OpenSourceMalaria:Triazolopyrazine (TP) Series

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=Open Source Malaria Series 4: The Triazolopyrazine (TP) Series=
=Open Source Malaria Series 4: The Triazolopyrazine (TP) Series=
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==What's New==
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 +
This series is currently '''active'''. (How to respond/input, for example if you want to suggest a molecule that should be made, is described in the [http://opensourcemalaria.org/ Landing Page] under "Join the Team")
 +
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[https://github.com/OpenSourceMalaria/OSM_To_Do_List/issues?labels=Series+4&milestone=&page=1&state=open Automatically updated list of to do items in this Series]
 +
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[https://github.com/OpenSourceMalaria/OSM_To_Do_List/issues?labels=Being+Synthesised+Now%2CSeries+4&page=1&state=open Automatically updated list of compounds being made in this series]
 +
 +
[http://malaria.ourexperiment.org/osddmalaria_meeting_/9470 Most recent online meeting relevant to this series]
 +
 +
Most recent PDF newsletter may be downloaded using [http://malaria.ourexperiment.org/data/files/10840/OSM%20Newsletter%20April%202014.pdf this link].
==Introduction==
==Introduction==
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===Preamble===
===Preamble===
-
The Triazolopyrazine series is the newest of the OSM series. It was announced on September 10th 2013 as Series 4.
+
The Triazolopyrazine Series is the newest of the OSM series. It was [http://www.mmv.org/newsroom/news/potential-new-class-antimalarials-now-open-source announced] on September 10th 2013 and is sometimes referred to as the ''TP Series'', or ''OSM Series 4''.
 +
 
 +
[[Image:Representative TP Compound.png|thumb|center|250px|Representative Series 4 Compound]]
 +
 
 +
The series arises from industrial work that cannot be fully disclosed which was followed by some hit-to-lead work funded directly by MMV and performed by a CRO which can.
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The series arises from industrial work that cannot be fully disclosed which was followed by some hit-to-lead work funded directly by MMV which can.
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A great deal of exploration of the series has been done, with significant diversity in the core and pendant groups. The series includes many potent compounds.
There is evidence from parasite ion regulation assays (below) that these compounds may be PfATP4 inhibitors. Such evidence distinguishes Series 4 from Series 1-3 where there was no experimental evidence for a mechanism of action.
There is evidence from parasite ion regulation assays (below) that these compounds may be PfATP4 inhibitors. Such evidence distinguishes Series 4 from Series 1-3 where there was no experimental evidence for a mechanism of action.
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As with everything involved in OSM, suggestions can be given in [http://opensourcemalaria.org/# multiple ways].
As with everything involved in OSM, suggestions can be given in [http://opensourcemalaria.org/# multiple ways].
-
===What is Known at the Start of the Campaign===
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===Start of the Campaign===
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A [http://malaria.ourexperiment.org/uri/404# briefing document] written for MMV constitutes what was known at the start of the open project.
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A [http://malaria.ourexperiment.org/uri/404# briefing document] written for MMV was the initial knowledgebase. This was accompanied by a [http://malaria.ourexperiment.org/uri/406 PDF summary of pharmacokinetics and efficacy]. This is being folded into the sections below, then supplemented.
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The authors made the following '''summary''' of their work on the triazolopyrazine series:
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-
"''We have made compounds in this series down to 16nM potency. The series also seems to have good in vitro HLM and hHEP stability Clint < 8.1 is compatible with 10nM potency. However, RLM remains stubbornly high, particularly for the more potent analogues translating to short half-lives in rat PK. The series also appears to have little polypharmacology or cytotoxicity. The project has so far not challenged the hypothesis that rat metabolism may not be a great model for human metabolism for this series.''"
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The aim of the campaign at the outset was to improve the metabolic stability and the pharmacokinetic properties of this series in rat so as to meet the once-dosing criteria (TCP1) set by MMV. New chemistry directed towards blocking the putative metabolic sites was a major part of the research prior to the data being contributed to OSM.  
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The authors also flagged some '''concerns''' regarding the series:
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==Sources of Data on the TP Series==
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"''Although [http://en.wikipedia.org/wiki/Dofetilide# dofetilide] binding looks weak or nil, the series has shown activity in a patch clamp assay at Essen (1-10uM) which is quite potent though with a window of >100 fold over Pfal potency. The series shows activity in Kieran Kirk’s PfATP4 assay which goes away for Pfal inactives in the series. In Kip Guy’s resistant mutants the picture is perhaps more mixed, but there is still support for the idea that some members of the series are weaker in the resistant strains. The series has no or weak >>1uM activity against gametocytes, no activity against Winzeler’s Pb liver stage and may have weak activity against ookinetes but the dose-response data has not been completed."''
+
[http://malaria.ourexperiment.org/uri/404# Initial briefing document] (A minor error in the briefing document referring to the amides has been [http://malaria.ourexperiment.org/uri/42d clarified].) Briefing document mostly folded into the below.<br>
 +
[http://malaria.ourexperiment.org/uri/406 PDF summary of pharmacokinetics and efficacy]. Needs folding into the below.<br>
 +
[http://malaria.ourexperiment.org/triazolopyrazine_se The current synthetic lab notebook]<br>
 +
[http://malaria.ourexperiment.org/osdd_malaria_shared/8106/MMV_triazolopyrazine_data.html Raw Data Spreadsheet on Compounds Prior to Donation to OSM] including available quantities of original samples<br>
 +
[http://malaria.ourexperiment.org/osdd_malaria_shared/8292/Collected_Amides_in_the_TP_Series_Triazolopyrazines_Series_4.html Summary of Data on Amides in the Series]<br>
 +
[http://malaria.ourexperiment.org/data/files/7384/FINAL%20REPORT%20MMV670652.doc Document from CRO describing synthetic chemistry of some members of series]<br>
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Possible next steps were also suggested for the triazolopyrazine series:
+
==Current Aims==
-
"''The series has good potency and" in vivo "efficacy with few toxicity concerns. The biggest issue is metabolic stability, as measured in rat in particular. Some possible future directions include:
+
1. Lead optimisation, to improve solubility and metabolic stability while maintaining potency.<br>
-
• Small scale changes around the side chains to attempt to balance potency and metabolism.
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2. Validation of PfATP4 activity as mechanism of action.<br>
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• Incorporation of a basic centre to increase volume as a potential fix for half-life. However, this might come at the expense of plasma concentration so would require high potency. In addition of the 29 compounds with a basic centre only one has a measured potency < 100nM.
+
-
• More significant structural changes. Of the changes made to the basic skeleton, the most successful might be the recent evaluation of the substitution position changes (eg MMV670945), possibly in combination with modifying the disposition of the N atoms in the core. Related compounds have been made by others and it would be wise to incorporate the learnings from these series into any plans to explore this substitution pattern further. The first few compounds look similar in terms of metabolic stability.''''
+
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More contents will gradually become part of this wiki (below) as the project progresses.
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===Current/Recent Activity===
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The raw data behind the briefing document will be released when it is obtained. New ion regulation assay data has already been received (below).
+
Last updated: Apr 4th 2014
-
===What Can the Community Do Now?===
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1. '''Which compounds to make next?''' - see [http://malaria.ourexperiment.org/osddmalaria_meeting_/9470 the most recent meeting] for links to the discussions on these points. Compounds in the [http://malaria.ourexperiment.org/the_osm_blog/9211 ether] and [http://malaria.ourexperiment.org/the_osm_blog/9519 amide] sub-series are being made. [https://github.com/OpenSourceMalaria/OSM_To_Do_List/issues?labels=Being+Synthesised+Now&page=1&state=open Automatically-updated list of compounds being made now] - some compounds may be from older series.
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Those wishing to contribute to OSM Series 4 should in the first instance read the [http://malaria.ourexperiment.org/uri/404# briefing document]. The first question is: Which 10 compounds ought to be made in Sept/Oct 2013 for evaluation?
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2. '''Does anyone possess compounds that could be relevant to this series already?''' It [http://cdsouthan.blogspot.se/2013/12/searching-triazolopyrazines-for-open.html currently appears] as though the answer is "no".
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Resources Needed: Chemists to either make new molecules, or help obtain existing compounds that might be relevant to this series.
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3. '''Resources needed''': a) Chemists to make molecules, b) Experienced medicinal chemists to digest the data on this wiki and suggest new compounds to make. You can contact the OSM Consortium in many ways (go to the "Join the Team" page [http://opensourcemalaria.org/ here].
-
===Current Aims===
+
==Notable Points about the TP Series==
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1. Hit-to-lead synthesis and biological evaluation, to improve potency and drug likeness.<br>
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* Compounds in this series have been identified down to 16 nM potency.
-
2. Validation of PfATP4 activity.<br>
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* Seems to have good ''in vitro'' HLM and hHEP stability Clint < 8.1 is compatible with 10 nM potency.
 +
* RLM remains stubbornly high, particularly for the more potent analogues translating to short half-lives in rat PK.
 +
* Series appears to have little polypharmacology or cytotoxicity.
 +
* Not so far challenged the hypothesis that rat metabolism may not be a great model for human metabolism for this series.
 +
* The series shows activity in Kieran Kirk’s PfATP4 assay which goes away for Pfal inactives in the series.
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===Current Questions for the Community===
+
==Concerns about the TP Series==
-
Starting-point questions are listed at the end of the [http://malaria.ourexperiment.org/uri/404# briefing document]. These translate into the initial question of '''Which 10 compounds ought to be made first?''' (also asked [https://github.com/OpenSourceMalaria/OSM_To_Do_List/issues/86 here] and '''Does anyone possess compounds that could be relevant to this series already?''' (asked [https://github.com/OpenSourceMalaria/OSM_To_Do_List/issues/87 here] and there is a separate section for results below).
+
* Although [http://en.wikipedia.org/wiki/Dofetilide# dofetilide] binding looks weak or nil, the series has shown activity in a patch clamp assay at Essen (1-10 uM) which is quite potent though with a window of >100 fold over Pfal potency.
 +
* In Kip Guy’s resistant mutants the picture is mixed, but there is still support for the idea that some members of the series are weaker in the resistant strains. The series has no or weak >>1uM activity against gametocytes, no activity against Winzeler’s Pb liver stage and may have weak activity against ookinetes but the dose-response data has not been completed.
-
==Prior Knowledge of the Series==
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==Project Strands of Current Interest==
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===Potency of Hits and Analogs===
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The biggest issue is metabolic stability, as measured in rat in particular. There are few toxicity concerns. Thus possible future directions:
 +
* Small scale changes around the side chains, particularly phenethyl to attempt to balance potency and metabolism. Other possibilities: a) N is tolerated in the ring, hasn’t been explored much recently. b) Is 3,4-diF the best substitution pattern? c) Some evidence (eg MMV669848) that the phenethyl side chain can be rigidified, perhaps the iso-indoline of that compound could be improved on with other ring systems and by more optimal substitution of the aromatic benzene ring of the isoindoline. d) The amide MMV670944 is interesting and shows good RLM stability, but many other amides failed to match its potency.
 +
* Incorporation of a basic centre to increase volume as a potential fix for half-life. However, this might come at the expense of plasma concentration so would require high potency. Of the 29 compounds with a basic centre only one (MMV670437, below) has a measured potency < 100 nM (actually 44 nM).
-
(Detail will be placed here, but for the moment please browse the [http://malaria.ourexperiment.org/uri/404# briefing document]).
+
[[Image:MMV670437.png|thumb|center|200px|MMV670437]]
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===Possible PfATP4 Activity Deduced from Parasite Ion Regulation Assays===
+
* More significant structural changes. Of the changes made to the basic skeleton, the most successful might be the recent evaluation of the substitution position changes (''e.g.,'' MMV670945), possibly in combination with modifying the disposition of the N atoms in the core. Related compounds have been made by others and it would be wise to incorporate the learnings from these series into any plans to explore this substitution pattern further. The first few compounds look similar in terms of metabolic stability.
 +
 
