VDP-132 [Funding] Mario Cordero - Multi-inflammasome inhibitors to treat aging and age-related diseases

[VDP-132] Mario Cordero - Multi-inflammasome inhibitors to treat aging and age-related diseases

One-liner: Development of multi-inflammasome inhibitors as a potential treatment for age-related diseases.

Longevity Dealflow WG team

  • Senior Reviewers: Review completed. 5 senior reviewers from VC and biotech
  • Shepherd: Maria Marinova
  • Other squad members: Abhijit Kale, Ryan Spangler, Cassy Le, Adrian Matysek
  • Sourced by: Maria Marinova

Project PI

Mario D. Cordero, PhD

Principal Investigator

Department of Molecular Biology and Biochemical Engineering, Universidad Pablo de Olavide, Seville, Spain

Simple Summary

Inflammasomes are multiprotein complexes recruited and assembled upon exposure to pathogenic microbes and host danger signals, initiating the inflammatory response and activating caspase-1. However, they are also involved in the development of inflammatory disorders and the onset of age-related diseases, such as metabolic or neurodegenerative conditions.

The Principal Investigator, Mario D. Cordero, PhD, and his team showed in their preliminary results that different inflammasomes interact in the early stages of the inflammatory response, setting the hypothesis that the formation of heterocomplexes of inflammasomes is a key regulatory pathway for for pro-inflammatory mediators such as IL-1β and IL-18.

The team will explore the molecular links between the signaling mechanisms of inflammasomes to form regulatory heterocomplexes throughout the lifetime. They will address the formation of a coordinated macromolecular self-regulatory network and its malfunction in age-associated phenotypes and during cellular senescence as a physiological source of age-associated damage. Lastly, Cordero and his team will generate an innovative set of potential therapeutic candidates that will lay the basis for novel, long-lasting effective treatments for age-related diseases.


Aging is a natural process for all living organisms based on a progressive impairment of the physiological homeostasis at the cellular, tissue, and organismal level. The biological hallmarks of aging include inflammation. “Inflammaging” refers to the progressive accumulation of pro-inflammatory mediators during biological aging creating a low- grade sterile chronic inflammation environment. Among these mediators, IL-1β and IL-18 have been extensively studied and are directly related with the formation and activation of the inflammasomes.

Inflammasomes are multiprotein complexes responsible for the regulation of caspase-1 activation and the induction of inflammation in response to infectious microbes and host danger signals. However, inflammasomes activation in microglial cells and macrophages is associated with aging and age-related neurodegenerative diseases.

It is therefore paramount to not only understand the physiological function of inflammasomes, but to also explore ways to regulate it in order to develop short and long-term personalized treatments.


The primary objective is the development of therapeutics for age-related diseases through an in-depth study of inflammasomes, crucial agents in immunity and inflammation. To achieve this goal, the present project aims to study how the inflammasomes form heterocomplexes taking advantage of similar domains to participate in the aging process to trigger age-related diseases, and acting as sensors of danger signals associated to aging.

The investigator propose to reach these goals by using a multi-disciplinary approach including cell culture, molecular biology, structural biology, biophysics and animal models to specifically address the following objectives:

WP1.- Characterization of the heterocomplexes involved in life and aging.

WP2.- Mechanism of interaction of the inflammasome to participate in the formation of heterocomplex structures.

WP3.- Design of novel pharmacological approaches to modulate superinflammasomes activity.

The research fills a critical gap in the current market, as there is a lack of comprehensive solutions targeting the complexities of inflammaging.

Other research predominantly focus on the development of inhibitors targeting individual inflammasomes. However, this approach often overlooks the compensatory mechanisms that can arise, diminishing long-term efficacy. With this project, the team is introducing a novel therapeutic target that can prevent age-related diseases by pioneering a new approach in the management of inflammaging.

Experimental plan and budget

Research Plan

Objective 1. Characterization of the inflammasomes macro-complexes involved in life and aging.

The investigator’s preliminary data showed an activation of NLRP1 and NLRP3 in an irradiated cellular model of senescence and animal model of accelerated aging (Figure 1). And other inflammasomes have been shown to be activated during aging such as NLRC4 or AIM-2. All this data opens the door to the involvement of multiple inflammasomes in various stages of aging and senescence with interaction between them.

