GB2601519A - Method of support of survival and protection of neurons by RNA m6A methyltransferase complex METTL3/METTL14 activators - Google Patents

Abstract

A method of support of the dopaminer neuron survival and protecting dopamine neurons from neurotoxin-induced death using compounds that modulate the RNA methylation by activating the METTL3/METTL14 methyltransferase complex wherein said compound is of formula (I), (II), (IV), (V), (VI), (VII) or (VIII) or a pharmaceutically acceptable salt thereof: (I) (II) (III) (IV) (V) (VI) (VII) (VIII) wherein R1 and R2 are independently selected from the group consisting of H, alkyl, aryl, aralkyl, acyl, alkoxycarbonyl, aryloxycarbonyl, aralkoxycarbonyl, carbamoyl, alkylcarbamoyl, and dialkylcarbamoyl, aminoalkyl, aminoalaryl. These compounds are useful in the treatment of Parkinson's disease, amyotrophic lateral sclerosis, Alzheimer’s disease, drug addiction, traumatic brain injury, psychosis, depression, multiple sclerosis and nausea. These compounds support the survival of dopamine neurons and protect them from cell death and apoptosis. The compounds belong to the class that modulate RNA methylation by activation of the RNA m6A methyltransferase complex METTL3/METTL14.

Description

METHOD OF SUPPORT OF SURVIVAL AND PROTECTION OF NEURONS BY RNA m6A METHYLTRANSFERASE COMPLEX METTL3/METTL14 ACTIVATORS Inventors: Mart Saa17770; Li-Ying Yu, Neinar Sell, Si177011a Se/berg, Mali Kareholt Assignee: Chentestrned, Ltd

TECHNICAL FIELD

The presently disclosed subject matter generally relates to the epitrancriptomic regulation of ribonucleic acid (RNA) methylation through small-molecule activators of the RNA m6A methyltransferase complex METTL3/NIETTL14AVTAP and their effects on the neuronal cell survival and neuroprotection.

BACKGROUND ART

Chemical modifications of RNA have been shown to have critical impact on many cellular functions, such as proliferation, survival and differentiation, mostly through regulation of RNA stability (Tzelepis, et al, 2019; Anreiter, et al, 2020). The most abundant modification in eukaryotic messenger RNA is N6-methyladenosine (m6A) that affects its splicing, intracellular distribution, translation, intracellular transport and cytopl asmi c degradation, playing thus a crucial role in regulating cell differentiation, neuronal signalling, carcinogenesis and immune tolerance Nally et al., 2016; Roundtree et al., 2017). The m6A methylation level in RNA is regulated by specific enzymes, i.e. the RNA methyltransferases, RNA methylases and RNA reader proteins. Those include the RNA methyltransferase enzyme complex METTL3/METTL14/WTAP consisting of three components: METTL3 (methyltransferase-like 3), METTL14 (methyltransferase-like 14), and WTAP (Wilm's tumour-1 -associated protein), called also the "writer" enzyme (Liu et al, 2014; Meyer et al, 2017); the methyltransferase-like protein 16 (METTL16) (Brown, et al, 2016); the RNA demethylases FTO (fat mass and obesity-associated protein)(Jia et at, 2011; Zheng et al., 2013) and AlkBH5 (AlkB family member 5) (Wang, et al, 2020), called "erasers". The fate of the RNA in post-transcriptomic processes is also directed by the "reader" enzymes that recognize specific m6A methylation in RNA. Several RNA reader proteins and enzymes have been identified (Meyer et al., 2017; Patil et al., 2017), including YTHDF1 (YTH N6-Methyladenosine RNA Binding Protein 1), YTHDF2 (YTH N6-Methyladenosine RNA Binding Protein 2) YTHDF3 (YTH N6-Methyladenosine RNA Binding Protein 3), YTHDC1 (YTH domain-containing protein 1) and YTHDC2 (YTH domain-containing protein 2) (Wojtas et al., 2017). These three types of enzymes collectively coordinate the m6A RNA methylome in the eukar)..rotic cell.

