Responder selection and treatment with sigma-1 receptor therapy

Abstract

The present disclosure provides genes and genetic polymorphisms associated with an altered response of a subject to Sigma-1 receptor therapy and methods of determining if a subject is responsive or non-responsive to Sigma-1 receptor therapy. The methods can be used to optimize treatment for subjects in need of Sigma-1 receptor therapy. Based on the predicted responsiveness of a subject to the therapy, personalized treatment can be provided to each subject.

Description

PATENT Attorney Docket No.: 085102-875342Via Patent Center RESPONDER SELECTION AND TREATMENT WITH SIGMA-1 RECEPTOR THERAPYCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of U. S. Provisional Patent Application No. 63 / 791,216 filed April 18, 2025, entitled “RESPONDER SELECTION AND TREATMENT WITH SIGMA-1 RECEPTOR THERAPY,” and the entire disclosure of which is hereby incorporated by reference.FIELD OF THE DISCLOSURE

[0002] The present disclosure relates generally to methods of determining if a subject is or will be responsive or non-responsive to Sigma-1 receptor therapy.BACKGROUND

[0003] Alzheimer’s disease (AD) is a progressive neurodegenerative disorder characterized by loss of memory and other cognitive functions, leading to interference in daily life activities. At present, there is a dearth of pharmacological treatments that have been shown to alter progression of the disease. Sigma-1 receptor (SIGMAR1), a modulator of calcium homeostasis and mitochondrial function, is a new target relevant to conditions variously known as neurodegenerative and neurodevelopmental disorders or indications, including AD. SIGMAR1 activation has been shown to reduce key pathophysiological processes in AD including hyperphosphorylation of tau, neuroprotection, neurodegeneration, and oxidative stress. Activation of SIGMAR1 also leads to increase in autophagic flux in human cells and in vivo. However, genetic heterogeneity of the AD patient population and lack of objective efficacy measures or predictive biomarkers of response to treatment contribute to severe limitations in treatment efficacy. This heterogeneity is a background noise that can obscure successful treatments.108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center

[0004] Therefore, there is a need for predictive biomarkers and methods for developing personalized protocols for treating patients in need of Sigma-1 receptor therapy.SUMMARY OF THE DISCLOSURE

[0005] One aspect of the present disclosure encompasses a method of selecting a subject responsive to Sigma-1 receptor therapy (e.g., Sigma-1 receptor agonist therapy). The method comprises obtaining or having obtained the results of a test of a biological sample from the subject which determines the presence of at least one polymorphism selected from the group consisting of APOE4 (rs429358 and / or rs7412), COL24A1 (rs60891279), ADRA1D (re184418609), ANXA2(rs17845226), SIGMAR1_Q2P, COMT_L146fs, KANSL1_P1010L / P304L / P946L, DHCR7_M220T, HLA-DRB1_ A244T, HLA-DRB1_ S66Y, HLA-DRB1_ Y89S, MS4A6E_M59T, RIN3_ H215R, DPYD_I543V, and combinations thereof in the subject. The method further comprises excluding the subject as non-responsive to Sigma-1 receptor therapy if one or more of the polymorphism(s) is / are present in the subject.

[0006] Another aspect of the present disclosure encompasses a method of selecting subjects responsive to Sigma-1 receptor therapy, such as Sigma-1 receptor agonist therapy. The method comprises detecting or having detected whether at least one polymorphism is present at a genetic locus in a biological sample obtained from the subject by sequencing the genetic locus comprising the polymorphism. The method further comprises excluding the subject as non-responsive to Sigma-1 receptor therapy if a polymorphism is present in the subject the subject. In some aspects, the at least one polymorphism is selected from the group consisting of APOE4 (rs429358 and / or rs7412), COL24A1 (rs60891279), ADRA1D (re184418609), ANXA2 (rs17845226), SIGMAR1_Q2P, COMT_L146fs, KANSL1_P1010L / P304L / P946L, DHCR7_M220T, HLA-DRB1_ A244T, HLA-DRB1_ S66Y, HLA-DRB1_ Y89S, MS4A6E_M59T, RIN3_ H215R, DPYD_I543V, and combinations thereof.2108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center

[0007] Yet another aspect of the present disclosure encompasses a method of selecting subjects responsive to Sigma-1 receptor therapy (e.g., Sigma-1 receptor agonist therapy). The method comprises obtaining or having obtained the results of a test of a biological sample from the subject which determines the level of expression of RNA encoded by at least one gene selected from the group consisting of APOE4, COL24A1, ADRA1 D, ANXA2, SIGMAR1, COMT, KANSL1, DHCR7, HLA-DRB1, MS4A6E, RIN3, DPYD, and combinations thereof, and comparing the level of the RNA in the test sample to the level of the RNA in a subject responsive to Sigma-1 receptor therapy. A level of the RNA in the test sample substantially similar to the level of the RNA in the subject responsive to Sigma-1 receptor agonist therapy identifies a subject responsive to Sigma-1 receptor therapy.

[0008] Another aspect of the present disclosure encompasses a method of selecting subjects responsive to Sigma-1 receptor therapy (e.g., Sigma-1 receptor agonist therapy). The method comprises determining or having determined the level of expression of RNA encoded by at least one gene selected from the group consisting of APOE4, COL24A1, ADRA1 D, ANXA2, SIGMAR1, COMT, KANSL1, DHCR7, HLA-DRB1, MS4A6E, RIN3, DPYD, and combinations thereof. The method also comprises excluding the subject as non-responsive to Sigma-1 receptor therapy if the level of the RNA in the test sample is substantially different from the level of the RNA in a subject responsive to Sigma-1 receptor therapy.

[0009] Another aspect of the present disclosure encompasses a method of treating a subject in need of Sigma-1 receptor therapy, such as Sigma-1 receptor agonist therapy. The method comprises obtaining or having obtained the results of a test of a biological sample from the subject which detects the presence of at least one polymorphism selected from the group consisting of APOE4 (rs429358 and / or rs7412), COL24A1 (rs60891279), ADRA1D (re184418609), ANXA2 (rs17845226), SIGMAR1_Q2P, COMT_L146fs, KANSL1_P1010L / P304L / P946L, DHCR7_M220T, HLA-DRB1_ A244T, HLA-DRB1_ S66Y, HLA-DRB1_ Y89S, MS4A6E_M59T, RIN3_ H215R, DPYD_I543V, and combinations thereof in the subject. The method also comprises administering a therapeutically effective amount of a Sigma-1 receptor agonist to the subject if a polymorphism(s) is / are not present in the subject.3108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center

[0010] Another aspect of the present disclosure encompasses a method of treating a subject in need of Sigma-1 receptor therapy (e.g., Sigma-1 receptor agonist therapy). The method comprises obtaining or having obtained the results of a test of a biological sample from the subject which determines the level of expression of RNA encoded by at least one gene selected from the group consisting of APOE4, COL24A1, ADRA1 D, ANXA2, SIGMAR1, COMT, KANSL1, DHCR7, HLA-DRB1, MS4A6E, RIN3, DPYD, and combinations thereof. The method further comprises comparing the level of the RNA in the test sample to the level of the RNA in a subject responsive to Sigma-1 receptor therapy, and administering a therapeutically effective amount of a Sigma-1 receptor agonist to the subject if a level of the RNA in the test sample is substantially similar to the level of the RNA in a subject responsive to Sigma-1 receptor therapy.

[0011] Yet another aspect of the present disclosure encompasses a method of detecting the presence of at least one polymorphism associated with response to Sigma-1 receptor therapy (e.g., Sigma-1 receptor agonist therapy) in a subject. The method comprises obtaining or having obtained a biological sample from the subject and detecting the polymorphism in the biological sample by evaluating the sequence of a genetic locus comprising the polymorphism. In some aspects, the at least one polymorphism is selected from the group consisting of APOE4 (rs429358 and / or rs7412), COL24A1 (rs60891279), ADRA1D (re184418609), ANXA2 (rs17845226), SIGMAR1_Q2P, COMT_L146fs, KANSL1_P1010L / P304L / P946L, DHCR7_M220T, HLA-DRB1_ A244T, HLA-DRB1_ S66Y, HLA-DRB1_ Y89S, MS4A6E_M59T, RIN3_ H215R, DPYD_I543V, and combinations thereof in the subject.

[0012] In various aspects, the Sigma-1 receptor therapy involves administering to the subject a Sigma-1 receptor agonist alone or in combination with one or more additional pharmaceutical substances (including, but not limited to, additional or alternative Sigma-1 receptor agonists). In various embodiments, the Sigma-1 receptor agonist is selected from: tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride (A2-73), tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine (A2-73 freebase), tetrahydro-N, N-dimethyl-5,5-diphenyl-3-furanmethanamine hydrochloride (A1-41); tetrahydro-N, N-dimethyl-5,5-diphenyl-3-furanmethanamine (A1-41 freebase); 1-(2,2-diphenyltetrahydrofuran-3-yl)-N-4108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center methylmethanamine hydrochloride (A19-144); 1-(2,2-diphenyltetrahydrofuran-3-yl)-N-methylmethanamine (A19-144 freebase); (1 -(2,8-dimethyl- l-thia-3,8-diazaspiro(4.5)dec-3-yl)-3-(IH-indol-3-yl)propan-l-one) (A3-71); an enantiomer thereof, a crystal form thereof, a co-crystal form thereof, a pharmaceutically acceptable salt thereof, a crystal or co-crystal of the pharmaceutically acceptable salt thereof, or any combination thereof. In yet another aspect, the Sigma-1 receptor agonist is selected from: tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine in amorphous form (A2-73 freebase amorphous form); tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine in crystal Form I (A2-73 Freebase Crystal Form I); tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride in crystal Form I (A2-73 Crystal Form I); tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride in crystal Form II (A2-73 Crystal Form II); tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride in crystal Form III (A2-73 Crystal Form III); (+) tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride in amorphous form ((+) A2-73 amorphous form); (-) tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride in amorphous form ((-) A2-73 amorphous form); (+) tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride in a crystal form ((+) A2-73 crystal form); (-) tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride in a crystal form ((-) A2-73 crystal form); tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrogen fumarate in amorphous form (A2-73 hydrogen fumarate amorphous form); tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrogen fumarate in crystal Form I (A2-73 hydrogen fumarate Crystal Form I); tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrogen fumarate in crystal Form II (A2-73 hydrogen fumarate Crystal Form II); tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrogen fumarate in crystal Form III (A2-73 hydrogen fumarate Crystal Form III); tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrogen fumarate in crystal Form IV (A2-73 hydrogen fumarate Crystal Form IV); tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrogen fumarate in crystal Form V (A2-73 hydrogen fumarate Crystal Form V); tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine mesylate in crystal Form I (A2-73 mesylate Crystal Form I); tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine sulfate in crystal Form I (A2-73 sulfate Crystal Form I);5108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine sulfate in crystal Form II (A2-73 sulfate Crystal Form II); tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine oxalate in crystal Form I (A2-73 oxalate Crystal Form I); tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine oxalate in crystal Form II (A2-73 oxalate Crystal Form II); tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine dihydrogen phosphate in crystal Form I (A2-73 dihydrogen phosphate Crystal Form I); tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine edisylate in crystal Form I (A2-73 edisylate Crystal Form I); tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine benzoate in crystal Form I (A2-73 benzoate Crystal Form I); tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrobromide in crystal Form A (A2-73 hydrobromide Crystal Form A); tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrobromide in crystal Form B (A2-73 hydrobromide Crystal Form B); tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine maleate in crystal Form S5 (A2-73 maleate Crystal Form S5); tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine maleate in crystal Form S6 (A2-73 maleate Crystal Form S6); co-crystal Form CSII of tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride with tartaric acid (A2-73 co-crystal with tartaric acid Crystal Form CSII); co-crystal Form CSII I of tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride with citric acid (A2-73 co-crystal with citric acid Crystal Form CSIII); co-crystal Form CSIV of tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride with malic acid (A2-73 co-crystal with malic acid Crystal Form CSIV); a co-crystal of tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride with zinc chloride (A2-73 co-crystal with zinc chloride); a co-crystal of (-) tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride with zinc chloride ((-) A2-73 co-crystal with zinc chloride); a co-crystal of (+) tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride with zinc chloride ((+) A2-73 co-crystal with zinc chloride); and any combination thereof. In another aspect, the Sigma-1 receptor agonist therapy comprises an oral composition comprising A2-73 free base, A2-73 amorphous form, A2-73 Crystal Form I, A2-73 Crystal Form II, A2-73 Crystal Form III, (-) A2-73 enantiomer, (+) A2-73 enantiomer, or any combination thereof. In yet another aspect, the composition comprises about 5 to 50 mg of A2-73 free base or A2-73 Crystal Form I (for example, about 5 mg, about 10 mg, about 20 mg, about6108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center 30 mg, about 40 mg, or about 50 mg). In yet another aspect, the composition is an oral liquid and comprises A2-73 in an amount of about 5 mg, about 10 mg, about 20 mg, or about 30 mg.

[0013] In various aspects, the administering or administration comprises a route of administration selected from the group consisting of oral administration, subcutaneous injection, intravenous injection, intraocular injection, intradermal injection, intramuscular injection, intraperitoneal injection, intratracheal administration, inhalation, intranasal administration, sublingual administration, buccal administration, rectal administration, vaginal administration, topical administration, and transdermal administration. In a particular aspect, the administering or administration is oral administration. In another particular aspect, the administering or administration is intravenous administration. In another particular aspect, the administering or administration is transdermal administration.

[0014] In various aspects, the test biological sample or the control biological sample each independently comprises a blood serum sample, a blood plasma sample, or a sample suitable for transcriptomic analysis. In one aspect, the test biological sample and the control biological sample are the same type of biological samples. In another aspect, the test biological sample and the control biological sample are different types of biological samples.

[0015] In various aspects, the treatment period or the trial period is about 12 weeks or less. In another aspect, the treatment period is about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 9 weeks, about 10 weeks, about 11 weeks, about 12 weeks. Yet in another aspect, the Sigma-1 receptor agonist is administered daily. In various aspects, the Sigma-1 receptor agonist is tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine in crystal Form I, or the Sigma-1 receptor agonist is tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride in Crystal Form I. In various aspects, the Sigma-1 receptor agonist is administered daily in an amount of about 5 mg to about 60 mg. In various aspects, the Sigma-1 receptor agonist is administered in an oral liquid comprising up to about 30 mg of the Sigma-1 receptor agonist. In various aspects, the Sigma-1 receptor agonist comprises tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine or tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine7108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center hydrochloride in an oral solution, and wherein the tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine or the tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride is administered once daily for a period of 12 weeks in an escalating dose regimen starting at about 30 mg and ending at about 50 mg. In some aspects, the Sigma-1 receptor agonist comprises tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine or tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride in an oral solution, and wherein the tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine or the tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride is administered once daily for a period of 12 weeks in an escalating dose regimen starting at about 5 mg and ending at about 30 mg (for example, an escalating dose of 5 mg, 10 mg, 20 mg, and 30 mg).REFERENCE TO COLOR FIGURES

[0016] The application file contains at least one photograph executed in color. Copies of this patent application publication with color photographs will be provided by the Office upon request and payment of the necessary fee.BRIEF DESCRIPTION OF THE FIGURES

[0017] FIG. 1. An illustrative overview of study design, data availability and analytical methods. FIG. 1 A: Summary of consecutive clinical trials involving two parts: Part A over 5 weeks and Part B over 52 weeks (57-week study), and 208-week study extension study over a planned 208 week period following immediately after the initial trial, totaling in a cumulative 265-weeks. FIG. 1B: Summary of integrated data sources. FIG. 1 C: Description of number of patient descriptors incorporated in the two analytical steps applied in the study 1) Unsupervised FCA rule-based analysis of response at week 57, 2) Longitudinal confirmation using Mixed Effect modelling of response over 148 weeks incorporating markers found at week 57 to model two groups of ANAVEX2-73 concentrations.

[0018] FIGS. 2A-2C. Schematic illustrations of dosages and plasma concentration of ANAVEX2-73 and ANAVEX19-144. FIG. 2A: Dose administration cross-over design including eight possible administration schemes, where a single 8108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center dose regime is given in each period (Intravenous (IV) or Orally (Oral)), either beginning with an IV or Oral administration with two possible doses allocated randomly. FIG. 2B: Pharmacokinetics parameter area under curve between 0 and infinity (AUCO-inf) of ANAVEX2-73 after first administration (24h first hours of the trial) in 57-week study Part A with different randomized administration dosages. FIG. 2C: Boxplot of mean concentration groups of ANAVEX2-73 in the plasma.

[0019] FIG. 3. 3D crystal structure of SIGMAR1 visualized using VistaPDB 1.0 software.

[0020] FIGS. 4A-4B. Plots depicting MMRM-LME models of MMSE and ADCS-ADL change from baseline over 148 weeks. FIG. 4A: MMRM-LME-adjusted slopes for the high concentration (Green) cohort versus low and medium concentration patient cohort (Magenta) with time (in weeks) against adjusted Delta MMSE. Average adjusted values with residuals at the population level were plotted at each time point (dotted line). FIG.4B: MMRM-LME-adjusted slopes for the high concentration (Green) cohort versus low and medium concentration patient cohort (Magenta) with time (in weeks) against adjusted Delta MMSE. Average adjusted values with residuals at the population level were plotted at each time point (dotted line).

[0021] FIG. 5. Plot of unadjusted values of Delta ADCS-ADL over 148 weeks. The plot presents (unadjusted) mean trajectories of Delta ADCS-ADL score through interim 148 weeks of subgroups of patients in the 208-week study depending on biomarker status (absent or present) and of patients given standard of care. The subgroups of patients in the 208-week study are represented (blue, green, orange, pink, purple and turquoise) plot depending on identified biomarker and baseline criteria characteristics.

[0022] FIGS. 6A-6B. Plots showing the therapeutic response showing Longitudinal Delta MMSE and Delta ADCS-ADL over 148 week of combined studies. The patient presented in FIG. 6A and FIG. 6B has SIGMAR1 gene wild type, mean high concentration levels of ANAVEX2-73 in plasma in Part B and baseline MMSE >20.9108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center

[0023] FIGS. 7A-7B. Plots showing that the presence of SIGMAR1 Pro2 mutation is associated with a poor response to a sigma-1 agonist as measured by both MMSE (FIG. 7A) and ADCS-ADL deltas (FIG. 7B).

[0024] FIGS. 8A-8F. FIG. 8A: Plot showing that the frameshift COMT Leu 146 mutation is strongly linked to a high ADCS-ADL Delta from baseline (BL). FIG. 8B:Plot showing that the absence of KANSL1 Pro304 / 946 / 1010 mutation is strongly linked to a high ADCS-ADL Delta from BL. FIG. 8C: Plot showing that the absence of SIGMAR1 Pro2 mutation is strongly linked to a high ADCS-ADL Delta from BL.FIG. 8D: Plot showing that the absence of frameshift COMT Leu146 mutation is strongly linked to a high MMSE delta from BL. FIG. 8E: Plot showing that the absence of KANSL1 Pro304 / 946 / 1010 mutation is strongly linked to a high MMSE delta from BL. FIG. 8F: Plot showing that the absence of SIGMAR1 Pro2 mutation is strongly linked to a high MMSE delta from BL.

[0025] FIG. 9. A plot showing high SIGMAR1 RNA expression is strongly linked to high ADCS-ADL slope values from BL (improvement). The analysis includes data of 100% of all patients (N=21). TPM means transcripts per kilobase million.

[0026] FIGS. 10A-10D provide 243 genes selected from the 27,155 mapped sequences obtained from NGS analysis (shown in FIGS. 10A-10C). FIG. 10A lists the 102 genes selected on the basis of their involvement in neurodegenerative diseases. FIG. 10B provides the added 113 genes of the cytochrome P450s gene family and 10 genes from the methyltransferase gene family. FIGS. 10C-10D shows the 20 genes selected as part of SIGMARTs functional interactome, based on a confidence score of 0.150 obtained from the STRING database (January 2018).

[0027] FIG. 11. A description of Anavex’s AD-004 Phase IIB / III Early Alzheimer’s Disease Trial.

[0028] FIGS. 12A-12G. Plots showing that the SIGMAR1 / COL24A1 doublenegative population experiences increased efficacy with A3-73 therapy; in particular, 75.9% less decline (p<0.001 ). MMRM analysis of change in ADAS-Cog13 total score from baseline for the total (m)ITT population, or to subgroups selecting for GWAS-identified gene variant populations. Numbers of observations below the plots10108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center represent all participants with non-missing questionnaire and covariate data at each time point. Values represent the least-squares mean at each analysis visit, with error bars representing standard error (SE). *: p < 0.05; **: p < 0.01; ***: p < 0.001; ****: p < 0.0001.

[0029] FIGS. 13A-13G Plots showing that the SIGMAR1 WT / COL24A1 WT (+ / +) population experiences 38.8% less decline in the active blarcamesine group (p<0.001 ). MMRM analysis of change in ADCS-ADL total score from baseline for the total (m)ITT population, or to subgroups selecting for GWAS-identified gene variant populations. Numbers of observations below the plots represent all participants with non-missing questionnaire and covariate data at each time point. Values represent the least-squares mean at each analysis visit, with error bars representing standard error (SE). *: p < 0.05; **: p < 0.01; ***: p < 0.001; **** p < 0.0001.

[0030] FIG. 14A-14G. Plots showing that the SIGMAR1 WT / COL24A1 WT (+ / +) population shows 57.0% less decline in the active blarcamesine group (p<0.001 ). MMRM analysis of change in CDR-SB score from baseline for the total (m)ITT population, or to subgroups selecting for GWAS-identified gene variant populations. Numbers of observations below the plots represent all participants with non-missing questionnaire and covariate data at each time point. Values represent the least-squares mean at each analysis visit, with error bars representing standard error (SE). *: p < 0.05; ** p < 0.01; ***: p < 0.001; **** p < 0.0001.

[0031] FIG. 15A-15G. Plots showing that the SIGMAR1 WT / COL24A1 WT (+ / +) population experiences improved blarcamesine efficacy, less than 0.5 point decline at Wk48 (p<0.001). MMRM analysis of CGI-Improvement score from baseline for the total (m)ITT population, or to subgroups selecting for GWAS-identified gene variant populations. Numbers of observations below the plots represent all participants with non-missing questionnaire and covariate data at each time point. Values represent the least-squares mean at each analysis visit, with error bars representing standard error (SE). *: p < 0.05; **: p < 0.01; ***: p < 0.001; ****: p < 0.0001.

[0032] FIG. 16A-16G. Plots showing that the SIGMAR1 WT / COL24A1 WT (+ / +) population experiences improved blarcamesine efficacy, 48.3% less decline at Wk48 (p=0.0052). MMRM analysis of change in MMSE score from baseline for the 11108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center total (m)ITT population, or to subgroups selecting for GWAS-identified gene variant populations. Numbers of observations below the plots represent all participants with non-missing questionnaire and covariate data at each time point. Values represent the least-squares mean at each analysis visit, with error bars representing standard error (SE). *: p < 0.05; ** p < 0.01; *** p < 0.001; p < 0.0001.

[0033] FIG. 17A-17G. Plots showing that the SIGMAR1 WT / COL24A1 WT (+ / +) population experiences improved blarcamesine efficacy, 66.1% less decline at Wk48 (p=0.0001) and also that SIGMAR1 WT (+ / +) and COL24A1 WT (+ / +) have additive benefits to blarcamesine response. MMRM analysis of change in NPI-Q score from baseline for the total (m)ITT population, or to subgroups selecting for GWAS-identified gene variant populations. Numbers of observations below the plots represent all participants with non-missing questionnaire and covariate data at each time point. Values represent the least-squares mean at each analysis visit, with error bars representing standard error (SE). *: p < 0.05; **: p < 0.01; ***: p < 0.001; ****: p < 0.0001.

[0034] FIG. 18A-18G. Plots showing that Patient-assessed QoL-AD is significantly improved for SIGMAR1 WT I COL24A1 WT (+ / +) in the blarcamesine group (p=0.0150). MMRM analysis of change in Patient QoL-AD score from baseline for the total (m)ITT population, or to subgroups selecting for GWAS-identified gene variant populations. Numbers of observations below the plots represent all participants with non-missing questionnaire and covariate data at each time point. Values represent the least-squares mean at each analysis visit, with error bars representing standard error (SE). *: p < 0.05; **: p < 0.01; ***: p < 0.001; ****: p < 0.0001

[0035] FIG. 19A-19G. Plots showing that Caregiver-assessed QoL-AD is significantly improved for COL24A1 WT (+ / +) in the blarcamesine group (p=0.0366). MMRM analysis of change in caregiver QoL-AD score from baseline for the total (m)ITT population, or to subgroups selecting for GWAS-identified gene variant populations. Numbers of observations below the plots represent all participants with non-missing questionnaire and covariate data at each time point. Values represent the least-squares mean at each analysis visit, with error bars representing standard error (SE). *: p < 0.05; p < 0.01; ***: p < 0.001; ****: p < 0.0001.12108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center

[0036] FIG. 20. Schematic outline of study of Example 6 (below), workflow, and methodology.

[0037] FIG. 21A-FIG. 21 B. FIG. 21A illustrates Pharmacogenomics GWAS for the blarcamesine response for outcome CDR-SB at timepoint 48 weeks, Manhattan plot, right-hand side SNPs with -logw[p]>5. FIG. 21B illustrates Pharmacogenomics GWAS for the blarcamesine response for outcome ADAS-Cog13 at timepoint 48 weeks, Manhattan plot, right-hand side SNPs with -logi o[p]>5, plus highlighted by red circle, SNP rs60891279, COL24A1(R541H).

[0038] FIG. 22. COL24A1 gene expression in human brain. COL24A1 gene expression in human brain for spatial context of the projected pharmacogenomic implication of COL24A1 genetic variation in the blarcamesine response. COL24A1 mRNA expression in regional brain samples from two cohorts, aging controls (n=51) vs dementia (n=43), from the Allen Brain Atlas human brain gene expression repository. Left-hand diagram indicates upregulated expression in hippocampi, appreciable expression in cortex, cortical expression overall higher than in white matter, note absence of significant differences between dementia and control cohorts. Right-hand side shows robust hippocampal expression also in a subgroup of neuropathologically more severe dementia cases (Braak4-6, Cerad 2-3; n=25), statistically not significantly different from all-dementia (n=43) and from healthy aging controls (n=51).

[0039] FIG. 23. Patho-mechanistic schematic. Schematic how COL24A1 with its WT allele versus its missense mutation, arginine641— >histidine, could influence the response to blarcamesine, as it enhances neural autophagy in early Alzheimer's disease.DETAILED DESCRIPTION

[0040] The present disclosure is based in part on the discovery of methods of determining if a subject is responsive or non-responsive to Sigma-1 receptor therapy. The methods can comprise detecting the presence of a gene(s) and / or a polymorphism(s) associated with an altered response ( / .e., responsiveness or nonresponsiveness) to Sigma-1 receptor therapy. The method can also comprise13108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center determining the level of expression of RNA encoded by a gene associated with an altered response ( / .e., responsiveness or non-responsiveness) to Sigma-1 receptor therapy. The methods can be used to optimize treatment for subjects in need of Sigma-1 receptor therapy. In some aspects, the Sigma-1 receptor therapy is a Sigma-1 receptor agonist therapy. Based on the predicted responsiveness of a subject to the therapy, personalized treatment can be provided to each subject. For instance, responsive subjects can be selected for treatment using a Sigma-1 receptor agonist, whereas non-responsive subjects may be provided alternative, more appropriate treatment regimens early in the disease development process and / or even before treatment is started. This is in contrast to current methods wherein a subject is determined to be responsive or non-responsive after an extended period of treatment. The genes, polymorphisms, methods of identifying the genes and / or polymorphisms, and methods of using the gene and / or polymorphism information are described below.I. Genes and / or Polymorphisms Associated with Response of a Subject to Sigma-1 Receptor Therapy

[0041] One aspect of the present invention encompasses genes and / or genetic polymorphisms predictive of increased or reduced (or zero) therapeutic response to treatment using Sigma-1 receptor therapy (e.g., Sigma-1 receptor agonist therapy). Genes of interest can be at any chromosome in a subject provided the gene is associated with a positive response of a subject to Sigma-1 receptor therapy. Similarly, polymorphisms of interest can be at any genetic locus in the genome in a subject provided the polymorphism is associated with a positive response of a subject to Sigma-1 receptor therapy. For instance, the polymorphism can be in a coding region or noncoding region of the genome of the subject, including mitochondrial DNA. The polymorphism(s) can result from an insertion, a deletion, or a single nucleotide polymorphism (SNP) in a gene or genetic locus.

[0042] In some aspects, the gene(s) associated with response to Sigma-1 receptor therapy (e.g., Sigma-1 receptor agonist therapy) is / are in a gene selected from APOE4, COL24A1, ADRA1 D, ANXA2, SIGMAR1, COMT, KANSL1, DHCR7, HLA-DRB1, MS4A6E, RIN3, GAMT, IL10, MTUS1, PPARG, and combinations 14108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center thereof. In one aspect, the gene is / are selected from APOE4, COL24A1, ADRA1 D, ANXA2, SIGMAR1, COMT, KANSL1, DHCR7, HLA-DRB1, MS4A6E, RIN3, and combinations thereof. In another aspect, the gene is / are selected from APOE4, COL24A1, ADRA1D, ANXA2, SIGMAR1, COMT, KANSL1, and combinations thereof. In another aspect, the gene is / are selected from APOE4, COL24A1, ADRA1D, ANXA2, SIGMAR1, COMT, and combinations thereof. In another aspect, the gene is / are selected from APOE4, COL24A1, ADRA1 D, and ANXA2. In another aspect, the gene is APOE4. In another aspect, the gene is COL24A1. In another aspect, the gene is ADRA1 D. In another aspect, the gene is ANXA2. In another aspect, the gene is SIGMAR1. In another aspect, the gene is COMT. In another aspect, the gene is KANSL1. In yet another aspect, two genes are involved and one gene is selected from APOE4, COL24A1, ADRA1 D, ANXA2, SIGMAR1, COMT, KANSL1 and the second gene is a different gene selected from APOE4, COL24A1, ADRA1 D, ANXA2, SIGMAR1, COMT, KANSL1. In yet another aspect, three genes are involved and the three different genes are selected from APOE4, COL24A1, ADRA1D, ANXA2, SIGMAR1, COMT, KANSL1.

