(19) United States (12) Patent Application Publication (10) Pub

US 20070049576A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2007/0049576 A1 Barlow et al. (43) Pub. Date: Mar. 1, 2007

(54) NEUROGENESIS BY MUSCARINIC Related US. Application Data RECEPTOR MODULATION (60) Provisional application No. 60/711,846, ?led on Aug. 26, 2005. Provisional application No. 60/727,127, (75) Inventors: Carrolee Barlow’ Del Mar’ CA (Us); ?led on Oct. 14, 2005. Provisional application No. Tood A_ Carter San Diego CA (Us). 60/738,133, ?led on Nov. 17, 2005. Provisional appli Kym L Lorrain’ San Diego’ CA (USS, cation No. 60/803,826, ?led on Jun. 2, 2006. Jammieson C. Pires, San Diego, CA P bl' t' Cl '? t' (US); Andrew Morse, San Diego, CA u lea Ion assl ca Ion (US); Dana Gitnick, San Marcos, CA (51) Int CL (US); Kai Treuner, San Diego, CA A61]; 31/55 (200601) (US); Alex Broadhead, San Diego, CA A61K 31/473 (200701) (US) A61K 31/4745 (2007.01) A61K 31/445 (2006.01) A61K 31/403 (2007.01) Correspondence Address: A61K 31/66 (2006.01) TOWNSEND AND TOWNSEND AND CREW, (52) US. Cl...... 514/214.03; 514/295; 514/297; LLP 514/319; 514/411; 514/114 TWO EMBARCADERO CENTER EIGHTH FLOOR (57) ABSTRACT SAN FRANCISCO, C A 941114.834 (Us) The instant disclosure describes methods for treating dis eases and conditions of the central and peripheral nervous system by stimulating or increasing neurogenesis. The dis (73) Assignee; Braincells, Inc,’ San Diego, CA closure includes compositions and methods based on mus carinic receptor modulation, such as via inhibition of ace (21) Appl, No.1 11/467,527 tylcholine esterase (AChE) activity, alone or in combination With another neurogenic agent to stimulate or activate the (22) Filed: Aug. 25, 2006 formation of neW nerve cells. Patent Application Publication Mar. 1, 2007 Sheet 1 0f 15 S m A 1

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NEUROGENESIS BY MUSCARINIC RECEPTOR m1 receptors are found in the CNS and peripheral ganglia, MODULATION m2 receptors are found on cardiac cells and in the brainstem, m3 receptors are found in smooth muscle, endocrine (e.g., CROSS-REFERENCES TO RELATED the pancreas) and exocrine glands (e. g., the lacrimal glands), APPLICATIONS and the cerebral cortex, m4 receptors are found primarily in [0001] This application claims bene?t of priority under 35 the neostriatum, and m5 receptors are found primarily in the USC § 119(e) from US. Provisional Patent Applications substantia nigra. Muscarinic receptors are G-protein coupled 60/711,846, ?led Aug. 26, 2005; 60/727,127, ?led Oct. 14, receptors (GPCRs), With the activity of the m1, m3 and m5 2005; 60/738,133, ?led Nov. 17,2005; and 60/803,826, ?led receptors mediated by the phosphoinositide second-messen Jun. 2, 2006; all four of Which are hereby incorporated by ger system, and the m2 and m4 receptors linked to the reference as if fully set forth. adenylate cyclase second messenger system. [0006] Compounds With activity against muscarinic recep FIELD OF THE DISCLOSURE tors have been studied for the treatment of conditions linked [0002] The instant disclosure relates to methods for treat to cholinergic dysfunction, such as AlZheimer’s Disease, ing diseases and conditions of the central and peripheral Which is associated With the degeneration of cholinergic nervous system by stimulating or increasing neurogenesis neurons in the forebrain. Clinical testing has revealed a high via modulation of muscarinic receptor activity, including via degree of debilitating, dose-limiting side effects associated inhibition of acetylcholine esterase (AChE) activity in com With compounds active against the m2 and m3 receptor bination With another neurogenic agent. The disclosure subtypes, including cardiovascular and gastrointestinal side includes methods based on the application of a neurogenesis effects. In contrast, compounds that are selective for m1 modulating agent having activity against muscarinic recep receptors, or m1 and m4 receptors have generally been tors and/ or an AChE inhibitor With another neurogenic agent found to have better clinical pro?les, With enhanced e?icacy to stimulate or activate the