As mentioned above, the RNA m6A modification can be important in brain development, neuron signalling and neuronal disorders (Angelova, et al, 2018; Jung, et al, 2018; Noack, et al, 2019; Widagdo, et al, 2018; Du, et al, 2019; Livneh, et al, 2020). For instance, it has been demonstrated that m6A-dependent mRNA decay is critical for proper transcriptional prepatterning in mammalian cortical neurogenesis (Yoon, et al, 2017). Weng et al, had uncovered an epitranscriptomic mechanism wherein axonal injury elevates m6A levels and signalling to promote protein translation, including regeneration-associated genes, which is essential for functional axon regeneration of peripheral sensory neurons (Weng, et al, 2018).

It has been shown on the limited patient group (Chinese Han population) that m6A genes may play a role in conferring risk of dementia (Du, et al, 2015). Different m6A regulating enzymes are involved directly in the processes of learning and memory. Through its binding protein YTHDF1, m6A promotes protein translation of target transcripts in response to neuronal stimuli in the adult mouse hippocampus, thereby facilitating learning and memory (Shi, et al, 2018). Similarly, the RNA m6A demethylase FTO is expressed in adult neural stem cells and neurons and displays dynamic expression during postnatal neurodevelopment. It has been demonstrated that the deficiency of FTO could reduce the proliferation and neuronal differentiation of adult neural stem cells in vivo, leading to impaired learning and memory (Li, et al, 2017). In addition, the inactivation of the Fto gene, encoding a nucleic acid demethylase, impairs dopamine receptor type 2 (D2R) and type 3 (D3R)-dependent control of neuronal activity and behavioural responses (Hess, et al, 2013). The m6A homeostasis in neurons are controlled by more than one regulator. Thus, it was found that global m6A modification of mRNAs is down-regulated in 6-0HDA-induced rat pheochromocytoma PC12 cells originating from adrenal medulla and the striatum of 6-0HDA treated rat brain that is the animal model of Parkinson's disease (PD).. The reduction of the m6A level in PC12 cells by overexpressing a nucleic acid demethylase, FTO, or by m6A inhibitor cycloleucine could induce the expression of N-methyl-D-aspartate (NMIDA) receptor 1, and elevate oxidative stress and Ca2+ influx, resulting in PC12 cell apoptosis (Chen, et al, 2019). Based on above, the dysregulation of the m6A can possibly be related to neurodegeneration in Parkinson's disease. In summary, either the compensatory upregulation or downregulation of m6A could be needed in neuronal cells, depending on their physiological or pathological state.

Recently, some of us have demonstrated that the eukaryotic mRNA methylation at 6-methyladenosine position can be enhanced by small molecule activator compounds (Selberg, et al., 2018; Selberg, et al, 2019). As referred above (Chen, et al, 2019), the downregulation of m6A is involved in dopamine neuron death. Therefore, the m6A activator compounds can support the homeostasis of m6A in dopamine neurons, support their survival in stress conditions and protect them from neurotoxin-induced cell death.

SUMMARY OF INVENTION

The present invention is related to a method of supporting the survival of the neuronal cells and protecting them from cell death induced by neurotoxins or injury using the RNA m6A methyltransferase complex METTL3/METTL14/WTAP activators. Also disclosed are the compounds, or salts or esters thereof, which can protect and support the survival of the neuronal cells. The compounds described can be used for treating Parkinson's disease, amyotrophic lateral sclerosis, Alzheimer's disease, drug addiction, traumatic brain injury, psychosis, depression, multiple sclerosis and nausea by administering to the subject one or more compounds, or a pharmaceutically acceptable salt thereof The "summary of invention" heading is not intended to be restrictive or limiting. The invention also includes all aspects described in the detailed description or figures as originally filed. The original claims appended hereto also define aspects that are contemplated as the invention and are incorporated into this summary by reference.

