A variety of GPCR functional assay methods are available, such as Ca2+ assay, cAMP assay, β-arrestin assay, and reporter gene assay. However, some are clumsy, costly, and time-consuming, and others work with only some GPCRs. There has been no silver bullet … until now. Our proprietary assay platform has overcome limitations of existing methods and made all GPCRs accessible for drug discovery.

About GPCR

GPCRs as the Biggest Drug Target

Nat Rev Drug Discov 2017;16:829-842

G-protein-coupled receptors (GPCRs) comprise the largest class of membrane receptors and amount to approximately 800 in the human genome (left pie chart). Many of them are crucial in biology and medicine. ~200 of GPCRs respond to endogenous mediators and regulate a wide range of physiological parameters from blood pressure and blood glucose levels to cognitive functions. ~500 are sensory receptors sensing smells, tastes, and other modalities of stimulation. The remaining 100 are orphan, i.e., their ligands are yet to be identified.

Drug Browser. GPCRdb.

Although receptors for endogenous mediators are the most promising group of GPCRs in drug development, only a fraction of them have drugs to target them (right pie chart). Furthermore, recent studies have shown quite a few orphan receptors and sensory receptors to play vital roles in a variety of pathophysiology. Altogether, there remain enormous untapped opportunities with the vast majority of GPCRs.

Drugs have been developed for only a small part of GPCRs. The chart above ranks GPCRs in the ascending order according to the number of targeting drugs (the more drugs a GPCR has, the lower it sits). For example, DRD2 at the bottom has more than 80 approved drugs and compounds under R&D, but as you go up, the number of drugs per receptor drops quickly. The top 100 GPCRs keep almost all GPCR drugs to themselves, and the remaining hundreds are left out with no drugs.

GPCR ranking: Number of receptor-targeted drugs

All GPCRs are not made equal. As illustrated in "About GPCR," some GPCRs have dozens of drugs targeting them w […]

Barriers to GPCR Drug Discovery

One of the barriers to GPCR drug discovery is lack of satisfactory assays.

  • Binding assays are unreliable with many false-positives and false-negatives when it comes to GPCRs
  • No functional assay is adequate.
    • Only a small number of GPCRs are readily amenable to reporter gene assays, and G12/13- and Gi/o-coupled receptors are notoriously intractable.
    • Other functional assays cost a lot of time, work, and/or money. e.g., Western blotting

Core Technology

To overcome the barriers of GPCR drug discovery and promote drug development, we have innovated a high-performance functional assay.

Key Features

  • Remarkable sensitivity and S/B ratio
  • An exhaustive library of 313 human GPCRs
  • Works for all receptors coupled with any major Gα proteins incliding Gq, Gs, Gi/o, and G12/13
  • Makes orphan receptors practical targets.
  • Screens for agonists, antagonists, reverse agonists, PAM, and NAM

Components

The core technology for our cell-based GPCR functional assay consists of sensitive and versatile test cells, a Gα protein cocktail, and an exhaustive human GPCR library.

Proprietary Assay Cells

Outstandingly sensitive, specific, and versatile

  • Equipped with an artificial gene circuit to specifically amplify GPCR-induced intracellular signals
  • Works for all major Gα proteins including Gq, Gs, Gi/o, and G12/13
  • Verified with 99% (208 out of 210) of non-orphan GPCRs
  • Also works with other membrane receptors such as receptor tyrosine kinases
  • International patent filed and exclusively licensed from the University of Tokyo to Tanso Biosciences

Gα protein cocktail

This allround Gα protein cocktail complements our test cells and enables functional assay for any GPCR under a single protocol even without knowledege of the Gα protein the target receptor is coupled with.

Comprehensive human GPCR library

Industry-leading collection of 313 human GPCRs including 218 non-sensory receptors and 95 orphan receptors. Use of native sequences minimizes signal artifacts.

GPCR Catalog

We have all the 313 human non-sensory GPCRs in native sequences and provide functional assay for them in a sin […]

Validation

Our assay has yielded consistent readouts with results from existing methods and at high sensitivity and S/B ratio.

AGTR1, a peptide-ligand receptor

Tanso's method

Inositol phosphate formation

J Biol Chem. 1995;270:12846-50

Binding assay

LigandKi (nM) ± s.e.m.n
Sar1-angiotensin II0.38 ± 0.073
Sar1, Ala8-angiotensin II0.43 ± 0.083
Sar1, Ile8-angiotensin II0.46 ± 0.033
Ile7-angiotensin III1.1 ± 0.33
Angiotensin II1.6 ± 0.23
Angiotensin III5.4 ± 0.53
Dup7536.3 ± 0.93
Angiotensin I74 ± 123
Arg8-vasopressin>1,0002
Endothelin>1,0002
PD123177>10,0002

Nature. 1991;351:233-6

GPR91, a low molecular ligand receptor

In our assay, GPR91 responded to its agonist at high sensitivity and S/B ratio and consistently with the results from existing methods.

Tanso's method

Flt3, a receptor tyrosine kinase (RTK)

Tanso's method

Cells from the same line of test cells were used in this assay as in assays for GPCRs, and Flt3 was transiently transfected into cells in the same way as GPCRs.

Cell proliferation

Recombinant Human Flt-3 Ligand Protein. R&D System

FAQ

Do your test cells amplify noise as well as signals?

The artificial gene circuit in our cells is designed to amplify signals quadratically while boosting noise only by ~two-fold. The cells have responded to stimuli with >100-fold signal/background ratios and small standard errors in many GPCRs(see Validation).

Can your test cells examine biased agonism?

No, you need in vivo experiments, not cell-based assay, to test if compounds act in a biased way since biased agonism is defined by the expression pattern of Gα proteins in the target tissue.

Are your test cells capable of expressing orphan GPCRs in the plasma membrane when they are transfected?

It is essential to make sure that introduced orphan GPCRs are present in the plasma membrane all the more because they do not have agonists to prove that pharmacologically. We are testing each orphan receptor now.

Do you have animal GPCR panels?

We are expanding our library to mouse and rat GPCRs. If you are interested in a particular receptor, we can prioritize cloning it and will be ready to assay it in a few weeks.

Contact Us

Send us a message via the form below or write to us at info@tansobio.com.

Service Flow Chart

Request for Quotation

Tell us about the types of tests you need, the number of compounds, the receptors of interest, and other preferences. A non-disclosure agreement is optional.

STEP
1

Order

A non-disclosure agreement is optional.

STEP
2

Compound Shipment

The client ships test compounds to our laboratory. We usually perform assays and write a report in three weeks from the receipt of the compounds.

STEP
3

Receipt of the Report

We provide the report and raw data for the client via email or a cloud service.

STEP
4

Inquiry & RFQ Form