[0043] In some aspects, the polymorphism(s) associated with response to Sigma-1 receptor therapy (e.g., Sigma-1 receptor agonist therapy) is / are in a gene selected from APOE4, COL24A1, ADRA1D, ANXA2, SIGMAR1, COMT, KANSL1, DHCR7, HLA-DRB1, MS4A6E, RIN3, GAMT, IL10, MTUS1, PPARG, and combinations thereof. In one aspect, the polymorphism is in a gene selected from APOE4, COL24A1, ADRA1D, ANXA2, SIGMAR1, COMT, KANSL1, DHCR7, HLA-DRB1, MS4A6E, RIN3, and combinations thereof. In another aspect, the polymorphism is in a gene selected from APOE4, COL24A1, ADRA1D, ANXA2, SIGMAR1, COMT, KANSL1, and combinations thereof. In another aspect, the polymorphism is in a gene selected from APOE4, COL24A1, ADRA1D, ANXA2, SIGMAR1, COMT, and combinations thereof. In another aspect, the polymorphism is in a gene selected from APOE4, COL24A1, ADRA1D, ANXA2, and SIGMAR1. In another aspect, the polymorphism is in the gene COL24A1. In another aspect, the polymorphism is in two genes, COL24A1 and SIGMAR1. In another aspect, the polymorphism is in three genes, APOE4, COL24A1, and SIGMAR1. In another aspect, the polymorphism is in two genes, APOE4 and COL24A1. In another aspect, the polymorphism is in four genes, APOE4, COL24A1, ADRA1 D, and15108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center ANXA2. In another aspect, the polymorphism is in one gene selected from APOE4, COL24A1, ADRA1 D, ANXA2, and SIGMAR1. In another aspect, the polymorphism is in one gene selected from APOE4, COL24A1, ADRA1 D, and ANXA2. In another aspect, the polymorphism is in APOE4. In another aspect, the polymorphism is in COL24A1. In another aspect, the polymorphism is in ADRA1D. In another aspect, the polymorphism is in ANXA2.

[0044] In some aspects, the polymorphism(s) associated with response to Sigma-1 receptor therapy (e.g., Sigma-1 receptor agonist therapy) is / are selected from the group consisting of APOE4 (rs429358 and / or rs7412), COL24A1 (rs60891279), ADRA1D (re184418609), ANXA2 (rs17845226), SIGMAR1_Q2P, COMT_L146fs, COMT_L146fs, KANSL1_P1010L / P304L / P946L, DHCR7_M220T, HLA-DRB1_ A244T, HLA-DRB1_ S66Y, HLA-DRB1_ Y89S, MS4A6E_M59T, RIN3_ H215R, DPYD_I543V, or combinations thereof. Table A details thesepolymorphisms.16108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center17108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center

[0045] In one aspect, the polymorphism associated with response to Sigma-1 receptor therapy is selected from the polymorphisms of Table A. In one aspect, the polymorphism associated with response to Sigma-1 receptor therapy is selected from the group consisting of APOE4 (rs429358 and / or rs7412), COL24A1 (rs60891279), ADRA1D (re184418609), ANXA2 (rs17845226), SIGMAR1_Q2P, COMT_L146fs (rs113895332 / rs61143203), KANSL1_P1010L / P304L / P946L, and combinations thereof. In another aspect, the polymorphism associated with response to Sigma-1 receptor therapy is selected from the group consisting of18108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center APOE4 (rs429358 and / or rs7412), COL24A1 (rs60891279), ADRA1D(re184418609), ANXA2 (rs17845226), SIGMAR 1_Q2P, COMT_L146fs(rs113895332 and / or rs61143203), KANSL1_P1010L / P304L / P946L, and combinations thereof. In yet another aspect, the polymorphism associated with response to Sigma-1 receptor therapy is COL24A1 (rs60891279). In yet another aspect, the polymorphism associated with response to Sigma-1 receptor therapy is ADRA1 D (re184418609). In yet another aspect, the polymorphism associated with response to Sigma-1 receptor therapy is ANXA2 (rs17845226). In yet another aspect, the polymorphism associated with response to Sigma-1 receptor therapy is SIGMAR1_Q2P. In one aspect, the polymorphism associated with response to Sigma-1 receptor therapy is COMT_L146fs (rs113895332 and / or rs61143203). In another aspect, the polymorphism associated with response to Sigma-1 receptor therapy is KANSL1_P1010L / P304L / P946L. In another aspect, the polymorphism associated with response to Sigma-1 receptor therapy is selected from the group consisting of APOE4 (rs429358 and / or rs7412), COL24A1 (rs60891279), ADRA1D (re184418609), ANXA2 (rs17845226), SIGMAR1_Q2P, COMT_L146fs(rs113895332 and / or rs61143203), and combinations thereof. In yet another aspect, the polymorphisms associated with response to Sigma-1 receptor therapy are COL24A1 (rs60891279) and SIGMAR1_Q2P. In yet another aspect, the polymorphisms associated with response to Sigma-1 receptor therapy are selected from the group consisting of APOE4 (rs429358 and / or rs7412), COL24A1 (rs60891279), ADRA1D (re184418609), ANXA2 (rs17845226), and combinations thereof. In yet another aspect, the polymorphisms associated with response to Sigma-1 receptor therapy are selected from the group consisting of COL24A1 (rs60891279), ADRA1D (re184418609), ANXA2 (rs17845226), and combinations thereof.

[0046] In various aspects, the Sigma-1 receptor therapy is Sigma-1 receptor agonist therapy.

[0047] It will be understood that any of the polymorphisms in Table A may be associated with response to Sigma-1 receptor therapy. For example, the polymorphism may be any one or more of the APOE4, COL24A1, ADRA1D, and19108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center ANXA2 polymorphisms detailed in Table A, or any other polymorphisms now known or identified later that can be associated with response to Sigma-1 receptor therapy.II. Method of Determining the Responsiveness of a Subject to Sigma-1 Receptor Therapy

[0048] Another aspect of the present invention provides methods of determining if a subject is responsive or non-responsive to Sigma-1 receptor therapy, such as Sigma-1 receptor agonist therapy. The method can comprise detecting the presence or absence of a gene associated with an increased or decreased response to Sigma-1 receptor therapy. The method can comprise detecting the presence of a polymorphism associated with an altered response (e.g., an increased or decreased response) to Sigma-1 receptor therapy. The method can also comprise determining the level of expression of RNA encoded by at least one gene associated with an altered response (e.g., an increased or decreased response) to Sigma-1 receptor therapy.

[0049] In one aspect, the method comprises detecting the presence of at least one gene associated with an increased response to Sigma-1 receptor therapy. The presence of the gene identifies a subject responsive to Sigma-1 receptor therapy. The absence of the gene identifies a subject non-responsive to Sigma-1 receptor therapy. The genes are as described in Section 1 above.

[0050] In one aspect, the method comprises detecting the presence of at least one polymorphism associated with an altered response to Sigma-1 receptor therapy. The presence of at least one polymorphism identifies a subject responsive to Sigma-1 receptor therapy. The absence of at least one polymorphism identifies a subject non-responsive to Sigma-1 receptor therapy. The polymorphisms and the genes comprising the polymorphisms are as described in Section I above.

[0051] The presence of one or more than one gene can be detected. For instance, the presence of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more genes can be detected. In some aspects, the presence of 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 genes can be detected. In other aspects, the presence of 1, 2, 3, 4, or 5 genes can be detected. In yet other aspects, the presence of 1, 2, or 3 genes can be detected. In yet other20108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center aspects, the presence of 1 or 2 genes can be detected. In yet other aspects, the presence of a single gene ( / .e., 1 gene) can be detected.

[0052] Additionally, or alternatively, the presence of one or more than one polymorphism can be detected. For instance, the presence of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more polymorphisms can be detected. In some aspects, the presence of 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 polymorphisms can be detected. In other aspects, the presence of 1, 2, 3, 4, or 5 polymorphisms can be detected. In yet other aspects, the presence of 1, 2, or 3 polymorphisms can be detected. In yet other aspects, the presence of 1 or 2 polymorphisms can be detected. In yet other aspects, the presence of a single polymorphism ( / .e., 1 polymorphism) can be detected.

[0053] The presence or absence of genes and / or polymorphisms can be detected by evaluating nucleic acid sequences in the subject. Alternatively, the presence or absence of genes and / or polymorphisms can be detected by evaluating the expression of a nucleic acid or protein from a gene or a genetic locus comprising the polymorphism.

[0054] In some aspects, the presence or absence of genes and / or polymorphisms can be detected by evaluating nucleic acid sequences for the presence of a gene or a polymorphism at the locus comprising the polymorphism. The nucleic acid sequence can be a DNA or an RNA sequence expressed from the gene or from a genetic locus comprising the polymorphism. Methods suitable for detecting genes and polymorphisms are well known in the art. For instance, the methods include shotgun sequencing, bridge PCR, high throughput methods, allelespecific real time PCR, 5'-nuclease assays, template-directed dye-terminator incorporation, molecular beacon allele-specific oligonucleotide assays, assays employing invasive cleavage with Flap nucleases, allele-specific hybridization (ASH), array based hybridization, allele-specific ligation, primer extension, single-base extension (SBE) assays, sequencing, pyrophosphate sequencing, real-time pyrophosphate sequencing, sequence length polymorphism analysis, restriction length fragment polymorphisms (RFLP), RFLP-PCR, single-stranded conformational polymorphism (SSCP), PCR-SSCP, fragment sizing capillary electrophoresis, heteroduplex analysis, and mass array systems. Analysis of amplified sequences may be performed using various technologies such as microchips, fluorescence21108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center polarization assays, and matrix-assisted laser desorption ionization (MALDI) mass spectrometry. High throughput sequencing methods include massively parallel signature sequencing (MPSS), polony sequencing, 454 pyrosequencing, illumina (Solexa) sequencing, combinatorial probe anchor synthesis (cPAS), SOLiD sequencing, ion Torrent semiconductor sequencing, DNA nanoball sequencing, heliscope single molecule sequencing, single molecule real time (SMRT) sequencing, nanopore DNA sequencing, tunneling currents DNA sequencing, sequencing by hybridization, sequencing with mass spectrometry, microfluidic Sanger sequencing, microscopy-based techniques, RNAP sequencing, and in vitro virus high-throughput sequencing.

[0055] In another aspect, the method comprises determining the level of expression of RNA encoded by at least one gene associated with an altered response to Sigma-1 receptor therapy, such as a Sigma-1 receptor agonist therapy. According to the method, a level of the RNA substantially similar to the level of the RNA in the subject responsive to Sigma-1 receptor therapy identifies a subject responsive to Sigma-1 receptor therapy. The gene(s) associated with an altered response to Sigma-1 receptor therapy are selected from the group consisting of APOE4, COL24A1, ADRA1D, ANXA2, SIGMAR1, COMT, KANSL1, DHCR7, HLA-DRB1, MS4A6E, RIN3, DPYD, and combinations thereof. In one aspect, the genes associated with an altered response to Sigma-1 receptor therapy are selected from the group consisting of APOE4, COL24A1, ADRA1D, ANXA2, SIGMAR1, COMT, KANSL1, and combinations thereof. In another aspect, the genes associated with an altered response to Sigma-1 receptor therapy are selected from the group consisting of APOE4, COL24A1, ADRA1 D, ANXA2, SIGMAR1, COMT, and combinations thereof. In yet another aspect, the genes associated with an altered response to Sigma-1 receptor therapy are selected from the group consisting of APOE4, COL24A1, ADRA1D, ANXA2, SIGMAR1, and combinations thereof. In yet another aspect, the genes associated with an altered response to Sigma-1 receptor therapy are selected from the group consisting of APOE4, COL24A1, ADRA1 D, ANXA2, and combinations thereof. In yet another aspect, the genes associated with an altered response to Sigma-1 receptor therapy are selected from the group consisting of COL24A1, ADRA1 D, ANXA2, and combinations thereof. In yet another aspect, the genes associated with an altered response to Sigma-1 receptor therapy 22108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center are APOE4, COL24A1 and / or SIGMAR1. In yet another aspect, the genes associated with an altered response to Sigma-1 receptor therapy are COL24A1 and SIGMAR1. In yet another aspect, the genes associated with an altered response to Sigma-1 receptor therapy are COL24A1, ADRA1 D, ANXA2, and SIGMAR1. In yet another aspect, the genes associated with an altered response to Sigma-1 receptor therapy are APOE4 and COL24A1. In yet another aspect, the genes associated with an altered response to Sigma-1 receptor therapy are selected from the group consisting of APOE4, COL24A1, ADRA1D, ANXA2, COMT, and combinations thereof. In yet another aspect, the genes associated with an altered response to Sigma-1 receptor therapy are APOE4 and SIGMAR1. In yet another aspect, the gene associated with an altered response to Sigma-1 receptor therapy is SIGMAR1. In yet another aspect, the gene associated with an altered response to Sigma-1 receptor therapy is APOE4. In yet another aspect, the gene associated with an altered response to Sigma-1 receptor therapy is COL24A1. In yet another aspect, the gene associated with an altered response to Sigma-1 receptor therapy is ADRA1 D. In yet another aspect, the gene associated with an altered response to Sigma-1 receptor therapy is ANXA2.

[0056] For instance, the level of expression of RNA encoded by 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more genes can be detected. In some aspects, the level of expression of RNA encoded by 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 genes can be detected. In other aspects, the level of expression of RNA encoded by 1, 2, 3, 4, or 5 genes can be detected. In yet other aspects, the level of expression of RNA encoded by of 1, 2, or 3 genes can be detected. In yet other aspects, the level of expression of RNA encoded by 1 or 2 genes can be detected. In yet other aspects, the level of expression of RNA encoded by a single gene ( / .e., 1 gene) can be detected.

[0057] Measuring the level of RNA expression may be accomplished by a variety of methods including northern blotting, quantitative real-time PCR (qRT-PCR), nucleic acid microarrays, Luminex microspheres, and nuclease protection assay. Methods of determining a substantially different level of RNA expression are known in the art, and include distribution analysis. In some aspects, a substantially different level of RNA expression is determined by normalizing RNA expression values as Transcripts Per Kilobase Million (TPM).23108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center

[0058] In other aspects, determining if a subject is responsive or non-responsive to Sigma-1 receptor therapy can be detected by measuring the level of a protein expressed from a gene. Methods of measuring protein expression include a variety of methods such as high-performance liquid chromatography (HPLC), liquid chromatography-mass spectrometry (LC / MS), enzyme-linked immunosorbent assay (ELISA), protein immunoprecipitation, immunoelectrophoresis, western blotting, and protein immunostaining.

[0059] In other aspects, determining if a subject is responsive or non-responsive to Sigma-1 receptor therapy can be detected by measuring the level of a protein expressed from a gene(s) comprising the polymorphism(s). Methods of measuring protein expression include a variety of methods such as high-performance liquid chromatography (HPLC), liquid chromatography-mass spectrometry (LC / MS), enzyme-linked immunosorbent assay (ELISA), protein immunoprecipitation, immunoelectrophoresis, western blotting, and protein immunostaining.

[0060] Determining if a subject is responsive or non-responsive to Sigma-1 receptor therapy can be detected in the subject in vivo or in vitro. Typically, responsiveness of a subject to Sigma-1 receptor therapy can be by analyzing a nucleic acid or protein in vitro in a biological sample obtained from the subject. The nucleic acid or protein can be isolated from the biological sample using methods commonly known in the art. For more information, see Ausubel et al., 2003, or Sambrook & Russell, 2001. Commercially available nucleic acid and protein extraction kits or commercially available extraction reagents may be used to isolate the nucleic acid from the biological sample.

[0061] Non-limiting examples of suitable biological samples include fluid samples, biopsy samples, skin samples, and hair samples. Fluid samples may include blood, serum, saliva, tears, and lymph. In some aspects the biological sample is blood. Methods of collecting a biological sample from a subject are well known in the art. Further disclosure regarding biological samples can be found in Section V below.

[0062] In some aspects the method of determining if a subject is responsive or non-responsive to Sigma-1 receptor therapy comprises obtaining the results of a test 24108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center of a biological sample from the subject which determines the presence of at least one gene and / or at least one polymorphism. In other aspects, the method of determining responsiveness of a subject to Sigma-1 receptor therapy comprises detecting if at least one polymorphism is present at a genetic locus in a biological sample obtained from the subject by sequencing the genetic locus comprising the polymorphism. In yet other aspects, the method of determining if a subject is responsive or non-responsive to Sigma-1 receptor therapy comprises obtaining the results of a test of a biological sample from the subject which determines the level of at least one RNA encoded by a gene associated with an altered response to Sigma-1 receptor therapy. In other aspects, the method of determining responsiveness of a subject to Sigma-1 receptor therapy comprises detecting if at least one polymorphism is present at a genetic locus in a biological sample obtained from the subject by determining the level of expression of at least one RNA encoded by a gene associated with an altered response to Sigma-1 receptor therapy.III. Methods of Using

[0063] The methods can be used to optimize treatment for subjects in need of Sigma-1 receptor therapy, such as Sigma-1 receptor agonist therapy. For instance, the methods can be used to select a subject responsive to Sigma-1 receptor therapy prior to initiating a treatment protocol for the subject. Conversely, the methods can be used to exclude, prior to treatment, a subject if the subject is found to be non-responsive to Sigma-1 receptor therapy. Polymorphisms can be used to determine, prior to treatment, if a subject will develop toxicity if treated with a Sigma-1 receptor agonist. Additionally, the methods can be used to determine if or when a treatment with a Sigma-1 receptor agonist should be discontinued in a subject. Additionally, the methods can be used in evaluating the response of a subject to a substance cessation treatment.

[0064] The methods comprise detecting the presence of a gene and / or a polymorphism in a subject, and determining if a subject is a candidate for Sigma-1 receptor therapy. The methods can also comprise determining the level of expression of RNA from one or more genes associated with an altered response to Sigma-1 receptor therapy. The methods can also comprise determining the level of 25108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center expression of RNA from one or more polymorphisms associated with an altered response to Sigma-1 receptor therapy. The polymorphisms and / or the genes associated with an altered response to Sigma-1 receptor therapy, the methods of detecting the genes and / or polymorphisms, and the methods of determining the level of RNA expression can be as described in Section I above, and methods of detecting the polymorphisms can be as described in Section II above.

[0065] As used herein, an “altered” response means the absence of a particular gene(s) and / or a polymorphism(s) results in a different response by the subject to a therapy than the response of a subject with the presence of a particular gene(s) and / or polymorphism(s), and vice versa. As used herein, the term “responsive” can be used to describe any subject that experiences a positive outcome when treated with a Sigma-1 receptor agonist. As used herein, the term “non-responsive” can be used to describe any subject that experiences a negative outcome when treated with a Sigma-1 receptor agonist. Methods of determining responsiveness of a subject to Sigma-1 receptor therapy can and will vary depending on the condition or disease, the subject, the treatment protocol, among other variables, and can be determined experimentally. Responsiveness can be as determined at any time point considered appropriate by an individual of skill in the art for a disease or condition. For instance, treatment outcome can be determined at about 57 weeks, about½8 weeks, or about 208 weeks after initiation of treatment.

[0066] When treatment is used for Alzheimer’s disease (AD), for example, positive outcomes can be as measured by Delta or Slope MMSE (Mini-Mental State Examination) or ADCS-ADL (Alzheimer's Disease Cooperative Study - Activities of Daily Living). Responsive subjects are characterized by having a positive or null MMSE and / or ADCS-ASL slope, whereas non-responsive subjects are characterized by having a negative MMSE and / or ADCS-ASL slope. Responsive subjects are also characterized by having a high MMSE and / or ADCS-ASL delta, whereas non-responsive subjects are characterized by having a low MMSE and / or ADCS-ASL delta. A low MMSE delta may range from about -9 to about -6, from about -6 to about -2, or from about -9 to about -2, whereas a high MMSE delta may range from about -6 to -2 or from about -2 to about 6. Methods of determining Delta or Slope MMSE or ADCS-ADL can be as described in the examples herein.26108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center

[0067] Positive outcomes of Alzheimer’s disease treatment can additionally or alternatively be measured, for instance, by Alzheimer's Disease Assessment Scale-Cognitive (ADAS-Cog13), Clinical Dementia Rating-Sum of Boxes (CDR-SB), Clinical Global Impressions Scale (CGI) - Improvement (CGI-I), and / or Neuropsychiatric Inventory Questionnaire (NPI-Q).

[0068] In some aspects, the method is used to select a subject responsive to Sigma-1 receptor therapy, such as Sigma-1 receptor agonist therapy. The method comprises obtaining the results of a test of a biological sample from the subject which determines the presence of at least one gene and / or at least one polymorphism, and excluding the subject as non-responsive to Sigma-1 receptor agonist therapy if a gene and / or a polymorphism is present in the subject.Alternatively, the method comprises obtaining the results of a test of a biological sample from the subject which determines the presence of at least one gene and / or at least one polymorphism, and including the subject as responsive to Sigma-1 receptor agonist therapy if a gene and / or a polymorphism is present in the subject.

[0069] In one aspect, the gene is selected from the group consisting of APOE4, COL24A1, ADRA1D, ANXA2, SIGMAR1, COMT, KANSL1, DHCR7, HLA-DRB1, MS4A6E, RIN3, DPYD, and combinations thereof. In one aspect, the polymorphism is selected from the group consisting of APOE4 (rs429358 and / or rs7412), COL24A1 (rs60891279), ADRA1D (re184418609), ANXA2 (rs17845226), SIGMAR1_Q2P, COMT_L146fs, KANSL1_P1010L / P304L / P946L, DHCR7_M220T, HLA-DRB1_ A244T, HLA-DRB1_ S66Y, HLA-DRB1_ Y89S, MS4A6E_M59T, RIN3_ H215R, DPYD_I543V, and combinations thereof. In another aspect, the polymorphism is selected from the group consisting of APOE4 (rs429358 and / or rs7412), COL24A1 (rs60891279), ADRA1D (re184418609), ANXA2 (rs17845226), SIGMAR1_Q2P, COMT_L146fs, KANSL1_P1010L / P304L / P946L, and combinations thereof. In another aspect, the polymorphism is selected from the group consisting of APOE4 (rs429358 and / or rs7412), COL24A1 (rs60891279), ADRA1D(re184418609), ANXA2 (rs17845226), SIGMAR1_Q2P, and combinations thereof. In another aspect, the polymorphism is selected from the group consisting of APOE4 (rs429358 and / or rs7412), COL24A1 (rs60891279), ADRA1D (re184418609),27108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center ANXA2 (rs17845226), and combinations thereof. The presence of the gene and / or the polymorphism can be obtained by evaluating nucleic acid sequences.

[0070] In other aspects, the method is used to select a subject responsive to Sigma-1 receptor therapy (e.g., Sigma-1 receptor agonist therapy). The method comprises detecting if at least one polymorphism is present at a genetic locus in a biological sample obtained from the subject by sequencing the genetic locus comprising the polymorphism, and excluding the subject as non-responsive to Sigma-1 receptor agonist therapy if a polymorphism is present in the subject.Alternatively, a subject can be included as responsive to the therapy if a polymorphism is present in the subject. In one aspect, the polymorphism is selected from the group consisting of APOE4 (rs429358 and / or rs7412), COL24A1 (rs60891279), ADRA1D (re184418609), ANXA2 (rs17845226), SIGMAR1_Q2P, COMT_L146fs, KANSL1_P1010L / P304L / P946L, DHCR7_M220T, HLA-DRB1_ A244T, HLA-DRB1_ S66Y, HLA-DRB1_ Y89S, MS4A6E_M59T, RIN3_ H215R, DPYD_I543V, and combinations thereof. In another aspect, the polymorphism is selected from the group consisting of APOE4 (rs429358 and / or rs7412), COL24A1 (rs60891279), ADRA1D (re184418609), ANXA2 (rs17845226), SIGMAR1_Q2P, COMT_L146fs, and combinations thereof. In another aspect, the polymorphism is selected from the group consisting of APOE4 (rs429358 and / or rs7412), COL24A1 (rs60891279), ADRA1D (re184418609), ANXA2 (rs17845226), SIGMAR1_Q2P, and combinations thereof. In another aspect, the polymorphism is selected from the group consisting of APOE4 (rs429358 and / or rs7412), COL24A1 (rs60891279), ADRA1 D (re184418609), ANXA2 (rs17845226), and combinations thereof. In another aspect, the polymorphism is selected from the group consisting of COL24A1 (rs60891279), ADRA1 D (re184418609), ANXA2 (rs17845226), and combinations thereof. The presence of the polymorphism can be detected by evaluating nucleic acid sequences.

[0071] In other aspects, the method is used to select a subject responsive to Sigma-1 receptor therapy, such as Sigma-1 receptor agonist therapy. The method comprises obtaining the results of a test of a biological sample from the subject which determines the level of expression of RNA encoded by at least one gene selected from the group consisting of APOE4, COL24A1, ADRA1 D, ANXA2,28108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center SIGMAR1, COMT, KANSL1, DHCR7, HLA-DRB1, MS4A6E, RIN3, and combinations thereof, and excluding the subject as non-responsive to Sigma-1 receptor agonist therapy if a level of RNA is substantially different from the level of RNA in a subject responsive to Sigma-1 receptor therapy. Alternatively, a subject can be included as responsive to the therapy if a level of RNA is substantially similar to the level of the RNA in a subject responsive to Sigma-1 receptor therapy. In one aspect, the level of expression of RNA encoded by a gene selected from the group consisting of APOE4, COL24A1, ADRA1 D, ANXA2, SIGMAR1, COMT, KANSL1, and combinations thereof, is determined. In one aspect, the level of expression of RNA encoded by a gene selected from the group consisting of APOE4, COL24A1, SIGMAR1, ADRA1D, ANXA2, COMT, and combinations thereof, is determined. In one aspect, the level of expression of RNA encoded by a gene selected from the group consisting of APOE4, COL24A1, ADRA1D, ANXA2, SIGMAR1, and combinations thereof, is determined. In one aspect, the level of expression of RNA encoded by a gene selected from the group consisting of APOE4, COL24A1, ADRA1 D, ANXA2, and combinations thereof, is determined. In one aspect, the level of expression of RNA encoded by a gene selected from the group consisting of COL24A1, ADRA1D, ANXA2, and combinations thereof, is determined.

[0072] In some aspects, the method is used to select subjects responsive to Sigma-1 receptor agonist therapy (e.g., Sigma-1 receptor agonist therapy). The method comprises detecting RNA expression in a test of a biological sample from the subject which determines the level of expression of RNA encoded by at least one gene selected from the group consisting of APOE4, COL24A1, ADRA1D, ANXA2, SIGMAR1, COMT, KANSL1, DHCR7, HLA-DRB1, MS4A6E, RIN3, and combinations thereof. The method comprises excluding the subject as non-responsive to Sigma-1 receptor agonist therapy if a level of RNA is substantially different from the level of RNA in a subject responsive to Sigma-1 receptor therapy. Alternatively, the method comprises identifying the subject as responsive to Sigma-1 receptor agonist therapy if a level of RNA is substantially similar to the level of RNA in a subject responsive to Sigma-1 receptor therapy. In one aspect, the level of expression of RNA encoded by a gene selected from the group consisting of APOE4, COL24A1, ADRA1D, ANXA2, SIGMAR1, COMT, KANSL1, and combinations thereof, is determined. In one aspect, the level of expression of RNA 29108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center encoded by a gene selected from the group consisting of APOE4, COL24A1, ADRA1D, ANXA2, SIGMAR1, COMT, and combinations thereof, is determined. In one aspect, the level of expression of RNA encoded by a gene selected from the group consisting of APOE4, COL24A1, ADRA1D, ANXA2, SIGMAR1, and combinations thereof, is determined. In another aspect, the level of expression of RNA encoded by a gene selected from APOE4, COL24A1 and SIGMAR1, is determined. In another aspect, the level of expression of RNA encoded by a gene selected from APOE4, COL24A1, ADRA1 D, and ANXA2 is determined. In another aspect, the level of expression of RNA encoded by a gene selected from COL24A1, ADRA1 D, and ANXA2 is determined.

[0073] In some aspects, the method is used to treat a subject in need of Sigma-1 receptor therapy, such as Sigma-1 receptor agonist therapy. The method comprises obtaining the results of a test of a biological sample from the subject which determines the presence of at least one polymorphism. If a polymorphism is not present in the subject, a therapeutically effective amount of a Sigma-1 receptor agonist is administered to the subject. Alternatively, if a polymorphism is present in the subject, a therapeutically effective amount of an alternative to Sigma-1 receptor agonist is administered to the subject. In one aspect, the polymorphism is selected from the group consisting of APOE4 (rs429358 and / or rs7412), COL24A1 (rs60891279), ADRA1D (re184418609), ANXA2 (rs17845226), SIGMAR1_Q2P, COMT_L146fs, KANSL1_P1010L / P304L / P946L, DHCR7_M220T, HLA-DRB1_ A244T, HLA-DRB1_ S66Y, HLA-DRB1_ Y89S, MS4A6E_M59T, RIN3_ H215R, DPYD_I543V, and combinations thereof. In another aspect, the polymorphism is selected from the group consisting of APOE4 (rs429358 and / or rs7412), COL24A1 (rs60891279), ADRA1D (re184418609), ANXA2 (rs17845226), SIGMAR1_Q2P, COMT_L146fs, COMT_L146fs, KANSL1_P1010L / P304L / P946L, and combinations thereof. In another aspect, the polymorphism is selected from the group consisting of APOE4 (rs429358 and / or rs7412), COL24A1 (rs60891279), ADRA1D(re184418609), ANXA2 (rs17845226), SIGMAR1_Q2P, COMT_L146fs, COMT_L146fs, and combinations thereof. In another aspect, the polymorphism is selected from the group consisting of APOE4 (rs429358 and / or rs7412), COL24A1 (rs60891279), ADRA1D (re184418609), ANXA2 (rs17845226), SIGMAR1_Q2P, and combinations thereof. In another aspect, the polymorphism is selected from the 30108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center group consisting of APOE4 (rs429358 and / or rs7412), COL24A1 (rs60891279), ADRA1 D (re184418609), ANXA2 (rs17845226), and combinations thereof. In another aspect, the polymorphism is selected from the group consisting of COL24A1 (rs60891279), ADRA1 D (re184418609), ANXA2 (rs17845226), and combinations thereof. In another aspect, the polymorphisms are APOE4 (rs429358 and / or rs7412), COL24A1 (rs60891279) and SIGMAR1_Q2P. In another aspect, the polymorphism is APOE4 (rs429358). In another aspect, the polymorphism is COL24A1 (rs60891279). In another aspect, the polymorphism is ADRA1D(re184418609). In another aspect, the polymorphism is ANXA2 (rs17845226). In another aspect, the polymorphism is SIGMAR1_Q2P. The presence of the polymorphism can be obtained by evaluating nucleic acid sequences.

