Item 1A. Risk Factors 21
Item 1B. Unresolved Staff Comments 56
Item 1C. Cyber Disclosure 56
Item 2. Description of Property 57
Item 3. Legal Proceedings 57
Item 4. Mine Safety Disclosure 57
Item 5. Market for Common Equity and Related Stockholder Matters 57
Item 6. Reserved 58
Item 7A. Quantitative and Qualitative Disclosures About Market Risk 65
Item 8. Financial Statements 65
Item 9A. Controls and Procedures 65
Item 9B. Other Information 66
Item 9C. Disclosure Regarding Foreign Jurisdictions that Prevent Inspections 66
PART III 66
Item 10. Directors, Executive Officers and Corporate Governance 66
Item 11. Executive Compensation 71
Item 14. Principal Accountant Fees and Services 85
Item 15. Exhibits, Financial Statement Schedules 86
Signatures 88
Unless
we have indicated otherwise, or the context otherwise requires, references in this Report to “MIRA,” the “Company,”
“we,” “us” and “our” or similar terms refer to MIRA Pharmaceuticals, Inc., a Florida corporation.
From
time to time, we may use our website, our Facebook page at https://www.facebook.com/people/MIRA-Pharmaceuticals-Inc/100087641460083,
our Twitter at https://twitter.com/PharmaMIRA and on our LinkedIn account at www.linkedin.com/company/mira-pharmaceuticals-inc to distribute
material information. Our financial and other material information is routinely posted to and accessible on the Investors section of
our website, available at www.mirapharmaceuticals.com. Investors are encouraged to review the Investors section of our website because
we may post material information on that site that is not otherwise disseminated by us. However, information that is contained in and
can be accessed through our website, our Facebook page, our Twitter posts and our LinkedIn posts are not incorporated into, and does
not form a part of, this Report.
i
CAUTIONARY
NOTE REGARDING FORWARD-LOOKING STATEMENTS
This
Annual Report on Form 10-K (this “Report”) contains forward-looking statements (as defined in Section 27A of the Securities
Act of 1933, as amended, and Section 21E of the Securities Exchange Act of 1934, as amended (the “Exchange Act) that reflect our
current expectations and views of future events. In some cases, you can identify forward-looking statements by terms such as “may,”
“will,” “should,” “expect,” “plan,” “anticipate,” “could,” “intend,”
“target,” “project,” “contemplate,” “believe,” “estimate,” “predict,”
“potential”, or “continue” or the negative of these terms or other similar expressions. In particular, statements
about our pre-clinical and clinical trials and expectations regarding such trials, the markets in which we operate, including growth
of such markets, and our expectations, beliefs, plans, strategies, objectives, prospects, assumptions, or future events or performance
contained in this Report generally under the headings “Risk Factors,” “Management’s Discussion and Analysis of
Financial Condition and Results of Operations” and “Business” are forward-looking statements.
We
have based these forward-looking statements on our current expectations, assumptions, estimates and projections. While we believe these
expectations, assumptions, estimates, and projections are reasonable, such forward-looking statements are only predictions and involve
known and unknown risks and uncertainties, many of which are beyond our control. These and other important factors, including those discussed
in this Report under the headings “Risk Factors,” “Management’s Discussion and Analysis of Financial Condition
and Results of Operations” and “Business,” may cause our actual results, performance, or achievements to differ materially
from any future results, performance or achievements expressed or implied by these forward-looking statements, or could affect our share
price. Important factors that could cause actual results or events to differ materially from those expressed in forward-looking statements
include, but are not limited to, the following
● the timing of anticipated regulatory filings;
● the timing and availability of data from our clinical trials;
● our ability to recruit and enroll suitable patients in our clinical trials;
● the pricing and reimbursement of our product candidates, if approved;
● developments relating to our competitors and our industry;
Given
the risks and uncertainties set forth in this Report, you are cautioned not to place undue reliance on such forward-looking statements.
The forward-looking statements contained in this Report are not guarantees of future performance and our actual results of operations,
financial condition, and liquidity, and the development of the industry in which we operate may differ materially from the forward-looking
statements contained in this Report. In addition, even if our results of operations, financial condition and liquidity, and events in
the industry in which we operate, are consistent with the forward-looking statements contained in this Report, they may not be predictive
of results or developments in future periods.
Any
forward-looking statement that we make in this Report speaks only as of the date of such statement. Except as required by federal securities
laws, we do not undertake any obligation to update or revise, or to publicly announce any update or revision to, any of the forward-looking
statements, whether as a result of new information, future events or otherwise, after the date of this Report.
Summary
of Principal Risks
Our
business is subject to numerous risks and uncertainties that represent challenges that we face in connection with the implementation
of our strategy and the growth of our business. In particular, the following are the principal risks which could cause a decline in the
price of shares of our common stock:
PART
I
ITEM
1. Description of Business
Overview
MIRA
Pharmaceuticals, Inc. (NASDAQ: MIRA) is a clinical-stage pharmaceutical development company advancing two neuroscience programs targeting
neurologic and neuropsychiatric disorders. The company holds exclusive rights in the U.S., Canada, and Mexico for Ketamir-2 and MIRA-55,
two novel drug candidates designed to address unmet medical needs in pain management, depression, PTSD and cognitive function.
