Treatment of viral infections, diseases, or disorders using selective S1R agonists

Abstract

The present invention provides a method for treating, reducing the incidence of, suppressing, or inhibiting a viral infection, disease, disorder, or symptom thereof in a subject in need thereof, comprising administering to the subject a selective S1R agonist. In another embodiment, the viral disease is COVID-19 and the selective S1R agonist is pridopidine or a pharmaceutically acceptable salt thereof.

Description

[Technical field]

[0001] The present invention provides a method for treating, reducing the incidence, suppressing or inhibiting a viral infection, disease, disorder, or symptoms thereof in a subject in need thereof, comprising administering to the subject a selective S1R agonist. In another embodiment, the viral disease is COVID-19 and the selective S1R agonist is pridopidine or a pharma- ceutically acceptable salt thereof. [Background technology]

[0002] Coronavirus disease 2019, or COVID-19, has rapidly emerged as a global pandemic. As of the end of April 2021, there are nearly 150 million confirmed cases in over 100 countries, with over 3 million deaths. Only a few therapeutics have received emergency approval, and vaccinations have only recently become available. Coronaviruses have single-stranded RNA genomes and encode many similar proteins. Protein and RNA replication machinery are classical targets for antiviral drugs in development over the past few years. However, the coronavirus life cycle depends on several host cell-encoded cellular pathways (Nabirotchkin et al. 2020). Among these pathways are the ER stress-induced unfolded protein response (UPR), autophagy, and mitochondrial function.

[0003] A recent bioinformatics / proteomics analysis (Gordon et al. 2020) identified SARS-CoV-2 proteins that interact with human Sigma-1 and Sigma-2 receptors (S1R / S2R) and contribute to disease pathogenesis. This highlights sigma receptors as potential drug targets for treating COVID-19. Among approximately 20 virus-encoded proteins, Nsp6 and Orf9c were identified as directly interacting with Sigma receptors.

[0004] S1R is an ER chaperone protein located in the mitochondria-associated membrane (MAM) that plays a key role in ER-mitochondria interaction. S1R regulates ER stress, mitochondrial function, calcium signaling, autophagy, and cell homeostasis (Weng, Tsai, and Su 2017; Delprat et al. 2020). Deletion of S1R enhances ER and oxidative stress, whereas overexpression and activation of S1R by different agonists restores cell homeostasis and improves survival. Activation of S1R has been shown to reduce ER stress, restore mitochondrial function, and enhance autophagy (Tesei et al. 2018; Maurice et al. 1994; Christ et al. 2019).

[0005] S2R is an intracellular chaperone protein that was recently cloned and identified as TMEM97 (Alon et al. 2017). Although S1R and S2R are not genetically related, they share similar pharmacological profiles, and some S1R ligands exhibit high affinity for S2R, including haloperidol and DTG (Longhitano et al. 2017; Tesei et al. 2018; Katnik et al. 2006). Some S2R ligands have been shown to induce apoptosis, making them attractive anticancer drugs (Tesei et al. 2019).

[0006] Several Sigma ligands have recently been reported to exhibit antiviral activity (Gordon et al. 2020). Sigma ligands that showed antiviral activity (measured by viral titer assay) included hydroxychloroquine, clemastine, and haloperidol. All of these compounds exhibit high affinity not only for S1R but also for S2R, and therefore all are nonselective compounds. The Ki for hydroxychloroquine is 200 nM and 800 nM for S1R and S2R, respectively, the Ki for clemastine is 10 nM for S1R and 20 nM for S2R, and haloperidol has a Ki for S1R of 4 nM and a Ki for S2R of 54 nM.

[0007] Gordon, in his paper, presents antiviral activity with hydroxychloroquine, clemastine, and haloperidol (Gordon et al. 2020). Antiviral activity of hydroxychloroquine was shown at approximately 2 uM, which was associated with a decrease in cell viability of approximately 30% (Gordon et al. 2020). The effective clemastine dose for antiviral activity was 10 uM, which was also associated with approximately 40% cell death. Similarly, antiviral effect of haloperidol was shown at 100 uM, with approximately 30% cell death (Gordon et al. 2020).

[0008] Pridopidine (4-[3-(methylsulfonyl)phenyl]-1-propyl-piperidine) is a highly selective S1R ligand with a Ki of 0.57 nM and a Ki of 5450 nM for S2R (Johnston et al. 2019). Thus, pridopidine has a 95-fold higher affinity for S1R compared to S2R, making it the most selective S1R ligand. Summary of the Invention [Means for solving the problem]

[0009] In a first aspect, the present invention provides a method for treating, reducing the incidence, suppressing or inhibiting a viral infection, disease, disorder, or symptoms thereof in a subject in need thereof, comprising administering to the subject a composition comprising a selective S1R agonist.

[0010] In a further aspect, the present invention provides a method for reducing endoplasmic reticulum stress (ER stress) resulting from a viral infection, disease, or disorder in a subject, the method comprising administering to the subject a composition comprising a selective S1R agonist.

[0011] In a further aspect, the invention provides a method for treating, reducing the incidence, suppressing or inhibiting a human coronavirus or a symptom thereof or a mutation thereof in a subject in need thereof, comprising administering to the subject a composition comprising a selective S1R agonist.

[0012] In a further aspect, the present invention provides a method for treating, reducing the incidence, suppressing or inhibiting a viral infection, disease, disorder, or symptoms thereof in a subject in need thereof, comprising administering a composition comprising pridopidine, a pharma- ceutically acceptable salt thereof, a deuterated analog thereof, or a combination of pridopidine and at least one of its analog compounds 1-7 or a salt thereof.

[0013] [ka]

[0014] In some embodiments, the viral infection, disease, or disorder comprises a human coronavirus, Severe Acute Respiratory Syndrome (SARS), Middle East Respiratory Syndrome (MERS) coronavirus, SARS coronavirus 2 (SARS-CoV-2), or a mutation therein, hi other embodiments, the disease is COVID-19.

[0015] In some embodiments, the methods of the present invention use a selective S1R agonist. In another embodiment, the selective S1R agonist is pridopidine, a pharma- ceutically acceptable salt thereof, a deuterated analog thereof, or a combination of pridopidine and at least one of analogs 1-7 or a salt thereof.

[0016] In some embodiments, the pridopidine is in its neutral / base form. In some embodiments, the pridopidine is in a pharma- ceutically acceptable salt form. In some further embodiments, the pridopidine is pridopidine hydrochloride.

[0017] In some embodiments, the composition comprising the selective S1R agonist is administered orally. In some embodiments, the composition comprising pridopidine, a pharma- ceutically acceptable salt thereof, a deuterated analog thereof, or a combination of pridopidine and at least one of analogs 1-7 or a salt thereof is administered orally.

[0018] In another embodiment, the composition is administered by systemic administration. In another embodiment, the composition is administered by oral administration. In another embodiment, the composition is formulated as an oral liquid, solid, semi-solid dosage form, injectable solution, dermal / transdermal dosage form, ophthalmic dosage form, inhalable composition. In another embodiment, the composition is formulated as an inhalable powder, injectable solution, liquid, gel, solid, capsule, eye drop, or tablet.

