Treatment of neurological diseases

Abstract

The present invention relates to (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol for the treatment of diseases mediated by protein expression and gene regulation, and to the use of personalized medicine approaches.

Description

Technical Field

[0001] Related Art This application claims the priority of U.S. Provisional Patent Application No. 63 / 529,307, filed Jul. 27, 2023, the entire content of which is incorporated herein by reference.

[0002] Field The present invention is directed to (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol for the treatment of diseases mediated by protein expression and gene regulation, as well as the use of personalized medical approaches.

Background Art

[0003] / / No content here, so it remains as is / / No content here, so it remains as is The therapeutic targeting of oxidative stress in amyotrophic lateral sclerosis (ALS) has not made much progress to clinical benefit for patients to date. This may be rationalized by the current use of antioxidants that target downstream biology. An alternative approach is to target the entire biological pathway by engaging the master regulator, the transcription factor nuclear factor erythroid 2-related factor 2 (NRF2), and thus involves the activation of several neuroprotective pathways (Nguyen et al., Annu. Rev. Pharmacol. Toxicol., 2003. 43:233 - 60). Under normal conditions, NRF2 activation is maintained at low levels through negative regulation by the Kelch-like ECH-associated protein 1 (Keap1) complex. Upon exposure to oxidative stress, cysteine residues on Keap1 are oxidized, dissociating the interaction between NRF2 and Keap1 (Wakabayashi et al., Proc Natl Acad Sci U S A., 2004 17;101(7):2040 - 5). This prevents the ubiquitination of NRF2, allows its nuclear translocation, and drives the expression of detoxification and antioxidant enzymes through its interaction with antioxidant response elements (ARE) on multiple cytoprotective genes.

[0004] NRF2 expression activates a "programmed cell survival" response, upregulating cytoprotective and antioxidant genes, as well as genes related to glutathione (GSH) synthesis, nicotinamide adenine dinucleotide phosphate (NADPH) production, and lipid and glucose / glycogen metabolism, all of which are neuroprotective. Downregulation of NRF2 has also been reported in both mouse models and human cases of sporadic ALS (sALS). Previous studies have demonstrated the beneficial effects of NRF2 activation on ALS models. In particular, astrocyte-specific expression of NRF2 delayed disease onset and extended survival in SOD1 transgenic mouse models. Theoretically, NRF2-activating compounds may protect against astrocyte toxicity to MN and delay disease progression in ALS patients.

[0005] Whole-genome expression profiling is a widely used tool for investigating the transcriptome. Changes in gene expression in ALS have been explored using microarrays, libraries for simultaneously detecting the expression of thousands of genes, or mRNA sequencing (Cooper-Knock et al. Neurology., 2012, 8(9), 518-530). While transcriptome analysis provides a good indicator of differential gene expression (DGE), the significance of the data is limited because mRNA levels do not necessarily correlate with the levels of translated proteins. These fluctuations can be attributed to proteolysis, oxidative stress, or regulation of protein synthesis. Therefore, translatome profiling, i.e., sequencing of mRNA recruited to ribosomes for protein synthesis, would likely better reflect changes in protein expression and the direction of disease processes in ALS-induced astrocytes (iAstrocytes).

[0006] There are three main methodologies for translatome profiling: polysome profiling, ribosome profiling, and, more recently, ribosome affinity purification techniques. Early methods for studying the overall translatome were performed by comparing ribosome-bound mRNA with the total mRNA present in the sample. Polysome profiling is a classic mRNA extraction technique involving the separation of mRNA according to the number of bound ribosomes (polysomes) using a sucrose gradient. While polysome profiling was considered the "gold standard" for many years, the use of a sucrose gradient required specialized and expensive equipment, as well as an additional precipitation step, due to the presence of heparin, a potent RNAse inhibitor, in the sucrose solution. Polysome fractions can also be contaminated with other high molecular weight complexes, such as lipid rafts or pseudopolysomes.

[0007] Ribosome profiling is based on sequencing of ribosome protection fragments (RPFs) after RNase I treatment of cell lysates. This methodology operates on the premise that the average ribosome density per mRNA correlates with the level of protein synthesis. For translatome analysis, mRNA extraction and sequencing are performed in parallel to normalize RPFs against total mRNA. Because RNase I degrades only single-stranded RNA, leading to misinterpretation of data, this method has its own challenges, including sucrose gradients, labor-intensive methods, and potential contamination by false RPFs from structured double-stranded regions of RNA, similar to polysome profiling.

[0008] Ribosome affinity purification has become a common tool for monitoring gene expression in specific cell types, such as neurons, due to the difficulty in isolating these cells without contamination from surrounding cells or tissues. This method involves constructing genetically modified cells / organisms that express affinity-tagged ribosome subunits, which can be controlled by tissue-specific promoters. The tagged ribosomes are recovered by affinity selection, capturing pure ribosomes from the target cells. RNA is isolated from the captured ribosomes and measured via microarray / RNA sequencing. While this method offers the best high-throughput prospects compared to polysome and ribosome profiling, the technique remains limited by the lack of distinction between actively translating mRNA and untranslated mRNA, which may be either mRNA indirectly bound to the ribosome or mRNA bound to another untranslated mRNA.

[0009] A novel method of translatome profiling has been developed by the present inventors, which enables the isolation of RNA molecules that are likely to co-precipitate with ribosomes and therefore undergo translation into proteins.

[0010] Whole-genome expression profiling can be used for personalized medicine. In the concept of personalized medicine, the "responder" subgroup is selected based on specific criteria. Strict cutoffs are chosen to avoid noise from mild responders. On the other hand, "non-responders" are the group that does not meet certain criteria and are compared to the "responder" group. Biomarkers are used to distinguish "responders" from the "non-responder" group, and each drug can obtain a more favorable risk-response ratio, enabling clinicians to make better treatment choices for patients.

[0011] Personalized medicine has been widely applied to specific types of cancer, and it is beginning to be applied to the treatment of patients with Alzheimer's disease and multiple sclerosis (MS) (Krzyszczyk et al., Technology, 2018, 6(3-4), 79-100; Hampel et al., The Journal of Prevention of Alzheimer's Disease, 2016, 3(4), 243-259; Gafson et al., Multiple Sclerosis (Houndmills, Basingstoke, England), England, 2016, 23(3), 362-369). Over the past decade, evidence has accumulated that no two patients' cancers are identical. This has resulted in different patient-to-patient responses to traditional cancer therapies such as radiation and chemotherapy. The application of personalized medicine approaches has led to the development of specialized treatments for each subtype of cancer based on patient genetic data, including transcriptomics, metabolomics, and proteomics. One example of this is the discovery of mutations in anaplastic lymphoma kinase (ALK) that drive tumorigenesis in approximately 5% of non-small cell lung cancers (Soda et al., Nature. 2017, 448(7153), 561-566). Following the identification of these mutations, ALK blockers (e.g., crizotinib and certinib) were developed and specifically administered to patients who tested positive for ALK mutations.

[0012] Multiple sclerosis (MS) is a syndrome characterized by significant variability in clinical phenotype, disease onset, and treatment response. With the increasing number of available treatments for MS patients, all of which involve different mechanisms of action, ranges of efficacy, and relative risks, a personalized, tailored approach must be adapted to make the best decisions for each individual patient. In MS, the clinical picture, along with clinical laboratory tests, imaging, and CSF testing, is used collectively to define each patient's syndrome by rejecting possible alternative disorders and recognizing distinct sub-syndromes of primary progressive MS. Neuromyelitis optica spectrum disorder (NMOSD) can be identified by serum antibodies against aquaporin 4 (AQP4-IgG). It is important to differentiate this disorder from MS, as patients with NMOSD may not respond to or may experience exacerbation with interferon (IFN) treatment.

[0013] In contrast, personalized medicine approaches have not been applied to many other neurodegenerative diseases, including ALS. Similar to MS, ALS is a heterogeneous neurodegenerative disease urgently requiring new treatments. Understanding the heterogeneity of ALS involves the deconstruction of biologically important pathways leading to the disease and how patient-specific factors influence these pathways. Here, we identify disease-specific pathways or transcriptional signatures that may aid in selecting the right drugs to administer to patients and in identifying novel compounds that target the disease in specific patients. We also identify a panel of biomarkers for drug response by investigating how the significance of differentially expressed genes (DEGs) in patient iAstrocytes changed after drug treatment, i.e., whether transcripts that were significantly dysregulated at baseline were no longer significantly dysregulated after treatment. To investigate changes in individual genes, rigorous statistical analysis was applied to reduce the probability of accepting false-positive results. The false detection rate (FDR) was applied to generate corrected p-values ​​(p-adj). Specifically, p-adj<0.05 means that the inventors accepted the probability that 5% of the already selected DEGs (p<0.05) were false positives. [Overview of the project]

[0014] (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol, the enantiomer of the currently approved (6aR)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol, is a weak dopamine antagonist and does not exhibit dopamine agonism-related side effects after administration. Also known as S-(+)-10,11-dihydroxyaporfin, (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol is described by the following chemical structure: [ka]

[0015] This disclosure provides that (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol can significantly increase p62 protein, increase LC3 protein, decrease mitochondrial morphogenetic factors, alter biological pathways, and alter gene expression levels.

[0016] (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol may be used in methods to increase p62 protein, increase LC3 protein, decrease mitochondrial morphogenetic factors, alter biological pathways, and alter levels of gene expression in cells. (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol may be further used in methods to treat diseases mediated by p62 protein, LC3 protein, and mitochondrial morphogenetic factors.

[0017] In one embodiment, disclosed herein is a method for increasing the level of p62 protein in cells, comprising the step of contacting cells with an effective amount of (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol.

[0018] In one embodiment, disclosed herein is a method for increasing the level of LC3 protein in cells, comprising the step of contacting cells with an effective amount of (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol.

[0019] In one embodiment, the LC3 protein is the LC3-I protein. In another embodiment, the LC3 protein is the LC3-II protein.

[0020] In one embodiment, disclosed herein is a method for reducing the levels of mitochondrial morphological factors in cells, comprising the step of contacting cells with an effective amount of (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol.

[0021] In one embodiment, disclosed herein is a method for altering a biological pathway in a cell, comprising the step of contacting the cell with an effective amount of (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol.

[0022] In one embodiment, the biological pathway may represent a cellular response to an organic cyclic compound, a MAPK cascade, epithelial fold morphogenesis, cell adhesion, angiogenesis, an inflammatory response, an immune response, a redox process, a xenobiotic metabolic process, cholinergic synaptic transmission, or collagen fiber organization.

[0023] In one embodiment, a method for altering the level of gene expression in cells is disclosed herein, comprising the step of contacting cells with an effective amount of (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol.

[0024] In one embodiment, the genes are HS6ST2-AS1, CYP1B1, MEGF6, AHRR, CILP, TNS4, CACNA1S, IFNLR1, KSR2, OLFML2B, BCL11B, FGF18, GALNT5, NKAIN1, DAPP1, LINC00599, GPAT3, IL1B, PRAL, FXYD3, CYP26B1, CYP1B1-AS1, CH25H, COLEC12, TIPARP, KIAA1549L, IFI30, NPTX1, ZMIZ1-AS1, ANPEP, ARHGEF16, WFDC11, KIAA1549, SUSD3, H19, GFRA1, NRG1, NQO1, TCF7, DTX1, PRXL2A, COLGALT2, LUM, HSPB6, SLC16A10, COL24A1, KCND3, SLC1A2, WNT11, CLDN7, RNF128, PTGS1, ZNF385D, ADGRE2, LINC00639, ADGRD1, PRELP, TGFBI, FAM47E-STBD1, TSPAN15, SLC2A1, IER3, AJUBA, NTN4, CEMIP, CX3CL1, SLC16A3, STC2, THBD, IGFBP2, TNFRSF1B, ANXA3, NRP1, CD24-eight, CDC42EP2, IGSF3, ETNPPL, APCDD1, GNAZ, S1PR2, STK26, RDH10, RRAD, TNFAIP2, IL15RA, ITGB4, CD14, CAMK2B, SYNJ2, PIR, FAM156A, TPRA1, F2R, RGL1, GATAD2B, SLC6A6, NEDD4L, MN1, ATF3, ADCY9, SASH1, SLC22A23, SLC7A5, ANXA11, ZNF70, CD58, IL6R, ATXN1, FHL2, GABRA2, CSGALNACT1, S100A4, BIN1, PAPSS2, LMO4, RAB31, EPM2A, TP53INP1, SESN3, LGI2, C14orf132, ENHO, GCSHP5, LRATD1, LGALS7B, MARCHF3, CRYAB, ID1, PLPP4, RPSAP58, EEF1A1P5, TPPP3, MBNL1-AS1, CCDC171, TMEM189-UBE2V1, AFF2, SCIN, ID3, RUBCNL, EFEMP1, MYRIP, MEST, MDFI, LY6D, RHBDL3, SAMD11, OR2S1P,It is selected from one or more of HSPE1-MOB4, LGALS7, HPGD, SULT1E1, LINC00598, or ONECUT2.

[0025] In another embodiment, the gene is selected from one or more of ICA1, MRPS9, NRIP3, NECTIN3, GATD3A, COL5A3, EPHA3, MFF_DT, IGFBP5, ROCK1P1, LYPD6, DKK2, CAP2, or GRM4.

[0026] As used herein, the term "effective amount" means an amount that produces a desired effect or result, for example, an amount that results in increasing the level of p62, increasing the level of LC3, decreasing the mitochondrial morphological factor, changing a biological pathway, and / or changing the level of gene expression.

[0027] In one embodiment, the method can be an in vitro method.

[0028] In another aspect, disclosed herein is a method of increasing the level of p62, increasing the level of LC3, decreasing the mitochondrial morphological factor, changing a biological pathway, and / or changing the level of gene expression in a cell, the method comprising contacting the cell with an effective amount of (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol.

[0029] In one embodiment, the method can be an in vitro method.

[0030] In one embodiment, the cells in the above embodiment, or in one of the other embodiments herein, are derived from a cell type or tissue selected from one or more of the following: adrenal gland, bone marrow, brain, breast, bronchi, caudate nucleus, cerebellum, cerebral cortex, cervix, uterus, colon, endometrium, epididymis, esophagus, fallopian tube, gallbladder, myocardium, hippocampus, kidney, liver, lung, lymph node, nasopharynx, oral mucosa, ovary, pancreas, parathyroid gland, peripheral blood mononuclear cells (PBMCs), placenta, prostate, rectum, salivary gland, seminal vesicle, skeletal muscle, skin, small intestine (including duodenum, jejunum, and ileum), smooth muscle, spleen, stomach, testis, thyroid gland, tonsil, bladder, and vagina. In further embodiments, brain cells are derived from brain tissue selected from the cerebrum (including the cerebral cortex, basal ganglia (often called the striatum), and olfactory bulb), cerebellum (including the dentate nucleus, intermediate nucleus, fastigial nucleus, and vestibular nuclei), diencephalon (including the thalamus, hypothalamus, etc., and the posterior part of the pituitary gland), and brainstem (including the pons, substantia nigra, and medulla oblongata). In further embodiments, brain cells are selected from neurons or glial cells (e.g., astrocytes, oligodendrocytes, or microglia). In further embodiments, neurons are sensory neurons, motor neurons, interneurons, or cerebral neurons.

[0031] In one embodiment, the cells are animal cells, for example, mammalian cells. In a further embodiment, the cells are human cells or non-human cells. In a further embodiment, the cells are in vitro, in vivo, or ex vivo.

[0032] In another embodiment, the cells are disease cells. In another embodiment, the cells are disease cells derived from a patient suffering from a disease or disorder as defined below.

[0033] In another embodiment, disclosed herein is a method for treating an animal having a disease or disorder which would benefit from increasing the level of p62, increasing the level of LC3, decreasing mitochondrial morphological factors, altering biological pathways, or altering the level of gene expression, the method comprising administering to the animal a therapeutically effective amount of a pharmaceutical composition comprising (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol.

[0034] In another embodiment, disclosed herein is (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol for use in the treatment of disease or disorder by increasing p62 levels, increasing LC3 levels, decreasing mitochondrial morphogenetic factors, altering biological pathways, or altering gene expression levels.

[0035] In one embodiment, (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol may be used for the treatment of animals having a disease or disorder characterized by increasing p62 levels, increasing LC3 levels, decreasing mitochondrial morphogenetic factors, altering biological pathways, or altering gene levels.

[0036] In one embodiment, (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol may be included in a pharmaceutical composition.

[0037] In one embodiment, a pharmaceutical composition containing (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol, or (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol, may be used for administration to animals in an effective amount.

[0038] In one embodiment, the animal is a mammal. In another embodiment, the mammal is a human or a non-human mammal. In a further embodiment, the mammal is a human.

[0039] In another embodiment, the disease or disorder is caused by dysregulation of p62, LC3, mitochondrial morphological factors, biological pathways, and / or genes.

[0040] In another embodiment, the disease is a neurodegenerative disease.

[0041] In another embodiment, the disease is age-related macular degeneration, Alzheimer's disease (AD), amyotrophic lateral sclerosis (ALS) / motor neuron disease (MND), atherosclerosis, autism spectrum disorder (ASD), benign focal muscular atrophy, cerebral infarction, Creutzfeldt-Jakob disease, Crohn's disease, Duchenne muscular dystrophy, Friedreich's ataxia, frontotemporal dementia (FTD), glaucoma, hereditary spastic paraplegia (HSP), Huntington's disease (HD), inclusion body myositis (IBM), inflammatory bowel disease, ischemia, Kugelberg-Welander syndrome, Lewy body dementia (LBD), Lou Gehrig's disease, multiple sclerosis (MS), myocardial infarction, necrotizing enterocolitis, neurofibromatosis type 1, Paget's disease of bone (PDB), and Parkinson's disease. (PD), primary lateral sclerosis (PLS), progressive bulbar palsy (PBP), progressive muscular atrophy (PMA), pseudobulbar palsy, spinal muscular atrophy (SMA), ulcerative colitis, balocin-containing protein (VCP) related disorder, or one or more of the following: Werdnig-Hoffmann disease, transient ischemic attack, ischemia, cerebral hemorrhage, senile cataract, retinal ischemia, retinal vasculitis, Brown-Bearet-van Lehr syndrome, Eels disease, meningitis and encephalitis, post-traumatic stress disorder, Charcot-Marie-Tooth disease, macular degeneration, X-linked spinal muscular atrophy (Kennedy disease), presenile dementia, depressive disorder, temporal lobe epilepsy, hereditary Leber's optic nerve atrophy, cerebrovascular disease, subarachnoid hemorrhage, and schizophrenia.

[0042] In one embodiment, the disease is amyotrophic lateral sclerosis (ALS).

[0043] In one embodiment, the disease is ALS caused by a mutation. In one embodiment, the disease is ALS caused by a mutation selected from the C9orf72 mutation, the SOD1 mutation, or another rarer mutation that causes or makes a subject with the mutation susceptible to ALS.

[0044] In another embodiment, disclosed herein is an in vitro method for screening therapeutic candidate(s) for their ability to treat animals, the method being: (1) Exposing animals, for example, animal cells, to a candidate drug; (2) Compare the levels of gene expression in cells before and after treatment after a certain period of time; (3) Based on the comparison results, this includes determining whether the animals can benefit from treatment with the candidate drug(s).

[0045] In some embodiments, the methods described herein involve comparing the levels of gene expression in cells before and after treatment, where the therapeutic agent candidate is (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol or a salt thereof. In some embodiments, the comparison involves comparing or determining the expression of p62 protein, LC3 protein, or mitochondrial morphological factors, or significant changes in their expression. In some embodiments, the comparison involves comparing or determining the expression of biological pathways, or significant changes in biological pathways, or changes in the level of gene expression.