Ketamir-2:
A Novel Oral Ketamine Analog
Ketamir-2
is a patent-pending, oral ketamine analog currently being evaluated for the treatment of diabetic neuropathy and is in an ongoing Phase
I clinical trial. The compound is designed to overcome the limitations of existing ketamine-based treatments, offering the potential
for enhanced safety, improved tolerability, and better oral bioavailability. If approved, Ketamir-2 could provide a safer, more effective
option for patients suffering from neuropathic pain, treatment-resistant depression (TRD), major depressive disorder with suicidal ideation
(MDD-SI), and post-traumatic stress disorder (PTSD), with the possibility of additional Phase I studies exploring these indications.
Neuropathic
pain is a complex, chronic condition resulting from damage or dysfunction in the nervous system, leading to abnormal sensations or pain
responses. Common symptoms include burning, coldness, “pins and needles” sensations, numbness, itching, and sudden electric
shock-like pain. Neuropathic pain can arise from metabolic disorders (such as diabetic neuropathy), viral infections (like post-herpetic
neuralgia), autoimmune diseases, chemotherapy-induced neuropathy, traumatic nerve injury, stroke, or spinal cord injury.
Existing
treatment options include antidepressants (tricyclic antidepressants and serotonin-norepinephrine reuptake inhibitors), anticonvulsants
(gabapentin and pregabalin), topical agents (lidocaine patches and capsaicin), and opioids. However, these therapies often provide incomplete
relief, have significant side effects, and, in the case of opioids, carry a high risk of addiction. Ketamine has demonstrated efficacy
for neuropathic pain, but its poor oral bioavailability and high risk of side effects have limited its widespread use. Ketamir-2 aims
to address these challenges, offering a treatment with fewer side effects, lower abuse potential, and more convenient oral dosing.
MIRA-55:
A Novel Oral Pharmaceutical Marijuana Analog
MIRA-55
is a preclinical-stage investigational drug designed to support cognitive function and enhance memory. It is currently being evaluated
in preclinical studies for its potential benefits in neuropsychiatric, inflammatory, and neurologic disorders. If approved, MIRA-55 could
provide a breakthrough treatment for patients experiencing cognitive decline, including those with early neurodegenerative conditions.
Regulatory
and DEA Classification
The
U.S. Drug Enforcement Administration (DEA) has completed its scientific review of both Ketamir-2 and MIRA-55, determining that neither
compound will be classified as a controlled substance or listed chemical under the Controlled Substances Act and its governing regulations.
This regulatory distinction significantly enhances their commercial and clinical viability, removing potential barriers associated with
controlled substances and positioning both compounds for streamlined clinical development and commercialization.
MIRA
Pharmaceuticals remains committed to advancing Ketamir-2 and MIRA-55 through clinical development with a focus on addressing major unmet
medical needs and improving patient outcomes.
Preclinical
Studies and Pharmacology of Ketamir-2
MIRA
has conducted extensive preclinical studies to characterize the pharmacological profile, safety, and therapeutic potential of Ketamir-2,
an investigational compound targeting neuropathic pain, treatment-resistant depression (TRD), major depressive disorder with suicidal
ideation (MDD-SI), and post-traumatic stress disorder (PTSD). These studies, performed in accordance with ICH and FDA regulatory guidelines,
include in vitro and in vivo assessments to evaluate receptor binding, efficacy, selectivity, pharmacokinetics, metabolism, general pharmacology
and toxicology.
Mechanism
of Action and Receptor Selectivity
Ketamir-2
has been identified as a low-affinity NMDA receptor antagonist, selectively binding to the PCP-site with an IC50 of approximately 100
μM. Its primary metabolite, Nor-Ketamir, also binds to the PCP-site and also does not exhibit affinity for NMDA site, with an IC50
of approximately 300 μM. This targeted receptor interaction differentiates Ketamir-2 from ketamine, which exhibits broader receptor
binding, including opioid and monoaminergic receptors, and binds to NMDA receptors with significantly higher affinity (0.5–1 μM).
Efficacy
in Animal Models
The
therapeutic potential of Ketamir-2 was evaluated in preclinical neuropathic pain and depression models, demonstrating significant effects:
Recent
Developments
Executive
Incentive Compensation Program
On
March 26, 2025, the compensation committee (the “Committee”) of MIRA Pharmaceuticals, Inc. (the “Company”) adopted
the Company’s Executive Incentive Compensation Plan (the “EICP”) for Erez Aminov, its Chairman and Chief Executive
Officer. The EICP was approved by the Committee following greater presentation of the EICP to the board of directors of the Company (the
“Board”). The specific terms of the EICP were prepared by the Board’s independent compensation consultant.
Under
the EICP, Mr. Aminov is eligible to receive an annual target bonus of $242,500, with a maximum bonus of up to $485,000 (the “Annual
Target Bonus”). The Annual Target Bonus is equally weighted (though as adjusted as appropriate) based on the following three components:
(i) achievement of clinical milestones for the Company’s drug candidates, (ii) entering into certain strategic partnerships, and
(iii) achieving capital raise milestones. Each component under the 2025 Program may be achieved and a corresponding payout made independent
of the other components, but only after such component meets the minimum threshold of $40,017 is reached before any bonus payments will
be made.
Under
the EICP, Mr. Aminov will be eligible for certain long-term awards of up to 500,000 performance-based units of the Company’s common
stock, par value $0.001 upon the Company achieving specified milestones based upon the Company reaching certain market capitalization
values and the progress of the Company’s drug candidates.
Mr.
Aminov will also be entitled to an amount equal to 3% of the total value of any mergers and acquisition or strategic transaction completed
by the Company.