[0074] In other aspects, the method is used to treat a subject in need of Sigma-1 receptor therapy (e.g., Sigma-1 receptor agonist therapy). The method comprises obtaining the results of a test of a biological sample from the subject which determines the level of expression of RNA encoded by a gene selected from the group consisting of APOE4, COL24A1, ADRA1D, ANXA2, SIGMAR1, COMT, KANSL1, DHCR7, HLA-DRB1, MS4A6E, RIN3, and combinations thereof. If the level of RNA in the test sample is substantially similar to the level of RNA obtained from a subject responsive to Sigma-1 receptor therapy, a therapeutically effective amount of a Sigma-1 receptor agonist is administered to the subject. Alternatively, if the level of RNA in the test sample is substantially different from the level of RNA obtained from a subject responsive to Sigma-1 receptor therapy, a therapeutically effective amount of an alternative to Sigma-1 receptor agonist is administered to the subject. In another aspect, the level of expression of RNA encoded by a gene selected from the group consisting of APOE4, COL24A1, ADRA1 D, ANXA2, SIGMAR1, COMT, KANSL1, and combinations thereof, is determined. In another aspect, the level of expression of RNA encoded by a gene selected from the group consisting of APOE4, COL24A1, ADRA1 D, ANXA2, SIGMAR1, COMT, and combinations thereof, is determined. In another aspect, the level of expression of RNA encoded by a gene selected from the group consisting of APOE4, COL24A1, SIGMAR1, and combinations thereof, is determined. In another aspect, the level of expression of RNA encoded by a gene selected from the group consisting of APOE4, COL24A1, ADRA1D, ANXA2, and combinations thereof, is determined. In 31108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center another aspect, the level of expression of RNA encoded by a gene selected from the group consisting of COL24A1, ADRA1 D, ANXA2, and combinations thereof, is determined.

[0075] It will be recognized that a subject can have more than one genes relevant to responsiveness or non-responsiveness to Sigma-1 receptor therapy. For instance, the subject can have 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more genes relevant to responsiveness or non-responsiveness to Sigma-1 receptor therapy. In some aspects, the subject has 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 genes relevant to responsiveness or non-responsiveness to Sigma-1 receptor therapy. In other aspects, the subject has 1, 2, 3, 4, or 5 genes relevant to responsiveness or non-responsiveness to Sigma-1 receptor therapy. In yet other aspects, the subject has 1, 2, or 3 genes relevant to responsiveness or non-responsiveness to Sigma-1 receptor therapy. In yet other aspects, the subject has 1 or 2 genes relevant to responsiveness or non-responsiveness to Sigma-1 receptor therapy. In yet other aspects, the subject has a single gene ( / .e., 1 gene) relevant to responsiveness or non-responsiveness to Sigma-1 receptor therapy.

[0076] It will be recognized that a subject can have more than one polymorphism relevant to responsiveness or non-responsiveness to Sigma-1 receptor therapy. For instance, the subject can have 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more polymorphisms relevant to responsiveness or non-responsiveness to Sigma-1 receptor therapy. In some aspects, the subject has 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 polymorphisms relevant to responsiveness or non-responsiveness to Sigma-1 receptor therapy. In other aspects, the subject has 1, 2, 3, 4, or 5 polymorphisms relevant to responsiveness or non-responsiveness to Sigma-1 receptor therapy. In yet other aspects, the subject has 1, 2, or 3 polymorphisms relevant to responsiveness or non-responsiveness to Sigma-1 receptor therapy. In yet other aspects, the subject has 1 or 2 polymorphisms relevant to responsiveness or non-responsiveness to Sigma-1 receptor therapy. In yet other aspects, the subject has a single polymorphism ( / .e., 1 polymorphism) relevant to responsiveness or non-responsiveness to Sigma-1 receptor therapy.32108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center IV. Treating a Subject in Need of Sigma-1 Receptor Therapy

[0077] Sigma-1 receptor expression or activity is linked to neurodegeneration, and the activation of Sigma-1 receptor is associated with neuroprotection in human subjects and in different in vitro and in vivo models. Therefore, one aspect of the disclosure encompasses treatment of any Sigma-1 receptor-related disease or condition which primarily affects the neuronal system in a subject. Such conditions are variously known as neurodevelopmental and neurodegenerative diseases, disorders, and conditions, and can additionally or alternatively be used for neuroprotection. Accordingly, a subject in need of Sigma-1 receptor therapy has a condition selected from the group consisting of a neuroprotective condition, a neurodegenerative condition, and a neurodevelopmental condition.

[0078] Non-limiting examples of neuroprotective condition, a neurodegenerative condition, and a neurodevelopmental condition include cognitive impairment, brain atrophy, Lewy body dementia, stroke, traumatic brain injury, infections, spinal cord injury, Alzheimer’s disease (AD) (including early onset AD (EOAD), discussed below), Parkinson’s disease, dementia, Huntington’s disease, Amyotrophic lateral sclerosis, Prion disease, Rett Syndrome, Fragile X Syndrome, cerebral palsy, Angelman syndrome, Williams syndrome, pervasive developmental disorder not otherwise specified (PDD-NOS), childhood disintegrative disorder, Smith-Magenis syndrome, multiple sclerosis, Fronto-temporal dementia, motor neuron disease (MND), spinocerebellar ataxia (SCA), spinal muscular atrophy (SMA), autism spectrum disorder, schizophrenia, post-traumatic stress disorder (PTSD), and any combination thereof

[0079] Certain genes, such as SIGMAR1, are also useful as a target for cancer treatment. As such, one aspect of the disclosure encompasses treatment of any Sigma-1 receptor-related cancer or neoplasm. As it will be recognized by individuals skilled in the art, cancer as used throughout the instant disclosure may be one or more neoplasm or cancer. The neoplasm may be malignant or benign, the cancer may be primary or metastatic; the neoplasm or cancer may be early stage or late stage. Non-limiting examples of neoplasms or cancers that may be treated include acute lymphoblastic leukemia, acute myeloid leukemia, adrenocortical carcinoma, AIDS-related cancers, AIDS-related lymphoma, anal cancer, appendix33108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center cancer, astrocytomas (childhood cerebellar or cerebral), basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer, brainstem glioma, brain tumors (cerebellar astrocytoma, cerebral astrocytoma / malignant glioma, ependymoma, medulloblastoma, supratentorial primitive neuroectodermal tumors, visual pathway and hypothalamic gliomas), breast cancer, bronchial adenomas / carcinoids, Burkitt lymphoma, carcinoid tumors (childhood, gastrointestinal), carcinoma of unknown primary, central nervous system lymphoma (primary), cerebellar astrocytoma, cerebral astrocytoma / malignant glioma, cervical cancer, childhood cancers, chronic lymphocytic leukemia, chronic myelogenous leukemia, chronic myeloproliferative disorders, colon cancer, cutaneous T-cell lymphoma, desmoplastic small round cell tumor, endometrial cancer, ependymoma, esophageal cancer, Ewing's sarcoma in the Ewing family of tumors, extracranial germ cell tumor (childhood), extragonadal germ cell tumor, extrahepatic bile duct cancer, eye cancers (intraocular melanoma, retinoblastoma), gallbladder cancer, gastric (stomach) cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor, germ cell tumors (childhood extracranial, extragonadal, ovarian), gestational trophoblastic tumor, gliomas (adult, childhood brain stem, childhood cerebral astrocytoma, childhood visual pathway and hypothalamic), gastric carcinoid, hairy cell leukemia, head and neck cancer, hepatocellular (liver) cancer, Hodgkin lymphoma, hypopharyngeal cancer, hypothalamic and visual pathway glioma (childhood), intraocular melanoma, islet cell carcinoma, Kaposi sarcoma, kidney cancer (renal cell cancer), laryngeal cancer, leukemias (acute lymphoblastic, acute myeloid, chronic lymphocytic, chronic myelogenous, hairy cell), lip and oral cavity cancer, liver cancer (primary), lung cancers (non-small cell, small cell), lymphomas (AIDS-related, Burkitt, cutaneous T-cell, Hodgkin, non-Hodgkin, primary central nervous system), macroglobulinemia (Waldenstrom), malignant fibrous histiocytoma of bone / osteosarcoma, medulloblastoma (childhood), melanoma, intraocular melanoma, Merkel cell carcinoma, mesotheliomas (adult malignant, childhood), metastatic squamous neck cancer with occult primary, mouth cancer, multiple endocrine neoplasia syndrome (childhood), multiple myeloma / plasma cell neoplasm, mycosis fungoides, myelodysplastic syndromes, myelodysplastic / myeloproliferative diseases, myelogenous leukemia (chronic), myeloid leukemias (adult acute, childhood acute), multiple myeloma, myeloproliferative disorders (chronic), nasal cavity and paranasal34108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center sinus cancer, nasopharyngeal carcinoma, neuroblastoma, non-Hodgkin lymphoma, non-small cell lung cancer, oral cancer, oropharyngeal cancer, osteosarcoma / malignant fibrous histiocytoma of bone, ovarian cancer, ovarian epithelial cancer (surface epithelial-stromal tumor), ovarian germ cell tumor, ovarian low malignant potential tumor, pancreatic cancer, pancreatic cancer (islet cell), paranasal sinus and nasal cavity cancer, parathyroid cancer, penile cancer, pharyngeal cancer, pheochromocytoma, pineal astrocytoma, pineal germinoma, pineoblastoma and supratentorial primitive neuroectodermal tumors (childhood), pituitary adenoma, plasma cell neoplasia, pleuropulmonary blastoma, primary central nervous system lymphoma, prostate cancer, rectal cancer, renal cell carcinoma (kidney cancer), renal pelvis and ureter transitional cell cancer, retinoblastoma, rhabdomyosarcoma (childhood), salivary gland cancer, sarcoma (Ewing family of tumors, Kaposi, soft tissue, uterine), Sezary syndrome, skin cancers (nonmelanoma, melanoma), skin carcinoma (Merkel cell), small cell lung cancer, small intestine cancer, soft tissue sarcoma, squamous cell carcinoma, squamous neck cancer with occult primary (metastatic), stomach cancer, supratentorial primitive neuroectodermal tumor (childhood), T-Cell lymphoma (cutaneous), testicular cancer, throat cancer, thymoma (childhood), thymoma and thymic carcinoma, thyroid cancer, thyroid cancer (childhood), transitional cell cancer of the renal pelvis and ureter, trophoblastic tumor (gestational), unknown primary site (adult, childhood), ureter and renal pelvis transitional cell cancer, urethral cancer, uterine cancer (endometrial), uterine sarcoma, vaginal cancer, visual pathway and hypothalamic glioma (childhood), vulvar cancer, Waldenstrom macroglobulinemia, and Wilms tumor (childhood).

[0080] In a particular aspect, the neuroprotective condition, neurodegenerative condition, and / or neurodevelopmental condition treated in accordance with the methods disclosed herein are associated with or related to Alzheimer’s disease (AD). Stages of AD include early signs of AD, early-onset AD, or late-onset Alzheimer. Early-onset Alzheimer’s disease (EOAD) refers to AD onset at an age of 65 years old or younger. EOAD is often delayed in diagnosis and overshadowed by the more common late-onset AD (LOAD). EOAD comprises about 5% of AD and differs significantly from LOAD, such as aggressive in its course and demanding age-related psychosocial support. A substantial percentage of EOAD is phenotypic 35108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center variants that differ from the usual memory-disordered presentation of typical AD. Patients with EOAD often have greater parietal atrophy, more white matter abnormalities, and less hippocampal volume loss, compared to those with LOAD. The phenotypic variants also have atrophy and white matter changes corresponding anatomically to the cognitive changes and appear to involve alternate neural networks relative to typical AD. Management of EOAD is similar to that for LOAD, but special emphasis should be placed on targeting the specific cognitive areas involved and more age-appropriate psychosocial support and education. In one aspect, the present disclosure provides therapy for both EOAD and LOAD. In another aspect, the present disclosure provides Sigma-1 receptor (e.g., agonist) therapy specifically for EOAD or early signs or early stages of AD ( / .e., mild AD). In one embodiment, the Alzheimer’s disease is early onset Alzheimer’s disease. In another embodiment, the Alzheimer’s disease is early stage or mild Alzheimer’s disease.

[0081] The method comprises administering a therapeutically effective amount of an alternative to Sigma-1 receptor agonist when a subject in need of Sigma-1 receptor agonist therapy is determined to be non-responsive to the therapy. Nonlimiting examples of alternatives to Sigma-1 receptor agonist include galantamine, memantine, and rivastigmine.

[0082] The method comprises administering a therapeutically effective amount of a Sigma-1 receptor agonist when a subject in need of Sigma-1 receptor agonist therapy is determined to be responsive to the therapy. A Sigma-1 receptor agonist may be any therapeutic or active agent capable of treating a subject in need of Sigma-1 receptor agonist therapy. For instance, a receptor agonist can be a biological compound such as an antibody or a peptide, ora small molecule active agent. Non-limiting examples of Sigma-1 receptor agonists include entacapone, nebicapone, nitecapone, opicapone, tolcapone, tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride (ANAVEX2-73, AV2-73, or A2-73), 1-(2,2-diphenyltetrahydrofuran-3-yl)-N-methylmethanamine hydrochloride (ANAVEX19-144, orA19-144), aminotetrahydrofuran derivative tetrahydro-N, N-dimethyl-5,5-diphenyl-3-furanmethanamine hydrochloride (ANAVEX1-41), 1-(3-4(((1R,3S,5S)-adamantan-1-yl)(phenyl)methyl)propyl)-4-methylpiperazine36108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center (ANAVEX™ 1066 or “AV1066”), ANAVEX3-71 (also known asAF710B), 1-[(2R)-2,8-dimethyl-1-thia-3,8-diazaspiro[4.5]decan-3-yl]-3-(1 H-indol-3-yl)propan-1-one (AF710A), 1-[2,8-dimethyl-1-thia-3,8-diazaspiro[4.5]decan-3-yl]-3-(1 H-indol-3-yl)propan-1-one (AF710), PRE-084, donepezil, fluvoxamine, amitriptyline, L-687,384, SA-4503, Dextromethorphan, Dimethyltryptamine, (+)-pentazocine, or any of their crystal forms, enantiomers and pharmaceutically acceptable salts thereof and combinations thereof. Sigma-1 receptor agonists may also be antibodies.

[0083] As used herein, ANAVEX® 2-73 (A2-73 or blarcamesine) has the chemical name of tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride. A2-73 freebase has the chemical name of tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine. ANAVEX®19-144 (A19-144) has the chemical name of 1-(2,2-diphenyltetrahydrofuran-3-yl)-N-methylmethanamine hydrochloride. ANAVEX 19-144 Freebase (A19-144 freebase) has the chemical name of 1 -(2,2-diphenyltetrahydrofuran-3-yl)-N-methylmethanamine. ANAVEX®1-41 (A1-41) has the chemical name of tetrahydro-N, N-dimethyl-5,5-diphenyl-3-furanmethanamine hydrochloride. ANAVEX 1-41 freebase (A1-41 freebase) has the chemical name of tetrahydro-N, N-dimethyl-5,5-diphenyl-3-furanmethanamine. AV1066 has the chemical name of 1-(3-4(((1R,3S,5S)-adamantan-1-yl) (pheny)methyl) propyl)-4-methylpiperazine. ANAVEX®3-71 (A3-71, AF-710B) has a chemical name of 1 -(2,8-dimethyl-1-thia-3,8-diazaspiro [4.5] decan-3-yl)-3-(1 H-indol-3-yl) propan-1-one. In one aspect, the Sigma-1 agonist comprises tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine (A2-73 freebase), a polymorph thereof, an enantiomer thereof, a polymorph of the enantiomer thereof, a pharmaceutically acceptable salt thereof, an enantiomer of the pharmaceutically acceptable salt thereof, a polymorph of the pharmaceutically acceptable salt thereof, or a co-crystal thereof. In another aspect, the agent comprises tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride (A2-73), a polymorph thereof, an enantiomer thereof, a polymorph of the enantiomer thereof, or a co-crystal thereof.

[0084] In various aspects of the methods disclosed in the present disclosure, the Sigma-1 receptor agonist comprises ANAVEX® 2-73 (A2-73) or A2-73 freebase, both of which are Sigma-1 receptor agonists which bind to the SIGMAR1 receptor on the cell membrane. A2-73 has been evaluated for therapeutic clinical efficacy in37108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center various neurodegenerative and / or neurodevelopmental disorders, such as Alzheimer’s Disease, Parkinson’s Disease, Rett syndrome, Angelman syndrome, Williams syndrome, Fragile X Syndrome, among others. A2-73 and the related compounds have demonstrated encouraging therapeutic efficacy for patients with these neurological diseases.

[0085] In various aspects, the Sigma-1 receptor agonist comprises tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine in amorphous form (A2-73 freebase amorphous form); tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine in crystal Form I (A2-73 freebase Crystal Form I); tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride in crystal Form I (A2-73 Crystal Form I); tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride in crystal Form II (A2-73 Crystal Form II); tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride in crystal Form III (A2-73 Crystal Form III); (+) tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride in amorphous form ((+) A2-73 amorphous form); (-) tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride in amorphous form ((-) A2-73 amorphous form); (+) tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride in a crystal form ((+) A2-73 crystal form); (-) tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride in a crystal form ((-) A2-73 crystal form); tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrogen fumarate in amorphous form (A2-73 hydrogen fumarate amorphous form); tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrogen fumarate in crystal Form I (A2-73 hydrogen fumarate Crystal Form I); tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrogen fumarate in crystal Form II (A2-73 hydrogen fumarate Crystal Form II); tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrogen fumarate in crystal Form III (A2-73 hydrogen fumarate Crystal Form III); tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrogen fumarate in crystal Form IV (A2-73 hydrogen fumarate Crystal Form IV); tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrogen fumarate in crystal Form V (A2-73 hydrogen fumarate Crystal Form V); tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine mesylate in crystal Form I (A2-73 mesylate Crystal Form I); tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine sulfate in crystal Form I (A2-73 sulfate Crystal Form I); tetrahydro-N, N-dimethyl-2,2-diphenyl-3- 38108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center furanmethanamine sulfate in crystal Form II (A2-73 sulfate Crystal Form II); tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine oxalate in crystal Form I (A2-73 oxalate Crystal Form I); tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine oxalate in crystal Form II (A2-73 oxalate Crystal Form II); tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine dihydrogen phosphate in crystal Form I (A2-73 dihydrogen phosphate Crystal Form I); tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine edisylate in crystal Form I (A2-73 edisylate Crystal Form I); tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine benzoate in crystal Form I (A2-73 benzoate Crystal Form I); tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrobromide in crystal Form A (A2-73 hydrobromide Crystal Form A); tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrobromide in crystal Form B (A2-73 hydrobromide Crystal Form B); tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine maleate in crystal Form S5 (A2-73 maleate Crystal Form S5); tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine maleate in crystal Form S6 (A2-73 maleate Crystal Form S6); co-crystal Form CSII of tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride with tartaric acid (A2-73 co-crystal with tartaric acid Crystal Form CSII); co-crystal Form CSIII of tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride with citric acid (A2-73 co-crystal with citric acid Crystal Form CSIII); co-crystal Form CSIV of tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride with malic acid (A2-73 co-crystal with malic acid Crystal Form CSIV); a co-crystal of tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride with zinc chloride (A2-73 co-crystal with zinc chloride); a co-crystal of (-) tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride with zinc chloride ((-) A2-73 co-crystal with zinc chloride); a co-crystal of (+) tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride with zinc chloride ((+) A2-73 co-crystal with zinc chloride); or any combination thereof.

[0086] In one aspect, the Sigma-1 receptor agonist comprises A2-73 freebase in amorphous form, A2-73 freebase in crystal Form I, A2-73 in amorphous form, A2-73 crystal Form I, A2-73 crystal Form II, A2-73 crystal Form III, (+)A2-73 enantiomer, (-) A2-73 enantiomer. The A2-73 freebase in crystal Form I is disclosed and characterized in WO 2019 / 200345. Specifically, the A2-73 Freebase in crystal Form I 39108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center is characterized substantially by XRPD pattern shown in FIG. 16 ofWO 2019 / 200345 and particle shapes depicted in FIG. 15 of WO 2019 / 200345. In one aspect, A2-73 crystal is the Form I, Form II, and / or Form III crystals as disclosed and characterized in WO 2017 / 013498. Specifically, the A2-73 crystal Form I is characterized substantially by XRPD pattern shown in FIG. 1 of WO 2017 / 013498 and particle shapes depicted in FIGs. 2 and 3 of WO 2017 / 013498. The A2-73 crystal Form II is characterized substantially by XRPD pattern shown in FIG. 4, particle shapes depicted in FIG. 7 of WO 2017 / 013498, and the FTIR spectrum shown in FIG. 5 of WO 2017 / 013498. The A2-73 crystal Form III is characterized substantially by XRPD pattern shown in FIG. 8, particle shapes depicted in FIG. 11 of WO 2017 / 013498, and the FTIR spectrum shown in FIG. 9 of WO 2017 / 013498. The analog of A2-73 may include A1 -41. The metabolite of A2-73 may include A19-144. In one aspect, the pharmaceutically acceptable salt comprises a hydrochloride salt, a hydrobromide salt, a fumarate salt, a sulfate salt, a dihydrogen phosphate salt, a benzoate salt, a mesylate salt, an edysilate salt, or an oxalate salt. In another aspect, the co-crystal comprises A2-73 pharmaceutically acceptable salt with an organic acid comprising tartaric acid, citric acid, maleic acid, or a combination thereof. In another aspect, the A2-73 is A2-73 amorphous form, A2-73 Freebase crystal Form I, A2-73 crystal Form I, A2-73 crystal Form II, A2-73 crystal Form III, (+)A2-73 enantiomer, (-)A2-73 enantiomer. In one aspect, the active is A2-73 freebase fumarate salt and / or its crystals, such as A2-73 Freebase fumarate salt in amorphous form, A2-73 Freebase hydrogen fumarate salt crystal Form I, A2-73 Freebase hydrogen fumarate salt crystal Form II, A2-73 Freebase hydrogen fumarate salt crystal Form III, A2-73 Freebase hydrogen fumarate salt crystal Form IV, A2-73 Freebase hydrogen fumarate salt crystal Form V, all of which are disclosed and characterized in WO 2019 / 200345, such as XRPD patterns shown in FIG. 29, FIG. 30, FIG. 32 FIG. 33, and FIG. 34, respectively. In another aspect, the A2-73 is other salt crystals, such as A2-73 Freebase Mesylate Form I, A2-73 Freebase Sulfate Form I, A2-73 Freebase Sulfate Form II, A2-73 Freebase Oxalate Form I, A2-73 Freebase Oxalate Form II, A2-73 Freebase Oxalate Form III, A2-73 Freebase Dihydrogen phosphate Form I, A2-73 Freebase Edisylate Form I, A2-73 Freebase Benzoate Form I, all of which are disclosed and characterized in WO 2019 / 200345, such as XRPD patterns shown in FIGs. 18-23 and 25-27, respectively. In yet another40108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center aspect, the active is A2-73 Freebase hydrobromide crystal Form A, A2-73 Freebase hydrobromide crystal Form B, A2-73 maleate crystal Form S5, or A2-73 maleate crystal Form S6. They are disclosed and characterized in WO 2021 / 158586. In yet another aspect, the active is co-crystal Form OSH of A2-73 co-crystal with tartaric acid, co-crystal Form CSIII of A2-73 co-crystal with citric acid, or co-crystal Form CSIV of A2-73 co-crystal with malic acid, which are disclosed and characterized in WO 2023 / 208133. In yet another aspect, the active is co-crystal of A2-73 with zinc chloride disclosed in U. S. Patent No. 12,018,005. The full disclosures of each of WO 2017 / 013498; WO 2019 / 200345; WO 2021 / 158586; WO 2023 / 208133; and U. S. Patent No. 12,018,005 are hereby incorporated by reference herein in their entirety.

[0087] Yet, in another aspect, the Sigma-1 agonist is in a crystal form selected from A2-73 freebase Form I, A2-73 Crystal Form I, A2-73 Crystal Form II, A2-73 Crystal Form III, (-) A2-73 enantiomer, (+) A2-73 enantiomer, A2-73 Freebase Fumarate Salt Form I, A2-73 Freebase Fumarate Salt Form II, A2-73 Freebase Fumarate Salt Form III, A2-73 Freebase Sulfate Form I, A2-73 Freebase Sulfate Form II, A2-73 Freebase Dihydrogen Phosphate Form I, A2-73 Freebase Dihydrogen Phosphate Form II, A2-73 Freebase Mesylate Form I, A2-73 Freebase Mesylate Form II, A2-73 Freebase Oxalate Form I, A2-73 Freebase Oxalate Form II, A2-73 Freebase Oxalate Form III, A2-73 Freebase Benzoate Form I, A2-73 Freebase Edisylate Form I, A2-73 Freebase HBrSalt Form A and A2-73 Freebase HBrSalt Form B, A2-73 Freebase Maleate Salt Form S5, and A2-73 Freebase Maleate Salt Form S6. Details on the A2-73 freebase salts may be found inWO 2019 / 200345. In another aspect, the Sigma-1 agonist is a co-crystal form comprising an A2-73 co-crystal with a tartaric acid, with a citric acid, and / or with a malic acid. In another aspect, the co-crystal is Crystal Form CSII of A2-73 and tartaric acid, Crystal Form CSIII of co-crystal of A2-73 and citric acid, Crystal Form CSIV of co-crystal of A2-73 with malic acid and the co-crystals may be prepared by methods as detailed in WO 2023 / 208133. In another aspect, the Sigma-1 agonist is a racemic co-crystal disclosed and prepared in WO 2025 / 022317. The present disclosure encompasses various polymorphic forms and co-crystal forms of A2-73, A2-73 freebase, A2-73 freebase salts. The polymorphic forms may be prepared by solvent extraction or by supercritical fluid extraction (SCE). The crystal forms prepared from solvent extraction can be found in WO 2019 / 200345. The crystal 41108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center forms prepared by SCE extraction can be found in WO 2017 / 013498. The present disclosure expressly encompasses the A2-73 Crystal Form I, A2-73 Crystal Form II and A2-73 Crystal Form III as prepared and characterized in WO 2017 / 013498. A2-73 Crystal Form I is a crystal with XRPD peaks as shown in FIG.1 ofWO 2017 / 013498. A2-73 Crystal Form I is further characterized by scanning electron microscope (SEM) micrographs as shown in FIGs. 2-3 of WO 2017 / 013498. A2-73 Crystal Form II is a crystal with XRPD peaks as shown in FIG. 4 ofWO 2017 / 013498. A2-73 Crystal Form II is also characterized by FTIR as shown in FIG. 5 of WO 2017 / 013498. The SEM micrographs of A2-73 Crystal Form II is shown in FIG. 7 of WO 2017 / 013498. A2-73 Crystal Form III is a crystal with XRPD peaks as shown in FIG.8 of WO 2017 / 013498. A2-73 Crystal Form III is characterized by FTIR as shown in FIG. 9 of WO 2017 / 013498. A2-73 Crystal Form III is characterized by scanning electron microscope (SEM) micrograph as shown in FIG.11 of WO 2017 / 013498. The present disclosure also encompasses the polymorphic forms of A2-73 free base. The polymorphic form I of A2-73 freebase and its preparation can be found in WO 2019 / 200345. As an example, a crystal form of A2-73 freebase is the crystalline form characterized by XRPD pattern shown in FIG. 16 of WO 2019 / 200345. And such crystalline form possesses particle shapes depicted in FIG. 15 of WO 2019 / 200345. The present disclosure encompasses an enantiomer of A2-73 or a mixture of such enantiomer. The enantiomer may be an (+) or (-) enantiomer. The enantiomer refers to an optical pure ora substantial optical pure entity. The substantial optical pure entity refers to the opposite rotation entity may be presented in an amount of no more than 20%, such as at or about 0.1%, at or about 0.5%, at or about 1 %, at or about 2%, at or about 5%, at or about 8%, at or about 10%, at or about 12%, at or about 15%, at or about 18%, or at or about 20%. The present disclosure also expressly encompasses the (+) and (-) enantiomer disclosed and characterized in WO 2017 / 013496. The full disclosures of each ofWO 2017 / 013496; WO 2017 / 013498; WO 2019 / 200345; WO 2023 / 208133; and WO 2025 / 022317 are hereby incorporated by reference herein in their entirety.

[0088] In various aspect, the Sigma-1 receptor agonist therapy comprises administering to the subject an oral solution comprising A2-73 freebase, a crystal form of A2-73 freebase, a salt of A2-73 freebase, a crystal form of A2-73 freebase salt, A2-73 amorphous form, a crystal form of A2-73, an enantiomer or stereoisomer 42108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center of A2-73, a co-crystal of A2-73, an enantiomer or stereoisomer of A2-73 freebase, an enantiomer or stereoisomer of A2-73 freebase salt. In yet another aspect of the present disclosure, a Sigma-1 receptor agonist is administered to the subject for a period up to 14 weeks, such as about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 9 weeks, about 10 weeks, about 11 weeks, about 12 weeks, about 13 weeks, or about½ weeks. In another aspect, a Sigma-1 receptor agonist is administered to the subject daily, such as once daily, twice daily, and / or 3 times daily. In another aspect, a Sigma-1 receptor agonist is administered to the subject according to an intermittent regimen. In one aspect, the intermittent dosing regimen comprises at least two cycles, each cycle comprising (a) a dosing period during which the Sigma-1 receptor agonist and / or its composition is administered to said patient; and thereafter (b) a resting period. In one aspect, the dosing period and the resting period are of the same duration. In another aspect, the dosing period and the resting period are of different durations. In one aspect, the dosing period and the resting period is in the range of a lower limit of about 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, and 14 days to an upper limit of about 28 days, 27 days, 26 days, 25 days, 24 days, 23 days, 22 days, 21 days, 20 days, 19, days, 18 days, 17 days, 16 days, 15 days, and 14 days.