formation of neW nerve cells. and decreased side effects. To date, there is has been little research regarding the use of muscarinic compounds to treat BACKGROUND OF THE DISCLOSURE diseases not characteriZed by or substantially resulting from [0003] Neurogenesis is a vital process in the brains of cholinergic dysfunction. animals and humans, Whereby neW nerve cells are continu [0007] Acetylcholinesterase, or AChE, is also knoWn as ously generated throughout the life span of the organism. erythrocyte (or RBC) cholinesterase and acetylcholine The neWly born cells are able to differentiate into functional acetylhydrolase. It is classi?ed at EC 3.1.1.7 and is found in cells of the central nervous system and integrate into exist various locations, including the blood and neural synapses. ing neural circuits in the brain. Neurogenesis is knoWn to AChE catalyZes hydrolysis of the neurotransmitter acetyl persist throughout adulthood in tWo regions of the mamma choline into choline and acetic acid Where acetylcholine is a lian brain: the subventricular Zone (SVZ) of the lateral pan-muscarinic and pan-nicotinic receptor ligand. The ventricles and the dentate gyrus of the hippocampus. In these hydrolysis reaction reaction permits an activated cholinergic regions, multipotent neural progenitor cells (NPCs) continue neuron to return to a resting state. to divide and give rise to neW functional neurons and glial cells (for revieW Gage 2000). It has been shoWn that a [0008] Citation of the above documents is not intended as variety of factors can stimulate adult hippocampal neuro an admission that any of the foregoing is pertinent prior art. genesis, e.g., adrenalectomy, voluntary exercise, enriched All statements as to the date or representation as to the environment, hippocampus dependent learning and anti contents of these documents is based on the information depressants (Yehuda 1989, van Praag 1999, BroWn J 2003, available to the applicant and does not constitute any admis Gould 1999, Malberg 2000, Santarelli 2003). Other factors, sion as to the correctness of the dates or contents of these such as adrenal hormones, stress, age and drugs of abuse documents. negatively in?uence neurogenesis (Cameron 1994, McEWen 1999, Kuhn 1996, Eisch 2004). BRIEF SUMMARY OF THE DISCLOSURE [0004] Muscarinic cholinergic receptors mediate the [0009] Disclosed herein are compositions and methods for effects of the neurotransmitter acetylcholine in the central the prophylaxis and treatment of diseases, conditions and and peripheral nervous systems. In the CNS, muscarinic injuries of the central and peripheral nervous systems by receptors play a central role in mediating cognitive function modulating neurogenesis, including stimulating or increas and are abundant in the forebrain, including the dentate ing the generation of neurons. Aspects of the methods, and gyrus of the hippocampus and the cerebral cortex. In the activities of the compositions, include modulating neuro PNS, muscarinic receptors mediate parasympathetic activ genesis in cases of a disease, disorder, or condition of the ity. Muscarinic receptors are also involved in mediating the nervous system. Embodiments of the disclosure include actions of acetylcholine on certain organs that are responsive methods of treating a neurodegenerative disorder, neurologi to parasympathetic cholinergic stimulation; for example, cal trauma including brain or central nervous system trauma they affect the contractibility of smooth muscle in the and/or recovery therefrom, depression, anxiety, psychosis, gastrointestinal tract, the secretion of gastric acid, the force learning and memory disorders, and ischemia of the central and rate of heart muscle contraction, the secretory activity of and/or peripheral nervous systems. exocrine glands that receive parasympathetic innervation, [0010] In one aspect, methods of modulating, such as by such as the salivary glands, and the constriction of bronchial stimulating or increasing, neurogenesis are disclosed. The tissue. neurogenesis may be at the level of a cell or tissue. The cell [0005] Five sub-types of muscarinic receptors have been or tissue may be present in an animal subject or a human characterized, and are designated m1, m2, m3, m4, and m5. being, or alternatively be in an in vitro or ex vivo setting. In US 2007/0049576 A1 Mar. 1, 2007