All
awards under the EICP are subject to the approval of the Board and the Committee. Furthermore, the Board and the Committee, each in its
sole discretion, generally retain the right to amend, supplement, supersede or cancel any awards under the EICP for any reason, and reserve
the right to determine whether and when to pay out any bonus amounts pursuant to or outside of the EICP, regardless of the achievement
of the performance targets.
In
addition, the Committee approved an increase of Mr. Aminov’s salary to $485,000 effective April 1st.
Binding Letter of Intent with SKNY Pharmaceuticals
Inc.
On March 19, 2025, the Company
entered into a binding letter of intent (the “LOI”) with SKNY Pharmaceuticals, Inc. (“SKNY”), a privately held
Delaware corporation, to acquire SKNY through a stock exchange transaction (the “SKNY Acquisition”). The acquisition will
bring SKNY-1, a novel oral drug candidate targeting weight loss and smoking cessation—two of the leading causes of preventable death—into
MIRA’s development pipeline. As part of the agreement, SKNY will provide a $5 million capital infusion in cash or cash equivalents,
further strengthening MIRA’s financial position and supporting future growth initiatives.
SKNY holds exclusive rights to
its compounds in the United States, Canada, and Mexico. Under the terms of the LOI, SKNY will merge into the Company through a stock exchange,
with each outstanding share of SKNY’s common stock being exchanged for shares of MIRA’s common stock. The exact exchange ratio
will be determined by an independent third-party valuation firm (the “Independent Valuator”) based on the relative values
of both companies. The completion of the SKNY Acquisition is contingent upon the Independent Valuator determining that SKNY’s valuation
is at least equal to or greater than that of the Company. The SKNY Acquisition will be subject to shareholder approval of both companies.
Safety
and Toxicology Studies
Cardiovascular,
CNS, and Respiratory Safety
A
GLP-compliant safety pharmacology program evaluated the effects of Ketamir-2 on the cardiovascular, central nervous system (CNS), and
respiratory function using standard nonclinical models.
Toxicology
and Dose Tolerability
Comprehensive
toxicology studies were conducted in Sprague Dawley rats and Beagle dogs to assess systemic exposure, metabolism, and dose tolerability.
Pharmacokinetics
and Metabolism
A
series of in vivo pharmacokinetics (PK) and toxicokinetic (TK) studies assessed the absorption, metabolism, and systemic exposure of
Ketamir-2.
Ketamir-2
Clinical Development Program
The
clinical development program for Ketamir-2 follows a structured, multi-phase approach, beginning with IND-enabling studies and progressing
through Phase I and Phase IIa clinical trials. The objective is to evaluate the safety, efficacy, and optimal use of Ketamir-2 for the
treatment of diabetic neuropathy, with a focus on patient safety and regulatory compliance. If successful, Ketamir-2 could provide a
novel treatment option that addresses the limitations of existing therapies for neuropathic pain management.
IND-Enabling
Studies and Regulatory Progress
The
development pathway began with the completion of IND-enabling studies, including pharmacokinetics, pharmacodynamics, toxicology, and
safety pharmacology assessments in compliance with FDA regulatory standards. Following the submission of an Investigational New Drug
(IND) application for diabetic neuropathy to the FDA in December 2024, the agency provided feedback requiring the completion of a neurotoxicity
study before proceeding with human dosing in the United States. This study is currently ongoing, and we expect it to be completed by
May 2025.
While
awaiting the completion of the neurotoxicity study, we have received approval from the Israeli Ministry of Health and Institutional Review
Board (IRB) to initiate our Phase I studies. The study will be conducted at the Clinical Pharmacology Unit, Hadassah Medical Center in
Jerusalem, Israel. We plan to begin Phase I clinical trials in Israel in the first quarter of 2025, followed by Phase IIa studies in diabetic
neuropathy patients to be initiated in the fourth quarter of 2025.
Phase
I: Safety and Dosage Determination in Healthy Volunteers
Study
Design
● Randomized, double-blind, placebo-controlled trial.
● Primary objective: Assess safety and tolerability of Ketamir-2.
Participant
Selection
● Healthy adult volunteers representing a diverse demographic population.
Dosing
and Administration
● Continuous monitoring for adverse effects.
Outcome
Measures
Phase
IIa: Dose, Tolerability, and Early Efficacy in Diabetic Neuropathy
Following
the completion of Phase I, Phase IIa clinical trials will focus on diabetic neuropathy patients. This phase will evaluate the optimal
dose and tolerability while also collecting preliminary efficacy data.
Study
Design
● Randomized, controlled trial in patients with diabetic neuropathy.
● Primary objective: Identify optimal dose and tolerability.
● Secondary objective: Assess early efficacy outcomes.
Participant
Selection
Dosing
Regimen
● Dose range based on Phase I findings.
● Flexible or fixed-dose regimens, depending on safety and tolerability data.
Outcome
Measures
● Efficacy assessments: Several standardized pain measures.
Clinical
Manufacturing and Drug Supply
Recipharm
Israel Ltd., a leading global contract development and manufacturing organization (CDMO), has successfully completed the upscaling, development,
and GMP manufacturing for clinical and preclinical supplies of Ketamir-2.
MIRA-55:
A Novel Oral THC Analog
Our
objective is to develop and commercialize new treatment options for neuropsychiatric, inflammatory, and neurologic diseases and disorders.
