WO2010009453A1 - Methods and systems for evaluating memory agents - Google Patents
Methods and systems for evaluating memory agents Download PDFInfo
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- WO2010009453A1 WO2010009453A1 PCT/US2009/051082 US2009051082W WO2010009453A1 WO 2010009453 A1 WO2010009453 A1 WO 2010009453A1 US 2009051082 W US2009051082 W US 2009051082W WO 2010009453 A1 WO2010009453 A1 WO 2010009453A1
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/15—Medicinal preparations ; Physical properties thereof, e.g. dissolubility
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
- A61K31/445—Non condensed piperidines, e.g. piperocaine
- A61K31/451—Non condensed piperidines, e.g. piperocaine having a carbocyclic group directly attached to the heterocyclic ring, e.g. glutethimide, meperidine, loperamide, phencyclidine, piminodine
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
- A61K31/4965—Non-condensed pyrazines
- A61K31/497—Non-condensed pyrazines containing further heterocyclic rings
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
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- C—CHEMISTRY; METALLURGY
- C40—COMBINATORIAL TECHNOLOGY
- C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
- C40B30/00—Methods of screening libraries
- C40B30/06—Methods of screening libraries by measuring effects on living organisms, tissues or cells
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
Definitions
- the invention relates to methods and systems of evaluating, identifying or assessing the effectiveness of memory agent and/or training protocols administered to subjects.
- Memory is the recollection of past experiences. One is able to register new information, store it and then recall it sometime later. Recollection of facts and events represents a form of conscious, or explicit, memory. One also remembers less conscious skills, such as riding a bike, playing a musical instrument or associating simple cues with reward or punishment, which represent more implicit forms of memory. For both of these cases, the process of memory formation appears to proceed through three general stages. Acquisition (learning) involves the initial perception of a new experience. Short-term memory of this newly acquired experience then appears transient and labile. Under appropriate conditions, often requiring repetition and rest, short-term memory of an experience then can be "consolidated" into a more durable long- term memory. An every-day example of this memory process involves using a phone number.
- the human brain is evolutionarily designed to sense its internal and external environment, to perceive causal relations among stimuli and to change its response adaptively. To accomplish this enormous task, a self-regulating network of 40 thousand genes directs the development of a cellular network of one hundred billion neurons with 100 trillion connections among them. Such complexity is staggering. Yet neurobiologists have begun to understand how the human brain works in part by studying brain function in other animals.
- ASM anesthesia-sensitive memory
- LTM long-term memory
- CREB gene is expressed in two opposing forms - an activator or a repressor of downstream, CRE -mediated gene transcription.
- Key behavioral experiments in Drosophila originally established a critical role for CREB in the formation of long-term memory.
- Normal fruit flies are capable of learning the association between a particular smell and the punishment of foot shock. After one training session, a majority of flies will avoid the previously shock-paired odor when given a choice between it and a neutral odor in a T-maze. Though such learning initially is quite robust, memory thereafter is transient, decaying away within a day.
- a bonafide protein synthesis-dependent LTM lasting more than a week can be produced in normal flies when subjected to ten training sessions with a 15-min rest interval between them.
- LTM specifically is blocked with no effects on learning or early memory.
- flies genetically engineered to produce high levels of CREB activator LTM after spaced training is not affected but, surprisingly, is induced after just one training session.
- overexpression of CREB activator enhances memory by reducing the number of training sessions required to induce the formation of LTM — the functional equivalent of a photographic memory.
- CREB Several intracellular signaling pathways appear to be induced initially by neural activity, but CREB nonetheless appears to be a dominant "downstream" target, regulating the conversion of early memory to lasting memory. Down-regulation of CREB suppresses, while up-regulation of CREB enhances, a gene transcriptional response underlying a synaptic grow process that yields lasting functional and structural plasticity at synapses. In essence, CREB appears to act as a master switch that determines when neural activity will give rise to lasting, structural changes in a circuit. A key factor influencing this switch is repetition; when the CREB pathway is up- regulated, the number of training sessions required for the formation of long-term memory is decreased.
- the screen needed to identify drugs that had no effect on CREB function alone but rather synergized with co-activation of cAMP signaling. This requirement was designed to mimic the original behavioral experiments. In flies, overexpression of CREB alone had no effect on fly behavior. Rather, training per se was required to produce enhanced memory, because experience-induced increases in neural activity activated the cAMP pathway.