In addition to the foregoing, the invention includes, as an additional aspect, all embodiments of the invention narrower in scope in any way than the variations specifically mentioned above. For example, although aspects of the invention may have been described by reference to a genus or a range of values for brevity, it should be understood that each member of the genus and each value or sub-range within the range is intended as an aspect of the invention.

Likewise, various aspects and features of the invention can be combined, creating additional aspects which are intended to be within the scope of the invention. Although the applicant(s) invented the full scope of the claims appended hereto, the claims appended hereto are not intended to encompass within their scope the prior art work of others. Therefore, in the event that statutory prior art within the scope of a claim is brought to the attention of the applicants by a Patent Office or other entity or individual, the applicant(s) reserve the right to exercise amendment rights under applicable patent laws to redefine the subject matter of such a claim to specifically exclude such statutory prior art or obvious variations of statutory prior art from the scope of such a claim Variations of the invention defined by such amended claims also are intended as aspects of the invention

BRIEF DESCRIPTION OF DRAWINGS

Figure I. The effect of the compounds (IV) -(VII) on the survival of the dopamine neurons.

Figure 2. The effect of the compound (VIII) on the survival of the dopamine neurons.

Figure 3. Amphetamine-induced ipsilateral rotations at 2 weeks (pre-treatment), at 6 weeks and 9 weeks postlesion.

DETAILED DESCRIPTION OF INVENTION

Disclosed herein are compounds and methods of supporting the survival of the neuronal cells and protecting them from neurotoxin-or injury-induced cell death by using the RNA m6A methyltransferase complex METTL3/METTL14/WTAP activators. In some variations of the invention, the compound is administered in a composition that also includes one or more pharmaceutically acceptable diluents, adjuvants, or carriers The compound can be a small molecule. In some embodiments, the compound supporting the survival of neuronal cells has a structure of Formula (I), ft (I) wherein: RI and R2 are independently selected from the group consisting of H, alkyl, aryl, aralk yl, acyl, al k oxycarbonyl, aryl oxvcarbonyl, aralkoxycarbonyl, carb am oyl, alkylcarbamoyl, and dialkylcarbamoyl, aminoalkyl, aminoalaryl; or a pharmaceutically acceptable salt thereof In some embodiments, the compound supporting the survival of neuronal cells has a structure of Formula (II) HN'e-----''''' Icy r,H

RI

wherein R1 and R2 are independently selected from the group consisting of H, alkyl, aryl, alkylenearyl, acyl, alkoxycarbonyl, aryl oxycarbonyl, alkylenearyloxycarbonyl, carbamoyl, alkylcarbamoyl, dialkylcarbamoyl, and alkyleneamino or a pharmaceutically acceptable salt thereof In some embodiments, the compound supporting the survival of neuronal cells has a structure of Formula (III) wherein RI and R2 are independently selected from the group consisting of H, alkyl, aryl, alkylenearyl, acyl, alkoxycarbonyl, aryloxycarbonyl, alkylenearyloxycarbonyl, carbamoyl, alkylcarbamoyl, dialkylcarbamoyl, and alkyleneamino or a pharmaceutically acceptable salt thereof In some embodiments, the compound supporting the survival of neuronal cells has a structure 15 of Formula (IV) (IV) or a pharmaceutically acceptable salt thereof In some embodiments, the compound supporting the survival of neuronal cells has a structure of Formula (V) (V) or a pharmaceutically acceptable salt thereof In some embodiments, the compound supporting the survival of neuronal cells has a structure of Formula (VI) (VI) or a pharmaceutically acceptable salt thereof In some embodiments, the compound supporting the survival of neuronal cells has a structure of Formula (VII) (VII) or a pharmaceutically acceptable salt thereof In some embodiments, the compound supporting the survival of neuronal cells has a structure of Formula (VIII) (VIII) or a pharmaceutically acceptable salt thereof The compounds described can be used for treating Parkinson's disease, amyotrophic lateral sclerosis, Alzheimer's disease, drug addiction, traumatic brain injury, psychosis, depression, multiple sclerosis and nausea by administering to the subject one or more compounds, or a pharmaceutically acceptable salt thereof As used herein, the term "alkyl" refers to straight chained and branched hydrocarbon groups containing carbon atoms, typically methyl, ethyl, and straight chain and branched propyl and butyl groups. Unless otherwise indicated, the hydrocarbon group can contain up to 20 carbon atoms. The term "alkyl" includes "bridged alkyl," i.e., a C.sub.6-C.sub.16 bicyclic or polycyclic hydrocarbon group, for example, norbornyl, adamantyl, bicyclo[2.2.2]octyl, bicyclo[2.2. Hheptyl, bicyclop.2. Hoctyl, or decahydronaphthyl. Alkyl groups optionally can be substituted, for example, with hydroxy (OH), halo, amino, and sulfonyl. An "alkoxy" group is an alkyl group having an oxygen substituent, e.g., -0-alkyl.