Cannabinoids are a class of chemical compounds that are naturally occurring and are primarily found in cannabis plant extracts. The two
major cannabinoids found in cannabis plant extracts include tetrahydrocannabinol, a compound that is the main psychoactive ingredient
of cannabis (or THC) and cannabidiol, the second most prevalent active ingredient in cannabis which does not have psychoactive properties
(or CBD). These compounds bind to CB1 and CB2 cannabinoid receptors, which are found throughout the body. Specifically, CB1 receptors
are concentrated in the central nervous system (or CNS), while CB2 receptors are found mostly in peripheral organs and are associated
with the immune system. When the chemical compounds bind to these cannabinoid receptors, the process elicits certain physiological responses.
Physiological responses to cannabinoids may vary among individuals. Some of the effects of cannabinoids have been shown to impact nervous
system functions, immune responses, muscular motor functions, gastrointestinal maintenance, blood sugar management, and the integrity
of ocular functions. Based on pre-clinical testing, our product candidate, MIRA-55, appears to have a strong selectivity for CB2 versus
CB1, and is designed to minimize the risk of psychoactive adverse events associated with CB1 activation.
Mechanism
of Action of MIRA-55
We
believe that the effects of MIRA-55 at the cannabinoid receptors CB1 and CB2 is predicted to account for most of its potential therapeutic
effects, especially as it relates to its anti-inflammatory properties. For example, the difference in the dose-response effects of MIRA-55
compared with THC on CB1 receptors appears to coincide with its improved therapeutic profile. If approved by the FDA, MIRA-55 may potentially
provide therapeutic effects and enhanced cognition and memory.
THC
has been demonstrated to have biphasic physiological effects (meaning effects in two phases), which have been described for over 40 years:
at low levels THC has positive effects while high doses cause the opposite, undesirable symptoms. Examples of biphasic effects at low
versus high levels of THC include the anti-anxiety versus pro-anxiety effects, respectively. Through pre-clinical test, we obtained the
following dose-response effects for MIRA-55 and THC at the CB1 receptor (see below). In contrast to THC, which displays an initial maximally
stimulatory and then inhibitory response at CB1, MIRA-55 appears to act as a monophasic partial agonist (meaning it has a lower intrinsic
activity than full agonists) in that it creates a stimulation throughout its dose range, achieving a moderate activation of the CB1 even
at high doses. We believe that this accounts for the potential broad therapeutic efficacy of MIRA-55 and the observed absence of negative
symptoms even at maximal doses of the drug.
Figure
1: Compound activity with the selected GPCR Biosensor Assays: THC vs MIRA-55 agonist activity at the CB1 Receptor.
In
pharmacology, “efficacy” or “Emax” refers to the maximum response that can be achieved with a drug or agent.
It represents the extent or magnitude of the response produced by the drug once it has bound to its target, typically referred to as
a receptor. The binding between a drug and its receptor is characterized by affinity, which quantifies the strength of their interaction.
Efficacy, however, assesses the action or effect of the drug following binding to the receptor.
The
dose-response curve is a commonly used graph in pharmacology that depicts the relationship between the effect of a drug and its dosage.
The X-axis represents the increasing doses of the drug, while the Y-axis represents the response produced by the drug. In the case of
the figure above, the term “% Efficacy” on the Y-axis refers to the maximum response that can be achieved with the agonist
(MIRA-55 or THC) in relation to its ability to activate GPCR receptors (specifically CB1 receptors). GPCRs are G-protein-coupled receptors
that form a large group of proteins which are expressed on the cell surface of eukaryotic cells to detect molecules outside the cell
and activate cellular response.
The
data presented in the figure above has been normalized to the maximal and minimal responses observed in the presence of a control compound
and vehicle, respectively. This normalization allows for a standardized comparison of the agonist’s efficacy.
MIRA-55
Preclinical Developments and Studies
As
of the date of this report, we have completed several preclinical studies of MIRA-55, including, but not limited to, radio-ligand binding
assays, the elevated plus maze (EPM) model of anxiety, and hot plate model thermal sensitivity testing.
We
have studied the effects of acute administration of MIRA-55 on anxiety-related phenotypes in mice to model human conditions. An intraperitoneal
injection of placebo (e.g., saline) or MIRA-55 (50 mg/kg treatment) was administered to C57Bl/6 mice (n=5/group) that were 8-12 weeks
old. Thirty minutes following injection, mice were tested in anxiety-related measures using the Elevated Plus Maze (EPM). The EPM is
a widely used preclinical behavioral assay for rodents and has been validated to assess the anti-anxiety effects of pharmacological agents.
If determined and approved by the FDA or other regulatory agencies, MIRA-55 appears to have anti-anxiety effects at doses that lacked
side effects of sedation or intoxication in mice. The EPM measures anxiety in rodents as a screening test for putative anxiolytic compounds
and as a general research tool in neurobiological anxiety research, such as Generalized Anxiety Disorder (GAD) or Post-Traumatic Stress
Disorder (PTSD). The model is based on the animal’s aversion to open spaces, which are present in the open arms of the maze. Anti-anxiety
effects of test agents are demonstrated by an increase in the percentage of time spent in the open arms with treatment compared to placebo.
The total distance traveled is a measure of the overall level of arousal and mobility of the mice undergoing testing on the EPM and is
used to rule out any sedating or intoxicating effects of the test agent.
Preclinical
studies have also shown the potential of MIRA-55 for relieving pain. Several clinically approved pharmacological agents used to treat
pain, including opioids, have been demonstrated to delay or ameliorate the onset of heat sensitivity upon paw exposure of mice to heat.