 +
==SAR==
 +
 
 +
===Cycloaliphatic Triazole Substituent===
 +
 
 +
Attempts at lowering the lipophilicity of the TP compounds by replacing the triazole aryl substituent with a cyclo(hetero)aliphatic group, linked either by the heteroatom or otherwise (e.g. piperidine, tetrahydropyran, indoline or isoindoline) lowered the potency against PfNF54, as did an aniline substituent.
 +
 
 +
[[Image:Cycloaliphatic Triazole Subst.png|thumb|center|200px|Changes to the Triazole Substitution]]
 +
 
 +
===Core Modification of Pyrazine Ring===
 +
 
 +
Based on an assumption that the pyrazine moiety of the TP could undergo AO metabolism at positions alpha- to the nitrogen a few compounds were made with different R groups (Cl, Me, NH<sub>2</sub>, NEt<sub>2</sub>). However, all these compounds lost potency against PfNF54.
 +
 
 +
[[Image:Pyrazine Subst.png|thumb|center|200px|Changes to the Pyrazine Substitution]]
 +
 
 +
===Core Modification of Triazole Ring===
 +
 
 +
Two compounds based on imadazopyrazines were made (MMV669846 and MMV670250, below). Both showed reduced potency against PfNF54 ''vs.'' the corresponding TP compound. The RLM stability of one was found to be poor. Approx 20 structures were made with variations to the 6,5 core system. MMV669846 was the most potent. As most of the analogues were >1 μM potency, fewer were tested in RLM (quite a few in HLM). Of the 4 tested in RLM, the greatest stability had a Clint of 109, (HLM 9.5), several had HLM Clint 8 or less, particularly after moving or removing the N from the pyrazine ring.
 +
 
 +
[[Image:Core Mod of Triazole.png|thumb|center|600px|Core Modification of Triazine Ring]]
 +
 
 +
''(Note - original briefing document contained two entries for MMV669846 at this point with different potencies - need check of original spreadsheet, to verify the above is correct)''
 +
 
 +
===Transposing the Pyrazine side-chain===
 +
 
 +
The side-chain on the pyrazine ring was shifted to the adjacent carbon. The chain length was varied (n= 0,1,2) and linked through either O or N.
 +
 
 +
[[Image:Wiki Shifted Side Chain.png|thumb|center|200px|Shifted Side Chain]]
 +
 
 +
Among the ethers, the phenethyl ether (X = O, n =2) showed good potency (Pfal IC50 33 nM) but a poor stability in RLM (Cl 100 mL/min/Kg)/
 +
 
 +
[[Image:Wiki Shifted Ethers and Amines.png|thumb|center|600px|Shifted Ethers and Amines]]
 +
 
 +
[[Image:Pyrazine Side Chain.png|thumb|center|400px]]
 +
 
 +
===Pyrazine side-chain modifications - non amides===
 +
 
 +
The phenethyl ether side-chain on the pyrazine ring was flagged as a metabolic hot-spot and so several strategies were adopted to mitigate this risk.  The following findings were observed:
 +
* Changing the length of the side-chain to anything other than 3 atoms severely lowers potency against PfNF54
 +
* The linker atom to the pyrazine ring is crucial (O>>C>N)
 +
* Heteroatoms in the side-chain lowered potency with the exception of MMV669848 (featuring an isoindolino-methyl group on the pyrazine ring) although this compound had poor RLM stability.
 +
* Constraining the linear side-chain into ring systems (e.g, azetidines, pyrrolidines, pyrazoles) severely reduced potency.
 +
 
 +
[[Image:MMV669848.png|thumb|center|200px|MMV669848]]
 +
 
 +
A 2-naphthol substituent on the pyrazine ring showed reasonable potency against Pfal (IC50 114 nM) but suffered from poor RLM stability. Several hetero-analogs of 2-naphthol e.g. indole, indazole, quinoline, chroman, benzisoxazole, quinazoline, etc were made but all lost potency against the parasite.
 +
 
 +
The phenyethyl chain was replaced by an aromatic group in an attempt to mitigate the potential metabolism of the ethyl chain. Several compounds with a phenol substituent on the pyrazine ring were made. Some substituted phenolates were metabolically more stable '''in vitro''' as well as ''in vivo'' in rat, although with reduced potency against Pfal.
 +
 
 +
[[Image:MMV669784.png|thumb|center|400px|MMV669784]]
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 +
Considering that the benzylic position in the phenethyl side-chain is prone to metabolic oxidation, several compounds having mono- and di-substitution in the benzylic position were made. Di-Substitution lowered the potency considerably whereas mono-substitution with OMe, OCHF<sub>2</sub>, CH<sub>2</sub>OH, NMe<sub>2</sub> groups retained good potency. Additional substitution alpha- to the ether oxygen led to complete loss of potency. The alpha-OCHF<sub>2</sub> compound MMV670652, with a p-CN-phenyl group on the triazole ring, showed better RLM stability (cLogP effect).
 +
 
 +
===Pyrazine side-chain modifications - amides===
 +
 
 +
A small library of amides (including the ''m''-Cl benzylamide, MMV668958) showed promising potency. However, other amides (derived from aliphatic or anilines) either showed lower potency or were inactive. The RLM of MMV668958 was poor - perhaps due to benzylic oxidation. Alpha-substitution at the benzylic position or constraining the benzylamine into an aminoindane did not improve potency. Several attempts to make aniline-amides with improved potency against Pfal failed. Loading the aniline ring with lipophilic substituents marginally improved potency but led to poor RLM stability.
 +
 
 +
[[Image:Amide Library.png|thumb|center|900px|Amides Synthesised and Tested pre-OSM]]
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 +
The ''p''-Cl benzanilide MMV670246, although not active against PfNF54, showed good RLM stability perhaps due to lack of benzylic metabolism. However, its rat PK showed high clearance. The potent ''m''-Cl (MMV669542) analog had poor RLM stability.
 +
 
 +
[[Image:MMV670246.png|thumb|center|400px|Further PK Details on MMV670246]]
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 +
==Pharmacokinetics==
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 +
Only one compound in this Series first made at the CRO was measured in rat PK and that was the relatively weak amide MMV670246. The curve is shown for oral & IV legs & parameters are below.
 +
 
 +
[[Image:MMV670246_rat_PK.png|centre|MMV670246 in rat PK|thumb|400px]]
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 +
[[http://openwetware.org/wiki/Image:MMV670246_rat_PK_data.png Data for MMV670246 in rat PK]]
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 +
One of the compounds with better ''in vitro'' balance is MMV670652 with potency at 17 nM, HLM Clint < 8 ul/min/mg and RLM Clint at 30 ul/min/mg. This compound has not been in rat PK. Additionally it may be possible to improve potency by synthesis of the more potent enantiomer.
 +
 
 +
[[ Image:MMV670562.png|thumb| centre |300px|MMV670652 ]]
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A [http://malaria.ourexperiment.org/uri/406 PDF summary of pharmacokinetics and efficacy] provided as an initial source contained efficacy information for the compound MMV639565.
 +
 
 +
[[ Image:MMV639565.png|thumb| centre |400px|MMV639565 ]]
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The therapeutic efficacy of MMV639565 against P falciparum growing in peripheral blood of NODscidIL2Rγnull mice engrafted with human erythrocytes (po dosing, qd for 4 days) is shown below.  A rapid parasite clearance - ED<sub>90</sub> 6.3 mg/kg was recorded.
 +
 
 +
[[ Image:MMV639565efficacy.png|thumb| centre |500px|MMV639565 Therapeutic Efficacy ]]
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 +
Values in brackets indicate dose corrected according to quality control of formulation.
 +
 
 +
==Other Observations==
 +
 
 +
As would be expected HLM ''vs.'' RLM shows a general correlation with approx 4-fold shift on average. However, for most of the more potent analogs, this increases to over 10-fold. The figure below shows the 4 sub 30nM compounds with HLM & RLM measured.
 +
 
 +
[[Image:HLM_vs_RLM.png|thumb|400px|HLM vs RLM|centre]]
 +
 
 +
Lipophilicity & lipophilic efficiency:
 +
 
 +
Few compounds achieve a lipophilic efficiency (as measured by pIC50 – AlogP) of greater than 4.0
 +
 
 +
[[Image:Lipophilicity_&_lipophilic_efficiency.png|centre|400px|thumb|Lipophilic efficiency]]
 +
 