[0089] In yet another aspect of the present disclosure, a Sigma-1 receptor agonist therapy comprises A2-73 and its pharmaceutical composition thereof. In one aspect, a composition comprising A2-73 or A2-73 freebase in a crystal form and is administered in the amount of about 40 mg to about 60 mg once daily for about 6-11 weeks. In one aspect, the composition of A2-73 is an oral liquid. In another aspect, A2-73 is administered in about 50 mg daily for up to 11 weeks. Alternatively, A2-73 is administered daily in an escalating dose starting from about 10 mg to ending at about 50 mg once daily for 6-11 weeks. Alternatively, A2-73 is administered daily in an escalating dose starting from about 5 mg to 10 mg to ending at about 20 to 30 mg once daily for 6-11 weeks.

[0090] In another aspect, the Sigma-1 receptor agonist may comprise ANAVEX® 19-144 (A19-144), having the chemical name of 1-(2,2-diphenyltetrahydrofuran-3-yl)-N-methylmethanamine hydrochloride. Without being43108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center bound by any theory, A19-144 is reported as a metabolite of A2-73. In the present disclosure A2-73 is a compound, which is subjected to enzymatic oxidation, reduction or hydrolysis under physiological conditions in the living body and is converted to A19-144 of the present disclosure. A19-144 may be in the forms of A19-144 freebase, A19-144 amorphous form, and / or a A19-144 crystal. Such crystal may be prepared by methods and procedures disclosed in WO 2017 / 013498.Without being bound by theory, A19-144 is reported as a metabolite of A2-73. In the present disclosure A2-73 is a compound, which is subjected to enzymatic oxidation, reduction or hydrolysis under physiological conditions in the living body and is converted to A19-144 of the present disclosure. A19-144 may be in the forms of A19-144 freebase, A19-144 amorphous form, or A19-144 crystal. Such crystal may be prepared by methods and procedures disclosed in WO 2017 / 013498. When the active pharmaceutical ingredient is A3-71, a composition may comprise from about 1 mg to about 50 g, from about 0.1 to about 5 g, from about 0.5 g to about 3 g, from about 1 mg to about 55 mg, from about 5 mg to about 30 mg, from about 40 mg to about 60 mg, from about 80 mg to about 120 mg, from about 180 mg to about 220 mg, from about 0.1 g to about 5 g, or from about 0.5 g to about 3 g of A3-71. The active pharmaceutical ingredient may be administered daily, twice daily, or three times daily. The duration of administration may be about 3-6 weeks, 6-11 weeks, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months or chronically. For example, A2-73 orA3-71 may be administered in an amount of about 40 mg to about 60 mg once daily for about 6-11 weeks or is administered in about 50 mg daily for up to 11 weeks. A3-71 or A2-73 may also be administered daily in an escalating dose starting from about 10 mg to ending at about 50 mg once daily. A3-71 or A2-73 may also be administered daily in an escalating dose starting from about 5 mg to about 10 mg to ending at about 20 mg to about 30 mg once daily.

[0091] In another aspect, the Sigma-1 receptor agonist comprises ANAVEX®1-41 (A1-41), having a chemical name of tetrahydro-N, N-dimethyl-5,5-diphenyl-3-furanmethanamine hydrochloride. In another aspect, the Sigma-1 receptor agonist comprises AV1066, having a chemical name of 1 -(3- 4(((1 R,3S,5S)-adamantan-1-yl)(pheny)methyl)propyl)-4-methylpiperazine. In yet another aspect, the Sigma-1 receptor agonist comprises ANAVEX3-71 (A3-71, AF-710B), having chemical name of 1 -[(2S)-2,8-dimethyl-1 -thia-3,8- diazaspiro[4.5]decan-3-yl]- 44108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center 3-(1 H-indol-3-yl)propan-1-one. In another aspect, the Sigma-1 receptor agonist comprises 1-[(2R)-2,8-dimethyl-1-thia-3,8-diazaspiro[4.5]decan-3-yl]-3-(1 H-indol-3-yl)propan-1-one (AF710A). In another aspect, the Sigma-1 receptor agonist comprises 1-[2,8-dimethyl-1-thia-3,8-diazaspiro[4.5]decan-3-yl]-3-(1 H-indol-3-yl)propan-1-one (AF710). In another aspect, the Sigma-1 receptor agonist comprises their amorphous form, a crystal form therefore, an enantiomer thereof, or a prodrug thereof, or a combination thereof.

[0092] In any of the methods, the Sigma-1 receptor agonist therapy comprises administering to the subject the Sigma-1 agonist for a period up to 14 weeks, such as about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 9 weeks, about 10 weeks, about 11 weeks, about 12 weeks, about 13 weeks, or about½ weeks. In another aspect, a Sigma-1 receptor agonist is administered to the subject daily, such as once daily, twice daily, and / or 3 times daily. In another aspect, a Sigma-1 receptor agonist is administered to the subject according to an intermittent regimen. In one aspect, the intermittent dosing regimen comprises at least two cycles, each cycle comprising (a) a dosing period during which the Sigma-1 receptor agonist and / or its composition is administered to said patient; and thereafter (b) a resting period. In one aspect, the dosing period and the resting period are of the same duration. In another aspect, the dosing period and the resting period are of different durations. In one aspect, the dosing period and the resting period is in the range of a lower limit of about 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, and 14 days to an upper limit of about 28 days, 27 days, 26 days, 25 days, 24 days, 23 days, 22 days, 21 days, 20 days, 19, days, 18 days, 17 days, 16 days, 15 days, and 14 days.

[0093] In one aspect, the Sigma-1 receptor agonist is A2-73. When the agonist is A2-73, the therapeutically effective amount of A2-73 can range from about 0.5 mg to about 20 mg, about 1 mg to about 60 mg, about 5 mg to about 30 mg, about 10 mg to about 30 mg; about 5 mg to about 20 mg, about 5 mg to about 10 mg, about 30 mg to about 50 mg, or about 3 mg to about 5 mg. In one aspect, the therapeutically effective amount of A2-73 ranges from about 20 mg to about 60 mg when administered orally. In another aspect, the therapeutically effective amount of A2-73 ranges from about 1 mg to about 20 mg when administered intravenously. In45108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center another aspect, the therapeutically effective amount of A2-73 ranges from about 40 mg to about 60 mg / day. The therapeutically effective amount of A2-73 can range from about 0.5 mg / day to about 100 mg / day, from about 1 to about 60 mg / day, from about 20 to about 50 mg / day, from about 20 to about 30 mg / day, or from about 15 to about 25 mg / day. Administering the anti-neurodegenerative effective amount of A2-73 can provide blood levels of about 10 ng / ml, about 12 ng / ml of A2-73.

[0094] A2-73 can be administered to the subject daily or more than once daily. Further, A2-73 can be administered every 2, 3, 4, 5, 6, 7, 14, or every 30 days. A2-73 can be administered over a period ranging from about 1 day to about 1 year, from about 1 day to about 1 week, from about 3 days to about 1 month, from about 2 weeks to about 6 months, or from about 2 months to about 4 months. A2-73 can also be administered over a period of about 1 day, about 7 days, about 30 days, about 60 days, about 120 days, or about 180 days or more. In some aspects, A2-73 is administered over a period of about 57 weeks, about½8 weeks, about 208 weeks, indefinitely, or until resolution of the condition being treated.

[0095] In an aspect, A2-73 or pharmaceutically acceptable salts thereof can be administered orally to the subject daily in an amount of about 10 mg. In another aspect, A2-73 or pharmaceutically acceptable salts thereof can be administered orally to the subject daily in an amount of about 20 mg. In another aspect, A2-73 or pharmaceutically acceptable salts thereof can be administered to the subject orally daily in an amount of about 30 mg. In another aspect, A2-73 or pharmaceutically acceptable salts thereof can be administered to the subject orally daily in an amount of about 40 mg. In another aspect, A2-73 or pharmaceutically acceptable salts thereof can be administered orally to the subject daily in an amount of about 50 mg.

[0096] In an aspect, a hydrochloride salt of A2-73 can be administered orally to the subject daily in an amount of about 10 mg. In another aspect, a hydrochloride salt of A2-73 can be administered orally to the subject daily in an amount of about 20 mg. In another aspect, a hydrochloride salt of A2-73 can be administered orally to the subject daily in an amount of about 30 mg.

[0097] Other methods of administering the Sigma-1 receptor agonists including, for example A2-73 and its various forms, can be found in, e.g.,WO 2014 / 155138; WO 2016 / 064711; WO 2017 / 013496; WO 2017 / 013498;46108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center WO 2017 / 132127; WO 2018 / 231216; WO 2019 / 079794; U. S. Patent No. 9,750,746; U. S. Patent No. 10,413,519; U. S. Patent No. 10,426,754; U. S. Patent No.10,441,563; U. S. Patent No. 10,507,196; U. S. Patent No. 11,071,723; and U. S. Patent No. 11,617,734; the disclosures of each of which are hereby incorporated by reference herein in their entirety.V. Biological Samples

[0098] Any biological samples suitable for transcriptome or gene analysis can be used in the present disclosure. Non-limiting examples of suitable biological samples include fluid samples, biopsy samples, skin samples, and hair samples. Fluid samples may include blood, serum, plasma, saliva, tears, and lymph. In some aspect, the biological sample is a blood sample, a serum sample, a plasma sample or any transcriptomic sample. Methods of collecting a biological sample from a subject are well known in the art.

[0099] In yet another one aspect of the present disclosure, the biological sample comprises a solid, a semi-solid, a semi-fluid, a fluid, a tissue, or other material collected from a subject. A biological sample may be a fluid biological sample. A fluid biological sample may include fluid and some or all of cells, particles, crystals, and other components in the fluid. Examples of biological samples include but are not limited to, blood, serum, plasma, bone marrow, a nasal swab, a nasopharyngeal wash, saliva, urine, gastric fluid, spinal fluid, tears, stool, mucus, sweat, earwax, oil, a glandular secretion, cerebral spinal fluid, tissue, semen, vaginal fluid, interstitial fluids derived from tumorous tissue, ocular fluids, spinal fluid, a throat swab, breath, hair, finger nails, skin, biopsy, placental fluid, amniotic fluid, cord blood, lymphatic fluids, cavity fluids, sputum, pus, microbiota, meconium, breast milk and / or other secretions or excretions. Biological samples may include nasopharyngeal wash, or other fluid obtained by washing a body cavity or surface of a subject, or by washing a swab following application of the swab to a body cavity or surface of a subject. Nasal swabs, throat swabs, stool samples, hair, fingernail, ear wax, breath, and other solid, semi-solid, or gaseous samples may be processed in an extraction buffer, e.g., for a fixed or variable amount of time. Examples of tissue samples of the subject may include, but are not limited to, connective tissue, muscle 47108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center tissue, nervous tissue, epithelial tissue, cartilage, cancerous sample, or bone. The sample may be obtained from a human or animal. The sample may be obtained from a vertebrate, e.g., a bird, fish, or mammal, such as a rat, a mouse, a pig, an ape, another primate (including humans), a farm animal, a sport animal, or a pet. The sample may be obtained from a living or dead subject. The sample may be obtained fresh from a subject or may have undergone some form of pre-processing, storage, or transport. The terms “blood” and “whole blood” refer to blood as it exists within an animal and as directly obtained from a subject in a blood sample. Blood contains red blood cells, white blood cells, proteins such as albumin, globulins, and clotting factors, salts, water, and other constituents. A blood sample is a fluid biological sample. In one aspect, a biological sample refers to a fluid sample containing suspended cells, cellular material, fungi, antigens, biological factors, immunogens, proteins, and other material of biological origin generally having a size substantially larger than the size of the molecules that comprise the fluid. Biological factors refer to compounds made by living organisms or attached to living organisms that have biological or physiological activities. Biological factors include but are not limited to biological markers, antibodies, cytokines, growth factors, and other peptides, proteins, lipids and carbohydrates produced biologically. Terms “plasma” and “blood plasma” refer to the liquid portion of blood (e.g., a blood sample) that remains after the removal of blood cells. Red blood cells and white blood cells may be removed by centrifugation of a blood sample, leaving plasma above the pelleted cells in the bottom of the centrifuge tube. Plasma retains blood clotting factors and is obtained from anti-coagulated blood samples. A sample of plasma is a fluid biological sample. The terms “serum” and “blood serum” refer to the liquid portion of blood that remains after blood is allowed to clot, and the clot is removed. Serum differs from plasma in that serum lacks clotting factors: since clotting requires fibrin, thrombin, and other proteins, which form and remain part of a blood clot, serum lacks these proteins while plasma contains them. A sample of blood serum is a fluid biological sample. The present disclosure comprises a first biological sample and / or a second biological sample. The first and the second biological sample can be the same type of biological sample but taken at different time. The first and the second biological sample may be a different type of biological sample taken from the same subject. The first and the second biological sample may be different type of biological sample48108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center taken at different time from the same subject. The first and the second biological sample may be the same type of biological sample taken at the same time, but from different subjects. The first and the second biological sample each is independently any biological sample recited above. In one aspect, the first and the second biological sample each independently, comprises a blood serum sample, a plasma sample, or a biological sample suitable for transcriptomic analysis. Transcriptome analysis is the study of the transcriptome, of the complete set of RNA transcripts that are produced by the genome, under specific circumstances or in a specific cell, often using high-throughput methods. The transcriptomic analysis is useful in identifying the functions of genes, and / or identification of pathways that respond to or ameliorate environmental stresses. RNA-Seq can also identify disease-associated gene fusions, single nucleotide polymorphisms and even allele-specific expression. The present disclosure expressly incorporates any biological sample that are suitable and / or capable of being used for transcriptomic analysis. The biological sample may comprise a solid, a semi-solid, a semi-fluid, a fluid, a tissue, or other material collected from a subject. A biological sample may be a fluid biological sample.

[0100] Gene expression measurement includes both DNA and RNA measurements. They may be accomplished by a variety of methods including northern blotting, quantitative real-time PCR (qRT-PCR), nucleic acid microarrays, Luminex microspheres, and nuclease protection assay. Methods of determining a substantially different level of RNA expression are known in the art and include distribution analysis. In some aspects, a substantially different level of RNA expression is determined by normalizing RNA expression values as Transcripts Per Kilobase Million (TPM).

[0101] Over the last decade, there is an avalanche of biomedical data generation and a parallel expansion in computational capabilities to analyze and make sense of these data. Starting with genome sequencing (such as omics) and widely employed deep sequencing technologies, these trends have now taken hold in all omics disciplines and increasingly call for multi-omics integration as well as data interpretation by artificial intelligence technologies. Artificial intelligence (Al) aided-data modeling (Al Modeling) has made the interpretation of large omics data49108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center sets feasible and meaningful. Use of omics and Al Modeling in biomarker discovery can aid clinicians by identifying markers of risk for developing a certain disease, monitoring care, determining prognosis, and developing druggable targets.Combined, Al Modeling has the power to improve patient care, and facilitates the development of precision medicine. In the present disclosure, the Al Modeling not only considers omics data, but also a broad range of other subjects’ data. Such data include, but not limited to, subject’s clinical diagnosis, co-morbidity, co-medication, age, weight, height, and gender. The Al Modeling used in the present disclosure also include trial data, such as doses received, dosing time points, endpoint assessment, and behavior impacts, such as sleep or cognition impacts. Thorough data collection and advanced algorithm, the Al Modeling in the present disclosure can generate and analyze over 24 million relationships and concludes with biomarkers with convincing confidence and p values.VI. Pharmaceutical Formulations

[0102] One aspect of the disclosure encompasses a pharmaceutical formulation for diagnosis and delivery of Sigma-1 receptor agonists. A pharmaceutical formulation comprises a therapeutically effective amount of Sigma-1 receptor agonists and any pharmaceutically acceptable salt(s) thereof.

[0103] Pharmaceutically acceptable salts of the Sigma-1 receptor agonists disclosed herein include, without limitation, acetate, aspartate, benzoate, bitartrate, citrate, formate, gluconate, glucuronate, glutamate, fumarate, hydrochloride, hydrobromide, hydroiodide, hypophosphite, isobutyrate, isocitrate, lactate, malate, maleate, meconate, methylbromide, methanesulfonate, monohydrate, mucate, nitrate, oxalate, phenylpropionate, phosphate, phthalate, propionate, pyruvate, salicylate, stearate, succinate, sulfate, tannate, tartrate, terephthalate, valerate, and the like.

[0104] When the Sigma-1 receptor agonist is A2-73, a formulation can comprise from about 1 mg to about 50 g, from about 0.1 to about 5 g, from about 0.5 g to about 3 g, from about 1 mg to about 55 mg, from about 40 mg to about 60 mg, from about 80 mg to about 120 mg, from about 180 mg to about 220 mg, from about50108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center 0.1g to about 5 g, or from about 0.5 g to about 3 g of A2-73. Formulations comprising A2-73 can be found in, e.g., U. S. Patent No. 9,750,746; U. S. Patent No. 10,413,519; U. S. Patent No. 10,426,754; U. S. Patent No. 10,441,563; U. S. Patent No.10,507,196; U. S. Patent No. 11,071,723; and U. S. Patent No. 11,617,734; the disclosures of each of which are hereby incorporated by reference herein in their entirety.

[0105] Sigma-1 receptor agonists can be formulated and administered to a subject by several different means. For instance, a composition can generally be administered parenteraly, intraperitoneally, intravascularly, transdermally, subcutaneously, or intrapulmonarily in dosage unit formulations containing conventional nontoxic pharmaceutically acceptable adjuvants, carriers, excipients, and vehicles as desired. The term parenteral as used herein includes subcutaneous, intravenous, intramuscular, intrathecal, or intrasternal injection, or infusion techniques. Formulation of pharmaceutical compositions is discussed in, for example, Hoover, John E., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa. (1975), and Liberman, H. A. and Lachman, L., Eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, N. Y. (1980) (each of which are hereby incorporated by reference herein).

[0106] A pharmaceutical formulation comprises one or more pharmaceutically acceptable excipients. Non-limiting examples of excipients include chemical enhancers, humectants, pressure sensitive adhesives, antioxidants, solubilizers, thickening agents, plasticizers, adjuvants, carriers, excipients, vehicles, coatings, and any combinations thereof. One or more excipients can be selected for oral, transdermal, parenteral, intraperitoneal, intravascular, subcutaneous, by inhalation spray, rectal, or intrapulmonary administration.

[0107] A Sigma-1 receptor agonist can in general be formulated for improving patient compliance, preventing a subject from removing the drug-delivery device. For instance, Sigma-1 receptor agonists could be formulated for improved patient compliance and preventing removal of a drug-delivery device by providing formulations for extended delivery. Extended delivery can range for periods ranging from more than one day, to months. This may be especially relevant for patients with compromised cognitive and / or motor-control abilities. Extended delivery for51108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center periods can range from about 1 day to about 1 year, from about 1 day to about 1 week, from about 3 days to about 1 month, from about 2 weeks to about 6 months, or from about 2 months to about 4 months.

[0108] Extended release formulations could be used for substantially continuous delivery of drug at a preselected rate. For example, for crystalline A2-73, the drug can be delivered at a rate of from about 1 mg to about 100 mg / day, from about 40 to about 60 mg / day, or from about 10 to about 30 mg / day. Appropriate amounts of crystalline A2-73 can be readily determined by the ordinarily skilled artisan based upon, for example, the intended duration of administration of the drug by the extended release formulation, the delivery mechanism, the particular formulation, and the relative potency of the drug among other factors.Binders

[0109] Non-limiting examples of binders suitable for the formulations of various aspects include starches, pregelatinized starches, gelatin, polyvinylpyrrolidone, cellulose, methylcellulose, sodium carboxymethylcellulose, ethylcellulose, polyacrylamides, polyvinyloxoazolidone, polyvinylalcohols, C12-C18 fatty acid alcohols, polyethylene glycol, polyols, saccharides, oligosaccharides, polypeptides, oligopeptides, and combinations thereof. The polypeptide may be any arrangement of amino acids ranging from about 100 to about 300,000 Daltons.

[0110] The binder can be introduced into the mixture to be granulated in a solid form, including but not limited to a crystal, a particle, a powder, or any other finely divided solid form known in the art. Alternatively, the binder can be dissolved or suspended in a solvent and sprayed onto the mixture in a granulation device as a binder fluid during granulation.Diluents

[0111] Non-limiting examples of diluents (also referred to as “fillers” or “thinners”) include carbohydrates, inorganic compounds, and biocompatible polymers, such as polyvinylpyrrolidone (PVP). Other non-limiting examples of diluents include dibasic calcium sulfate, tribasic calcium sulfate, starch, calcium carbonate, magnesium carbonate, microcrystalline cellulose, dibasic calcium phosphate, tribasic calcium phosphate, magnesium carbonate, magnesium oxide,52108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center calcium silicate, talc, modified starches, saccharides such as sucrose, dextrose, lactose, microcrystalline cellulose, fructose, xylitol, and sorbitol, polyhydric alcohols; starches; pre-manufactured direct compression diluents; and mixtures of any of the foregoing.Disinteqrants

[0112] Disintegrants can be effervescent or non-effervescent. Non-limiting examples of non-effervescent disintegrants include starches such as corn starch, potato starch, pregelatinized and modified starches thereof, sweeteners, clays, such as bentonite, micro-crystalline cellulose, alginates, sodium starch glycolate, gums such as agar, guar, locust bean, karaya, pecitin, and tragacanth. Suitable effervescent disintegrants include but are not limited to sodium bicarbonate in combination with citric acid, and sodium bicarbonate in combination with tartaric acid.Preservatives

[0113] Non-limiting examples of preservatives include, but are not limited to, ascorbic acid and its salts, ascorbyl palmitate, ascorbyl stearate, anoxomer, N-acetylcysteine, benzyl isothiocyanate, m-aminobenzoic acid, o-aminobenzoic acid, p-aminobenzoic acid (PABA), butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), caffeic acid, canthaxantin, alpha-carotene, beta-carotene, beta-caraotene, beta-apo-carotenoic acid, carnosol, carvacrol, catechins, cetyl gallate, chlorogenic acid, citric acid and its salts, clove extract, coffee bean extract, p-coumaric acid, 3,4-dihydroxybenzoic acid, N, N'-diphenyl-p-phenylenediamine (DPPD), dilauryl thiodipropionate, distearyl thiodipropionate, 2,6-di-tert-butylphenol, dodecyl gallate, edetic acid, ellagic acid, erythorbic acid, sodium erythorbate, esculetin, esculin, 6-ethoxy-1,2-dihydro-2,2,4-trimethylquinoline, ethyl gallate, ethyl maltol, ethylenediaminetetraacetic acid (EDTA), eucalyptus extract, eugenol, ferulic acid, flavonoids (e.g., catechin, epicatechin, epicatechin gallate, epigallocatechin (EGC), epigallocatechin gallate (EGCG), polyphenol epigallocatechin-3-gallate), flavones (e.g., apigenin, chrysin, luteolin), flavonols (e.g., datiscetin, myricetin, daemfero), flavanones, fraxetin, fumaric acid, gallic acid, gentian extract, gluconic acid, glycine, gum guaiacum, hesperetin, alpha-hydroxybenzyl phosphinic acid, hydroxycinammic acid, hydroxyglutaric acid, hydroquinone, N-hydroxysuccinic acid,53108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center hydroxytryrosol, hydroxyurea, rice bran extract, lactic acid and its salts, lecithin, lecithin citrate; R-alpha-lipoic acid, lutein, lycopene, malic acid, maltol, 5-methoxy tryptamine, methyl gallate, monoglyceride citrate; monoisopropyl citrate; morin, betanaphthoflavone, nordihydroguaiaretic acid (NDGA), octyl gallate, oxalic acid, palmityl citrate, phenothiazine, phosphatidylcholine, phosphoric acid, phosphates, phytic acid, phytylubichromel, pimento extract, propyl gallate, polyphosphates, quercetin, trans-resveratrol, rosemary extract, rosmarinic acid, sage extract, sesamol, silymarin, sinapic acid, succinic acid, stearyl citrate, syringic acid, tartaric acid, thymol, tocopherols ( / .e., alpha-, beta-, gamma- and delta-tocopherol), tocotrienols ( / .e., alpha-, beta-, gamma- and delta-tocotrienols), tyrosol, vanilicacid, 2,6-di-tert-butyl-4-hydroxymethylphenol ( / .e., lonox 100), 2,4-(tris-3',5'-bi-tert-butyl-4'-hydroxybenzyl)-mesitylene ( / .e., lonox 330), 2,4,5-trihydroxybutyrophenone, ubiquinone, tertiary butyl hydroquinone (TBHQ), thiodipropionic acid, trihydroxy butyrophenone, tryptamine, tyramine, uric acid, vitamin Kand derivates, vitamin Q10, wheatgerm oil, zeaxanthin, or combinations thereof.Flavor-Modifying Agents

[0114] Suitable flavor-modifying agents include flavorants, taste-masking agents, sweeteners, and the like. Flavorants include, but are not limited to, synthetic flavor oils and flavoring aromatics and / or natural oils, extracts from plants, leaves, flowers, fruits, and combinations thereof. Other non-limiting examples of flavors include cinnamon oils, oil of Wintergreen, peppermint oils, clover oil, hay oil, anise oil, eucalyptus, vanilla, citrus oils such as lemon oil, orange oil, grape and grapefruit oil, fruit essences including apple, peach, pear, strawberry, raspberry, cherry, plum, pineapple, and apricot.

[0115] Taste-masking agents include but are not limited to cellulose hydroxypropyl ethers (HPC) such as Klucel®, Nisswo HPC and PrimaFlo HP22; low-substituted hydroxypropyl ethers (L-HPC); cellulose hydroxypropyl methyl ethers (HPMC) such as Seppifilm-LC, Pharmacoat®, Metolose SR, Opadry YS, PrimaFlo, MP3295A, Benecel MP824, and Benecel MP843; methylcellulose polymers such as Methocel® and Metolose®; Ethylcelluloses (EC) and mixtures thereof such as E461, Ethocel®, Aqualon®-EC, Surelease; Polyvinyl alcohol (PVA) such as Opadry AMB; hydroxyethylcelluloses such as Natrosol®; carboxymethylcelluloses and salts of54108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center carboxymethylcelluloses (CMC) such as Aualon®-CMC; polyvinyl alcohol and polyethylene glycol co-polymers such as Kollicoat IR®; monoglycerides (Myverol), triglycerides (KLX), polyethylene glycols, modified food starch, acrylic polymers and mixtures of acrylic polymers with cellulose ethers such as Eudragit® EPO, Eudragit® RD100, and Eudragit® E100; cellulose acetate phthalate; sepifilms such as mixtures of HPMC and stearic acid, cyclodextrins, and mixtures of these materials. In other aspects, additional taste-masking agents contemplated are those described in U. S. Pat. Nos. 4,851,226; 5,075,114; and 5,876,759, each of which is hereby incorporated by reference in its entirety.

[0116] Non-limiting examples of sweeteners include glucose (corn syrup), dextrose, invert sugar, fructose, and mixtures thereof (when not used as a carrier); saccharin and its various salts such as the sodium salt; dipeptide sweeteners such as aspartame; dihydrochalcone compounds, glycyrrhizin; Stevia rebaudiana (Stevioside); chloro derivatives of sucrose such as sucralose; sugar alcohols such as sorbitol, mannitol, sylitol, hydrogenated starch hydrolysates and the synthetic sweetener 3,6-dihydro-6-methyl-1,2,3-oxathiazin-4-one-2,2-dioxide, particularly the potassium salt (acesulfame-K), and sodium and calcium salts thereof.Lubricants and Glidants

[0117] The lubricant compositions may be utilized to lubricate ingredients that form a pharmaceutical composition. As a glidant, the lubricant facilitates removal of solid dosage forms during the manufacturing process. Non-limiting examples of lubricants and glidants include magnesium stearate, calcium stearate, zinc stearate, hydrogenated vegetable oils, sterotex, polyoxyethylene monostearate, talc, polyethylene glycol, sodium benzoate, sodium lauryl sulfate, magnesium lauryl sulfate, and light mineral oil. The pharmaceutical composition will generally comprise from about 0.01 % to about 10% by weight of a lubricant. In some aspects, the pharmaceutical composition will comprise from about 0.1% to about 5% by weight of a lubricant. In a further aspect, the pharmaceutical composition will comprise from about 0.5% to about 2% by weight of a lubricant.55108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center Dispersants

[0118] Dispersants may include but are not limited to starch, alginic acid, polyvinylpyrrolidones, guar gum, kaolin, bentonite, purified wood cellulose, sodium starch glycolate, isoamorphous silicate, and microcrystalline cellulose as high hydrophilic-lipophilic balance (HLB) emulsifier surfactants.Colorants

[0119] Depending upon the aspect of the disclosure, it may be desirable to include a coloring agent. Suitable color additives include but are not limited to food, drug and cosmetic colors (FD& C), drug and cosmetic colors (D& C), or external drug and cosmetic colors (Ext. D& C). These colors or dyes, along with their corresponding lakes, and certain natural and derived colorants, may be suitable for use in various aspects of the disclosure.pH Modifiers

[0120] Non-limiting examples of pH modifiers include citric acid, acetic acid, tartaric acid, malic acid, fumaric acid, lactic acid, phosphoric acid, sorbic acid, benzoic acid, sodium carbonate and sodium bicarbonate.Chelating Agents

[0121] A chelating agent may be included as an excipient to immobilize oxidative groups, including but not limited to metal ions, in order to inhibit the oxidative degradation of the morphinan by these oxidative groups. Non-limiting examples of chelating agents include lysine, methionine, glycine, gluconate, polysaccharides, glutamate, aspartate, and disodium ethylenediaminetetraacetate (Na2EDTA).Antimicrobial Agents

[0122] An antimicrobial agent may be included as an excipient to minimize the degradation of the compound according to this disclosure by microbial agents, including but not limited to bacteria and fungi. Non-limiting examples of antimicrobials include parabens, chlorobutanol, phenol, calcium propionate, sodium nitrate, sodium nitrite, Na2EDTA, and sulfites including but not limited to sulfur dioxide, sodium bisulfite, and potassium hydrogen sulfite.56108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center Release-Controlling Polymers

[0123] Release-controlling polymers may be included in the various aspects of the solid dosage pharmaceutical compositions incorporating compounds according to this disclosure. In one aspect, the release-controlling polymers may be used as a tablet coating. In other aspects, including but not limited to bilayer tablets, a releasecontrolling polymer may be mixed with the granules and other excipients prior to the formation of a tablet by a known process including but not limited to compression in a tablet mold. Suitable release-controlling polymers include but are not limited to hydrophilic polymers and hydrophobic polymers.