- HTS high throughput screen
- human neuroblastoma cells which were stably transfected with a CRE-luciferase reporter gene.
- changes in CREB-dependent transcription of luciferase could be monitored via fluorescence.
- drugs were sought that produced no change in luciferase activity on their own but synergized (> 2-fold) with a suboptimal dose of forskolin, which activates the cAMP pathway (as does neural activity in vivo).
- PDE inhibitor decreasing PDE activity leads to an increase of cAMP normally synthesized by adenylyl cyclase (AC), and elevated cAMP leads to activation of CREB via PKA.
- a PDE inhibitor might be expected to have minimal effect alone, in the absence of forskolin-induced activation of AC (or neural activity-induced activation of AC in vivo).
- HT-0712 An advanced PDE-4 inhibitor, HT-0712, has been shown to be a validated compound for clinical development, along with several other CREB pathway enhancers in addition to HT-0712. To date, behavioral assessments in rodent models of memory formation and biochemical experiments have shown that these compounds effectively enhance memory.
- Such therapies may include particular training protocols and/or administration of such compound or compounds, including compounds that serve as memory agents, which may be memory enhancers that specifically up-regulate the CREB pathway, as well as memory impairers.
- the present invention includes methods including the steps of assessing the effectiveness of a memory agent administered to a subject, wherein said assessing comprises presenting to the subject sets of stimuli and evaluating the subject's response to the stimuli.
- the assessing may include a first period comprising presenting to the subject stimuli associated into sets and evaluating the subject's response to the stimuli to establish a criterion; and a second period comprising presenting to the subject stimuli associated into sets and evaluating the subject's response to the stimuli to establish a long term success measurement.
- the second period begins at least about one day after the completion of the first period.
- the evaluating of the subject's response may include comparing the criterion to the long term success measurement.
- the sets of stimuli may include a plurality of pairs of stimuli.
- the each of the plurality of pairs of stimuli can include a positive element and/or a negative element.
- each of the plurality of pairs of stimuli may include an element that has an identifiable association with the other element of the pair. In some embodiments, this identifiable association may be a face - name association or word - word association.
- the subject may be an animal. In certain embodiments, the animal may be a primate, a mammal, a mouse or a rat. In some embodiments, the primate may be a human, a monkey, a lemur, a macaque or an ape. In some embodiments, the first period may be from about one to about ten days.
- the second period may be about one day.
- there may be a plurality of subjects which may include a control group and an experimental group.
- the assessing may include comparing the control group to the experimental group.
- the evaluating of the subject's response includes calculating and/or measuring one or more aspects selected from the group consisting of: the ratio of the long term success measurement to the criterion, long term memory retention, the amount of time required to achieve a particular criterion, and/or the amount of time between the first period and the second period while keeping a constant, near-constant, or improved ratio of the long term success measurement to the criterion.
- the present invention may involve a method including selecting a memory agent as a drug candidate, wherein said selecting includes the steps of: administering a memory agent to a subject; and presenting to the subject sets of stimuli and evaluating the subject's response to the stimuli.
- the assessing may further include: a first period comprising presenting to the subject stimuli associated into sets and evaluating the subject's response to the stimuli to establish a criterion; and a second period comprising presenting to the subject stimuli associated into sets and evaluating the subject's response to the stimuli to establish a long term success measurement.
- the second period begins at least about one day after the completion of the first period.
- the evaluating of the subject's response comprises calculating and/or measuring one or more aspects selected from the group consisting of: the ratio of the long term success measurement to the criterion, long term memory retention, the amount of time required to achieve a particular criterion, and/or the amount of time between the first period and the second period while keeping a constant, near-constant, or improved ratio of the long term success measurement to the criterion.
- the sets of stimuli comprise a plurality of pairs of stimuli.
- the subject may be an animal, including an mammal such as a primate, a mouse and a rat.
- the present invention involves a method including: testing a memory agent as a long term memory enhancer, wherein said testing includes the steps of: administering a memory agent to a subject; and presenting to the subject sets of stimuli and evaluating the subject's response to the stimuli.
- the assessing further includes: a first period comprising presenting to the subject stimuli associated into sets and evaluating the subject's response to the stimuli to establish a criterion; and a second period comprising presenting to the subject stimuli associated into sets and evaluating the subject's response to the stimuli to establish a long term success measurement.