Figure 1. Inflammasomes expression during aging and senescence

Objective 2. Human validation of the inflammasome heterocomplexes

In numerous human diseases where the inflammasome has been studied, multiple papers provide evidence of various activated inflammasomes. Consequently, several cardiovascular, metabolic, and neurodegenerative diseases have demonstrated elevated levels of NLRP1, NLRP3, NLRC4, or AIM2 inflammasome complexes. The investigator and his team aim to validate findings from other objectives using human samples obtained from diverse diseases.

Objective 3. Design of novel pharmacological approaches to modulate heterocomplexes

The investigator will use the structural data available on the different individual inflammasomes to perform a rational-driven search for novel modulators of heterocomplexes.

The compound will also be evaluated in vivo models of aged mice, accelerated aging model (progeroid mice) and a gain of function NLRP3 mutant associated with neonatal onset multisystem inflammatory disease (NOMID) in mice. The investigator will analyse survival (Kaplan–Meier survival curves) bodyweight, food and water consumption, cardiometabolic profile (electrocardiographic examination, glucose tolerance and insulin response, Leptin, adiponectin and serum biomarkers) and inflammatory profile (Cytokine Array / Chemokine Array 42-Plex with IL-18 (HD42)), histological of liver, heart, kidney, lung, muscle, ovary and skin from male and female mice.


Moreover, Cordero’s laboratory has developed a compound with the potential to inhibit both NLRP1 and NLRP3, which are two major inflammasomes involved in distinct inflammatory pathways. Simultaneously targeting both could offer a broader spectrum of action, impacting various inflammatory conditions or diseases. Preliminary data indicate that NEK7 is an essential protein in the activation of both NLRP1 and NLRP3 (Figure 2). Cordero’s initial compound, C3, demonstrates significant effects in inhibiting the interaction of NEK7 with NLRP1 and NLRP3.

Figure 2. Experimental confirmation of an inflammasome heterocomplex by Native PAGE and NMR.



Postdoctoral researcher: 25.500/year (2 years)

Total (2 years): USD$ 51 000

The investigator requests hiring of a postdoctoral researcher to work on the objectives of the project and give stability to the research team of Dr. Mario D. Cordero.
The rest of the requested budget is for consumables, reagents and animal facility and mice cost, central services, publication expenses and registration fees for congresses and trips for the dissemination of the results of the project.

Consumables Year 1 Year 2
Animal costs (mice, animal facility) 8 800 8 800
Chemical synthesis 8 000
Antibodies 6 000 4 500
Reagents 10 000 9 000
Elisa Kits 3 500 3 000
X-ray facility 800 1 000
Microscopy facilities 3 500 3 000
Western blots assays 6 000 5 000
Statistical assistance 1 500
Publications 4 000 4 000
Travel allowances
Registration, accommodation, transport 5 000 5 000
Sub-total 47 600 52 800
Sub-total execution costs and travel allowance: USD$ 100 400
IN TOTAL USD$ 151 400

VitaDAO Funding Terms

Sponsored Development Agreement or Decentralised Tech Transfer model (reference: VDP-45) with terms to be agreed with TTO. Discussion ongoing.

Relevance to longevity

This project is highly relevant to longevity due to its focus on inflammasomes, which are multiprotein complexes involved in both the inflammatory response and the onset of age-related diseases like metabolic and neurodegenerative conditions. The Principal Investigator, Mario D. Cordero, and his team aim to explore how different inflammasomes interact in early inflammatory stages and hypothesize that their formation into heterocomplexes is a crucial regulatory pathway for these proteins.

Understanding these molecular links and the formation of regulatory heterocomplexes throughout a lifespan could uncover insights into age-related damage and cellular senescence. By investigating the malfunction of these complexes in age-associated phenotypes, the team hopes to generate potential therapeutic candidates for treating age-related diseases more effectively.


Dr. Mario D. Cordero, Principal Investigator

Dr. Antonio Astorza, Postdoctoral Researcher

Dr. Juan Miguel Suarez, Postdoctoral Researcher

Inés Muela, Predoctoral Student

Juan A. Pedregal, Agent of the Intellectual Property office of the Universidad Pablo de Olvide


Innovative Approach: The focus on studying inflammasome heterocomplexes during aging is innovative, offering a unique perspective on the inflammatory processes associated with aging.

Preliminary Compound Development: Having a family of compounds with inhibitory potential against a heterocomplex formed by NLRP1/NLRP3 provides a strong starting point for drug development.

Experienced Research Team: The expertise of the Principal Investigator and his team, backed by previous research demonstrating the roles of inflammasomes in various aspects of aging, serves as a strong foundation for this project.