[0124] Suitable hydrophilic release-controlling polymers include, but are not limited to, cellulose acetate, cellulose diacetate, cellulose triacetate, cellulose ethers, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose, microcrystalline cellulose, nitrocellulose, crosslinked starch, agar, casein, chitin, collagen, gelatin, maltose, mannitol, maltodextrin, pectin, pullulan, sorbitol, xylitol, polysaccharides, ammonia alginate, sodium alginate, calcium alginate, potassium alginate, propylene glycol alginate, alginate sodium carmellose, calcium carmellose, carrageenan, fucoidan, furcellaran, arabic gum, carrageens gum, ghafti gum, guar gum, karaya gum, locust bean gum, okra gum, tragacanth gum, scleroglucan gum, xanthan gum, hypnea, laminaran, acrylic polymers, acrylate polymers, carboxyvinyl polymers, copolymers of maleic anhydride and styrene, copolymers of maleic anhydride and ethylene, copolymers of maleic anhydride propylene or copolymers of maleic anhydride isobutylene), crosslinked polyvinyl alcohol and poly N-vinyl-2-pyrrolidone, diesters of polyglucan, polyacrylamides, polyacrylic acid, polyamides, polyethylene glycols, polyethylene oxides, poly(hydroxyalkyl methacrylate), polyvinyl acetate, polyvinyl alcohol, polyvinyl chloride, polystyrenes, polyvinylpyrrolidone, anionic and cationic hydrogels, and combinations thereof.Coatings

[0125] A solid dosage comprising a compound according to this disclosure may comprise a coating, wherein such a coating may control release of the compound, act as a moisture barrier, or buffer or modify pH. A “control releasing coat” or “controlled release coat” as used herein is defined to mean a functional coat which can for example comprise at least one pH independent polymer, pH57108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center dependent polymer (for example enteric or reverse enteric type polymers), soluble polymer, insoluble polymer, lipids, lipidic materials, or combinations thereof. The coating, when applied onto a dosage form, may slow (for example when applied to a normal release matrix dosage form), further slow (for example when applied to a controlled release matrix dosage form) or modify the rate of release of a compound according to this disclosure when applied to an uncoated dosage form. For example, the control releasing coat can be designed such that when the control releasing coat is applied to a dosage form, the dosage form in conjunction with the control releasing coat can exhibit the release of the compound according to this disclosure, such as a “modified-release”, “controlled-release”, “sustained-release”, “extended-release”, “delayed-release”, “prolonged-release,” or combinations thereof. The “control releasing coat” may optionally comprise additional materials that may alter the functionality of the control releasing coat.

[0126] The term “moisture barrier” as used herein is one which impedes or retards the absorption of moisture. Compounds according to this disclosure may be hygroscopic and, as such, may be susceptible to decomposition overtime under highly humid conditions. The proportion of the components of the moisture barrier and the amount of the moisture barrier optionally applied onto the control-releasing coating or onto the core are typically such that the moisture barrier does not fall within the USP definition and requirement for an enteric coat. Suitably, the moisture barrier may comprise an enteric and / or acrylic polymer, suitably an acrylic polymer, optionally a plasticizer, and a permeation enhancer. The permeation enhancer is a hydrophilic substance, which allows water to enter without physical disruption of the coating. The moisture barrier may additionally comprise other conventional inert excipients, which may improve processing of an extended-release formulation.

[0127] Coating and matrix materials which may be used in accordance with the invention are those known in the art for use in controlled-release formulations, such as synthetic polymers of the polyvinyl type, e.g., polyvinylchloride, polyvinylacetate and copolymers thereof, polyvinylalcohol, and polyvinylpyrrolidone; synthetic polymers of the polyethylene type, e.g., polyethylene and polystyrene; acrylic acid polymers; biopolymers or modified biopolymers, such as cellulosic polymers, shellac and gelatin; fats, oils, higher fatty acids and higher alcohols ( / .e.,58108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center acids and alcohols containing alkyl chains of at least 10 carbon atoms), for example aluminum monostearate, cetylalcohol, hydrogenated beef tallow, hydrogenated castor oil, 12-hydroxystearl alcohol, glyceryl mono- or dipalmitate; glyceryl mono-, di-or tristearate; myristyl alcohol, stearic acid, stearyl alcohol, and polyethyleneglycols; waxes; sugars and sugar alcohols.

[0128] The pH-buffering properties of a coating may be strengthened by introducing into the coating substances chosen from a group of compounds usually used in antacid formulations, for example magnesium oxide, hydroxide or carbonate, aluminum or calcium hydroxide, carbonate or silicate; composite aluminum / magnesium compounds, for example Al2O3-6MgO CO2-12H2O, (Mg6Al2(OH)ieCO3-4H2O), MgO ALOs^SiC.nFbO, aluminum bicarbonate coprecipitate or similar compounds; or other pharmaceutically acceptable pH-buffering compounds, for example the sodium, potassium, calcium, magnesium and aluminum salts of phosphoric, carbonic, citric or other suitable, weak, inorganic or organic acids; or suitable organic bases, including basic amino acids; and salts or combinations thereof.

[0129] A pH-dependent coating serves to release the drug in desired areas of the gastrointestinal (Gl) tract, e.g., the stomach or small intestine. When a pH-independent coating is desired, the coating is designed to achieve optimal release regardless of pH-changes in the environmental fluid, e.g., the Gl tract. When the coating is formulated to release a compound according to this disclosure in the intestines (especially the upper small intestines), the coating is often called an “enteric coating”. A pH-dependent coating may include, but is not limited to, acrylic acid polymers and copolymers, for example polymers formed from acrylic acid, methacrylic acid, methyl acrylate, ammonio methylacrylate, ethyl acrylate, methyl methacrylate and / or ethyl methacrylate (e.g., Eudragit™); cellulosic polymers such as hydroxypropyl cellulose, hydroxyethyl cellulose, hydroxypropyl methyl cellulose, methyl cellulose, ethyl cellulose, cellulose acetate, cellulose acetate phthalate (CAP), cellulose acetate trimellitate, hydroxypropylmethyl cellulose phthalate, hydroxypropylmethyl cellulose succinate and carboxymethylcellulose sodium; shellac (purified lac); vinyl polymers and copolymers such as polyvinyl pyrrolidone, polyvinyl59108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center acetate, polyvinylacetate phthalate (PVAP), vinylacetate crotonic acid copolymer, and ethylene-vinyl acetate copolymers; zein; and salts and combinations thereof.VII. Dosage Forms

[0130] One aspect of the disclosure encompasses dosage forms of Sigma-1 receptor agonists. When the agonist is A2-73, for example, the dosage form can comprise from about 1 mg to about 50 g, from about 1 mg to about 500 mg, from about 1 mg to about 100 mg, from about 1 mg to about 500 mg, from about 50 to about 400 mg, from about 75 to about 150 mg, from about 150 to about 200 mg, from about 40 mg to about 60 mg, from about 80 mg to about 120 mg, or from about 180 mg to about 220 mg of A2-73. For instance, the dosage form can comprise 1, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 120, 140, 160, 180, 200, 220, 240, 260, 280, or 300 or more mg of A2-73. In some aspects, dosage forms can comprise from about 1 mg to about 500 mg, or about 1 mg to about 100 mg of A2-73.

[0131] Dosage forms include those formulated for extended or slow release, and those formulated for immediate release. For example, an immediate release dosage form may include a crystalline form of A2-73 as the free base or as a A2-73 salt as disclosed herein. For example, a fast-dissolve oral dosage form may include for example an A2-73 salt, such as the HCI salt. Alternatively, a dosage form may include a crystalline form of A2-73 as the free base or as a A2-73 salt formulated for inhalation drug delivery, either as a dry powder or aerosol spray.

[0132] Dosage forms also include those formulated for topical administration. For instance, a dosage form can be formulated as one or more of a gel, ointment, emulsion, microemulsion, solution, suspension, paste, gel, foam, spray, lotion, or cream. In one aspect, a topical administration dosage form is a transdermal patch. When the dosage form is formulated as a transdermal patch, the transdermal patch can contain from about 40 mg to about 60 mg, from about 80 mg to about 120 mg, or from about 180 mg to about 220 mg of A2-73 freebase in crystalline form.

[0133] Dosage forms can alternatively be formulated for oral administration. Dosage forms formulated for oral administration can be tablets to swallow, chew, or dissolve in water or under the tongue, capsules and chewable capsules, powders,60108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center granules, teas, drops, or liquid medications or syrups. In some aspects, the dosage form is an enteric coated oral formulation.

[0134] When the dosage form is an enteric coated oral formulation, the formulation can comprise from about 0.1 mg to about 60 mg A2-73 freebase, preferably from about 1 mg to about 50 mg A2-73 freebase.

[0135] An enteric coated oral formulation can also contain A2-73 salt in crystalline form. The A2-73 salt can be a fumarate salt, a sulfate salt, a mesylate salt, a dihydrogen phosphate salt, an edisylate salt, a benzoate salt, a hydrochloride salt, and an oxalate salt. In one aspect, the A2-73 salt is a fumarate salt. When the A2-73 salt is a fumarate salt, the enteric coated oral formulation can comprise from about 0.1 to about 100 mg of A2-73 fumarate salt, preferably from about 1 mg to about 55 mg of A2-73 fumarate salt.

[0136] Dosage forms also encompass those formulated for subcutaneous and / or intramuscular injection. For example, an intramuscular dosage form may comprise A2-73 in the free base form, dissolved in an oil matrix for intramuscular injection, or alternatively prepared as a suspension of the free base for intramuscular injection. A dosage form formulated for subcutaneous or intramuscular injection may comprise A2-73 in a salt or free base form as disclosed herein, prepared as microspheres using methods known in the art. Alternatively, A2-73 in free base or salt form may be coated, for example using Atomic Layer Deposition (ALD) techniques, with a thin layer coating such as a coating of zinc oxide, and used in a formulation for subcutaneous or intramuscular injection. Alternatively, A2-73 free base may be dissolved in a biodegradable polymer matrix, and then implanted subcutaneously (or used in a transdermal patch as detailed further below).VIII. Kits

[0137] A further aspect of the present disclosure provides kits comprising one or more reagents for use with a method of the disclosure. The kits may comprise cell growth media, selection media, nucleic acid sequencing and purification reagents, protein purification reagents, buffers, and the like. In one aspect, the kit comprises a Sigma-1 receptor agonist as provided herein (e.g., A2-73) and61108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center components for collecting a biological sample from the subject (e.g., specimen collection containers or vials, swabs, syringes, and the like). The kits provided herein generally include instructions for carrying out the methods detailed herein. Instructions included in the kits may be affixed to packaging material or may be included as a package insert. While the instructions are typically written or printed materials, they are not limited to such. Any medium capable of storing such instructions and communicating them to an end user is contemplated by this disclosure. Such media include, but are not limited to, electronic storage media (e.g., magnetic discs, tapes, cartridges, chips), optical media (e.g., CD ROM), and the like. As used herein, the term “instructions” may include the address of an internet site that provides the instructions.DEFINITIONS

[0138] Unless defined otherwise, all technical and scientific terms used herein have the meaning commonly understood by a person skilled in the art to which this invention belongs. The following references provide one of skill with a general definition of many of the terms used in this invention: Singleton et al., Dictionary of Microbiology and Molecular Biology (2nd ed. 1994); The Cambridge Dictionary of Science and Technology (Walker ed., 1988); The Glossary of Genetics, 5th Ed., R. Rieger et al. (eds.), Springer Verlag (1991); and Hale & Marham, The Harper Collins Dictionary of Biology (1991). As used herein, the following terms have the meanings ascribed to them unless specified otherwise.

[0139] When introducing elements of the present disclosure or the preferred aspects(s) thereof, the articles “a”, “an”, “the” and “said” are intended to mean that there are one or more of the elements. The terms “comprising”, “including” and “having” are intended to be inclusive and mean that there may be additional elements other than the listed elements.

[0140] As various changes could be made in the above-described cells and methods without departing from the scope of the invention, it is intended that all matter contained in the above description and in the examples given below, shall be interpreted as illustrative and not in a limiting sense.62108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center

[0141] The term “comprising” means “including, but not necessarily limited to”; it specifically indicates open-ended inclusion or membership in a so-described combination, group, series and the like. The terms “comprising” and “including” as used herein are inclusive and / or open-ended and do not exclude additional, unrecited elements or method processes. The term “consisting essentially of” is more limiting than “comprising” but not as restrictive as “consisting of.” Specifically, the term “consisting essentially of” limits membership to the specified materials or steps and those that do not materially affect the essential characteristics of the claimed invention.

[0142] As used herein, “expression” includes but is not limited to one or more of the following: transcription of the gene into precursor mRNA; splicing and other processing of the precursor mRNA to produce mature mRNA; mRNA stability; translation of the mature mRNA into protein (including codon usage and tRNA availability); and glycosylation and / or other modifications of the translation product, if required for proper expression and function.

[0143] As used herein, the term “gene” means a segment of DNA that contains all the information for the regulated biosynthesis of an RNA product, including promoters, exons, introns, and other untranslated regions that control expression.

[0144] As used herein, the term “genotype” means an unphased 5' to 3' sequence of nucleotide pairs found at one or more polymorphic sites in a locus on a pair of homologous chromosomes in an individual. As used herein, genotype includes a full-genotype and / or a sub-genotype.

[0145] As used herein, the terms “locus” or “genetic locus” are used interchangeably and refer to a location on a chromosome or DNA molecule corresponding to a gene ora physical or phenotypic feature.

[0146] As used herein, the term “mutant” means any heritable variation from the wild-type that is the result of a mutation, e.g., single nucleotide polymorphisms (“SNP”) and insertions / deletions. The term “mutant” is used interchangeably with the terms “marker”, “biomarker”, and “target” throughout the specification.63108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center

[0147] As used herein, the term “medical condition” includes, but is not limited to, any condition or disease manifested as one or more physical and / or psychological symptoms for which treatment is desirable, and includes previously and newly identified diseases and other disorders.

[0148] As used herein, the term “nucleotide pair” means the nucleotides found at a polymorphic site on the two copies of a chromosome from an individual.

[0149] As used herein, the terms “polymorphism”, “polymorphic variant”, and “polymorphic site” are used interchangeably and refer to a position within a locus at which at least two alternative sequences are found. A polymorphic variant of a gene can lead to the abnormal expression or to the production of an abnormal form of the protein; this abnormality may cause or be associated with disease.

[0150] As used herein, the term “polynucleotide” means any RNA or DNA, which may be unmodified or modified RNA or DNA. Polynucleotides include, without limitation, single- and double-stranded DNA, DNA that is a mixture of single- and double-stranded regions, single- and double-stranded RNA, RNA that is mixture of single- and double-stranded regions, and hybrid molecules comprising DNA and RNA that may be single-stranded or, more typically, double-stranded or a mixture of single- and double-stranded regions. In addition, polynucleotide refers to triplestranded regions comprising RNA or DNA or both RNA and DNA. The term polynucleotide also includes DNAs or RNAs containing one or more modified bases and DNAs or RNAs with backbones modified for stability or for other reasons.

[0151] As used herein, the term “polypeptide” means any polypeptide comprising two or more amino acids joined to each other by peptide bonds or modified peptide bonds, / .e., peptide isosteres. Polypeptide refers to both short chains, commonly referred to as peptides, glycopeptides or oligomers, and to longer chains, generally referred to as proteins. Polypeptides may contain amino acids other than the 20 gene-encoded amino acids. Polypeptides include amino acid sequences modified either by natural processes, such as post-translational processing, or by chemical modification techniques that are well-known in the art. Such modifications are well described in basic texts and in more detailed monographs, as well as in a voluminous research literature.64108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center

[0152] As used herein, the term “single nucleotide polymorphism (SNP)” means nucleotide variability at a single position in the genome, in which two alternative bases occur at appreciable frequency ( / .e., >1%) in the human population. A SNP may occur within a gene or within intergenic regions of the genome. SNP probes according to the invention are oligonucleotides that are complementary to a SNP and its flanking nucleic acid sequence(s).

[0153] As used herein, the term “subject’’ means that preferably the subject is a mammal, such as a human, but can also be an animal, e.g., domestic animals (e.g., dogs, cats and the like), farm animals (e.g., cows, sheep, pigs, horses and the like) and laboratory animals (e.g., cynomolgus monkey, rats, mice, guinea pigs and the like).

[0154] As used herein, the administration of an agent or drug to a subject or patient includes self-administration and the administration by another. It is also to be appreciated that the various modes of treatment or prevention of medical conditions as described are intended to mean “substantial”, which includes total but also less than total treatment or prevention, and wherein some biologically or medically relevant result is achieved.EXAMPLES

[0155] The publications discussed above are provided solely for their disclosure before the filing date of the present application. Nothing herein is to be construed as an admission that the invention is not entitled to antedate such disclosure by virtue of prior invention.

[0156] The following examples are included to demonstrate the disclosure. It should be appreciated by those of skill in the art that the techniques disclosed in the following examples represent techniques discovered by the inventors to function well in the practice of the disclosure. Those of skill in the art should, however, in light of the present disclosure, appreciate that many changes could be made in the disclosure and still obtain a like or similar result without departing from the spirit and scope of the disclosure, therefore all matter set forth is to be interpreted as illustrative and not in a limiting sense.65108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent CenterExample 1. Study design, safety, and tolerability of ANAVEX2-73, and patient characteristics.

[0157] ANAVEX2-73, an agonist of SIGMAR1, has been shown by PET scan to directly bind its target and to modulate cholinergic muscarinic receptors in mice. ANAVEX2-73’s clinical potential in AD was initially assessed in mice, followed by a Phase 1 study in healthy volunteers and a 57-week Phase 2a AD study. This study, a 57-week study (FIG. 1 A), enrolled 32 patients with mild to moderate AD and met its primary endpoint of safety and tolerability (Table 1), leading to a 208-week extension study, termed the 208-week study (FIG. 1A). Patients enrolled in the 2 studies were characterized at baseline and at each visit, including collection of data on clinical descriptors, physical examination, pharmacokinetics and MMSE and ADCS-ADL as efficacy measures (FIG. 1B and Table 3).Table 1. Summary of subject disposition in 57-week study and the 208- week extension study*: number of patients in on-going trial ANAVEX2-73-003 at interim cumulative week 14866108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center Table 3. Baseline CharacteristicsExample 2. Identification of genomic biomarkers.

[0158] Genomic characterization of patients’ tumors is routinely performed in oncology studies to identify and rank genomic biomarkers of disease and therapeutic response. Genetic polymorphisms are associated with variations in treatment response, modulating therapeutic response in a patient population. In this study, Whole Exome Sequencing (WES) and gene expression analysis (RNAseq) were performed at 109-135 weeks (FIG. 1B) for the 21 patients remaining in the extension trial. Genomic data integrated with longitudinal clinical and other patient characteristics including efficacy outcome measures and derived measures of progression, leads to phenotypic and genotypic precision medicine analysis (FIG. 1B, FIG. 1C)67108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center

[0159] A small cohort, with the number of patient observations orders of magnitude smaller than number of attributes, constitutes an analytical challenge. In this study, an unsupervised Formal Concept Analysis method (FCA), based on Galois lattices, as implemented in the KEM software (v.3.6.2), was used to identify and rank phenotypic and genotypic biomarkers. No link is assumed in this analysis between biomarkers and outcomes, enabling a hypothesis free, data-driven, generation of all relations.

[0160] A total of 3,145,630 relations between all available features were explored. Stringent filtering excluded rules containing variables coding for the absence of DNA variants and medium RNA expression groups. Numerical filtering of rules involved the following thresholds: support (n>3), confidence (>50%), lift (>1.2) and p-value (Fisher Exact Test or Mann-Whitney-Wilcoxon < 0.05). The filtering steps led to a subset of 1,019 association rules linking clinical, genomic and transcriptomics patient characteristics with response at multiple time points. A second filtering step focused on relationships with response at week 57 only, raising support > 5, and lift > 1.5, and excluding RNA expression, CYP variants and dose / concentration from Part A. This further reduced the number of rules to 15 associations linked to outcome as measured by Delta (with binarized discretization) or Slope of MMSE or ADCS-ADL at week 57. The 15 rules are presented in order of highest confidence and highest lift. The stringent filtering focused on relationships of response at week 57 (end of Part B), reduced the number of associations to only 15 linked to outcomes as measured by Delta or Slope MMSE or ADCS-ADL (Table 4).Within the 15 relations identified, average blood concentration of ANAVEX2-73 above 4ng / mL in Part B increased the probability of improved MMSE outcome 1.88-fold at week 57 (Table 4). Higher concentrations improved Delta ADCS-ADL scores (p = 0.03) (FIG. 2C). Lower MMSE baseline scores (<20) increased the relative probability (lift) of worse MMSE outcome by 1.5-fold, while higher MMSE baseline scores (>20) increased the relative probability of improved MMSE by 1.62-fold, both at week 57.68108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent CenterTable 4.

[0161] Further focusing on relations displaying a 100% confidence and true for both Delta MMSE and Delta ADCS-ADL, two DNA variants were identified, SIGMAR 1_Q2P and COMT_L146fs. Table 5 shows a summary of integrated data sources. A total of 2,527 features, from 1,152 descriptors, were used for each subject including 837 genomic sequences with amino acid changes, from a total of 27,155 annotated genes; and 185 RNA expression profiles. Patient descriptors are shown in grey and outcomes in pink. c. Description of number of patient descriptors69108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center incorporated in the two analytical steps applied in the study 1) Unsupervised FCA rule-based analysis of response at week 57, 2) Longitudinal confirmation using Mixed Effect modelling of response over 148 weeks incorporating markers found at week 57 to model two groups of ANAVEX2-73 concentrations.Table 5.

[0162] The 3D structural impact of SIGMAR1 Q2P was also investigated. The 3D crystal structure of SIGMAR1 was visualized using VistaPDB 1.0 software (FIG. 3). Each monomer displays an N terminal trans-membrane alpha helix. The N terminus of the protein is believed to face the Cytosol and is believed to interact with other ligands. The identified SIGMAR1 variant (Glutamine (Gin) at position 2 changed into Proline - SIGMAR1_p. Q2P) is located at the N terminus of the protein and may hence have structural and functional implications. The Gin (Q) to Proline (P) alteration impacts the length of the side chain (extended in Gin) and may impact the ability of the amino acid to form an alpha helix. Proline is considered as a “helix breaker” which, in some instances, has been attributed to unwinding the N terminus of a helix. Retrospective exclusion of patients with SIGMAR1_Q2P or COMT_L146fs variants and low baseline MMSE score, improved MMSE and ADCS-ADL scores at week 57. Daily treatment with ANAVEX2-73 for 57 weeks for the entire Phase 2a cohort had effect sizes of 0.57 and 0.18 for Delta MMSE and Delta ADCS-ADL, respectively, when compared to standard of care (Table 6). If patients with the SIGMAR1_Q2P and COMT_L146fs variants and low Baseline MMSE score (9 out of 21 patients) were excluded, effect sizes increased to 1.05 and 0.93 for70108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center MMSE and ADCS-ADL, respectively (Table 6), both considered large’ according to Cohen’s d guidelines.Table 6. Summary of mean Delta MMSE and mean Delta ADCS-ADL scores at week 57, Cohen’s d effect size calculation when compared to standard of care depending on genomic variant status of SIGMAR1_Q2P and COMT_L146fs, and / or baseline MMSE score.Example 3. Validation of results.

[0163] As an internal validation of results at week 57, predictors of response at week 57 were tested with longitudinal data at the interim week 148 in the 208-week study extension study, using regression analyses. This analysis included 21 patients with genomic data and 287 observations for 14 time points (Table 7).ANAVEX2-73 concentration in Part B, identified as predictor of outcome at week 57, was used to define 2 arms for the MMRM-LME model. In addition to parameters 71108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center identified at week 57 (ANAVEX2-73 concentration levels, Baseline MMSE score, SIGMAR 1_Q2P and COMT_L146fs variants), age, sex, APOE £4 status, and donepezil co-medication were included in the MMRM-LME models of Delta MMSE and Delta ADCS-ADL. This analysis showed that the higher ANAVEX2-73 concentration arm had favorable differences of 78% and 88% in adjusted MMSE and adjusted ADCS-ADL, respectively, when compared with the low / medium arm over 148 weeks (p-values < 0.0008 and < 0.0001, respectively). In addition to time, APOE e4 status (p< 0.0001) and ANAVEX2-73 mean concentration were significant predictors (FIGS.4A-4B and Table 8 and Table 6A). Furthermore, SIGMAR1_Q2P, COMT_ L146fs, and APOE4 £4 status interactions with time were significant variables. Unadjusted measures corroborated these results; patients with positive biomarkers of response had better Delta ADCS-ADL over 148 weeks than those without the biomarkers or than reference populations receiving standard of care (FIG. 5 and Table 9). Two patients showed exceptional therapeutic response during the longitudinal study (148 weeks period). Both subjects had SIGMAR1 wild type, high mean concentration of ANAVEX2-73 in plasma and baseline MMSE>20 (FIG. 6). Clinical benefits reported included improvements in mood and alertness and increased independent activity.Table 6AMMRM-LME model output for Delta ADCS-ADL outcomePatient descriptor (F test)Time <0.001AV-2-73 Concentration (= High) 0.38Gender (=Male) 0.535APOE4 status (=True) 0.643Age (=Low) 0.946Donepezil treatment (=T rue) 0.959Time: AV2-73 Concentration (=High) <0.0001Time: APOE4 status (=True) <0.0001AV2-73 Concentration (=High): APOE4 status (=True) 0.762Baseline MMSE (=Low) 0.315SIGMAR1 Q2P variant (=True) 0704COMT variant (=True) 0.523SIGMAR1 variant Q2P (=True): Time <0.01COMT variant (=True): Time <0.01SIGMAR1 Q2P variant (=True): AV2-73 Concentration (=High) 0.71672108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent CenterTable 7MMRM*LME model output for Delta MMSE outcomePatient descriptorP valu®(F test}Time <0.0001Age (=Low) 0.801 Gender =Male) 0.146 Baseline MMSE (=Low) 0.129APOE4 status (-True) 0.812 SIGMAR1 Q2P variant (=True) 0.292AV2-73 Cone (=High) 0.726Time: APOE4 status (“True) <0.0001Time: V2-73 Cone (=High) <0.0001 SIGMAR1 Q2P variant (=True): AV2-73 Cone £=High) 0.684Table 8MMRM-LME model output for Delta ADCS-ADL outcome...... P-value (F Patient descriptor.xtest) Time <0.001AV-2-73 Concentration (= High) 0.38 Gender {=Male) 0.535 APOE4 status (=True) 0.643Age (= Low) 0.948 Donepezii treatment (= rue) 0.959 Time: AV2-73 Concentration (=High) <0.0001 Time: APOE4 status (-True) <0.0001 AV2-73 Concentration ( = High): APOE4 status (“True) 0.762Baseline MMSE {=Low) 0.315 SIGMAR1 Q2P variant (=True) 0.704 COMT variant (=True) 0.523 SIGMAR1 variant Q2P (-True): Time <0.01 COMT variant (=True): Time <0.01 SIGMAR1 Q2P variant (=True): AV2-73 Concentration (=High) 0.71673108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center Table 9. Summary of the mean and standard deviations of different patient groups unadjusted change in ADCS-ADL scores at½ time points (Week 0, 5, 12, 26, 36, 48, 57, 83, 96, 109, 122, 135 and 148). All patients in ANAVEX2-73 study are presented along with each patient subgroup depending on inclusion criteria. The standard of care mean change from baseline for ADCS-ADL scores were obtained from literature with -6.7 point change in one year and -10.5 change in 18 months / 70 weeks.74108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent CenterDiscussion for Examples 1-3

[0164] This study is, to the best of our knowledge, the first reported genomewide search for biomarkers associated with drug response in AD. The approach we used was different from standard classification or searches for “signatures”. KEM framework-based FCA was used as a method for identification and ranking of predictors of response by association rules consistent with the clinical studies’ data. As shown in this work, the FCA machine learning methodology enabled exploring a very large combinatorial space, typically 109and larger; extracting smaller sets, typically 2-3x106; and finally focusing on dozens of important relations. In contrast with “black box” numerical methods, this approach allows the identification of patients verifying each identified rule (as shown in Tables 4 and 5) and puts in place objective and explicit filtering strategies to select the most relevant biomarkers.

[0165] Negative results generated around the “amyloid hypothesis” have encouraged the AD community to hypothesize that the Ap plaques may appear at an advanced stage of the disease, which may be beyond the reach of existing anti-BACE or anti-tau interventions. However, characterizing the disease at earlier stages has also been challenging, since reliable genomic markers that may be prognostic of the onset of MCI caused by AD have not yet been identified. APOE status alone does not have sufficient predictive power, because only about 25% of the population carry the APOE4 genotype (but see Examples below). Studies have identified a genetic variant, TOMM40 that could be used for the early detection of Mild Cognitive Impairment (MCI). This hypothesis was tested in the TOMORROW study. However, the TOMORROW study including 3,494 patients (out of 24,136 screened) was not able to reach its primary objective of biomarker qualification. In this context, the search for alternative druggable targets for AD is becoming a priority75108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center and SIGMAR1 targeted by ANAVEX2-73 may be an important drug target to maintain cellular homeostasis, and which may potentially delay or halt neurodegeneration and even allow compensatory responses.