The term "alkenyl" refers to straight chained and branched hydrocarbon groups containing carbon atoms having at least one carbon-carbon double bond. Unless otherwise indicated, the hydrocarbon group can contain up to 20 carbon atoms. Alkenyl groups can optionally be substituted, for example, with hydroxy (OH), halo, amino, and sulfonyl.

As used herein, the term "alkylene" refers to an alkyl group having a further defined substituent. For example, the term "alkylenearyl" refers to an alkyl group substituted with an aryl group, and "alkyleneamino" refers to an alkyl groups substituted with an amino group. The amino group of the alkyleneamino can be further substituted with, e.g., an alkyl group, an alkylenearyl group, an ai-y1 group, or combinations thereof The term "alkenylene" refers to an alkenyl group having a further defined substituent As used herein, the term "aryl" refers to a monocyclic or polycyclic aromatic group, preferably a monocyclic or bicyclic aromatic group, e.g., phenyl or naphthyl. Unless otherwise indicated, an aryl group can be unsubstituted or substituted with one or more, and in particular one to four groups independently selected from, for example, halo, alkyl, alkenyl, OCF.sub.3, NO.sub.2, CN, NC, OH, alkoxy, amino, CO.sub.2H, CO.sub.2alkyl, aryl, and heteroaryl. Exemplary aryl groups include, but are not limited to, phenyl, naphthyl, tetrahydronaphthyl, chlorophenyl, methylphenyl, methoxyphenyl, trifluoromethylphenyl, nitrophenyl, 2,4-methoxychlorophenyl, and the like. An "aryloxy" group is an aryl group having an oxygen substituent, e.g., --0-aryl.

As used herein, the term "acyl" refers to a carbonyl group, e.g., C(0). The acyl group is further substituted with, for example, hydrogen, an alkyl, an alkenyl, an aryl, an alkenylaryl, an alkoxy, or an amino group. Specific examples of acyl groups include, but are not limited to, al koxycarb onyl (e.g., C(0)--Oal kyl); aryl oxycarbonyl (e.g., C(0)--Oary1); alkylenearyloxycarbonyl (e.g., C(0)--Oalkyleneary1); carbamoyl (e.g., C(0)--NH.sub.2); alkylcarbamoyl (e.g., C(0)--NH(alkyl)) or dialkylcarbamoyl (e.g., C(0)--NH(alkyl).sub.2).

As used herein, the term "amino" refers to a nitrogen containing substituent, which can have zero, one, or two alkyl, alkenyl, aryl, alkylenearyl, or acyl substituents. An amino group having zero substituents is --NH.sub.2.