- the second period begins at least about one day after the completion of the first period.
- the evaluating of the subject's response comprises calculating and/or measuring one or more aspects selected from the group consisting of: the ratio of the long term success measurement to the criterion, long term memory retention, the amount of time required to achieve a particular criterion, and/or the amount of time between the first period and the second period while keeping a constant, near-constant, or improved ratio of the long term success measurement to the criterion.
- the sets of stimuli comprise a plurality of pairs of stimuli.
- the subject may be an animal, including an animal selected from the group consisting of a primate, a mammal, a mouse and a rat.
- the present invention may be a method including: assessing the effectiveness of a memory agent, wherein said assessing comprises the steps of: providing a first subject and a second subject, wherein the first subject and the second subject are of the same species; administering a memory agent to the first subject; presenting the first subject and the second subject a plurality of pairs of stimuli during a first period and evaluating the first subject and the second subject's responses to the stimuli to establish a criterion for each of the first subject and the second subject; presenting the first subject and the second subject the plurality of pairs of stimuli during a second period and evaluating the first subject and the second subject's responses to the stimuli to establish a long term success measurement for each of the first subject and the second subject; comparing the first subject and the second subject's criterions to their respective long term success measurements.
- the second period begins at least about one day after the completion of the first period. In some embodiments, the second period begins from about 1 to about 10 days following the completion of the first period.
- the comparing of the criterions to the long term success measurements comprises: establishing a first ratio of the first subject's long term success measurement to the first subject's criterion; establishing a second ratio of the second subject's long term success measurement to the second subject's criterion; and comparing the first ratio to the second ratio.
- the sets of stimuli include a plurality of pairs of stimuli. In some embodiments, the plurality of pairs of stimuli comprises a positive element and/or a negative element.
- each of the plurality of pairs of stimuli comprises an element that has an identifiable association with the other element.
- the identifiable association is selected from the group consisting of a face - name association and a word - word association.
- each of the first subject and the second subject is an animal, including an animal is selected from the group consisting of a primate (including a human, a monkey, a lemur, a macaque and an ape), a mammal, a mouse and a rat.
- the first period comprises from about one to about ten days.
- the second period comprises about one day.
- each of the first subject and the second subject is a plurality of subjects.
- the present invention includes methods similar to those recited above, but which focus on evaluating an training protocol rather than or in addition to a memory agent.
- Some embodiments involve a method including the steps of: assessing the effectiveness of an training protocol administered to a subject, wherein said assessing comprises presenting to the subject sets of stimuli and evaluating the subject's response to the stimuli.
- Other embodiments involve a method including: testing a training protocol as a long term memory enhancer, wherein said testing includes the steps of: administering an training protocol to a subject; and presenting to the subject sets of stimuli and evaluating the subject's response to the stimuli.
- Further embodiments involve a method including: assessing the effectiveness of an training protocol, wherein said assessing includes the steps of: providing a first subject and a second subject, wherein the first subject and the second subject are of the same species; administering an training protocol to the first subject; presenting the first subject and the second subject a plurality of pairs of stimuli during a first period and evaluating their responses to the stimuli to establish a criterion for each of the first subject and the second subject; presenting the first subject and the second subject the plurality of pairs of stimuli during a second period and evaluating their responses to the stimuli to establish a long term success measurement for each of the first subject and the second subject; comparing the criterions of the first subject and the second subject to their respective long term success measurements.
- the present invention also includes systems, including systems for use along with the methods disclosed herein.
- the system may include all elements necessary to perform an embodiment of the methods of the present invention.
- Systems of the present invention may be useful for selecting a memory agent as a drug candidate, assessing the effectiveness of a memory agent, assessing the effectiveness of a training protocol, and/or testing a memory agent as a long term memory enhancer, and/or testing a memory agent along with an training protocol as a long term memory enhancer.
- a system of the present invention includes sets of stimuli, wherein said sets of stimuli are configured to permit evaluation of a subject's response to the sets of stimuli and thereby assess the effectiveness of an training protocol administered to the subject.
- the sets of stimuli comprise a plurality of pairs of stimuli.
- each of the plurality of pairs of stimuli comprises a positive element and/or a negative element.
- each of the plurality of pairs of stimuli comprises an element that has an identifiable association with the other element of the pair.
