Rho kinase inhibitor explained

Rho-kinase inhibitors (rho-associated protein kinase inhibitor or ROCK inhibitor) are a series of compounds that target rho kinase (ROCK) and inhibit the ROCK pathway.^[1] Clinical trials have found that inhibition of the ROCK pathway contributes to the cardiovascular benefits of statin therapy. Furthermore, ROCK inhibitors may have clinical applications for anti-erectile dysfunction, antihypertension, and tumor metastasis inhibition.^[2] More recently they have been studied for the treatment of glaucoma^[3] and as a therapeutic target for the treatment of cardiovascular diseases, including ischemic stroke.^[4] While statin therapy has been demonstrated to reduce the risk of major cardiovascular events, including ischemic stroke,^[5] the interplay between the ROCK pathway and statin therapy to treat and prevent strokes in older adults has not yet been proven.

On a cellular level, ROCK has multiple functions, including regulation of smooth muscle cell contraction, cell migration, and maintenance of cell viability and morphology, in part by regulating stress fibers and focal adhesions.^[2] Particularly, ROCK inhibitor is used for cell culture practice, in part to limit cellular death and limited dedifferentiation,^{[6] [7] [8]} and therefore widely adopted for induced pluripotent stem cells (iPSC) and embryonic stem cell cultures,^[9] although studies have shown mixed results for other cells types.^[10]

Molecular mechanism

Rho kinase inhibitors act on Rho kinase by altering the conformation of the protein, disrupting translocation to the plasma membrane, preventing ATP-dependent phosphorylation, and blocking RhoA binding to ROCK.^[11] Some studies suggest that Rho kinase inhibitors also play a role in anti-angiogenesis by blocking ERK and Akt signaling pathways.^{[12] [13]} Rho kinase inhibitor also functions by blocking rho-mediated dephosphorylation of MLC20.^[14]

Examples

A number of Rho kinase inhibitors are known.^{[15] [16] [17]}

• AT-13148^[18]
• BA-210
• β-Elemene
• Belumosudil
• Chroman 1^{[19] [20]}
• DJ4, which is a selective multi-specific ATP competitive inhibitor of activity of ROCK1 (IC50 of 5 nM), ROCK2 (IC50 of 50 nM), MRCKα (IC50 of 10 nM) and MRCKβ (IC50 of 100 nM) kinases without affecting activity of PAK1 and DMPK at 5 μM concentrations in in vitro cell-free kinase activity assays.^[21] DJ4 reduces stress fiber formation, and inhibits migration and invasion of melanoma (A375), breast (MD-MB-231), and non-small cell lung cancer (A549) cells independent of cell death induction.
• Fasudil, not currently approved by the U.S. Food and Drug Administration (FDA), but in Japan and China it is used for improvement and prevention of cerebral vasospasm and the cerebral ischemic symptoms subsequent to subarachnoid hemorrhage surgery.
• GSK-576371^[22]
• GSK429286A, C₂₁H₁₆F₄N₄O₂^[23]
• H-1152 Inhibitor H-1152 is approximately 9 to 16 times more potent than Y-27632 and HA-1077 (Fasudil)^[24]
• Hydroxyfasudil, an active metabolite of fasudil
  • Ibuprofen LX-7101^[25]
• Netarsudil, approved in United States for glaucoma and ocular hypertension.^[26]
• RKI-1447, a potent small molecule inhibitor of ROCK1 and ROCK2.^[27] Studies show that RKI-1447 could have anti-invasive and anti-tumor activities.
• Ripasudil, used in Japan for the treatment of glaucoma and ocular hypertension.
• TCS-7001
• Thiazovivin
• Verosudil (AR-12286) ^[28]
• Y-27632, the first small molecule ROCK inhibitor. It selectively inhibits ROCK1 with K_i of 140 nM.^[29]
• Y-30141
• Y-33075
• Y-39983