As used herein, the term "halo" or "halogen" refers to fluoride, bromide, iodide, or chloride As used herein, the term "pharmaceutically acceptable salt" refers to those salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response and the like, and are commensurate with a reasonable benefit/risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, S. M. Berge, et al. describes pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 66: 1-19 (1977). The salts can be prepared in situ during the final isolation and purification of the compounds of the invention, or separately by reacting the free base function with a suitable organic acid or inorganic acid.

Examples of pharmaceutically acceptable nontoxic acid addition salts include, but are not limited to, salts of an amino group formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid or with organic acids such as acetic acid, maleic acid, tartaric acid, citric acid, succinic acid lactobionic acid or malonic acid or by using other methods used in the art such as ion exchange Other pharmaceutically acceptable salts include, but are not limited to, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and the like. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Further pharmaceutically acceptable salts include, when appropriate, nontoxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, alkyl having from 1 to 6 carbon atoms, sulfonate and aryl sulfonate.

EXAMPLES

The following Examples have been included to provide illustrations of the presently disclosed subject matter. In light of the present disclosure and the general level of skill in the art, those of skill will appreciate that the following Examples are intended to be exemplary only and that numerous changes, modifications and alterations can be employed without departing from the spirit and scope of the presently disclosed subject matter.

Example I. Primary cultures of midbrain dopamine neurons and m6A regulator treatment.

The midbrain floors were dissected from the ventral mesencephalic of 13 days old NMRI strain mouse embryos. The tissues were incubated with 0.5% trypsin (103139, MP Biomedical) in HBSS (Ca2+/Mg2+-free) (14170112, Invitrogen) for 20 min at 37 C, then mechanically dissociated. Cells were plated onto the 96-well plates coated with poly-Lornithine (Sigma-Aldrich). Equal volumes of cell suspension were plated onto the center of the dish. The cells were grown for 5 days with different concentrations of m6A regulators.

Human recombinant GDNF (100 ng/ml) (Icosagen) or a medium without any neurotrophic compound added were used as positive and negative controls, respectively.

After growing 5 days, the neuronal cultures were fixed and stained with anti-Tyrosine Hydroxylase antibody (MAB318, Millipore Bioscience Research Reagents. Images were acquired by CellInsight high-content imaging equipment. lmmunopositive neurons were counted by CellProfiler software and the data was analysed by Cell Profiler analyst software.

(Yu, et al, 2008; Mahato et al, 2020). The results are expressed as % of cell survival compared to GDNF-maintained neurons (Figure 1 and 2). It can be seen that the compounds have some effect already at the 10 nM concentration and comparable to the neurotrophic factor GDNF effect at their 1 000 nM concentration. Therefore, these compounds are efficiently supporting the survival of neurons at growth factor deprivation-induced stress conditions.

Example 2. Neurorestorative effect of m6A mRNA methylation activator (VII) in rat 6-OHDA model of Parkinson's disease.

A chronic 4-week intrastriatal infusion of mRNA methylation activator (VII) was used to examine its effect in 6-0HDA model of PD. Small doses of 6-01-IDA (3x2 Mg) were injected to three different sites in the striatum. Two weeks later pre-rotation tests were done. Rats rotating more than 50 rotations / 2 h were taken to the experiment and divided based on their rotations equally into 4 groups. Targeted animal number was 12 / group. Thereafter brain infusion cannulas connected to Alzet minipumps were implanted into the striatum to infuse compound (VII) for 4-weeks. The neuroprotective and restorative effects were evaluated by d-amphetamine induced rotations (amphetamine sulfate 2.5mg/kg; ip) and cylinder tests.

Treatment groups: male Wistar rats PBS treated control n=12-14 * GDNF dose 3.0 micrograms / 24 h for 4 weeks n=12-14 compound (VII) dose 1, 0.3 micrograms / 24 h for 4 weeks n=12-14 * compound (VII) dose 2, 1.5 micrograms / 24 h for 4 weeks n=12-I4 The results on amphetamine-induced rotations (d-amphetamine 2.5 mg/kg i.p., 120 min) are given in Figure 3. A large decline (about 50-60%) in the ipsilateral rotations can be observed at 6 weeks postlesion for compound (VII), witnessing a major neurorestorative effect in vivo.