Complexity of Inflammasome Interactions: Understanding the interactions between different inflammasomes can be highly complex. Validating specific heterocomplexes associated with aging and diseases might face challenges due to the intricate nature of these molecular interactions.

Translation to Therapeutics: While the preliminary compounds show promise, translating them into effective therapeutics for human use might face hurdles such as toxicity, off-target effects, or limited efficacy in vivo.

Long-Term Effects and Safety: Given that treatments for age-related diseases might require long-term administration, ensuring the safety and efficacy of inhibitors over extended periods, especially in aging populations, could pose challenges.

Senior Review Summary

Total senior review conviction score: 3

Reviewer 1: Score - 2
The project is highly relevant to longevity for various aging diseases, inflammation, and metabolic diseases. The team is capable of conducting their proposed experiments and consists of experts in the inflammasome field. However, the team lacks any experts in early-stage drug development.
The novelty is also unclear due to the development stage, and many groups/companies targeting the inflammasome. Data is lacking and/or of low quality.

Reviewer 2: Score - 3
The team, scientifically qualified to run this project, currently lacks clarity on their experience in transitioning this project from an academic setting to a startup. Elaboration is needed for the work packages seeking funding.
Successful outcomes from this project could yield applications in ovarian aging, chronic diseases, and various age-related diseases, with very high commercial potential and impact. However, competition in this space remains challenging due to its importance.
The approach lacks novelty, given other ongoing efforts by multiple research groups and companies developing NLRP1 and NLRP3 inhibitors at clinical or more advanced stages. However, planned experiments hold potential for intellectual property generation.
The project’s alignment with the VitaDAO longevity and healthspan extension mission is clear.

Reviewer 3: Score - 3
I feel this is well aligned with VitaDAO’s mission, provided it is negotiated to be an IP-NFT deal or similar. If the hypothesis that a reduction in inflammation reduces the negative impacts of aging holds up, it would be the first aging therapeutic to target this if successfully brought to market.
However there is no translational team in place to bring a drug to the clinic if interesting data is uncovered.

Reviewer 4: Score - 3
The targets and cytokines discussed that are part of the inflammasome pathway, such as IL-18, are well-characterized and employed in other therapeutic strategies. The screening project seems viable but it is just that - a screening project.
I do think the therapeutic area focus and general area of research are relevant for longevity and, therefore, rated this a bit higher.

Reviewer 5: Score - 4
The project still has a long way to go before commercialization. Right now, Dr. Cordero and his team are good enough but should start reaching out to partners and advisors.
The project is viable, but I do not think the data is sufficient, which is quite normal at its current stage.
NLRP1 & NLRP3 have a broad range of disease indications such as cardiovascular diseases & Alzheimer’s disease.

*Please note: There are only $125k remaining in the treasury for IP-NFT deals at this moment.

  • Agree
  • Revisions Requested (Detail in Comments)
  • Disagree
0 voters

The inflammasome field is exceptionally competitive. These data fail to come anywhere close to even the medium players in the field.

Objective 1
Showing NLR expression is not sufficient. IL-1b, IL-18 secretion, Casp1, Gasdermin D cleavage, and cell death with and without blockade of the inflammasome pathway using established inhibitors (hello MCC950) are all standard and necessary data before you can claim a functional inflammasome.

Also, I would be surprised if aging mice have not yet been crossed with the NLRP3 knockouts and the Casp1/11 knockouts. So I would expect this aim is already done, with higher rigor.

Objective 2
Inflammasome activation in human tissues is well established in all of the human diseases listed. There is no clear question or hypothesis here.

Objective 3
This is what everyone is doing. I see nothing in the data to suggest this project will find a new inhibitor before anyone else does. Worse, there are no plans to test the function of the compound to see 1) if it impacts inflammasome activation and 2) if it can succeed where MCC950, YVAD and all of the other inflammasome inhibitors have failed.

NEK7 was published as being essential for NLRP3 activation 7 years ago.

Targeting NEK7 is a reasonable strategy, but Fig 2 fails to show that there is a working inhibitor. Need to see functional impacts, not protein levels without actin controls. Assuming the actin in the SDS-PAGE in Fig 2 is consistent, the phenotype appears to target inflammasome priming, not inflammasome activation. That means the compound does NOT target NEK7 nor the inflammasome, but instead targets the induction step mediated in that experiment by LPS. Or it has many off-target effects.