[0019] In some embodiments, the composition is administered periodically (i.e., the pridopidine is administered periodically at predetermined time intervals, such as daily, hourly, weekly, monthly periods, each optionally also defining the dose administered and the number of doses per period). In further embodiments, the composition is administered once a day, twice a day, or three times a day. In further embodiments, the composition is administered less frequently than once a day. In some embodiments, the composition is administered in one dose, two doses, or three doses per day. [Brief description of the drawings]

[0020] The subject matter which is regarded as the invention is particularly pointed out and distinctly claimed in the concluding portion of this specification, however the invention, both as to organization and method of operation, together with its objects, features, and advantages, may best be understood by reference to the following detailed description taken in conjunction with the accompanying drawings.

[0021] [Figure 1] Representative images are shown showing that mutant Htt (mHtt) results in pridopidine-induced ER stress (H2a-GFP) quantified by aggregation of H2a-GFP. Early ER stress is quantified with H2a-GFP, a protein indicator that accumulates to form aggregates in response to ER stress. H2a-GFP was transiently co-expressed with wild-type (WT, Htt20Q)-mCherry or mutant (Htt96Q)-mCherry (exon 1) in STHdhQ7 / 7 cells. Starting 4 hours after transfection, cells were treated with no or increasing concentrations of pridopidine (from 0.03 μM to 3 μM) and imaged by confocal microscopy 24 hours after transfection. Images of individual cells (approximately 150 cells per experiment) containing Htt96Q-mCherry aggregates or Htt20Q-mCherry were quantified relative to untreated cells with or without aggregates. [Diagram 2] Quantification of ER stress in cells with mHTT (left) or wild-type HTT (right) is shown. Pridopidine significantly reduces early mutant Htt-induced ER stress in a dose-dependent manner, as measured by the aggregation of H2a-GFP. For comparison purposes, 100% represents the relative intensity of H2a-GFP in untreated cells exhibiting mHtt-mCherry aggregates, and 0% is the relative intensity of H2a-GFP in untreated cells without mHtt-mcherry aggregates. Graphs are the mean ± SE of three experiments. Asterisks indicate P values ​​of <0.05 (*) and <0.01 (**) compared to untreated. Wild-type HTT (right) does not induce ER stress and no effect is observed. [Diagram 3] Pridopidine reduces mutant Htt-induced eIF2α phosphorylation (ER stress marker) at 8 h. HEK293 cells were transfected with myc-Htt96Q (mutant HTT, squares) or myc-Htt20Q (wild-type HTT, circles) and then treated with increasing concentrations (from 0.03 μM to 3 μM) of pridopidine for 8 h. The ratio of eIF2a-P to total eIF2a was quantified by immunoblot. Pridopidine reduces the levels of eIF2a-P in a dose-dependent manner, with a significant effect at the 3 μM dose (p<0.01). [Figure 4] Pridopidine reduces mutant Htt-induced eIF2α phosphorylation (ER stress marker) at 24 h. HEK293 cells were transfected with myc-Htt96Q (mutant HTT, squares) or myc-Htt20Q (wild-type HTT, circles) and then treated with increasing concentrations (from 0.03 μM to 3 μM) of pridopidine for 24 h. The ratio of eIF2a-P to total eIF2a was quantified by immunoblot. Pridopidine reduces the levels of eIF2a-P in a dose-dependent manner, with significant effects at 0.3 μM and 3 μM doses (p<0.05). [Diagram 5] Pridopidine reduces eIF2α phosphorylation (an ER stress marker) in cells in which ER stress is induced by thapsigargin. Thapsigargin is a potent inducer of ER stress. HEK293 cells transfected with myc-Htt20Q (wild-type HTT) are treated with or without 2 μg / ml thapsigargin, which causes a 70% increase in phosphorylated eIF2α (eIF2α-p). Pridopidine treatment reduces p-eIF2α levels (3 μM). [Figure 6]Pridopidine reduces eIF2α-phosphorylation (ER stress marker) in an S1R-dependent mechanism. HEK293 cells were transfected with guide RNAs (gRNAs) targeting human S1R or control gRNA. Htt96Q (mHTT) was transiently expressed in HEK293 cells. Cells were treated with 0.3 μM and 3 μM pridopidine for 8 h. The ratio of eIF2α-P to total eIF2α was measured and quantified by immunoblotting. Pridopidine treatment reduces p-eIF2α levels at concentrations of 0.3 μM and 3 μM in cells expressing S1R (S1R+ / +) (p<0.01). However, this effect is ineffective in the absence of S1R (S1R- / - cells), indicating an ER stress-reducing effect of pridopidine mediated by S1R. Data are the mean ± SD of three experiments. [Figure 7A] Pridopidine reduces the protein levels of the unfolded protein response pathway (UPR) markers ATF4 (Figure 7A), CHOP (Figure 7B) and GADD34 (Figure 7C). Cells expressing myc-Htt96Q (mutant Htt) or myc-Htt20Q (wild-type Htt) were treated with 3 uM pridopidine. Immunoblots were reacted with anti-ATF4 (Figure 7A), anti-CHOP (Figure 7B) and anti-GADD34 (Figure 7C), quantified and normalized with anti-actin or anti-tubulin as loading controls. Pridopidine (3 μM) reduces ATF4 levels by approximately 4.5-fold (p<0.05), CHOP by approximately 2-fold and GADD-34 by approximately 2.5-fold (p<0.01). [Figure 7B]Pridopidine reduces the protein levels of the unfolded protein response pathway (UPR) markers ATF4 (Figure 7A), CHOP (Figure 7B) and GADD34 (Figure 7C). Cells expressing myc-Htt96Q (mutant Htt) or myc-Htt20Q (wild-type Htt) were treated with 3 uM pridopidine. Immunoblots were reacted with anti-ATF4 (Figure 7A), anti-CHOP (Figure 7B) and anti-GADD34 (Figure 7C), quantified and normalized with anti-actin or anti-tubulin as loading controls. Pridopidine (3 μM) reduces ATF4 levels by approximately 4.5-fold (p<0.05), CHOP by approximately 2-fold and GADD-34 by approximately 2.5-fold (p<0.01). [Figure 7C] Pridopidine reduces the protein levels of the unfolded protein response pathway (UPR) markers ATF4 (Figure 7A), CHOP (Figure 7B) and GADD34 (Figure 7C). Cells expressing myc-Htt96Q (mutant Htt) or myc-Htt20Q (wild-type Htt) were treated with 3 uM pridopidine. Immunoblots were reacted with anti-ATF4 (Figure 7A), anti-CHOP (Figure 7B) and anti-GADD34 (Figure 7C), quantified and normalized with anti-actin or anti-tubulin as loading controls. Pridopidine (3 μM) reduces ATF4 levels by approximately 4.5-fold (p<0.05), CHOP by approximately 2-fold and GADD-34 by approximately 2.5-fold (p<0.01). [Figure 8] Pridopidine reduces the protein levels of the UPR marker ATF6. Cells expressing myc-Htt96Q (mutant Htt) were treated with pridopidine at 0.03 μM and 3 μM. Immunoblots were reacted with anti-ATF6, quantified, and normalized with anti-tubulin as a loading control. Both concentrations of pridopidine reduce ATF6 by approximately 10% and approximately 30% at concentrations of 0.03 μM and 3 μM, respectively. [Figure 9]Pridopidine reduces the mRNA levels of the UPR marker XBP1s. Cells expressing myc-Htt96Q (mutant Htt) or myc-Htt20Q (wild-type Htt) were treated with pridopidine at 0.03 μM and 3 μM. RNA levels of XBP1s were determined by quantitative PCR (polymerase chain reaction). Xbp1s levels are increased by 70% in mutant Htt-transfected cells. Pridopidine treatment shows a significant 25% reduction in XBP1s levels at 3 μM concentration (p<0.01). [Figure 10] Pridopidine reduces mitochondrial reactive oxygen species (ROS) in YAC128 HD striatal neurons. YAC128 is a mouse model of Huntington's disease (HD), and wild-type or YAC128-derived striatal neurons were treated with or without 1 μM pridopidine and incubated with MitoPY1 fluorescent probe. Mitochondrial H2O2 release and oxidative stress were induced using the mitochondrial respiration inhibitor antimycin A (Ant A, 2 μM). Mitochondrial H2O2 was recorded with spinning disk confocal before and after administration of Ant A (n=4, considering about 20 cells / condition). Ant A increased H2O2 in untreated cells by about 2-fold. Pridopidine treatment inhibits H2O2 release and oxidative stress. Scale bar=30 μM. Two-way ANOVA reveals the rescue effect of pridopidine treatment on mitochondrial ROS production [F(1,389)=15.24; p<0.0001]. [Figure 11]Pridopidine increases mitochondrial membrane potential (MMP) in cortical HD neurons following oxidative stress. Wild-type and YAC128 (HD) derived striatal neurons were treated with pridopidine for 24 h and TMRM (tetramethylrhodamine methyl ester) was used to assess changes in MMP after depolarization with oligomycin + FCCP (carbonyl cyanide-4-phenylhydrazone) (n = 7–10). HD neurons show reduced MMP. Pridopidine treatment enhances MMP in wild-type neurons and significantly restores MMP impairment in HD neurons (2-way ANOVA analysis [F(2,107) = 3.257; p = 0.0423]). [Figure 12] The effect of pridopidine on mitochondrial membrane potential (MMP) is mediated by S1R. Genetic knockdown (KD) of S1R in lymphoblasts from HD patients (S1R KD) showed approximately 83% reduction in S1R protein levels. Cells were treated with pridopidine 5 μM and then challenged with 0.1 mM H2O2. MMP was quantified by TMRE (tetramethylrhodamine ethyl ester) signal in control and S1R-KD HD lymphoblasts (5 μM, 24 h, n=4). H2O2 treatment significantly reduced MMP by 25% and 75% in both S1R+ / + and S1R-KD cells, respectively. Pridopidine treatment completely restored MMP in S1R+ / + cells, but not in S1R KD cells, indicating that the effect of pridopidine is mediated by S1R. ****p<0.0001 Kruskal-Wallis test followed by Dunn's multiple comparison test. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0022] In the following detailed description, numerous specific details are set forth in order to provide a thorough understanding of the present invention. However, it will be understood by those skilled in the art that the present invention may be practiced without these specific details. In other instances, well-known methods, procedures, and components have not been described in detail so as not to obscure the present invention.