[0046] In one embodiment, the method may be an in vitro method.

[0047] In one embodiment, the period of exposure to cells is 1 to 30 days, preferably 1 to 15 days, and preferably 1 to 5 days.

[0048] In one embodiment, the candidate therapeutic agent is (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol.

[0049] In one embodiment, the animal is a mammal. In another embodiment, the mammal is a human or a non-human mammal. In a further embodiment, the mammal is a human.

[0050] In one embodiment, the cells in one of the above embodiments or in other embodiments herein are derived from a cell type or tissue selected from one or more of the following: adrenal gland, bone marrow, brain, breast, bronchi, caudate nucleus, cerebellum, cerebral cortex, cervix, uterus, colon, endometrium, epididymis, esophagus, fallopian tube, gallbladder, myocardium, hippocampus, kidney, liver, lung, lymph node, nasopharynx, oral mucosa, ovary, pancreas, parathyroid gland, peripheral blood mononuclear cells (PBMCs), placenta, prostate, rectum, salivary gland, seminal vesicle, skeletal muscle, skin, small intestine (including duodenum, jejunum, and ileum), smooth muscle, spleen, stomach, testis, thyroid gland, tonsil, bladder, and vagina. In further embodiments, the brain cells are derived from brain tissue selected from the cerebrum (including the cerebral cortex, basal ganglia (often called the striatum), and olfactory bulb), cerebellum (including the dentate nucleus, intermediate nucleus, fastigial nucleus, and vestibular nuclei), diencephalon (including the thalamus, hypothalamus, etc., and the posterior part of the pituitary gland), and brainstem (including the pons, substantia nigra, and medulla oblongata). In further embodiments, the brain cells are selected from neurons or glial cells (e.g., astrocytes, oligodendrocytes, or microglia). In further embodiments, the cells are peripheral blood mononuclear cells (PBMCs).

[0051] In one embodiment, the cells are animal cells, for example, mammalian cells. In a further embodiment, the cells are human cells or non-human cells. In a further embodiment, the cells are in vitro, in vivo, or ex vivo.

[0052] In another embodiment, the cells are disease cells. In another embodiment, the cells are disease cells derived from a patient suffering from a disease or disorder as defined below.

[0053] In one embodiment, the genes are HS6ST2-AS1, CYP1B1, MEGF6, AHRR, CILP, TNS4, CACNA1S, IFNLR1, KSR2, OLFML2B, BCL11B, FGF18, GALNT5, NKAIN1, DAPP1, LINC00599, GPAT3, IL1B, PRAL, FXYD3, CYP26B1, CYP1B1-AS1, CH25H, COLEC12, TIPARP, KIAA1549L, IFI30, NPTX1, ZMIZ1-AS1, ANPEP, ARHGEF16, WFDC11, KIAA1549, SUSD3, H19, GFRA1, NRG1, NQO1, TCF7, DTX1, PRXL2A, COLGALT2, LUM, HSPB6, SLC16A10, COL24A1, KCND3, SLC1A2, WNT11, CLDN7, RNF128, PTGS1, ZNF385D, ADGRE2, LINC00639, ADGRD1, PRELP, TGFBI, FAM47E-STBD1, TSPAN15, SLC2A1, IER3, AJUBA, NTN4, CEMIP, CX3CL1, SLC16A3, STC2, THBD, IGFBP2, TNFRSF1B, ANXA3, NRP1, CD248, CDC42EP2, IGSF3, ETNPPL, APCDD1, GNAZ, S1PR2, STK26, RDH10, RRAD, TNFAIP2, IL15RA, ITGB4, CD14, CAMK2B, SYNJ2, PIR, FAM156A, TPRA1, F2R, RGL1, GATAD2B, SLC6A6, NEDD4L, MN1, ATF3, ADCY9, SASH1, SLC22A23, SLC7A5, ANXA11, ZNF70, CD58, IL6R, ATXN1, FHL2, GABRA2, CSGALNACT1, S100A4, BIN1, PAPSS2, LMO4, RAB31, EPM2A, TP53INP1, SESN3, LGI2, C14orf132, ENHO, GCSHP5, LRATD1, LGALS7B, MARCHF3, CRYAB, ID1, PLPP4, RPSAP58, EEF1A1P5, TPPP3, MBNL1-AS1, CCDC171, TMEM189-UBE2V1, AFF2, SCIN, ID3, RUBCNL, EFEMP1, MYRIP, MEST, MDFI, LY6D, RHBDL3, SAMD11, OR2S1P,Select one or more from HSPE1-MOB4, LGALS7, HPGD, SULT1E1, LINC00598, and ONECUT2.

[0054] In another embodiment, the gene is selected from one or more of ICA1, MRPS9, NRIP3, NECTIN3, GATD3A, COL5A3, EPHA3, MFF_DT, IGFBP5, ROCK1P1, LYPD6, DKK2, CAP2, and GRM4.

[0055] The above and other features and advantages of the present invention will become more apparent from the following detailed description, which proceeds with reference to the accompanying drawings. Such description is intended to be illustrative of the invention and not limiting. Obvious variations of thiazolidinedion disclosed herein, including those described by the drawings and embodiments, will be readily apparent to those skilled in the art having this disclosure, and such variations will be considered part of the present invention.

[0056] In all figures disclosed herein, dexamethasone, rosiglitazone, pioglitazone, and thiazolidinedione are abbreviated as Dex, Rosi, Pio, and TZD, respectively. [Brief explanation of the drawing]

[0057] [Figure 1] This figure illustrates the direct conversion of ALS patient fibroblasts into iNPCs. Fibroblasts were transduced using a retroviral vector containing reprogramming factors Oct4, Sox2, Klf4, and c-Myc, and supplemented with NPC medium and growth factors. The cells were grown until the 18-day mark when iNPCs were obtained. [Figure 2A-2B]Quantification of mouse motor neuron rescue in co-culture with induced astrocytes derived from healthy controls and ALS patients (CTR: pooled data from 3 healthy controls; ALS patients with C9orf72 mutations: C9orf72_183, C9orf72_78, and C9orf72_201; ALS patients with SOD1 mutations: SOD1_210, SOD1_102, and SOD1_100; sporadic ALS patients: sALS_17, sALS_12, and sALS_009). Changes in motor neuron survival using 5 μM or 10 μM androgravold (Figure 2A) and (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol (labeled as Drug) (Figure 2B) were compared with vehicle (DMSO). [Figure 3A-3B] Quantification of mouse motor neuron rescue in co-culture with induced astrocytes derived from healthy controls and ALS patients (CTR: pooled data from 3 healthy controls; ALS patients with C9orf72 mutations: C9orf72_183, C9orf72_78, and C9orf72_201; ALS patients with SOD1 mutations: SOD1_210, SOD1_102, and SOD1_100; sporadic ALS patients: sALS_17, sALS_12, and sALS_009). Changes in motor neuron survival using 5 μM or 10 μM monomethyl fumarate (Figure 3A) and riluzole (Figure 3B) were compared with vehicle (DMSO). [Figure 4]Quantification of mouse Hb9GFP+ motor neuron rescue in co-culture with induced astrocytes by increased percentage of motor neuron survival 3 days after administration of 10 μM riluzole, andrografolide, and (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol (labeled as Drug) compared to vehicle (DMSO). Human iAstrocytes were derived from the same diverse ALS patients: healthy controls and three different sporadic ALS patients (sALS, n=3); three different ALS patients with SOD1 mutations (SOD1, n=3); and three different ALS patients with C9orf72 mutations (C9orf, n=3). *p<0.05;**p<0.01;***p<0.001;****p<0.0001. [Figure 5] Representative images showing the expression of p62(568) and cytoplasmic marker CD44(488) and nuclei by Hoechst staining in control and patient cells before and after treatment with the NRF2 activating compound (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol and androgravolide. Scale bar 10 μm. [Figure 6A-6C] This figure shows the quantification of p62 expression in control and patient iAstrocytes before and after drug treatment. (Figure 6A) S[+]-apomorphine (two-way ANOVA, MC, n=3, technical replicates=2, row factor p<0.05, column factor p<0.0001), (Figure 6B) androgravoride (two-way ANOVA, MC, n=3, technical replicates=2, row factor p<0.0001, column factor p<0.001), and (Figure 6C) MMF (two-way ANOVA, MC, n=3, technical replicates=2, row factor p=0.1945, column factor p<0.0001). Individual significance levels are shown on the graph. The control bars consist of 3050 and 155 cells pooled together. [Figure 7A-7C]This figure shows the quantification of p62 expression in the perinuclear region of control and patient iAstrocytes before and after drug treatment. (Figure 7A) Number of perinuclear spots after treatment with (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol (two-way ANOVA, MC, n=3, technical replicates=2, row factor p<0.05, column factor p=0.1104), (Figure 7B) Number of perinuclear spots after treatment with andrographoride (two-way ANOVA, MC, n=3, technical replicates=2, row factor p<0.0001, column factor p=0.5064), and (Figure 7C) Number of perinuclear spots after treatment with MMF (two-way ANOVA, MC, n=3, technical replicates=2, row factor p=0.2456, column factor p<0.001). The control bars consist of 3050 and 155, pooled together. [Figure 8A-8B] This figure shows the quantification of p62 expression in the cytoplasm of control and patient iAstrocytes before and after riluzole treatment. (Figure 8A) Percentage of p62-positive cells (two-way ANOVA, MC, n=3, technical replicates=2, row factor p=0.8922, column factor p<0.001). (Figure 8B) Number of perinuclear spots (two-way ANOVA, MC, n=3, technical replicates=2, row factor p=0.2791, column factor p<0.01). The control bars consist of 3050 and 155 cells pooled together. [Figures 9A-9F]This figure shows the quantification of autophagy marker p62 and LC3-I / LC3-II protein expression in control and C9ORF72 patient iAstrocytes before and after drug treatment. (Figure 9A) Western blot of p62 expression in control and C9ORF72 iAstrocytes. (Figure 9B) Quantification of p62 protein expression (mean ± SD, n=2) after treatment with (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol and androgravold, and (Figure 9C) after treatment with androgravold. (Figure 9D) Western blot of LC3-I / LC3-II protein expression in control and C9ORF72 iAstrocytes. (Figure 9E) shows the quantification of the ratio between LC3-I / LC-3II protein expression after treatment with (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol and androgravolide, and (Figure 9F) after treatment with androgravolide (mean ± SD, n=2). The control bars consist of 3050 and 155, pooled together. [Figure 10] Visualization of mitochondria (TMRM) and nuclei (Hoechst) in control and patient iAstrocytes before and after treatment with androgravoride. Scale bar 10 μm. [Figure 11A-11C] Figure 11A shows the quantification of mitochondrial morphological factors in control and patient iAstrocytes before and after treatment with (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol (two-way ANOVA, MC, n=3, technical replicates=3, row factor p<0.001, column factor p<0.0001), (Figure 11B) MMF (two-way ANOVA, MC, n=3, technical replicates=3, row factor p<0.05, column factor p<0.001), and (Figure 11C) riluzole (two-way ANOVA, MC, n=3, technical replicates=3, row factor p<0.01, column factor p<0.001). The control bars consist of 3050, 155, and 209, pooled together. [Figures 12A-12B]This figure shows a visual representation of differentially expressed genes (DEGs) shared between controls and different patient subgroups after (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol treatment, generated from the comparisons performed in Table 1. (Figure 12A) Total number of genes shared between groups. (Figure 12B) Number of upregulated (upper number in each pair) and downregulated (lower number in each pair) transcripts shared between subgroups. S = (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol treatment; CRT = control; U = untreated. [Figure 13] Visual interpretation of DEGs shared between (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol response comparisons shown in Table 6. The number of upregulatory genes is shown in black, and downregulatory transcripts are shown in gray. Of the 14 genes (11 elevated, 3 decreased) that were specific to the baseline S Resp.U vs CTR U comparison, 10 of them were independently dysregulated in M102 responders at baseline and corrected after treatment with (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol. S = (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol. [Figure 14A-14C] These are normalized gene counts (TPM values) for patient responders and non-responders to (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol before and after drug treatment. (Figure 14A) Normalized gene counts for NECTIN3 (n=1), (Figure 14B) MFF-DT (n=1), and (Figure 14C) GRM4 (n=1). The control bars consist of 3050 and AG. [Figure 15]Summary of gene expression changes at each biomarker identified within the (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol treatment response group. Normalized gene counts were plotted to determine how transcript expression changed in each cell line after treatment with (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol. Each was scored based on the post-treatment response; change in the direction of the control (+1), no change (0), or change in the opposite direction of the control (-1), and the total for each cell line is presented at the bottom. [Modes for carrying out the invention]

[0058] The term "(6aR)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol" refers to R-(-)-10,11-dihydroxyaporphine and includes its prodrugs, salts, solvates, hydrates, and cocrystals.

[0059] The term "(6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol" refers to S-(+)-10,11-dihydroxyaporphine and includes its prodrugs, salts, solvates, hydrates, and cocrystals.

[0060] The term "6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol" is (6aR)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol, or (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,1 This refers to 1-diols, or racemates of (6aR)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol and (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol, and includes their prodrugs, salts, solvates, hydrates, and cocrystals.

[0061] As used herein, the terms “to treat,” “to treat,” or “treatment” mean to alleviate, reduce or eliminate one or more symptoms or characteristics of a disease, and may be therapeutic, palliative, preventive, or slow the progression of the disease.

[0062] The term “effective dose” means the amount that produces a desired effect or result, such as increasing p62 levels, increasing LC3 levels, decreasing mitochondrial morphogenetic factors, altering biological pathways, or altering gene levels. The term “therapeutic effective dose” means the amount of (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol that, alone or in combination with other active ingredients, elicits a desired biological or pharmacological response, for example, to prevent, alleviate, or improve the symptoms of a disease or disorder; to slow, halt, or reverse an underlying disease process or progression; to partially or completely restore cellular function; or to prolong the survival of the subject being treated.

[0063] The terms “patient” or “subject” include mammals, including non-human animals and especially humans. In one embodiment, the patient or subject is human. In another embodiment, the patient or subject is human male. In another embodiment, the patient or subject is human female.

[0064] The term "significant" or "significantly" is determined by a t-test at a significance level of 0.05.

[0065] This disclosure relates to a method of using (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol to increase p62 levels, increase LC3 levels, decrease mitochondrial morphogenetic factors, alter biological pathways, or alter gene levels in cells, tissues, or animals.

[0066] This disclosure further relates to a method of using (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol for the treatment, prevention, mitigation, or improvement of diseases mediated by p62 levels, LC3 levels, mitochondrial morphogenetic factors, biological pathways, or gene expression levels.

[0067] Accordingly, in one embodiment, the present disclosure provides a method for increasing the level of p62 protein in cells, the method comprising the step of contacting cells with an effective amount of (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol.

[0068] In one embodiment, the method may be an in vitro method.

[0069] In another embodiment, the p62 protein is a perinuclear p62 protein.

[0070] In a related embodiment, the present disclosure provides a method for increasing the level of LC3 protein in cells, the method comprising contacting cells with an effective amount of (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol.

[0071] In one embodiment, the method may be an in vitro method.

[0072] In another embodiment, the LC3 protein is either the LC3-I protein or the LC3-II protein.

[0073] In another embodiment, the present disclosure provides a method for reducing the levels of mitochondrial morphological factors in cells, the method comprising contacting cells with an effective amount of (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol.

[0074] In another embodiment, the present disclosure provides a method for altering a biological pathway in a cell, the method comprising contacting the cell with an effective amount of (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol.

[0075] In one embodiment, the biological pathway is a cellular response to an organic cyclic compound, a MAPK cascade, epithelial fold morphogenesis, cell adhesion, angiogenesis, inflammatory response, immune response, redox process, xenobiotic metabolic process, cholinergic synaptic transmission, or collagen fiber organization.

[0076] In another embodiment, the present disclosure provides a method for altering the level of gene expression in cells, the method comprising contacting cells with an effective amount of (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol.

[0077] In one embodiment, the genes are HS6ST2-AS1, CYP1B1, MEGF6, AHRR, CILP, TNS4, CACNA1S, IFNLR1, KSR2, OLFML2B, BCL11B, FGF18, GALNT5, NKAIN1, DAPP1, LINC00599, GPAT3, IL1B, PRAL, FXYD3, CYP26B1, CYP1B1-AS1, CH25H, COLEC12, TIPARP, KIAA1549L, IFI30, NPTX1, ZMIZ1-AS1, ANPEP, ARHGEF16, WFDC11, KIAA1549, SUSD3, H19, GFRA1, NRG1, NQO1, TCF7, DTX1, PRXL2A, COLGALT2, LUM, HSPB6, SLC16A10, COL24A1, KCND3, SLC1A2, WNT11, CLDN7, RNF128, PTGS1, ZNF385D, ADGRE2, LINC00639, ADGRD1, PRELP, TGFBI, FAM47E-STBD1, TSPAN15, SLC2A1, IER3, AJUBA, NTN4, CEMIP, CX3CL1, SLC16A3, STC2, THBD, IGFBP2, TNFRSF1B, ANXA3, NRP1, CD248, CDC42EP2, IGSF3, ETNPPL, APCDD1, GNAZ, S1PR, STK26, RDH10, RRAD, TNFAIP2, IL15RA, ITGB4, CD14, CAMK2B, SYNJ2, PIR, FAM156A, TPRA1, F2R, RGL1, GATAD2B, SLC6A6, NEDD4L, MN1, ATF3, ADCY9, SASH1, SLC22A23, SLC7A5, ANXA11, ZNF70, CD58, IL6R, ATXN1, FHL2, GABRA2, CSGALNACT1, S100A4, BIN1, PAPSS2, LMO4, RAB31, EPM2A, TP53INP1, SESN3, LGI2, C14orf132, ENHO, GCSHP5, LRATD1, LGALS7B, MARCHF3, CRYAB, ID1, PLPP4, RPSAP58, EEF1A1P5, TPPP3, MBNL1-AS1, CCDC171, TMEM189-UBE2V1, AFF2, SCIN, ID3, RUBCNL, EFEMP1, MYRIP, MEST, MDFI, LY6D, RHBDL3, SAMD11, OR2S1P,Select one or more from HSPE1-MOB4, LGALS7, HPGD, SULT1E1, LINC00598, and ONECUT2.

[0078] In another embodiment, the gene is selected from one or more of ICA1, MRPS9, NRIP3, NECTIN3, GATD3A, COL5A3, EPHA3, MFF_DT, IGFBP5, ROCK1P1, LYPD6, DKK2, CAP2, and GRM4.

[0079] In another embodiment, the Disclosure provides a method for (a) increasing the level of p62, (b) increasing the level of LC3, (c) decreasing mitochondrial morphogenetic factors, (d) altering a biological pathway, or (e) altering the level of a gene in a cell, the method comprising contacting a cell with an effective amount of (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol.

[0080] In one embodiment, the method may be an in vitro method.

