What is KCGSv2.0?

The Kinase Chemogenomic Set (KCGSv2.0) is a collection of 295 potent and selective small molecule ATP-competitive inhibitors that were produced in drug discovery programs targeting protein kinases. Each inhibitor has been profiled for binding against >200 human kinases at DiscoverX. Each inhibitor has a K_d < 100 nM for its target kinase with a selectivity index S₁₀ < 0.04 at 1 µM. In total, KCGSv2.0 indexes 262 human kinases including many poorly studied (dark) kinases. KCGSv2.0 is available from the SGC in aliquots of 1 µL of a 10 mM solution in DMSO.

How do I find more information about KCGSv2.0?

An open access scientific paper and supporting information describing KCGSv2.0 with details of the kinases covered and its use in cell-based assays has been published in the International Journal of Molecular Sciences.

Where can I get the chemical structures and kinase inhibition data?

Details on how to access the data are provided here KCGS data page. The chemical structures of the inhibitors and the kinase activity data are available on our website Random Acts of Kinase.

How do I obtain KCGSv2.0?

Detailed instructions for requesting the set are posted on our web page. UNC Researchers have access to KCGSv2.0 through Core Serices. All other researchers have access to KCGSv2.0 through CancerTools.org.

Why are you charging for KCGSv2.0?

KCGSv2.0 is currently a finite resource that is available in limited supply through donation of kinase inhibitors from pharma companies and academic labs. We will use all funds raised through distribution of KCGSv2.0 to finance resynthesis of the kinase inhibitors so we can continue to make it available as a public resource.

How did you decide the $3100 processing and distribution fee?

A survey of recipients of PKIS and PKIS2 indicated that the majority would have paid between $1000-$5000 for access to the sets. All funds raised through distribution of KCGSv2.0 will be used to support resynthesis of the kinase inhibitors so we can provide the set for years to come.

What if I cannot afford the processing and distribution fee?

We will gladly work with you to submit grant applications to raise the funds. It is likely that the KCGSv2.0 processing and distribution fee will be only a fraction of the operational cost of your proposed experiment.

How should we store KCGSv2.0 upon receipt?

We recommend storing the plate between -20 and -80°C until use. Researchers are advised to avoid multiple freeze-thaw cycles with the KCGSv2.0 plate to ensure best results.

What concentration range should be used when running a cellular screen using the KCGSv2.0 library?

KCGSv2.0 contains potent kinase inhibitors that have been profiled for selectivity at 1 μM. We recommend cellular screens be conducted no higher than 1 μM. If assay throughput can accommodate it, another option is to conduct the initial screening at 2-3 different concentrations; for example 10 nM, 100 nM, and 1 μM.

How are KCGSv2.0 compounds solubilized?

All KCGSv2.0 compounds are dissolved in DMSO and dispensed as 1 μL of a 10 mM stock solution.

What kind of plate is KCGSv2.0 dispensed into?

KCGSv2.0 is dispensed in a Greiner #781280 V-bottom polypropylene 384-well plate as 1 μL of a 10 mM stock solution.

What is the total capacity of the wells in the KCGSv2.0 plate?

The capacity of the KCGSv2.0 wells is 130 μL. It is recommended to fill up to 100 μL if automation is involved.

Is there a recommended dilution protocol for the KCGSv2.0 plate?

When ready to dilute the plates, we recommend bringing the KCGSv2.0 plate to room temperature, spinning down all liquid, diluting and agitating. We recommend diluting with DMSO. For example, adding 9 μL of DMSO to the 1 μL of 10 mM stock will yield 10 μL of 1 mM stock in your KCGSv2.0 plate. A 1000-fold dilution of the stock plate in assay media will yield 10 mL of a 1 μM solution containing 0.1% DMSO.

How much can I request?

We dispense KCGSv2.0 as 1 µL of a 10 mM solution in DMSO. As KCGSv2.0 is currently a limited resource and you require more than the standard dispensement we will work with you to define the minimal volume necessary to enable the proposed experiments and decide if we can meet that need.