Further reading

• Rho Kinase (ROCK) Inhibitors and Their Therapeutic Potential. 2016

Notes and References

1. Liao. James K.. Seto. Minoru. Noma. Kensuke. July 2007. Rho Kinase (ROCK) Inhibitors. Journal of Cardiovascular Pharmacology. 50. 1. 17–24. 10.1097/FJC.0b013e318070d1bd. 0160-2446. 2692906. 17666911.
2. Liao JK, Seto M, Noma K . Rho kinase (ROCK) inhibitors . Journal of Cardiovascular Pharmacology . 50 . 1 . 17–24 . July 2007 . 17666911 . 2692906 . 10.1097/FJC.0b013e318070d1bd .
3. Wang SK, Chang RT . An emerging treatment option for glaucoma: Rho kinase inhibitors . Clinical Ophthalmology . 8 . 883–90 . 2014 . 24872673 . 4025933 . 10.2147/OPTH.S41000 . free .
4. Sladojevic N, Yu B, Liao JK . ROCK as a therapeutic target for ischemic stroke . Expert Review of Neurotherapeutics . 17 . 12 . 1167–1177 . December 2017 . 29057688 . 6221831 . 10.1080/14737175.2017.1395700 .
5. O'Brien EC, Greiner MA, Xian Y, Fonarow GC, Olson DM, Schwamm LH, Bhatt DL, Smith EE, Maisch L, Hannah D, Lindholm B, Peterson ED, Pencina MJ, Hernandez AF . 6 . Clinical Effectiveness of Statin Therapy After Ischemic Stroke: Primary Results From the Statin Therapeutic Area of the Patient-Centered Research Into Outcomes Stroke Patients Prefer and Effectiveness Research (PROSPER) Study . Circulation . 132 . 15 . 1404–13 . October 2015 . 26246175 . 10.1161/CIRCULATIONAHA.115.016183 . 11252336 . DaiWai Olson . free .
6. Matsumoto E, Furumatsu T, Kanazawa T, Tamura M, Ozaki T . ROCK inhibitor prevents the dedifferentiation of human articular chondrocytes . Biochemical and Biophysical Research Communications . 420 . 1 . 124–9 . March 2012 . 22405765 . 10.1016/j.bbrc.2012.02.127 .
7. Furumatsu T, Matsumoto-Ogawa E, Tanaka T, Lu Z, Ozaki T . ROCK inhibition enhances aggrecan deposition and suppresses matrix metalloproteinase-3 production in human articular chondrocytes . Connective Tissue Research . 55 . 2 . 89–95 . April 2014 . 24111521 . 10.3109/03008207.2013.852544 . 41211282 .
8. Qi Y, Liang X, Dai F, Guan H, Sun J, Yao W . RhoA/ROCK Pathway Activation is Regulated by AT1 Receptor and Participates in Smooth Muscle Migration and Dedifferentiation via Promoting Actin Cytoskeleton Polymerization . International Journal of Molecular Sciences . 21 . 15 . 5398 . July 2020 . 32751352 . 7432407 . 10.3390/ijms21155398 . free .
9. Claassen DA, Desler MM, Rizzino A . ROCK inhibition enhances the recovery and growth of cryopreserved human embryonic stem cells and human induced pluripotent stem cells . Molecular Reproduction and Development . 76 . 8 . 722–32 . August 2009 . 19235204 . 3257892 . 10.1002/mrd.21021 .
10. Nukaga T, Sakai D, Schol J, Suyama K, Nakai T, Hiyama A, Watanabe M . Minimal Sustainability of Dedifferentiation by ROCK Inhibitor on Rat Nucleus Pulposus Cells In Vitro . Spine Surgery and Related Research . 3 . 4 . 385–391 . 2019 . 31768460 . 6834460 . 10.22603/ssrr.2019-0019 .
11. 2006-03-01. Molecular Mechanism for the Regulation of Rho-Kinase by Dimerization and Its Inhibition by Fasudil. Structure. en. 14. 3. 589–600. 10.1016/j.str.2005.11.024. 0969-2126. Yamaguchi. Hiroto. Kasa. Miyuki. Amano. Mutsuki. Kaibuchi. Kozo. Hakoshima. Toshio. 16531242. free.
12. Hata. Yasuaki. Miura. Muneki. Nakao. Shintaro. Kawahara. Shuhei. Kita. Takeshi. Ishibashi. Tatsuro. 2008-02-01. Antiangiogenic properties of fasudil, a potent Rho-Kinase inhibitor. Japanese Journal of Ophthalmology. en. 52. 1. 16–23. 10.1007/s10384-007-0487-5. 18369695. 179423. 1613-2246.
13. Book: Khalil, Raouf A.. Rho Kinase in Vascular Smooth Muscle. 2010. Morgan & Claypool Life Sciences. en.
14. Nagumo. Hiromitsu. Sasaki. Yasuharu. Ono. Yoshitaka. Okamoto. Hiroyuki. Seto. Minoru. Takuwa. Yoh. 2000-01-01. Rho kinase inhibitor HA-1077 prevents Rho-mediated myosin phosphatase inhibition in smooth muscle cells. American Journal of Physiology. Cell Physiology. 278. 1. C57–C65. 10.1152/ajpcell.2000.278.1.C57. 10644512. 0363-6143. free.