[0023] In a first aspect, the present invention provides a method for treating, reducing the incidence, suppressing or inhibiting a viral infection, disease, disorder, or symptoms thereof in a subject in need thereof, comprising administering to the subject a composition comprising a selective S1R agonist.

[0024] In a further aspect, the present invention provides a method for treating, reducing the incidence, suppressing or inhibiting a viral infection, disease, disorder, or symptoms thereof in a subject in need thereof, comprising administering a composition comprising pridopidine, a pharma- ceutically acceptable salt thereof, a deuterated analog thereof, or a combination of pridopidine and at least one of its analog compounds 1-7 or a salt thereof.

[0025] [ka]

[0026] When referring to a "viral infection, disease, disorder, or any symptom thereof," it should be understood to encompass any type of condition that endangers the health of a subject in which a viral infection plays a direct or indirect role.

[0027] In some embodiments, the viral infection, disease, or disorder comprises a human coronavirus, Severe Acute Respiratory Syndrome (SARS), Middle East Respiratory Syndrome (MERS) coronavirus, SARS coronavirus 2, or a mutation therein. In other embodiments, the disease treated by administering a composition comprising a selective S1R agonist is COVID-19.

[0028] In one embodiment, the present invention provides a method of treating, reducing the incidence, suppressing or inhibiting a human coronavirus or a mutation thereof and / or a symptom thereof by administering a composition comprising a selective S1R agonist.

[0029] In one embodiment, the present invention provides a method of treating, reducing the incidence, suppressing or inhibiting Severe Acute Respiratory Syndrome (SARS) or variants thereof and / or symptoms thereof by administering a composition comprising a selective S1R agonist.

[0030] In one embodiment, the present invention provides a method of treating, reducing the incidence, suppressing or inhibiting Middle East Respiratory Syndrome (MERS) coronavirus or its mutations and / or symptoms thereof by administering a composition comprising a selective S1R agonist.

[0031] In one embodiment, the invention provides a method of treating, reducing the incidence, suppressing or inhibiting SARS coronavirus 2 (SARS-CoV-2) or a mutation thereof and / or symptoms thereof by administering a composition comprising a selective S1R agonist.

[0032] In one embodiment, the present invention provides a method of treating, reducing the incidence, suppressing or inhibiting COVID-19 or its mutations and / or symptoms thereof by administering a composition comprising a selective S1R agonist.

[0033] In one embodiment, the present invention provides a method of reducing ER stress resulting from a viral infection, disease, or disorder in a subject. In another embodiment, the viral infection, disease, or disorder comprises a human coronavirus, SARS, MERS coronavirus, SARS coronavirus 2, or a mutation and / or symptom thereof. In another embodiment, the disease is COVID-19 by administering a composition comprising a selective S1R agonist.

[0034] In some embodiments, the present invention provides methods of treating, reducing the incidence, suppressing or inhibiting Middle East Respiratory Syndrome (MERS) coronavirus or its mutations and / or symptoms thereof by administering a composition comprising a selective S1R agonist. In other embodiments, the symptoms include renal failure, fever, fatigue, dry cough, body aches, nasal congestion, runny nose, sore throat, diarrhea, or a combination thereof.