Thirty minutes after treatment with either a placebo (control) or MIRA-55, mice were placed on a heated plate to measure the time it
took for each mouse to lift its paw in response to the mild pain they felt from the heat. Mice treated with pain-alleviating drugs took
significantly longer to become bothered by the heat and to lift their paws. Similarly, mice treated with MIRA-55 statistically took significantly
more time to lift their legs, indicating MIRA-55’s potential effectiveness as a possible treatment for pain in this model. If approved
by the FDA, MIRA-55 may potentially provide therapeutic effects for pain control.
MIRA-55
is a CB2 agonist, which may also be an optimal treatment for neurodegenerative diseases associated with neuroinflammation caused by microglial
activation. CB2 agonism has been shown in preclinical studies to regulate neuroinflammatory processes, reducing the neuronal damage characteristic
of degeneration. We believe there may be a strong rationale for CB2 agonism in neurodegenerative diseases, given increased CB2 expression
in patients with these diseases as well as preliminary results from animal models. We see potential for a potent CB2 agonist to treat
a range of neurodegenerative diseases. MIRA-55, through its robust activity at CB2 compared to CB1, was designed to minimize the risk
of psychotropic adverse events associated with CB1 activation. If approved by the FDA, MIRA-55 may potentially provide therapeutic effects
for neurodegenerative and neuroinflammatory illnesses.
Our
preclinical development program for MIRA-55 has included a variety of testing. Summarized below are the tests we have completed. Our
interpretation of results derived from preclinical data, or our conclusions based on preclinical data, may prove inaccurate and are not
necessarily predictive indicators of future results. See the section below titled “Research and Testing to Date - MIRA-55”
for more information.
In
addition to the studies completed, we have ongoing preclinical studies evaluating the effects of MIRA-55 on memory and cognition in animal
models. These studies aim to further assess MIRA-55’s potential for enhancing cognitive function and addressing deficits associated
with neurodegenerative and neuropsychiatric disorders.
Our
MIRA-55 Clinical Development Program
Following
the preclinical development plan outlined above, we intend to advance MIRA-55 toward an Investigational New Drug (IND) application submission
to the FDA for the treatment of cognitive decline in elderly patients. This patient population represents a significant unmet medical
need, and we believe MIRA-55 has the potential to provide therapeutic benefits in this area.
The
timing of our IND submission will be based on the completion of ongoing preclinical studies, regulatory discussions, and overall development
progress. If the FDA allows the program to proceed, a Phase I trial would be expected to begin following the standard regulatory review
period post-IND submission.
All
development plans are subject to FDA review and acceptance. As discussions with the FDA progress, we may refine our timeline for clinical
trials and regulatory submissions as appropriate. Given the inherent uncertainties in drug development, there is no guarantee that our
clinical development activities will align with anticipated timelines.
Market
Opportunity and Competitive Advantage of Ketamir-2 in Neuropathic Pain Treatment
Market
Opportunity
Neuropathic
pain is a significant and growing health concern in the United States, affecting approximately 10% of adults (PMC5677393). The
North American neuropathic pain market was valued at approximately $2.60 billion in 2022 and is projected to reach $5.20 billion by 2030,
growing at a 9.0% compound annual growth rate (CAGR) during this period (Data Bridge Market Research). This growth is driven by an aging
population, increasing prevalence of diabetes, chemotherapy-induced neuropathy, and post-surgical nerve injuries.
Despite
the high prevalence and economic burden, current treatment options remain limited, often ineffective, or associated with significant
side effects and safety concerns. Standard treatments include antidepressants (tricyclic antidepressants, serotonin-norepinephrine reuptake
inhibitors), anticonvulsants (gabapentin, pregabalin), opioids, and topical agents. However, these medications often provide incomplete
pain relief, require long-term use, and come with risks such as sedation, cognitive impairment, addiction, and withdrawal symptoms.
The
strong demand for innovative, effective, and safer neuropathic pain treatments presents a major market opportunity for Ketamir-2, a novel
investigational therapy designed to address these critical treatment gaps.
Competitive
Advantage of Ketamir-2
Ketamir-2
has the potential to transform neuropathic pain management by offering a more effective, safer, and accessible treatment compared to
existing therapies, including currently approved medications.
● Potential Advantages Over Current FDA-Approved Neuropathic Pain Treatments:
By
offering oral bioavailability, improved safety, reduced abuse potential, and a potential advantage over currently approved neuropathic
pain treatments, Ketamir-2 is positioned to become a leading treatment option in the rapidly growing neuropathic pain market, addressing
an urgent need for more effective and accessible pain management solutions.
Ketamir-2
Development Strategy
The
goal is to advance the development of Ketamir-2 as an orally administered medication with the potential for fewer side effects and greater
accessibility, free from the restrictions imposed by ketamine’s REMS. Ketamir-2 aims to address the urgent clinical need for a
rapid-acting treatment for diabetic neuropathy and treatment-resistant depression (TRD) in patients who can take the medication at home.
The
strategic focus is to establish proof of concept in human patients, demonstrating Ketamir-2’s safety, efficacy, and differentiation
from existing therapies. In parallel, the company will actively explore effective exit strategies, including strategic partnerships,
licensing opportunities, or potential acquisition pathways to maximize value and commercial potential.
