ABSTRACT
The design and optimization of a novel trans-1,4-dioxycyclohexane GPR119 agonist series is described. A lead compound 21 was found to be a potent and efficacious GPR119 agonist across species, and possessed overall favorable pharmaceutical properties. Compound 21 demonstrated robust acute and chronic regulatory effects on glycemic parameters in the diabetic or non-diabetic rodent models.
Subject(s)
Cyclohexanes/chemistry , Cyclohexanes/therapeutic use , Diabetes Mellitus, Type 2/drug therapy , Hypoglycemic Agents/chemistry , Hypoglycemic Agents/therapeutic use , Receptors, G-Protein-Coupled/agonists , Administration, Oral , Animals , Blood Glucose/analysis , Cyclohexanes/administration & dosage , Cyclohexanes/pharmacokinetics , Diabetes Mellitus, Type 2/blood , Diabetes Mellitus, Type 2/metabolism , Humans , Hypoglycemic Agents/administration & dosage , Hypoglycemic Agents/pharmacokinetics , Male , Mice , Rats, Sprague-Dawley , Rats, Zucker , Receptors, G-Protein-Coupled/metabolism , Structure-Activity RelationshipABSTRACT
Modulators of S1P1 have proven utility for the treatment of autoimmune disease and efforts to identify new agents with improved safety and pharmacokinetic parameters are ongoing. Several new S1P1 chemotypes were designed and optimized for potency and oral bioavailability. These new agents are characterized by a 'tricyclic fused indole array' and are highly potent agonists of the S1P1 receptor.
Subject(s)
Drug Design , Indoles/chemistry , Receptors, Lysosphingolipid/agonists , Animals , Dogs , Half-Life , Humans , Indoles/chemical synthesis , Indoles/pharmacokinetics , Mice , Protein Binding , Protein Isoforms/agonists , Protein Isoforms/metabolism , Rats , Rats, Sprague-Dawley , Receptors, Lysosphingolipid/metabolism , Structure-Activity RelationshipABSTRACT
APD334 was discovered as part of our internal effort to identify potent, centrally available, functional antagonists of the S1P1 receptor for use as next generation therapeutics for treating multiple sclerosis (MS) and other autoimmune diseases. APD334 is a potent functional antagonist of S1P1 and has a favorable PK/PD profile, producing robust lymphocyte lowering at relatively low plasma concentrations in several preclinical species. This new agent was efficacious in a mouse experimental autoimmune encephalomyelitis (EAE) model of MS and a rat collagen induced arthritis (CIA) model and was found to have appreciable central exposure.
ABSTRACT
S1P1 is a validated target for treatment of autoimmune disease, and functional antagonists with superior safety and pharmacokinetic properties are being sought as second generation therapeutics. We describe the discovery and optimization of (7-benzyloxy-2,3-dihydro-1H-pyrrolo[1,2-a]indol-1-yl)acetic acids as potent, centrally available, direct acting S1P1 functional antagonists, with favorable pharmacokinetic and safety properties.
ABSTRACT
A series of 5-fluoro-4,6-dialkoxypyrimidine GPR119 modulators were discovered and optimized for in vitro agonist activity. A lead molecule was identified that has improved agonist efficacy relative to our clinical compound (APD597) and possesses reduced CYP2C9 inhibitory potential. This optimized lead was found to be efficacious in rodent models of glucose control both alone and in combination with a Dipeptidyl peptidase-4 (DPP-4) inhibitor.
Subject(s)
Drug Discovery , Piperidines/pharmacology , Pyridines/pharmacology , Receptors, G-Protein-Coupled/agonists , Dose-Response Relationship, Drug , Humans , Molecular Structure , Piperidines/chemical synthesis , Piperidines/chemistry , Pyridines/chemical synthesis , Pyridines/chemistry , Structure-Activity RelationshipABSTRACT
Two series of fused tricyclic indoles were identified as potent and selective S1P(1) agonists. In vivo these agonists produced a significant reduction in circulating lymphocytes which translated into robust efficacy in several rodent models of autoimmune disease. Importantly, these agonists were devoid of any activity at the S1P(3) receptor in vitro, and correspondingly did not produce S1P(3) mediated bradycardia in telemeterized rat.