 +
==Potential next steps - suggested in original pre-OSM briefing document==
 +
 
 +
The series has good potency and ''in vivo'' efficacy with few toxicity concerns. The biggest issue is metabolic stability, as measured in rat in particular. Some possible future directions include:
 +
* Small scale changes around the side chains, particularly phenethyl to attempt to balance potency and metabolism
 +
* N is tolerated in the ring, hasn’t been explored much recently
 +
* Is 3,4-diF the best substitution pattern ?
 +
* Some evidence (eg. MMV669848) that the phenethyl side chain can be rigidified, perhaps the iso-indoline of that compound could be improved on with other ring  systems and by more optimal substitution of the aromatic benzene ring of the isoindoline.
 +
* The amide MMV670944 is interesting and shows good RLM stability, but many other amides failed to match its potency
 +
* Incorporation of a basic centre to increase volume as a potential fix for half-life. However, this might come at the expense of plasma concentration so would require high potency. In addition of the 29 compounds with a basic centre only one has a measured potency < 100nM.
 +
 
 +
[[Image:MMV670437.png|centre|thumb|200px|MMV670437]]
 +
 
 +
* More significant structural changes. Of the changes made to the basic skeleton, the most successful might be the recent evaluation of the substitution position changes (eg MMV670945), possibly in combination with modifying the disposition of the N atoms in the core. Related compounds have been made by others and it would be wise to incorporate the learnings from these series into any plans to explore this substitution pattern further. The first few compounds look similar in terms of metabolic stability.
 +
 
 +
==Possible PfATP4 Activity Deduced from Parasite Ion Regulation Assays==
The following five compounds were evaluated in parasite ion regulation assays in the [http://biology.anu.edu.au/kiaran_kirk/ Kirk Laboratory]; the hypothesis is that PfATP4 is a Na<sup>+</sup> ATPase that exports Na<sup>+</sup> and imports H<sup>+</sup> (or equivalent) and that the effects of the compounds on Na<sup>+</sup> concentration and pH are attributable to inhibition of this activity. Structures, potency, metabolism/solubility and raw PfATP4 assay data are [http://malaria.ourexperiment.org/biological_data/7934/ here].
The following five compounds were evaluated in parasite ion regulation assays in the [http://biology.anu.edu.au/kiaran_kirk/ Kirk Laboratory]; the hypothesis is that PfATP4 is a Na<sup>+</sup> ATPase that exports Na<sup>+</sup> and imports H<sup>+</sup> (or equivalent) and that the effects of the compounds on Na<sup>+</sup> concentration and pH are attributable to inhibition of this activity. Structures, potency, metabolism/solubility and raw PfATP4 assay data are [http://malaria.ourexperiment.org/biological_data/7934/ here].
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(i.e. note the correlation: compound inactive in these assays is the inactive analog in the plasmodium screen)
(i.e. note the correlation: compound inactive in these assays is the inactive analog in the plasmodium screen)
-
==Other Sources of Compounds in this Series==
+
==Physicochemical/metabolism==
-
([https://github.com/OpenSourceMalaria/OSM_To_Do_List/issues/87 Community Request])
+
The following compounds were [http://malaria.ourexperiment.org/uri/457 sent] (associated [https://github.com/OpenSourceMalaria/OSM_To_Do_List/issues/107 discussion]) and [http://malaria.ourexperiment.org/uri/490 data were obtained].
 +
 
 +
[[Image:Physicochem and sol compounds.png|thumb|center|600px|Compounds Evaluated for Solubility and Metabolic Clearance]]
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 +
Solubility is low and improvement should be one of the goals for lead optimisation. The metabolic stability is not a mouse specific event and needs to be improved to deliver a candidate. Met ID data may help in the design of new analogs with improved stability, but we could also look at correlations with Log D etc. to drive new target design)
 +
 
 +
===Metabolism ID===
 +
 
 +
The following compounds were [http://malaria.ourexperiment.org/uri/457 sent] and [http://malaria.ourexperiment.org/uri/49a data were obtained].
 +
 
 +
[[Image:Compounds sent for MetID.png|thumb|center|600px|Compounds Evaluated for Metabolic ID]]
 +
 
 +
The data suggest that the core heterocyclic ring is subjected to an oxidation (though it is not clear the extent to which the compound is oxidized, only that certain peaks are present). The nature of the side chain appeared to have little influence on the results, suggesting alteration of the side chain is probably not the way to stop turnover, meaning it is likely that amides made in this series in the near future will also suffer from this problem. Nevertheless the efficacy was seen as sufficiently promising that amides remain attractive as they are. i.e. they are sufficiently robust for this point in the project. Residual question: what is the mechanism of the clearance (oxidation)? Is it CYP mediated? Open action item [https://github.com/OpenSourceMalaria/OSM_To_Do_List/issues/124 here].
 +
 
 +
==hERG Data==
 +
 
 +
MMV669844 and MMV670944 ([http://malaria.ourexperiment.org/osm_procedures/9557/Preparation_of_OSMS175.html OSM-S-175]) were evaluated in a hERG assay, with [http://malaria.ourexperiment.org/biological_data/9562 the data] indicating issues with both compounds that will need to be addressed.
 +
 
 +
==Synthetic Chemistry==
 +
 
 +
===Synthetic Design===
 +
 
 +
The approach to the synthesis of Series 4 up until early 2014 has involved the functionalisation of an existing pyrazine core, followed by a cylization reaction to give the triazolopyrazine.
 +
 
 +
Literature examples of cyclization to give triazolopyrazine (Tom to insert one example).
 +
 
 +
Synthetic chemistry for OSM in Jan/Feb 2014 has been focussed on two main targets in Series 4 (the triazolopyrazines) - the [http://malaria.ourexperiment.org/the_osm_blog/8028 ether linked compounds and the amide compounds], shown below along with specific examples of known, potent compounds in each series.
 +
 
 +
[[Image:Example Targets.png|thumb|center|250px|Examples of ether and amide target molecules.]]
 +
 
 +
===Synthesis of the Ether-Linked Series===
 +
The ether series contains many active compounds. MMV670652 shown above was evaluated as a racemate, and Alice has been attempting a resynthesis with a view to the separation of enantiomers to evaluate the properties of the active enantiomer. [https://github.com/OpenSourceMalaria/OSM_To_Do_List/issues/115 Here's the original issue] for the synthesis of compound MMV670652, as well as the [https://github.com/OpenSourceMalaria/OSM_To_Do_List/issues/111 first discussion of how we might separate the enantiomers], and a [https://github.com/OpenSourceMalaria/OSM_To_Do_List/issues/120 newer issue] created after a recent meeting. One of the complexities of the synthesis is the [https://github.com/OpenSourceMalaria/OSM_To_Do_List/issues/119 difluoromethylation step] (original summary of problem is [http://malaria.ourexperiment.org/the_osm_blog/8470 here]). It's not clear that this group is needed - there are [http://malaria.ourexperiment.org/the_osm_blog/8550 related structures that could be tried], and [http://malaria.ourexperiment.org/biological_data/9191 compounds lacking this group (and indeed the whole stereogenic centre) are still active].
 +
 
 +
A key synthetic stumbling block has been the union of the two halves of the molecule - an alcohol ('''2''') and the heterocycle chloride ('''7'''), for example in [http://malaria.ourexperiment.org/triazolopyrazine_se/9073 this recent reaction]. The reactions were for a long time not proceeding cleanly, and this was also found by the CRO that first did it - report was posted [http://malaria.ourexperiment.org/data/files/9856/CRO%20Synth%20Series%204.doc here]. Jo Ubels [http://malaria.ourexperiment.org/the_osm_blog/9357 performed a lit survey] and found conditions involving a crown ether that [http://malaria.ourexperiment.org/triazolopyrazine_se/9459 now appear to provide the desired ether]. The synthesis of the chiral alcohol fragment needed specifically for MMV670652, via '''1''', [http://malaria.ourexperiment.org/triazolopyrazine_se/8378 requires a source of cyanide], and we're currently out of stock in Sydney. Thus currently simpler alcohols (i.e. ArCH<sub>2</sub>CH<sub>2</sub>OH) are being used in this step. A [https://github.com/OpenSourceMalaria/OSM_To_Do_List/issues/167 new enantioselective approach] is also being developed by Alice in the meantime.
 +
 
 +
[[Image:Route_to_Ether_Linked.png|thumb|center|500px|Synthetic route to ether.]]
 +
 
 +
Another synthetic challenge being addressed is the efficiency of the cyclization to give 7 from the precursor 6. [http://malaria.ourexperiment.org/triazolopyrazine_se/8217 This has been achieved], but not in the ca. 50% yields reported by the original CRO. A new Honours student in the group, [http://malaria.ourexperiment.org/the_osm_blog/8894 Jo Ubels], is now looking at this step to work out what is going on and how we can make this reaction more efficient and this has been the subject of research by an Edinburgh student working with Patrick Thomson, [http://malaria.ourexperiment.org/triazolopyrazine_se/8335 Devon Scott]. There appear to be significant issues to do with solubility of the material throughout the reaction. The [http://malaria.ourexperiment.org/triazolopyrazine_se/8967 synthesis of the precursor 7] is straightforward.
 +
 
 +
===Synthesis of the Amide-Linked Series===
 +
 
 +
The amide series is interesting because there are [http://openwetware.org/wiki/OpenSourceMalaria:Triazolopyrazine_%28TP%29_Series#Pyrazine_side-chain_modifications_-_amides potent compounds] with such a structure that appear to have desirable PK properties, but potency needs to be enhanced. Following an analysis of commercially-available [http://malaria.ourexperiment.org/osm_logos_and_templ/8578 primary] and [http://malaria.ourexperiment.org/the_osm_blog/8936 secondary] amines that could be incorporated, the [https://github.com/OpenSourceMalaria/OSM_To_Do_List/issues/123 likely physical properties of the resulting compounds] as well as an analysis of [https://github.com/OpenSourceMalaria/OSM_To_Do_List/issues/151 those amines we had to hand] in the building, synthesis was started (scheme below). It has been noted that an efficient approach here would be to use the same intermediate as for the ether series ('''7''', above) and invent a new carbonylation method ([https://github.com/OpenSourceMalaria/OSM_To_Do_List/issues/126 original issue], [http://malaria.ourexperiment.org/the_osm_blog/8627 Inga's lit survey], [http://malaria.ourexperiment.org/the_osm_blog/8480 Alice's original appeal for a collaborator] and a [http://malaria.ourexperiment.org/the_osm_blog/8173 summary of the associated discussion].
 +
 
