the authors and that no other individuals or entities influenced any aspects of the work including, but not limited to, the study conception and design, data acquisition, analyses and interpretation, and writing of the manuscript. No other entities provided funds for the work. The authors further declare that they have received no financial compensation from any other third parties for any aspects of the published work. The remaining authors declare no competing interests.

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Supplementary information

Supplemental Information

Supplementary Figs. 1–14, Tables 1–5 and Methods.

Reporting summary

Source data

Source Data Fig. 4

Raw CAB levels: Source Data Fig. 4a; Source Data Fig. 4c–k.

Source Data Fig. 5

Raw CAB prodrug levels: Source Data Fig. 5a–h; Source Data Fig. 5i.

Source Data Fig. 6

Raw CAB and prodrug levels in rhesus macaques: Source Data Fig. 6a; Source Data Fig. 6b,c.

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Kulkarni, T.A., Bade, A.N., Sillman, B. et al. A year-long extended release nanoformulated cabotegravir prodrug. Nat. Mater. (2020). https://doi.org/10.1038/s41563-020-0674-z

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