[0081] In one embodiment, the cells in one of the above embodiments, or in other embodiments or embodiments of this specification, are derived from a cell type or tissue selected from one or more of the following: adrenal gland, bone marrow, brain, breast, bronchi, caudate nucleus, cerebellum, cerebral cortex, cervix, uterus, colon, endometrium, epididymis, esophagus, fallopian tube, gallbladder, myocardium, hippocampus, kidney, liver, lung, lymph node, nasopharynx, oral mucosa, ovary, pancreas, parathyroid gland, peripheral blood mononuclear cells (PBMCs), placenta, prostate, rectum, salivary gland, seminal vesicle, skeletal muscle, skin, small intestine (including duodenum, jejunum, and ileum), smooth muscle, spleen, stomach, testis, thyroid gland, tonsil, bladder, and vagina. In further embodiments, brain cells are derived from brain tissue selected from the cerebrum (including the cerebral cortex, basal ganglia (often called the striatum), and olfactory bulb), cerebellum (including the dentate nucleus, intermediate nucleus, fastigial nucleus, and vestibular nuclei), diencephalon (including the thalamus, hypothalamus, etc., and the posterior part of the pituitary gland), and brainstem (including the midbrain, pons, substantia nigra, and medulla oblongata). In further embodiments, brain cells are selected from neurons or glial cells (e.g., astrocytes, oligodendrocytes, or microglia). In further embodiments, neurons are sensory neurons, motor neurons, interneurons, or cerebral neurons. In some embodiments, brain cells are derived from brain tissue selected from the cerebrum, cerebellum, diencephalon, or brainstem. In some embodiments, brain cells are selected from neurons, astrocytes, oligodendrocytes, or microglia. In some embodiments, neurons are sensory neurons, motor neurons, interneurons, or cerebral neurons.

[0082] In some embodiments of the methods described herein, the cells are derived from a cell type or tissue selected from one or more of the following: adrenal gland, bone marrow, brain, breast, bronchi, caudate nucleus, cerebellum, cerebral cortex, cervix, uterus, colon, endometrium, epididymis, esophagus, fallopian tube, gallbladder, myocardium, hippocampus, kidney, liver, lung, lymph node, nasopharynx, oral mucosa, ovary, pancreas, parathyroid gland, peripheral blood mononuclear cells (PBMCs), placenta, prostate, rectum, salivary gland, seminal vesicle, skeletal muscle, skin, small intestine (including duodenum, jejunum, and ileum), smooth muscle, spleen, stomach, testis, thyroid gland, tonsil, bladder, or vagina.

[0083] In one embodiment, the cells are animal cells, for example, mammalian cells. In a further embodiment, the cells are human cells or non-human cells. In a further embodiment, the cells are human cells. In a further embodiment, the cells are in vitro, in vivo, or ex vivo.

[0084] In another embodiment, the cells are disease cells. In another embodiment, the cells are disease cells derived from a patient suffering from a disease or disorder disclosed herein.

[0085] In another embodiment, the Disclosure provides a method for treating an animal having a disease or disorder which would benefit from increasing the level of p62, increasing the level of LC3, decreasing mitochondrial morphogenetic factors, altering biological pathways, or altering gene levels, for example, wherein symptoms are prevented, reduced, or improved, or the disease process or progression is slowed, stopped, or reversed, the method comprising administering to the animal a therapeutically effective amount of a pharmaceutical composition comprising (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol.

[0086] In another embodiment, the Disclosure provides (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol for use in treating a disease or disorder by increasing the level of p62, increasing the level of LC3, decreasing mitochondrial morphogenetic factors, altering biological pathways, or altering the level of gene expression, for example, thereby preventing, reducing, or improving symptoms, or slowing, halting, or reversing a disease process or progression.

[0087] In one embodiment, (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol may be included in a pharmaceutical composition.

[0088] In one embodiment, a pharmaceutical composition containing (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol, or (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol, may be used for administration to animals in an effective amount.

[0089] In one embodiment, the animal is a mammal. In a further embodiment, the mammal is a human. In another embodiment, the mammal is a non-human mammal.

[0090] In another embodiment, the disease or disorder is caused at the level of p62, LC3, mitochondrial morphological factors, biological pathways, or genes.

[0091] In some embodiments, the disease or disorder is age-related tau astrocytosis (ARTA), Alexander disease, Alzheimer's disease, amyotrophic lateral sclerosis (ALS), critical disease myopathy (CIM), primary age-related tauopathy (PART), aortic medial amyloidosis, ApoAI amyloidosis, ApoAII amyloidosis, ApoAIV amyloidosis, argyrophilic granulosis, ataxia telangiectasia, atrial fibrillation, autosomal dominant hyper-IgE syndrome, atrial amyloidosis, Bloom syndrome, cardiovascular disease, coronary artery disease, myocardial infarction, stroke, Restenosis, arteriosclerosis, cataracts, cerebral amyloid angiopathy, Christianson syndrome, chronic traumatic encephalopathy, Cockayne syndrome, corneal lactoferrin amyloidosis, corticobasal degeneration, Crohn's disease, Cushing's disease, lichenoid amyloidosis, cystic fibrosis, dentatorubral-pallidoluysian atrophy (DRPLA), dialysis amyloidosis, diffuse neurofibrillary tangle disease with calcification, Down syndrome, endotoxin shock, Finnish familial amyloidosis, familial amyloid neuropathy, familial British dementia (FBD), familial Danish dementia Dementia (FDD), familial dementia, fibrinogen amyloidosis, fragile X syndrome, fragile X-associated tremor / ataxia syndrome (FXTAS), Friedreich's ataxia, frontotemporal degeneration, glaucoma, glycogen storage disease type IV (Andersen's disease), Guadeloupe-type parkinsonism, hereditary lattice keratitis dystrophy, Huntington's disease, inclusion body myositis / myopathy, inflammation, inflammatory bowel disease, ischemic state, ischemia / reperfusion injury, myocardial ischemia, stable angina, unstable angina, stroke, ischemic heart disease and cerebral ischemia, light chain or heavy chain amyloidosis, lysosomal storage disease, aspartylglucosa Minuria, Fabry disease, Batten disease, cystinosis, Faber disease, fucosidosis, galactosialidosis, Gaucher disease type 1, 2 or 3, GM1 gangliosidosis, Hunter disease, Haller-Scheye disease, Krabbe disease, α-mannosidosis, β-mannosidosis, Maloto-Lamy disease, metachromatic leukodystrophy, Morquio A syndrome, Morquio B syndrome, mucolipidosis type II, mucolipidosis type III, Niemann-Pick disease type A, B or C, Pompe disease, Sandhoff disease, Sanfilippo syndrome type A, B, C or D, Schindler's disease,Schindler-Kanzaki disease, sialidosis, Sly syndrome, Tay-Sachs disease, Wolmann disease, lysozyme amyloidosis, Mallory bodies, medullary thyroid carcinoma, mitochondrial myopathy, multiple sclerosis, multiple system atrophy, myotonic dystrophy, myotonic dystrophy, neurodegenerative diseases with cerebral iron deposition, neurofibromatosis, neuronal ceroid lipofuscinosis, odontogenic (Pinborg) tumor amyloid, Guam Parkinson's dementia complex, Parkinson's disease, peptic ulcer, Pick's disease, pituitary prolactinoma, post-encephalitis parkinsonism, prion disease (transmissible spongiform encephalopathy) (Creutzfeldt-Jakob disease (CJD), variant Creutzfeldt-Jakob disease, Gerstmann-Stroheimer's disease) One or more of the following conditions are selected: Isra Shainker syndrome, fatal familial insomnia (including Kuru), progressive supranuclear palsy, alveolar proteinosis, retinal ganglion cell degeneration in glaucoma, retinitis pigmentosa with rhodopsin mutations, seminal vesicle amyloidosis, senile systemic amyloidosis, serpinopathy, sickle cell disease, spinal and bulbar muscular atrophy (SBMA), spinocerebellar ataxia, spinocerebellar ataxia type 1, spinocerebellar ataxia type 2, spinocerebellar ataxia type 3 (Machado-Joseph disease), spinocerebellar ataxia type 6, spinocerebellar ataxia type 7, spinocerebellar ataxia type 8, spinocerebellar ataxia type 17), subacute sclerosing panencephalitis, tauopathy, type 2 diabetes mellitus, vascular dementia, or Werner syndrome.

[0092] In another embodiment, the disease is age-related macular degeneration, Alzheimer's disease (AD), amyotrophic lateral sclerosis (ALS), atherosclerosis, autism spectrum disorder (ASD), benign focal muscular atrophy, cerebral infarction, Creutzfeldt-Jakob disease, Crohn's disease, Duchenne palsy, Friedreich's ataxia, frontotemporal dementia (FTD), glaucoma, hereditary spastic paraplegia (HSP), Huntington's disease (HD), inclusion body myositis (IBM), inflammatory bowel disease, ischemia, Kugelberg-Wellander syndrome, Lewy body dementia (LBD), Lou Gehrig's disease, multiple sclerosis (MS), myocardial infarction, necrotizing enterocolitis, neurofibromatosis type 1, Paget's disease of bone (PDB), Parkinson's disease (PD), primary A selection of one or more of the following conditions: lateral sclerosis (PLS), progressive bulbar palsy (PBP), progressive muscular atrophy (PMA), pseudobulbar palsy, spinal muscular atrophy (SMA), ulcerative colitis, balocin-containing protein (VCP) related disorder, or Werdnig-Hoffmann disease, transient ischemic attack, ischemia, cerebral hemorrhage, senile cataract, retinal ischemia, retinal vasculitis, Brown-Bearet-van Lear syndrome, Eels disease, meningitis and encephalitis, post-traumatic stress disorder, Charcot-Marie-Tooth disease, macular degeneration, X-linked spinal muscular atrophy, presenile dementia, depressive disorder, temporal lobe epilepsy, hereditary Leber's optic nerve atrophy, cerebrovascular disease, subarachnoid hemorrhage, and schizophrenia.

[0093] In another embodiment, the disease is a neurological disorder.

[0094] In one embodiment, the disease is a neurodegenerative disease or disorder.

[0095] In one embodiment, the disease is ALS.

[0096] In one embodiment, the disease is ALS caused by a mutation. In one embodiment, the disease is ALS caused by a mutation selected from the C9orf72 mutation, the SOD1 mutation, or another mutation known to cause or predispose to ALS.

[0097] In some embodiments, methods are provided for treating animals having a disease or disorder which would benefit from altering a biological pathway in cells, the method comprising contacting cells with an effective amount of (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol, where the biological pathway is selected from one or more of the following: cellular response to an organic cyclic compound, MAPK cascade, epithelial fold morphogenesis, cell adhesion, angiogenesis, inflammatory response, immune response, redox process, xenobiotic metabolic process, cholinergic synaptic transmission, or collagen fiber organization.

[0098] In some embodiments, methods are provided for treating animals having a disease or disorder that would benefit from altering the level of a gene, the method comprising administering to the animal a therapeutically effective amount of a pharmaceutical composition comprising (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol, wherein the gene is HS6ST2-AS1, CYP1B1, MEGF6, AHRR, CILP, TNS4, CACNA1S, IFNLR1, KSR2, OLFML2B, BCL11B, FGF18, GAL NT5, NKAIN1, DAPP1, LINC00599, GPAT3, IL1B, PRAL, FXYD3, CYP26B1, CYP1B1-AS1, CH25H, COLEC12, TIPARP, KIAA1549L, IFI30, NPTX1, ZMIZ1-AS1, ANP EP, ARHGEF16, WFDC11, KIAA1549, SUSD3, H19, GFRA1, NRG1, NQO1, TCF7, DTX1, PRXL2A, COLGALT2, LUM, HSPB6, SLC16A10, COL24A1, KCND3, SLC1A2, WNT11 , CLDN7, RNF128, PTGS1, ZNF385D, ADGRE2, LINC00639, ADGRD1, PRELP, TGFBI, FAM47E-STBD1, TSPAN15, SLC2A1, IER3, AJUBA, NTN4, CEMIP, CX3CL1, SLC 16A3, STC2, THBD, IGFBP2, TNFRSF1B, ANXA3, NRP1, CD248, CDC42EP2, IGSF3, ETNPPL, APCDD1, GNAZ, S1PR2, STK26, RDH10, RRAD, TNFAIP2, IL15RA, ITGB4 , CD14, CAMK2B, SYNJ2, PIR, FAM156A, TPRA1, F2R, RGL1, GATAD2B, SLC6A6, NEDD4L, MN1, ATF3, ADCY9, SASH1, SLC22A23, SLC7A5, ANXA11, ZNF70, CD58, IL 6R, ATXN1, FHL2, GABRA2, CSGALNACT1, S100A4, BIN1, PAPSS2, LMO4, RAB31, EPM2A, TP53INP1, SESN3, LGI2, C14orf132, ENHO, GCSHP5, LRATD1, LGALS7B,Select one or more from the following: MARCHF3, CRYAB, ID1, PLPP4, RPSAP58, EEF1A1P5, TPPP3, MBNL1-AS1, CCDC171, TMEM189-UBE2V1, AFF2, SCIN, ID3, RUBCNL, EFEMP1, MYRIP, MEST, MDFI, LY6D, RHBDL3, SAMD11, OR2S1P, HSPE1-MOB4, LGALS7, HPGD, SULT1E1, LINC00598, or ONECUT2.

[0099] In some embodiments, methods are provided for treating animals having a disease or disorder that would benefit from altering the level of a gene, the method comprising administering to the animal a therapeutically effective amount of a pharmaceutical composition comprising (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol, wherein the gene is selected from one or more of ICA1, MRPS9, NRIP3, NECTIN3, GATD3A, COL5A3, EPHA3, MFF_DT, IGFBP5, ROCK1P1, LYPD6, DKK2, CAP2, or GRM4.

[0100] In some embodiments, methods are provided for treating animals having a disease or disorder that would benefit from altering the gene level, the method comprising administering to the animal a therapeutically effective amount of a pharmaceutical composition comprising (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol, wherein the gene is selected from one or more of ICA1, MRPS9, NECTIN3, EPHA3, MFF_DT, ROCK1P1, LYPD6, DKK2, CAP2, or GRM4.

[0101] In some embodiments, a method is provided for treating an animal having a disease or disorder, the method comprising the step of administering to the animal a therapeutically effective amount of a pharmaceutical composition comprising (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol, wherein the animal is selected by altered genes in cells that respond to treatment with (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol, and the genes are HS6ST2-AS1, CYP1B1, M EGF6, AHRR, CILP, TNS4, CACNA1S, IFNLR1, KSR2, OLFML2B, BCL11B, FGF18, GALNT5, NKAIN1, DAPP1, LINC00599, GPAT3, IL1B, PRAL, FXYD3, CYP26B1, CYP1 B1-AS1, CH25H, COLEC12, TIPARP, KIAA1549L, IFI30, NPTX1, ZMIZ1-AS1, ANPEP, ARHGEF16, WFDC11, KIAA1549, SUSD3, H19, GFRA1, NRG1, NQO1, TCF7, DTX 1, PRXL2A, COLGALT2, LUM, HSPB6, SLC16A10, COL24A1, KCND3, SLC1A2, WNT11, CLDN7, RNF128, PTGS1, ZNF385D, ADGRE2, LINC00639, ADGRD1, PRELP, TGF BI, FAM47E-STBD1, TSPAN15, SLC2A1, IER3, AJUBA, NTN4, CEMIP, CX3CL1, SLC16A3, STC2, THBD, IGFBP2, TNFRSF1B, ANXA3, NRP1, CD248, CDC42EP2, IGSF3 , ETNPPL, APCDD1, GNAZ, S1PR2, STK26, RDH10, RRAD, TNFAIP2, IL15RA, ITGB4, CD14, CAMK2B, SYNJ2, PIR, FAM156A, TPRA1, F2R, RGL1, GATAD2B, SLC6A6, N EDD4L, MN1, ATF3, ADCY9, SASH1, SLC22A23, SLC7A5, ANXA11, ZNF70, CD58, IL6R, ATXN1, FHL2, GABRA2, CSGALNACT1, S100A4, BIN1, PAPSS2, LMO4, RAB31,Select one or more from the following: EPM2A, TP53INP1, SESN3, LGI2, C14orf132, ENHO, GCSHP5, LRATD1, LGALS7B, MARCHF3, CRYAB, ID1, PLPP4, RPSAP58, EEF1A1P5, TPPP3, MBNL1-AS1, CCDC171, TMEM189-UBE2V1, AFF2, SCIN, ID3, RUBCNL, EFEMP1, MYRIP, MEST, MDFI, LY6D, RHBDL3, SAMD11, OR2S1P, HSPE1-MOB4, LGALS7, HPGD, SULT1E1, LINC00598, or ONECUT2.

[0102] In some embodiments, a method is provided for treating an animal having a disease or disorder, the method comprising administering to the animal a therapeutically effective amount of a pharmaceutical composition comprising (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol, wherein the animal is selected by altered genes that respond in cells to treatment with (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol, and the genes are selected from one or more of the following 14 genes: ICA1, MRPS9, NRIP3, NECTIN3, GATD3A, COL5A3, EPHA3, MFF_DT, IGFBP5, ROCK1P1, LYPD6, DKK2, CAP2, or GRM4.

[0103] In some embodiments, a method is provided for treating an animal having a disease or disorder, the method comprising administering to the animal a therapeutically effective amount of a pharmaceutical composition comprising (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol, wherein the animal is selected by altered genes that respond in cells to treatment with (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol, and the genes are selected from one or more of the following 14 genes: ICA1, MRPS9, NECTIN3, EPHA3, MFF_DT, ROCK1P1, LYPD6, DKK2, CAP2, or GRM4.

[0104] In some embodiments of the method, (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol is administered at a dose of 0.12 mg / kg or higher.

[0105] In some embodiments of the method, (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol is administered at a dose of 5 to 5000 mg / day.

[0106] In some embodiments of the method, (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol is administered parenterally, enterally, or topically.

[0107] In some embodiments of the method, 6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol is administered orally, sublingually, buccally, pulmonaryly, intranasally, intravenously, intramuscularly, or subcutaneously.

[0108] The use of (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol is provided for the preparation of a pharmaceutical for treating a human having the disease described in claim 40.

[0109] In some embodiments of the method, increasing, decreasing, or altering is determined against the corresponding measurement prior to the step of contacting the cells with (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol.

[0110] In some embodiments of the method, the cells are cells of a human subject, and the human subject has Alzheimer's disease, amyotrophic lateral sclerosis (ALS), Friedreich's ataxia, Huntington's disease (HD), or Parkinson's disease (PD).

[0111] In another embodiment, the Disclosure provides an in vitro method for screening therapeutic candidate(s) for their ability to treat animals, the method being: (1) Exposing animals to potential therapeutic agents; (2) Compare the levels of gene expression in cells before and after treatment after a certain period of time; (3) Based on the comparison results, including determining whether the animals can benefit from treatment with the candidate drug.

[0112] In one embodiment, the method may be an in vitro method.

[0113] In one embodiment, the period of exposure to cells is 1 to 30 days, preferably 1 to 15 days, and preferably 1 to 5 days.

[0114] In one embodiment, the candidate therapeutic agent is (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol.

[0115] In one embodiment, the animal is a mammal. In another embodiment, the mammal is a human or a non-human mammal. In a further embodiment, the mammal is a human.

[0116] In one embodiment, the cells in one of the above embodiments or in other embodiments herein are derived from a cell type or tissue selected from one or more of the following: adrenal gland, bone marrow, brain, breast, bronchi, caudate nucleus, cerebellum, cerebral cortex, cervix, uterus, colon, endometrium, epididymis, esophagus, fallopian tube, gallbladder, myocardium, hippocampus, kidney, liver, lung, lymph node, nasopharynx, oral mucosa, ovary, pancreas, parathyroid gland, peripheral blood mononuclear cells (PBMCs), placenta, prostate, rectum, salivary gland, seminal vesicle, skeletal muscle, skin, small intestine (including duodenum, jejunum, and ileum), smooth muscle, spleen, stomach, testis, thyroid gland, tonsil, bladder, and vagina. In further embodiments, the brain cells are derived from brain tissue selected from the cerebrum (including the cerebral cortex, basal ganglia (often called the striatum), and olfactory bulb), cerebellum (including the dentate nucleus, intermediate nucleus, fastigial nucleus, and vestibular nuclei), diencephalon (including the thalamus, hypothalamus, etc., and the posterior part of the pituitary gland), and brainstem (including the midbrain, pons, substantia nigra, and medulla oblongata). In further embodiments, the brain cells are selected from neurons or glial cells (e.g., astrocytes, oligodendrocytes, or microglia). In further embodiments, the cells are peripheral blood mononuclear cells (PBMCs).