 +
For the moment, however, [https://github.com/OpenSourceMalaria/OSM_To_Do_List/issues/121 it was decided] to pursue an [https://github.com/OpenSourceMalaria/OSM_To_Do_List/issues/101 alternative route] wherein the extra carbon atom is incorporated from the start using acid '''9'''. This molecule is commercially available but is expensive, but we now have routes to it in the lab. [http://downwithmalariaupwithopensource.blogspot.co.uk/2013/10/direction-and-strategy-for-bsc-projects.html Patrick] and [https://github.com/OpenSourceMalaria/OSM_To_Do_List/issues/150 Inga] planned the synthesis from the pyrazine acid and then [http://malaria.ourexperiment.org/triazolopyrazine_se/9147 Inga completed] a synthesis of it, as has [http://malaria.ourexperiment.org/triazolopyrazine_se/8899 Eduvie using the same approach] (though data are missing for these latter experiments). Sabin has [https://github.com/OpenSourceMalaria/OSM_To_Do_List/issues/146 successfully demonstrated] his very nice alternative route to this compound in one step from the methyl precursor ([http://malaria.ourexperiment.org/triazolopyrazine_se/8929 data]). [http://malaria.ourexperiment.org/triazolopyrazine_se/9189 Tom is now scaling up] the Patrick/Eduvie/Inga approach to bring through more material.
 +
 
 +
For the subsequent coupling of the pyrazine acid with an amine, Tom and Alice have identified a [http://malaria.ourexperiment.org/triazolopyrazine_se/9142 clean coupling reaction to give the amide '''10'''] using T3P that worked much better than [http://malaria.ourexperiment.org/triazolopyrazine_se/8937 an attempt to go via the acid chloride], and Alice has seen [http://malaria.ourexperiment.org/triazolopyrazine_se/9167 excellent preliminary results] in the coupling of a related amide with hydrazine, condensation with aldehydes (this chemistry has been [http://malaria.ourexperiment.org/the_osm_blog/9169 surveyed by Inga]) and is now attempting the crucial cyclization reaction to give what would be the final amide. Should this work, we would then have access to a wide range of new members in the amide series. [https://github.com/OpenSourceMalaria/OSM_To_Do_List/issues/152 Here] is one of Tom's current "Being Synthesised Now" Github posts in this series. [https://github.com/OpenSourceMalaria/OSM_To_Do_List/issues/157 Here] is one of Alice's where she is altering the northeast part of the molecule. There was a suggestion that hydrazone isomers may be present and may be differently reactive towards the oxidative cyclisation conditions (GH issue [https://github.com/OpenSourceMalaria/OSM_To_Do_List/issues/97 97]).
 +
 
 +
[[Image:Route_to_Amide_Linked.png|thumb|center|500px|Synthetic route to amide.]]
 +
 
 +
==Chirality/Stereogenic Centres in This Series==
 +
 
 +
MMV670652 shown above was evaluated as a racemate, and there was interest in separating enantiomers: [https://github.com/OpenSourceMalaria/OSM_To_Do_List/issues/115 original issue] then [https://github.com/OpenSourceMalaria/OSM_To_Do_List/issues/111 discussion of how we might separate the enantiomers], and a [https://github.com/OpenSourceMalaria/OSM_To_Do_List/issues/120 more recent issue]. It is not clear that this group is needed - there are [http://malaria.ourexperiment.org/the_osm_blog/8550 related structures that could be tried], and [http://malaria.ourexperiment.org/biological_data/9191 compounds lacking this group (and indeed the whole stereogenic centre) are still active]. MMV669844 (CH3 rather than CHF2) was synthesised using a Sharpless epoxidation on the precursor styrene, but there are no data on the enantiomeric excess of the product - it is shown as a single enantiomer in the original briefing document.
 +
 
 +
==Other Sources of Compounds Relevant to this Series==
 +
 
 +
([https://github.com/OpenSourceMalaria/OSM_To_Do_List/issues/87 Original community request])<br>
 +
The compound series [http://cdsouthan.blogspot.se/2013/12/searching-triazolopyrazines-for-open.html appears to be novel/unusual] with little similarity to compounds in PubChem/Surechem, though Chris Southan (in the previous link) found a related compound ([https://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=7118230 CID 7118230]) with [https://pubchem.ncbi.nlm.nih.gov/assay/assay.cgi?aid=504832 reported antimalarial activity], with a structure similar to those described above for modification of the triazolopyrazine core. Jo Ubels also [http://malaria.ourexperiment.org/the_osm_blog/8894/An_Introduction__Joanna.html carried out a search] and found few compounds similar to Series 4.<br>
 +
[https://github.com/OpenSourceMalaria/OSM_To_Do_List/issues/147 A similarity was noted] between Series 4 compounds and a compound from Novartis likely active against vivax. Paul Willis suggested the mechanism of action of the Novartis compounds was likely PI4K - data and reasoning given in [http://malaria.ourexperiment.org/triazolopyrazine_se/8912 this lab book entry]. Chris Southan [http://cdsouthan.blogspot.se/2013/12/searching-triazolopyrazines-for-open.html modelled the OSM and Novartis compounds], which provides additional evidence for a different mechanism of action.
==Strings for Google==
==Strings for Google==
Line 89: Line 283:
-
'''MMV669000''': O=C(N1CC(C=CC=C2)=C2C1)C3=CN=CC4=NN=C(C5=CC=C(OC(F)F)C=C5)N43; InChI=1S/C21H15F2N5O2/c22-21(23)30-16-7-5-13(6-8-16)19-26-25-18-10-24-9-17(28(18)19)20(29)27-11-14-3-1-2-4-15(14)12-27/h1-10,21H,11-12H2<br>
+
'''MMV668955''' FC(F)OC(C=C1)=CC=C1C2=NN=C3N2C(CCO)CNC3; InChI=1S/C14H16F2N4O2/c15-14(16)22-11-3-1-9(2-4-11)13-19-18-12-8-17-7-10(5-6-21)20(12)13/h1-4,10,14,17,21H,5-8H2<br>
-
'''MMV669304''': FC(F)OC(C=C1)=CC=C1C2=NN=C3C=NC=C(CCCC4=CC=CC=C4)N32; InChI=1S/C21H18F2N4O/c22-21(23)28-18-11-9-16(10-12-18)20-26-25-19-14-24-13-17(27(19)20)8-4-7-15-5-2-1-3-6-15/h1-3,5-6,9-14,21H,4,7-8H2<br>
+
'''MMV668958''' FC(F)OC(C=C1)=CC=C1C2=NN=C3N2C(C(NCC4=CC(Cl)=CC=C4)=O)=CN=C3; InChI=1S/C20H14ClF2N5O2/c21-14-3-1-2-12(8-14)9-25-19(29)16-10-24-11-17-26-27-18(28(16)17)13-4-6-15(7-5-13)30-20(22)23/h1-8,10-11,20H,9H2,(H,25,29)<br>
-
'''MMV669360''': FC(F)OC(C=C1)=CC=C1C2=NN=C3C=NC=C(COCC4=CC=C(F)C(F)=C4)N32; InChI=1S/C20H14F4N4O2/c21-16-6-1-12(7-17(16)22)10-29-11-14-8-25-9-18-26-27-19(28(14)18)13-2-4-15(5-3-13)30-20(23)24/h1-9,20H,10-11H2<br>
+
'''MMV668960''' FC(F)OC(C=C1)=CC=C1C2=NN=C3N2CCNC3; InChI=1S/C12H12F2N4O/c13-12(14)19-9-3-1-8(2-4-9)11-17-16-10-7-15-5-6-18(10)11/h1-4,12,15H,5-7H2<br>
-
'''MMV669542''': FC(F)OC(C=C1)=CC=C1C2=NN=C3C=NC=C(C(NC4=CC=CC(Cl)=C4)=O)N32; InChI=1S/C19H12ClF2N5O2/c20-12-2-1-3-13(8-12)24-18(28)15-9-23-10-16-25-26-17(27(15)16)11-4-6-14(7-5-11)29-19(21)22/h1-10,19H,(H,24,28)<br>
+
'''MMV668962''' FC(F)OC(C=C1)=CC=C1C2=NN=C3N2CCN(C(CN)=O)C3; InChI=1S/C14H15F2N5O2/c15-14(16)23-10-3-1-9(2-4-10)13-19-18-11-8-20(12(22)7-17)5-6-21(11)13/h1-4,14H,5-8,17H2<br>
-
'''MMV669848''': FC(F)OC(C=C1)=CC=C1C2=NN=C3C=NC=C(CN4CC(C=CC=C5)=C5C4)N32; InChI=1S/C21H17F2N5O/c22-21(23)29-18-7-5-14(6-8-18)20-26-25-19-10-24-9-17(28(19)20)13-27-11-15-3-1-2-4-16(15)12-27/h1-10,21H,11-13H2
+
'''MMV669000''' O=C(N1CC(C=CC=C2)=C2C1)C3=CN=CC4=NN=C(C5=CC=C(OC(F)F)C=C5)N43; InChI=1S/C21H15F2N5O2/c22-21(23)30-16-7-5-13(6-8-16)19-26-25-18-10-24-9-17(28(18)19)20(29)27-11-14-3-1-2-4-15(14)12-27/h1-10,21H,11-12H2<br>
 +
'''MMV669025''' FC1=C(F)C=CC(CCOC2=CNC(C3=NN=C(C4=CC=C(OC(F)F)C=C4)N32)=O)=C1; InChI=1S/C20H14F4N4O3/c21-14-6-1-11(9-15(14)22)7-8-30-16-10-25-19(29)18-27-26-17(28(16)18)12-2-4-13(5-3-12)31-20(23)24/h1-6,9-10,20H,7-8H2,(H,25,29)<br>
 +
'''MMV669304''' FC(F)OC(C=C1)=CC=C1C2=NN=C3C=NC=C(CCCC4=CC=CC=C4)N32; InChI=1S/C21H18F2N4O/c22-21(23)28-18-11-9-16(10-12-18)20-26-25-19-14-24-13-17(27(19)20)8-4-7-15-5-2-1-3-6-15/h1-3,5-6,9-14,21H,4,7-8H2<br>
 +
'''MMV669310''' FC(F)OC(C=C1)=CC=C1C2=NN=C3N2C(CCCC4=CC=CC=C4)CNC3; InChI=1S/C21H22F2N4O/c22-21(23)28-18-11-9-16(10-12-18)20-26-25-19-14-24-13-17(27(19)20)8-4-7-15-5-2-1-3-6-15/h1-3,5-6,9-12,17,21,24H,4,7-8,13-14H2<br>
 +
'''MMV669360''' FC(F)OC(C=C1)=CC=C1C2=NN=C3C=NC=C(COCC4=CC=C(F)C(F)=C4)N32; InChI=1S/C20H14F4N4O2/c21-16-6-1-12(7-17(16)22)10-29-11-14-8-25-9-18-26-27-19(28(14)18)13-2-4-15(5-3-13)30-20(23)24/h1-9,20H,10-11H2<br>
 +
'''MMV669542''' FC(F)OC(C=C1)=CC=C1C2=NN=C3C=NC=C(C(NC4=CC=CC(Cl)=C4)=O)N32; InChI=1S/C19H12ClF2N5O2/c20-12-2-1-3-13(8-12)24-18(28)15-9-23-10-16-25-26-17(27(15)16)11-4-6-14(7-5-11)29-19(21)22/h1-10,19H,(H,24,28)<br>
 +
 
