Pipeline

Our Pipeline consists of drugs for chronic neurodegeneration – Alzheimer’s disease (AD), its orphan indication Alzheimer’s disease and dementia in Down syndrome (AD-DS) and Parkinson’s disease (PD). Additionally, we have a compound to treat acute neurodegeneration – traumatic brain injury (TBI) and stroke – and a third compound for advanced AD.

ANVS-401 is our lead compound. It is being developed for AD-DS, AD and PD, because in preclinical studies it normalized axonal transport in these diseases by inhibiting the neurotoxic proteins that kill nerve cells. It was shown in human and animal studies to lower APP/Aβ, tau/phospho-tau and α-synuclein, the neurotoxic proteins that impair axonal transport and lead to inflammation and cell death. The compound was tested in three Phase 1 clinical studies that show it to be well tolerated [Investigator Brochure 2015]. This safety data is applicable to the clinical development of ANVS-401 for AD-DS, AD, PD and other chronic neurodegenerative disorders.

ANVS-401 for AD. In the third Phase 1 clinical study ANVS-401 was tested in mildly cognitive impaired patients and was shown to normalize levels of APP, tau and αSYN in the cerebrospinal fluid (CSF). It also lowers and normalizes inflammatory factors in these patients [Maccecchini, 2012]. We have preclinical data showing that ANVS-401 lowered levels of APP and increased levels of neurotrophic factors in AD transgenic (tg) mice [Lijia, 2013]. Additionally, in a different study, ANVS-401 lowered APP and all its fragments and restored memory, learning and brain function in AD tg mice [Teich, 2018]. We have also conducted a study in tau tg mice showing that ANVS-401 lowers tau and phospho-tau.

ANVS-401 for AD-DS. We intend to initially focus on the orphan indication AD-DS. In DS the APP gene is triplicated leading to early onset AD with the same pathology and dementia as sporadic AD. This will allow us to obtain human data for AD in an orphan subpopulation much faster than in the regular AD population. Therefore, we plan to conduct our Phase 3 pivotal study in AD-DS rather than in AD. Studies in trisomic DS animals and nerve cells show that ANVS-401 fully restored and improved axonal transport, lowered APP, tau and phospho-tau and increased levels of BDNF (a neurotrophic factor), proving that normal axonal transport restored memory and learning and returned the brain to homeostasis  [Mobley, manuscript in preparation].

ANVS-401 for PD. We have tested ANVS-401 in PD tg mice that exhibit the pathology of PD. In these mice, treatment with ANVS-401 lowers aSYN in the gut and in the brain and fully normalizes colonic motility [Kuo, 2019].

ANVS-405.  For acute indications, we are developing ANVS-405, focused on protecting the brain after TBI and/or stroke. In a study, TBI rats that were treated with ANVS-405 after the insult exhibited normal memory and learning, normalized inflammation and re-established homeostasis in the brain [Chesselet, manuscript submitted for publication].

ANVS-301. Our next compound is ANVS-301. It is expected to increase cognitive capability in later stages of AD and dementia. In very old rats, ANVS-301 fully restored memory and learning and made the old rats cognitively equivalent to young rats. ANVS-301 is in a Phase 1 clinical trial that is being conducted and financed by the National Institutes of Health (NIH).

Clinical Trials

Safety

ANVS-401 is orally available, well behaved and well qualified as a candidate to treat neurodegeneration (Alzheimer’s disease – AD, Alzheimer’s disease in Down syndrome -AD-DS and Parkinson’s disease – PD). Three clinical studies have been conducted with ANVS-401. The first was a single ascending dose (SAD) study in 60 healthy volunteers. The second was a multiple ascending dose (MAD) study in 48 healthy volunteers. The third was a proof of concept (POC) study in five mildly cognitive impaired (MCI) patients.

Safety was evaluated in all three studies and ANVS-401’s pattern of adverse events (AEs) was similar to that seen in typical studies in healthy normal volunteers, with an overall incidence rate of 33.3% among placebo-treated subjects and 35% for all ANVS-401 treatment groups combined. In the single ascending dose study, the 160 mg/day group – the highest dose group in the study – had a treatment-related AE rate of 31.7%. In the multiple ascending dose and in the POC study there were no dose-related AEs. Most AEs were of short duration, mild or moderate in severity, and resolved without medical intervention. These data are reported to the FDA in SN 0016 and SN 0018.

Adverse events seen in the first human SAD study conducted with ANVS-401

Adverse events seen in the second and third human MAD and POC studies conducted with ANVS-401

Proof-of-Concept Study

In the human POC study, five patients with MCI were treated for 10 days with ANVS-401 with the safe dose of 4x60 mg (240 mg/day). Cerebrospinal fluid (CSF) and plasma were drawn over 12 hours on day 0 and day 11. Levels of ANVS-401 and metabolites were measured in plasma and CSF over the 12 hours. ANVS-401 pharmacokinetics (PK) in plasma corresponded to what the Company had seen in the previous clinical safety studies: T1/2 = 5 hrs. In CSF, however, ANVS-401 showed a much longer half-life of T1/2 > 12 hours [Maccecchini, 2012].