At 9 eweeks postlesion also GDNF shows clear effect and compound (VII) at 1.5 micrograms / 24 h very efficiently reduces the lesion size induced by 6-0HDA neurotoxin. Compound (VII) is able to protect dopamine neurons from 6-OHBA lesion, reducing the lesion size and I1 improving the motor behavior of the lesioned animals. The results also show that the similarly to GDNF compound (VII) has long lasting effects, but compound (VII) acts faster than GDNF.

REFERENCES CITED

Angelova MT, Dimitrova D.G, Dinges N, Lence T, Worpenberg L, Carre C, Roignant J.Y. (2018). The Emerging Field of Epitranscriptomics in Neurodevelopmental and Neuronal Disorders. Front. Bioeng. Biotechnol. 6, 46 Anreiter, I., Mir, Q., Simpson, J. T., Janga, S. C., & Soller, M. (2020). New Twists in Detecting mRNA Modification Dynamics. Trends Biotechnol., S0167-7799(20)30166-9.

Brown, J.A., Kinzig, CO., DeGregorio, S.J., Steitz, J.A. (2016). Nlethyltransferase-like protein 16 binds the 3'-terminal triple helix of MALAT1 long noncoding RNA. Proc. Natl. Acad, Sci, U.S.A., H3, 14013-14018.

Chen X, Yu C, Guo M, Zheng X, Ali S, Huang H, Zhang L, Wang S. Huang Y, Qie S. Wang J. (2019) Down-Regulation of m6A mRNA Methylation Is Involved in Dopaminergic 15 Neuronal Death. ACS Chem Neurosci. 10, 2355-2363.

Du, T.; Rao, S.; Wu, L.; Ye, N.; Liu, Z.; Hu, H.; Xiu, J.; Shen, Y.; Xu, Q. (2015) An association study of the m6A genes with major depressive disorder in Chinese Han population. J Affect Di sord. 183, 279-86, Du, K; Zhang, L; Lee, T; Sun, T. (2019). m6A RNA Methylation Controls Neural Development and Is Involved in Human Diseases. Mol, Neurobiol. 56, 1596-1606.

Hess, NI. E., Hess, S., Meyer, K. D., Verhagen, L. A. W., Koch, L., Bronneke, H. S., Dietrich, M. 0., Jordan, S. D., Saletore, Y., Elemento, 0., Belgardt, B. F., Franz, T., Horvath, T. L., Ruther, U., Jaffrey, S. R., Kloppenburg, P., and Bruning, J. C. (2013) The fat mass and obesity associated gene (Fto) regulates activity of the dopaminergic midbrain circuitry. Nat.

Neurosci, 16, 1042-1048.

Jia, G., Fu, Y., Zhao, X., Dai, Q., Zheng, G., Yang, Y., et al. (2011). N6-methyladenos ne in nuclear RNA is a major substrate of the obesity-associated FTO. Nat, Chem, Biol. 7, 885887.

Jung Y.; Goldman D. (2018). Role of RNA modifications in brain and behavior. Genes Brain Behav. 17, e12444 Li, L., Zang, L., Zhang, F., Chen, J., Shen, H., Shu, L., Liang, F., Feng, C., Chen, D., Tao, H., Xu, T., Li, Z., Kang, Y., Wu, H., Tang, L., Zhang, P., Sin, P., Shu, Q., and Li, X. (2017) Fat mass and obesity associated (FTO) protein regulates adult neurogenesis. Hum. Mol, Genet, 26, 2398-2411 Liu J.; Yue Y.; Han D.; Wang X.; Fu Y.; Zhang L., Jia G.; Yu NI., Lu Z., Deng X., Dai Q.; Chen W.; He C. (2014) A METTL3-NIETTL14 complex mediates mammalian nuclear RNA N6-adenosine methylation. Nat. Chem. Biol, 10, 93-95.