- the identifiable association is selected from the group consisting of a face - name association and a word - word association.
- the system may further include testing protocols, testing schedules, and/or memory agent dosage and administration schedules.
- a system of the present invention includes sets of stimuli, wherein said sets of stimuli are configured to permit evaluation of a subject's response to the sets of stimuli and thereby assess the effectiveness of a memory agent administered to the subject.
- the sets of stimuli comprise a plurality of pairs of stimuli.
- each of the plurality of pairs of stimuli comprises a positive element and/or a negative element.
- each of the plurality of pairs of stimuli comprises an element that has an identifiable association with the other element of the pair.
- the identifiable association is selected from the group consisting of a face - name association and a word - word association.
- the system may further include testing protocols, testing schedules, and/or memory agent dosage and administration schedules.
- the methods and systems of the present invention utilize a memory agent that is one or more selected from the group consisting of a compound having the structure:
- each of Yi, Y 2 , Y 3 , and Y 4 is independently — H; straight chained or branched C 1 - C 7 alkyl, monofluoroalkyl or polyfluoroalkyl; straight chained or branched C 2 -C 7 alkenyl or alkynyl; C 3 -C 7 cycloalkyl or C 5 -C 7 cycloalkenyl; -F, -Cl, -Br, or -I; -NO 2 ; -N 3 ; -CN; -OR 4 , -SR 4 , -OCOR 4 , -COR 4 , -NCOR 4 , -N(R-O 2 , -CON(R-O 2 , or -COOR 4 ; aryl or heteroaryl; or any two of Yi, Y 2 , Y 3 and Y 4 present on adjacent carbon atoms can constitute a methylenedioxy group; wherein each
- Ri and R 2 are each independently H, straight chained or branched Ci-C 7 alkyl, - F, -Cl, —Br, —I, -NO 2 , or -CN; wherein R 3 is H, straight chained or branched Ci-C 7 alkyl, — F, -Cl, -Br, —I, -NO 2 , -CN, -OR 6 , aryl or heteroaryl; wherein R 5 is straight chained or branched Ci-C 7 alkyl, — N(R-O 2 , -OR 4 or aryl; wherein R 6 is straight chained or branched Ci-C 7 alkyl or aryl; wherein B is aryl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, indolizinyl, indol- 4-yl, indol-5-yl,
- the methods and systems of the present invention utilize a memory agent that is one or more selected from the group consisting of a compound having the structure:
- Me is methyl
- cPent is cyclopentyl and the 3 and 5 carbons are in the S configuration.
- FIGS. IA - IB illustrate graphs showing results from an embodiment of the present invention described in Example 1. Results are expressed as mean days ⁇ s.e.m per treatment, * p ⁇ 0.05 versus vehicle.
- FIGS. 2 A - 2B illustrate graphs showing results from an embodiment of the present invention described in Example 1. Results are expressed as mean percent performance ⁇ s.e.m. per treatment.
- FIGS. 3 A - 3B illustrate graphs showing results from an embodiment of the present invention described in Example 1. Results are expressed as mean percent performance ⁇ s.e.m. per treatment.
- FIGS. 4A - 4D illustrate exemplary pairs of visual stimuli that may be used in an embodiment of the present invention described in Example 1.
- FIG. 5 illustrates a graph showing performance of healthy elderly volunteers in a paired- associate (face-name) memory assay embodiment described in Example 2.
- Subjects were asked to memorize a set of 40 pairs of faces and names (pro professions) for 6 days (d2 - d7). On training days, subjects went through the list of 40 pairs only once. They were queried for their recall first and then given the correct answers to continue training. After training, three-day memory retention was quantified using only face-name pairs 1 - 20 (dlO). Seven-day memory retention was quantified again for these face-name pairs 1 - 20 (dl4b). Seven-day retention also was quantified for face-name pairs 21 - 40 twice (dl4a and dl4a2), with a 1-min interval between recall sessions.
- the present invention provides methods and systems for identifying, evaluating, and testing various compounds as memory agents and regimens as training protocols. Such methods and systems may test any compounds or protocols, including the memory agents and training protocols described herein. The methods can comprise combining training protocols with the general administration of memory agents. In some embodiments, the present invention may involve identifying, selecting, testing, evaluating or assessing a compound as a drug candidate for a memory agent, for example, as one effective for enhancing memory.