 +
'''MMV669844''' [C@H](COc1cncc2n1c(nn2)c1ccc(cc1)C#N)(c1cc(c(cc1)F)F)OC
 +
 
 +
'''MMV669846''' FC1=C(F)C=CC(CCOC2=CN=CC3=NC=C(C4=CC=C(Cl)C=C4)N32)=C1; InChI=1S/C20H14ClF2N3O/c21-15-4-2-14(3-5-15)18-10-25-19-11-24-12-20(26(18)19)27-8-7-13-1-6-16(22)17(23)9-13/h1-6,9-12H,7-8H2<br>
 +
'''MMV669848''' FC(F)OC(C=C1)=CC=C1C2=NN=C3C=NC=C(CN4CC(C=CC=C5)=C5C4)N32; InChI=1S/C21H17F2N5O/c22-21(23)29-18-7-5-14(6-8-18)20-26-25-19-10-24-9-17(28(19)20)13-27-11-15-3-1-2-4-16(15)12-27/h1-10,21H,11-13H2<br>
 +
'''MMV670246''' FC(F)OC(C=C1)=CC=C1C2=NN=C3N2C(C(NC4=CC=C(Cl)C=C4)=O)=CN=C3; InChI=1S/C19H12ClF2N5O2/c20-12-3-5-13(6-4-12)24-18(28)15-9-23-10-16-25-26-17(27(15)16)11-1-7-14(8-2-11)29-19(21)22/h1-10,19H,(H,24,28)<br>
 +
'''MMV670250''' FC1=C(F)C=CC(CCOC2=CN=CC3=CN=C(C4=CC=C(Cl)C=C4)N32)=C1; InChI=1S/C20H14ClF2N3O/c21-15-4-2-14(3-5-15)20-25-11-16-10-24-12-19(26(16)20)27-8-7-13-1-6-17(22)18(23)9-13/h1-6,9-12H,7-8H2<br>
 +
'''MMV670437''' FC(F)OC(C=C1)=CC=C1C2=NN=C3C=NC=C(OC[C@H](N(C)C)C4=CC(F)=C(F)C=C4)N32; InChI=1S/C22H19F4N5O2/c1-30(2)18(14-5-8-16(23)17(24)9-14)12-32-20-11-27-10-19-28-29-21(31(19)20)13-3-6-15(7-4-13)33-22(25)26/h3-11,18,22H,12H2,1-2H3/t18-/m0/s1<br>
 +
 
 +
'''MMV670936''' C(COc1cncc2n1c(nn2)c1cnc(cc1)C(F)(F)F)c1cc(c(cc1)F)F
 +
 
 +
'''MMV670944''' ([http://malaria.ourexperiment.org/osm_procedures/9557/Preparation_of_OSMS175.html OSM-S-175]) FC(F)OC(C=C1)=CC=C1C2=NN=C3N2C(C(NC4=CC=NC(F)=C4)=O)=CN=C3; InChI=1S/C18H11F3N6O2/c19-14-7-11(5-6-23-14)24-17(28)13-8-22-9-15-25-26-16(27(13)15)10-1-3-12(4-2-10)29-18(20)21/h1-9,18H,(H,23,24,28)<br>
 +
'''MMV670945''' FC(F)OC(C=C1)=CC=C1C2=NN=C3N2C=C(OCCC4=CC=C(F)C(F)=C4)N=C3; InChI=1S/C20H14F4N4O2/c21-15-6-1-12(9-16(15)22)7-8-29-18-11-28-17(10-25-18)26-27-19(28)13-2-4-14(5-3-13)30-20(23)24/h1-6,9-11,20H,7-8H2<br>
 +
'''MMV670946''' FC(F)OC(C=C1)=CC=C1C2=NN=C3N2C=C(OCC4=CC=C(F)C(F)=C4)N=C3; InChI=1S/C19H12F4N4O2/c20-14-6-1-11(7-15(14)21)10-28-17-9-27-16(8-24-17)25-26-18(27)12-2-4-13(5-3-12)29-19(22)23/h1-9,19H,10H2<br>
 +
'''MMV670949''' FC(F)OC(C=C1)=CC=C1C2=NN=C3N2C=C(NCC4=CC=C(F)C(F)=C4)N=C3; InChI=1S/C19H13F4N5O/c20-14-6-1-11(7-15(14)21)8-24-16-10-28-17(9-25-16)26-27-18(28)12-2-4-13(5-3-12)29-19(22)23/h1-7,9-10,19,24H,8H2<br>
 +
'''MMV671655''' FC(F)OC(C=C1)=CC=C1C2=NN=C3N2C=C(NC4=CC=C(F)C(F)=C4)N=C3; InChI=1S/C18H11F4N5O/c19-13-6-3-11(7-14(13)20)24-15-9-27-16(8-23-15)25-26-17(27)10-1-4-12(5-2-10)28-18(21)22/h1-9,18,24H<br>
 +
'''MMV671926''' FC(F)OC(C=C1)=CC=C1C2=NN=C3N2C=C(OC4=CC=C(F)C(F)=C4)N=C3; InChI=1S/C18H10F4N4O2/c19-13-6-5-12(7-14(13)20)27-16-9-26-15(8-23-16)24-25-17(26)10-1-3-11(4-2-10)28-18(21)22/h1-9,18H<br>
 +
'''MMV671927''' FC(F)OC(C=C1)=CC=C1C2=NN=C3N2C=C(NCCC4=CC=C(F)C(F)=C4)N=C3; InChI=1S/C20H15F4N5O/c21-15-6-1-12(9-16(15)22)7-8-25-17-11-29-18(10-26-17)27-28-19(29)13-2-4-14(5-3-13)30-20(23)24/h1-6,9-11,20,25H,7-8H2<br>
 +
'''MMV672619''' FC(F)OC(C=C1)=CC=C1C2=NN=C3N2C=C(O[C@@H]4C(N(C5CC5)[C@@H]4C6=CC(F)=C(F)C=C6)=O)N=C3; InChI=1S/C24H17F4N5O3/c25-16-8-3-13(9-17(16)26)20-21(23(34)33(20)14-4-5-14)36-19-11-32-18(10-29-19)30-31-22(32)12-1-6-15(7-2-12)35-24(27)28/h1-3,6-11,14,20-21,24H,4-5H2/t20-,21+/m1/s1<br>
 +
'''MMV672939''' FC(F)OC(C=C1)=CC=C1C2=NN=C3C=CC(OCCC4=CC(F)=C(F)C=C4)=NN32; InChI=1S/C20H14F4N4O2/c21-15-6-1-12(11-16(15)22)9-10-29-18-8-7-17-25-26-19(28(17)27-18)13-2-4-14(5-3-13)30-20(23)24/h1-8,11,20H,9-10H2<br>
 +
'''MMV672942''' FC(F)OC(C=C1)=CC=C1C2=NN=C3C=CC(NCCC4=CC=C(F)C=C4)=NN32; InChI=1S/C20H16F3N5O/c21-15-5-1-13(2-6-15)11-12-24-17-9-10-18-25-26-19(28(18)27-17)14-3-7-16(8-4-14)29-20(22)23/h1-10,20H,11-12H2,(H,24,27)<br>
 +
'''MMV672990''' FC(F)OC(C=C1)=CC=C1C2=NN=C3N2C=C(NC(CC4=CC=CC(Cl)=C4)=O)N=C3; InChI=1S/C20H14ClF2N5O2/c21-14-3-1-2-12(8-14)9-18(29)25-16-11-28-17(10-24-16)26-27-19(28)13-4-6-15(7-5-13)30-20(22)23/h1-8,10-11,20H,9H2,(H,25,29)<br>
 +
'''MMV672992''' N#CC(C=C1)=CC=C1C2=NN=C3C=CC(NCCC4=CN=CC=C4)=NN32; InChI=1S/C19H15N7/c20-12-14-3-5-16(6-4-14)19-24-23-18-8-7-17(25-26(18)19)22-11-9-15-2-1-10-21-13-15/h1-8,10,13H,9,11H2,(H,22,25)<br>

Current revision

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Contents

Open Source Malaria Series 4: The Triazolopyrazine (TP) Series

What's New

This series is currently active. (How to respond/input, for example if you want to suggest a molecule that should be made, is described in the Landing Page under "Join the Team")

Automatically updated list of to do items in this Series

Automatically updated list of compounds being made in this series

Most recent online meeting relevant to this series

Most recent PDF newsletter may be downloaded using this link.

Introduction

Preamble

The Triazolopyrazine Series is the newest of the OSM series. It was announced on September 10th 2013 and is sometimes referred to as the TP Series, or OSM Series 4.

Representative Series 4 Compound
Representative Series 4 Compound

The series arises from industrial work that cannot be fully disclosed which was followed by some hit-to-lead work funded directly by MMV and performed by a CRO which can.

A great deal of exploration of the series has been done, with significant diversity in the core and pendant groups. The series includes many potent compounds.

There is evidence from parasite ion regulation assays (below) that these compounds may be PfATP4 inhibitors. Such evidence distinguishes Series 4 from Series 1-3 where there was no experimental evidence for a mechanism of action.

As with everything involved in OSM, suggestions can be given in multiple ways.

Start of the Campaign

A briefing document written for MMV was the initial knowledgebase. This was accompanied by a PDF summary of pharmacokinetics and efficacy. This is being folded into the sections below, then supplemented.