MIRA-55
Development strategy
Our
goal is to develop therapeutics targeting well-characterized CB1 and CB2 receptors with optimized pharmacological properties to transform
the lives of patients with neurological diseases. Key elements of our strategy to achieve this goal include:
Ketamir-2
Preclinical
Research Findings
In
Silico Analysis of Targets of Ketamir-2 vs Ketamine
In
silico analysis, referring to computer-based techniques, has become an integral part of pharmaceutical research and development.7
This approach utilizes computational methods to analyze and predict the properties and behaviors of pharmaceutical compounds. The
use of in silico analysis is especially crucial in the early stages of drug development, as it aids in identifying potential drug targets
and elucidating differences between a new drug and its parent compound. By analyzing large datasets, such as genomic, proteomic, and
metabolomic data, researchers can predict how different compounds might interact with various biological targets. This approach helps
in understanding the mechanism of action of new drugs and can significantly reduce the time and cost associated with experimental screening.
InSilico Trials was contracted to provide a comparison between targets of Ketamir-2 vs ketamine employing their target identification
protocol. The following characterize some of the unique targets that are predicted to interact with either Ketamir-2 or ketamine, thereby
differentiating one drug from the next.
Studies
evaluating the mechanism of action of the investigational product indicate that Ketamir-2 exhibits a highly selective receptor binding
profile. Specifically, Ketamir-2 was found to be a low affinity NMDA receptor antagonist, that selectively binds to the phencyclidine
(PCP)-site with an IC50 on this receptor site of ~100 μM. In contrast, ketamine primarily works as an NMDA receptor
antagonist with affinities in the range of 0.5-1 μM.
Ketamir-2
selective target:
Additionally,
while ketamine has a broader receptor binding profile which includes opioid receptors and receptors associated with monoaminergic systems,
the receptor binding profile of Ketamir-2 and the primary metabolite of Ketamir-2 is specific to the NMDA receptor PCP-site.
Pharmacology:
Primary
pharmacology studies (in vitro and in vivo) conducted to support the advancement of Ketamir-2 are summarized in the table below. In vivo
pharmacology assessments include evaluation of Ketamir-2 using endpoint assessments post-dose administration that are traditionally utilized
to evaluate behavioral indices such as spontaneous horizontal locomotor activity, anxiety, and depression in addition to evaluation of
the therapeutic efficacy in a rodent model of neuropathic pain. While endpoint assessments pertaining to depression, anxiety, and general
locomotion are not direct evidence of an improvement in indices of neuropathic pain, discordant neuropharmacological signaling through
the NMDA receptor is a commonality within neuropsychiatric disorders such as depression and anxiety. Specifically, overactivation of
the NMDA receptor has been suggested to play a role in major depressive disorders for which anxiety is comorbid. Moreover, signaling
through the NMDA receptor plays a role in the pain transmission pathway and is thus associated with neuropathic pain. Thus, data derived
from assessments traditionally used in the evaluation of potential antidepressant therapeutics such as the forced swim test, elevated
plus maze, and open field test are directly linked to the mechanism of action of Ketamir-2.
MIRA
Pharmaceuticals has completed a series of nonclinical pharmacology studies characterizing Ketamir-2 efficacy in vitro and in
vivo in animal models. Results of in vitro assessments which evaluated the pharmacological activity of Ketamir-2 indicate
that Ketamir is a low affinity NMDA receptor antagonist that selectively binds to the PCP-site with an IC50 of ~100 μM
on this receptor site. Nor-Ketamir, the primary metabolite of Ketamir-2, selectively binds to the PCP-site with no affinity for NMDA.
The IC50 is ~300 μM. Evaluation of therapeutic efficacy of Ketamir-2 in animal models of depression, anxiety and neuropathic
pain (i.e., Chung model) indicate that treatment with Ketamir-2 results in anti-anxiolytic and anti-depressive behavior in behavioral
assessments such as the open-field test, elevated plus maze, and the forced swim test. Moreover, animals administered 30 – 300
mg/kg Ketamir-2 exhibited significant increases in mean withdrawal threshold on Day 15 and 22 (100 and 300 mg/kg only) when compared
to Day 14 post-surgery values. These ameliorative effects are observed in the absence of off-target binding/signaling from both Ketamir-2
and the primary metabolite Nor-Ketamir.
Figure
2: Effect of Ketamir-2 on neuropathic pain in male rats. Study represents the sensitivity to von-Frey filaments at 15 (blue) and 22 days
(purple) after induction of neuropathy.
Bioavailability:
MIRA
has conducted a series of informational ADME assessments typical for a novel therapeutic classified as a small molecule. The data indicate
that Ketamir-2 is permeable at a dose of 10 μM (A-B and B-A) and is not highly bound to plasma proteins in mouse, rat, dog, cynomolgus
monkey, or human plasma. Substrate interaction and metabolism studies indicate that Ketamir-2 is not a substrate of P-glycoprotein and
is metabolized in mice, rats, dogs, cynomolgus monkey, and human hepatocytes with half-lives between 3.26 – 18.3 min. This biotransformation,
as evaluated in human hepatic microsomes, is primarily the result of N-demethylation with metabolism being mediated primarily by CYP2B6
and CYP3A4
In
addition to these data, the pharmacokinetics of Ketamir-2 have been characterized in a series of in vivo PK/TK studies conducted in Sprague
Dawley rats and Beagle dogs. General PK data indicate that Ketamir-2 does cross the blood brain barrier with longer exposure times and
greater levels of nor-Ketamir present in the brain. Moreover, the PK profile of Ketamir-2 following oral administration is characterized
by rapid absorption, a short half-life, high clearance, and lower oral bioavailability.