- a drug candidate may be a compound that has been identified through a drug discovery process and has been synthesized, characterized, optimized, and/or screened or assayed. Once a compound has been shown to have potential for a given activity or effect, for example by use of the methods or systems of the present invention, it may be a drug candidate - i.e., one that may be put through clinical trials with the intent that it be ultimately commercialized.
- the present invention involves the use of certain tasks to assess one or more aspects of an animal's memory capabilities. For example, the animal's learning capabilities, short term memory, or long term memory retention may be tested and/or evaluated.
- the term "memory,” without an additional modifier (e.g., short term memory) refers to long term memory.
- the term “memory agent” refers to memory augmenting agents or memory suppressing agents. Memory augmenting and suppressing agents include, but are not limited to, those molecules, compounds, and/or agents that act on a CREB pathway, as described, for example, in U.S. Patent Nos. 5,929,223, 6,051,559, 6,689,557, and 6,890,516, and in U.S. Patent Application Serial No. 11/066,125, the disclosure of each of which is incorporated by reference in their entirety herein.
- animal includes mammals, as well as other animals, vertebrate, invertebrate (e.g., birds, fish, reptiles, insects (e.g., Drosophila species) and mollusks (e.g., Aplysia)).
- vertebrate invertebrate
- insects e.g., Drosophila species
- mollusks e.g., Aplysia
- mammal and “mammalian” refer to any vertebrate animal, including monotremes, marsupials and placental, that suckle their young and either give birth to living young (eutharian or placental mammals) or are egg-laying (metatharian or nonplacental mammals).
- mammalian species include, without limitation, humans and primates (e.g., monkeys, chimpanzees, humans, apes, lemurs, or macaques), rodents (e.g., rats, mice, guinea pigs) and ruminents (e.g., cows, pigs, horses).
- primates e.g., monkeys, chimpanzees, humans, apes, lemurs, or macaques
- rodents e.g., rats, mice, guinea pigs
- ruminents e.g., cows, pigs, horses.
- the animal can be an animal with some form and degree of memory dysfunction or an animal with normal memory performance (i.e., an animal without any form of memory failure (dysfunction or loss of any memory)).
- One or more animals may be used and such animals may be placed in one or more groups (e.g., one or more control groups and/or one or more experimental groups).
- the animal may be any suitable age.
- the tasks may involve the presentation of certain stimuli to the subjects.
- the stimuli may be any suitable sensory stimulus including, without limitation, auditory, visual, tactile, olfactory, gustatory and/or combinations thereof.
- the stimuli may be visual and be one or more of a symbol, letter, number, line, shape, color, picture or combinations thereof.
- the stimuli may be associated into sets, such that a certain relationship is established between the members of the sets.
- the stimuli may be associated into groups of about 2, 3, 4, 5, 6, 7, 8, 9 or 10.
- the stimuli are associated into pairs.
- each stimulus in the pair is of the same type (i.e., audio, visual or tactile) while in other embodiments the pair may be of different types.
- an element in the pair will have some identifiable association with the other element of the pair.
- An identifiable association can be a visual, audible, logical or other aspect of the one or more elements of the pair that would cause one to associate that element of the pair with its corresponding element.
- the pair could be a word-word pair wherein one word does not have an identifiable association with the other word (e.g., comb-apple).
- An example of visual paired stimuli is depicted in FIGs. 4A-4D.
- Another example of a stimulus pair with an identifiable association could be a face - name pairing and/or an occupation - face/name pairing. In some embodiments, there may also be an occupation listed along with the face - name pairing.
- the subjects may be presented one or more pairs of stimuli.
- 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 50, 100, 120, 150, or any other suitable number of pairs of stimuli may be presented to the subject.
- one element of the pair is designated to be positive - meaning that the subject will receive a reward if it selects that element.
- one element of the pair is designated to be negative - meaning that the subject will receive negative feedback if it selects that element.
- the subject selects the element by touching the element, but in other embodiments any other suitable selection method may be used.
- the reward may be food, but in other embodiments the reward may be a pleasant sound, access to certain toys or items, or anything suitable for triggering a positive response from the subject.
- the negative feedback is a shock, a sound, a punishment or withholding of privileges, or any suitable action or the like that provides the subject with negative feedback.