[0166] This paper presents a data-driven, unbiased analysis of data from two clinical studies, which enabled the identification of parameters associated with improved outcome. By searching for and selecting biomarkers that are consistently associated with clinical response over multiple time points and multiple endpoints, a reliable identification and ranking of patient selection markers is possible. It is noteworthy that the approach described in this study, without any a priori hypotheses, identifies in addition to a COMT variant (rs113895332 / rs61143203), ANAVEX2-73 mean concentration, baseline MMSE and a specific variant of the SIGMAR1 gene (rs1800866) as the top 20 drivers of clinical response, amongst the >3 million retained subset of rules. Low baseline MMSE is thought to be associated with an AD stage that has progressed beyond the reach of a number of therapeutic approaches. SIGMAR1 is the confirmed in vivo target of ANAVEX2-73. COMT is a gene that has been previously described as involved in memory and other neurobehavioral functions.

[0167] The MMRM-LME analysis at interim week 148 showed a significant difference in response between the 2 concentration arms related to adjusted mean Delta ADCS-ADL (88%) and adjusted mean Delta MMSE (78%). It is noted that these differences are significant and quite large. These values compare favorably with recent results obtained in other studies for e.g. data presented at CTAD 2018 for BAN2401 after 72 weeks: 47% in adjusted mean change from baseline in ADAS-Cog and 30% adjusted mean change in ADCOMS54, where about 18% of the changes may be due to the imbalance of the APOE4 status in the respective arms.

[0168] An analysis of 39,974 DNA variants (Table 10) showed SIGMAR1_Q2P and COMT_L146fs were retained in the top associations. Due to limited data availability, the standard of care measure of Delta MMSE and Delta ADCS-ADL obtained from the literature to calculate effect size corresponded to 52 weeks instead of 57 weeks, the actual time point used in this study. Finally, the RNA analysis was limited due to collection time (109-135 weeks) and no baseline transcriptome measure was available for comparison. However, an exploratory76108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center analysis found that high SIGMAR1 expression was associated with better outcome (Table 11).Table 1077108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center78108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center79108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center80108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center81108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center82108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent CenterTable 11. Association rules linking SIGMAR1 RNA expression with MMSE and ADCS-ADL endpoints between weeks 57 and 148. Highlighted in blue are time points when blood was collected for RNA expression analysis.83108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Centernx: number of antecedent (patients in RNA expression group) observations ny: number of consequent (Endpoint) observations Deriv.: Calculation used for derived endpoint descriptor

[0169] The methodology that has been described in this work opens the possibility of using data-driven unbiased biomarker identification early in the drug development process. The “white box” and systematic approach of Formal Concept Analysis may be an ideal match for the analysis of early data, where a machine learning platform may explain each finding and be essential for identifying and ranking patient selection biomarkers for therapeutic response. These data analytic methods could become cornerstones for precision medicine drug discovery and development.84108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center Methods for Examples 1-3(a) Study design

[0170] The present study integrates and analyzes data and samples from two consecutive clinical trials: 57-week study and 208-week study. 57-week study was a multicenter Phase 2a clinical trial of the drug ANAVEX2-73 that recruited a sample of 32 subjects with mild-to-moderate AD. It consisted of two parts: Part A was a simple randomized, open-label, two-period, cross-over, adaptive trial lasting up to 5 weeks. Each period involved one specific administration route and two possible dose levels: in one period the oral dose (30mg or 50mg) was administrated and in the other period an intravenous (IV) form (3mg or 5mg) was administered, making a total of 8 possible dose administration schemes in Part A (details available in FIG. 2A). The two periods were separated by a wash-out period of 12 days where no dose was given.

[0171] Part B, following immediately after Part A, was an open-label extension of the oral daily dose for an additional 52 weeks. The primary endpoint of 57-week study was to establish the safety and tolerability of the drug. Secondary endpoints aimed at establishing the relationship between dosing regimen and pharmacodynamics-exploratory efficacy outcomes of cognition and function.Cognition was assessed by the Mini Mental State Examination (MMSE), while functional abilities by the Alzheimer’s Disease Co-operative Study - Activities of Daily Living Inventory (ADCS-ADL). Additional cognitive measures included the Cogstate Brief Battery (CBB), and functional measures of Electroencephalography and Event-Related Potential (EEG and ERP). Part B continued an adaptive oral administration scheme. Table 12 summarizes mean oral dose administration during the 57-week study study.Table 12. Summary statistics and binning thresholds of ANAVEX2-73 mean dose administration in mg at three different time periods: Part A1 (Day1-2: within 2 days of85108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center first oral dose), Part A2 (Day3-12: after the first two days of first oral dose and before beginning of Part B) and Part B (Week 5 to Week 57). (N=32 patients).Mean Dose (mg)PartAI PartA2 Part B

[0172] Due to favorable safety and tolerability profile of the 57-week study (Table 1), and in response to requests from patients and caregivers, a 208-week open-label extension study (208-week study) was carried out immediately after completion of 57-week study Part B. 208-week study enrolled the 24 subjects who completed Part B and applied similarly an adaptive daily oral administration scheme. ANAVEX2-73-003 involves continuous evaluation of safety and efficacy as measured by the MMSE and ADCS-ADL. At week 148 (interim time point for analyses), data on the subjects remaining in the study (n=21) was analyzed for efficacy outcomes. This report was drafted using data available at week 148, while ANAVEX2-73-003 is still ongoing.(b) Subjects

[0173] A total of 32 subjects with mild to moderate AD, between 50 and 85 years of age, were enrolled across three sites in Australia from December 2014 through September 2015. Their diagnoses were consistent with Diagnostic and86108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center Statistical Manual of Mental Disorders 4th edition (DSM-IV) and National Institute of Neurological and Communicative Disorders and Stroke / Alzheimer’s Disease and Related Disorders Association criteria, and included evaluation of MRI and / or CT brain scans. The demographics and baseline characteristics of the 32 participants are summarized in Table 3 along with the group of participating subjects with genomic data collected after protocol amendment and patients consent (N=21). Table 1 presents patient disposition. Seven patients discontinued during the 57-week 57-week study, thereof six were withdrawal of consent and one due to an Adverse Event (AE). And a total of three patients discontinued during 208-week study up to cumulative 148 interim week of the on-going extension study.

[0174] The following assessment scores and criteria were recorded at baseline and at each visit: MMSE, ADCS-ADL, Hamilton Depression Rating Scale (HAM-D) assessment, Rosen Modified Hachinski Ischemic Score (RmHis), CBB, and EEG / ERP. CBB subtests consisted of six different sub-scores: Detection (DET; Processing Speed), Identification (IDN; Attention / Vigilance), One-Card-Learning (OCL; Visual Recognition Memory), One-Back (OBK; Working Memory), International Shopping List (ISLT; Verbal Learning), and International Shopping List delayed recall (ISRL; Verbal Memory).(c) Study endpointsA. Safety endpoints

[0175] Safety was evaluated by assessment of incidence and severity of adverse events, vital signs, physical examination, clinical laboratory parameters (hematology, serum chemistry and urinalysis), and electrocardiogram (data not shown).B. Efficacy outcomes

[0176] Exploratory clinical efficacy in this study was defined as change from baseline of each patient’s cognitive (MMSE) and functional ability (ADCS-ADL) scores. The MMSE57 is a widely used, validated instrument in clinical AD research, including as a cognitive outcome measure in drug trials. The range of the scale is from 0 to 30, with a higher score indicating higher cognitive function. The ADCS-ADL58 is undertaken in a structured caregiver interview format and assesses87108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center performance in activities of daily living of patients with AD. The scale is from 0 to 78, with a higher score indicating greater function. The ADCS-ADL has shown to have good test-retest reliability59. The MMSE and the ADCS-ADL were assessed during pre-specified clinical site visits across 14 different time points during the 57-week study (Weeks 0 and 5 in Part A; Weeks 17, 31, 41,53, and 57 in Part B), and 208-week study (Weeks 70, 83, 96, 109, 122, 135, and 148) studies.

[0177] Corresponding Delta scores were calculated using the difference between values at each visit and at baseline for each individual patient, with a focus on Week 57. In addition, to lower the impact of noise in MMSE and ADCS-ADL scores, a Slope was estimated using a linear regression across all available time points for each individual patient. The derived scores describe the progression of exploratory efficacy data from baseline at several specific time points (Deltas) and over a set period of time (Slope). Including sub-scores and derived measures from the same test score did not have an impact on the analytical method applied in this study since it treated and described each relationship independently.

[0178] Table 13 summarize Delta scores, with respect to baseline, for the MMSE and the ADCS-ADL at 13 time points in 57-week study (Weeks 5, 12, 26, 36, 48, and 57) and in ANAVEX2-73-003 (Weeks 70, 83, 96, 109, 122, 135, and 148). Summary statistics of Slopes for MMSE and ADCS-ADL are illustrated in Table 13.88108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center Table 13Law group High groupPatient baseline characteristicThreshold Count Threshold CountAge < 70 16 > 70 16 BMi £ 25 13 > 25 19 Diastolic Blood Pressure £ 84 22 > 84 10Systolic B ood Pressure < 549 24 > 14S 8Heart Rate £ 74 28 > 74 4 Respiration Rate S 15 28 > 15 4Pulse Oximetry & 96 3 > 96 26Body Temperature s' 36.3 14 > 36.3 18C. Pharmacokinetics

[0179] A pharmacokinetics analysis was performed during Part A only. Blood samples were drawn at 12 and 11 time points after the first oral or IV administration during the first and second periods of Part A, respectively. Pharmacokinetic parameters included in the analysis consisted of maximum plasma drug concentration (Cmax), time to maximum plasma concentration during a dosing interval (tmax), area under the curve from the time of dosing to the last measurable concentration (AUCo-t), mean elimination half-life (t½), and the total body clearance for extravascular administration divided by the fraction of dose absorbed (CL / F). FIG. 2B shows AUCo-inf after first administration (24 first hours).

[0180] Steady-state plasma levels of ANAVEX2-73 and its metabolite ANAVEX19-144 were assessed in Part B through blood samples collection at prespecified time points (Weeks 12, 26, 36 and 48). Mean concentration values in ng / mL of each patient were then averaged. Table 14 illustrate mean plasma concentrations of ANAVEX2-73 and its metabolite ANAVEX19-144 during the 57-week study study.Table 14. Summary statistics and binning thresholds of plasma concentrations of ANAVEX2-73 and its metabolite ANAVEX 19-144 at three time periods: Part A1 (Day1-2: within 2 days of first oral dose), Part A2 (Day3-12: after the first two days of89108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center first oral dose and before beginning of Part B) and Part B (Week 5 to Week 57). (N=32 patients).

[0181] The relationship between dose and plasma concentration of ANAVEX2-73 was assessed during a Phase 1 study, and can be assumed to be linear in the 1-60 mg range in male healthy subjects. The aforementioned dosedependent increase of plasma concentration of ANAVEX2-73 and metabolite has been also observed in the present study, as illustrated in FIG. 2B.D. DNA and RNA extraction and sequencing

[0182] Blood samples were collected from all still participating 21 subjects, between weeks 103-135, depending on the visit date after protocol amendment and approval and patients consent during the ANAVEX2-73-003 extension study. These samples enabled DNA (Whole Exome) and RNA Next Generation Sequencing (NGS) data analysis. 408,551 DNA variants were identified within the cohort. Of these, 39,974 high or moderate impact variants driving an amino acid modification in the protein were kept; RNA expression values were normalized as Transcripts Per Kilobase Million (TPM).E. Focused gene subset

[0183] 243 genes were selected from the 27,155 mapped sequences obtained from NGS analysis introduced above (shown in FIGS. 10A-10C). 102 genes were selected on the basis of their involvement in neurodegenerative diseases (FIG. 10A).90108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center 20 genes were selected as part of SIGMAR1 ’s functional interactome (FIGS. 10C-10D), based on a confidence score of 0.150 obtained from the STRING database (January 2018). Moreover, 113 genes of the cytochrome P450s gene family and 10 genes from the methyltransferase gene family were also added (FIG. 10B).F. Data analysis

[0184] A data analysis using association rule-based FCA was performed to identify markers of response to treatment at week 57. In a subsequent step, confirmatory analyses of identified markers of response at½8 weeks was performed using linear mixed effects. FIG. 1B illustrates the different types of data used in the analyses, whereas FIG. 1C displays the two different types of analyses for identification and validation of markers of drug response.(d) Association rules and the KEM® platform

[0185] The KEM® (Knowledge Extraction and Management) v 3.6.2 software was used to systematically extract relations between all variables collected. Based on Galois lattices theory, or Formal Concept Analysis (FCA), KEM® identifies groups of objects with shared properties and generates corresponding association rules. Association rules have been developed to detect patterns and relationships in large heterogeneous databases. This methodology was introduced by Agrawal and Sirikant and it has been successfully used in different domains including: market basket analysis, drug discovery, and genomic characterization of complex diseases. The method has also been successfully used in drug efficacy and patient stratification studies. Association rules may reflect the presence of a causal relationship between two variables. However, the presence of a statistically significant association is neither necessary nor sufficient to indicate the presence of such causal relationships. Hence, the use of the quality measures described below.

[0186] An association rule is an implication such as X-> Y, with X being the antecedent (or a combination of antecedents; also termed left side) and Y the consequent (or combination of consequents; also termed right side) of the rule, that allows inferring X as an explanation of Y. Considering a dataset composed of N patients, nx, ny and nxy, respectively are the numbers of patients satisfying the antecedent(s) X of a rule, the consequent(s) Y and both parts of the rule,91108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center respectively. For any given rule, it is possible to identify specifically the nxypatients matching this rule.

[0187] Six main quality measures are used to describe and rank association rules, namely: Support, Confidence, Lift, two P-values measures of significance and Cohen’s d a measure of effect size. / . Support, confidence, and lift

[0188] Support is defined as the number of subjects for which the association has been observed:SupportfX -> ) = nXY

[0189] Confidence is defined as the percentage of the characterized patients verifying the rule:

[0190] The Lift (relative probability) is the ratio of observed support to that expected if X and Y were independent. It measures the performance of a rule to identify a subgroup from a larger population:N X nXYLift ( X T) =nxx ny / / . Significance tests

[0191] P-values from two statistical tests were calculated for each association rule. A right-sided Fisher’s exact test to account for the relationship of the rule, for categorical data and suitable for small sample sizes. And a two-sided Mann-Whitney U test, for comparing continuous outcome measures per descriptor group. Statistical significance was judged for both tests at a level of 0.05.Hi. Effect size

[0192] Cohen’s d index is calculated for each association to quantify the effect size, from which the number of patients to be included to determine a significant relationship in future studies can be calculated. Cohen’s d indicates the standardized difference between two population means.92108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center

[0193] In this study, the outcomes were defined by Delta and Slope values of MMSE and ADCS-ADL. Control group values for calculating effect size were obtained combining trial results available in the AD literature receiving standard of care: the control group mean Delta MMSE at 52 weeks was defined as -3.7 with a standard deviation of 3.5, and mean Delta ADCS-ADL at 52 weeks was -6.7 with a standard deviation of 8.0.

[0194] Continuous variables such as age were discretized using distribution (see summary of thresholds in Table 13). For instance, MMSE at baseline was discretized in two groups, ‘Low’ (defined as baseline MMSE <20) and ‘High’ (defined as baseline MMSE >20). It is noted that the value “20” is not a predefined limit:MMSE baseline score threshold “<20” corresponded to the median of the observed values of all included patients.(e) Linear mixed effect models

[0195] Application of Mixed effect Models for Repeated Measures (MMRM) to model continuous primary endpoints in longitudinal clinical trials and specifically to analyze the outcome of AD clinical studies has become common practice.

[0196] The longitudinal relevance of biomarkers identified at week 57 was assessed at week 148 using MMRM, by allocating subjects to one of two arms (High versus Low / Medium) based on ANAVEX2-73 mean blood concentration in Part B. Given the small cohort size, time was treated as a continuous variable in the analyses to avoid over-parameterization and improve robustness. Therefore, a linear component for mixed-effect models (LME) was applied to model time. MMRM-LME, as implemented in R statistical software, was applied to model Delta MMSE and ADCS-ADL scores with ANAVEX2-73 concentration levels over time as well as other identified descriptors of response.Example 4. DNA & RNA extraction and analysis

[0197] A total of 27,155 annotated genomic locations corresponding to genes were identified in AD patient sequences, including 616 pathways. An initial panel of 243 genes were defined. The panel consisted of 102 genes implicated in neuro-degenerative diseases, 20 genes connected with the SIGMAR1 functional93108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center interactome and 12 methyltransferases genes and 111 CYP genes linked to drug metabolism (Table 15).94108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center95108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center96108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center97108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center

[0198] A distribution analysis was performed using TPM (Transcripts Per kilobase Million) values for each RNA gene expression, defining 3 equal size bins: Low, Medium, High.

[0199] This is calculated as follows:106

[0200] noting Xi. read count of the mRNA considered; and

[0201] / z: length of the mRNA considered.

[0202] For COMT, COMT TPM (COMTt) a level of 10.1 or below is considered Low, 10.1 > COMTt >=15.2 Medium and COMTt > 15.2 High.

[0203] For KANSL1, KANSL1 TPM (KANSLIt) a level of 16.9 or below is considered Low, 16.9> KANSLIt >=27.5 Medium and KANSLIt > 27.5 High.

[0204] For SIGMAR 1, SIGMAR1 TPM (SIGMARIt) a level of 12.7 or below is considered Low, 12.7> SIGMARIt >=19 Medium and SIGMARIt > 19 High.

[0205] Focusing on the initial panel of 243 genes, 185 RNA gene expressions and 837 DNA alterations, at the protein level, were shown to be different for at least one subject out of the total set of 21 subjects. Subjects were further characterized using an additional set of 212 non-genomic characteristics consisting of medical98108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center history, concomitant medication and other baseline characteristics. A total of 40 end points were defined as follows:

[0206] ► Mini-Mental State Examination (“MMSE”) and Alzheimer's Disease Cooperative Study - Activities of Daily Living (“ADCS-ADL”) endpoints were extracted from the data collected in ANAVEX 2-73-002.

[0207] ► Delta MMSE and ADCS-ADL were calculated using the difference between values collected at 6 time points and at baseline.

[0208] ► Similarly, a slope was calculated integrating multiple values. The slope is calculated using a linear regression across all available time-points for each individual patient. The slope calculation has the advantage of smoothing the variability across observed values of these scores.

[0209] ► A distribution analysis was performed on the Deltas calculated for both MMSE and ADCS-ADL (FIGS. 7A-7B). Patients were divided into categories based on third-tiles:

[0210] - Delta ADCS-ADL: High (> -1) and Low (<-2);

[0211] - Delta MMSE: High (> -1) and Low (<-2).

[0212] The Medium and Low bins were combined to define a new Low bin. The High bin corresponds to the highest third-tile values. Slope between baseline and week 57 for both MMSE and ADCS-ADL, respectively, was considered as a binary endpoint. Responders were defined as having a positive or null slope. Non responders were defined as displaying a negative slope.

[0213] The study contained a total of 2,481 features for each patient. KEM® v 3.6.2 (Knowledge Extraction and Management, available from Ariana Pharmaceuticals, SA) was used to analyze the available data. 3,040,276 relations were generated and analyzed. The relations were analyzed using a set metrics to evaluate their strength:

[0214] - Support (n) shall mean the number of subjects for which the Association has been observed.99108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center

[0215] - Lift (relative probability) shall mean the frequency of the Association divided by the frequency of the Condition. This parameter measures the enrichment provided by the Condition.

[0216] - Confidence shall mean the probability of Association when the Condition is True

[0217] - Cohen’s d is an effect size used to indicate the standardized difference between two means. From the association rules we identified some genetic features linked with improved or worsened outcomes (delta MMSE or delta ADCSADL). For each feature we defined Groupl by patients bearing the desirable feature (if it was linked with improved outcome), or by patients without the feature (if it was linked with worsening outcome). MgrOupi and SDgrOupi are respectively the mean and the standard deviation of the outcome for the group 1. MgrOup2 and SDgr0UP2 are the mean and standard deviation referent values of the outcome, known in the art, for a group of patient with mild Alzheimer disease.Table 16. Interpretation of effect sizesuggested by Cohen and Sawilowsky.Effect size d ReferenceVery small 0.01 Sawilowsky, 2009Small 0.20 Cohen, 1988Medium 0.50 Cohen, 1988Large 0.80 Cohen, 1988Very large 1.20 Sawilowsky, 2009Huge 2.0 Sawilowsky, 2009

[0218] - P-value (Fisher’s exact test, greater one-tail) evaluates the statistical significance of difference in a 2x2 contingency table.100108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center

[0219] - Mann- Whitney- Wilcoxon (MWWilc.) is a non-parametric test of the null hypothesis that the distributions in both populations are equal. The MWWilc test ranks values of the union of two populations and compares the sum of their ranks between the two populations. We compare the distribution of the outcome between patients with the feature and patients without the feature.

[0220] - MAF (Minor allele frequency): refers to the frequency at which the mutation occurs in the general population from the 1000 Genomes database.

[0221] The filtering of the relations was performed in 2 steps: a semantic filtering (1) and a numerical filtering (2). At the semantic filtering step, parameters coding for the absence of DNA mutation, medium RNA expression bins, as well as Slope MMSE and Slope ADCS-ADL were excluded. At the numerical filtering step, only relations satisfying metrics based on support (n>3), confidence (>50%), relative probability (lift>= 1.2) and p-value (Fisher or Man Whitney Wilcoxon < 0.05) were retained.

[0222] The filtering steps (1) and (2) leads to a subset of 509 association rules linking Base Line clinical and genomic patient characteristics and RNA expression with response at multiple time points.

[0223] Focusing on response at week 57 only, this set was further reduced to 42 associations linking DNA mutations, 27 RNA expression, and 9 involving other parameters at baseline.

[0224] The 42 association rules linking the presence of a DNA mutation with response correspond to 22 CYP proteins and 20 non CYP proteins from the gene panel defined in Table 15. The 20 non CYP relations correspond to 12 non-redundant SNPs associated with worsened response at week 57 (Table 18) and 2 associated with improved response at week 57 (Table 17).101108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center

[0225] The Table 17 data was extracted from the systematic analysis of genomic data (185 gene panel and 837 mutations). It identifies 2 SNP variants significantly linked with improved outcome as measured by delta MMSE or delta ADCS-ADL at week 57: these are HLA-DRB1_p. LysThr134LysAsp and DPYD_p.l543V (re1801159). Rules: n>3 patients, Conf. > 0.5, Lift >= 1.2, p-Value (Fisher Test or MWWilc.) < 0.05.Table 17. Presence of HLA-DRB1-P. LysThr134LysAsp or DPYD V 543 (DPYD_p.l543V) mutations were associated with improved ADCS-ADL or MMSE Delta (High) at week 57 versus baseline.Table 18. 10 SNP variants significantly linked with worsened outcome as measured by delta MMSE or delta ADCS-ADL at week 57. Rules: n>3 patients; Conf. > 0,5; Lift >= 1,2; p-Value (Fisher Test or MWWilc.) < 0.05102108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center103108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center

[0226] Presence of altered COMT (stop after L146), or DHCR7 T220 (DHCR7_p. M220T), or HLA-DRB1 T244 orT66 orS89 (HLA-DRB1_p. A244T or S66Y orY89S), or KANSL1 Pro304 / 946 / 1010 (KANSL1_p. P1010L / P304L / P946L) or MS4A6E T59 (MS4A6E.p. M59T), or RIN3 R 215 (RIN3_p. H215R) orSIGMARI Pro2 (SIGMAR1_p. Q2P) mutations are associated with ADCS-ADL or MMSE Delta Low at week 57 versus baseline.

[0227] Table 19 presents data on three mutations (SIGMAR1 Q2P;COMT_p. L146fs; KANSL1_p. P1010L / P304L / P946L) associated with both MMSE and ADCS-ADL delta at week 57. Systematic analysis of the genomic data (185 gene panel and 837 mutations) identifies significant strong relation between SIGMAR1 Pro2 mutation and absence of response as measured both using MMSE and ADCS-ADL.Table 19.104108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center105108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center

[0228] Table 20 is a representation of one association rule showing the presence of Pro2 mutation in SIGMA1R implies a Low delta of MMSE with details on the rule metrics, extracted from data shown in Table 18).Table 20.

[0229] The mutation of SIGMAR1 Gin (Glutamine) 2 changed into Pro (Proline) 2 (SIGMAR1_p. Q2P) may have structural implication for the 3D conformation of the 3D structure of the SIGMAR1 protein which in turn may alter its functions and / or interactions. The crystal structure of the SIGMAR1 shows a trimer. Each monomer displays an N terminal trans-membrane alpha helix. The N terminus of the protein is thought to face the Cytosol and is believed to interact with other ligands. The Pro2 alteration may change the conformation of the N terminus of the helix by extending it, consistent with some alteration in function.

[0230] It is possible that the Gin (Q) to Pro (P) alteration impacts the length of the side chain (extended in Gin) as well as the ability of the amino acid to form an alpha helix. Proline is considered as a “helix breaker” which, in some instances, has been attributed to unwinding of a helix.

[0231] KEM® (version 3.6.2.) relations linking presence of the SIGMAR1 Pro2 mutation with cognitive and functional endpoints are shown in Table 19. Cognitive and functional measures are both clinical endpoints in regulatory approval for therapeutics in mild and mild-to-moderate Alzheimer's disease and directed to be significantly better than placebo as primary endpoints.106108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center

[0232] Given that 57-week Phase 2a ANAVEX2-73-002 study did not include a placebo arm, the KEM® analysis measured the two endpoints consisting of cognition (MMSE) and function (ADCS-ADL) with the following two methods: Each patient's own delta of MMSE and ADCS-ADL score (delta between Week 57 and Baseline).

[0233] Importantly, since Alzheimer’s disease is understood to be a progressing degenerative disease, an increase of MMSE (positive delta) and ADCS-ADL (positive delta) during 57 weeks in mild-to-moderate Alzheimer's disease is a rare positive event. Significantly, 100% of all patients which did improve MMSE (delta, n=9) and ADCS-ADL (delta, n=8) had no SIGMAR1 mutation. Further, 100% of patients bearing the SIGMAR1 Pro2 (n=5) mutation did not improve MMSE or ADCS-ADL delta. Available genetic data from all (100%) patients (n=21).

[0234] Similar results were obtained for the COMT variant COMT_p. L146fs (rs113895332 / rs61143203). 100% of all patients which did exhibit improved MMSE (both delta and slope) and ADCS-ADL (both delta and slope) (n=6) did not have the COMT_p. L146fs mutation. Further, 100% of patients bearing the COMT_p. L146fs (n=5) mutation did not improve MMSE or ADCS-ADL slope. The COMT_p. L146fs mutation is also associated with low delta MMSE and ADCS-ADL. Available genetic data from all (100%) patients (n=21).

[0235] As such, establishing the presence / absence of these mutations can be useful in treatment decisions regarding SIGMAR1 agonist treatment. The absence of these mutations is a diagnostic for enhanced SIGMAR1 agonist therapy such as A2-73 therapy. Establishing the presence of either or both of these mutations is a diagnostic for using other therapeutics as first-line treatment for neurodegenerative disorders.107108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center Table 21 presents the p-values results of the Mann-Whitney U test (also named Mann-Whitney-Wilcoxon test or MWWilc) between SIGMAR1 Pro2 and delta endpoints from Week 57 to baseline of MMSE and ADCS-ADL.Table 21.

[0236] SIGMAR1 Pro2 mutation leads to a significant difference in the numerical delta values of ADCS-ADL and MMSE delta (FIGS. 8C and 8F). The MWWilc test evaluates whether there is a significant difference in the medians of a continuous variable in two populations ( / .e. the two populations here: SIGMAR1 Pro2 mutation vs SIGMAR1 Gln2 mutation). The absence of SIGMAR1 Pro2 mutation is strongly linked to high ADCS-ADL Delta from BL (improvement). Similar results were found for the COMT Leu146 mutation (FIGS. 8A and 8D) and KANSL1 Pro304 / 946 / 1010 mutation (FIGS. 8B and 8E).

[0237] As noted above, 27 RNA expressions were associated with an improved or worsened response at week 57 versus baseline. Further selecting non CYP proteins and only high levels of RNA expression (excluding low and medium expression levels), shows that high expression levels of DHCR7, GAMT, IL10, KANSL1, MTUS1, PPARG and SIGMAR1 are associated with improved ADCS-ADL or MMSE Delta at week 57 versus baseline (Table 22).Table 22108108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center

[0238] Focusing on the intersection between the DNA and RNA analysis and on the subset of genes that were identified through the DNA analysis and affecting outcome as measured by both MMSE and ADCS-ADL, Table 23 shows higher RNA expression of 3 gene markers (SIGMAR1; COMT; KANSL1) associated with response as measured by MMSE or ADCS-ADL.Table 23109108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center

[0239] Notably, high RNA expression of SIGMAR1 is linked to high ANAVEX2-73 response (improvement) as shown in Table 24 and FIG. 9.Table 24110108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center100% of patients with low SIGMAR1 RNA expression did not improve in ADCS-ADL slope. Available genetic data from all (100%) patients. (N=21). Lift > 1.2; Conf. > 50%; Support > 3

[0240] Both DNA and RNA analysis provide convergent evidence as to the role of SIGMAR1, KANSL1 and COMT as predictors of outcome of SIGMAR1 therapy at week 57 as measured by a variation of MMSE and ADCS-ADL.

[0241] Table 25 shows a systematic analysis of the CYP genomic data (111 gene panel, from Table 15) identifies 11 SNPs significantly linked with outcome as measured by delta MMSE or delta ADCS-ADL at week 57. Correlations with improvement of MMSE and / or ADCS-ADL scores are the inclusion of variants CYP4V2, CYP27B1, CYP26C1. Correlations with an exclusion based on decline of MMSE and / or ADCS-ADL scores are the variants CYP39A1, CYP2D6, CYP2C8. Table 25111108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center112108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent CenterRules: n>3 patients, Conf. > 0.5, Lift >= 1.2, p-Value (Fisher Test or MWWilc.) < 0.05

[0242] Table 26 shows RNA expression of CYP4F2, CYP26A1 and CYP2D6 associated with response as measured by MMSE or ADCS-ADL.Table 26113108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent CenterRules: n>3 patients, Conf. > 0.5, Lift >=1.2, p-Value (Fisher Test or MWWilc.) < 0.05Example 5: ANAVEX®2-73-AD-004- Clinical Trial (NCT03790709)

[0243] ANAVEX®2-73-AD-004 was a randomized, double-blind, multicenter, placebo-controlled, 48-week trial with prespecified gene or GWAS identified genetic variant populations related to the mechanism of the pharmacological intervention. It is a 509 patient Phase 2b / 3 study (using mid or high oral dose of ANAVEX®2-73 or placebo), for the treatment of mild cognitive impairment (MCI) due to AD and mild AD (collectively known as “early AD”).