Is there space for controls on the KCGSv2.0 plate?

The last two columns of every plate are left intentionally blank for control wells when plating KCGSv2.0.

Which compounds are contained in my plate?

Physical and electronic plate maps are included with every shipment. We will be providing plate maps on our website that people can access using the plate number they receive as well.

What is known about the stability of compounds in KCGSv2.0?

The compounds in KCGSv2.0 are not known to have any light or air sensitivities. A QC plate was analyzed prior to dispensing the KCGSv2.0 plates. However, researchers are advised to avoid multiple freeze-thaw cycles with the KCGSv2.0 plate to ensure best results.

Does SGC-UNC have general toxicity data for the KCGSv2.0 library?

Details of the general toxicity at 10 µM inhibitor concentration in HeLa cells can be found in the open access paper https://doi.org/10.3390/ijms22020566 The effect of the inhibitors at 1.0 µM concentration on growth rate across 17 cell lines is also detailed in the manuscript.

What is the kinome coverage of KCGSv2.0?

Using data collected across 401 human kinases, KCGSv2.0 covers 54% of the screenable kinome. Each inhibitor has a Kd <100 data-preserve-html-node="true" nM for its target kinase with a selectivity index S10 <0.04 data-preserve-html-node="true" at 1 µM. There are no highly promiscuous kinase inhibitor in KCGS.

How do I share data generated with KCGSv2.0?

The data can be published in scientific literature, shared by emailing it to us, or by connecting with a data repository such as ChEMBL.

Do I need to include SGC-UNC scientists on any publications that arise from use of the set?

We do not feel that simple supply of compounds justifies inclusion as coauthors. If we add something more to a collaborative effort, please use normal academic standards for coauthorship. However, all publications should identify the source of KCGSv2.0 as the SGC-UNC and you should send a copy of the citation to us upon publication. We will post a list of all publications arising from the use of KCGSv2.0 on our website.

What do I need to put in the Acknowledgments section of a publication?

We recommend using the following text “KCGSv2.0 was supplied by the SGC-UNC and contains kinase inhibitors that were made available by GlaxoSmithKline, Takeda, Pfizer, Boehringer Ingelheim, Vertex, Harvard University, Cancer Research UK, and the SGC.”

What are the requirements for using the set?

Recipients of KCGSv2.0 must use the set within 90 days. Recipients of KCGSv2.0 will be identified on our website. You must communicate results to us and make all data derived from use of the sets publically available within a reasonable amount of time.

I have some interesting results. Can I get additional quantities of compounds of interest?

We try to supply additional quantities compounds when possible from our internal supplies, but these are often limited. However, we will always provide options for obtaining additional quantities of the compounds through resynthesis.

I have some interesting results. Can I get analogs of compounds of interest?

We have analogs of many of the kinase inhibitors in KCGS that may be useful for establishing structure-activity relationships around a kinase of interest.

Where can I get the chemical structures and kinase inhibition data?

Details on how to access the data are provided here

I want to pursue an in vivo experiment with a molecule from KCGSv2.0. Is this possible?

KCGSv2.0 is formatted for screening in cells and little is known about the pharmacokinetic properties of the individual inhibitors. If you plan to use a KCGS compound in animals, evidence of an appropriate institutional animal care and use committee (IACUC or equivalent) protocol approval is required before we can consider a request of this nature. Quantities of individual compounds maybe severely limited and ultimately preclude supply for an in vivo study.

Why does the plate map show a space in the H7 position, but the well isn't empty?

The compound in there should be ignored. It was added in error, so we left this spot as a blank on the plate map.

How do I obtain the (up to) 5 x 1 µL of 10 mM DMSO cherry picks?

Please send the details of your request and shipping address to SGC-UNC@unc.edu. These are free to purchasers of the KCGSv2.0 set.