- the type of feedback, the selection of the pairs of stimuli, sets of pairs of stimuli, species and age of the subjects, and other aspects of the tasks may be modified to test and/or evaluate different aspects of a subject's memory. For example, using stimulus pairs wherein each element has an identifiable association to the other element may be used to evaluate the subject's relational memory.
- the stimuli may have a relation such that the subject may determine if the pairing or other association is correct or incorrect (e.g., are the stimuli paired such that the intended identifiable association between one element of a pair and the other element in the pair is present). For example, if a face - name pairing has been taught to the subject, the subject may be presented pairs of stimuli that have been scrambled such that the correct face - name pairing has been disrupted. The subject could then communicate whether the face - name pairing presented to it is the correct face - name pairing it has been taught.
- the pairs of stimuli may be presented in any suitable fashion.
- the pairs may be presented in random, organized, or a pseudo-random order during each testing session.
- the subjects may be tested over any suitable period at any suitable frequency.
- the subjects may be tested 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 50, 60, 75, 100, 150, 200, 250, 365, or any other suitable number of times per testing event (e.g., if testing is daily, the subject may be tested once, twice, three times, etc).
- subjects may be shown any suitable number of stimuli or pairs of stimuli.
- Subjects may have tests daily, every other day, weekly, bi-weekly, monthly, or any other suitable frequency. Moreover any suitable number of subjects may be tested. In some embodiments, testing is done in an effort to teach or encode the subject with the information as to the relationship between the paired stimuli. For example, the testing may be done to teach the subject which of the elements of the pair is the positive and/or negative element. In other embodiments, the testing may be done to teach the subject the correct pairing of elements within a set of stimuli (e.g., the correct face - name, occupation-name, or occupation-face/name association). In such embodiments, a plurality of phases of testing would likely be required. A first teaching phase, followed by a second phase wherein the subject's knowledge is tested by asking, through an appropriate method depending on the subject type (e.g., human vs. rat), if the presented pairings are correct.
- the subject type e.g., human vs. rat
- the subjects may be evaluated in any suitable manner.
- the subjects are presented with one or more pairs of stimuli in which one element of the pair is designated positive and/or one element of the pair is designated negative.
- the subject having learned which element of the pair is the positive and/or negative element, is evaluated on the number of times it selects the positive and/or negative element and/or does not select the positive and/or negative element.
- the pairs of stimuli may have a relation to one another (e.g., a face - name relation, occupation-name relation, occupation- face/name relation, a pair of words, a set of letters or numbers, parts of a picture or symbol).
- the subject may be evaluated on its ability, having learned the correct relation between the elements of the pair, to re-establish the appropriate pairs (e.g., placing the name element with the corresponding picture element, or the like).
- the pairs may be altered in some manner and the subject could be asked to identify the manner in which the pair was altered (for example, the response could be "intact,” “rearranged,” “one new,” or “both new”).
- a subject's long-term memory may be tested.
- a criterion result can be established. Such a criterion may be established in a variety of manners.
- the subject may be presented with the pair (or sets of pairs) until the subject identifies the positive and/or negative element with a certain frequency. That frequency would then serve as a criterion against which later results may be evaluated.
- the frequency of the correct establishment of the relation between elements of a pair of stimuli may also be used to establish a criterion.
- a subject is evaluated until it reaches a certain success rate. For example, a criterion of about 50%, 55%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99%,
- a subject may be tested a certain number of times or over a certain period of time, and the last evaluation may be used to establish the criterion. For example, a period of about 1-23 hours or about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 50, 60, 75, 100, 150, 200, 250, or 365 days, or any other suitable number of hours or days may be used to establish the criterion.
- the subject may be tested about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 50, 60, 75, 100, 150, 200, 250, 365, or any other suitable number of times to establish a criterion.
- the criterion may also be established using a mean, median, or other statistical measure of the subject's actual results.
- the subject is not evaluated or shown any of the pairs of stimuli used to establish the criterion for a certain amount of time.
- This amount of time may be about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 50, 60, 75, 100, 150, 200, 250, 365, or any other suitable number of days.
- the subject is once again tested using the same pairs of stimuli used to establish the criterion. In some embodiments, these tests are repeated under circumstances identical, similar, or different to those used in establishing the criterion.
- the subject's performance may be evaluated by establishing a long term success measurement in any suitable manner.