[0244] OBJECTIVE: Providing evidence of improved Precision Medicine neurology clinical treatment responses with blarcamesine for up to over 70% AD patients within prespecified and GWAS identified SIGMAR1 (ABCLEAR1), COL24A1 (ABCLEAR2) and SIGMAR1 / COL24A1 (ABCLEAR3) excluded missense genes populations with once-daily, oral therapeutic intervention in Early AD with a differentiated upstream and constitutional mechanism of action by enhancing autophagy through SIGMAR1 activation and restoration of cellular homeostasis.

[0245] SETTING: Multicenter - 52 medical research centers / hospitals in 5 countries.

[0246] INTERVENTION: 508 participants with early AD (Stage 3) were randomized to receive blarcamesine (n = 338) either in medium dose group 30 mg or in high dose group 50 mg or placebo (n = 170) oral capsules once daily for 48 weeks. The open-label-extension study ATTENTION-AD, continued for up to 192 weeks and demonstrated further robust clinical benefit with acceptable safety profile.

[0247] MAIN OUTCOME AND MEASURES: Further improved clinical and biomarker outcomes of ABCLEAR2 and ABCLEAR3 populations on top of the previously reported intent-to-treat (ITT) and ABCLEAR1 population. The co-primary cognitive and functional outcomes were assessed as change in ADAS-Cog13 and114108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center ADCS-ADL from baseline to 48 weeks. The outcomes include the secondary outcome CDR-SB as well as patient assessed clinical outcomes CGI-I, NPI-Q and QoL-AD as well as biomarkers of global brain volume changes measured by MRI. All clinical endpoints were analyzed using mixed model for repeated measures(MM RM), and volumetric MRI scans were analyzed by general linear model.

[0248] The following selection criteria was used to register patients:Inclusion Criteria:

[0249] Previous completion of participation in the ANAVEX2-73-AD-004 double-blind study.

[0250] Participants may be either outpatients, or residents of an assisted-living facility.

[0251] Participants must have a designated study partner, who spends at least 10hrs per week with the participant, in order that assessments eg. carer burden instruments are completed with true knowledge of the participant.

[0252] No suicidal ideation of type 4 or 5 in the Columbia Suicide Severity Rating Scale (C-SSRS) in the past 3 months ( / .e. active suicidal thought(s) with intent but without specific plan, or active suicidal thought(s) with plan and intent) OR suicidal behavior in the past 2 years ( / .e. actual attempt, interrupted attempt, aborted attempt, or preparatory acts or behavior).

[0253] Confirmation from the participant that, if of childbearing potential is not pregnant through urine pregnancy testing.Exclusion Criteria:

[0254] Adverse events (AEs) from the previous study (ANAVEX2-73-AD-004) that have not resolved, are moderate or severe, judged to be possibly related or related to study drug, and considered by the investigator to be a contraindication to extension study participation

[0255] Any condition or laboratory abnormality that would make the subject, in the judgment of the investigator, unsuitable for the study115108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center

[0256] Significant history of drug addiction (with the exception of nicotine dependence) or abuse (including alcohol, as defined in DSM-V or in the opinion of the investigator) within the last two years prior to informed consent, or a positive urine drug screen for cocaine, opioid, phencyclidine (PCP), amphetamine or marijuana at screening. Prescription medication yielding a positive drug screen are acceptable except for tricyclic antidepressants (e.g. Amitriptyline, Amoxapine, Desipramine, (Norpramin) Doxepin, Imipramine (Tofranil), Nortriptyline (Pamelor), Protriptyline (Vivactil), Trimipramine (Surmontil)).

[0257] Any known hypersensitivity to any of the excipients contained in the study drug formulation.Observations And Results:

[0258] The ITT population (mean age, 73.7 years; 225 [48.7%] women), consisted of 462 randomized participants, with ABCLEAR1 population comprising of 300 participants, ABCLEAR2 population of 336 participants and ABCLEAR3 of 222 participants. In the ABCLEAR3 population the co-primary outcomes as well all other clinical outcomes were statistically significant with blarcamesine group vs. placebo at Week 48 (ADAS-Cog13 difference of -4.179 [95% Cl -6.512, -1.845]; P=0.0005; ADCS-ADL difference of +3.131 [95%CI 0.720, 5.542]; P=0.0111; CDR-SB difference of -1.076 [95% Cl -1.645, -0.508]; P=0.0002). The clinical outcomes were strongest in the blarcamesine 30 mg cohort (ADAS-Cog13 difference of -4.739 [95% Cl -7.370, -2.108]; P=0.0004; ADCS-ADL difference of +4.245 [95%CI 1.518, 6.972]; P=0.0024; CDR-SB difference of -1.414 [95% Cl -2.054, -0.775]; P<0.0001). Whole brain volume loss blarcamesine group vs. placebo was significantly further decreased from ITT population (P=0.002) to ABCLEAR 3.0 population (P=0.001). Participants in the 30 mg group ABCLEAR3 full safety population with >1 serious treatment-emergent adverse events (TEAEs) occurred in 10 participants (12.7%) in the blarcamesine and 6 (9.1%) in the placebo group. Common TEAEs included dizziness, which was transient and mostly mild to moderate in severity. There were no deaths in the blarcamesine group and 1 in the placebo group.

[0259] CONCLUSIONS AND RELEVANCE: Blarcamesine, demonstrating a balanced safety profile with no associated neuroimaging adverse events, in both the ABCLEAR2 and ABCLEAR3 populations significant stabilization of clinical worsening 116108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center was demonstrated. Blarcamesines group vs. placebo response was 75.9% in the blarcamesine group at 48 weeks and was 84.7% in the 30 mg group, respectively on the prespecified primary cognitive endpoint ADAS-Cog13 for ABCLEAR3 population. Blarcamesine demonstrated consistently significant improved clinical effect for all clinical endpoints, which was in accordance with significant and further reduced brain atrophy. Furthermore, a significant absolute improvement in Quality of Life (QoL-AD) scores indicating a reversal of negative trajectory for Alzheimer's disease patients from baseline to end of trial was observed. The Phase llb / lll ANAVEX2-73-AD-004 clinical study confirmed blarcamesine consistent efficacy evidenced by the Precision Medicine paradigm leading to improved cognitive control and stabilization, including in a pre-specified ABCLEAR1 population and demonstrated consistent significant improvement for all clinical endpoints in the ABCLEAR2 and ABCLEAR3 populations, respectively. Coupled with a convenient once daily, oral pill administration, blarcamesine could represent a novel treatment in all early AD patients as well as for further improved outcome and patient benefit using Precision Medicine on top of standard of care which could alleviate significant medical and economic burden.

[0260] ANAVEX®2-73 met the primary endpoints of Alzheimer Disease Assessment Scale-Cognition (ADAS-Cog) and Activities of Daily Living Scale (ADCS-ADL). The trial results also met a key secondary endpoint Clinical Dementia Rating scale Sum of Boxes (CDR-SB) with statistically significant results. Further analysis split the testing population into APOE4-carrier and APOE4-Noncarriers. Tables 27 and 28 below showed p-values were significant for both CDR-SB and Clinical Global Impression-Improvement Scale (CGI-I), but only in APOE4-carrier patients. These results demonstrated that blarcamesine was more efficacious or effective in APOE4-carriers than in APOE4-Noncarriers.Table 27: Treatment Effects in APOE4 Carriers117108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent CenterTable 28: Treatment Effects in AP0E4 Noncarriers

[0261] These results demonstrated the differential effect of blarcamesine in AD patients carrying the APOE4 gene. Since the severity of Alzheimer's disease (AD) and age of its onset is highly correlated with the APOE4 gene, early detection of the gene in an individual allows early treatment with blarcamesine to delay early onset of AD or delay AD progression in individuals with early stage of AD.

[0262] Furthermore, patient populations possessing wild-type ( / .e., nonpolymorph versions) of the SIGMAR1 and / or COL24A1 genes were also studied. FIGS. 12 to 19 (and sub-figures) demonstrate that patient populations possessing wild-type ( / .e., non-polymorph versions) of the SIGMAR1 and / or COL24A1 genes experience increased efficacy with A2-73 therapy. This increased efficacy is demonstrated by MMRM analysis of change in one or more (or all) of ADAS-Cog13; ADCS-ADL; CDR-SB; CGI-Improvement; MMSE; NPI-Q; Patient QoL-AD; and Caregiver Qol-AD total score from baseline for the total (m)ITT population, or subgroups selecting for GWAS-identified gene variant populations.118108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center Example 6: COL24A1 gene impacts effectiveness of autophagy-enhancing blarcamesine in early stage and early onset Alzheimer’s disease

[0263] In our recent ANAVEX2-73-AD-004 Phase llb / lll clinical trial involving >500 patients with early AD, blarcamesine demonstrated efficacy and favorable safety, significantly slowing cognitive decline (ADAS-Cog13) and cognitive and functional impairment (CDR-SB) over 48 weeks compared to placebo in both the ITT and SIGMAR1 WT (wildtype) (ABCLEAR1; ABCLEAR: Alzheimer’s Blarcamesine Cognition - Longterm Effect and Resilience) populations, respectively. Benefits were observed irrespective of ApoE4 status and without the vascular safety concerns associated with injectable immunotherapies. Crucially, prespecified pharmacogenetic analysis confirmed target engagement, showing a more pronounced treatment effect in the ABCLEAR1 population, i.e., in participants of SIGMAR1 WT genotype (global population prevalence -70%), thus validating the fidelity of blarcamesine’s mechanism of action.

[0264] Extending clinical trial AD-004’s precision medicine findings, we investigated additional genetic factors impacting blarcamesine’s therapeutic response. For this, a pharmacogenomics GWAS (PGx-GWAS) to identify genetic factors underpinning an enhanced blarcamesine response was conducted. Here we report a DNA marker in the COL24A1 gene pointing to increased effectiveness of blarcamesine for ADAS-Cog13, ADCS-ADL and CDR-SB outcomes forCOL24A1 WT carriers, whom we refer to as ABCLEAR2 population (global population prevalence -71%). COL24A1 encodes the a1 chain of fibrillar collagen XXIV, a documented component of the human brain’s ECM. It is expressed predominantly by neurons of the hippocampal formation and cerebral cortex, positioning it in the spatial context to support neural circuits critical for cognition and memory - and thus directly relevant to blarcamesine’s therapeutic action.

[0265] Methods

[0266] Pharmacogenomics GWAS of the Blarcamesine Response

[0267] We conducted a PGx-GWAS on data from the ANAVEX2-73-AD-004 clinical trial (NCT03790709), flow-chart of subjects / methods applied shown in FIG.20, subjects’ demographics in Table 29. The analysis included 326 participants (198119108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center blarcamesine-treated, 128 placebo) for the cognitive outcome (ADAS-Cog13) and 329 (198 blarcamesine-treated, 131 placebo) for the functional-cognitive outcome (CDR-SB), for the respective difference blarcamesine versus placebo at timepoint 48 weeks. Whole-exome-sequencing (WES) was conducted on Illumina NovaSeq6000, with alignment to GRCh38 and variant calling following GATK best practices.Table 29: Demographics of ANAVEX2-73-AD-004 trial participants subjected to GWAS for enhanced blarcamesine response over placebo for outcomes ADAS-Cog13 and CDR-SBNumbers with number in parenthesis denotes n and %, or mean value and standard deviation.

[0268] A gene-by-treatment interaction analysis was conducted using PLINK2. A linear regression model tested the interaction term (SNPxTreatment Group) for 120108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center each variant, with the inverse normal-transformed change from baseline in ADAS-Cog13 or CDR-SB as the outcome. Models were adjusted for sex, ApoE4 carrier status, and the prespecified SIGMAR1 rs1800866 genotype. Genetic principal components were included to control for population stratification. Variants with a call rate <90% or deviation from Hardy-Weinberg equilibrium (p<1 O’5) were excluded. Genome-wide significance was set at p<5*10’8and second-level p<1 O’5.

[0269] Study Population and Phenotype Definition

[0270] We conducted a pharmacogenomic GWAS on data from the ANAVEX2-73-AD-004 clinical trial (NCT03790709), flow-chart of subjects / methods applied in FIG. 20, participants’ demographics in Table 29. The analysis cohort included trial participants with both whole-exome sequencing (WES) data and complete clinical outcome assessments at week 48. For the analysis of cognitive response, defined by the change from baseline in the Alzheimer’s Disease Assessment Scale-Cognitive Subscale (ADAS-Cog13), the cohort comprised 326 participants (198 blarcamesine, 128 placebo). For the cognitive-functional outcome of global function, assessed by the change in Clinical Dementia Rating-Sum of Boxes (CDR-SB), the cohort included 329 participants (198 blarcamesine, 131 placebo).

[0271] Given the non-normal distribution of the ADAS-Cog13 change scores, a rank-based inverse normal transformation (RINT) was applied prior to association testing.

[0272] Whole exome sequencing

[0273] DNA was extracted from venipunctured whole blood of AD-004 trial subjects using silica-gel based DNA extraction method followed by QC of the extracted DNA, then whole exome sequencing (WES) was performed as follows.

[0274] Paired-end sequencing was performed on an Illumina NovaSeq6000, generating FASTQ files for all samples. Raw data first underwent standard stringent sequencing QC, then were aligned to the human reference genome (GRCh38) using Burrows-Wheeler Aligner - Maximal Exact Matches (BWA-MEM) methodology. In a two-stage QC approach, the resulting Binary Alignment / Map (BAM) files then underwent a second alignment QC, which included sorting, duplicate marking, and121108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center base quality score recalibration (BQSR) using the Genome Analysis Toolkit (GATK) best practices pipeline, thus advancing high-quality, accurately aligned sequences to variant calling. No imputation methods were applied given the high quality and depth of sequences obtained, e.g. an average across-chromosome coverage of 70x.

[0275] Genotyping, Quality Control, and Statistical Analysis

[0276] WES sequences were subjected to the 2x QC format as outlined above, and a Variant Call Format (VCF) file was generated. Variants with a call rate <90% or a significant deviation from Hardy-Weinberg equilibrium (p <1 O’5) were excluded. A gene-by-environment (GxE) interaction analysis was performed using PLINK2 software, where the “environment” was the treatment assignment (blarcamesine vs. placebo). A linear fitting regression model for each variant was applied as follows:Phenotype = β0+ β1(SNP) + β2(Group) + β3(SNP x Group) + Covariates

[0277] The phenotype was the continuous change in ADAS-Cog13 (RINT-transformed) or CDR-SB. The model was additive for the SNP effect. Covariates included sex, APOE-E4 allele carrier status, and the SIGMAR1 rs1800866 genotype, a preselected genetic variant (encoding a loss-of-function missense of the molecular target of blarcamesine, the Sigma-1 receptor chaperone).

[0278] The interaction term (SNRxGroup) was the test of primary interest, identifying variants where the effect on clinical outcome was modified by treatment. Genome-wide significance was defined as p<5xl0“8. Due to the limited sample size inherent to our clinical trial-based pharmacogenomics, variants with suggestive significance (p<10’5) were also explored. A minor allele frequency (MAF) filter of <10% was applied post-analysis to generate the final dataset.

[0279] Genetic principal components were included as covariates to control for population stratification. This WES dataset exhibited minimal population stratification.

[0280] Analysis, QC and RINT were conducted with software PLINKv2.0, filtering of raw output and data visualization with J MP Pro.122108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center

[0281] Human brain gene expression data for COL24A1 were downloaded from the Allen Brain Atlas and processed with GraphPad Prism software.

[0282] Results

[0283] For our PGx-GWAS, Manhattan plots for association for blarcamesine treatment response in CDR-SB are shown in FIG. 21 A and for ADAS-Cog13 in FIG.21 B. The analysis yielded no locus with p<5x1 O’8. For p< 10’5, there were 8 genes with 11 loci for ADAS-Cog13, and 5 genes with 5 loci for CDR-SB (Table 30, Table 31)123108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent CenterTable 30: SNPs with at least one GWAS -logio[P]>5124108513585.1PATENT Attorney Docket No.: 085102-875342Via Patent CenterGenes: find more detail in Table. Effect size provided as absolute number.Ranking: missense principally ranking above intronic and other variant SNPs, primarily considering sum of -logio[P], secondarily sum of effect size.Table 31125108513585.1PATENT Attorney Docket No.: 085102-875342Via Patent Center126108513585.1PATENT Attorney Docket No.: 085102-875342Via Patent Center127108513585.1PATENT Attorney Docket No.: 085102-875342Via Patent Center

[0284] No SNP showed significance p<10-5for either. Fora criterion of one p<1×10-5and one p< 10'3, one SNP matched, namely rs60891279, located within the coding region of COL24A1, predicted change of arginine at position 641 to histidine (R641 H) in the helical domain of fibrillar collagen XXIVal. In order to rank SNPs, we generated an index: For chromosomal location and predicted impact of the SNP we accounted for whether the SNP originated from coding region orintronic (higher value for coding region), for coding region mis-sense was considered more valuable than synonymous, following a well-established prioritization approach in translational human genetics). Then we took into account combined -Iog10[p] values for ADAS-Cog13 and CDR-SB, with one required -logio[p]>5, additionally sum of absolute value of p3 effect size for each trial outcome metric. Applying this ranking, SNP rs60891279 was the unambiguous first rank, SNP rs184418609 was second-ranked. It is located in ADRA1D (adrenoceptor-a1D, S31G missense), and SNP rs17845226 was third-ranked, located in ANXA2 (annexin-A2, V116L(isoform1), V98L(isoform2) missense). Table 30 lists all SNPs with one -logio[p]>5, and Table 31 provides additional relevant background on each SNP, their respective allele frequencies, genes and encoded proteins, relationship to Alzheimer's disease, to neurodegenerative disease, known role in cell biological functioning and pathobiology, including autophagy and Sigmal -receptor chaperone function.

[0285] Taken together, we identified a missense SNP in COL24A1 for both clinical outcomes, ADAS-Cog13 and CDR-SB, as shared genetic marker common to both GWAS analyses, thus increasing confidence over SNPs only associated with one outcome.

[0286] The finding of SNP rs60892379 centers on amino acid position 641, within the collagen XXIVal protein encoded by the COL24A1 gene. Position 641 is in the helical subdomain of this fibrillar collagen protein, at position “Y” of the 31st repeat domain of amino acid triplet

[0287] G[glycine]-X-Y, of 309 such repeats, a general structural feature which is unique to collagens. This specific R641 H substitution is thus predicted to significantly alter the triple-helical collagen protein structure, and thus biological and biophysical properties of collagen XXIV.128108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center

[0288] Individuals carrying the wild type (WT) version of COL24A1, encoding arginine at position 641 (global population prevalence -71%), exhibited a significantly more effective response to blarcamesine compared to those with the missense mutation encoding histidine (R641H).

[0289] To recapitulate, this genetically encoded divergence enabled a precision medicine paradigm through three defined populations, each with statistically-significantly improved clinical outcomes: The clinical study provided evidence of improved treatment responses with blarcamesine for >70% AD patients within pre-specified SIGMAR1 WT (ABCLEAR1) and GWAS-identified COL24A1 WT (ABCLEAR2). Their combined genotype, SIGMAR1 WT; COL24A1 WT (ABCLEAR3) still has a 50% population prevalence. All ABCLEAR genetically-defined populations excluded the respective missense genetic variants. Effect sizes in ABCLEAR2 and ABCLEAR3 were robustly increased over ITT and ABCLEAR1. All groups defined by COL24A1 WT (ABCLEAR2, ABCLEAR3) showed statistically significant differences blarcamesine vs placebo at 48 weeks for all outcome criteria, for example ADAS-Cog13 -3.901 (ABCLEAR2), -4.179 (ABCLEAR3) (both p<0.001), ADCS-ADL 3.538 (ABCLEAR2, p<0.001), 3.131 (ABCLEAR3, p=0.0011), CDR-SB -0.986 (ABCLEAR2), -1.076 (ABCLEAR3) (both p<0.001), interestingly also for attenuation of grey matter whole brain atrophy: 68.8% slowing (ABCLEAR2, p=0.002), 65.2% slowing (ABCLEAR3, p=0.013).

[0290] Expression in critical brain regions by neurons: COL24A1 is expressed in brain areas fundamental for cognition, memory and cognitive-executive functioning, notably the hippocampal formation (including CA1, its stratum radiatum and subiculum) and the cortex. This provides the critical spatial context for blarcamesine to enhance autophagy and exert its neuroprotective effects.

[0291] In terms of cellular lineage, as revealed by publicly available brain atlases and a recent landmark publication, COL24A1 is dominantly expressed by neurons, with minor expression by glia and pericytes of the blood-brain-barrier.These expression data were derived from normal control adult brains, and together with the fact that collagen XXIVcH is a fibrillar collagen, point toward its physiologic function in the human brain in the ECM compartment of regions highly relevant for memory, cognition and cognitive-executive function.129108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center

[0292] In addition, we analyzed catalogued human brain COL24A1 gene expression data of aging controls vs a dementia cohort. We found significantly upregulated expression in hippocampus over cortex (2 cortical areas sampled, temporal and parietal), and significantly elevated expression in cortex over white matter (FIG. 22). There were no statistical differences between control and dementia cohort. There were no differences within the dementia group between more severe dementia and all dementia cases, using Braak and Cerad indices as indicators for severity. These data extend our understanding: aging subjects show robust COL24A1 expression in memory- and cognition-critical areas, with age-associated preservation of strong expression in hippocampal neurons, and unrelenting expression in dementia.

[0293] Discussion

[0294] We present here the pharmacogenomics GWAS of clinical trial ANAVEX2-73-AD-004, querying for an enhanced blarcamesine response which allowed us to examine 324 / 327 patients by genetic analysis for their treatment response to blarcamesine vs placebo at 48 weeks, for cognitive outcome ADAS-Cog13 as one measurable (324 pts), and cognitive-functional outcome CDR-SB as another measurable (327 pts). The one SNP DNA polymorphism we identified for both metrics was SNP rs60891279, located in COL24A1, a missense mutation predicted to encode an amino acid substitution R641 H in fibrillar collagen XXIVal, altering its triple helical structure. The WT allele of COL24A1, present in 71% of populations, showed a robustly enhanced blarcamesine response.

[0295] Strong positive effect on top of the impact of an already effective antiAlzheimer therapeutic by a widely prevalent genetic polymorphism is novel. Our quest identified COL24A1, encoding a fibrillar collagen with a previously sparse record of CNS involvement, expressed in neurons of hippocampal formation and cortex, indicating key relevance for cognition, memory and cognitive-executive function. The impact of missense mutation R641H in collagen XXIVal on treatment response to autophagy-enhancing blarcamesine could be via mis-sculpting of the ECM and / or effects of the R641 H substitution via intracellular procollagen XXIV unfavorably pre-loading the lysosomal autophagy machinery.130108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center

[0296] Our pharmacogenomics finding of COL24A1 as a blarcamesine response gene, derived from 324 / 327 clinical trial patients mandates repeat in an independent cohort, yet the following features of our investigation elevate confidence: (i) significant association of the identified COL24A1 SNP with both outcome measures, ADAS-Cog13 and CDR-SB, despite the modest cohort size; (ii) our trial population originated diversely from 52 clinical centers and 5 countries; (iii) brain expression patterns of COL24A1 in physiology, aging and dementia strongly support a previously underrecognized role for this gene, and its product, fibrillar collagen XXI Va1, in memory and cognitive-executive function; (iv) recent COL24A1-related research findings further support and significantly extend this argument. These COL24A1 -related findings indicate regulated expression in a relevant region of the brain in an AD context. Recent Chinese human genetic finding highlights that Han Chinese genetic background combined with 6kB deletion in COL24A1 loss-of-function caused hereditary Parkinson’s disease, a neurodegenerative disorder of more select neuronal lineages that bears a strong pathobiological connection to autophagy-mitophagy, indicating COL24A1’s role in autophagy via human genetic evidence.

[0297] As a result of the COL24A1 rs60891279 missense mutation, the substitution of arginine (R) with histidine (H) at position 641 represents a subtle but potentially significant structural change in the collagen XXIVol protein. It is a fibrillar collagen and as such a newly recognized component of the brain's ECM - the intricate scaffold providing structural and trophic support to neurons, glial cells and the CNS’ vasculature. Position 641 is in the Y position of the characteristic collagen amino acid triplet, G[lycine]-X-Y, where in all collagens arginine is well-represented at 11.4%, and known to contribute to stability and rigidity of the mature collagen triple-helix in the ECM by forming salt bridges with lysine. An R^H substitution in a fibrillar collagen as a direct cause for a hereditary collagen disorder with a developmental clinical phenotype such as osteogenesis imperfecta has not been reported. However, a subtle phenotype of COL1A1(R618H) was described in a mother and more accentuated in her son, both heterozygotes, with mild weakness of connective tissues and short stature. There are multiple reports of R^H polymorphisms in helical segments of COL1A1 and COL1A2 in osteogenesis imperfecta, add-on to “classic” glycine mutations, which are discussed as modifiers.131108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center

[0298] There is an existing case for an ECM protein harboring a diseasecausing R^H substitution and linked to autophagy, namely granular corneal dystrophy, an autosomal-dominant anterior eye disorder caused by substitution R124H in the TGFp-induced protein TGFBIp. This corneal keratocyte ECM protein is degraded by autophagy during biosynthesis, yet TGFBIp(R124H) accumulates in corneal fibroblasts of patients with granular corneal dystrophy, thus suggesting causality for the dystrophic process affecting their corneas, which was rescuable in explanted cells by targeted overexpression of autophagy master-regulator, TFEB.

[0299] The mechanism by which the COL24A1 rs60891279 missense mutation (R641 H) attenuates treatment response appears to involve two non-mutually exclusive pathways, centered on the ECM’s role in proteostasis. Thus the emerging concept how collagen XXIVa1(R641H) makes patients respond less effectively is (i) an intracellular-intraneuronal preload of autophagy machinery caused by increased proteostatic stress via triple-helical pro-collagen XXIVa1(R641H) that is occupying to capacity the pre-secretion cellular qualitycontrol system, and (ii) by its modulatory effects on the ECM environment of memory- and cognition-critical areas of the brain where blarcamesine acts to enhance autophagy.

[0300] We therefore propose several rationally based concepts consistent with COL24A1's localized expression and blarcamesine's known mechanism-of-action as autophagy enhancer, schematic depicted in FIG. 23.

[0301] First, we propose an intracellular autophagy preload mechanism. The R641 H substitution, located in the triple-helical domain, is predicted to disrupt critical arginine-lysine salt bridges, compromising procollagen stability. Given that 10-30% of procollagens are typically degraded intracellularly before secretion, a misfolded variant could place a disproportionate burden on the neuronal autophagy-lysosome system. This constant, low-level proteostatic stress would preoccupy the very clearance machinery that blarcamesine is designed to enhance. Consequently, when treatment is initiated, the autophagy response in R641H carriers is blunted, as their system is already operating near capacity. The absence of this variant as an AD risk allele in large-scale GWAS suggests this burden is clinically silent until the system is pharmacologically challenged.132108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center

[0302] Second, the mutation likely alters the ECM microenvironment in cognition-critical regions like the hippocampal formation. The defective collagen XXIVcd could integrate into the ECM, changing local tissue properties such as mechanical stiffness or ionic gradients. This altered milieu may impair SIGMAR1 activation by blarcamesine or the efficiency of downstream autophagic execution. This concept is supported by the localized expression of COL24A1 and the increasingly recognized role of the ECM in bidirectionally regulating autophagy in neurodegeneration. An attractive, though provisional, hypothesis is that collagen XXIVcd functions within or alongside perineuronal nets (PNNs). These specialized ECM structures balance synaptic stability and plasticity; a compromised PNN environment due to the R641H mutation could directly hinder the synaptic resilience promoted by autophagy enhancement.

[0303] Furthermore, collagen XXIV may act synergistically with other ECM components. It has been suggested to interact with collagen V, whose a2 subunit is downregulated in the AD hippocampus formation. Afunctional deficit in one collagen partner (collagen XXIVcd (R641H)) coupled with reduced expression of the other (collagen V) could critically undermine the structural and signaling scaffold necessary for optimal autophagy enhancement. This network connects to key signaling pathways, as collagen V tethers to integrins, which are known to interact with SIGMAR1, providing a potential direct link between ECM integrity and the drug’s mechanism of action.