15. You, Y., Zhu, K., Wang, J., Liang, Q., Li, W., Wang, L., ... & Shi, J. (2023). ROCK inhibitor: Focus on recent updates. Chinese Chemical Letters, 108336. https://doi.org/10.1016/j.cclet.2023.108336
16. Luo M, Li YQ, Lu YF, Wu Y, Liu R, Zheng YR, Yin M . Exploring the potential of RhoA inhibitors to improve exercise-recoverable spinal cord injury: A systematic review and meta-analysis . Journal of Chemical Neuroanatomy . 111 . 101879 . November 2020 . 33197553 . 10.1016/j.jchemneu.2020.101879 . 0891-0618 . free .
17. de Sousa GR, Vieira GM, das Chagas PF, Pezuk JA, Brassesco MS . Should we keep rocking? Portraits from targeting Rho kinases in cancer . Pharmacological Research . 160 . 105093 . October 2020 . 32726671 . 10.1016/j.phrs.2020.105093 . 220877441 .
18. Rath N, Munro J, Cutiongco MF, Jagiełło A, Gadegaard N, McGarry L, Unbekandt M, Michalopoulou E, Kamphorst JJ, Sumpton D, Mackay G, Vennin C, Pajic M, Timpson P, Olson MF . 6 . Rho Kinase Inhibition by AT13148 Blocks Pancreatic Ductal Adenocarcinoma Invasion and Tumor Growth . Cancer Research . 78 . 12 . 3321–3336 . June 2018 . 29669760 . 10.1158/0008-5472.CAN-17-1339 . 6005347 . free .
19. https://www.medchemexpress.com/chroman-1.html Chroman 1
20. Chen Y, Tristan CA, Chen L, Jovanovic VM, Malley C, Chu PH, Singeç, I . 6 . A versatile polypharmacology platform promotes cytoprotection and viability of human pluripotent and differentiated cells . Nature Methods . 18 . 5 . 528–541 . May 2021 . 33941937 . 10.1038/s41592-021-01126-2. 8314867 .
21. Kale VP, Hengst JA, Desai DH, Dick TE, Choe KN, Colledge AL, Takahashi Y, Sung SS, Amin SG, Yun JK . 6 . A novel selective multikinase inhibitor of ROCK and MRCK effectively blocks cancer cell migration and invasion . Cancer Letters . 354 . 2 . 299–310 . November 2014 . 25172415 . 4182185 . 10.1016/j.canlet.2014.08.032 .
22. Phrommintikul A, Tran L, Kompa A, Wang B, Adrahtas A, Cantwell D, Kelly DJ, Krum H . 6 . Effects of a Rho kinase inhibitor on pressure overload induced cardiac hypertrophy and associated diastolic dysfunction . American Journal of Physiology. Heart and Circulatory Physiology . 294 . 4 . H1804-14 . April 2008 . 18245565 . 10.1152/ajpheart.01078.2007 . 6554772 .
23. https://pubchem.ncbi.nlm.nih.gov/compound/gsk429286a GSK429286A@pubchem
24. 17603024 . 2007 . Teixeira . C. E. . Jin . L. . Priviero . F. B. . Ying . Z. . Webb . R. C. . Comparative pharmacological analysis of Rho-kinase inhibitors and identification of molecular components of Ca2+ sensitization in the rat lower urinary tract . Biochemical Pharmacology . 74 . 4 . 647–658 . 10.1016/j.bcp.2007.06.004 . 1987395 .
25. Boland S, Bourin A, Alen J, Geraets J, Schroeders P, Castermans K, Kindt N, Boumans N, Panitti L, Vanormelingen J, Fransen S, Van de Velde S, Defert O . 6 . Design, synthesis and biological characterization of selective LIMK inhibitors . Bioorganic & Medicinal Chemistry Letters . 25 . 18 . 4005–10 . September 2015 . 26233434 . 10.1016/j.bmcl.2015.07.009 .
26. Web site: RHOPRESSA presribing information . U.S. Food and Drug Administration .
27. Patel RA, Forinash KD, Pireddu R, Sun Y, Sun N, Martin MP, Schönbrunn E, Lawrence NJ, Sebti SM . 6 . RKI-1447 is a potent inhibitor of the Rho-associated ROCK kinases with anti-invasive and antitumor activities in breast cancer . Cancer Research . 72 . 19 . 5025–34 . October 2012 . 22846914 . 3463757 . 10.1158/0008-5472.CAN-12-0954 .
28. Williams RD, Novack GD, van Haarlem T, Kopczynski C . Ocular hypotensive effect of the Rho kinase inhibitor AR-12286 in patients with glaucoma and ocular hypertension . American Journal of Ophthalmology . 152 . 5 . 834–41.e1 . November 2011 . 21794845 . 10.1016/j.ajo.2011.04.012 .
29. Web site: information of different kinds of ROCK inhibitor. selleckchemicals.