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. 2023 Jul 31;14(8):1054-1062.
doi: 10.1021/acsmedchemlett.3c00136. eCollection 2023 Aug 10.

Discovery of the TLR7/8 Antagonist MHV370 for Treatment of Systemic Autoimmune Diseases

Phil Alper  1 , Claudia Betschart  2 , Cédric André  2 , Thomas Boulay  2 , Dai Cheng  1 , Jonathan Deane  1 , Michael Faller  2 , Roland Feifel  2 , Ralf Glatthar  2 , Dong Han  1 , Rene Hemmig  2 , Tao Jiang  1 , Thomas Knoepfel  2 , Jillian Maginnis  1 , Daniel Mutnick  1 , Wei Pei  1 , Giulia Ruzzante  2 , Peter Syka  1 , Guobao Zhang  1 , Yi Zhang  1 , Florence Zink  2 , Géraldine Zipfel  2 , Stuart Hawtin  2 , Tobias Junt  2 , Pierre-Yves Michellys  1
Affiliations

Discovery of the TLR7/8 Antagonist MHV370 for Treatment of Systemic Autoimmune Diseases

Phil Alper et al. ACS Med Chem Lett. .

Abstract

Toll-like receptor (TLR) 7 and TLR8 are endosomal sensors of the innate immune system that are activated by GU-rich single stranded RNA (ssRNA). Multiple genetic and functional lines of evidence link chronic activation of TLR7/8 to the pathogenesis of systemic autoimmune diseases (sAID) such as Sjögren's syndrome (SjS) and systemic lupus erythematosus (SLE). This makes targeting TLR7/8-induced inflammation with small-molecule inhibitors an attractive approach for the treatment of patients suffering from systemic autoimmune diseases. Here, we describe how structure-based optimization of compound 2 resulted in the discovery of 34 (MHV370, (S)-N-(4-((5-(1,6-dimethyl-1H-pyrazolo[3,4-b]pyridin-4-yl)-3-methyl-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-1-yl)methyl)bicyclo[2.2.2]octan-1-yl)morpholine-3-carboxamide). Its in vivo activity allows for further profiling toward clinical trials in patients with autoimmune disorders, and a Phase 2 proof of concept study of MHV370 has been initiated, testing its safety and efficacy in patients with Sjögren's syndrome and mixed connective tissue disease.

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Conflict of interest statement

The authors declare no competing financial interest.

Figures

Figure 1
Figure 1
Role of TLR7 and TLR8 in the pathology of SLE and related chronic inflammatory diseases. ssRNA complexes are immune complexes of RNPs and autoantibodies or complexes of ssRNA with amphiphilic peptides.
Figure 2
Figure 2
Discovery of compound 2 by SAR database scouting TLR7 cellular assays in human PBMCs and mouse splenocytes; details in the Supporting Information. aIC50 ± SD, numbers in parentheses denote the number of independent experiments.
Scheme 1
Scheme 1. Synthesis of Compound 2
Reagents and conditions. a. Cs2CO3, DMSO, 120°C, 18 h. b. H-cube. c. Pd2dba3, RuPhos, Cs2CO3, THF, 80°C, 18 h. d. 4 N HCl/dioxane, MeOH, RT, 1h. e. Crystallization in IPA. f. Ambersep900OH, MeOH, rt, 1 h.
Scheme 2
Scheme 2. Synthetic Route of Alkyl-Substituted Amines
Reagents and conditions: a. Pd2dba3, Cs2CO3, RuPhos, dioxane, 10, reflux 18 h. b. HCl-dioxane, RT. c. Paraformaldehyde, DIEPA, NaBH3CN, THF/MeOH, RT. d. Cyclobutanone, NaBH3CN, AcOH, MeOH, RT to 75°C. e. Oxetan-3-one, NaBH3(OAc)3, DCE, AcOH, RT. f, AcCl or MsCl, DIPEA, DCM, 0°C to RT. g. 1-Bromo-2-methoxyethane, K2CO3, IPA, reflux. h/k. Corresponding dialkyl bromide, K2CO3, EtOH, 120°C, MW irradiation, 30 min. l. (S)-1-(4-Methylbenzenesulfonate)-2-(((R)-1-(4-methylbenzenesulfonate)-2-yl)oxy)propane, DIEPA, DMA, 100°C, 18h. m. 2,2-Difluoroethyl trifluoromethanesulfonate, DIPEA, THF, reflux, ON.
Scheme 3
Scheme 3. Synthetic Route for Compounds 23–30
Reagents and conditions: a. Corresponding amino acid (boc-protected where required), HATU, DIEPA, DCM, RT, then HCl-dioxane. b. (30) 2-bromo-N,N-dimethylacetamide, Cs2CO3, DMF, RT.
Scheme 4
Scheme 4. Synthetic Route for Compounds 31–35
Reagents and conditions: a. tert-Butyl methyl(2-oxoethyl)carbamate, NaBH(OAc)3, CPME, RT. b. 2-Bromoacetate, K2CO3, EtOH, MW irradiation, 150°C, 1 h then 1 N NaOH, RT. c. HCl-dioxane, RT. d. HATU, DIEPA, DMF, 0 °C. e 2-Bromoacetylbromide, DIEPA, DCM, 0°C to RT followed by HCl-dioxane, RT. f. K2CO3, EtOH, MW irradiation, 120 °C, 40 min followed 1 N aqueous HCl and lyophilization. g. Corresponding morpholino carboxylic acid, HATU, DIEPA, DCM, RT, then HCl-dioxane.
Figure 3
Figure 3
Cocrystal structure of compound 34 (MHV370) with TLR8 (PDB 8PFI). (a) Full view of the homodimeric structure and the two symmetrical binding sites formed between thae two TLR8 chains (one TLR8 monomer in light green and the second monomer in light blue), carbon atoms of compound 34 (MHV370) in orange. (b) Close-up view of one binding site showing amino acids in close contact with compound 34 (MHV370). Notable interactions include a hydrogen bond interaction (represented magenta dotted line) between the pyrazolo-pyridine moiety and Gly351 and many hydrophobic interactions between the central part of compound 34 (MHV370) and residues Phe261, Phe346, Val378, Ile403, Phe405, and Phe495.
Figure 4
Figure 4
In vivo efficacy of compound 34 (MHV370). Compound 34 (MHV370) suppresses serum levels of multiple proinflammatory cytokines in mice after an acute TLR7 challenge. Bars represent means and SD, dots represent individual mice. Blue numbers represent blood exposures in nM (±SD), ***p > 0.001, ****p > 0.0001, ANOVA with Dunnett’s post test.
See this image and copyright information in PMC

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