[0117] In one embodiment, the cells are animal cells, for example, mammalian cells. In a further embodiment, the cells are human cells or non-human cells. In a further embodiment, the cells are in vitro, in vivo, or ex vivo.

[0118] In another embodiment, the cells are disease cells. In another embodiment, the cells are disease cells derived from a patient suffering from a disease or disorder as defined below.

[0119] In another embodiment, gene expression is HS6ST2-AS1, CYP1B1, MEGF6, AHRR, CILP, TNS4, CACNA1S, IFNLR1, KSR2, OLFML2B, BCL11B, FGF18, GALNT5, NKAIN1, DAPP1, LINC0059 9、GPAT3、IL1B、PRAL、FXYD3、CYP26B1、CYP1B1-AS1、CH25H、COLEC12、TIPARP、KIAA1549L、IFI30、NPTX1、ZMIZ1-AS1、ANPEP、ARHGEF16、WFDC11、KIAA1549L 9、SUSD3、H19、GFRA1、NRG1、NQO1、TCF7、DTX1、PRXL2A、COLGALT2、LUM、HSPB6、SLC16A10、COL24A1、KCND3、SLC1A2、WNT11、CLDN7、RNF128、PTGS1、ZNF38 5D、ADGRE2、LINC00639、ADGRD1、PRELP、TGFBI、FAM47E-STBD1、TSPAN15、SLC2A1、IER3、AJUBA、NTN4、CEMIP、CX3CL1、SLC16A3、STC2、THBD、IGFBP2、TNFR SF1B、ANXA3、NRP1、CD248、CDC42EP2、IGSF3、ETNPPL、APCDD1、GNAZ、S1PR2、STK26、RDH10、RRAD、TNFAIP2、IL15RA、ITGB4、CD14、CAMK2B、SYNJ2、PIR、FA M156A、TPRA1、F2R、RGL1、GATAD2B、SLC6A6、NEDD4L、MN1、ATF3、ADCY9、SASH1、SLC22A23、SLC7A5、ANXA11、ZNF70、CD58、IL6R、ATXN1、FHL2、GABRA2、CSGA LNACT1、S100A4、BIN1、PAPSS2、LMO4、RAB31、EPM2A、TP53INP1、SESN3、LGI2、C14orf132、ENHO、GCSHP5、LRATD1、LGALS7B、MARCHF3、CRYAB、ID1、PLPP4、R PSAP58、EEF1A1P5、TPPP3、MBNL1-AS1、CCDC171、TMEM189-UBE2V1、AFF2、SCIN、ID3、RUBCNL、EFEMP1、MYRIP、MEST、MDFI、LY6D、RHBDL3、SAMD11、OR2S1P、Select one or more from HSPE1-MOB4, LGALS7, HPGD, SULT1E1, LINC00598, and ONECUT2.

[0120] In another embodiment, the gene is selected from one or more of ICA1, MRPS9, NRIP3, NECTIN3, GATD3A, COL5A3, EPHA3, MFF_DT, IGFBP5, ROCK1P1, LYPD6, DKK2, CAP2, and GRM4.

[0121] In some embodiments, the following: A) Predicting the effectiveness of treatment for a disease or disorder in a subject, wherein the treatment comprises a first administration of an active pharmaceutical agent to a first cell of the subject, and optionally a second administration of the active pharmaceutical agent to a second cell of the subject, wherein the first cell has a first cell type, the second cell has a first cell type, and the active pharmaceutical agent is given by the following formula [ka] or predict having a pharmaceutically acceptable salt thereof; B) Collecting a first biological sample from the subject before the first administration (for example, less than one month before, for example less than one week before), wherein the control cells have a first cell type; C) After the first administration (e.g., up to 1, 2, 3, or 30 days later) and optionally before the second administration (e.g., less than one month prior, e.g., less than one week prior), the second biological sample is collected such that it contains the cells of the first administration; D) Determining the first differential expression of one or more genes in a second biological sample compared to a first biological sample; E) A method is provided which includes predicting the effectiveness of a treatment based on a first differential manifestation.

[0122] In some embodiments, the following: A) Monitoring the treatment of a disease or disorder in a subject, wherein the treatment includes a first administration of an active pharmaceutical agent to a first cell of the subject, and optionally a second administration of the active pharmaceutical agent to a second cell of the subject, wherein the first cell has a first cell type, the second cell has a first cell type, and the active pharmaceutical agent has the following formula [ka] or a pharmaceutically acceptable salt thereof, to be monitored; B) Collecting a first biological sample from the subject before the first administration (for example, less than one month before, for example less than one week before), wherein the control cells have a first cell type; C) After the first administration (e.g., up to 1, 2, 3, or 30 days later) and optionally before the second administration (e.g., less than one month prior, e.g., less than one week prior), the second biological sample is collected such that it contains the cells of the first administration; D) Determining the first differential expression of one or more genes in a second biological sample compared to a first biological sample; E) A method is provided which includes monitoring the treatment based on a first differential expression.

[0123] In some embodiments, the disease or disorder is a neurological disorder.

[0124] In some embodiments, the disease or disorder is Alzheimer's disease, amyotrophic lateral sclerosis (ALS), Friedreich's ataxia, Huntington's disease (HD), or Parkinson's disease (PD).

[0125] In some embodiments, the disease or disorder is age-related macular degeneration, atherosclerosis, autism spectrum disorder (ASD), benign focal muscular atrophy, cerebral infarction, Creutzfeldt-Jakob disease, Crohn's disease, Duchenne palsy, Friedreich's ataxia, frontotemporal dementia (FTD), glaucoma, hereditary spastic paraplegia (HSP), inclusion body myositis (IBM), inflammatory bowel disease, ischemia, Kugelberg-Wellander syndrome, Lewy body disease (LBD), multiple sclerosis (MS), myocardial infarction, necrotizing enterocolitis, neurofibromatosis type 1, Paget's disease of bone (PDB), primary lateral sclerosis (PLS), progressive bulbar palsy (PBP), progressive muscular atrophy The following conditions may be selected from: PMA (Progressive Malocclusion), pseudobulbar palsy, spinal muscular atrophy (SMA), ulcerative colitis, balocin-containing protein (VCP) related disorders, or one or more of the following: Werdnig-Hoffmann disease, transient ischemic attack, ischemia, cerebral hemorrhage, senile cataract, retinal ischemia, retinal vasculitis, Brown-Bearet-van Rare syndrome, Eels disease, meningitis and encephalitis, post-traumatic stress disorder, Charcot-Marie-Tooth disease, macular degeneration, hereditary Leber's optic atrophy (Kennedy disease), presenile dementia, depressive disorder, temporal lobe epilepsy, hereditary Leber's optic atrophy, cerebrovascular disease, subarachnoid hemorrhage, or schizophrenia.

[0126] In some embodiments, one or more genes are HS6ST2-AS1, CYP1B1, MEGF6, AHRR, CILP, TNS4, CACNA1S, IFNLR1, KSR2, OLFML2B, BCL11B, FGF18, GALNT5, NKAIN1, DAPP1, LINC00599, GPAT3, IL1B, PRAL, FXYD3, CYP26B1, CYP1B1-AS1, CH25H, COLEC12, TIPARP, KIAA1549L, IFI30, NPTX1, ZMIZ1-AS1, ANPEP, ARHGEF16, WFDC 11, KIAA1549, SUSD3, H19, GFRA1, NRG1, NQO1, TCF7, DTX1, PRXL2A, COLGALT2, LUM, HSPB6, SLC16A10, COL24A1, KCND3, SLC1A2, WNT11, CLDN7, RNF128, PT GS1, ZNF385D, ADGRE2, LINC00639, ADGRD1, PRELP, TGFBI, FAM47E-STBD1, TSPAN15, SLC2A1, IER3, AJUBA, NTN4, CEMIP, CX3CL1, SLC16A3, STC2, THBD, IG FBP2, TNFRSF1B, ANXA3, NRP1, CD248, CDC42EP2, IGSF3, ETNPPL, APCDD1, GNAZ, S1PR2, STK26, RDH10, RRAD, TNFAIP2, IL15RA, ITGB4, CD14, CAMK2B, SYNJ 2, PIR, FAM156A, TPRA1, F2R, RGL1, GATAD2B, SLC6A6, NEDD4L, MN1, ATF3, ADCY9, SASH1, SLC22A23, SLC7A5, ANXA11, ZNF70, CD58, IL6R, ATXN1, FHL2, GAB RA2, CSGALNACT1, S100A4, BIN1, PAPSS2, LMO4, RAB31, EPM2A, TP53INP1, SESN3, LGI2, C14orf132, ENHO, GCSHP5, LRATD1, LGALS7B, MARCHF3, CRYAB, ID1 , PLPP4, RPSAP58, EEF1A1P5, TPPP3, MBNL1-AS1, CCDC171, TMEM189-UBE2V1, AFF2, SCIN, ID3, RUBCNL, EFEMP1, MYRIP, MEST, MDFI, LY6D, RHBDL3, SAMD11,Select from OR2S1P, HSPE1-MOB4, LGALS7, HPGD, SULT1E1, LINC00598, or ONECUT2.

[0127] In some embodiments, one or more genes are selected from ICA1, MRPS9, NRIP3, NECTIN3, GATD3A, COL5A3, EPHA3, MFF_DT, IGFBP5, ROCK1P1, LYPD6, DKK2, CAP2, or GRM4.

[0128] In some embodiments, one or more genes are selected from ICA1, MRPS9, NECTIN3, EPHA3, MFF_DT, ROCK1P1, LYPD6, DKK2, CAP2, or GRM4.

[0129] In some embodiments, the first differential expression of one or more genes identifies significantly dysregulated transcripts of one or more genes in the first biological sample compared to a second biological sample.

[0130] In some embodiments, the method is an in vitro method.

[0131] In some embodiments, the first cell type is derived from one or more tissues selected from the adrenal gland, bone marrow, brain, breast, bronchi, caudate nucleus, cerebellum, cerebral cortex, cervix, uterus, colon, endometrium, epididymis, esophagus, fallopian tube, gallbladder, myocardium, hippocampus, kidney, liver, lung, lymph node, nasopharynx, oral mucosa, ovary, pancreas, parathyroid gland, peripheral blood mononuclear cells (PBMCs), placenta, prostate, rectum, salivary gland, seminal vesicle, skeletal muscle, skin, small intestine (including duodenum, jejunum, and ileum), smooth muscle, spleen, stomach, testis, thyroid gland, tonsil, bladder, or vagina.

[0132] In some embodiments, the animal is a mammal. In some embodiments, the mammal is a non-human animal. In some embodiments, the mammal is a human.

[0133] In some embodiments, the active pharmaceutical agent is administered at a dose of 0.12 mg / kg or higher. In some embodiments, the active pharmaceutical agent is administered at a dose of 5 to 5000 mg / day. In some embodiments, the active pharmaceutical agent is administered parenterally, enterally, or topically. In some embodiments, the active pharmaceutical agent is administered orally, sublingually, buccally, pulmonaryly, intranasally, intravenously, intramuscularly, or subcutaneously.

[0134] In some embodiments, the method is F) Monitoring the effectiveness of the treatment based on a second differential manifestation or a third differential manifestation; G) Continue treatment with the second dose; H) After the second administration, a third biological sample is collected from the subject, the third biological sample containing the second cells, I) further comprising determining the second differential expression of one or more genes in comparison to a first biological sample or in comparison to a third biological sample.

[0135] In some embodiments, subjects experience an increase in p62 levels, an increase in LC3 levels, or a decrease in mitochondrial morphological factors as a result of a first dose, a second dose, or both.

[0136] In some embodiments, the first cell type is a brain cell derived from brain tissue selected from the cerebrum, cerebellum, diencephalon, or brainstem, and optionally, the brain cell is an induced brain cell.

[0137] In some embodiments, the first cell type is a brain cell selected from neurons, astrocytes, oligodendrocytes, or microglia, and optionally, the brain cell is an induced brain cell.

[0138] In some embodiments, the first cell type is a brain cell selected from sensory neurons, motor neurons, interneurons, or cerebral neurons, and optionally, the brain cell is an induced brain cell.

[0139] In some embodiments, the method is a method for treating a disease or disorder.

[0140] A method is provided for treating Alzheimer's disease, amyotrophic lateral sclerosis (ALS), Friedreich's ataxia, Huntington's disease (HD), or Parkinson's disease (PD) in subjects requiring such treatment, and the method is 1) At a first dose of at least 0.12 mg / kg, and optionally at a dose of 5 to 5000 mg / day, the following formula [ka] Or administering an active pharmaceutical agent having a pharmaceutically acceptable salt thereof to a subject parenterally, enterally, or topically; 2) Measuring the levels of p62, LC3, or mitochondrial morphological factors in a biological sample containing cells, wherein the cells may include brain cells derived from brain tissue selected from the cerebrum, cerebellum, diencephalon, or brainstem, and optionally the brain cells are induced brain cells, wherein the measurement includes translatome profiling; 3) If the subject does not experience an increase in p62 levels, an increase in LC3 levels, or a decrease in mitochondrial morphological factors as a result of administration of the first dose, the treatment of Alzheimer's disease, amyotrophic lateral sclerosis (ALS), ataxia Friedreich, Huntington's disease (HD), or Parkinson's disease (PD) in the subject with a second dose greater than the first dose (e.g., a second dose greater than 0.12 mg / kg, and optionally, a dose from greater than 5 mg / day to about 5000 mg / day).

[0141] In some embodiments of the methods described herein, a measurement, determination, or sampling step, or a step similarly referred to herein, is performed up to 1, 2, 3, or 30 days after an administration step (e.g., a first or second administration of the API) and / or less than 1 month before an administration step (e.g., a first or second administration of the API), or less than 5, 4, 3, 2, or 1 week before an administration step(s) (e.g., a first or second administration of the API). In some embodiments of the methods described herein, administration of the API may refer to contacting cells (which may be isolated cells (e.g., in vitro or ex vivo) or cells from the body of the subject) with the API.

[0142] The pharmaceutical compositions disclosed herein comprise a therapeutically effective amount of (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol and at least one pharmaceutically acceptable excipient. The term “excipient” refers to a pharmaceutically acceptable inert substance used as a carrier for the pharmaceutically active ingredient ((6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol), and includes antifouling agents, binders, coating agents, disintegrants, fillers, diluents, solvents, flavoring agents, bulking agents, colorants, flow enhancers, dispersants, wetting agents, lubricants, preservatives, adsorbents, and sweeteners. The choice of excipient(s) depends on factors such as the nature of the specific mode of administration and dosage form. The solution or suspension used for injection or infusion may contain the following components: sterile diluents, e.g., water for injection, physiological saline, fixative oil, polyethylene glycol, glycerin, propylene glycol, or other synthetic solvents; antimicrobial agents, e.g., benzyl alcohol or methylparaben; antioxidants, e.g., ascorbic acid or sodium bisulfite; chelating agents, e.g., ethylenediaminetetraacetic acid; buffers, e.g., acetates, citrates, or phosphates; and agents for adjusting tonicity, e.g., sodium chloride or dextrose. The pH may be adjusted with an acid or base, e.g., hydrochloric acid or sodium hydroxide. Parenteral preparations may be sealed in disposable syringes such as ampoules or autoinjectors, or in multi-dose vials made of glass or plastic.

[0143] The pharmaceutical formulations of this disclosure may be any pharmaceutical dosage form. The pharmaceutical formulations may be, for example, tablets, capsules, nanoparticle materials, such as granulated particle materials or powders, lyophilized products for reconstitution, liquid solutions, suspensions, emulsions or other liquid forms, injectable suspensions, solutions, emulsions, etc., suppositories, or topical or transdermal formulations or patches. The pharmaceutical formulations generally contain about 1% to about 99% by weight of (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol, and 99% to 1% by weight of preferred pharmaceutical excipients. In one embodiment, the dosage form is an oral dosage form. In another embodiment, the dosage form is a parenteral dosage form. In another embodiment, the dosage form is an enteral dosage form. In another embodiment, the dosage form is a topical dosage form. In one embodiment, the pharmaceutical dosage form is a unit dose. The term "unit dose" refers to the amount of (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol administered to a patient as a single dose.

[0144] In some embodiments, the pharmaceutical compositions of the present disclosure are delivered to a subject via parenteral, enteral, or topical routes.

[0145] Examples of parenteral routes in this disclosure include, without limitation, one or more of the following: intraabdomen, intraamniotic, intraarterial, intraarticular, intrabiliary, intrabronchial, intrabursal, intracardiac, intracartilage, intracavitary, intracavernosus, intracavitary, intracerebral, intracisional, intracerebral, intradural, intraepidermal, intraesophageal, intragastric, intracranial, intradermal, intradiscal, intratubular, intraduodenal, intradural, intraepidermal, intraesophageal, intragastric, intraileal, intralesional, intratubular, intralymphatic, intramedullary, intrameningeal, intramuscular, intraocular, intraovarian, intrapericardial, intraperitoneal, intrapleural, intraprostatic, intrapulmonary, intraocular, intrasinusial, intraspinal, intrasynovial, intratendinous, intratendinous, intratesticular, intramedullary, intrathoracic, and / or subcutaneous.

[0146] The enteral routes of administration described herein include administration to the gastrointestinal tract via the mouth (oral), stomach (intragastric), and rectum (rectal). Gastric administration typically involves the use of a tube inserted through the nasal cavity (NG tube) or a tube inserted directly into the stomach (PEG tube). Rectal administration typically involves rectal suppositories. Oral administration includes sublingual and buccal administration.

[0147] Topical administration includes administration to body surfaces such as skin or mucous membranes, and also includes intranasal and pulmonary administration. Transdermal forms include creams, foams, gels, lotions, or ointments. Intranasal and pulmonary forms include liquids and powders, such as liquid sprays.

[0148] Dosage may vary depending on the dosage form used, patient sensitivity, and route of administration. Dosage and administration are adjusted to provide a sufficient level of the active ingredient or to maintain the desired effect. Factors to consider include the severity of the disease state, the subject's general health, the subject's age, weight, and sex, diet, timing and frequency of administration, concomitant drug use, response sensitivity, and tolerance / response to therapy.

[0149] In one embodiment, the daily dose of (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol administered to a patient is selected from the following: up to 200 mg, 175 mg, 150 mg, 125 mg, 100 mg, 90 mg, 80 mg, 70 mg, 60 mg, 50 mg, 30 mg, 25 mg, 20 mg, 15 mg, 14 mg, 13 mg, 12 mg, 11 mg, 10 mg, 9 mg, 8 mg, 7 mg, 6 mg, 5 mg, 4 mg, 3 mg, or up to 2 mg. In another embodiment, the daily dose is at least 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, 10 mg, 12 mg, 13 mg, 14 mg, 15 mg, 20 mg, 25 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 700 mg, 800 mg, 900 mg, 1,000 mg, 2,000 mg, 3,000 mg, 4,000 mg, or at least 5,000 mg. In another embodiment, the daily dose is 1-2 mg, 2-4 mg, 1-5 mg, 5-7.5 mg, 7.5-10 mg, 10-15 mg, 10-12.5 mg, 12.5-15 mg, 15-17.7 mg, 17.5-20 mg, 20-25 mg, 20-22.5 mg, 22.5-25 mg, 25-30 mg, 25-27.5 mg, 27.5-30 mg, 30-35 mg, 35-40 mg, 40-45 mg g, or 45-50 mg, 50-75 mg, 75-100 mg, 100-125 mg, 125-150 mg, 150-175 mg, 175-200 mg, 5-200 mg, 5-300 mg, 5-400 mg, 5-500 mg, 5-600 mg, 5-700 mg, 5-800 mg, 5-900 mg, 5-1,000 mg, 5-2,000 mg, 5-5,000 mg, or more than 5,000 mg.