[0035] The most common symptoms of COVID-19 are fever, dry cough, and fatigue. Some patients may also have body aches, nasal congestion, sore throat, or diarrhea. These symptoms are usually mild and develop gradually. Some people only have very mild symptoms when infected. Most people (about 80%) recover from the disease without needing hospital treatment. About one in five people infected with COVID-19 become seriously ill and develop difficulty breathing. Older people and people with underlying conditions such as high blood pressure, heart and lung problems, diabetes, or cancer are at higher risk of developing severe disease. However, anyone can become infected with COVID-19 and become seriously ill. Even people with very mild symptoms of COVID-19 can transmit the virus. People of any age who experience fever, cough, or difficulty breathing should seek medical attention.

[0036] In some embodiments, the present invention provides methods of treating, reducing the incidence, suppressing or inhibiting COVID-19 by administering a composition comprising a selective S1R agonist. In other embodiments, symptoms include renal failure, fever, fatigue, dry cough, body aches, nasal congestion, runny nose, sore throat, diarrhea, or a combination thereof.

[0037] In some embodiments, the methods of the present invention use a composition comprising a selective S1R agonist. In other embodiments, the S1R agonist is pridopidine, a pharma- ceutically acceptable salt thereof, a deuterated analog thereof, or a combination of pridopidine and at least one of analogs 1-7 or a salt thereof.

[0038] SAR-CoV-2 infection induces the unfolded protein response (UPR) / autophagy pathway, which is essential for the coronavirus life cycle.

[0039] In response to viral invasion, host cells attempt to restore endoplasmic reticulum (ER) homeostasis by activating the unfolded protein response (UPR) and shutting down global protein translation.

[0040] Induction of the UPR by coronavirus invasion constitutes a major aspect of virus-host cell interactions. ER stress and UPR activation contribute significantly to viral replication and viral pathogenesis during coronavirus infection (Fung and Liu 2014).

[0041] Viruses manipulate the UPR to complete their life cycle and enhance their propagation (Cava, Bertoli, and Castiglioni 2020), therefore, alleviating ER stress is an attractive candidate target for antiviral therapy.

[0042] Previously identified coronaviruses such as SARS-CoV and MERS-CoV have been shown to induce ER stress, utilize the ER for viral replication, and disrupt apoptotic pathways to ensure continued viral replication (S. Li et al. 2020, DeDiego et al. 2011). SARS-CoV induction of the UPR has been suggested to selectively regulate its function to enhance its replication while avoiding apoptosis (Chan et al. 2006).

[0043] Viral infection-induced ER stress leads to phosphorylation of eIF2α, inhibiting global translation in cells but increasing the translation of UPR-related genes such as ATF4 and CHOP (Bechill et al. 2008; K. Liao et al. 2016).

[0044] In some embodiments, the selective S1R agonist reduces ER stress in a patient suffering from a viral infection, disease, or disorder. In other embodiments, the selective S1R agonist is pridopidine, a pharma- ceutically acceptable salt thereof, a deuterated analog thereof, or a combination of pridopidine and at least one of analogs 1-7 or a salt thereof.

[0045] The UPR pathway, together with the autophagy pathway, is essential for viral infection, regulation of proteostasis, innate immunity, and clearance of viral particles. For example, the coronavirus MERS-CoV blocks the autophagy pathway (Gassen et al. 2019).

[0046] In some embodiments, the selective S1R agonist modulates ER stress and UPR pathways in patients suffering from a viral infection, disease, or disorder. In another embodiment, the selective S1R agonist reduces ER stress. In another embodiment, the selective S1R agonist is pridopidine, a pharma- ceutically acceptable salt thereof, a deuterated analog thereof, or a combination of pridopidine and at least one of analogs 1-7 or a salt thereof.

[0047] In some embodiments, the selective S1R agonist regulates the autophagy pathway in patients suffering from a viral infection, disease, or disorder. In other embodiments, the selective S1R agonist is pridopidine, a pharma- ceutically acceptable salt thereof, a deuterated analog thereof, or a combination of pridopidine and at least one of analogs 1-7 or a salt thereof.

[0048] Oxidative stress and mitochondrial dysfunction in COVID-19

[0049] Mitochondria play a key role in controlling ROS levels, with dysfunctional mitochondria producing high levels of uncontrolled toxic ROS (excessive ROS levels cause extensive damage to cells and oxidize proteins). Specifically, it has been shown that SARS-CoV2 infection enhances ROS levels, mainly due to the activity of the immune system (Wang, Zhang, and Bai 2020; Starkov 2008). The ORF8 protein of SARS-CoV localizes to mitochondria, where it causes increased ROS production, indicating a role in regulating ROS production and mitochondrial function (Chen et al. 2007). In light of this, treatments that can reduce ROS are potential targets for COVID-19.

[0050] In some embodiments, the selective S1R agonist reduces ROS in a patient suffering from a viral infection, disease, or disorder. In other embodiments, the selective S1R agonist is pridopidine, a pharma- ceutically acceptable salt thereof, a deuterated analog thereof, or a combination of pridopidine and at least one of analogs 1-7 or a salt thereof.

[0051] Patients infected with SARS-CoV-2 have high plasma levels of pro-inflammatory cytokines, including IL-1β, IL-2, TNFα, MCP1, IL7, and GSCF (Harapan et al. 2020). Increased cytokine levels are a common complication of respiratory disease, and overproduction of pro-inflammatory cytokines, an early response, can lead to severe complications, including multiple organ failure and death (Ricardo J Jose and Manuel 2020). ICU patients have significantly higher levels of GSCF, MCP1, and TNFα, suggesting that cytokine storm may be an underlying cause of disease severity. Treatment with S1R agonists has been shown to reduce cytokine levels and suppress inflammatory responses (Zhao et al. 2014, Allahtavakoli and Jarrott 2011).

[0052] In some embodiments, the selective S1R agonist reduces cytokine plasma levels in a patient suffering from a viral infection, disease, or disorder. In other embodiments, the selective S1R agonist is pridopidine, a pharma- ceutically acceptable salt thereof, a deuterated analog thereof, or a combination of pridopidine and at least one of analogs 1-7 or a salt thereof.

[0053] Compositions for use in the methods of the invention

[0054] In some embodiments, the present invention provides a composition comprising a selective S1R agonist for use in the methods of the present invention. In some embodiments, the present invention provides a composition comprising pridopidine, a pharma- ceutically acceptable salt thereof, a deuterated analog thereof, or a combination of pridopidine and at least one of analogs 1-7 or a salt thereof for use in the methods of the present invention.

[0055] In some embodiments, the present invention provides compositions comprising a selective S1R agonist for use in treating, reducing the incidence, suppressing or inhibiting a viral infection, disease, disorder, or symptoms thereof in a subject in need thereof.

[0056] In some embodiments, the present invention provides a composition comprising pridopidine, a pharma- ceutically acceptable salt thereof, a deuterated analog thereof, or a combination of pridopidine and at least one of its analog compounds 1-7 or a salt thereof, for use in treating, reducing the incidence, suppressing or inhibiting a viral infection, disease, disorder, or a symptom thereof in a subject in need thereof.

[0057] In some embodiments, the present invention provides compositions comprising a selective S1R agonist for use in reducing endoplasmic reticulum stress (ER stress) resulting from a viral infection, disease, or disorder in a subject in need thereof.