some embodiments, neurogenesis is stimulated or increased pharmaceutically acceptable reversible AChE inhibitor. in a neural cell or tissue, such as that of the central or Alternatively, an acceptable irreversible AChE inhibitor may peripheral nervous system of an animal or human being. In also be used in some embodiments of the disclosure. Addi cases of an animal or human, the methods may be practiced tional embodiments comprise an inhibitor that is a tertiary in connection With one or more disease, disorder, or condi amine Which crosses the blood brain barrier. tion of the nervous system as present in the animal or human subject. Thus, embodiments disclosed herein include meth [0016] Non-limiting examples of AChE inhibitors as a muscarinic agent include an organophosphate (such as met ods of treating a disease, disorder, or condition by admin istering at least one neurogenesis modulating agent having rifonate or echothiophate), an aminoacridine (such as activity against muscarinic receptors (hereinafter referred to tacrine), a carbamate (such as physostigmine or neostigmine or rivastigmine) a phenanthrine derivative (such as galan as a “muscarinic agent”). The muscarinic agent may be used tamine), or a piperidine derivative (such as donepeZil) that alone or in combination With one or more additional neu rogenic agents. has neurogenic activity. In other embodiments, a carbamate, such as a quaternary amine (and pyridostigmine, [0011] While a muscarinic agent may be considered a ambenonium and demarcarium as non-limiting examples) “direct” agent in that it has direct activity against a musca are disclosed for use in the practice of the invention. Other rinic receptor by interactions thereWith, the disclosure non-limiting examples of an AChE inhibitor With neuro includes a muscarinic agent that may be considered an genic activity include itopride, (—)-huperZine A, and “indirect” agent in that it does not directly interact With a phenserine. Ayet additional embodiment of anAChE inhibi muscarinic receptor. Thus, an indirect agent acts on a tor is edrophonium. muscarinic receptor indirectly, or via production, generation, [0017] Embodiments described herein include a combina stability, or retention of an intermediate agent Which directly tion of more than one of the muscarinic agents disclosed interacts With a muscarinic receptor. In other embodiments, herein or knoWn to the skilled person. Of course a neuro an indirect agent may be one that increases the amount of genic muscarinic agent may be used, either alone or in choline or available choline, such as by increasing choline combination With one or more additional muscarinic agents synthesis as a non-limiting example. or other neurogenic agents. Compositions disclosed herein [0012] In some embodiments, an indirect agent is an include such combinations of muscarinic agents, including inhibitor of AChE activity. Like a direct muscarinic agent, an AChE inhibitor(s) as a non-limiting embodiment, and other indirect agent may be used alone or in combination With one neurogenic agent(s). or more additional neurogenic agents, as described herein. [0018] In another aspect, the disclosed methods include An AChE inhibitor may be a cholinergic agonist, such as an inhibitor that suppresses the enZymatic activity of AChE. In identifying a patient suffering from one or more diseases, disorders, or conditions, or a symptom thereof, and admin some embodiments, an AChE inhibitor binds an AChE istering to the patient a muscarinic agent, alone or in active site. combination With another neurogenic agent, as described [0013] An additional neurogenic agent as described herein herein. In some embodiments, a method including identi? may be another direct muscarinic agent, another indirect cation of a subject as in need of an increase in neurogenesis, muscarinic agent (such as AChE inhibitor), or a neurogenic and administering to the subject one or