[0150] In another embodiment, the single dose of (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol administered to the patient is selected from the following: 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, 10 mg, 12 mg, 13 mg, 14 mg, 15 mg, 16 mg, 17 mg, 18 mg, 19 mg, 20 mg, 21 mg, 22 mg, 23 mg, 24 mg, 25 mg, 26 mg, 27 mg, 28 mg, 29 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 15 0mg, 160mg, 170mg, 180mg, 190mg, 200mg, 210mg, 220mg, 230mg, 240mg, 250mg, 260mg, 270mg, 280mg, 290mg, 300mg, 310mg, 320mg, 330mg, 340mg, 350mg, 360mg, 370mg, 380mg, 390mg, 400mg, 410mg, 420mg, 430mg, 440mg, 450mg, 460mg, 470mg, 480mg, 490mg, 500mg, 600mg, 700mg, 800mg, 900mg, 1,000mg, 2,000mg, 3,000mg, 4,000mg, or 5,000mg.In another embodiment, the single dose of (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol administered to the patient is selected from the following: 1-2 mg, 2-4 mg, 1-5 mg, 5-7.5 mg, 7.5-10 mg, 10-15 mg, 10-12.5 mg, 12.5-15 mg, 15-17.7 mg, 17.5-20 mg, 20-25 mg, 20-22.5 mg, 22.5-25 mg, 25-30 mg, 25-27.5 mg, 27.5-30 mg, 30-35 mg, 35-40 mg, 40-45 mg g, 45-50 mg, 50-75 mg, 75-100 mg, 100-125 mg, 125-150 mg, 150-175 mg, 175-200 mg, 200-225 mg, 225-250 mg, 250-275 mg, 275-300 mg, 300-325 mg, 325-350 mg, 350-375 mg, 375-400 mg, 400-425 mg, 425-450 mg, 450-475 mg, 475-500 mg, 500-1,000 mg, 1,000-2,000 mg, 3,000-4,000 mg, 4,000-5,000 mg, or over 5,000 mg. In one embodiment, a single dose is administered by a route selected from oral, buccal, or sublingual administration. In another embodiment, a single dose is administered by injection, for example, subcutaneous, intramuscular, or intravenous. In yet another embodiment, a single dose is administered by inhalation or intranasal administration.

[0151] As a non-limiting example, the dose of (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol administered by subcutaneous injection may be about 3 to 5,000 mg per day, administered in divided doses. The single dose of (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol administered by subcutaneous injection may be about 1 to 6 mg, preferably about 1 to 4 mg, 1 to 3 mg, or 2 mg. Other embodiments include a range of about 5 to 5,000 mg, preferably about 100 to 1,000 mg, 100 to 500 mg, 200 to 400 mg, 250 to 350 mg, or 300 mg. Subcutaneous injection may be preferred in patients requiring the injection to be divided into more than 10 doses per day. The continuous subcutaneous infusion dose may be 1 mg / hour daily and is generally increased up to a maximum of 4 mg / hour depending on the response.

[0152] The particulate dose of (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol administered by pulmonary administration, for example by inhalation using a pressurized metered-dose inhaler (pMDI), dry powder inhaler (DPI), soft mist inhaler, nebulizer, or other device, may be in the range of about 0.5 to 15 mg, preferably about 0.5 to 8 mg or 2 to 6 mg. Other embodiments include the range of about 5 to 5,000 mg, preferably about 100 to 1,000 mg, 100 to 500 mg, 200 to 400 mg, 250 to 350 mg, or 300 mg. The nominal dose (ND) of (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol administered by pulmonary administration, i.e., the amount of drug measured in a container (also known as the quantitative dose), may be in the range of, for example, 0.5–15 mg, 3–10 mg, 10–15 mg, 10–12.5 mg, 12.5–15 mg, 15–17.7 mg, 17.5–20 mg, 20–25 mg, 20–22.5 mg, 22.5–25 mg, 25–30 mg, 25–27.5 mg, 27.5–30 mg, 30–35 mg, 35–40 mg, 40–45 mg, or 45–50 mg. Other embodiments include doses ranging from about 5 to 5,000 mg, preferably about 100 to 1,000 mg, 100 to 500 mg, 200 to 400 mg, 250 to 350 mg, or 300 mg. Long-acting pharmaceutical compositions may be administered 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 times or more (preferably ≤10 times per day), every other day, every 3 to 4 days, weekly, or every two weeks, depending on the half-life and clearance rate of the particular formulation.

[0153] Examples The present invention should not be limited in scope by the specific embodiments described herein. In fact, various modifications of the invention, in addition to those described herein, will be apparent to those skilled in the art from the foregoing description and the accompanying drawings. Such modifications are intended to be included within the scope of the appended claims.

[0154] Furthermore, it should be understood that all values ​​are approximate and provided for illustrative purposes only. All references cited and discussed herein are incorporated herein by reference in whole to the same extent as if each reference were incorporated by reference individually. Example 1: In vitro cell model derived from fibroblasts of ALS patients

[0155] Over the past decade, in vitro modeling of neurodegeneration has seen remarkable advancements, primarily driven by the reprogramming of adult human fibroblasts into induced pluripotent stem cells (iPSCs) and induced neural progenitor cells (iNPCs). In the field of ALS research, this offers an opportunity to model familial and sporadic diseases in vitro.

[0156] NPCs (Numerical Progenitor Cells) extracted from the spinal cord of postmortem ALS patients are already successfully differentiated into motor neurons, astrocytes, and oligodendrocytes. Using this method to derive astrocytes avoids inducing major epigenetic changes. However, the availability of postmortem samples is limited. In addition, the drawbacks of reprogramming astrocytes from human-derived iPSCs include time-consuming protocols and the complex and highly variable maturation time of astrocytes.

[0157] Therefore, a promising alternative to iPSC resources is the direct reprogramming of fibroblasts from immunocompatible hosts into astrocytes. Instead of generating iPSCs, direct reprogramming involves the use of cell lineage transcription factors to convert adult somatic cells into a different cell type. This technique has been used to generate subspecific nervous systems, such as cholinergic, dopaminergic, and motor neurons. Direct reprogramming techniques have also been used to induce astrocytes from ALS patient fibroblasts, generating tripotent iNPCs from ALS patients and controls within one month. When these cells differentiated into astrocytes, they exhibited similar toxicity to motor neurons as autopsy-derived astrocytes in co-culture, making them a useful tool in the development of drug screening (Figure 1).

[0158] Methodology: iNPCs were generated from adult human fibroblasts derived from patients diagnosed with ALS and age-matched healthy controls using previously reported approaches (Kim et al., PNAS, 2001.108(19), 7838-7843; Meyer et al., PNAS, 2014.111(2), 829-832). iNPCs were differentiated into iAstrocytes by culturing the progenitor cells in iAstrocyte medium for a total of 7 days, with a medium change on day 3.

[0159] Induced astrocytes from control or ALS patients were used in co-culture assays to determine their effects on mouse motor neuron (MN) survival. Mouse embryonic stem cell-derived motor neurons expressing green fluorescent protein (GFP) under the control of the HB9 promoter were selected and added to patient and control-derived iAstrocytes. Meanwhile, androgravold, (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol, monomethyl fumarate (MMF), and riluzole were screened in this co-culture system of patient iAstrocytes and wild-type mouse MNs. Mouse MN survival was monitored on days 1 and 3 using confocal imaging acquisition.

[0160] result: MN survival on day 3 was assessed as the percentage of surviving MN cells observed on day 1. As expected, iAstrocytes from healthy controls did not significantly alter the survival of mouse MNs on day 3. The introduction of all four drugs also did not alter the survival of mouse MNs (Figures 2A-2B and 3A-3B).

[0161] When iAstrocytes from three ALS patients with the C9orf72 mutation (i.e., patients C9orf72_183, C9orf72_201, and C9orf72_78) were co-cultured with mouse MN cells, less than 33% of MN cells survived on day 3 among all three ALS patients. However, day 3 MN cell survival was significantly improved when androgravoride, (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol, and MMF were introduced into the culture. More specifically, (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol improved MN survival by up to 38%.

[0162] When iAstrocytes from ALS patients with SOD1 mutations (i.e., patient SOD1_210, SOD1_102, and SOD1_100) were co-cultured with mouse MN cells, approximately 40% or less of the MN cells survived on day 3. MN cell survival on day 3 was most significantly improved by the introduction of (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol.

[0163] When iAstrocytes from three ALS patients with sporadic ALS mutations (i.e., patients sALS_17, sALS_12, and sALS_009) were co-cultured with mouse MN cells, the survival of MN cells on day 3 varied between 21% and 40%. In this study, the survival of MN cells on day 3 was most significantly improved in the presence of androglafold (Figures 2A–2B, 3A–3B, and 4).

[0164] Example 2: Autophagy regulatory factor expression - p62 and LC3 proteins Immunocytochemistry was used to detect the percentage of cells positive for cytoplasmic p62 and the number of perinuclear p62 spots within the cytoplasm. In this staining, all ALS patient cells had a higher percentage of p62-positive cells and a higher number of perinuclear p62 spots than control iAstrocytes. Therefore, we investigated whether treatment with (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol, androglavold, and MMF, or riluzole affected the presence of p62 using the same protocol.

[0165] Immunocytochemical imaging showed that treatment with (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol and androgravoride increased p62 expression in control and all patient iAstrocyte lines, specifically in the perinuclear region of cells (Figure 5). When these images were quantified in terms of the percentage of p62-positive cells, there was a significant increase in the number of cells expressing the p62 protein 24 hours after treatment with (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol across control and patient cell lines (two-way ANOVA, multiple comparisons, n=3, p<0.05). Addition of MMF had no significant effect on p62 expression levels in control or patient iAstrocytes (Figures 6A-6C).

[0166] Similar to the percentage of p62-positive cells, treatment with (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol also resulted in a significant increase in the number of perinuclear p62 spots across control and patient iAstrocyte lines; the highest increase was observed in sALS iAstrocytes (two-way ANOVA, multiple comparisons, n=3, p<0.05). In contrast, MMF treatment had little effect on perinuclear p62 spots (Figures 7A-7C). When control and patient iAstrocytes were treated with riluzole, there was little change in both the percentage of p62-positive cells and the number of perinuclear p62 spots (Figures 8A-8B).

[0167] This increase in p62 expression after treatment with (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol and andrographoride may be explained by activation of the autophagy pathway. Therefore, we investigated the effects of these two compounds on the protein expression of the autophagy markers LC3-I / LC3-II. Since autophagy deficiency is commonly reported in C9ORF72-ALS, we focused on this genetic subgroup of the cell line.

[0168] Similar to the p62 staining results, Western blotting confirmed higher levels of p62 protein in the C9ORF72 iAstrocyte line compared to controls, and these protein levels were further increased after treatment with (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol and androglavold (Figures 9A-9C). The LC3 protein is responsible for the formation of autophagosomes (vesicles that carry unwanted proteins to be sent for degradation). It exists in two forms: LC3-I, located in the cytoplasm, and LC3-II, bound to the autophagosome membrane. The conversion from LC3-I to LC3-II suggests the initiation of autophagosome formation and is therefore a useful biomarker for detecting autophagy. C9ORF72 iAstrocytes presented increased levels of LC3-I protein compared to control cell lines (Figure 9D). This is a known mechanism associated with C9ORF72 haploinsufficiency, leading to the initial activation of autophagy, which is then affected by defective interactions of the C9ORF72-encoded protein with the Rab1 and ULK1 complex, thus inhibiting autophagosome formation and allowing for the accumulation of LC3-I in the cytoplasm. However, treatment with (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol or androgravoride had no effect on LC3-I or LC3-II levels in controls and C9ORF72 iAstrocytes (Figures 9E-9F).

[0169] Example 3: Mitochondrial Dynamics Mitochondria were labeled and visualized using fluorescent TMRM dyes to investigate mitochondrial morphology in patient iAstrocytes. Patient iAstrocytes demonstrated differences in mitochondrial dynamics compared to control strains; extensive mitochondrial fragmentation was observed in SOD1 ND29505, and all patient strains had a significantly higher percentage of perinuclear mitochondria compared to controls. Subsequently, we investigated whether the addition of antioxidants or riluzole affected changes in cellular mitochondrial dynamics.

[0170] Mitochondrial staining images showed intense fusion of the mitochondrial network after androglafolide treatment; the mitochondrial network of sALS17 after treatment with the compound appeared to form thin, hair-like structures around the network (Figure 10).

[0171] When stained images were quantified, there was a significant decrease in mitochondrial morphological factors associated with androglavolide treatment, indicating greater mitochondrial network fusion and fewer mitochondrial branchings (two-way ANOVA, multiple comparisons, n=3, p<0.0001). This network fusion was reflected by a significant increase in mitochondrial area after androglavolide treatment (two-way ANOVA, multiple comparisons, n=3, p<0.001). However, there was no significant difference in the percentage of perinuclear mitochondria after androglavolide treatment, indicating that the treatment did not affect the localization of mitochondria within the network.

[0172] Mitochondrial branching was also affected by riluzole and other antioxidant compounds; all compounds tested showed a variable significance decrease in mitochondrial morphological factors (Figures 11A-C, two-way ANOVA, multiple comparisons, n=3, Figure 11A: (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol p<0.001; Figure 11B: MMF p<0.05; and Figure 11C: riluzole p<0.01).

[0173] Example 4: RNA sequencing to determine the mechanism of action of antioxidant compounds RNA extraction and quality control The mechanisms of action of therapeutic agents were investigated to understand whether they could produce a beneficial reduction in astrocyte toxicity in specific patient subgroups. Therefore, control and patient iAstrocytes were seeded as monocultures in 10 cm dishes and treated with compounds diluted in 0.01% DMSO for 48 hours. Cells were lysed, and RNA was extracted using a translatome protocol.

[0174] The quality of RNA samples, as well as the detection of any residual ribosomal RNA (rRNA), was assessed on a Picochip in an Agilent 2100 bioanalyzer, and sample volume was assessed using a Nanodrop system. Electropherograms were used to assess whether the sample could be advanced for sequencing, required separate collection for RNA degradation, or required purification from contaminated rRNA.

[0175] Transcriptional changes in patient astrocytes driven by (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol treatment

[0176] To determine the mechanism of action of the drugs, we investigated gene expression changes driven by drug treatment in each patient subgroup. Interestingly, untreated and treated samples tended to cluster very closely on the PCA plot, with the exception of the C9ORF72 sample, which showed a large shift toward the control sample on the major axis after drug treatment. This suggests that (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol treatment induced different expression changes in C9ORF72 patients compared to other groups.

[0177] To investigate the mechanism of action of (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol in patient iAstrocytes, a list of differentially modified transcripts between each patient iAstrocyte subgroup treated with the compound and each patient iAstrocyte subgroup that was not treated was generated by performing the comparisons in the following table (Table 1): [Table 1]

[0178] Venn diagrams of covalent genes between control and patient cell lines after (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol treatment are shown in Figures 12A–12B. After (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol treatment, CTR, SOD1, and sALS iAstrocyte lines showed similar numbers of intrinsic DEGs (CTR = 204 DEGs, 119 upregulations and 85 downregulations; SOD1 = 213 DEGs, 148 upregulations and 65 downregulations; and sALS = 202 DEGs, 47 upregulations and 155 downregulations). On the other hand, in C9ORF72 patient iAstrocytes, only 128 genes were significantly altered after treatment with (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol (64 elevated and 64 decreased). SOD1 and sALS strains shared the most genes (23 DEGs, 22 elevated and 1 decreased), but fewer genes were shared between different subgroups (9 DEGs, 6 elevated and 3 decreased); treatment with (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol resulted in transcriptional upregulation of the majority of genes shared between patient and control iAstrocytes.

[0179] When the original gene list was imported into the DAVID pathway analysis software, SOD1 iAstrocytes showed the highest number of significantly altered GO terms (44 pathways), followed by sALS (33 pathways), and CTR and C9ORF72 iAstrocytes (19 and 12 pathways, respectively). To identify drug-induced transcriptional changes common to all patient groups, we focused on pathways shared between two or more iAstrocyte groups. All shared pathways between control and patient iAstrocyte subgroups after (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol treatment are presented in Table 2. [Table 2]

[0180] Two pathways were shared between CTR and SOD1 iAstrocytes: the cellular response to organic cyclic compounds and the MAPK cascade. The first pathway refers to changes in cellular activity in response to organic cyclic compounds, including motility, secretion, enzyme production, or even further gene expression likely reflecting compound metabolism. This included significant upregulation in cytochrome P450 enzyme transcripts (CYP1A1:CTR p=1.87E-74, log2FC=+6.99, SOD1 p=2.18E-09, log2FC=+4.99; CYP1B1:CTR p=5.81E-06, log2FC=+2.62, SOD1 p=8.13E-27, log2FC=+2.59).

[0181] Treatment with (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol also induced DGE changes in transcripts within the MAP kinase signaling pathway, particularly in artemin (ARTN: CTR p=0.03, log2FC=+2.06, SOD1 p=0.004, log2FC=+2.93), a secretory ligand belonging to both the GDNF and TGF-β protein families, and interleukin-1 beta (IL1B: CTR p=4.36E-05, log2FC=+2.08, SOD1 p=1.21E-04, log2FC=+2.05), a key mediator of inflammatory responses.

[0182] However, there were many shared pathways among the patient iAstrocyte groups; two pathways across all patient groups, and six pathways between SOD1 and sALS iAstrocytes. This was interesting because treatment with (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol altered different pathways in control and patient iAstrocytes, with control strains showing minimal effects while patients exhibited significant gene expression changes. This reflected what had been observed in co-culture experiments and demonstrated a patient-specific drug response.

[0183] All patient groups presented DEGs associated with cell adhesion, but different transcripts were targeted within each group: contactin and protocadherin in C9ORF72, integrin in SOD1, and cadherin in sALS iAstrocytes. Compared to control iAstrocytes, all patient groups showed a significant increase in protocadherin-related transcripts, except in C9ORF72 iAstrocytes, where protocadherin transcripts decreased after (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol treatment (PCDHA4: p=7.90E-04, log2FC=-1.75; PCDHB15: p=0.02, log2FC=-2.02; PCDHGC4: p=0.003, log2FC=-3.06). While there was also a significant increase in CDH2 shared among all patient iAstrocytes, only the sALS strain demonstrated a decrease in cadherin transcripts after (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol treatment (CDH2: p=0.003, log2FC=-1.92; CDH6: p=7.51E-04, log2FC=-2.24). This suggests that (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol treatment may have an effect on cell migration and intercellular contact, but only in strains with a specific cellular environment.

[0184] It was not surprising that SOD1 and sALS iAstrocytes shared the most pathways, as they showed the highest number of shared transcripts after treatment with (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol. (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol appeared to target inflammatory responses within these cell types. SOD1 iAstrocytes showed downregulation of XC motif chemokine receptor 1 expression (XCR1: p=0.03, log2FC=-2.71) while exhibiting upregulation of IFN-induced T cell alpha chemotactic factors (CXCL11: p=0.02, log2FC=+2.70). On the other hand, sALS iAstrocyte showed downregulation of inflammatory chemotactic molecules (CXCL1: p-value = 0.003, log2FC = -3.22; CXCL6: p-value = 0.02, log2FC = -3.87).