[0058] In some embodiments, the present invention provides a composition comprising pridopidine, a pharma- ceutically acceptable salt thereof, a deuterated analog thereof, or a combination of pridopidine and at least one of its analog compounds 1-7 or a salt thereof, for use in reducing endoplasmic reticulum stress (ER stress) due to a viral infection, disease, or disorder in a subject in need thereof.

[0059] In some embodiments, the compositions for use in the methods of the present invention include pridopidine, or a pharma- ceutically acceptable salt thereof. In another embodiment, pridopidine is neutral / free base. In another embodiment, pridopidine is in its pharma- ceutically acceptable salt form.

[0060] In another embodiment, the pridopidine salt is pridopidine hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, isonicotinate, acetate, lactate, salicylate, citrate, D,L-tartrate, L-tartrate, D-tartrate, pantothenate, bitartrate, ascorbate, succinate, hemisuccinate, maleate, gentisinate, geminate, or tetrahydrofuran. In another embodiment, the pridopidine salt is pridopidine hydrochloride.

[0061] In some embodiments, a composition for use in the methods of the present invention comprises pridopidine in combination with at least one of its analogues 1-7 or a salt thereof.

[0062] [ka]

[0063] In other embodiments, pridopidine analogs 1-7 and methods for their preparation can be found in U.S. Pat. Nos. 10,130,621 and 10,406,145, the entire contents of each of which are incorporated herein by reference.

[0064] In some embodiments, the composition for use in the method of the invention comprises pridopidine or a pharma- ceutically acceptable salt thereof in combination with at least Compound 1 or a pharma- ceutically acceptable salt thereof. In other embodiments, the composition for use in the method of the invention comprises pridopidine or a pharma- ceutically acceptable salt thereof in combination with Compound 4 or a pharma- ceutically acceptable salt thereof. In other embodiments, the composition for use in the method of the invention comprises pridopidine in combination with Compound 1 or a pharma- ceutically acceptable salt thereof, and Compound 4 or a pharma- ceutically acceptable salt thereof. In other embodiments, the composition for use in the method of the invention comprises pridopidine or a pharma- ceutically acceptable salt thereof in combination with at least Compound 2 or a pharma- ceutically acceptable salt thereof. In other embodiments, the composition for use in the method of the invention comprises pridopidine or a pharma- ceutically acceptable salt thereof in combination with at least Compound 3 or a pharma- ceutically acceptable salt thereof. In other embodiments, the composition for use in the method of the invention comprises pridopidine or a pharma- ceutically acceptable salt thereof in combination with at least Compound 4 or a pharma- ceutically acceptable salt thereof. In another embodiment, a composition for use in the methods of the invention comprises pridopidine or a pharma- ceutically acceptable salt thereof in combination with at least compound 5, or a pharma- ceutically acceptable salt thereof. In another embodiment, a composition for use in the methods of the invention comprises pridopidine or a pharma- ceutically acceptable salt thereof in combination with at least compound 6, or a pharma- ceutically acceptable salt thereof. In another embodiment, a composition for use in the methods of the invention comprises pridopidine or a pharma- ceutically acceptable salt thereof in combination with at least compound 7, or a pharma- ceutically acceptable salt thereof.

[0065] In another embodiment, a composition for use in the method of the invention comprises pridopidine or a pharma- ceutically acceptable salt thereof in combination with at least one of compounds 1-7, wherein at least one of compounds 1-7 is present at 0.01% to 5% by weight of the composition. In another embodiment, a composition for use in the method of the invention comprises pridopidine or a pharma- ceutically acceptable salt thereof in combination with at least one of compounds 1-7, wherein at least one of compounds 1-7 is present at 0.01% to 1% by weight, 0.05 to 0.5% by weight, or 0.05% to 1% by weight of the composition. In another embodiment, a composition for use in the method of the invention comprises pridopidine or a pharma- ceutically acceptable salt thereof in combination with compound 1 or a pharma- ceutically acceptable salt thereof, wherein compound 1 is present at 0.01% to 5% by weight, 0.01% to 1% by weight, 0.05 to 0.5% by weight, or 0.05% to 1% by weight of the composition. In another embodiment, a composition for use in the methods of the invention comprises pridopidine or a pharma- ceutically acceptable salt thereof in combination with compound 4 or a pharma- ceutically acceptable salt thereof, wherein compound 4 is 0.01% to 5%, 0.01% to 1%, 0.05 to 0.5%, or 0.05% to 1% by weight of the composition.

[0066] In some embodiments, the present invention provides compositions comprising deuterated analogs of pridopidine for use in the methods of the present invention. "Deuterated analog" refers to a "deuterium-enriched" compound in which the abundance of deuterium at any relevant site of the compound in a given amount of the compound is greater than the abundance of deuterium naturally occurring at that site. The distribution of naturally occurring deuterium is approximately 0.0156%. Thus, in a "deuterium-enriched" compound, the abundance of deuterium at any of its relevant sites is greater than 0.0156%, and may range from greater than 0.0156% to 100%. Deuterium-enriched compounds can be obtained by exchanging hydrogen for deuterium or by synthesizing the compound with deuterium-enriched starting materials.

[0067] In other embodiments, examples of deuterated analogs of pridopidine and methods for their preparation can be found in U.S. Patent Application Publication Nos. 2013-0197031, 2016-0166559, and US2019-0015401, the entire contents of each of which are incorporated herein by reference.

[0068] In other embodiments, the deuterated analog of pridopidine is selected from:

[0069] [ka]

[0070] For the methods and uses disclosed herein, the route of administration may be, for example, oral. Routes of administration may also be classified according to whether the effect is local (e.g., in topical administration) or systemic (e.g., in enteral or parenteral administration). As used herein, "local administration" refers to the direct administration of a compound or composition to the location where its action is desired, and specifically excludes systemic administration. As used herein, "topical administration" of a compound or composition shall mean application of the compound or composition to a body surface, such as the skin or mucous membrane, such as the eye. As used herein, "ocular administration" refers to application of a compound or composition to the subject's eye, or the skin around the eye (periocular skin), or the mucous membrane around the eye, specifically the conjunctiva of the subject, i.e., local administration.

[0071] The amount of the pridopidine or selective S1R agonist and pharmaceutical compositions of the present invention may be administered orally, topically, systemically, locally, or ocularly.

[0072] In some embodiments, the compositions disclosed herein for use in the methods of the invention are administered by systemic administration. In other embodiments, the compositions are administered by oral administration. In other embodiments, the compositions are formulated as oral liquids, solids, semi-solid dosage forms, injectables, dermal / transdermal dosage forms, ophthalmic dosage forms, or as inhalable compositions. In other embodiments, the compositions are formulated as inhalable powders, injectables, liquids, gels, solids, capsules, eye drops, or as tablets.

[0073] In some embodiments, the compositions disclosed herein for use in the methods of the invention are administered once per day, twice per day, three times per day, or less than once per day.

[0074] In some embodiments, the compositions disclosed herein for use in the methods of the invention are administered in one dose, two doses, or three doses per day.