more muscarinic agent that does not act, directly or indirectly, through a agents is disclosed herein. In other embodiments, the subject muscarinic receptor. Thus in some embodiments, an addi is a patient, such as a human patient. tional neurogenic agent is one that acts, directly or indirectly, through a mechanism other than a muscarinic receptor. [0019] Additional embodiments describe a method includ ing administering a muscarinic agent, alone or in combina [0014] In one aspect, methods of modulating, such as by tion With another neurogenic agent, to a subject exhibiting stimulating or increasing, neurogenesis are disclosed. The the effects of insuf?cient amounts of, or inadequate levels of, neurogenesis may be at the level of a cell or tissue. The cell neurogenesis. In some embodiments, the subject may be one or tissue may be present in an animal subject or a human that has been subjected to an agent that decreases or inhibits being, or alternatively be in an in vitro or ex vivo setting. In neurogenesis. Non-limiting examples of an inhibitor of some embodiments, neurogenesis is stimulated or increased neurogenesis include opioid receptor agonists, such as a mu in a neural cell or tissue, such as that of the central or receptor subtype agonist like morphine. In a related manner, peripheral nervous system of an animal or human being. In a method provides for administering a muscarinic agent, cases of an animal or human, the methods may be practiced alone or in combination With another neurogenic agent, to a in connection With one or more disease, disorder, or condi subject or person that Will be subjected to an agent that tion of the nervous system as present in the animal or human decreases or inhibits neurogenesis. Non-limiting embodi subject. Thus, embodiments disclosed herein include meth ments include those Where the subject or person is about to ods of treating a disease, disorder, or condition by admin be administered morphine or another opioid receptor ago istering a muscarinic agent alone or in combination With nist, like another opiate, and so about to be subject to a another neurogenic agent, as described herein. The addi decrease or inhibition of neurogenesis. Non-limiting tional neurogenic agent may be another muscarinic agent examples include administering a muscarinic agent, alone or (direct or indirect) or a neurogenic agent that acts through a in combination With another neurogenic agent, to a subject mechanism independent from a muscarinic receptor. before, simultaneously With, or after the subject is admin istered morphine or other opiate in connection With a [0015] In another aspect, methods of using chemical enti surgical procedure. ties as muscarinic agents to modulate, such as by increasing, neurogenesis are disclosed. In some embodiments, a chemi [0020] Also disclosed are methods for preparing a popu cal entity used as a muscarinic agent is a therapeutically or lation of neural stem cells suitable for transplantation, com US 2007/0049576 A1 Mar. 1, 2007

prising culturing a population of neural stem cells (NSCs) in neural stem cells (hNSC) in neurosphere culture (diamonds: vitro, and contacting the cultured neural stem cells With at 1 uM sabcomeline; squares: l0 uM sabcomeline; triangles: least one muscarinic agent, optionally in combination With 30 uM sabcomeline, X: positive control (basal media With another muscarinic agent, such as an AChE inhibitor, and/or knoWn proliferative agent); asterisk: negative control (basal another neurogenic agent. In some embodiments, the stem media). Neurogenesis is measured as the total area of a cells are prepared and then transferred to a recipient host single neurosphere comprising human neural stem cells as a animal or human. Non-limiting examples of preparation function of time. Results are presented as the percent include 1) contact With a muscarinic agent, optionally in increase in area over basal media. Sabcomeline enhanced combination With another muscarinic agent, such as an proliferation and/or survival of hNSC compared to basal AChE inhibitor, and/or another neurogenic agent, until the media at all time periods