Livneh, I.; Moshitch-Moshkovitz, S.; Amariglio, N. et al. (2020). The m6A ep transcriptome: transcriptome plasticity in brain development and function. Nat. Rev. Neuracci. 21, 36-51.

Mahato AK, Kopra J, Renko JM, Visnapuu T, Korhonen I, Pulkkinen N, Bespalov MM, Domanskyi A, Ronken E, Piepponen TP, Voutilainen MH, Tuominen RK, Karelson M, Sidorova YA, Saarma M. (2020) Glial cell line-derived neurotrophic factor receptor Rearranged during transfection agonist supports dopamine neurons in vitro and enhances dopamine release in vivo. ilinv Disord. 35, 245-255.

Maity, A.; Das, B. (2016). N6-methyladenosine Modification in mRNA: Machinery, Function and Implications for Health and Diseases. FEBS Journal. 283, 1607-1630.

Meyer, K.D.; Jaffrey,S.R. (2017). Rethinking m6A readers, writers, and erasers. Alum. Rev.

Cell Dev. Biol. 33, 319-342.

Noack F.; Calegari F. (2018). Epitranscriptomics: A New Regulatory Mechanism of Brain Development and Function. Front. Neurosci. 12, 85.

Patil, D. P.; Pickering, B. F.; and Jaffrey, S. R. (2017). Reading m6A in the transcr ptome: m6A-binding proteins. Trends Cell Biol, 28, 113-127.

Roundtree, IA., Evans, ME., Pan, T. & He, C (2017). Dynamic RNA Modifications in Gene Expression Regulation. Cell 169, 1188-1200.

Selberg, S., Blokhina, D., Kankuri, E., Karelson, NI. (2018). Method of activating RNA methyltransferase complex METTL3/METTL14/WTAP, Pat. application

H

Selberg, S.; Blokhina, D.; Aatonen, M.; Siltanen,A; Mervaala, E.; Kankuri, E.; Karelson, M. (2019) Discovery of small molecules that activate RNA-methylation through cooperative binding to the METTL3/14/WTAP complex active site, Cell Rep. 26, 3762-3771.

Shi, H., Zhang, X., Weng, Y. L., Lu, Z., Liu, Y., Li, J., Hao, P, Zhang, Y., Zhang, F., Wu, Y., Delgado, J. Y., Su, Y., Patel, M. J., Cao, X., Shen, B., Huang, X., Ming, G. L., Zhuang, X., Song, H., He, C., and Zhou, T. (2018) m(6)A facilitates hippocampus-dependent learning and memory through YTHDFI. Nature 563, 249-253.

Tzelepis, K., Rausch, 0. & Kouzarides, T. (2019). RNA-modifying enzymes and their function in a chromatin context. Nat. Struct. Mol. Biol. 26, 858-862.

Wang, J.; Wang, J.; Gu, Q.; et al., (2020). The biological function of m6A demethylase ALKBH5 and its role in human disease, Cancer Cell Int. 20, 347.

Weng, Y. L.; Wang, X.; An, R.; Cassin, J.; Vissers, C.; Liu, Y.; Liu, Y., Xu, T.; Wang, X.; Wong, S.; Joseph, J.; Dore, L. C.; Dong, Q.; Zheng, W.; Jin, P.; Wu, H.; Shen, B.; Zhuang, X.; He, C.; Liu, K.; Song, H.; Ailing, G. L. (2018). Epitranscriptomic m6A Regulation of Axon Regeneration in the Adult Mammalian Nervous System. Neuron, 97(2), 313-325.e6.

Widagdo J.; Anggono V. (2018). The m6A-epitranscriptomic signature in neurobiology: from neurodevelopment to brain plasticity. J. Neurochem, 147, 137-152.