The aim of the campaign at the outset was to improve the metabolic stability and the pharmacokinetic properties of this series in rat so as to meet the once-dosing criteria (TCP1) set by MMV. New chemistry directed towards blocking the putative metabolic sites was a major part of the research prior to the data being contributed to OSM.

Sources of Data on the TP Series

Initial briefing document (A minor error in the briefing document referring to the amides has been clarified.) Briefing document mostly folded into the below.
PDF summary of pharmacokinetics and efficacy. Needs folding into the below.
The current synthetic lab notebook
Raw Data Spreadsheet on Compounds Prior to Donation to OSM including available quantities of original samples
Summary of Data on Amides in the Series
Document from CRO describing synthetic chemistry of some members of series

Current Aims

1. Lead optimisation, to improve solubility and metabolic stability while maintaining potency.
2. Validation of PfATP4 activity as mechanism of action.

Current/Recent Activity

Last updated: Apr 4th 2014

1. Which compounds to make next? - see the most recent meeting for links to the discussions on these points. Compounds in the ether and amide sub-series are being made. Automatically-updated list of compounds being made now - some compounds may be from older series.

2. Does anyone possess compounds that could be relevant to this series already? It currently appears as though the answer is "no".

3. Resources needed: a) Chemists to make molecules, b) Experienced medicinal chemists to digest the data on this wiki and suggest new compounds to make. You can contact the OSM Consortium in many ways (go to the "Join the Team" page here.

Notable Points about the TP Series

  • Compounds in this series have been identified down to 16 nM potency.
  • Seems to have good in vitro HLM and hHEP stability Clint < 8.1 is compatible with 10 nM potency.
  • RLM remains stubbornly high, particularly for the more potent analogues translating to short half-lives in rat PK.
  • Series appears to have little polypharmacology or cytotoxicity.
  • Not so far challenged the hypothesis that rat metabolism may not be a great model for human metabolism for this series.
  • The series shows activity in Kieran Kirk’s PfATP4 assay which goes away for Pfal inactives in the series.

Concerns about the TP Series

  • Although dofetilide binding looks weak or nil, the series has shown activity in a patch clamp assay at Essen (1-10 uM) which is quite potent though with a window of >100 fold over Pfal potency.
  • In Kip Guy’s resistant mutants the picture is mixed, but there is still support for the idea that some members of the series are weaker in the resistant strains. The series has no or weak >>1uM activity against gametocytes, no activity against Winzeler’s Pb liver stage and may have weak activity against ookinetes but the dose-response data has not been completed.

Project Strands of Current Interest

The biggest issue is metabolic stability, as measured in rat in particular. There are few toxicity concerns. Thus possible future directions:

  • Small scale changes around the side chains, particularly phenethyl to attempt to balance potency and metabolism. Other possibilities: a) N is tolerated in the ring, hasn’t been explored much recently. b) Is 3,4-diF the best substitution pattern? c) Some evidence (eg MMV669848) that the phenethyl side chain can be rigidified, perhaps the iso-indoline of that compound could be improved on with other ring systems and by more optimal substitution of the aromatic benzene ring of the isoindoline. d) The amide MMV670944 is interesting and shows good RLM stability, but many other amides failed to match its potency.
  • Incorporation of a basic centre to increase volume as a potential fix for half-life. However, this might come at the expense of plasma concentration so would require high potency. Of the 29 compounds with a basic centre only one (MMV670437, below) has a measured potency < 100 nM (actually 44 nM).
MMV670437
MMV670437
  • More significant structural changes. Of the changes made to the basic skeleton, the most successful might be the recent evaluation of the substitution position changes (e.g., MMV670945), possibly in combination with modifying the disposition of the N atoms in the core. Related compounds have been made by others and it would be wise to incorporate the learnings from these series into any plans to explore this substitution pattern further. The first few compounds look similar in terms of metabolic stability.

SAR

Cycloaliphatic Triazole Substituent

Attempts at lowering the lipophilicity of the TP compounds by replacing the triazole aryl substituent with a cyclo(hetero)aliphatic group, linked either by the heteroatom or otherwise (e.g. piperidine, tetrahydropyran, indoline or isoindoline) lowered the potency against PfNF54, as did an aniline substituent.

Changes to the Triazole Substitution
Changes to the Triazole Substitution

Core Modification of Pyrazine Ring

Based on an assumption that the pyrazine moiety of the TP could undergo AO metabolism at positions alpha- to the nitrogen a few compounds were made with different R groups (Cl, Me, NH2, NEt2). However, all these compounds lost potency against PfNF54.

Changes to the Pyrazine Substitution
Changes to the Pyrazine Substitution

Core Modification of Triazole Ring

Two compounds based on imadazopyrazines were made (MMV669846 and MMV670250, below). Both showed reduced potency against PfNF54 vs. the corresponding TP compound. The RLM stability of one was found to be poor. Approx 20 structures were made with variations to the 6,5 core system. MMV669846 was the most potent. As most of the analogues were >1 μM potency, fewer were tested in RLM (quite a few in HLM). Of the 4 tested in RLM, the greatest stability had a Clint of 109, (HLM 9.5), several had HLM Clint 8 or less, particularly after moving or removing the N from the pyrazine ring.

Core Modification of Triazine Ring
Core Modification of Triazine Ring

(Note - original briefing document contained two entries for MMV669846 at this point with different potencies - need check of original spreadsheet, to verify the above is correct)

Transposing the Pyrazine side-chain

The side-chain on the pyrazine ring was shifted to the adjacent carbon. The chain length was varied (n= 0,1,2) and linked through either O or N.

Shifted Side Chain
Shifted Side Chain

Among the ethers, the phenethyl ether (X = O, n =2) showed good potency (Pfal IC50 33 nM) but a poor stability in RLM (Cl 100 mL/min/Kg)/

Shifted Ethers and Amines
Shifted Ethers and Amines

Pyrazine side-chain modifications - non amides

The phenethyl ether side-chain on the pyrazine ring was flagged as a metabolic hot-spot and so several strategies were adopted to mitigate this risk. The following findings were observed:

  • Changing the length of the side-chain to anything other than 3 atoms severely lowers potency against PfNF54
  • The linker atom to the pyrazine ring is crucial (O>>C>N)
  • Heteroatoms in the side-chain lowered potency with the exception of MMV669848 (featuring an isoindolino-methyl group on the pyrazine ring) although this compound had poor RLM stability.
  • Constraining the linear side-chain into ring systems (e.g, azetidines, pyrrolidines, pyrazoles) severely reduced potency.
MMV669848
MMV669848

A 2-naphthol substituent on the pyrazine ring showed reasonable potency against Pfal (IC50 114 nM) but suffered from poor RLM stability. Several hetero-analogs of 2-naphthol e.g. indole, indazole, quinoline, chroman, benzisoxazole, quinazoline, etc were made but all lost potency against the parasite.

The phenyethyl chain was replaced by an aromatic group in an attempt to mitigate the potential metabolism of the ethyl chain. Several compounds with a phenol substituent on the pyrazine ring were made. Some substituted phenolates were metabolically more stable in vitro as well as in vivo in rat, although with reduced potency against Pfal.

MMV669784
MMV669784

Considering that the benzylic position in the phenethyl side-chain is prone to metabolic oxidation, several compounds having mono- and di-substitution in the benzylic position were made. Di-Substitution lowered the potency considerably whereas mono-substitution with OMe, OCHF2, CH2OH, NMe2 groups retained good potency. Additional substitution alpha- to the ether oxygen led to complete loss of potency. The alpha-OCHF2 compound MMV670652, with a p-CN-phenyl group on the triazole ring, showed better RLM stability (cLogP effect).

Pyrazine side-chain modifications - amides

A small library of amides (including the m-Cl benzylamide, MMV668958) showed promising potency. However, other amides (derived from aliphatic or anilines) either showed lower potency or were inactive. The RLM of MMV668958 was poor - perhaps due to benzylic oxidation. Alpha-substitution at the benzylic position or constraining the benzylamine into an aminoindane did not improve potency. Several attempts to make aniline-amides with improved potency against Pfal failed. Loading the aniline ring with lipophilic substituents marginally improved potency but led to poor RLM stability.

Amides Synthesised and Tested pre-OSM
Amides Synthesised and Tested pre-OSM

The p-Cl benzanilide MMV670246, although not active against PfNF54, showed good RLM stability perhaps due to lack of benzylic metabolism. However, its rat PK showed high clearance. The potent m-Cl (MMV669542) analog had poor RLM stability.

Further PK Details on MMV670246
Further PK Details on MMV670246

Pharmacokinetics

Only one compound in this Series first made at the CRO was measured in rat PK and that was the relatively weak amide MMV670246. The curve is shown for oral & IV legs & parameters are below.

MMV670246 in rat PK
MMV670246 in rat PK

[Data for MMV670246 in rat PK]

One of the compounds with better in vitro balance is MMV670652 with potency at 17 nM, HLM Clint < 8 ul/min/mg and RLM Clint at 30 ul/min/mg. This compound has not been in rat PK. Additionally it may be possible to improve potency by synthesis of the more potent enantiomer.

MMV670652
MMV670652

A PDF summary of pharmacokinetics and efficacy provided as an initial source contained efficacy information for the compound MMV639565.

MMV639565
MMV639565

The therapeutic efficacy of MMV639565 against P falciparum growing in peripheral blood of NODscidIL2Rγnull mice engrafted with human erythrocytes (po dosing, qd for 4 days) is shown below. A rapid parasite clearance - ED90 6.3 mg/kg was recorded.

MMV639565 Therapeutic Efficacy
MMV639565 Therapeutic Efficacy

Values in brackets indicate dose corrected according to quality control of formulation.

Other Observations

As would be expected HLM vs. RLM shows a general correlation with approx 4-fold shift on average. However, for most of the more potent analogs, this increases to over 10-fold. The figure below shows the 4 sub 30nM compounds with HLM & RLM measured.