tested. cells have undergone neurogenesis, such as that Which is [0027] FIG. 5 is a series of immuno?uorescent micro detectable by visual inspection or cell counting, or 2) contact scopic images of monolayers of human neural stem cells With a muscarinic agent, such as an AChE inhibitor, option (hNSC) after immunohistochemistry staining With the neu ally in combination With another muscarinic agent and/or ronal marker TUJ-l (blue), the astrocyte marker GFAP (red), another neurogenic agent, until the cells have been suffi and a nuclear cell marker (Hoechst 33342) (example 5). The ciently stimulated or induced toWard or into neurogenesis. upper left image is a negative control (basal media), the The cells prepared in such a non-limiting manner may be upper middle image is a neuronal positive control (basal transplanted to a subject, optionally With simultaneous, media plus a knoWn promoter of neuronal differentiation), nearly simultaneous, or subsequent administration of a neu and the upper right image is an astrocyte positive control rogenic agent, or a muscarinic agent, such as an AChE (basal media plus a knoWn promoter of astrocyte differen inhibitor, to the subject. While the neural stem cells may be tiation). The loWer image shoWs the effect of sabcomeline on in the form of an in vitro culture or cell line, in other hNSC differentiation. embodiments, the cells may be part of a tissue Which is subsequently transplanted into a subject. [0028] FIG. 6 shoWs the effect of chronic dosing With sabcomeline on the locomotor activity of rats (squares: 0.01 [0021] In yet another aspect, the disclosure includes meth mg/kg; triangles: 0.05 mg/kg; diamonds: negative control ods of modulating, such as by stimulating or increasing, (vehicle)). Sabcomeline had no signi?cant effect on loco neurogenesis in a subject by administering a muscarinic motor activity. “mpk” stands for mg/kg. agent, optionally in combination With another muscarinic agent, such as anAChE inhibitor, and/or another neurogenic [0029] FIG. 7 shoWs the effect of chronic dosing With agent. In some embodiments, the neurogenesis occurs in sabcomeline on the body Weight of rats (squares: 0.01 combination With the stimulation of angiogenesis Which mg/kg; triangles: 0.05 mg/kg; diamonds: negative control provides neW cells With access to the circulatory system. (vehicle)). Sabcomeline had no signi?cant effect on body Weight. [0022] The details of additional embodiments are set forth in the accompanying draWings and the description beloW. [0030] FIG. 8 shoWs the effect of acute dosing With Other features, objects, and advantages of the embodiments sabcomeline on the performance of rats in the modi?ed Will be apparent from the draWings and detailed description, forced sWim test (dark grey: 0.01 mg/kg; White: 0.05 mg/kg; and from the claims. light grey: negative control (vehicle). Sabcomeline had no signi?cant effects on performance in the sWim test. BRIEF DESCRIPTION OF THE DRAWINGS [0031] FIG. 9 shoWs the effect of chronic dosing of rats With sabcomeline on latency to eat in a novelty-suppressed [0023] FIG. 1 is a dose-response curve shoWing effect of feeding assay (left: vehicle; middle: 0.01 mg/kg; right: 0.05 the muscarinic agent sabcomeline on neuronal differentia mg/kg). Sabcomeline decreased the latency to eat in a tion. Data is presented as the percentage of the neuronal dose-dependent manner, With a statistically signi?cant positive control, With basal media values subtracted. EC5O decrease at 0.05 mg/kg. Was observed at a sabcomeline concentration of 8.04 uM, compared to 5.28 uM for the positive control. [0032] FIG. 10 shoWs the effect of chronic dosing of rats With sabcomeline on cognitive performance in a novel [0024] FIG. 2 is a dose-response curve shoWing effect of object recognition assay (left: vehicle; middle: 0.01 mg/kg; the muscarinic agent sabcomeline on astrocyte differentia right: 0.05 mg/kg). Sabcomeline signi?cantly enhanced per tion. Data is presented as the percentage of the astrocyte formance in the assay at 0.01 mg/kg. positive control, With basal media values subtracted. Sab comeline Was found to have no effect on