[0075] Examples of pridopidine derivatives are deuterium-enriched pridopidine and salts. Examples of deuterium-enriched pridopidine and salts and their preparation methods can be found in U.S. Patent Application Publication Nos. 2013-0197031, 2016-0166559 and 2016-0095847, the entire contents of each of which are incorporated herein by reference.

[0076] The present invention also includes any salt of pridopidine, including any pharmaceutically acceptable salt, where pridopidine has a net charge (either positive or negative) and at least one counter ion (with opposite negative or positive charge) is added to it to form the salt.As used herein, the phrase "pharmaceutically acceptable salt" refers to a salt of the compound of the present invention that is safe and effective for pharmaceutical use in mammals and has the desired biological activity.Pharmaceutically acceptable salts include salts of acid or base groups present in the compound of the present invention. Pharmaceutically acceptable acid addition salts include pridopidine hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, isonicotinate, acetate, lactate, salicylate, citrate, D,L-tartrate, L-tartrate, D-tartrate, pantothenate, bitartrate, ascorbate, succinate, hemisuccinate, maleate, gentisinate, gentisic acid, fumarate, and the like. The salts of pridopidine include, but are not limited to, salts of lutetate, gluconate, glucuronate, glycolate, saccharate, formate, besylate, benzoate, glutamate, malate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, oxalate, tosylate, naphthalene-2-sulfate, or pamoate (i.e., 1,1'-methylene-bis-(2-hydroxy-3-naphthoic acid)) salts. Certain compounds of the invention can form pharma- ceutically acceptable salts with various amino acids. Suitable base salts include, but are not limited to, aluminum, calcium, lithium, magnesium, potassium, sodium, zinc, and diethanolamine salts. For a review of pharma- ceutically acceptable salts, see BERGE ET AL., 66 J.PHARM.SCI.1-19 (1977), incorporated herein by reference. In another embodiment, the pridopidine salt of the invention is a hydrochloride salt.

[0077] Accordingly, the present invention also relates to pharmaceutical compositions comprising the agents of the present invention in a mixture with pharma- ceutically acceptable adjuvants, and optionally other therapeutic agents. The adjuvant must be "acceptable" in the sense of being compatible with the other ingredients of the composition and not deleterious to the receptors thereof.

[0078] Pharmaceutical compositions include those suitable for oral, rectal, nasal, topical (including transdermal, buccal and sublingual), vaginal or parenteral (including subcutaneous, intramuscular, intravenous and intradermal) administration or by implant. The compositions may be prepared by any method well known in the art of pharmacy.

[0079] Pharmaceutical compositions include those suitable for oral, rectal, nasal, topical (including transdermal, buccal and sublingual), vaginal or parenteral (including subcutaneous, intramuscular, intravenous and intradermal) administration or by implant. The compositions may be prepared by any method well known in the art of pharmacy.

[0080] Such methods include the step of incorporating the associative compounds used in the present invention or combinations thereof together with any auxiliary agents, also referred to as accessory ingredients, including conventional ones such as carriers, fillers, binders, diluents, disintegrants, lubricants, colorants, flavorants, antioxidants, and wetting agents.

[0081] Pharmaceutical compositions suitable for oral administration may be presented as discrete dosage units such as pills, tablets, dragees or capsules, or as powders or granules, or as a solution or suspension. The active ingredient may be presented as a bolus or paste. The composition may be further processed into a suppository or enema for rectal administration.

[0082] The invention further includes a pharmaceutical composition as described above in combination with packaging material containing instructions for use of the composition as described above.

[0083] For parenteral administration, suitable compositions include aqueous and non-aqueous sterile injections. The compositions may be presented in unit-dose or multi-dose containers, such as sealed vials and ampoules, and may be stored in a freeze-dried (lyophilized) state, requiring only the addition of a sterile liquid carrier, such as water, prior to use. For transdermal administration, for example, gels, patches, or sprays may be contemplated. Suitable compositions or formulations for pulmonary administration, such as nasal inhalation, include fine dusts or sprays that can be generated by pressurized metered-dose aerosols, nebulizers, or inhalers.

[0084] The exact dosage and administration regimen of the compositions will necessarily depend on the therapeutic or nutritional effect to be achieved and may vary with the formula, route of administration, and the age and condition of the individual subject to which the composition is administered.

[0085] The term "treatment" as used herein refers to administering a therapeutic amount of the composition of the present invention effective to ameliorate an undesirable disease, disorder (including symptoms associated with the disease or disorder), prevent the appearance of such disease, disorder (including symptoms associated with the disease or disorder) before they occur, slow the progression of the disease, slow the worsening of symptoms, enhance the onset of remission, slow irreversible damage caused by the progressive chronic phase of the disease, delay the onset of the progressive phase, reduce the severity of the disease or cure the disease, improve survival or more rapid recovery, or prevent the onset of the disease, or a combination of two or more of the above. An "effective amount" for purposes disclosed herein is determined by such considerations as may be known in the art. The amount should be effective to achieve the desired therapeutic effect described above, depending, among other things, on the type and severity of the disease being treated and the treatment regimen. In some embodiments, the composition comprising pridopidine or a pharma- ceutically acceptable salt thereof is 1-400 mg / day and is administered once a day, twice a day, three times a day, or less frequently than once a day. As is generally known, the effective amount depends on various factors, including various pharmacological parameters such as the affinity of the ligand to the receptor, its distribution profile in the body, its half-life in the body, undesirable side effects (if any), age, and sex, among other factors.

[0086] In some embodiments, pridopidine is administered at a daily dose of 1 mg / day to 400 mg / day. In some embodiments, pridopidine is administered at a daily dose of 1 mg / day to 300 mg / day. In other embodiments, pridopidine is administered at a daily dose of 1 mg / day to 90 mg / day. In other embodiments, pridopidine is administered at a daily dose of 20 mg / day to 90 mg / day. In further embodiments, pridopidine is administered at a daily dose of 45 mg / day to 90 mg / day. In other embodiments, pridopidine is administered at a daily dose of 20 mg / day to 50 mg / day. In further embodiments, pridopidine is administered at a daily dose of 1 mg / day to 10 mg / day. In further embodiments, pridopidine is administered at a daily dose of 10 mg / day to 20 mg / day. In further embodiments, pridopidine is administered at a daily dose of 20 mg / day to 30 mg / day. In a further embodiment, pridopidine is administered in a daily dose of 30 mg / day to 40 mg / day. In a further embodiment, pridopidine is administered in a daily dose of 40 mg / day to 50 mg / day. In a further embodiment, pridopidine is administered in a daily dose of 50 mg / day to 60 mg / day. In a further embodiment, pridopidine is administered in a daily dose of 60 mg / day to 70 mg / day. In a further embodiment, pridopidine is administered in a daily dose of 70 mg / day to 80 mg / day. In a further embodiment, pridopidine is administered in a daily dose of 80 mg / day to 90 mg / day. In a further embodiment, pridopidine is administered in a daily dose of 90 mg / day to 100 mg / day. In a further embodiment, pridopidine is administered in a daily dose of 100 mg / day to 150 mg / day. In a further embodiment, pridopidine is administered in a daily dose of 150 mg / day to 200 mg / day. In a further embodiment, pridopidine is administered in a daily dose of 200 mg / day to 250 mg / day. In a further embodiment, pridopidine is administered in a daily dose of 250 mg / day to 300 mg / day. In a further embodiment, pridopidine is administered in a daily dose of 300 mg / day to 350 mg / day.In a further embodiment, pridopidine is administered in a daily dose of 350 mg / day to 400 mg / day. EXAMPLES