Subject(s)
Immunologic Factors/chemistry , Indoles/chemistry , Receptors, Lysosphingolipid/agonists , Animals , Autoimmune Diseases/drug therapy , Disease Models, Animal , Female , Humans , Immunologic Factors/pharmacokinetics , Immunologic Factors/therapeutic use , Indoles/pharmacokinetics , Indoles/therapeutic use , Lymphocytes/immunology , Male , Mice , Mice, Inbred C57BL , Microsomes/metabolism , Rats , Rats, Sprague-Dawley , Receptors, Lysosphingolipid/metabolism , Structure-Activity RelationshipABSTRACT
S1P(1) receptor driven lymphopenia has proven utility in the treatment of an array of autoimmune disease states. As a part of our efforts to develop potent and selective S1P(1) receptor agonists, we have identified a novel chemical series of 4-oxo-4-(5-(5-phenyl-1,2,4-oxadiazol-3-yl)indolin-1-yl)butanoic acid S1P(1) receptor agonists.
Subject(s)
Autoimmune Diseases/drug therapy , Butyrates/chemical synthesis , Butyrates/pharmacology , Immunosuppressive Agents/chemical synthesis , Immunosuppressive Agents/pharmacology , Indoles/chemical synthesis , Indoles/pharmacology , Lymphocytes/metabolism , Nerve Tissue Proteins/agonists , Oxadiazoles/chemical synthesis , Oxadiazoles/pharmacology , RNA-Binding Proteins/agonists , Animals , Butyrates/pharmacokinetics , Disease Models, Animal , Dogs , Dose-Response Relationship, Drug , Drug Discovery , Drug Evaluation, Preclinical , Haplorhini , High-Throughput Screening Assays , Humans , Immunosuppressive Agents/pharmacokinetics , Indoles/pharmacokinetics , Male , Mice , Mice, Inbred BALB C , Nerve Tissue Proteins/chemistry , Oxadiazoles/pharmacokinetics , RNA-Binding Proteins/chemistry , Rats , Rats, Sprague-Dawley , Structure-Activity Relationship , Substrate SpecificityABSTRACT
A number of 2'- O-modified antisense oligonucleotides have been reported for their potential use in oligonucleotide-based therapeutics. To date, most of the in vivo data has been generated for 2'-O-MOE (2'-O-methoxyethyl)- and 2'-O-Me (2'-O-methyl)-modified ASOs (antisense oligonucleotides). We now report the synthesis and biological activity of another 2'-O-modification, namely 2'-O-[2-(methylamino)-2-oxoethyl] (2'-O-NMA). This modification resulted in an increase in the affinity of antisense oligonucleotides to complementary RNA similar to 2'-O-MOE-modified ASOs as compared to first-generation antisense oligodeoxynucleotides. The ASO modified with 2'-O-NMA reduced expression of PTEN mRNA in vitro and in vivo in a dose-dependent manner similar to 2'-O-MOE modified ASO. Importantly, toxicity parameters such as AST, ALT, organ weights, and body weights were found to be normal similar to 2'-O-MOE ASO-treated animal models. The data generated in these experiments suggest that 2'-O-NMA is a useful modification for potential application in both antisense and other oligonucleotide-based drug discovery efforts.
Subject(s)
Oligoribonucleotides, Antisense/chemical synthesis , Animals , Cell Line , Liver/cytology , Liver/metabolism , Male , Mice , Mice, Inbred BALB C , Nucleic Acid Hybridization , Oligoribonucleotides, Antisense/chemistry , Oligoribonucleotides, Antisense/pharmacology , PTEN Phosphohydrolase/biosynthesis , PTEN Phosphohydrolase/genetics , RNA, Messenger/biosynthesis , Structure-Activity RelationshipABSTRACT
[structure: see text] Oligonucleotides with novel modifications, 2'-O-[2-(amino)-2-oxoethyl] (2'-O-NAc), 2'-O-[2-(methylamino)-2-oxoethyl] (2'-O-NMAc), 2'-O-[2-(dimethylamino)-2-oxoethyl] (2'-O-DMAc), and 2'-O-[2-[[2-(dimethylamino)ethyl]amino]-2-oxoethyl] (2'-O-DMAEAc), have been synthesized. These modified oligonucleotides exhibit high binding affinity to complementary RNA (and not to DNA) and considerably enhance the nuclease stability of oligonucleotides with t(1/2) > 24 h.