astrocyte differ [0033] FIG. 11 shoWs the effect of acute dosing of rats entiation. (injection once daily for ?ve days) With sabcomeline on neural cell proliferation Within the dentate gyrus (left: [0025] FIG. 3 is a dose-response curve measuring the vehicle; middle: 0.01 mg/kg; right: 0.05 mg/kg). Results are toxicity/trophism effect of the muscarinic agent sabcomeline presented as the mean number of Brdu-positive cells per on a population of cultured neural stem cells. Data is cubic mm. A dose-related increase in proliferation Was presented as the percentage of the basal media cell count. observed. Toxic doses produce viability values beloW 80% of the basal media control, While trophic compounds shoW a dose [0034] FIG. 12 shoWs the effect of chronic dosing of rats dependent increase in cell count. Sabcomeline has no detect With Sabcomeline on the differentiation of neural progenitor able toxicity, and shoWs a signi?cant trophic effect. cells into mature neuron Within the subgranular Zone of the dentate gyrus. Chronic Sabcomeline treatment resulted in a [0026] FIG. 4 shoWs the effect of various concentrations of 20 and 18 percent increase at 0.01 and 0.05 mg/kg/day, the muscarinic agent sabcomeline on neurogenesis of human respectively. US 2007/0049576 A1 Mar. 1, 2007

[0035] FIG. 13A shows the effect of chronic dosing ofrats [0039] A “neurogenic agent” is de?ned as a chemical With sabcomeline, ?uoxetine, or a combination of both agent or reagent that can promote, stimulate, or otherWise agents in the novelty suppressed feeding depression model increase the amount or degree or nature of neurogenesis in (gray: vehicle; White: 5.0 mg/kg ?uoxetine; hatched: 0.01 vivo or ex vivo or in vitro relative to the amount, degree, or mg/kg sabcomeline; cross-hatched: 5.0 mg/kg Fluoxetine nature of neurogenesis in the absence of the agent or reagent. and 0.01 mg/kg sabcomeline). The combination of ?uoxet In some embodiments, treatment With a neurogenic agent ine With sabcomeline resulted in signi?cantly enhanced increases neurogenesis if it promotes neurogenesis by at performance in the assay relative to vehicle or either agent least about 5%, at least about 10%, at least about 25%, at alone. FIG. 13B shoWs the effect of acute dosing of rats least about 50%, at least about 100%, at least about 500%, (injection once daily for ?ve days) With sabcomeline, ?u or more in comparison to the amount, degree, and/or nature oxetine, or a combination of both agents on neural cell of neurogenesis in the absence of the agent, under the proliferation Within the dentate gyrus (gray: vehicle; White: conditions of the method used to detect or determine neu 5.0 mg/kg ?uoxetine; hatched: 0.01 mg/kg sabcomeline; rogenesis. As described herein, a muscarinic agent, such as cross-hatched: 5.0 mg/kg Fluoxetine and 0.01 mg/kg sab an AChE inhibitor, that promotes, stimulates, or otherWise comeline) in the dorsal hippocampus and the ventral hip increases the amount or degree or nature of neurogenesis is pocampus. Results are presented as the mean number of a neurogenic agent. Brdu-positive cells per cubic mm. The combination of sabcomeline and ?uoxetine resulted in a signi?cant increase [0040] The term “astrogenic” is de?ned in relation to in proliferation indicating increased neurogenesis. “astrogenesis” Which refers to the activation, proliferation, differentiation, migration and/or survival of an astrocytic [0036] FIG. 14 is a dose-response curve shoWing effects of cell in vivo or in vitro. Non-limiting examples of astrocytic the neurogenic agents donepeZil (acetylcholinesterase cells include astrocytes, activated microglial cells, astrocyte inhibitor) and captopril (angiotensin converting enZyme precursors and potentiated cells, and astrocyte progenitor inhibitor) in combination on neuronal differentiation com and derived cells. In some embodiments, the astrocyte is an pared to the effect of either agent alone. When run indepen adult, fetal, or embryonic astrocyte or population of astro dently, each compound Was tested in a concentration cytes. The astrocytes may be located in the central nervous response curve ranging from 