[0185] Following treatment with (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol, there were also DGE changes in the immune response; in SOD1 iAstrocytes, there was increased expression of interleukin-1 alpha along with interleukin-1 beta (IL1A: p=0.03, log2FC=+2.53), while in sALS iAstrocytes, there was downregulation of interleukin-32 (IL32: p=0.03, log2FC=-2.07), and DGE in transcripts expressing major histocompatibility complex class II proteins (HLA-DOA: p=0.03, log2FC=+1.58; HLA-DQB1: p=0.005, log2FC=-3.85).

[0186] SOD1 and sALS iAstrocytes also shared DGE changes in the oxidative process after treatment, as expected from the known antioxidant properties of these drugs. Both groups showed significant upregulation in transcripts related to aldehyde dehydrogenases involved in the detoxification of long-chain aldehydes and the metabolism of neurotransmitters (ALDH3A1: SOD1 p=1.58E-11, log2FC=+2.09; sALS p=1.47E-11, log2FC=+1.85), which is related to NRF2. Interestingly, sALS iAstrocytes showed significant downregulation in another aldehyde dehydrogenase transcript involved in the synthesis of retinoic acid from retinal aldehyde (ALDH1A2: p=0.04, log2FC=-1.94), which is related to MN death in ALS. Cytochrome P450 enzymes, which are responsible for retinoic acid metabolism, were also differentially regulated in SOD1 and sALS iAstrocytes after treatment (CYP26B1:SOD1 p-value = 1.83E-05, log2FC = +1.50; CYP26A1:sALS p-value = 0.009, log2FC = -5.16).

[0187] Treatment with (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol significantly increased the expression of arachidonic acid 15-lipoxygenase, an essential factor for ferroptosis, in SOD1 iAstrocytes (ALOX15B: p-value = 3.19E-07, log2FC = +3.12), and resulted in increased expression of prostaglandin-endoperoxide synthase 1, which may be involved in iron binding (PTGS1: p-value = 4.49E-06, log2FC = +1.57), suggesting that the treatment may have an effect on iron levels in SOD1 iAstrocytes, as seen in spinal cord injury (SCI). sALS iAstrocytes show significant downregulation in the expression of peroxidacin, an enzyme that catalyzes peroxidation reactions using hydrogen peroxide produced by the NADPH oxidase enzyme (PXDN: p-value = 9.91E-05, log2FC = -3.57), which is a novel target of NRF2.

[0188] In summary, as initially hypothesized, treatment with (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol resulted in DGE changes specific to the patient group's mutational state. Mild changes in gene expression related to cytochrome P450 enzymes and MAPK signaling were shared between CTR and SOD1 patient strains. While all patient groups shared DGE changes in cell adhesion, only C9ORF72 iAstrocyte presented changes in protocadherin transcripts, and DGE in cadherin genes was present only in the sALS strain. The SOD1 and sALS strains showed the most DGE changes after treatment with (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol, and these were associated with pathways including inflammation, immune response, oxidation, retinoic acid metabolism, and iron binding.

[0189] Example 5: Baseline transcriptional ALS patient response In previous examples, RNA sequencing data were investigated to elucidate similarities and differences between patient subgroups (SOD1, C9ORF72, and sALS), as well as the mechanism of action of the drugs across the patient subgroups. In the current study, RNA sequencing samples were matched based on drug responses taken from co-culture data, as identified in Table 3 below. [Table 3]

[0190] Table 3 above shows that each patient response group has diverse genetic subgroups. Understanding whether the response group presented unique transcriptional features compared to the control is crucial as it will help identify what was unique for these patients. A list of DEGs for each patient iAstrocyte response group and the control iAstrocyte group was generated by performing the comparisons in Table 4. [Table 4]

[0191] Because this analysis investigated specific transcripts rather than pathways, DEGs were selected based on p-adj < 0.05, meaning that we anticipated that less than 5% of significant transcripts would be false positives.

[0192] DEGs specific to each patient response group were identified using multi-group comparisons with andrografolide and riluzole. There were 11 DEGs specific to patient strains that responded to (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol treatment (Table 5). These transcripts were involved in mitochondrial protein synthesis, cell adhesion, and receptor tyrosine kinase signaling.

[0193] Little is known about the MRPS9 gene in relation to ALS, but the effects of mutant SOD1 (mSOD1) on mitochondria have been widely reported. Deviations from normal SOD1 levels increased mitochondrial DNA (mtDNA) copy number, as well as impaired mitochondrial protein synthesis associated with mSOD1. Dipeptide repeat proteins (DPRs) associated with C9ORF72-ALS were also found to preferentially bind to mitochondrial ribosomal proteins and impair mitochondrial function. On the other hand, both SOD1 and C9ORF72 iAstrocytes showed significant reductions in differential mitochondrial dynamics and mitochondrial morphological factors after treatment with (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol.

[0194] Treatment with (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol has been shown to have an effect on transcripts related to cell adhesion. NECTIN3 encodes a cell adhesion molecule responsible for intercellular contact at adherens junctions, and its expression can be altered by stress. In neurons, these proteins are present at synaptic junctions, forming interneuronal connections and maintaining synapse formation and transmission. Nectin-3 expression has been reported to be decreased in Alzheimer's disease models in association with tauopathy, but the increased expression observed in iAstrocytes of ALS patients may be related to a stressed cellular environment. It was known that miR-29a expression was increased in SOD1 mouse models, and therefore this microRNA potentially regulates COL5A3 expression in iAstrocytes of SOD1 patients.

[0195] Ephrin receptors are the largest protein family of receptor tyrosine kinases and are responsible for intercellular interactions as well as nervous system development, specifically neuronal migration and axonal guidance. Loss of ephrin receptor signaling has been reported to be protective in ALS, while EPHA3 deletion was identified as a protective factor in the sALS patient population, and inhibition of Epha4 signaling increased survival in the SOD1 mouse model. This indicates that significant upregulation of EPHA3 in iAstrocytes of these ALS patients was detrimental to surrounding MNs.

[0196] Example 6: Response of a transcript to a drug in an ALS patient To identify transcripts that determine the “patient responder” specific response to treatment with (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol, altered gene expression after drug treatment was compared within patient responder cell lines. A list of differentially regulated transcripts between the (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol iAstrocyte response groups before and after treatment, compared with the control iAstrocyte group, was generated by performing the comparisons in the table below (Table 6). [Table 5] TIFF2026525465000010.tif162159 [Table 6]

[0197] The highest number of significant DEGs was observed in the (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol responder-treated (S Resp. T) vs. (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol responder-untreated (S Resp. U) comparison (161 genes), while other comparisons showed fewer transcripts; the S Resp. U vs. control / untreated (CTR U) comparison showed 44 DEGs, while the S Resp. T vs. CTR U comparison showed 73 genes. A complete list of transcripts generated from the S Resp. T vs. S Resp. U comparison is presented in Table 7. [Table 7] TIFF2026525465000013.tif226159

[0198] The gene list was imported into Venny to identify transcripts that were uniquely or commonly dysregulated in the three comparisons (Figure 13). There were 153 genes unique to the S Resp.T vs. S Resp.U comparison (111 elevated, 42 decreased). This comparison identified genes that showed significant changes in expression after treatment, potentially due to the mechanism of drug action. However, this list of 153 genes did not share any transcripts with the S Resp.U vs. CTRU comparison, meaning that none of these transcripts were dysregulated at baseline. [Table 8] TIFF2026525465000015.tif237159TIFF2026525465000016.tif240159TIFF2026525465000017.tif74159

[0199] Thirty-eight DEGs specific to the S Resp. T vs. CTR U comparison (28 upregulated, 10 downregulated) highlighted transcripts that significantly changed after treatment but did not counteract the original dysfunction in patient cell lines. This is because these transcripts did not change in the baseline comparison. DEGs shared between the S Resp. U vs. CTR U and S Resp. T vs. CTR U comparisons (27 transcripts, 27 upregulated, 0 downregulated) identified transcripts that were dysregulated at baseline and remained dysregulated after drug treatment without changing regulation, increase, or decrease. Therefore, these 14 genes specific to the S Resp. U vs. CTR U comparison (11 increases, 3 decreases) were further investigated. This is because these transcripts were significantly different from the control strain before treatment with (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol, but this significance was lost after treatment, thus indicating that (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol treatment corrected their dysregulation. Therefore, these transcripts would be good candidates for identifying response gene signatures.

[0200] To identify biomarkers that identified patient responders, we anticipated that gene expression biomarkers would be corrected after treatment. A baseline S Resp. U vs. CTR U comparison in previous examples served as a starting point, where these transcripts either 1) reversed direction and remained significantly dysregulated compared to controls, or 2) returned to baseline and were no longer significantly dysregulated after treatment with (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol.

[0201] Of the 14 genes (11 elevated, 3 decreased) that were specific to the baseline S Resp.U vs. CTRU comparison, 10 were independently dysregulated in M102 responders at baseline and corrected after treatment with (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol (Table 5).

[0202] Ten transcripts were specific to the S Resp. U vs. CTR U comparison, indicating that these transcripts were no longer significantly dysregulated in patient responder strains after treatment. Interestingly, ICA1, MRPS9, NECTIN3, EPHA3, MFF-DT, and ROCK1P1 were all previously investigated as significantly dysregulated transcripts specific to the (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol responder at baseline compared to other responder groups (Table 5). Further transcripts identified in the S Resp. U vs. CTR U comparison would likely have been significantly dysregulated in other response groups at baseline. The transcripts showing the greatest significant changes after treatment with (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol were MRPS9, MFF-DT, ROCK1P1, and GRM4 (Table 9). [Table 9]

[0203] To identify whether this change in significance after (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol treatment was due simply to the response in one or more patient-responder cell lines, normalized gene counts (TPM values) for each transcript before and after (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol treatment were plotted in both patient-responder and non-responder iAstrocytes (Figure 14). The graph showed that not all biomarkers changed in all responder cell lines after treatment. Therefore, it is possible that a combination of these transcripts could be used as a suitable biomarker or biomarker panel for drug response.

[0204] Next, patient cell lines were scored according to the changes in each biomarker after treatment with (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol (Figure 15). A score of +1 was given for changes toward the control after treatment, a score of 0 for no change, and a score of -1 for gene expression changes that were contrary to the control after treatment. The total scores are shown at the bottom of the table. The figure shows that when they were evaluated against the whole gene panel, patient responders were typically scored between 2 and 6, while patient non-responder lines were scored between 0 and -2. This confirms the previous hypothesis that although not all biomarkers changed in all patient lines, selected gene biomarkers can be used as a panel to distinguish between patient responders and non-responders.

[0205] Embodiment Embodiment 1 A method for increasing p62 protein in cells, comprising the step of contacting cells with an effective amount of (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol.

[0206] Embodiment 2: The method according to Embodiment 1, wherein the p62 protein is a perinuclear P62 protein.

[0207] Embodiment 3 A method for increasing LC3 protein in cells, comprising the step of contacting cells with an effective amount of (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol.

[0208] Embodiment 4 The method according to Embodiment 3, wherein the LC3 protein is LC3-I or LC3-II protein.

[0209] Embodiment 5 A method for reducing mitochondrial morphological factors in cells, comprising the step of contacting cells with an effective amount of (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol.

[0210] Embodiment 6 A method for altering a biological pathway in cells, comprising the step of contacting cells with an effective amount of (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol.

[0211] Embodiment 7 The method according to Embodiment 6, wherein the biological pathway is selected from one or more of the following: cellular response to organic cyclic compounds, MAPK cascade, epithelial fold morphogenesis, cell adhesion, angiogenesis, inflammatory response, immune response, redox process, xenobiotic metabolic process, cholinergic synaptic transmission, or collagen fiber organization.

[0212] Embodiment 8 A method for altering the level of gene expression in cells, comprising the step of contacting cells with an effective amount of (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol.

[0213] Embodiment 9 The genes are HS6ST2-AS1, CYP1B1, MEGF6, AHRR, CILP, TNS4, CACNA1S, IFNLR1, KSR2, OLFML2B, BCL11B, FGF18, GALNT5, NKAIN1, DAPP1, LINC00599, GPAT3, IL1B, PRAL, FXYD3, CYP26B1, CYP1B1-AS1, CH25H, COLEC12, TIPARP, KIAA1549L, IFI30, NPTX1, ZMIZ1-AS1, ANPEP, ARHGEF16, WFDC11, KIAA1549, SUSD3, H19, GFRA1, NRG1, NQO1, TCF7, DTX1, PRXL2A, COLGALT2, LUM, HSPB6, SLC16A10, COL24A1, KCND3, SLC1A2, WNT11, CLDN7, RNF128, PTGS1, ZNF385D, ADGRE2, LINC00639, ADGRD1, PRELP, TGFBI, FAM47E-STBD1, TSPAN15, SLC2A1, IER3, AJUBA, NTN4, CEMIP, CX3CL1, SLC16A3, STC2, THBD, IGFBP2, TNFRSF1B, ANXA3, NRP1, CD248, CDC42EP2, IGSF3, ETNPPL, APCDD1, GNAZ, S1PR2, STK26, RDH10, RRAD, TNFAIP2, IL15RA, ITGB4, CD14, CAMK2B, SYNJ2, PIR, FAM156A, TPRA1, F2R, RGL1, GATAD2B, SLC6A6, NEDD4L, MN1, ATF3, ADCY9, SASH1, SLC22A23, SLC7A5, ANXA11, ZNF70, CD58, IL6R, ATXN1, FHL2, GABRA2, CSGALNACT1, S100A4, BIN1, PAPSS2, LMO4, RAB31, EPM2A, TP53INP1, SESN3, LGI2, C14orf132, ENHO, GCSHP5, LRATD1, LGALS7B, MARCHF3, CRYAB, ID1, PLPP4, RPSAP58, EEF1A1P5, TPPP3, MBNL1-AS1, CCDC171, TMEM189-UBE2V1, AFF2, SCIN, ID3, RUBCNL, EFEMP1, MYRIP, MEST, MDFI, LY6D, RHBDL3, SAMD11, OR2S1P,The method according to Embodiment 8, selected from one or more of HSPE1-MOB4, LGALS7, HPGD, SULT1E1, LINC00598, or ONECUT2.

[0214] Embodiment 10 The method according to Embodiment 8, wherein the gene is selected from one or more of ICA1, MRPS9, NRIP3, NECTIN3, GATD3A, COL5A3, EPHA3, MFF_DT, IGFBP5, ROCK1P1, LYPD6, DKK2, CAP2, or GRM4.

[0215] Embodiment 11 The method according to any one of Embodiments 1 to 10, wherein the cells are derived from a cell type or tissue selected from one or more of the following: adrenal gland, bone marrow, brain, breast, bronchi, caudate nucleus, cerebellum, cerebral cortex, cervix, uterus, colon, endometrium, epididymis, esophagus, fallopian tube, gallbladder, myocardium, hippocampus, kidney, liver, lung, lymph node, nasopharynx, oral mucosa, ovary, pancreas, parathyroid gland, peripheral blood mononuclear cells (PBMCs), placenta, prostate, rectum, salivary gland, seminal vesicle, skeletal muscle, skin, small intestine (including duodenum, jejunum, and ileum), smooth muscle, spleen, stomach, testis, thyroid gland, tonsil, bladder, or vagina.

[0216] Embodiment 12 The method according to Embodiment 11, wherein the brain cells are derived from brain tissue selected from the cerebrum, cerebellum, diencephalon, or brainstem.

[0217] Embodiment 13 The method according to Embodiment 12, wherein the brain cells are selected from neurons, astrocytes, oligodendrocytes, or microglia.

[0218] Embodiment 14 The method according to Embodiment 13, wherein the neuron is a sensory neuron, a motor neuron, an interneuron, or a brain neuron.

[0219] Embodiment 15 The method according to any one of Embodiments 1 to 14, wherein the cells are animal cells.

[0220] Embodiment 16 The method according to Embodiment 15, wherein the cells are human cells.

[0221] Embodiment 17 The method according to any one of Embodiments 1 to 16, wherein the cells are in vitro.

[0222] Embodiment 18 The method according to any one of Embodiments 1 to 16, wherein the cells are ex vivo.

[0223] Embodiment 19 The method according to any one of Embodiments 1 to 16, wherein the cells are in vivo.

[0224] Embodiment 20 The method according to any one of Embodiments 1 to 16, wherein the cells are disease cells.

[0225] Embodiment 21 Disease cells are associated with age-related tau astrocytosis (ARTA), Alexander disease, Alzheimer's disease, amyotrophic lateral sclerosis (ALS), critical disease myopathy (CIM), primary age-related tauopathy (PART), aortic medial amyloidosis, ApoAI amyloidosis, ApoAII amyloidosis, ApoAIV amyloidosis, argyrophilic granulosis, ataxia telangiectasia, atrial fibrillation, autosomal dominant high IgE syndrome, atrial amyloidosis, Bloom syndrome, cardiovascular disease, coronary artery disease, myocardial infarction, stroke, restenosis, arteriosclerosis, cataracts, and brain disease. Amyloid angiopathy, Christianson syndrome, chronic traumatic encephalopathy, Cockayne syndrome, corneal lactoferrin amyloidosis, corticobasal degeneration, Crohn's disease, Cushing's disease, lichenoid amyloidosis, cystic fibrosis, dentatorubral-pallidoluysian atrophy (DRPLA), dialysis amyloidosis, diffuse neurofibrillary tangle disease with calcification, Down syndrome, endotoxin shock, Finnish familial amyloidosis, familial amyloid neuropathy, familial British dementia (FBD), familial Danish dementia (FDD), familial dementia Cognitive disorders, fibrinogen amyloidosis, fragile X syndrome, fragile X-associated tremor / ataxia syndrome (FXTAS), Friedreich's ataxia, frontotemporal degeneration, glaucoma, glycogen storage disease type IV (Andersen's disease), Guadeloupe-type parkinsonism, hereditary lattice corneal dystrophy, Huntington's disease, inclusion body myositis / myopathy, inflammation, inflammatory bowel disease, ischemic state, ischemia / reperfusion injury, myocardial ischemia, stable angina, unstable angina, stroke, ischemic heart disease and cerebral ischemia, light chain or heavy chain amyloidosis, lysosomal storage disease, aspartylglucosamineuria, Fabry disease, Batten disease, cystinosis, Faber disease, fucosidosis, galactosialidosis, Gaucher disease type 1, 2 or 3, GM1 gangliosidosis, Hunter disease, Haller-Scheye disease, Krabbe disease, α-mannosidosis, β-mannosidosis, Maroto-Lamy disease, metachromatic leukodystrophy, Morquio A syndrome, Morquio B syndrome, mucolipidosis type II, mucolipidosis type III, Niemann-Pick disease type A, B or C, Pompe disease, Sandhoff disease, Sanfilippo syndrome type A, B, C or D, Schindler disease, Schindler-Kanzaki disease,Sialidosis, Sly syndrome, Tay-Sachs disease, Wolmann disease, lysozyme amyloidosis, Mallory bodies, medullary thyroid carcinoma, mitochondrial myopathy, multiple sclerosis, multiple system atrophy, myotonic dystrophy, myotonic dystrophy, neurodegenerative diseases with cerebral iron deposition, neurofibromatosis, neuronal ceroid lipofuscinosis, odontogenic (Pinborg) tumor amyloid, Guam Parkinson's dementia complex, Parkinson's disease, peptic ulcer, Pick's disease, pituitary prolactinoma, post-encephalitis parkinsonism, prion disease (transmissible spongiform encephalopathy) (Creutzfeldt-Jakob disease (CJD), variant Creutzfeldt-Jakob disease, Gerstmann-Streussler-Scheinker syndrome, fatal familial The method according to Embodiment 20, which is derived from an animal having a disease or disorder selected from one or more of the following: sexual insomnia (including kuru), progressive supranuclear palsy, alveolar proteinosis, retinal ganglion cell degeneration in glaucoma, retinitis pigmentosa with rhodopsin mutations, seminal vesicle amyloidosis, senile systemic amyloidosis, serpinopathy, sickle cell disease, spinal and bulbar muscular atrophy (SBMA), spinocerebellar ataxia, spinocerebellar ataxia type 1, spinocerebellar ataxia type 2, spinocerebellar ataxia type 3 (Machado-Joseph disease), spinocerebellar ataxia type 6, spinocerebellar ataxia type 7, spinocerebellar ataxia type 8, spinocerebellar ataxia type 17), subacute sclerosing panencephalitis, tauopathy, type 2 diabetes mellitus, vascular dementia, or Werner syndrome.