Within
the definitive toxicology studies, in general, females had higher systemic exposure than males of both Ketamir-2 and nor-Ketamir at 100
and 300 mg/kg/day (Days 1 and 14) and at 1000 mg/kg/day (Day 1) in rats and at 100 mg/kg/day (Days 1 and 14) and at 50 mg/kg/day (Day
1) in canines. Within rats, systemic exposure increased dose proportionally in females on Day 1 and in both sexes on Day 14 but increased
more than dose proportionally in males on Day 1. In contrast, systemic exposure in canines increased in a more than dose proportional
manner in both sexes on Days 1 and 14. The metabolite to parent (nor-Ketamir/Ketamir-2) ratios in AUC0-24h ranged from 2.2
– 22 and 4.5 – 44 in rats and canines, respectively.
MIRA-55
Eurofins
DiscoverX has developed a panel of cell lines stably expressing non-tagged GPCRs that signal through cyclic adenosine monophosphate,
a messenger used for intracellular signal transduction in many different organisms (or cAMP). Hit Hunter® cAMP assays are specialized
tests that track the activation of a type of cell receptor known as GPCR. GPCRs play a crucial role in how cells respond to external
signals, and they are activated through two pathways: Gi and Gs secondary messenger signaling. These pathways are like internal communication
systems in cells that relay signals from the outside to trigger specific responses inside the cell. The assay is conducted in a straightforward,
uniform manner without the need for image-based analysis. This method uses a technology developed by DiscoverX called Enzyme Fragment
Complementation (or EFC). In EFC, fragments of an enzyme, specifically β-galactosidase (β-Gal), are brought together to become
functional only when the GPCR is activated. β-Galactosidase, the enzyme used as a functional reporter in this assay, is typically
inactive in fragmented form and becomes active when the fragments reassemble, indicating the activation of the GPCR. In this case, the
GPCR target was CB1 receptor. Compounds were tested in agonist and antagonist mode with the requested GPCR Biosensor Assays. For agonist
assays, data was normalized to the maximal and minimal response observed in the presence of control ligand and vehicle. This Eurofins
DiscoverX system was used to test THC vs MIRA-55 agonist activity at the CB1 receptor.
Unlike
CB1 receptors that mediate many of the psychotropic effects of cannabinoids on the CNS, CB2 receptors are predominantly present on cells
of the immune system. Based on preliminary results of our GPCR biosensor assays, the CB2 receptor agonistic effects of MIRA-55 are 8-fold
more potent than THC and 30-fold more potent than CBD.
The
study regarding the ability of MIRA-55 vs THC vs CBD to activate CB2Receptors and alter intracellular cAMP levels was performed by the
CRO Eurofins DiscoverX.
As
can be seen in the table below, the EC50 (i.e. concentration required to induce a half maximal response) for MIRA-55 was 8 times more
potent than THC and at least 30 times more potent that CBD-i.e. it only took 1 uM of MIRA-55 to induce the same response that required
8 uM of THC and >30 uM of CBD.
Compound Name Assay Name Assay Format Assay Target Result Type EC50 Unit
CBD cAMP Agonist CNR2/CB2 EC50 >30 uM
Table:
The foregoing measurements were performed as follows:
DiscoverX
has developed a panel of cell lines that stably express non-tagged GPCRs (G-protein coupled receptors) capable of signaling through cAMP.
The Hit Hunter® assay platform is used to investigate the functionality and response of these GPCRs.
In
the case of the CB2 receptor, which is a GPCR involved in various physiological processes and has potential therapeutic implications,
the Hit Hunter® assay can be employed to study the effects of drug agonists on CB2 receptor activity.
To
measure the half maximal response (EC50) of CB2 receptor activation by a drug agonist that leads to a decrease in cAMP levels, an alternative
approach may be required. One common method involves using forskolin, an activator of adenylate cyclase, to stimulate cAMP production.
Forskolin bypasses the GPCR signaling and directly activates adenylate cyclase, resulting in increased cAMP levels.
In
the presence of forskolin, the drug agonist at the CB2 receptor can then be tested at various concentrations to determine its ability
to inhibit the forskolin-induced cAMP production. The drug’s concentration that leads to a 50% reduction in forskolin-stimulated
cAMP levels can be considered the half maximal response or EC50.
Completed
Pre-Clinical Tests
EPM
Model of Anxiety Test
EPM
is a widely used behavioral test to assess anxiety-like behavior in rodents. Typically, rodents tend to avoid open spaces due to their
natural aversion to potentially dangerous areas. Therefore, spending more time in the open arms of the maze indicates decreased anxiety-like
behavior. Similarly, the total distance travelled can reflect general locomotor activity and exploratory behavior, which can be influenced
by the state of anxiety and the effect of drugs.