- the manner of establishing the long term success measurement may be similar to that described with respect to establishing a criterion, albeit using a generally shorter time period such that the long term success measurement is indicative of the subject's memory, rather than it having relearned the relevant criteria.
- the subject could then be evaluated by comparing the long term success measurement with the criterion. Statistical or similar analysis may be used to evaluate such results.
- the methods of the present invention may be used to evaluate the efficacy of a potential memory agent and/or a potential training protocol.
- the methods of the present invention may be used to identify a reduction in the amount of training required to reach a chosen performance level.
- a subject may first be evaluated in the absence of a memory agent and subsequently evaluated having been administered a memory agent.
- a control group of subjects could be tested using certain pairs of stimuli and those results compared to a group of subjects having been administered a given dose of a memory agent.
- the evaluation of the memory agent and/or training protocol may take a variety of forms. For example, a memory agent may reduce the time and/or number of days of training sessions required to establish or meet a certain criterion, or any other suitable measure of efficacy.
- the memory agent is evaluated to see if it alters (e.g., reduces or increases) the number of days or training sessions to reach a criterion of performance.
- criterion refers to a metric of a desired result and/or a desired level of performance such as, for example, a percentage of correct responses (e.g., correctly paired stimuli) from a test subject or group of test subjects. The criterion may be changed or chosen as desired by the administrator of the protocol, tests, tasks, and/or other embodiments of the present invention.
- the complexity of the training administered in these embodiments may be manipulated by, for example, changing the criterion, the numbers and/or types stimuli to be paired, the number of days of training, the quantity of training, and/or the type of training.
- the training, with or without the memory agent is sufficient to produce a CREB-dependent change in long term memory, for example a CREB-dependent enhancement of long term memory.
- the methods of the present invention may be used to evaluate the efficacy of a memory impairer. Such embodiments may involve steps similar to or identical to the methods for evaluating a memory agent. However, in such an embodiment, the evaluated indicia would be different.
- the memory impairer may increase the time required to establish a certain criterion, increase the number of training sessions required to reach a criterion, decrease the ratio of the long term success measurement to the criterion, or any may be evaluated in any other suitable manner.
- Any suitable memory agent or memory impairer may be provided by any suitable method, via any suitable delivery system and using any suitable delivery schedule and dose.
- any suitable training protocol may be used. Exemplary training protocols, memory agents and memory impairers are specifically discussed in the following sections.
- the present invention also includes systems, including systems for use along with the methods disclosed herein. For example, in some embodiments, the system may include all elements necessary to perform an embodiment of the methods of the present invention.
- Systems of the present invention may be useful for selecting a memory agent as a drug candidate, assessing the effectiveness of a memory agent, assessing the effectiveness of a training protocol, testing a memory agent as a long term memory enhancer, testing a memory agent as a impairer of specific or general memories, and/or testing a memory agent along with a training protocol as a long term memory enhancer.
- Such systems may involve use of sets of stimuli as described above, including paired stimuli, and testing schedules and protocols as described above.
- assessment using the methods and systems of the present invention can comprise two parts: (1) a specific training protocol and (2) administration of a memory agent.
- a specific training protocol In some embodiments, only the training protocol is used. In other embodiments, both training and administration of a memory agent occur.
- This combination can augment training protocols by reducing the number of days of training sessions required to yield a performance gain relative to that obtained with training alone or by requiring shorter or no rest intervals between training sessions to yield a performance gain.
- This combination can also augment training protocols by reducing the duration and/or number of training sessions required for the induction in a specific neuronal circuit(s) of a pattern of neuronal activity or by reducing the duration and/or number of training sessions or underlying pattern of neuronal activity required to induce cyclic AMP response element binding protein (CREB) - dependent long-term structural/function (i.e., long-lasting) change among synaptic connections of the neuronal circuit.
- CREB cyclic AMP response element binding protein
- Training protocols e.g., massed training, spaced training
- Administration of a memory agent in conjunction with training reduces the time and/or number of training sessions required to yield a gain in performance. As a result, less practice (training sessions) is required to learn the new language or to learn to play the new musical instrument.
- Training protocols are employed for repeated stimulation of neuronal activity or a pattern of neuronal activity underlying (a) specific neuronal circuit(s) in individuals.