[0226] Embodiment 22 A method for treating an animal having a disease or disorder which would benefit from increasing p62 protein, increasing LC3 protein, or decreasing mitochondrial morphological factors, or a combination thereof, the method comprising administering to the animal a therapeutically effective amount of a pharmaceutical composition comprising (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol.

[0227] Embodiment 23 A method for treating an animal having a disease or disorder which would benefit from altering a biological pathway in cells, the method comprising the step of contacting cells with an effective amount of (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol, wherein the biological pathway is selected from one or more of the following: cellular response to an organic cyclic compound, MAPK cascade, epithelial fold morphogenesis, cell adhesion, angiogenesis, inflammatory response, immune response, redox process, xenobiotic metabolic process, cholinergic synaptic transmission, or collagen fiber organization.

[0228] Embodiment 24 A method for treating an animal having a disease or disorder which would benefit from altering the level of a gene, the method comprising administering to the animal a therapeutically effective amount of a pharmaceutical composition comprising (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol, wherein the gene is HS6ST2-AS1, CYP1B1, MEGF6, AHRR, CILP, TNS4, CACNA1S, IFNLR1, KSR2, OLFML2B, BCL11B, FGF18, GALNT5, NKAIN1, DAPP1 , LINC00599, GPAT3, IL1B, PRAL, FXYD3, CYP26B1, CYP1B1-AS1, CH25H, COLEC12, TIPARP, KIAA1549L, IFI30, NPTX1, ZMIZ1-AS1, ANPEP, ARHGEF16, WFDC 11, KIAA1549, SUSD3, H19, GFRA1, NRG1, NQO1, TCF7, DTX1, PRXL2A, COLGALT2, LUM, HSPB6, SLC16A10, COL24A1, KCND3, SLC1A2, WNT11, CLDN7, RNF128, PT GS1, ZNF385D, ADGRE2, LINC00639, ADGRD1, PRELP, TGFBI, FAM47E-STBD1, TSPAN15, SLC2A1, IER3, AJUBA, NTN4, CEMIP, CX3CL1, SLC16A3, STC2, THBD, I GFBP2, TNFRSF1B, ANXA3, NRP1, CD248, CDC42EP2, IGSF3, ETNPPL, APCDD1, GNAZ, S1PR2, STK26, RDH10, RRAD, TNFAIP2, IL15RA, ITGB4, CD14, CAMK2B, SY NJ2, PIR, FAM156A, TPRA1, F2R, RGL1, GATAD2B, SLC6A6, NEDD4L, MN1, ATF3, ADCY9, SASH1, SLC22A23, SLC7A5, ANXA11, ZNF70, CD58, IL6R, ATXN1, FHL2, GABRA2, CSGALNACT1, S100A4, BIN1, PAPSS2, LMO4, RAB31, EPM2A, TP53INP1, SESN3, LGI2, C14orf132, ENHO, GCSHP5, LRATD1, LGALS7B, MARCHF3, CRYAB,A method selected from one or more of the following: ID1, PLPP4, RPSAP58, EEF1A1P5, TPPP3, MBNL1-AS1, CCDC171, TMEM189-UBE2V1, AFF2, SCIN, ID3, RUBCNL, EFEMP1, MYRIP, MEST, MDFI, LY6D, RHBDL3, SAMD11, OR2S1P, HSPE1-MOB4, LGALS7, HPGD, SULT1E1, LINC00598, or ONECUT2.

[0229] Embodiment 25 A method for treating an animal having a disease or disorder which would benefit from altering the level of a gene, the method comprising administering to the animal a therapeutically effective amount of a pharmaceutical composition comprising (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol, wherein the gene is selected from one or more of ICA1, MRPS9, NRIP3, NECTIN3, GATD3A, COL5A3, EPHA3, MFF_DT, IGFBP5, ROCK1P1, LYPD6, DKK2, CAP2, or GRM4.

[0230] Embodiment 26 A method for treating an animal having a disease or disorder which would benefit from altering the level of a gene, the method comprising administering to the animal a therapeutically effective amount of a pharmaceutical composition comprising (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol, wherein the gene is selected from one or more of ICA1, MRPS9, NECTIN3, EPHA3, MFF_DT, ROCK1P1, LYPD6, DKK2, CAP2, or GRM4.

[0231] Embodiment 27 The disease or disorder is age-related macular degeneration, Alzheimer's disease (AD), amyotrophic lateral sclerosis (ALS), atherosclerosis, autism spectrum disorder (ASD), benign focal muscular atrophy, cerebral infarction, Creutzfeldt-Jakob disease, Crohn's disease, Duchenne palsy, Friedreich's ataxia, frontotemporal dementia (FTD), glaucoma, hereditary spastic paraplegia (HSP), Huntington's disease (HD), inclusion body myositis (IBM), inflammatory bowel disease, ischemia, Kugelberg-Wellander syndrome, Lewy body disease (LBD), multiple sclerosis (MS), myocardial infarction, necrotizing enterocolitis, neurofibromatosis type 1, Paget's disease of bone (PDB), Parkinson's disease (PD), primary lateral sclerosis (PLS), progressive bulbar palsy (PBP), The method according to any one of embodiments 22 to 26, wherein the patient is one or more selected from among muscular atrophy (PMA), pseudobulbar palsy, spinal muscular atrophy (SMA), ulcerative colitis, balocin-containing protein (VCP) related disorder, or Werdnig-Hoffmann disease, transient ischemic attack, ischemia, cerebral hemorrhage, senile cataract, retinal ischemia, retinal vasculitis, Brown-Bearet-van Lehr syndrome, Eels disease, meningitis and encephalitis, post-traumatic stress disorder, Charcot-Marie-Tooth disease, macular degeneration, X-linked spinal muscular atrophy, presenile dementia, depressive disorder, temporal lobe epilepsy, hereditary Leber's optic nerve atrophy (Kennedy disease), cerebrovascular disease, subarachnoid hemorrhage, or schizophrenia.

[0232] Embodiment 28 The method according to any one of Embodiments 22 to 27, wherein the animal is a mammal.

[0233] Embodiment 29 The method according to Embodiment 28, wherein the mammal is a non-human animal.

[0234] Embodiment 30 The method according to Embodiment 28, wherein the mammal is a human.

[0235] Embodiment 31: The method according to any one of Embodiments 22 to 30, wherein (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol is administered in a dose of 0.12 mg / kg or more.

[0236] Embodiment 32: The method according to any one of Embodiments 22 to 30, wherein (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol is administered in a dose between 5 and 5000 mg / day.

[0237] Embodiment 33: The method according to any one of Embodiments 22 to 30, wherein (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol is administered parenterally, enterally, or topically.

[0238] Embodiment 34 The method according to Embodiment 33, wherein 6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol is administered orally, sublingually, buccally, pulmonaryly, intranasally, intravenously, intramuscularly, or subcutaneously.

[0239] Embodiment 35: Use of (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol for the preparation of a pharmaceutical product for treating a human having the disease described in Embodiment 27.

[0240] Embodiment 36 A method for treating an animal having a disease or disorder, the method comprising the step of administering to the animal a therapeutically effective amount of a pharmaceutical composition comprising (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol, wherein the animal is selected by altered genes in cells that respond to treatment with (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol, and the genes are HS6ST2-AS1, CYP1B1, MEGF6, AHRR, CIL P, TNS4, CACNA1S, IFNLR1, KSR2, OLFML2B, BCL11B, FGF18, GALNT5, NKAIN1, DAPP1, LINC00599, GPAT3, IL1B, PRAL, FXYD3, CYP26B1, CYP1B1-AS1, CH25 H, COLEC12, TIPARP, KIAA1549L, IFI30, NPTX1, ZMIZ1-AS1, ANPEP, ARHGEF16, WFDC11, KIAA1549, SUSD3, H19, GFRA1, NRG1, NQO1, TCF7, DTX1, PRXL2A, C OLGALT2, LUM, HSPB6, SLC16A10, COL24A1, KCND3, SLC1A2, WNT11, CLDN7, RNF128, PTGS1, ZNF385D, ADGRE2, LINC00639, ADGRD1, PRELP, TGFBI, FAM47E -STBD1, TSPAN15, SLC2A1, IER3, AJUBA, NTN4, CEMIP, CX3CL1, SLC16A3, STC2, THBD, IGFBP2, TNFRSF1B, ANXA3, NRP1, CD248, CDC42EP2, IGSF3, ETNPPL, APCDD1, GNAZ, S1PR2, STK26, RDH10, RRAD, TNFAIP2, IL15RA, ITGB4, CD14, CAMK2B, SYNJ2, PIR, FAM156A, TPRA1, F2R, RGL1, GATAD2B, SLC6A6, NEDD4L, M N1, ATF3, ADCY9, SASH1, SLC22A23, SLC7A5, ANXA11, ZNF70, CD58, IL6R, ATXN1, FHL2, GABRA2, CSGALNACT1, S100A4, BIN1, PAPSS2, LMO4, RAB31, EPM2A,A method selected from one or more of the following: TP53INP1, SESN3, LGI2, C14orf132, ENHO, GCSHP5, LRATD1, LGALS7B, MARCHF3, CRYAB, ID1, PLPP4, RPSAP58, EEF1A1P5, TPPP3, MBNL1-AS1, CCDC171, TMEM189-UBE2V1, AFF2, SCIN, ID3, RUBCNL, EFEMP1, MYRIP, MEST, MDFI, LY6D, RHBDL3, SAMD11, OR2S1P, HSPE1-MOB4, LGALS7, HPGD, SULT1E1, LINC00598, or ONECUT2.

[0241] Embodiment 37 A method for treating an animal having a disease or disorder, the method comprising the step of administering to the animal a therapeutically effective amount of a pharmaceutical composition comprising (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol, wherein the animal is selected by alteration genes that respond in cells to treatment with (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol, and the genes are selected from one or more of the following 14 genes: ICA1, MRPS9, NRIP3, NECTIN3, GATD3A, COL5A3, EPHA3, MFF_DT, IGFBP5, ROCK1P1, LYPD6, DKK2, CAP2, or GRM4.

[0242] Embodiment 38 A method for treating an animal having a disease or disorder, the method comprising the step of administering to the animal a therapeutically effective amount of a pharmaceutical composition comprising (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol, wherein the animal is selected by alteration genes that respond in cells to treatment with (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol, and the genes are selected from one or more of the following 14 genes: ICA1, MRPS9, NECTIN3, EPHA3, MFF_DT, ROCK1P1, LYPD6, DKK2, CAP2, or GRM4.

[0243] Embodiment 39 The method according to any one of Embodiments 36 to 38, wherein the cells are derived from a cell type or tissue selected from one or more of the following: adrenal gland, bone marrow, brain, breast, bronchi, caudate nucleus, cerebellum, cerebral cortex, cervix, uterus, colon, endometrium, epididymis, esophagus, fallopian tube, gallbladder, myocardium, hippocampus, kidney, liver, lung, lymph node, nasopharynx, oral mucosa, ovary, pancreas, parathyroid gland, peripheral blood mononuclear cells (PBMCs), placenta, prostate, rectum, salivary gland, seminal vesicle, skeletal muscle, skin, small intestine (including duodenum, jejunum, and ileum), smooth muscle, spleen, stomach, testis, thyroid gland, tonsil, bladder, or vagina.

[0244] Embodiment 40 The disease or disorder is age-related macular degeneration, Alzheimer's disease (AD), amyotrophic lateral sclerosis (ALS), atherosclerosis, autism spectrum disorder (ASD), benign focal muscular atrophy, cerebral infarction, Creutzfeldt-Jakob disease, Crohn's disease, Duchenne palsy, Friedreich's ataxia, frontotemporal dementia (FTD), glaucoma, hereditary spastic paraplegia (HSP), Huntington's disease (HD), inclusion body myositis (IBM), inflammatory bowel disease, ischemia, Kugelberg-Wellander syndrome, Lewy body disease (LBD), multiple sclerosis (MS), myocardial infarction, necrotizing enterocolitis, neurofibromatosis type 1, Paget's disease of bone (PDB), Parkinson's disease (PD), primary lateral sclerosis (PLS), progressive bulbar palsy (PBP), The method according to any one of embodiments 36 to 39, wherein the patient is one or more selected from among muscular atrophy (PMA), pseudobulbar palsy, spinal muscular atrophy (SMA), ulcerative colitis, balocin-containing protein (VCP) related disorders, or Werdnig-Hoffmann disease, transient ischemic attack, ischemia, cerebral hemorrhage, senile cataract, retinal ischemia, retinal vasculitis, Brown-Bearet-van Lehr syndrome, Eels disease, meningitis and encephalitis, post-traumatic stress disorder, Charcot-Marie-Tooth disease, macular degeneration, hereditary Leber's optic atrophy (Kennedy disease), presenile dementia, depressive disorder, temporal lobe epilepsy, hereditary Leber's optic atrophy, cerebrovascular disease, subarachnoid hemorrhage, or schizophrenia.

[0245] Embodiment 41 The method according to any one of Embodiments 36 to 40, wherein the animal is a mammal.

[0246] Embodiment 42 The method according to Embodiment 41, wherein the mammal is a non-human animal.

[0247] Embodiment 43 The method according to Embodiment 41, wherein the mammal is a human.

[0248] Embodiment 44: The method according to any one of Embodiments 36 to 43, wherein (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol is administered in a dose of 0.12 mg / kg or more.

[0249] Embodiment 45 The method according to any one of Embodiments 36 to 43, wherein (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol is administered at a dose of 5 to 5000 mg / day.

[0250] Embodiment 46 The method according to any one of Embodiments 36 to 43, wherein (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol is administered parenterally, enterally, or topically.

[0251] Embodiment 47 The method according to claim 46, wherein 6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol is administered by oral, sublingual, buccal, pulmonary, intranasal, intravenous, intramuscular, or subcutaneous administration.

[0252] Embodiment 48 Use of (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol for the preparation of a medicament for treating a human having the disease according to Embodiment 40.

[0253] Embodiment 49 The method according to one of Embodiments 1 to 21, wherein increasing, decreasing, or changing is determined relative to a corresponding measurement prior to the step of contacting the cell with (6aS)-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-10,11-diol.

[0254] Embodiment 50 The method according to Embodiment 49, wherein the cell is a cell of a human subject, and the human subject has Alzheimer's disease, amyotrophic lateral sclerosis (ALS), Friedreich's ataxia, Huntington's disease (HD), or Parkinson's disease (PD).

[0255] Embodiment 51 A) Predicting the effectiveness of treating a disease or disorder in a subject, wherein the treatment comprises a first administration of an active pharmaceutical agent to a first cell of the subject and optionally a second administration of the active pharmaceutical agent to a second cell of the subject, wherein the first cell has a first cell type, the second cell has the first cell type, and the active pharmaceutical agent has the following formula

Chemical formula

[0256] Embodiment 53 A) Monitoring the treatment of a disease or disorder in a subject, wherein the treatment comprises a first administration of an active pharmaceutical agent to a first cell of the subject and optionally a second administration of the active pharmaceutical agent to a second cell of the subject, wherein the first cell has a first cell type, the second cell has the first cell type, and the active pharmaceutical agent has the following formula

Chemical formula

[0257] Embodiment 53 The method according to Embodiment 51 or 52, wherein the disease or disorder is a neurological disorder.

[0258] Embodiment 54 The method according to Embodiment 51 or 52, wherein the disease or disorder is Alzheimer's disease, amyotrophic lateral sclerosis (ALS), Friedreich's ataxia, Huntington's disease (HD), or Parkinson's disease (PD).

[0259] Embodiment 55 The disease or disorder is age-related macular degeneration, atherosclerosis, autism spectrum disorder (ASD), benign focal muscular atrophy, cerebral infarction, Creutzfeldt-Jakob disease, Crohn's disease, Duchenne palsy, Friedreich's ataxia, frontotemporal dementia (FTD), glaucoma, hereditary spastic paraplegia (HSP), inclusion body myositis (IBM), inflammatory bowel disease, ischemia, Kugelberg-Wellander syndrome, Lewy body disease (LBD), multiple sclerosis (MS), myocardial infarction, necrotizing enterocolitis, neurofibromatosis type 1, Paget's disease of bone (PDB), primary lateral sclerosis (PLS), progressive bulbar palsy (PBP), progressive muscular atrophy (PMA), pseudobulbar palsy, spinal The method according to Embodiment 51 or 52, wherein one or more of the following conditions are selected: muscular atrophy (SMA), ulcerative colitis, balocin-containing protein (VCP) related disorder, or Werdnig-Hoffmann disease, transient ischemic attack, ischemia, cerebral hemorrhage, senile cataract, retinal ischemia, retinal vasculitis, Brown-Bearet-van Lear syndrome, Eels disease, meningitis and encephalitis, post-traumatic stress disorder, Charcot-Marie-Tooth disease, macular degeneration, hereditary Leber's optic atrophy (Kennedy disease), presenile dementia, depressive disorder, temporal lobe epilepsy, hereditary Leber's optic atrophy, cerebrovascular disease, subarachnoid hemorrhage, or schizophrenia.

[0260] embodiment56 1つつるますまするがが、HS6ST2-AS1、CYP1B1、MEGF6、AHRR、CILP、TNS4、CACNA1S、IFNLR1、KSR2、OLFML2B、BCL11B、FGF18、GALNT5、NKAIN1、DAPP1、LINC00599、G PAT3、IL1B、PRAL、FXYD3、CYP26B1、CYP1B1-AS1、CH25H、COLEC12、TIPARP、KIAA1549L、IFI30、NPTX1、ZMIZ1-AS1、ANPEP、ARHGEF16、WFDC11、KIAA1549、 SUSD3、H19、GFRA1、NRG1、NQO1、TCF7、DTX1、PRXL2A、COLGALT2、LUM、HSPB6、SLC16A10、COL24A1、KCND3、SLC1A2、WNT11、CLDN7、RNF128、PTGS1、ZNF385D 、ADGRE2、LINC00639、ADGRD1、PRELP、TGFBI、FAM47E-STBD1、TSPAN15、SLC2A1、IER3、AJUBA、NTN4、CEMIP、CX3CL1、SLC16A3、STC2、THBD、IGFBP2、TNFRSF 1B、ANXA3、NRP1、CD248、CDC42EP2、IGSF3、ETNPPL、APCDD1、GNAZ、S1PR2、STK26、RDH10、RRAD、TNFAIP2、IL15RA、ITGB4、CD14、CAMK2B、SYNJ2、PIR、FAM1 56A、TPRA1、F2R、RGL1、GATAD2B、SLC6A6、NEDD4L、MN1、ATF3、ADCY9、SASH1、SLC22A23、SLC7A5、ANXA11、ZNF70、CD58、IL6R、ATXN1、FHL2、GABRA2、CSGAL NACT1、S100A4、BIN1、PAPSS2、LMO4、RAB31、EPM2A、TP53INP1、SESN3、LGI2、C14orf132、ENHO、GCSHP5、LRATD1、LGALS7B、MARCHF3、CRYAB、ID1、PLPP4、R PSAP58、EEF1A1P5、TPPP3、MBNL1-AS1、CCDC171、TMEM189-UBE2V1、AFF2、SCIN、ID3、RUBCNL、EFEMP1、MYRIP、MEST、MDFI、LY6D、RHBDL3、SAMD11、OR2S1P、The method according to one of embodiments 51 to 55, selected from HSPE1-MOB4, LGALS7, HPGD, SULT1E1, LINC00598, or ONECUT2.

