Treatment of Allergic Asthma by Targeting Janus Kinase 3-Dependent Leukotriene Synthesis in Mast Cells with 4-(3′,5′-Dibromo-4′-hydroxyphenyl)amino-6,7-dimethoxyquinazoline (WHI-P97)
- Ravi Malaviya1,1,
- Chun-Lin Chen5,
- Christopher Navara6,
- Rama Malaviya,
- Xing-Ping Liu3,
- Margaret Keenan1,
- Barbara Waurzyniak4 and
- Fatih M. Uckun2,6
- Departments of 1Allergy and Inflammatory Diseases (R.M., R.M., M.K.), 2Immunology (F.M.U.), 3Chemistry (X.-P.L.), 4Experimental Pathology (B.W.), 5Pharmaceutical Sciences (C.-L.C.), and 6Drug Discovery Program (C.N., F.M.U.), Parker Hughes Institute, St. Paul, Minnesota
Abstract
4-(3′,5′-Dibromo-4′-hydroxyphenyl)amino-6,7-dimethoxyquinazoline (WHI-P97) is a rationally designed potent inhibitor of Janus kinase (JAK)-3. Treatment of mast cells with WHI-P97 inhibited the translocation of 5-lipoxygenase (5-LO) from the nucleoplasm to the nuclear membrane and consequently 5-LO-dependent leukotriene (LT) synthesis after IgE receptor/FcεRI crosslinking by >90% at low micromolar concentrations. WHI-P97 did not directly inhibit the enzymatic activity of 5-LO, but prevented its translocation to the nuclear membrane without affecting the requisite calcium signal. WHI-P97 was very well tolerated in mice, with no signs of toxicity at dose levels ranging from 5 μg/kg to 50 mg/kg, and LD10was not reached at a 50 mg/kg dose level when administered as a single i.p. or i.v. bolus dose. Therapeutic WHI-P97 concentrations, which inhibit mast cell leukotriene synthesis in vitro, could easily be achieved in vivo after the i.v. or i.p. administration of a single nontoxic 40 mg/kg bolus dose of WHI-P97. Notably, WHI-P97 showed promising biological activity in a mouse model of allergic asthma at nontoxic dose levels. Treatment of ovalbumin-sensitized mice with WHI-P97 prevented the development of airway hyper-responsiveness to methacholine in a dose-dependent fashion. Furthermore, WHI-P97 inhibited the eosinophil recruitment to the airway lumen after the ovalbumin challenge in a dose-dependent fashion. Further development of WHI-P97 may therefore provide the basis for new and effective treatment as well as prevention programs for allergic asthma in clinical settings.
Footnotes
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Send reprint requests to: Dr. Fatih M. Uckun, Parker Hughes Institute, 2665 Long Lake Rd., Suite 300, St. Paul, MN 55113. E-mail:fatih_uckun{at}mercury.ih.org
- Abbreviations:
- LT
- leukotriene
- WHI-P97
- 4-(3′,5′-dibromo-4′-hydroxyphenyl)amino-6,7-dimethoxyquinazoline
- WHI-P131
- 4-(4′-hydroxyphenyl)amino-6,7-dimethoxyquinazoline
- WHI-P112
- 4-(2′,5′-dibromophenyl)amino-6,7-dimethoxyquinazoline
- 5-LO
- 5-lipoxygenase
- FLAP
- 5-LO-activating protein, 5-HPETE, 5-hydroperoxyeicosatetraenoic acid
- 5-HETE
- 5-hydroxyeicosatetraenoic acid
- TX
- thromboxane
- PG
- prostaglandin
- PMSF
- phenylmethylsulfonyl fluoride
- JAK3
- Janus kinase 3
- DNP
- dinitrophenyl
- BMMC
- bone marrow mast cells
- TEA
- triethylamine
- ELISA
- enzyme-linked immunosorbent assay
- OVA
- ovalbumin
- Penh
- enhanced pause
- COX
- cyclooxygenase
- PIPES
- 1,4-piperazinediethanesulfonic acid
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- Received July 12, 2000.
- Accepted August 11, 2000.
- The American Society for Pharmacology and Experimental Therapeutics