Wojtas, MN.; Pandey, R.R.; Mendel, M.; Homolka, D.; Sachidanandam, R.; Pillar, R.S. (2017). Regulation of m6A Transcripts by the 3'->5' RNA Helicase YTHDC2 Is Essential for a Successful Meiotic Program in the Mammalian Germline, Mol Cell, 68, 374-387.

Yoon, Kt; Ringeling, F.R.; Vissers, C.; Jacob, F.; Pokrass, M.; Jimenez-Cyrus, D.; Su, Y.; Kim, N. S.; Zhu, Y.; Zheng, L.; Kim, S.; Wang, X.; Dore, L.C.; Jin, P.; Regot, S.; Zhuang, X.; Canzar, S.; He, C.; Ming, G.L.; Song, H. (2017). Temporal Control of Mammalian Cortical Neurogenesis by m6A Tvlethylation. Cell. 171, 877-889.e17.

Yu, L.-Y., Saarma, M. and Arumae, U. (2008) Death receptors and caspases but not mitochondria are activated in the GDNF-or BDNF-deprived dopaminergic neurons. J. Neurosci, 28 (30), 7467-7475 Zheng, G. ; Dahl, J. A. ; Niu, Y. ; Fedorcsak, P. ; Huang, C. M. ; Li, C. J. ; Vagbo, C. B. ; Shi, Y. ; Wang, W. L. ; Song, S. H. ; et a1. (2013). ALKBH5 is a mammalian RNA demethylase that impacts RNA metabolism and mouse fertility. Mol Cell, 49, 18-29.

Claims (1)

1. NClaims I. A method of support of the dopaminer neuron survival and protecting dopamine enruosn from neurotoxin-induced death using a compounds that modulate the RNA methylation by activating the METTL3/METTL14 methyltransferase complex, wherein the compound is a small molecule having a structure of Formula (1) (I) wherein: R1 and R2 are independently selected from the group consisting of H, alkyl, aryl, aralkyl, acyl, alkoxycarbonyl, aryloxycarbonyl, aralkoxycarbonyl, carbamoyl, alkylcarbamoyl, and dialkylcarbamoyl, aminoalkyl, aminoalaryl; or a pharmaceutically acceptable salt thereof 2 The method of claim I, wherein the compound has a structure of Formula (H) H 2 wherein: R1 and R2 are independently selected from the group consisting of H, alkyl, aryl, aralkyl, acyl, al koxycarbonyl, aryl oxycarbonyl, aralkoxycarbonyl, carbamoyl, alkylcarbamoyl, and dialkylcarbamoyl, aminoalkyl, aminoalaryl; or a pharmaceutically acceptable salt thereof 3. The method of claim 1, wherein the compound has a structure of Formula (HI) wherein. R1 and R2 are independently selected from the group consisting of H, alkyl, aryl, aralkyl, acyl, a1koxycarbonyl, arOoxycarbonyl, aralkoxycarbonyl, carbamoyl, alkyl carbam oyl, and di al kyl carbam oyl, aminoalkyl, aminoal aryl; or a pharmaceutically acceptable salt thereof 4 The method of claim 1, wherein the compound has a structure of Formula (IV) (IV) The method of claim 1, wherein the compound has a structure of Formula (V) (V) 6 The method of claim 1, wherein the compound has a structure of Formula (VI) y NH (VI) 7. The method of claim 1, wherein the compound has a structure of Formula (VII) (WI) 8. The method of claim 1, wherein the compound has a structure of Formula (VIII) (VIII) 9 A method of treating Parkinson's disease, amyotrophic lateral sclerosis, Alzheimer's disease, drug addiction, traumatic brain injury, psychosis, depression, multiple sclerosis and nausea by administering to the subject one or more compounds, or a pharmaceutically acceptable salt thereof, of claim L