HLM vs RLM
HLM vs RLM

Lipophilicity & lipophilic efficiency:

Few compounds achieve a lipophilic efficiency (as measured by pIC50 – AlogP) of greater than 4.0

Lipophilic efficiency
Lipophilic efficiency

Potential next steps - suggested in original pre-OSM briefing document

The series has good potency and in vivo efficacy with few toxicity concerns. The biggest issue is metabolic stability, as measured in rat in particular. Some possible future directions include:

  • Small scale changes around the side chains, particularly phenethyl to attempt to balance potency and metabolism
  • N is tolerated in the ring, hasn’t been explored much recently
  • Is 3,4-diF the best substitution pattern ?
  • Some evidence (eg. MMV669848) that the phenethyl side chain can be rigidified, perhaps the iso-indoline of that compound could be improved on with other ring systems and by more optimal substitution of the aromatic benzene ring of the isoindoline.
  • The amide MMV670944 is interesting and shows good RLM stability, but many other amides failed to match its potency
  • Incorporation of a basic centre to increase volume as a potential fix for half-life. However, this might come at the expense of plasma concentration so would require high potency. In addition of the 29 compounds with a basic centre only one has a measured potency < 100nM.
MMV670437
MMV670437
  • More significant structural changes. Of the changes made to the basic skeleton, the most successful might be the recent evaluation of the substitution position changes (eg MMV670945), possibly in combination with modifying the disposition of the N atoms in the core. Related compounds have been made by others and it would be wise to incorporate the learnings from these series into any plans to explore this substitution pattern further. The first few compounds look similar in terms of metabolic stability.

Possible PfATP4 Activity Deduced from Parasite Ion Regulation Assays

The following five compounds were evaluated in parasite ion regulation assays in the Kirk Laboratory; the hypothesis is that PfATP4 is a Na+ ATPase that exports Na+ and imports H+ (or equivalent) and that the effects of the compounds on Na+ concentration and pH are attributable to inhibition of this activity. Structures, potency, metabolism/solubility and raw PfATP4 assay data are here.

The Five Compounds Initially Evaluated for Possible PfATP4 Activity
The Five Compounds Initially Evaluated for Possible PfATP4 Activity

MMV669000: no (potency: inactive)

MMV669304: yes (potency: 280 nM)

MMV669360: yes (potency: 356 nM)

MMV669542: yes (potency: 185 nM)

MMV669848: yes (potency: 114 nM)

MMV669000 did not dissipate the plasma membrane Na+ gradient or increase the plasma membrane pH gradient consistent with it not inhibiting PfATP4 at the concentration tested.

The other compounds dissipated the plasma membrane Na+ gradient and increased the plasma membrane pH gradient at a concentration of 2 μM, consistent with them being PfATP4 inhibitors.

(i.e. note the correlation: compound inactive in these assays is the inactive analog in the plasmodium screen)

Physicochemical/metabolism

The following compounds were sent (associated discussion) and data were obtained.

Compounds Evaluated for Solubility and Metabolic Clearance
Compounds Evaluated for Solubility and Metabolic Clearance

Solubility is low and improvement should be one of the goals for lead optimisation. The metabolic stability is not a mouse specific event and needs to be improved to deliver a candidate. Met ID data may help in the design of new analogs with improved stability, but we could also look at correlations with Log D etc. to drive new target design)

Metabolism ID

The following compounds were sent and data were obtained.

Compounds Evaluated for Metabolic ID
Compounds Evaluated for Metabolic ID

The data suggest that the core heterocyclic ring is subjected to an oxidation (though it is not clear the extent to which the compound is oxidized, only that certain peaks are present). The nature of the side chain appeared to have little influence on the results, suggesting alteration of the side chain is probably not the way to stop turnover, meaning it is likely that amides made in this series in the near future will also suffer from this problem. Nevertheless the efficacy was seen as sufficiently promising that amides remain attractive as they are. i.e. they are sufficiently robust for this point in the project. Residual question: what is the mechanism of the clearance (oxidation)? Is it CYP mediated? Open action item here.

hERG Data

MMV669844 and MMV670944 (OSM-S-175) were evaluated in a hERG assay, with the data indicating issues with both compounds that will need to be addressed.

Synthetic Chemistry

Synthetic Design

The approach to the synthesis of Series 4 up until early 2014 has involved the functionalisation of an existing pyrazine core, followed by a cylization reaction to give the triazolopyrazine.

Literature examples of cyclization to give triazolopyrazine (Tom to insert one example).

Synthetic chemistry for OSM in Jan/Feb 2014 has been focussed on two main targets in Series 4 (the triazolopyrazines) - the ether linked compounds and the amide compounds, shown below along with specific examples of known, potent compounds in each series.

Examples of ether and amide target molecules.
Examples of ether and amide target molecules.

Synthesis of the Ether-Linked Series

The ether series contains many active compounds. MMV670652 shown above was evaluated as a racemate, and Alice has been attempting a resynthesis with a view to the separation of enantiomers to evaluate the properties of the active enantiomer. Here's the original issue for the synthesis of compound MMV670652, as well as the first discussion of how we might separate the enantiomers, and a newer issue created after a recent meeting. One of the complexities of the synthesis is the difluoromethylation step (original summary of problem is here). It's not clear that this group is needed - there are related structures that could be tried, and compounds lacking this group (and indeed the whole stereogenic centre) are still active.

A key synthetic stumbling block has been the union of the two halves of the molecule - an alcohol (2) and the heterocycle chloride (7), for example in this recent reaction. The reactions were for a long time not proceeding cleanly, and this was also found by the CRO that first did it - report was posted here. Jo Ubels performed a lit survey and found conditions involving a crown ether that now appear to provide the desired ether. The synthesis of the chiral alcohol fragment needed specifically for MMV670652, via 1, requires a source of cyanide, and we're currently out of stock in Sydney. Thus currently simpler alcohols (i.e. ArCH2CH2OH) are being used in this step. A new enantioselective approach is also being developed by Alice in the meantime.

Synthetic route to ether.
Synthetic route to ether.

Another synthetic challenge being addressed is the efficiency of the cyclization to give 7 from the precursor 6. This has been achieved, but not in the ca. 50% yields reported by the original CRO. A new Honours student in the group, Jo Ubels, is now looking at this step to work out what is going on and how we can make this reaction more efficient and this has been the subject of research by an Edinburgh student working with Patrick Thomson, Devon Scott. There appear to be significant issues to do with solubility of the material throughout the reaction. The synthesis of the precursor 7 is straightforward.

Synthesis of the Amide-Linked Series

The amide series is interesting because there are potent compounds with such a structure that appear to have desirable PK properties, but potency needs to be enhanced. Following an analysis of commercially-available primary and secondary amines that could be incorporated, the likely physical properties of the resulting compounds as well as an analysis of those amines we had to hand in the building, synthesis was started (scheme below). It has been noted that an efficient approach here would be to use the same intermediate as for the ether series (7, above) and invent a new carbonylation method (original issue, Inga's lit survey, Alice's original appeal for a collaborator and a summary of the associated discussion.

For the moment, however, it was decided to pursue an alternative route wherein the extra carbon atom is incorporated from the start using acid 9. This molecule is commercially available but is expensive, but we now have routes to it in the lab. Patrick and Inga planned the synthesis from the pyrazine acid and then Inga completed a synthesis of it, as has Eduvie using the same approach (though data are missing for these latter experiments). Sabin has successfully demonstrated his very nice alternative route to this compound in one step from the methyl precursor (data). Tom is now scaling up the Patrick/Eduvie/Inga approach to bring through more material.

For the subsequent coupling of the pyrazine acid with an amine, Tom and Alice have identified a clean coupling reaction to give the amide 10 using T3P that worked much better than an attempt to go via the acid chloride, and Alice has seen excellent preliminary results in the coupling of a related amide with hydrazine, condensation with aldehydes (this chemistry has been surveyed by Inga) and is now attempting the crucial cyclization reaction to give what would be the final amide. Should this work, we would then have access to a wide range of new members in the amide series. Here is one of Tom's current "Being Synthesised Now" Github posts in this series. Here is one of Alice's where she is altering the northeast part of the molecule. There was a suggestion that hydrazone isomers may be present and may be differently reactive towards the oxidative cyclisation conditions (GH issue 97).

Synthetic route to amide.
Synthetic route to amide.

Chirality/Stereogenic Centres in This Series

MMV670652 shown above was evaluated as a racemate, and there was interest in separating enantiomers: original issue then discussion of how we might separate the enantiomers, and a more recent issue. It is not clear that this group is needed - there are related structures that could be tried, and compounds lacking this group (and indeed the whole stereogenic centre) are still active. MMV669844 (CH3 rather than CHF2) was synthesised using a Sharpless epoxidation on the precursor styrene, but there are no data on the enantiomeric excess of the product - it is shown as a single enantiomer in the original briefing document.

Other Sources of Compounds Relevant to this Series

(Original community request)
The compound series appears to be novel/unusual with little similarity to compounds in PubChem/Surechem, though Chris Southan (in the previous link) found a related compound (CID 7118230) with reported antimalarial activity, with a structure similar to those described above for modification of the triazolopyrazine core. Jo Ubels also carried out a search and found few compounds similar to Series 4.
A similarity was noted between Series 4 compounds and a compound from Novartis likely active against vivax. Paul Willis suggested the mechanism of action of the Novartis compounds was likely PI4K - data and reasoning given in this lab book entry. Chris Southan modelled the OSM and Novartis compounds, which provides additional evidence for a different mechanism of action.

Strings for Google

Use this section to paste strings to make the page more discoverable.