[0087] Example 1: Pridopidine reduced mHtt-induced ER stress

[0088] ER stress was measured in STHdhQ7 / 7 cells transfected with mutant Htt96Q-mCherry (multiplied mutant Htt), which showed visible mutant Htt-mCherry aggregates. The appearance of mutant Htt-mCherry aggregates correlated with high levels of accumulated H2a-GFP, indicative of ER stress (Figure 1). STHdhQ7 / 7 cells expressing Htt20Q-mCherry (wild-type HTT) or Htt96Q-mCherry (mutant HTT) without visible aggregates showed low levels of H2a-GFP (no ER stress).

[0089] Pridopidine significantly reduced H2a-GFP accumulation in mutant Htt aggregate-positive cells in a dose-dependent manner and did not alter H2a-GFP levels in cells that did not contain mHTT aggregates or that expressed Htt20Q-mCherry (Fig. 2 ).

[0090] Thus, pridopidine reduced Htt-induced ER stress in a dose-dependent manner.

[0091] Viral infection-induced ER stress leads to phosphorylation of eIF2α, inhibiting global translation in cells but increasing the expression of UPR-related genes such as ATF4, CHOP, GADD-34, ATF6 and Xbp1 (Bechill et al. 2008; K. Liao et al. 2016).

[0092] Example 2: Pridopidine reduced eIF2α-p levels

[0093] The effect of pridopidine on ER stress was investigated by assessing the phosphorylation levels of the translation initiation factor eIF2α. Phosphorylation of eIF2α is a hallmark of the ER stress response. In Figures 3 and 4, HEK293 cells expressing mutant Htt protein (Htt96Q) show a 1.7-fold and 3.5-fold increase in eIF2α phosphorylation (eIF2α-p) compared to cells expressing wild-type Htt (Htt20Q), respectively. Pridopidine treatment caused a significant dose-dependent decrease in eIF2α phosphorylation (ratio of eIF2α-P / eIF2α) at ​​both 8 h (Figure 3) and 24 h (Figure 4), indicating a decrease in cellular ER stress.

[0094] The effect of pridopidine on ER stress was evaluated in cells where ER stress was not induced by mutant huntingtin but was induced by the potent ER stress inducer thapsigargin. HEK293 cells transfected with Htt20Q-mCherry (wild-type HTT) were treated with 2 μg / ml thapsigargin to induce ER stress. Pridopidine (3 μM) treatment caused a decrease in eIF2α-p (ratio of eIF2α-P / eIF2α) 8 hours after treatment (Figure 5), indicating that pridopidine reduces ER stress regardless of the underlying cause.

[0095] Example 3: Pridopidine reduced eIF2α-p (ER stress) in an S1R-dependent mechanism

[0096] To assess whether the effect of pridopidine on ER stress is S1R-mediated, eIF2α phosphorylation levels were assessed in HEK293 cells genetically deleted for S1R using CRISPR / Cas9 (S1R- / -). ER stress was induced by transfection of mutant Htt (Htt96Q). In S1R+ / + cells, Htt96Q increased ER stress as measured by a two-fold increase in eIF2α-P levels. Pridopidine treatment (0.3 and 3 μM) significantly reduced eIF2α-P levels (Figure 6). Genetic deletion of S1R (S1R- / -) also contributed to increased eIF2α-P levels, implicating S1R in mediating ER stress levels. Pridopidine treatment of Htt96Q-transfected S1R- / - cells had no effect on eIF2α-P levels, indicating that the effects of pridopidine are exquisitely mediated by S1R.

[0097] Levels of the UPR pathway proteins ATF4, CHOP, GADD-34, ATF6 and Xbp1 were upregulated in multiple cellular models in response to infection with coronaviruses (i.e., infectious bronchitis virus (IBV) and mouse hepatitis virus (MHV)) (Y. Liao et al. 2013, Bechill et al. 2008). Bioinformatics analysis predicts a similar effect with SARS-CoV2 (Nabirotchkin et al. 2020).

[0098] Example 4: Pridopidine reduced the levels of UPR PERK pathway markers

[0099] ATF4 translation is increased in response to ER stress and eIF2α phosphorylation, which is part of the UPR pathway and initiates a cascade of transcriptional responses including increased CHOP and GADD-34 translation. The effect of pridopidine on ATF4, CHOP, and GADD-34 protein levels was evaluated in HEK293 cells in which ER stress was induced by mutant Htt (Htt96Q). Htt96Q-induced ER stress increases ATF4 levels by 4.5-fold (Figure 7A), CHOP levels by 5-fold (Figure 7B), and GADD-34 levels by 7-fold (Figure 7C) compared to cells transfected with wild-type Htt (Htt20Q). Pridopidine treatment reduces ATF4, CHOP, and GADD-34 to normal wild-type levels.

[0100] Example 5: Pridopidine reduced the levels of UPR pathway arm ATF6 and IRE markers

[0101] ER stress was evident in HD cells as measured by an increase in two additional UPR pathways, the ATF6 pathway and the IRE1 pathway (increased Xbp1 splicing). In HEK293 cells, ATF6 levels were undetectable in Htt20Q-transfected cells and were therefore increased in mutant Htt96Q-transfected cells. Pridopidine treatment (3 μM) reduced ATF6 levels by 35% (Figure 8). In the IRE1 arm of the UPR pathway, Xbp1s mRNA levels increased by approximately 60%. Pridopidine treatment (3 μM) significantly reduced Xbp1s mRNA levels by 25% (Figure 9). These data indicate that pridopidine reduces ER stress-induced activation of all three arms of the UPR pathway.

[0102] Example 6: Pridopidine reduced mitochondrial ROS production

[0103] HD mouse neurons showed increased sensitivity to oxidative challenge, resulting in increased levels of reactive oxygen species (ROS) and defective antioxidant responses. Striatal neurons from YAC128 HD mice were treated with 1 μM pridopidine followed by induction of ROS production with the mitochondrial respiration inhibitor antimycin A (Ant A). Ant A increased ROS production in YAC128 neurons by approximately two-fold. Pridopidine (1 μM) showed a robust and significant reduction of mitochondrial ROS production to normal levels (Figure 10).

[0104] ATF4 and CHOP are involved in the regulation of mitochondrial function (Sileikyte and Forte 2019). By influencing ATF4 and CHOP levels, as suggested by bioinformatics analysis (Nabirotchkin et al. 2020), SARS-CoV2 may disrupt this regulation to avoid apoptosis, similar to the previously mentioned coronaviruses that were shown to interfere with mitochondrial function (Kim et al. 2018).