0.01 uM to 31.6 uM. In system or elseWhere in an animal or human being. The combination, the compounds Were combined at equal con astrocytes may also be in a tissue, such as neural tissue. In centrations at each point (for example, the ?rst point in the some embodiments, the astrocyte is an adult, fetal, or combined curve consisted of a test of 0.01 uM donepeZil and embryonic progenitor cell or population of cells, or a popu 0.01 uM captopril). Data is presented as the percentage of lation of cells comprising a mixture of stem and/or progeni the neuronal positive control, With basal media values sub tor cells, that is/are capable of developing into astrocytes. tracted. When used alone, EC5O Was observed at a donepeZil Astrogenesis includes the proliferation and/or differentiation concentration of 4.7 uM or a captopril concentration of 2.0 of astrocytes as it occurs during normal development, as uM in test cells. When used in combination neurogenesis is Well as astrogenesis that occurs folloWing disease, damage greatly enhanced: EC5O Was observed at a combination of or therapeutic intervention. donepeZil and captopril at concentrations of 0.24 uM each. [0041] The term “stem cell” (or neural stem cell (N SC)), [0037] FIG. 15 is a dose-response curve shoWing the as used herein, refers to an undifferentiated cell that is reduction of astrocyte differentiation by buspirone (a capable of self-reneWal and differentiation into neurons, 5-HT1a agonist) in combination With donepeZil (an acetyl astrocytes, and/or oligodendrocytes. cholinesterase inhibitor). When run alone, buspirone potently induces astrocyte differentiation With an EC5O of [0042] The term “progenitor cell” (e.g., neural progenitor 5.7 uM at a maximum of 60% of the positive control. The cell), as used herein, refers to a cell derived from a stem cell addition of donepeZil results in signi?cant inhibition of that is not itself a stem cell. Some progenitor cells can astrocyte differentiation to a maximum of 14% positive produce progeny that are capable of differentiating into more control With an estimated EC5O greater than 31.6 uM. than one cell type. DETAILED DESCRIPTION OF MODES OF [0043] The term “muscarinic agent” as used herein PRACTICE includes a neurogenic agent, as de?ned herein, that elicits an [0038] “Neurogenesis” is de?ned herein as proliferation, observable response upon contacting a muscarinic receptor. differentiation, migration and/or survival of a neural cell in “Muscarinic agents” useful in the methods described herein vivo or in vitro. In various embodiments, the neural cell is include compounds or agents that, under certain conditions, an adult, fetal, or embryonic neural stem cell or population may act as: agonists (i.e., agents able to elicit one or more of cells. The cells may be located in the central nervous biological responses of a muscarinic receptor); partial ago system or elseWhere in an animal or human being. The cells nists (i.e., agents able to elicit one or more biological may also be in a tissue, such as neural tissue. In some responses of a receptor to a less than maximal extent, e.g., embodiments, the neural cell is an adult, fetal, or embryonic as de?ned by the response of the receptor to an agonist); progenitor cell or population of cells, or a population of cells antagonists (agents able to inhibit one or more characteristic comprising a mixture of stem cells and progenitor cells. responses of a muscarinic receptor, for example, by com Neural cells include all brain stem cells, all brain progenitor petitively or non-competitively binding to the muscarinic cells, and all brain precursor cells. Neurogenesis includes receptor, a ligand of the receptor, and/or a doWnstream neurogenesis as it occurs during normal development, as signaling molecule); and/or inverse agonists (agents able to Well as neural regeneration that occurs folloWing disease, block or inhibit a constitutive activity of a muscarinic damage or therapeutic intervention, such as by the treatment receptor) of one or more subtypes of muscarinic receptors. described herein. For example, the muscarinic agent alvameline, described in