[0304] This discovery has immediate clinical relevance. Blarcamesine has already demonstrated a favorable safety profile and significant benefit in a broad early AD population. The identification by PGx-GWAS of the COL24A1 WT genotype allows for a precision medicine strategy (ABCLEAR2) that can be readily implemented to optimize outcomes for a majority of patients. This approach is further refined by combining it with the previously identified, prespecified SIGMAR1 WT genotype (ABCLEAR1), creating a highly responsive ABCLEAR3 population (SIGMAR1 WT; COL24A1 WT). This represents a significant advance over current parenteral immunotherapies, offering a convenient oral option with a genetically defined, high-prevalence responder profile.133108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center

[0305] Mechanistically, our findings point to the CNS ECM as a critical modulator of therapeutic response in neurodegeneration. Our discovery establishes a novel paradigm where common genetic variation in a structural ECM protein can profoundly influence drug efficacy by interacting with a specific mechanism of action, in this case, enhanced autophagy by SIGMAR1 activation.Example 1. o-1 Agonist Responder / Non-Responder Determination

[0306] Twelve subjects diagnosed with Alzheimer’s disease, aged 57 to 71, are tested for the Q2P polymorphism in the SIGMAR1 gene. It is determined that two subjects carry the amino acid proline at the 2 position of the SIGMAR1 protein. It is further determined that the remaining 10 subjects carry a glutamine at that position on the receptor protein. The group with amino acid proline at the 2 position of the SIGMAR1 protein is determined unsuitable for Sigma-1 agonist therapy. The group with amino acid glutamine at the 2 position of the SIGMAR1 protein is determined suitable for Sigma-1 agonist therapy.Example 8. u-1 Agonist Responder Determination and Therapy

[0307] A 74 year old male presents with diagnosed Alzheimer’s disease. His SIGMAR1 gene is analyzed to the extent that it is determined that he carries glutamine at the 2 position of the SIGMAR1 gene. This is the wildtype. He is treated with A2-73 daily with a 30 mg oral dose for 6 months. His disease is seen to be stable and not progressed over that period.Example 9. o-1 Agonist Responder Determination and Therapy

[0308] A 74 year old male presents with diagnosed Alzheimer’s disease. A cheek swab is taken and a genotype assay is performed to determine the presence of the SIGMAR1 2P genotype. By such analysis it is determined that the subject carries glutamine at the 2 position of the SIGMAR1 gene receptor (protein). He is treated with ANAVEX2-73 daily with a 30 mg oral dose. For 6 months. His disease is seen to be stable and not progressed over that period.Example 10. o-1 Agonist Responder Determination and Therapy

[0309] A 66 year old female presents with diagnosed dementia. Her SIGMAR1 gene is analyzed to the extent that it is determined that she carries proline134108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center at the 2 position of the SIGMAR1 gene. She is declined treatment with a sigma-1 agonist. She is referred for other therapeutic treatment options.Example 11. a-1 Agonist Responder Determination and Therapy

[0310] A 60 year old female presents with diagnosed dementia. Her SIGMAR1 gene is analyzed to the extent that it is determined that she carries glutamine at the 2 position of the SIGMAR1 gene. She is treated with ANAVEX2-73 daily with a 80 mg oral dose in combination with 1.5 mg oral rivastigmine twice per day for 6 months. Her disease is seen to be stable and not progressed over that period.Example 12. u-1 Agonist Responder Determination and Therapy

[0311] A 64 year old male presents with diagnosed Alzheimer’s disease. His COMT gene is analyzed to the extent that it is determined that he does not carry COMT dbsnp ID rs113895332 / rs61143203 mutation. He is treated with the A2-73 daily with a 30 mg oral dose for 6 months. His disease is seen to be stable and not progressed over that period.Example 13. a-1 Agonist Responder Determination and Therapy

[0312] A 71 year old female presents with diagnosed Alzheimer’s disease. Her SIGMAR1 gene is analyzed to the extent that it is determined that she does not carry SIGMAR1 rs1800866. She is treated with A2-73 daily with a 30 mg oral dose for 6 months. Her disease is seen to be stable and not progressed over that period.Example 14. a-1 Agonist Responder Determination and Therapy

[0313] An 81 year old male presents with diagnosed dementia. It is determined that he carries the KANSL1_p. P1010L / P304L / P946L mutation. He is declined treatment with a sigma-1 agonist, and referred for other therapeutic treatment options.Example 15. a-1 Agonist Responder Determination and Therapy

[0314] An 78 year old male presents with diagnosed dementia. He carries glutamine at the 2 position of the SIGMAR1 gene. It is determined that he carries the KANSL1_p. P1010L / P304L / P946L mutation. He is treated with135108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center ANAVEX2-73 daily with a 70 mg oral dose in combination with 7 mg oral memantine daily for 6 months. His disease is seen to be stable and not progress over that period.Example 16. cr-1 Agonist Responder Determination and Therapy

[0315] A 74 year old male presents with diagnosed Alzheimer’s disease. His COL24A1 gene is analyzed to the extent that it is determined that he carries the wildtype version of that gene. He is treated with A2-73 in accordance with the above disclosure. His disease is seen to be stable and not progressed over 6 months. Example 17. o-1 Agonist Responder Determination and Therapy

[0316] A 66 year old female presents with diagnosed Alzheimer’s disease. Her COL24A1 gene is analyzed to the extent that it is determined that she carries the COL24A1 (rs60891279) polymorphism. She is declined treatment with a Sigma-1 agonist. She is referred for other therapeutic treatment options.Example 18. u-1 Agonist Responder Determination and Therapy

[0317] A 74 year old male presents with diagnosed Alzheimer’s disease. His SIGMAR1 gene is analyzed to the extent that it is determined that he carries glutamine at the 2 position of the SIGMAR1 gene and also carries the wild type version of COL24A1. He is treated with A2-73 in accordance with the above disclosure. His disease is seen to be stable and not progressed over 6 months. Example 19. o-1 Agonist Responder Determination and Therapy

[0318] A 60 year old male presents with diagnosed with early onset Alzheimer’s disease. His genes are analyzed to the extent that it is determined that he possesses the wild type version of COL24A1. He is treated with A2-73 in accordance with the above disclosure. His disease is seen to be stable and not progressed over 6 months.Example 20. o-1 Agonist Responder Determination and Therapy

[0319] A 69 year old female presents with diagnosed Alzheimer’s disease. Her genes are analyzed to the extent that it is determined that she does not carry the APOE4 gene. She is declined treatment with a Sigma-1 agonist. She is referred for other therapeutic treatment options.136108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center Example 21. u-1 Agonist Responder Determination and Therapy

[0320] A 64 year old male presents with early onset Alzheimer’s disease. His genes are analyzed to the extent that it is determined that he possesses the wild type version of COL24A1. He is treated with A2-73 in accordance with the above disclosure. His symptoms are seen to be stable and not progressed over that period.Example 22. o-1 Agonist Responder Determination and Therapy

[0321] A 71 year old male presents with mild / early stage Alzheimer’s disease. His genes are analyzed to the extent that it is determined that he possesses the wild type version of COL24A1. He is treated with A2-73 in accordance with the above disclosure. His symptoms are seen to be stable and not progressed over that period.Example 23. cr-1 Agonist Responder Determination and Therapy

[0322] A 73 year old male presents with mild / early stage Alzheimer’s disease. Her COL24A1 gene is analyzed to the extent that it is determined that he carries the COL24A1 (rs60891279) polymorphism. He is declined treatment with a Sigma-1 agonist and is referred for other therapeutic treatment options.Example 24. o-1 Agonist Responder Determination and Therapy

[0323] A 62 year old female presents with early onset Alzheimer’s disease. Her genes are analyzed to the extent that it is determined that she carries either one or both of ADRA1D (re184418609) and ANXA2 (rs17845226). She is declined treatment with a Sigma-1 agonist and is referred for other therapeutic treatment options.137108513585 1

Claims

PATENT Attorney Docket No.: 085102-875342Via Patent Center CLAIMS1. A method of selecting a subject responsive to Sigma-1 receptor therapy, the method comprising:(a) obtaining or having obtained the results of a test of a biological sample from the subject which determines the presence of at least one polymorphism selected from the group consisting of APOE4 (rs429358 and / or rs7412), COL24A1 (rs60891279), ADRA1 D (re184418609), ANXA2 (rs17845226), SIGMAR1_Q2P, COMT_L146fs, KANSL1_P1010L / P304L / P946L, DHCR7_M220T, HLA-DRB1_ A244T, HLA-DRB1_ S66Y, HLA-DRB1_ Y89S, MS4A6E_M59T, RIN3_ H215R, DPYD_I543V, and combinations thereof in the subject; and(b) excluding the subject as non-responsive to Sigma-1 receptor therapy if a polymorphism is present in the subject the subject.

2. A method of selecting subjects responsive to Sigma-1 receptor therapy, the method comprising:(a) detecting or having detected whether at least one polymorphism is present at a genetic locus in a biological sample obtained from the subject by sequencing the genetic locus comprising the polymorphism; and(b) excluding the subject as non-responsive to Sigma-1 receptor therapy if a polymorphism is present in the subject;wherein the at least one polymorphism is selected from the group consisting of APOE4 (rs429358 and / or rs7412), COL24A1 (rs60891279), ADRA1D(re184418609), ANXA2 (rs17845226), SIGMAR1_Q2P, COMT_L146fs, KANSL1_P1010L / P304L / P946L, DHCR7_M220T, HLA-DRB1_ A244T, HLA-DRB1_ S66Y, HLA-DRB1_ Y89S, MS4A6E_M59T, RIN3_ H215R, DPYD_I543V, and combinations thereof.

3. A method of selecting subjects responsive to Sigma-1 receptor therapy, the method comprising:(a) obtaining or having obtained the results of a test of a biological sample from the subject which determines the level of expression of RNA encoded by at 138108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center least one gene selected from the group consisting of APOE4, COL24A1, ADRA1D, ANXA2, SIGMAR1, COMT, KANSL1, DHCR7, HLA-DRB1, MS4A6E, RIN3, DPYD, and combinations thereof; and(b) comparing the level of the RNA in the test sample to the level of the RNA in a subject responsive to Sigma-1 receptor therapy;wherein a level of the RNA in the test sample substantially similar to the level of the RNA in the subject responsive to Sigma-1 receptor therapy identifies a subject responsive to Sigma-1 receptor therapy.

4. A method of selecting subjects responsive to Sigma-1 receptor therapy, the method comprising:(a) determining or having determined the level of expression of at least one RNA encoded by a gene selected from the group consisting of APOE4, COL24A1, ADRA1D, ANXA2, SIGMAR1, COMT, KANSL1, DHCR7, HLA-DRB1, MS4A6E, RIN3, DPYD, and combinations thereof; and(b) excluding the subject as non-responsive to Sigma-1 receptor therapy if the level of the RNA in the test sample is substantially different from the level of the RNA in a subject responsive to Sigma-1 receptor agonist therapy.

5. A method of treating a subject in need of Sigma-1 receptor therapy, the method comprising:(a) obtaining or having obtained the results of a test of a biological sample from the subject which detects the presence of at least one polymorphism selected from the group consisting of APOE4 (rs429358 and / or rs7412), COL24A1 (rs60891279), ADRA1 D (re184418609), ANXA2 (rs17845226), SIGMAR1_Q2P, COMT_L146fs, KANSL1_P1010L / P304L / P946L, DHCR7_M220T, HLA-DRB1_ A244T, HLA-DRB1_ S66Y, HLA-DRB1_ Y89S, MS4A6E_M59T, RIN3_ H215R, DPYD_I543V, and combinations thereof in the subject; and(b) administering a therapeutically effective amount of a Sigma-1 receptor agonist to the subject if a polymorphism is not present in the subject.139108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center 6. A method of treating a subject in need of Sigma-1 receptor therapy, the method comprising:(a) obtaining or having obtained the results of a test of a biological sample from the subject which determines the level of expression of RNA encoded by at least one gene selected from the group consisting of APOE4, COL24A1, ADRA1D, ANXA2, SIGMAR1, COMT, KANSL1, DHCR7, HLA-DRB1, MS4A6E, RIN3, DPYD, and combinations thereof;(b) comparing the level of the RNA in the biological sample to the level of the RNA in a subject responsive to Sigma-1 receptor agonist therapy; and (c) administering a therapeutically effective amount of a Sigma-1 receptor agonist to the subject if a level of the RNA in the biological sample is substantially similar to the level of the RNA in a subject responsive to Sigma-1 receptor therapy.

7. A method of detecting the presence of a polymorphism associated with response to Sigma-1 receptor therapy in a subject, the method comprising:(a) obtaining or having obtained a biological sample from the subject; and (b) detecting the polymorphism in the biological sample by evaluating the sequence of a genetic locus comprising the polymorphism;wherein the at least one polymorphism is selected from the group consisting of APOE4 (rs429358 and / or rs7412), COL24A1 (rs60891279), ADRA1D(re184418609), ANXA2 (rs17845226), SIGMAR1_Q2P, COMT_L146fs, KANSL1_P1010L / P304L / P946L, DHCR7_M220T, HLA-DRB1_ A244T, HLA-DRB1_ S66Y, HLA-DRB1_ Y89S, MS4A6E_M59T, RIN3_ H215R, DPYD_I543V, and combinations thereof in the subject.

8. The method of any one of claims 1, 2, 5, and 7, wherein the at least one polymorphism is selected from APOE4 (rs429358 and / or rs7412), COL24A1 (rs60891279), ADRA1D (re184418609), ANXA2 (rs17845226), SIGMAR1_Q2P, COMT_L146fs, KANSL1_P1010L / P304L / P946L, and combinations thereof.140108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center 9. The method any one of claims 1, 2, 5, and 7, wherein the polymorphism is selected from APOE4 (rs429358 and / or rs7412), COL24A1 (rs60891279), ADRA1D (re184418609), ANXA2 (rs17845226), SIGMAR 1_Q2P, and combinations thereof.

10. The method any one of claims 1, 2, 5, and 7, wherein the polymorphism is selected from APOE4 (rs429358 and / or rs7412), COL24A1 (rs60891279), ADRA1D (re184418609), ANXA2 (rs17845226) and combinations thereof.

11. The method any one of claims 1, 2, 5, and 7, wherein the polymorphism is selected from COL24A1 (rs60891279), ADRA1D (re184418609), ANXA2 (rs17845226and combinations thereof.

12. The method any one of claims 1, 2, 5, and 7, wherein the polymorphism is COL24A1 (rs60891279).

13. The method of any one of claims 3, 4, and 6, wherein the gene is selected from APOE4, COL24A1, ADRA1D, ANXA2, SIGMAR1, COMT, KANSL1, DHCR7, HLA-DRB1, MS4A6E, RIN3, DPYD, and combinations thereof.

14. The method of any one of claims 3, 4, and 6, wherein the gene is selected from APOE4, COL24A1, ADRA1D, ANXA2, SIGMAR1, and combinations thereof, or wherein the gene is COL24A1, or where the gene is APOE4, or where the gene is ADRA1 D, or where the gene is ANXA2, or where the gene is SIGMAR1.

15. The method of any one of claims 3, 4, and 6, wherein the gene is selected from APOE4, COL24A1, ADRA1D, ANXA2, and combinations thereof.

16. The method of any one of claims 3, 4, and 6, wherein the gene is selected from COL24A1, ADRA1D, ANXA2, and combinations thereof.

17. The method of any one of claims 3, 4, and 6, wherein the gene is COL24A1.141108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center 18. The method of any one of claims 5 and 6, further comprising administering to the subject an alternative to Sigma-1 receptor agonist therapy if at least one polymorphism is present in the subject.

19. The method of any one of claims 1-4 and 7-18, wherein the Sigma-1 receptor agonist therapy comprises administration of a Sigma-1 receptor agonist selected from:tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride (A2- 73);tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine (A2-73 freebase); tetrahydro-N, N-dimethyl-5,5-diphenyl-3-furanmethanamine hydrochloride (A1-41);tetrahydro-N, N-dimethyl-5,5-diphenyl-3-furanmethanamine (A1-41 freebase); 1-(2,2-diphenyltetrahydrofuran-3-yl)-N-methylmethanamine hydrochloride (A19-144);1-(2,2-diphenyltetrahydrofuran-3-yl)-N-methylmethanamine (A19-144 freebase);(1-(2,8-dimethyl-1-thia-3,8-diazaspiro(4.5)dec-3-yl)-3-(1H-indol-3-yl)propan-1-one) (A3-71);an enantiomer thereof, a crystal form thereof, a co-crystal form thereof, a pharmaceutically acceptable salt thereof, a crystal or co-crystal of the pharmaceutically acceptable salt thereof, and any combination thereof.

20. The method of any one of claims 1-4 and 7-18, wherein the Sigma-1 receptor agonist therapy comprises administration of a Sigma-1 receptor agonist selected from:tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine in amorphous form;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine in crystal Form I;142108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride in crystal Form I;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride in crystal Form II;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride in crystal Form III;(+) tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride in amorphous form;(-) tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride in amorphous form;(+) tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride in a crystal form;(-) tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride in a crystal form;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrogen fumarate in amorphous form;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrogen fumarate in crystal Form I;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrogen fumarate in crystal Form II;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrogen fumarate in crystal Form III;143108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrogen fumarate in crystal Form IV;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrogen fumarate in crystal Form V;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine mesylate in crystal Form I;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine sulfate in crystal Form I;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine sulfate in crystal Form II;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine oxalate in crystal Form I;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine oxalate in crystal Form II;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine dihydrogen phosphate in crystal Form I;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine edisylate in crystal Form I;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine benzoate in crystal Form I;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrobromide in crystal Form A;144108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrobromide in crystal Form B;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine maleate in crystal Form S5;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine maleate in crystal Form S6;co-crystal Form CSII of tetrahydro-N, N-dimethyl-2,2-diphenyl-3- furanmethanamine hydrochloride with tartaric acid;co-crystal Form CSIII of tetrahydro-N, N-dimethyl-2,2-diphenyl-3- furanmethanamine hydrochloride with citric acid;co-crystal Form CSIV of tetrahydro-N, N-dimethyl-2,2-diphenyl-3- furanmethanamine hydrochloride with malic acid;a co-crystal of tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride with zinc chloride;a co-crystal of (-) tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride with zinc chloride;a co-crystal of (+) tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride with zinc chloride; andany combination thereof.

21. The method of any one of claims 5, 6, and 8-18, wherein the Sigma-1 receptor agonist is selected from:tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride (A2- 73),tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine (A2-73 freebase),145108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Centertetrahydro-N, N-dimethyl-5,5-diphenyl-3-furanmethanamine hydrochloride (A1- 41);tetrahydro-N, N-dimethyl-5,5-diphenyl-3-furanmethanamine (A1-41 freebase); 1-(2,2-diphenyltetrahydrofuran-3-yl)-N-methylmethanamine hydrochloride (A19-144);1-(2,2-diphenyltetrahydrofuran-3-yl)-N-methylmethanamine (A19-144 freebase);(1-(2,8-dimethyl-1-thia-3,8-diazaspiro(4.5)dec-3-yl)-3-(1H-indol-3-yl)propan-1-one) (A3-71);an enantiomer thereof, a crystal form thereof, a co-crystal form thereof, a pharmaceutically acceptable salt thereof, a crystal or co-crystal of the pharmaceutically acceptable salt thereof, and any combination thereof.

22. The method of any one of claims 5, 6, and 8-18, wherein the Sigma-1 receptor agonist is selected from:tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine in amorphous form;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine in crystal Form I;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride in crystal Form I;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride in crystal Form II;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride in crystal Form III;(+) tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride in amorphous form;146108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center(-) tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride in amorphous form;(+) tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride in a crystal form;(-) tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride in a crystal form;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrogen fumarate in amorphous form;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrogen fumarate in crystal Form I;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrogen fumarate in crystal Form II;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrogen fumarate in crystal Form III;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrogen fumarate in crystal Form IV;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrogen fumarate in crystal Form V;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine mesylate in crystal Form I;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine sulfate in crystal Form I;147108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Centertetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine sulfate in crystal Form II;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine oxalate in crystal Form I;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine oxalate in crystal Form II;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine dihydrogen phosphate in crystal Form I;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine edisylate in crystal Form I;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine benzoate in crystal Form I;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrobromide in crystal Form A;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrobromide in crystal Form B;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine maleate in crystal Form S5;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine maleate in crystal Form S6;co-crystal Form CSII of tetrahydro-N, N-dimethyl-2,2-diphenyl-3- furanmethanamine hydrochloride with tartaric acid;148108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Centerco-crystal Form CSIII of tetrahydro-N, N-dimethyl-2,2-diphenyl-3- furanmethanamine hydrochloride with citric acid;co-crystal Form CSIV of tetrahydro-N, N-dimethyl-2,2-diphenyl-3- furanmethanamine hydrochloride with malic acid;a co-crystal of tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride with zinc chloride;a co-crystal of (-) tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride with zinc chloride;a co-crystal of (+) tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride with zinc chloride; andany combination thereof.

23. The method of any one of claims 5, 6, and 18-22, wherein the administering or administration comprises a route of administration selected from the group consisting of oral administration, subcutaneous injection, intravenous injection, intraocular injection, intradermal injection, intramuscular injection, intraperitoneal injection, intratracheal administration, inhalation, intranasal administration, sublingual administration, buccal administration, rectal administration, vaginal administration, and topical administration.

24. The method of any preceding claim, wherein the Sigma-1 receptor therapy and / or administration of a Sigma-1 receptor agonist comprises a treatment period of about 12 weeks or less.

25. The method of claim 24, wherein the treatment period is about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 9 weeks, about 10 weeks, about 11 weeks, about 12 weeks.149108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center 26. The method of any one of claims 24-25, wherein the Sigma-1 receptor agonist is administered daily.

27. The method of any one of claims 24-25, wherein the Sigma-1 receptor agonist is tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine in Crystal Form I.

28. The method of any one of claims 24-25, wherein the Sigma-1 receptor agonist is tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride in Crystal Form I.

29. The method of any one of claims 19-28, wherein the Sigma-1 receptor agonist is administered daily in an amount of about 5 mg to about 60 mg.

30. The method of any one of claims 19-28, wherein the Sigma-1 receptor agonist is administered in an oral liquid comprising up to about 30 mg of the Sigma-1 receptor agonist.

31. The method of any one of claims 19-26, wherein the Sigma-1 receptor agonist comprises tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine or tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride in an oral solution, and wherein the tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine or the tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride is administered once daily for a period of 12 weeks in an escalating dose regimen starting at about 5 mg to about 10 mg and ending at about 20 mg to about 30 mg.

32. The method of any preceding claim, wherein the subject has a condition selected from the group consisting of a neuroprotective condition, a neurodegenerative condition, and a neurodevelopmental condition.

33. The method of claim 32, wherein the neuroprotective condition, the neurodegenerative condition, or the neurodevelopmental condition comprises cognitive impairment, brain atrophy, Lewy body dementia, stroke, traumatic brain injury, infections, spinal cord injury, Alzheimer’s disease (AD) (including early onset150108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center AD), Parkinson’s disease, dementia, Huntington’s disease, Amyotrophic lateral sclerosis, Prion disease, Rett Syndrome, Fragile X Syndrome, cerebral palsy, Angelman syndrome, Williams syndrome, pervasive developmental disorder not otherwise specified (PDD-NOS), childhood disintegrative disorder, Smith-Magenis syndrome, multiple sclerosis, Fronto-temporal dementia, motor neuron disease (MND), spinocerebellar ataxia (SCA), spinal muscular atrophy (SMA), autism spectrum disorder, schizophrenia, post-traumatic stress disorder (PTSD), and any combination thereof.

34. The method of any preceding claim, wherein the subject is a pediatric, a teenager or an adult human.

35. The method of any preceding claim, wherein the test biological sample or the control biological sample each independently comprises a blood serum sample, a blood plasma sample, or a sample suitable for transcriptomic analysis.

36. The method of claim 35, wherein the test biological sample and the control biological sample are the same type of biological samples, or wherein the test biological sample and the control biological sample are different types of biological samples.

37. The method of any one of claims 19-22, wherein the therapeutically effective amount of the Sigma-1 receptor agonist ranges from about 0.5 to about 100 mg.

38. The method of any one of claims 19-22, wherein the therapeutically effective amount of the Sigma-1 receptor agonist ranges from about 5 mg to about 60 mg when administered orally.

39. The method of any one of claims 19-22, wherein the therapeutically effective amount of the Sigma-1 receptor agonist ranges from about 1 to about 20 mg, or from about 5 mg to about 30 mg, when administered intravenously.151108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center 40. The method of any one of claims 19-22, wherein the therapeutically effective amount of the Sigma-1 receptor agonist ranges from about 40 mg / day to about 60 mg / day.

41. The method of any one of claims 19-22, wherein the therapeutically effective amount provides a blood concentration of the Sigma-1 receptor agonist of about 4 ng / mL.

42. The method of claim 12, wherein the alternative to Sigma-1 receptor agonist is galantamine, memantine, rivastigmine, or combinations thereof.

43. The method of claim 26, wherein the condition is early onset Alzheimer’s disease or dementia.

44. A method of treating, preventing or delaying onset or occurrence of a neurodegenerative or neurodevelopmental disorder in a subject in need thereof, comprising administering a therapeutically effective amount of a Sigma-1 receptor agonist to the subject, wherein the subject carries one or more of the APOE4 gene, the COL24A1 gene, the ADRA1 D gene, and the ANXA2 gene; and wherein the neurodegenerative or neurodevelopmental disorder is pathologically associated with the APOE4 gene.

45. The method of claim 44, wherein the therapeutically effective amount is the amount effective in regulating lipid metabolism associated with the neurodegenerative or neurodevelopmental disorder.

46. The method of claim 44 or claim 45, wherein the subject suffers from or is suspected of suffering from the neurodegenerative or a neurodevelopmental disorder.

47. The method of any one of claims 44-46, wherein the neurodegenerative or neurodevelopmental disorder is selected from Alzheimer’s disease, Parkinson’s disease, dementia, Huntington’s disease, Amyotrophic lateral sclerosis, Prion152108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center disease, Rett Syndrome, cerebral palsy, Angelman syndrome, Williams syndrome, pervasive developmental disorder not otherwise specified (PDD-NOS), childhood disintegrative disorder, Smith-Magenis syndrome or multiple sclerosis, or any combination thereof.

48. The method of any one of claims 44-47, wherein the neurodegenerative or neurodevelopmental comprises early onset Alzheimer Disease or early stage Alzheimer Disease.

49. The method of any one of claims 44-48, wherein the Sigma-1 receptor agonist is selected from:tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine in amorphous form;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine in crystal Form I;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride in crystal Form I;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride in crystal Form II;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride in crystal Form III;(+) tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride in amorphous form;(-) tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride in amorphous form;(+) tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride in a crystal form;153108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center(-) tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride in a crystal form;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrogen fumarate in amorphous form;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrogen fumarate in crystal Form I;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrogen fumarate in crystal Form II;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrogen fumarate in crystal Form III;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrogen fumarate in crystal Form IV;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrogen fumarate in crystal Form V;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine mesylate in crystal Form I;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine sulfate in crystal Form I;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine sulfate in crystal Form II;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine oxalate in crystal Form I;154108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Centertetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine oxalate in crystal Form II;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine dihydrogen phosphate in crystal Form I;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine edisylate in crystal Form I;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine benzoate in crystal Form I;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrobromide in crystal Form A;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrobromide in crystal Form B;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine maleate in crystal Form S5;tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine maleate in crystal Form S6;co-crystal Form CSII of tetrahydro-N, N-dimethyl-2,2-diphenyl-3- furanmethanamine hydrochloride with tartaric acid;co-crystal Form CSIII of tetrahydro-N, N-dimethyl-2,2-diphenyl-3- furanmethanamine hydrochloride with citric acid;co-crystal Form CSIV of tetrahydro-N, N-dimethyl-2,2-diphenyl-3- furanmethanamine hydrochloride with malic acid;155108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Centera co-crystal of tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride with zinc chloride;a co-crystal of (-) tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride with zinc chloride;a co-crystal of (+) tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride with zinc chloride; andany combination thereof.

50. The method of any oy one of claims 44-48, wherein the Sigma-1 receptor agonist is selected from:tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride (A2- 73);tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine (A2-73 freebase);tetrahydro-N, N-dimethyl-5,5-diphenyl-3-furanmethanamine hydrochloride (A1- 41);tetrahydro-N, N-dimethyl-5,5-diphenyl-3-furanmethanamine (A1-41 freebase); 1-(2,2-diphenyltetrahydrofuran-3-yl)-N-methylmethanamine hydrochloride (A19-144);1-(2,2-diphenyltetrahydrofuran-3-yl)-N-methylmethanamine (A19-144 freebase);(1-(2,8-dimethyl-1-thia-3,8-diazaspiro(4.5)dec-3-yl)-3-(1H-indol-3-yl)propan-1-one) (A3-71);an enantiomer thereof, a crystal form thereof, a co-crystal form thereof, a pharmaceutically acceptable salt thereof, a crystal or co-crystal of the pharmaceutically acceptable salt thereof, and any combination thereof.156108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center 51. The method of any one of claims 44-48, wherein the Sigma-1 receptor agonist is tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine in Crystal Form I.

52. The method of any one of claims 44-48, wherein the Sigma-1 receptor agonist is tetrahydro-N, N-dimethyl-2,2-diphenyl-3-furanmethanamine hydrochloride in Crystal Form I.

53. The method of any one of claims 44-52, wherein the administering is selected from an oral administration, a subcutaneous administration, an intravenous administration, an intraocular administration, an intradermal administration, an intramuscular administration, an intraperitoneal administration, an intratracheal administration, an inhalation administration, an intranasal administration, a sublingual administration, a buccal administration, a rectal administration, a vaginal administration, and a topical administration.

54. The method of any one of claims 44-53, wherein the therapeutically effective amount is about 0.5 mg to about 100 mg per day.

55. The method of any one of claims 44-54, wherein the therapeutically effective amount is from about 1 mg to about 60 mg per day when administered orally.

56. The method of any one of claims 44-55, wherein the therapeutically effective amount is from about 0.5 mg to about 30 mg per day when administered intravenously.

57. The method of any one of claims 44-55, wherein the therapeutically effective amount is selected from about 5 mg, about 10 mg, about 20 mg, about 30 mg, about 40 mg, about 50 mg, and about 60 mg per day.

58. The method of any one of claims 44-57, wherein the administering lasts for at least about 30 weeks.157108513585 1PATENT Attorney Docket No.: 085102-875342Via Patent Center 59. The method of any one of claims 44-57, wherein the administering lasts for at least about 48 weeks.

60. The method of any one of claims 44-57, wherein the administering is daily administration.

61. The method of any one of claims 44-57, wherein the administering is intermittent administration with alternating dosing and resting periods.

62. The method of any one of claims 44-61, wherein the Sigma-1 receptor agonist is selected from the group consisting of amorphous form of A2-73 free base, a crystal form of A2-73 freebase, A2-73 amorphous form, a A2-73 crystal form I, or a A2-73 enantiomer.

63. The method of any one of claims 44-62, wherein the Sigma-1 receptor agonist is A2-73 Crystal Form I, (-) A2-73 enantiomer, or any combination thereof.

64. The method of any one of claims 44-63, wherein the Sigma-1 receptor agonist is A2-73 in an oral dosage form, and wherein the A2-73 is administered once daily for a period of at least 48 weeks.

65. The method of any one of claims 44-64, wherein the subject is an adult human less than 65 years old.

66. The method of any one of claims 44-65 wherein the neurodegenerative or neurodevelopmental disorder is early onset Alzheimer’s Disease.158108513585 1

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