Subject(s)
Oligonucleotides, Antisense/chemical synthesis , Drug Stability , Oligonucleotides, Antisense/chemistry , Oligonucleotides, Antisense/metabolism , Phosphodiesterase I , Phosphoric Diester Hydrolases/metabolism , RNA, Complementary/metabolism , Structure-Activity RelationshipABSTRACT
A novel 2'-modification, 2'-O-[2-(methylthio)ethyl] or 2'-O-MTE, has been incorporated into oligonucleotides and evaluated for properties relevant to antisense activity. The results were compared with the previously characterized 2'-O-[2-(methoxy)ethyl] 2'-O-MOE modification. As expected, the 2'-O-MTE modified oligonucleotides exhibited improved binding to human serum albumin compared to the 2'-O-MOE modified oligonucleotides. The 2'-O-MTE oligonucleotides maintained high binding affinity to target RNA. Nuclease digestion of 2'-O-MTE oligonucleotides showed that they have limited resistance to exonuclease degradation. We analyzed the crystal structure of a decamer DNA duplex containing the 2'-O-MTE modifcation. Analysis of the crystal structure provides insight into the improved RNA binding affinity, protein binding affinity and limited resistance of 2'-O-MTE modified oligonucleotides to exonuclease degradation.
Subject(s)
RNA/chemistry , Uridine/analogs & derivatives , Uridine/chemistry , Binding Sites , Crystallography, X-Ray , Nucleic Acid Heteroduplexes/chemistry , Nucleosides/chemistry , Oligodeoxyribonucleotides/chemistry , Oligodeoxyribonucleotides, Antisense/chemistry , Organophosphorus Compounds/chemistry , Protein BindingABSTRACT
A versatile synthetic route has been developed for the synthesis of 2'-O-[2-[(N,N-dimethylamino)oxy]ethyl] (abbreviated as 2'-O-DMAOE) modified purine and pyrimidine nucleosides and their corresponding nucleoside phosphoramidites and solid supports. To synthesize 2'-O-DMAOE purine nucleosides, the key intermediate B (Scheme 1) was obtained from the 2'-O-allyl purine nucleosides (13a and 15) via oxidative cleavage of the carbon-carbon bond to the corresponding aldehydes followed by reduction. To synthesize pyrimidine nucleosides, opening the 2,2'-anhydro-5-methyluridine 5 with the borate ester of ethylene glycol gave the key intermediate B. The 2'-O-(2-hydroxyethyl) nucleosides were converted, in excellent yield, by a regioselective Mitsunobu reaction, to the corresponding 2'-O-[2-[(1,3-dihydro-1,3-dioxo-2H-isoindol-2-yl)oxy]ethyl] nucleosides (18, 19, and 20). These compounds were subsequently deprotected and converted into the 2'-O-[2-[(methyleneamino)oxy]ethyl] derivatives (22, 23, and 24). Reduction and a second reductive amination with formaldehyde yielded the corresponding 2'-O-[2-[(N,N-dimethylamino)oxy]ethyl] nucleosides (25, 26, and 27). These nucleosides were converted to their 3'-O-phosphoramidites and controlled-pore glass solid supports in excellent overall yield. Using these monomers, modified oligonucleotides containing pyrimidine and purine bases were synthesized with phosphodiester, phosphorothioate, and both linkages (phosphorothioate and phosphodiester) present in the same oligonucleotide as a chimera in high yields. The oligonucleotides were characterized by HPLC, capillary gel electrophoresis, and ESMS. The effect of this modification on the affinity of the oligonucleotides for complementary RNA and on nuclease stability was evaluated. The 2'-O-DMAOE modification enhanced the binding affinity of the oligonucleotides for the complementary RNA (and not for DNA). The modified oligonucleotides that possessed the phosphodiester backbone demonstrated excellent resistance to nuclease with t(1/2) > 24 h.