The
EPM apparatus consists of two open arms and two enclosed arms elevated above the floor. Blue Bars represent the percentage of time spent
in the open arms by mice in the placebo and drug-treated groups. Green Bars show the total distance travelled by mice in both groups
during the EPM test
The
issue of how to test the effect of MIRA-55 on cognition was complicated by the following:1) MIRA-55 has anti-anxiety (i.e. anxiolytic)
effects, 2) anxiolytics can potentially improve cognitive assessment outcomes by reducing anxiety levels that may otherwise hinder cognitive
functioning. Thus, in commonly performed tests of cognition in mice, such as novel object recognition and Morris water maze, anxiolytic
medications can indirectly result in improved performance by decreasing anxiety rather than by directly improving cognition. To separate
assessments of the impact of MIRA-55 on cognitive performance from its demonstrated anti-anxiety effects, we employed a model of context
fear conditioning wherein we dosed the mice after training. Context fear conditioning in mice is a behavioral paradigm used to measure
cognitive processes related to associative learning and memory. Associative learning, where an individual learns to associate specific
stimuli or contexts with outcomes, in this case the mice associate being in a specific chamber with receiving a mild foot shock that
occurs during training the day before testing. This process of forming associations between stimuli, actions, and consequences is involved
in numerous skills and behaviors in everyday life: it underlies learning new skills, developing habits, and acquiring knowledge through
experiences and conditioning. The use of associating the chamber with the foot shock on day one, means that when the mice are returned
to the chamber on day 2 a measure of how much freezing they do corresponds to a read out of how well they can recall the experiences
they had during training on day 1 (i.e. the greater the freezing, the better the recollection of the association between the chamber
and food shock). Since the mice are given MIRA-55 AFTER training that takes place on day 1, and only before testing on day 2, there is
no concern about the anxiolytic effects of MIRA-55 on learning during training, but rather this model tests MIRA-55’s effects on
performance only-which in this case represents memory (i.e. the ability to recognize and recall the chamber where they had previously
been shocked) and to translate that into an associated behavior (i.e. freezing). As published in the Journal of Neuropharmacology in
2023, THC and cannabis impair context fear conditioning, both when given prior to training (because of its anti-anxiety effects) and
when given prior to testing (because of its cognitive impairing effects). MIRA-55 resulted a dramatic effect on cognitive performance
in the context fear conditioning model, the percentage of time spent freezing-that is a demonstration of their memory and association-in
the mice who received MIRA-55 at a dose of 75 mg/kg was more than twice that of those who received 0 mg/kg=placebo (i.e. 55% vs 25%,
p<0.0001). Thus, MIRA-55 doubled the cognitive performance of the mice compared to placebo. This degree of improvement in cognitive
performance in healthy mice dosed just prior to testing and after learning has not been demonstrated with any cannabinoid compound previously.
Because
MIRA-55 is an anxiolytic, we decided to test whether it could impair cognitive function. We therefore sought to determine if MIRA-55
could impair attention-a different aspect of cognition than memory, recall and associative learning, and one that is affected negatively
by sedating compounds (e.g. THC, Cannabis, benzodiazepine, etc.) and positively by stimulants (e.g. caffeine, nicotine, amphetamine)
In order to assess whether MIRA-55 affected attention as compared to THC required a different testing model-Psychomotor Vigilance Test
(PVT). The rat Psychomotor Vigilance Test (rPVT) is a widely used method to measure sustained attention in rodents. In the rPVT model,
rats are trained to respond to a visual stimulus by pressing a lever, with shorter reaction times indicative of better attentional performance.
Mice with longer reaction times or higher variability in response times may be considered to have attention deficits or altered vigilance.
Data is shown as percentage accuracy at pressing the lever within the allowed reaction time vs dose of drug used. We have found that
at doses of THC that impair attention, MIRA-55 had no negative effects on attention (i.e. their accuracy at pressing a lever at the right
amount of time after receiving a trained cue was not impaired at all).
Status Planned Activity
Drug Substance Preparation Analytical Development and qualification
NonGMP Production Refinement and optimization
GLP/GMP Production Refinement
Testing Acute toxicity study mice
Genotoxicity studies
MTD/7D DRF Dog
MTD/7D DRF Rat
Dog 28-day Toxicology
Rat 28-day Toxicology
Cardiovascular Study Dog (Telemetry)
Respiratory Study Rat
hERG (Manual Patch-Clamp)
Neurobehavioral Evaluation Rats
Neurobehavioral Evaluation Mice
We
plan to conduct further neurobehavioral evaluations of orally administered MIRA-55 in rats and mice, along with a respiratory evaluation
of orally administered MIRA-55 in rats. Additionally, in vitro testing is planned to assess the effects of MIRA-55 on hERG (the human
Ether-à-go-go-Related Gene) channel currents, an early FDA-required assay that helps identify potential cardiac abnormalities
before advancing to human dose studies.
hERG
encodes a potassium ion channel that plays a critical role in the electrical activity of the heart. The hERG channel mediates the repolarizing
current in the cardiac action potential, ensuring proper heart rhythm. If the function of this channel is inhibited—whether by
drug interaction or rare genetic mutations—it can lead to long QT syndrome, a potentially life-threatening condition.
In
addition to these studies, we are planning a 28-day toxicology analysis in dogs and rats as part of the broader safety evaluation of
MIRA-55.
We
have also initiated analytical development and manufacturing for MIRA-55. Our suppliers are working toward scaling production under GLP/cGMP
conditions, building upon previous non-GMP batches used in initial testing. We are collaborating closely with our suppliers to generate
sufficient cGMP-grade MIRA-55 materials for planned preclinical toxicity programs, expanded animal testing, and potential human trials,
which will be conducted pending regulatory approval.
Regulation
The
FDA and comparable regulatory authorities in state and local jurisdictions impose substantial and burdensome requirements upon companies
involved in the clinical development, manufacture, marketing, and distribution of drugs. These agencies and other federal, state, and
local entities regulate, among other things, the research and development, testing, manufacture, quality control, safety, effectiveness,