[0105] Example 7: SARS-CoV-2 infection reduces mitochondrial membrane potential

[0106] The Nsp10 protein of SARS-CoV directly interacts with cytochrome oxidase II, a component of mitochondrial complex IV. This interaction leads to a decrease in the activity of cytochrome oxidase and loss of mitochondrial membrane potential. Taken together, SARS-CoV-2 affects the mitochondrial redox enzyme system and reduces the mitochondrial membrane potential (Q. Li et al. 2005).

[0107] Example 8: Pridopidine increases mitochondrial membrane potential

[0108] The role of S1R in regulating mitochondrial membrane potential (MMP) was investigated in striatal neurons from wild-type (WT) controls and YAC128 HD mice. HD neurons show reduced MMP compared to WT. In WT neurons, pridopidine caused a significant increase in MMP at 1 μM and a trend towards increased MMP at 0.1 μM. In HD neurons, where MMP was reduced by 25% compared to WT, both doses of pridopidine caused a significant increase in MMP. Thus, pridopidine rescues the membrane potential impairment (Figure 11).

[0109] Example 9: The effect of pridopidine on mitochondrial membrane potential is mediated by S1R

[0110] To confirm that the effects of pridopidine are mediated by S1R, we knocked down S1R in HD lymphoblasts, achieving an approximately 83% reduction in protein levels. 2 O 2 Treatment significantly reduced MMP in both S1R+ / + and S1R-KD cells by 25% and 75%, respectively. Pridopidine treatment (5 μM) fully restored MMP in S1R+ / + cells, but not in S1R KD cells. Thus, pridopidine has a protective effect against MMP. 2 O 2 The effect of pridopidine on challenge-induced MMP reduction was abolished in cells with reduced S1R levels (p<0.001), indicating that the pridopidine effect was S1R-dependent (Figure 12).

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Claims

1. 1. A composition for treating, reducing the incidence, suppressing or inhibiting a viral infection, disease, disorder, or symptoms thereof in a subject, comprising: A composition comprising pridopidine, or a pharma- ceutically acceptable salt thereof.

2. 2. The composition of claim 1, wherein the viral infection, disease, or disorder comprises a human coronavirus, SARS, MERS coronavirus, SARS coronavirus 2, or a mutant therefrom.

3. The composition of claim 2, wherein the disease is COVID-19.

4. The composition of any one of claims 1 to 3, further reducing ER stress in the subject.

5. 4. The composition of claim 3, wherein the COVID-19 symptoms include kidney failure, fever, fatigue, dry cough, body aches, nasal congestion, runny nose, sore throat, diarrhea, or a combination thereof.

6. The composition according to any one of claims 1 to 5, further comprising a combination of at least one of analogues 1 to 7 or a salt thereof. 【Chemistry 1】

7. The composition, Pridopidine or a pharma- ceutically acceptable salt thereof; The composition of claim 6 comprising a combination with Compound 1 or a pharma- ceutically acceptable salt thereof.

8. The composition comprising: Pridopidine or a pharma- ceutically acceptable salt thereof; The composition of claim 6 comprising in combination with compound 4 or a pharma- ceutically acceptable salt thereof.

9. The composition, Pridopidine or a pharma- ceutically acceptable salt thereof; Compound 1 or a pharma- ceutically acceptable salt thereof; The composition of claim 6 comprising in combination with compound 4 or a pharma- ceutically acceptable salt thereof.

10. The composition according to any one of claims 1 to 9, wherein the pridopidine is in base form or in a pharma- ceutically acceptable salt form.

11. The pridopidine salts are selected from the group consisting of pridopidine hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, isonicotinate, acetate, lactate, salicylate, citrate, D,L-tartrate, L-tartrate, D-tartrate, pantothenate, bitartrate, ascorbate, succinate, hemisuccinate, maleate, gentisate, gentisic acid, fumarate, gluconate, and glutamate.

10. The composition of any one of claims 1 to 9, comprising a chlorate, glycolate, saccharate, formate, besylate, benzoate, glutamate, malate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, oxalate, tosylate, naphthalene-2-sulfate, or pamoate (i.e., 1,1'-methylene-bis-(2-hydroxy-3-naphthoic acid)) salt.

12. 1. A composition for reducing endoplasmic reticulum stress (ER stress) resulting from a viral infection, disease, or disorder in a subject, comprising: A composition comprising pridopidine, or a pharma- ceutically acceptable salt thereof.

13. 13. The composition of claim 12, wherein the viral infection, disease, or disorder comprises a human coronavirus, SARS, MERS coronavirus, SARS coronavirus 2, and variants therefrom.

14. The composition of claim 12 or 13, wherein the disease is COVID-19.

15. The composition according to any one of claims 12 to 14, wherein the composition is in combination with at least one of analog compounds 1 to 7 or a salt thereof. 【Chemistry 2】

16. The composition, A composition comprising pridopidine or a pharma- ceutically acceptable salt thereof; 16. The composition of claim 15, comprising a combination with Compound 1 or a pharma- ceutically acceptable salt thereof.

17. The composition comprising: Pridopidine or a pharma- ceutically acceptable salt thereof; 16. The composition of claim 15, comprising in combination with compound 4, or a pharma- ceutically acceptable salt thereof.

18. The composition, Pridopidine or a pharma- ceutically acceptable salt thereof; Compound 1 or a pharma- ceutically acceptable salt thereof; 16. The composition of claim 15, comprising in combination with compound 4, or a pharma- ceutically acceptable salt thereof.

19. The composition according to any one of claims 15 to 18, wherein the pridopidine is in base form or in a pharma- ceutically acceptable salt form.

20. The pridopidine salts may be pridopidine hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, isonicotinate, acetate, lactate, salicylate, citrate, D,L-tartrate, L-tartrate, D-tartrate, pantothenate, bitartrate, ascorbate, succinate, hemisuccinate, maleate, gentisate, gentisic acid, fumarate, gluconate, gluconate, or the like.

20. The composition of any one of claims 15 to 19, comprising a ronate, glycolate, saccharate, formate, besylate, benzoate, glutamate, malate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, oxalate, tosylate, naphthalene-2-sulfate, or pamoate (i.e., 1,1'-methylene-bis-(2-hydroxy-3-naphthoic acid)) salt.

21. The composition of any one of claims 1 to 20, wherein the composition is administered by systemic administration.

22. The composition of claim 21 , wherein the composition is administered by oral administration.

23. The composition of any one of claims 1 to 22, wherein the composition is formulated as an oral liquid, solid, semi-solid dosage form, injectable solution, dermal / transdermal dosage form, ophthalmic dosage form, inhalable composition.

24. 24. The composition of claim 23, wherein the composition is formulated as an inhalable powder, injectable, liquid, gel, solid, capsule, eye drop, or tablet.

25. 25. The composition of any one of claims 1 to 24, wherein the composition is administered once a day, twice a day, three times a day, or less than once a day.

26. 26. The composition of claim 25, wherein the composition is administered in one dose, two doses, or three doses per day.

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