Annovis conducted an identical experiment in rats, where it is possible to measure the PK of ANVS-401 in plasma, CSF and the brain. Annovis extrapolated human plasma/CSF and rat plasma/CSF/brain levels to calculate human brain levels of ANVS-401 and determined they were eight times higher than plasma levels. This is consistent with the data the Company has in mice, where in a number of studies, ANVS-401 brain levels were found to be 6-10 times higher than plasma levels.

ANVS-401’s extended presence in the brain is matched by an extended effect, reducing levels of αSYN, APP and tau for the whole 12-hour period tested. The extended effect of ANVS-401 in human brains was consistent with the preclinical data in rodent brains.

The persistence of ANVS-401 in the CSF and brain and the extended pharmacodynamic effect observed make ANVS-401 a good candidate for once a day dosing. Extrapolated brain levels of ANVS-401 at 60 mg QID are far in excess of levels required to down-regulate APP and αSYN. The regulation of translation across the neurotoxic proteins is similarly responsive to ANVS-401, suggesting similar dosing in AD-DS, AD, PD and Huntington’s disease (HD).  Annovis further compared levels of ANVS-401 in the brains of mice that had shown full reversal of their AD or PD behavior and calculated that the optimum brain levels to achieve full efficacy are between 150 and 500 nM. Using three different extrapolation/comparison calculations the Company deduced that a daily dose of 10-50 mg should achieve desired brain levels and efficacy.

CSF Biomarkers Significantly Decrease After 10 Days of Oral ANVS-401 in MCI Patients

Human Biomarker

CSF % of Baseline

p-Value

sAPP α

-59.9%

0.0006

sAPP β

-57.7%

0.0001

Aβ42

-51.4%

0.053

Tau

-46.2%

0.002

p-Tau

-61.0%

0.0005

αSYN

-41.2%

0.091

MCI patients treated for 10 days and extracted CSF for 12 hours before and after the last administration of ANVS-401 show about 50% lower levels of aggregating toxic proteins; the levels measured have dropped to levels found in healthy normal volunteers.

Table: Inflammatory Markers after 10 days on ANVS-401

Human Inflammatory Protein

CSF % of Baseline

p-Value

Complement C3

-86.9%

0.0007

MCP-1

-87.5%

0.0007

YKL40

-72.7%

0.0113

sCD14

-26.1%

0.1159

Factor FH

23.7%

0.4988

MCI patients also show high levels of inflammatory factors and microglia activation factors in their CSF. ANVS-401 significantly lowered the levels of a number of inflammation factors in their CSF.

ANVS-401 normalizes the levels of toxic aggregating proteins to Levels Found in healthy Volunteers

Neurotoxic proteins measured in the CSF of MCI patients in the POC study described above. ANVS-401 given for 10 days does not just lower APP and Tau but normalizes it back to the levels seen in healthy volunteers.

Ongoing Annovis Phase 2a Study in AD Patients

We are presently conducting a study in early AD patients. We are treating patients for 4 weeks with ANVS-401 and measure target and pathway engagement in the spinal fluid.

This means that we are measuring levels of neurotoxic proteins, neurotransmitters, neurotrophic factors, inflammation and nerve cell death as well of cognitive improvement.

Planned Annovis Phase 2a Study in PD Patients

We are planning to start an equivalent Phase 2a study in early PD patients, in which we will also be measuring levels of neurotoxic proteins, neurotransmitters, neurotrophic factors, inflammation and nerve cell death as well of cognitive improvement.

Both studies:

  • Are multi-center, randomized, double-blind, placebo-controlled trials
  • Enroll, or will enroll, subjects diagnosed with early AD or early PD, respectively
  • Will show target and pathway engagement, proving that engaging the target stops the toxic cascade
  • Will potentially show efficacy
  • Will provide optimal information to proceed to pivotal studies, in AD-DS and PD, respectively, with large scale cognitive endpoints

Ongoing AD Study

Planned PD Study

Safety and Tolerability, PK Safety and Tolerability, PK

Primary Endpoints:

  • Effects on Aβ40 using SILK
  • Aβ38/40/42

Primary Endpoints:

  • L-Dopa, DaTscan
  • aSYN

Secondary Endpoints:

  • sAPPα, sAPPβ, p-tau, aSYN
  • AChE and neurotransmitters
  • Neurotrophic factors
  • Inflammatory markers
  • Other proteins

Secondary Endpoints:

  • sAPPα, sAPPβ, p-tau, aSYN
  • AChE and neurotransmitters
  • Nuerotrophic factors
  • Inflammatory markers
  • Other proteins

Cognitive Assessment:

  • ADAS-Cog 12, MMSE, NPI

Cognitive Assessment:

  • GDS short; HVLT-R; RBANS; TMT

The successful outcome of both studies will validate ANVS-401s inhibition of the toxic cascade that leads to neurodegeneration in AD and PD; it will give us some indications of efficacy; it will de-risk ANVS-401 and allow the company to move into two Phase 3 studies, one in AD-DS, an orphan indication of AD, and one in PD.

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