[0261] Embodiment 57 The method according to one of Embodiments 51 to 55, wherein one or more genes are selected from ICA1, MRPS9, NRIP3, NECTIN3, GATD3A, COL5A3, EPHA3, MFF_DT, IGFBP5, ROCK1P1, LYPD6, DKK2, CAP2, or GRM4.

[0262] Embodiment 58 The method according to one of Embodiments 51 to 55, wherein one or more genes are selected from ICA1, MRPS9, NECTIN3, EPHA3, MFF_DT, ROCK1P1, LYPD6, DKK2, CAP2, or GRM4.

[0263] Embodiment 59 The method according to any one of Embodiments 51 to 58, wherein the first differential expression of one or more genes identifies a significantly dysregulated transcript of one or more genes in a first biological sample compared to a second biological sample.

[0264] Embodiment 60: The method according to one of Embodiments 51 to 59, wherein the method is in vitro, ex vivo, or an in vivo method.

[0265] Embodiment 61 The method according to one of Embodiments 51 to 60, wherein the first cell type is derived from a tissue selected from one or more of the following: adrenal gland, bone marrow, brain, breast, bronchi, caudate nucleus, cerebellum, cerebral cortex, cervix, uterus, colon, endometrium, epididymis, esophagus, fallopian tube, gallbladder, myocardium, hippocampus, kidney, liver, lung, lymph node, nasopharynx, oral mucosa, ovary, pancreas, parathyroid gland, peripheral blood mononuclear cells (PBMCs), placenta, prostate, rectum, salivary gland, seminal vesicle, skeletal muscle, skin, small intestine (including duodenum, jejunum, and ileum), smooth muscle, spleen, stomach, testis, thyroid gland, tonsil, bladder, or vagina.

[0266] The method according to any one of Embodiments 51 to 61, wherein the animal is a mammal.

[0267] The method according to Embodiment 62, wherein the mammal is a non-human animal.

[0268] The method according to Embodiment 62, wherein the mammal is a human.

[0269] The method according to any one of Embodiments 51 to 64, wherein the active pharmaceutical agent is administered at a dose of 0.12 mg / kg or more.

[0270] The method according to any one of Embodiments 51 to 65, wherein the active pharmaceutical agent is administered at a dose of 5 to 5000 mg / day.

[0271] The method according to any one of Embodiments 51 to 66, wherein the active pharmaceutical agent is administered parenterally, enterally, or topically.

[0272] The method according to any one of Embodiments 51 to 67, wherein the active pharmaceutical agent is administered by oral, sublingual, buccal, pulmonary, intranasal, intravenous, intramuscular, or subcutaneous administration.

[0273] Embodiment 69 F) monitoring the effectiveness of the treatment based on the second differential expression or the third differential expression; G) continuing the treatment with the second administration; H) after the second administration, collecting a third biological sample from the subject, the third biological sample containing the second cells; I) determining the second differential expression of one or more genes compared to the first biological sample or compared to the third biological sample, further comprising the method according to any one of Embodiments 51 to 68.

[0274] Embodiment 70 The method according to one of Embodiments 51-69, wherein the subject experiences an increase in p62 levels, an increase in LC3 levels, or a decrease in mitochondrial morphological factors, or a combination thereof, as a result of a first dose, a second dose, or both.

[0275] Embodiment 71 The method according to one of Embodiments 51 to 70, wherein the first cell type is a brain cell derived from brain tissue selected from the cerebrum, cerebellum, diencephalon, or brainstem, and optionally the brain cell is an induced brain cell.

[0276] Embodiment 72 The method according to one of Embodiments 51 to 70, wherein the first cell type is a brain cell selected from neurons, astrocytes, oligodendrocytes, or microglia, and optionally the brain cell is an induced brain cell.

[0277] Embodiment 73 The method according to one of Embodiments 51 to 70, wherein the first cell type is a brain cell selected from sensory neurons, motor neurons, interneurons, or cerebral neurons, and optionally the brain cell is an induced brain cell.

[0278] Embodiment 74: The method according to one of Embodiments 51 to 73, wherein the method is a method for treating a disease or disorder.

[0279] Embodiment 75 A method for treating Alzheimer's disease, amyotrophic lateral sclerosis (ALS), Friedreich's ataxia, Huntington's disease (HD), or Parkinson's disease (PD) in a subject requiring the treatment, 1) At a first dose of at least 0.12 mg / kg, and optionally at a dose of 5 to 5000 mg / day, the active pharmaceutical agent is given by the following formula: [ka] Or administering an active pharmaceutical agent having a pharmaceutically acceptable salt thereof to a subject parenterally, enterally, or topically; 2) Measuring the levels of p62, LC3, or mitochondrial morphological factors, or combinations thereof, in a biological sample containing cells, the cells may include brain cells derived from brain tissue selected from the cerebrum, cerebellum, diencephalon, or brainstem, and optionally, the brain cells are induced brain cells, wherein the measurement includes translatome profiling; 3) A method comprising treating Alzheimer's disease, amyotrophic lateral sclerosis (ALS), Huntington's disease (HD), Lou Gehrig's disease, or Parkinson's disease (PD) in a subject with a second dose greater than the first dose, if the subject does not experience an increase in p62 levels, an increase in LC3 levels, or a decrease in mitochondrial morphological factors, or a combination thereof, as a result of administering a first dose.

[0280] Unless otherwise specified, all numerical values ​​used in the specification and claims, representing properties such as the amount of components and molecular weight, reaction conditions, etc., should be understood in all cases to be modified by the term “about.” As used herein, the terms “about” and “approximately” mean within 10–15%, preferably within 5–10%. Thus, unless otherwise specified, the numerical parameters described in the specification and appended claims are approximations that may vary depending on the desired properties to be obtained by the invention. At the very least, and not as an attempt to limit the application of the doctrine of equivalents to the scope of the claims, each numerical parameter should be interpreted by taking into account the number of significant figures reported and by applying common rounding techniques. Although the numerical ranges and parameters describing the broad scope of the invention are approximations, the numerical values ​​described in specific examples are reported as accurately as possible. However, any numerical value inherently contains certain errors that inevitably arise from the standard deviation observed in their respective test measurements.

[0281] In the context describing the invention (particularly in the context of the following claims), the terms “a,” “an,” “the,” and similar referents should be construed to encompass both singular and plural unless otherwise indicated herein or explicitly refuted by the context. The enumeration of ranges of values ​​herein is intended to serve merely as a simplified way of referring individually to each distinct value contained within that range. Unless otherwise indicated herein, each distinct value is incorporated into the specification as if it were individually enumerated herein. All methods described herein may be carried out in any suitable order unless otherwise indicated herein or explicitly refuted by the context. Any examples or use of exemplary language (e.g., “~etc.”) provided herein are intended merely to better illustrate the invention and do not impose any limitations on the scope of the invention as separately claimed. No language in the specification should be construed to indicate any non-claimed element essential to the carrying out of the invention.

[0282] The grouping of alternative elements or embodiments of the invention disclosed herein should not be construed as limitation. Each group member may be referred to and claimed individually or in any combination with other members of that group or other elements found herein. For convenience and / or patentability reasons, it is anticipated that one or more members of a group may be included in or excluded from a group. Where such inclusion or exclusion occurs, this specification shall be deemed to include the modified group and thus satisfy the description requirements of all Markush groups used in the appended claims.

[0283] Specific embodiments of the Invention are described herein, including the best mode known to the inventors for carrying out the Invention. Naturally, variations of these described embodiments will be apparent to those skilled in the art by reading the foregoing description. The inventors expect that those skilled in the art will appropriately adopt such variations, and the inventors intend that the Invention may be carried out in ways other than those specifically described herein. Accordingly, the Invention includes all modifications and equivalents of the subject matter described in the claims appended herein, to the extent permitted by applicable law. Furthermore, unless otherwise indicated herein, or unless it is clearly inconsistent with the context, any combination of the elements described herein in all possible variations thereof is incorporated into the Invention.

[0284] Certain embodiments disclosed herein may be further limited in the claims by using the phrases "consisting of" or "consisting essentially of". Where used in a claim, whether as filed or added by amendment, the transitional phrase "consisting of" excludes any element, process, or component not specified in the claim. The transitional phrase "consisting essentially of" limits the scope of the claim to those that do not substantially affect the specified material or process and the basic and novel features. Embodiments of the invention as so claimed are described and made implementable herein, either implicitly or explicitly.

[0285] Furthermore, numerous references to patents and publications are made throughout this specification. Each of the above-mentioned references and publications is incorporated herein by reference in its entirety.

[0286] In conclusion, it should be understood that the embodiments of the present invention disclosed herein are illustrative of the principles of the present invention. Other modifications that may be adopted are within the scope of the invention. Accordingly, alternative configurations of the present invention may be used in accordance with the teachings herein, not as limitations but as examples. Therefore, the present invention is not limited to those precisely shown and described herein.

Claims

1. A) Predicting the effectiveness of treatment for a disease or disorder in a subject, wherein the treatment comprises a first administration of an active pharmaceutical agent to first cells of the subject, wherein the first cells have a first cell type, and optionally a second administration of the active pharmaceutical agent to second cells of the subject, wherein the second cells have the first cell type, and the active pharmaceutical agent is given by the following formula 【Chemistry 1】 or predict having a pharmaceutically acceptable salt thereof; B) Before the first administration, a first biological sample containing control cells is collected from the subject, wherein the control cells have the first cell type; C) After the first administration and optionally before the second administration, a second biological sample is taken from the subject, wherein the second biological sample contains the first cells; D) Determining the first differential expression of one or more genes in the second biological sample compared with the first biological sample; E) A method comprising predicting the effectiveness of the treatment based on the first differential manifestation.

2. A) Monitoring the treatment of a disease or disorder in a subject, wherein the treatment includes a first administration of an active pharmaceutical agent to a first cell of the subject, wherein the first cell has a first cell type, and optionally a second administration of the active pharmaceutical agent to a second cell of the subject, wherein the second cell has the first cell type, and the active pharmaceutical agent is the following formula 【Chemistry 2】 or a pharmaceutically acceptable salt thereof, to be monitored; B) Before the first administration, a first biological sample containing control cells is collected from the subject, wherein the control cells have the first cell type; C) After the first administration and optionally before the second administration, a second biological sample is taken from the subject, wherein the second biological sample contains the first cells; D) Determining the first differential expression of one or more genes in the second biological sample compared with the first biological sample; E) A method comprising monitoring the treatment based on the first differential expression.

3. The method according to claim 1 or 2, wherein the disease or disorder is a neurological disease.

4. The method according to claim 1 or 2, wherein the disease or disorder is Alzheimer's disease, amyotrophic lateral sclerosis (ALS), Friedreich's ataxia, Huntington's disease (HD), or Parkinson's disease (PD).

5. The aforementioned diseases or disorders include age-related macular degeneration, atherosclerosis, autism spectrum disorder (ASD), benign focal muscular atrophy, cerebral infarction, Creutzfeldt-Jakob disease, Crohn's disease, Duchenne palsy, Friedreich's ataxia, frontotemporal dementia (FTD), glaucoma, hereditary spastic paraplegia (HSP), inclusion body myositis (IBM), inflammatory bowel disease, ischemia, Kugelberg-Wellander syndrome, Lewy body disease (LBD), multiple sclerosis (MS), myocardial infarction, necrotizing enterocolitis, neurofibromatosis type 1, Paget's disease of bone (PDB), primary lateral sclerosis (PLS), progressive bulbar palsy (PBP), progressive muscular atrophy (PMA), pseudobulbar palsy, The method according to claim 1 or 2, wherein one or more of the following conditions are selected: spinal muscular atrophy (SMA), ulcerative colitis, balocin-containing protein (VCP) related disorder, Werdnig-Hoffmann disease, transient ischemic attack, ischemia, cerebral hemorrhage, senile cataract, retinal ischemia, retinal vasculitis, Brown-Bearet-van Lear syndrome, Eels disease, meningitis and encephalitis, post-traumatic stress disorder, Charcot-Marie-Tooth disease, macular degeneration, hereditary Leber's optic atrophy (Kennedy disease), presenile dementia, depressive disorder, temporal lobe epilepsy, hereditary Leber's optic atrophy, cerebrovascular disease, subarachnoid hemorrhage, or schizophrenia.

6. The aforementioned 1つなれますいまするがが、HS6ST2-BS、CEP1B1、MEG5620AA、-LP、TNS4、CACNA1S、IFNLA1、KSR2、-LFML2B、BLL11B、FGF18、LLNT5、NKAIN1、DAPP1、LINC00599 、GPAT3、-L1B、PRAL、FEYD3、CYP26B1、CYPB1-AS1、CH25H、COLEC12、TIPAR 、KIAA1549L、DIC0、KEE1、ZMIZ-AS1、ANPEP、ARGE55、WFDC11、KIAA1549 、USD3、H19、GFRA1、NRG1、NQO1、TCF7、DTX1、PR^L2A、COLGALD2、L@M、HSPB6 、SLC16A10、COL24A1、KCID3、SLC1A2、WNT11、LDE7、RNF128、PTGS1、ZNF385 D、ADGRE2、LINC00639、ADGRD1、PREL@、EGFNE-STBD1-DGP11DLC 2A1、-ER3、ANUBA、EEE2、CEMIP、CE3CL1、SLC16A3、STC2、THHBD、EGFBP2、EFI F1B、ANXA3、NRP1、CD248、CDC42EP2、GGFK3、ETNPPL、APCDD1、G1Z、S1PRR2、S TK26、RDX10RRAD、TDFAIP2、IL11RA、>GB4、CD14、CAMK2B、SYEAD、PIR、1AM 156A、DDRA1、D2R、RGL1、GADAD2B、LC6A5、NEDD4L、MN1、DK3、AO9、4II 、QLC22A23、QLC7A5、ANXA11、ZNF70、CD58、IL6R、AEN1、FHL2、GABRAAGGGAL NACT1、S100A4、BIN1、PAPSS2、LMO4、RAB31、EPMM2A、TP53.NP1、SEEN3、LGI2、 C14orf132、ENHO、GCHHP5、LRATD1、LGALS7B、MARCHF3、CRYAB、ID1、PLPP4、R ΠBA@8、EE1A1P5、9PPPP3、MBNL1-AS1、ACDC171、TMEM189-2BE271-11125 IN、ID3、RUBCNL、EFEMP1、MYRIP、MEST、MDKI、LY6D、RHBDL3、AMD11、OR2@1The method according to one of claims 1 to 5, selected from HSPE1-MOB4, LGALS7, HPGD, SULT1E1, LINC00598, or ONECUT2.

7. The method according to one of claims 1 to 5, wherein one or more of the genes are selected from ICA1, MRPS9, NRIP3, NECTIN3, GATD3A, COL5A3, EPHA3, MFF_DT, IGFBP5, ROCK1P1, LYPD6, DKK2, CAP2, or GRM4.

8. The method according to one of claims 1 to 5, wherein one or more of the genes are selected from ICA1, MRPS9, NECTIN3, EPHA3, MFF_DT, ROCK1P1, LYPD6, DKK2, CAP2, or GRM4.

9. The method according to one of claims 1 to 8, wherein the first differential expression of the one or more genes identifies a significantly dysregulated transcript of the one or more genes in the first biological sample compared with the second biological sample.

10. The method according to one of claims 1 to 9, which is an in vitro or ex vivo method.

11. The method according to one of claims 1 to 10, wherein the first cell type is derived from a tissue selected from one or more of the following: adrenal gland, bone marrow, brain, breast, bronchus, caudate nucleus, cerebellum, cerebral cortex, cervix, uterus, colon, endometrium, epididymis, esophagus, fallopian tube, gallbladder, myocardium, hippocampus, kidney, liver, lung, lymph node, nasopharynx, oral mucosa, ovary, pancreas, parathyroid gland, peripheral blood mononuclear cells (PBMCs), placenta, prostate, rectum, salivary gland, seminal vesicle, skeletal muscle, skin, small intestine (including duodenum, jejunum, and ileum), smooth muscle, spleen, stomach, testis, thyroid gland, tonsil, bladder, or vagina.

12. The method according to any one of claims 1 to 11, wherein the animal is a mammal.

13. The method according to claim 12, wherein the mammal is a non-human animal.

14. The method according to claim 12, wherein the mammal is a human.

15. The method according to one of claims 1 to 14, wherein the active pharmaceutical agent is administered in a dose of 0.12 mg / kg or more.

16. The method according to one of claims 1 to 15, wherein the active pharmaceutical agent is administered in a dose of 5 to 5000 mg / day.

17. The method according to one of claims 1 to 16, wherein the active pharmaceutical agent is administered parenterally, enterally, or topically.

18. The method according to one of claims 1 to 17, wherein the active pharmaceutical agent is administered orally, sublingually, buccally, in the lung, intranasally, intravenously, intramuscularly, or subcutaneously.

19. F) Monitoring the effectiveness of the treatment based on a second differential expression or a third differential expression; G) Continuing the treatment with the second administration; H) After the second administration, a third biological sample is collected from the subject, wherein the third biological sample contains the second cells. I) The method according to one of claims 1 to 18, further comprising determining the second differential expression of one or more genes in comparison with the first biological sample or in comparison with the third biological sample.

20. The method according to one of embodiments 1 to 19, wherein the subject experiences an increase in p62 levels, an increase in LC3 levels, or a decrease in mitochondrial morphological factors, or a combination thereof, as a result of the first administration, the second administration, or both.

21. The method according to one of claims 1 to 20, wherein the first cell type is a brain cell derived from brain tissue selected from the cerebrum, cerebellum, diencephalon, or brainstem, and optionally the brain cell is an induced brain cell.

22. The method according to one of claims 1 to 20, wherein the first cell type is a brain cell selected from neurons, astrocytes, oligodendrocytes, or microglia, and optionally the brain cell is an induced brain cell.

23. The method according to one of claims 1 to 20, wherein the first cell type is a brain cell selected from sensory neurons, motor neurons, interneurons, or cerebral neurons, and optionally the brain cell is an induced brain cell.

24. A method for treating the aforementioned disease or disorder, according to one of claims 1 to 23.

25. A method for treating Alzheimer's disease, amyotrophic lateral sclerosis (ALS), Friedreich's ataxia, Huntington's disease (HD), or Parkinson's disease (PD) in a person requiring such treatment, wherein the method is 1) At a first dose of at least 0.12 mg / kg, and optionally at a dose of 5 to 5000 mg / day, the following formula 【Transformation 3】 Administering an active pharmaceutical agent having a pharmaceutically acceptable salt thereof to the subject parenterally, enterally, or topically; 2) Measuring the levels of p62, LC3, or mitochondrial morphological factors, or combinations thereof, in a biological sample containing cells, wherein the cells may include brain cells derived from brain tissue selected from the cerebrum, cerebellum, diencephalon, or brainstem, and optionally, the brain cells are induced brain cells, wherein the measurement includes translatome profiling; 3) A method comprising treating Alzheimer's disease, amyotrophic lateral sclerosis (ALS), Friedreich's ataxia, Huntington's disease (HD), or Parkinson's disease (PD) in the subject with a second dose greater than the first dose, if the subject does not experience an increase in p62 levels, an increase in LC3 levels, or a decrease in mitochondrial morphological factors, or a combination thereof, as a result of administering the first dose.

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