Also, using the dominant active NLRP3 mouse model is a poor choice if the compound is supposed to work upstream of NLRP3 activation.

If there were solid preliminary data showing the proposed NEK7 inhibitor interfered with NLRP3 inflammasome activation (NOT priming), that part of the project would be worth considering. There would need to be a reasonable plan to confirm the mechanism, show that it works in vivo, PK, and tox studies.


Thank you, I’ll pass on the comments and questions to the PI to address.
In our talks with the researcher we have mainly focused on objective 3. and while the details are still being negotiated, this is likely the direction the project will go if it passes to further stages.
He has already provided some useful data on the inhibitor in question, including in vivo data that we can’t show here because it’s under NDA.

Would this be an IP-NFT or IPT project?

Yes, IP-NFT/potential spinout

1 Like

The experiments are very early, most lack error bars and/or are have a very low number of animals or replicates. Western blots of n=1/2 are nearly meaningless. The inventors lead compound (C3) was compared to an existing NLRP3 inhibitor (MCC950). They were comparable in efficacy. Animal studies were not compared to MCC950, and again low N. MCC950 was also discontinue due to liver toxicity in patients, which could completely invalidate the target all together.

Aquilion, Dartmouth, Neumora, have specific NLRP3 inhibitors, NodThera, Cisen Pharma, Evenda Biosciences, AC Immune, Ionis…. many many others have NLRP3 direct/indirect inhibitors.

MCC950, 3,4-Methylenedioxy-β-nitrostyrene (MNS), CY-09, OLT1177, and Oridonin are all direct inhibitors of NLRP3 and some are in clinical trials for various conditions including neurological diseases.

Parthenolide, VX-740, VX-765, Bay 11-7082, β-Hydroxybutyrate are all indirect inhibitors NLRP3/inflammasome.

It’s unclear how this project has anything that’s better than what’s in the clinic or late stage. Also, what has failed? Why? Efficacy? Safety? Some NLPR3 inhibitors have had significant tox (liver). The inventors show tox with a couple mice, but don’t compare it to MCC950 or any positive control.

The inventors say they have identified compounds. It’s unclear if these are novel or have been previously described. It’s unclear if any type of patent analysis has been performed.

Generally it lacks a convincible dataset, many other drugs already in development, and lacks any viable IP (comp of matter, methods are meaningless). Most of the plan is to explore the mechanism of inflammasomes, not develop commercialize drug assets. The inventors should come back to Vitadao after they have completed Aim 3 of their proposal as this would be a better stage to adequately asses its commercial potential.


Thanks Anthony, I’m adding Mario’s responses around these questions here for visibility:

Thank you very much for these comments about our project. We would like
to note that our preliminary data from the project are preliminary in that time point
when we applied the grant. We have been working to get more data, for example,
using another NLRP3 inhibitor as the OLT1177 which has been tested in clinical trial
with good tolerability and efficacy avoing the hepatic toxicity observed with MCC950.
To this respect, our findings using C3 vs OLT1177 in two models: Progeroid mice and
chemotherapy-dependent ovarian aging showed that C3 was more efficient compared
with OLT1177. This point show that in all our experiments we always include a
specific inhibitor of NLRP3 to compare with our drugs. On the other hand, we would
like to clarify that we have included preliminary data but the project is for demonstrate
how important is the collaboration between the inflammasome during aging and find
drugs to induce the inhibition of dual inflammasomes or more. So, this preliminary
data are only to show interesting findings about the viability of our proposal.

Our proposal show preliminary findings about the protein:protein
interaction between the inflammasomes, for examples NLRP1:NLRP3. Actually, we
have more findings showing this interaction using mass spec and CryoEM which
shouw that the inhibition of only NLRP3 could reduce NLRP3 but also induce a
compensatory effect of, for example NLRP1. A short term treatment using NLRP3
inhibitor could be efficient for several diseases but in many cases, for example, age
related diseases the treatment are for long time. Long term treatment using NLRP3
inhibitors are not been studied in animals or in vitro. We have in vitro experiments in
which NLRP1 show overexpression associated to the use of MCC950 for in vitro long
term treatment and losing efficiency. So, our proposal would improve the other
strategies from other companies supporting the dual inhibitors.
Finally, we would like to comment that we don´t know were can see the reviewer the
number of mice used in our in vivo experiments, for example, we have not tested the
toxicity using only two mice. All our animal studies use more tan 6-8 mice but we
show preliminary or representative data.