MMV668955 FC(F)OC(C=C1)=CC=C1C2=NN=C3N2C(CCO)CNC3; InChI=1S/C14H16F2N4O2/c15-14(16)22-11-3-1-9(2-4-11)13-19-18-12-8-17-7-10(5-6-21)20(12)13/h1-4,10,14,17,21H,5-8H2
MMV668958 FC(F)OC(C=C1)=CC=C1C2=NN=C3N2C(C(NCC4=CC(Cl)=CC=C4)=O)=CN=C3; InChI=1S/C20H14ClF2N5O2/c21-14-3-1-2-12(8-14)9-25-19(29)16-10-24-11-17-26-27-18(28(16)17)13-4-6-15(7-5-13)30-20(22)23/h1-8,10-11,20H,9H2,(H,25,29)
MMV668960 FC(F)OC(C=C1)=CC=C1C2=NN=C3N2CCNC3; InChI=1S/C12H12F2N4O/c13-12(14)19-9-3-1-8(2-4-9)11-17-16-10-7-15-5-6-18(10)11/h1-4,12,15H,5-7H2
MMV668962 FC(F)OC(C=C1)=CC=C1C2=NN=C3N2CCN(C(CN)=O)C3; InChI=1S/C14H15F2N5O2/c15-14(16)23-10-3-1-9(2-4-10)13-19-18-11-8-20(12(22)7-17)5-6-21(11)13/h1-4,14H,5-8,17H2
MMV669000 O=C(N1CC(C=CC=C2)=C2C1)C3=CN=CC4=NN=C(C5=CC=C(OC(F)F)C=C5)N43; InChI=1S/C21H15F2N5O2/c22-21(23)30-16-7-5-13(6-8-16)19-26-25-18-10-24-9-17(28(18)19)20(29)27-11-14-3-1-2-4-15(14)12-27/h1-10,21H,11-12H2
MMV669025 FC1=C(F)C=CC(CCOC2=CNC(C3=NN=C(C4=CC=C(OC(F)F)C=C4)N32)=O)=C1; InChI=1S/C20H14F4N4O3/c21-14-6-1-11(9-15(14)22)7-8-30-16-10-25-19(29)18-27-26-17(28(16)18)12-2-4-13(5-3-12)31-20(23)24/h1-6,9-10,20H,7-8H2,(H,25,29)
MMV669304 FC(F)OC(C=C1)=CC=C1C2=NN=C3C=NC=C(CCCC4=CC=CC=C4)N32; InChI=1S/C21H18F2N4O/c22-21(23)28-18-11-9-16(10-12-18)20-26-25-19-14-24-13-17(27(19)20)8-4-7-15-5-2-1-3-6-15/h1-3,5-6,9-14,21H,4,7-8H2
MMV669310 FC(F)OC(C=C1)=CC=C1C2=NN=C3N2C(CCCC4=CC=CC=C4)CNC3; InChI=1S/C21H22F2N4O/c22-21(23)28-18-11-9-16(10-12-18)20-26-25-19-14-24-13-17(27(19)20)8-4-7-15-5-2-1-3-6-15/h1-3,5-6,9-12,17,21,24H,4,7-8,13-14H2
MMV669360 FC(F)OC(C=C1)=CC=C1C2=NN=C3C=NC=C(COCC4=CC=C(F)C(F)=C4)N32; InChI=1S/C20H14F4N4O2/c21-16-6-1-12(7-17(16)22)10-29-11-14-8-25-9-18-26-27-19(28(14)18)13-2-4-15(5-3-13)30-20(23)24/h1-9,20H,10-11H2
MMV669542 FC(F)OC(C=C1)=CC=C1C2=NN=C3C=NC=C(C(NC4=CC=CC(Cl)=C4)=O)N32; InChI=1S/C19H12ClF2N5O2/c20-12-2-1-3-13(8-12)24-18(28)15-9-23-10-16-25-26-17(27(15)16)11-4-6-14(7-5-11)29-19(21)22/h1-10,19H,(H,24,28)

MMV669844 [C@H](COc1cncc2n1c(nn2)c1ccc(cc1)C#N)(c1cc(c(cc1)F)F)OC

MMV669846 FC1=C(F)C=CC(CCOC2=CN=CC3=NC=C(C4=CC=C(Cl)C=C4)N32)=C1; InChI=1S/C20H14ClF2N3O/c21-15-4-2-14(3-5-15)18-10-25-19-11-24-12-20(26(18)19)27-8-7-13-1-6-16(22)17(23)9-13/h1-6,9-12H,7-8H2
MMV669848 FC(F)OC(C=C1)=CC=C1C2=NN=C3C=NC=C(CN4CC(C=CC=C5)=C5C4)N32; InChI=1S/C21H17F2N5O/c22-21(23)29-18-7-5-14(6-8-18)20-26-25-19-10-24-9-17(28(19)20)13-27-11-15-3-1-2-4-16(15)12-27/h1-10,21H,11-13H2
MMV670246 FC(F)OC(C=C1)=CC=C1C2=NN=C3N2C(C(NC4=CC=C(Cl)C=C4)=O)=CN=C3; InChI=1S/C19H12ClF2N5O2/c20-12-3-5-13(6-4-12)24-18(28)15-9-23-10-16-25-26-17(27(15)16)11-1-7-14(8-2-11)29-19(21)22/h1-10,19H,(H,24,28)
MMV670250 FC1=C(F)C=CC(CCOC2=CN=CC3=CN=C(C4=CC=C(Cl)C=C4)N32)=C1; InChI=1S/C20H14ClF2N3O/c21-15-4-2-14(3-5-15)20-25-11-16-10-24-12-19(26(16)20)27-8-7-13-1-6-17(22)18(23)9-13/h1-6,9-12H,7-8H2
MMV670437 FC(F)OC(C=C1)=CC=C1C2=NN=C3C=NC=C(OC[C@H](N(C)C)C4=CC(F)=C(F)C=C4)N32; InChI=1S/C22H19F4N5O2/c1-30(2)18(14-5-8-16(23)17(24)9-14)12-32-20-11-27-10-19-28-29-21(31(19)20)13-3-6-15(7-4-13)33-22(25)26/h3-11,18,22H,12H2,1-2H3/t18-/m0/s1

MMV670936 C(COc1cncc2n1c(nn2)c1cnc(cc1)C(F)(F)F)c1cc(c(cc1)F)F

MMV670944 (OSM-S-175) FC(F)OC(C=C1)=CC=C1C2=NN=C3N2C(C(NC4=CC=NC(F)=C4)=O)=CN=C3; InChI=1S/C18H11F3N6O2/c19-14-7-11(5-6-23-14)24-17(28)13-8-22-9-15-25-26-16(27(13)15)10-1-3-12(4-2-10)29-18(20)21/h1-9,18H,(H,23,24,28)
MMV670945 FC(F)OC(C=C1)=CC=C1C2=NN=C3N2C=C(OCCC4=CC=C(F)C(F)=C4)N=C3; InChI=1S/C20H14F4N4O2/c21-15-6-1-12(9-16(15)22)7-8-29-18-11-28-17(10-25-18)26-27-19(28)13-2-4-14(5-3-13)30-20(23)24/h1-6,9-11,20H,7-8H2
MMV670946 FC(F)OC(C=C1)=CC=C1C2=NN=C3N2C=C(OCC4=CC=C(F)C(F)=C4)N=C3; InChI=1S/C19H12F4N4O2/c20-14-6-1-11(7-15(14)21)10-28-17-9-27-16(8-24-17)25-26-18(27)12-2-4-13(5-3-12)29-19(22)23/h1-9,19H,10H2
MMV670949 FC(F)OC(C=C1)=CC=C1C2=NN=C3N2C=C(NCC4=CC=C(F)C(F)=C4)N=C3; InChI=1S/C19H13F4N5O/c20-14-6-1-11(7-15(14)21)8-24-16-10-28-17(9-25-16)26-27-18(28)12-2-4-13(5-3-12)29-19(22)23/h1-7,9-10,19,24H,8H2
MMV671655 FC(F)OC(C=C1)=CC=C1C2=NN=C3N2C=C(NC4=CC=C(F)C(F)=C4)N=C3; InChI=1S/C18H11F4N5O/c19-13-6-3-11(7-14(13)20)24-15-9-27-16(8-23-15)25-26-17(27)10-1-4-12(5-2-10)28-18(21)22/h1-9,18,24H
MMV671926 FC(F)OC(C=C1)=CC=C1C2=NN=C3N2C=C(OC4=CC=C(F)C(F)=C4)N=C3; InChI=1S/C18H10F4N4O2/c19-13-6-5-12(7-14(13)20)27-16-9-26-15(8-23-16)24-25-17(26)10-1-3-11(4-2-10)28-18(21)22/h1-9,18H
MMV671927 FC(F)OC(C=C1)=CC=C1C2=NN=C3N2C=C(NCCC4=CC=C(F)C(F)=C4)N=C3; InChI=1S/C20H15F4N5O/c21-15-6-1-12(9-16(15)22)7-8-25-17-11-29-18(10-26-17)27-28-19(29)13-2-4-14(5-3-13)30-20(23)24/h1-6,9-11,20,25H,7-8H2
MMV672619 FC(F)OC(C=C1)=CC=C1C2=NN=C3N2C=C(O[C@@H]4C(N(C5CC5)[C@@H]4C6=CC(F)=C(F)C=C6)=O)N=C3; InChI=1S/C24H17F4N5O3/c25-16-8-3-13(9-17(16)26)20-21(23(34)33(20)14-4-5-14)36-19-11-32-18(10-29-19)30-31-22(32)12-1-6-15(7-2-12)35-24(27)28/h1-3,6-11,14,20-21,24H,4-5H2/t20-,21+/m1/s1
MMV672939 FC(F)OC(C=C1)=CC=C1C2=NN=C3C=CC(OCCC4=CC(F)=C(F)C=C4)=NN32; InChI=1S/C20H14F4N4O2/c21-15-6-1-12(11-16(15)22)9-10-29-18-8-7-17-25-26-19(28(17)27-18)13-2-4-14(5-3-13)30-20(23)24/h1-8,11,20H,9-10H2
MMV672942 FC(F)OC(C=C1)=CC=C1C2=NN=C3C=CC(NCCC4=CC=C(F)C=C4)=NN32; InChI=1S/C20H16F3N5O/c21-15-5-1-13(2-6-15)11-12-24-17-9-10-18-25-26-19(28(18)27-17)14-3-7-16(8-4-14)29-20(22)23/h1-10,20H,11-12H2,(H,24,27)
MMV672990 FC(F)OC(C=C1)=CC=C1C2=NN=C3N2C=C(NC(CC4=CC=CC(Cl)=C4)=O)N=C3; InChI=1S/C20H14ClF2N5O2/c21-14-3-1-2-12(8-14)9-18(29)25-16-11-28-17(10-24-16)26-27-19(28)13-4-6-15(7-5-13)30-20(22)23/h1-8,10-11,20H,9H2,(H,25,29)
MMV672992 N#CC(C=C1)=CC=C1C2=NN=C3C=CC(NCCC4=CN=CC=C4)=NN32; InChI=1S/C19H15N7/c20-12-14-3-5-16(6-4-14)19-24-23-18-8-7-17(25-26(18)19)22-11-9-15-2-1-10-21-13-15/h1-8,10,13H,9,11H2,(H,22,25)

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