- Administration of a memory agent in conjunction with training reduces the time and/or number of training sessions and/or underlying pattern of neuronal activity required to achieve a given result, such as induction of CREB-dependent long-term structure/function (i.e., long-lasting) change among synaptic connections of the neuronal circuit.
- these methods can enhance specific aspects of memory in an animal (particularly a human or other mammal or vertebrate) in need thereof by (a) administering to the animal a memory agent, for example one that enhances CREB pathway function; and (b) training the animal under conditions sufficient to produce an improvement in performance of a memory task of interest by the animal.
- the memory agent is a CREB pathway- enhancing drug.
- the described methods can be used to treat a memory deficit associated with a central nervous system (CNS) disorder or condition in an animal in need of said treatment by (a) administering to the animal a memory agent, for example one that enhances CREB pathway function; and (b) training the animal under conditions sufficient to produce an improvement in performance of a particular memory task by the animal.
- a memory agent for example one that enhances CREB pathway function
- CNS disorders and conditions include age-associated memory impairment, neurodegenerative diseases (e.g., Alzheimer's disease, Parkinson's disease, Huntington's disease (chorea), other senile dementia), psychiatric diseases (e.g., depression, schizophrenia, autism, attention deficit disorder), trauma dependent loss of function (e.g., cerebrovascular diseases (e.g., stroke, ischemia), brain tumor, head or brain injury), genetic defects (e.g., Rubinstein-Taybi syndrome, down syndrome), learning disabilities, and/or combinations thereof.
- neurodegenerative diseases e.g., Alzheimer's disease, Parkinson's disease, Huntington's disease (chorea), other senile dementia
- psychiatric diseases e.g., depression, schizophrenia, autism, attention deficit disorder
- trauma dependent loss of function e.g., cerebrovascular diseases (e.g., stroke, ischemia), brain tumor, head or brain injury
- genetic defects e.g., Rubinstein-Taybi syndrome, down
- the described methods can also be used for repeated stimulation of neuronal activity or a pattern of neuronal activity, such as that underlying a specific neuronal circuit(s), in an animal comprising (a) administering to the animal a memory agent, for example one that enhances CREB pathway function; and (b) training the animal under conditions sufficient to stimulate or induce neuronal activity or a pattern of neuronal activity in the animal.
- a memory agent for example one that enhances CREB pathway function
- spaced training produces a long- lasting memory that persists for at least seven days, is protein synthesis-dependent, is disrupted by over-expression of a CREB-repressor transgene and is normal in radish mutants (Tully, T. et al, Cell, 79(l):35-47 (1994); and Yin, J. C. et al, Cell, 79(l):49-58 (1994)).
- memory retention is composed of both the protein synthesis- and CREB- independent early memory (ARM) and the protein synthesis- and CREB-dependent long-term memory (LTM).
- mice LTM formation for both implicit and explicit tasks is defective in CREB mutant mice (Bourtchuladze, R. et al., Cell, 79(l):59-68 (1994)).
- a CREB-activator transgene abrogates the requirements for multiple, spaced training sessions and, instead, induces LTM formation after only one training session (which normally produces little or no memory retention 24 hours later (Yin, J. C. et al, Cell, 81(1): 107-115 (1995)).
- injection of a virally expressed CREB-activator transgene into rat amygdala also is sufficient to enhance memory after massed training for the fear-potentiated startle response, which abrogates the requirement for a rest interval in spaced training (Josselyn, S. A. et al., Society for Neuroscience, Vol. 24, Abstract 365.10 (1998)).
- LTM formation in CREB -deficient mice can form normally, if mutant mice are subjected to a different, spaced training protocol (Kogan, J. H. et al., Curr. Biol, 7(1): 1-11 (1997)).
- CREB also appears involved in various forms of developmental and cellular plasticity in the vertebrate brain. For example, neuronal activity increases CREB activity in the cortex (Moore, A. N. et al., J. Biol. Chem., 271(24): 14214- 14220 (1996)). CREB also mediates developmental plasticity in the hippocampus (Murphy, D. D. et al., Proc. Natl. Acad. Sci. USA, 94(4): 1482-1487 (1997)), in the somatosensory cortex (Glazewski, S. et al., Cereb. Cortex, 9(3):249-256 (1999)), in the striatum (Liu, F. C. et al., Neuron, 17(6): 1133-1144 (1996)), and in the visual cortex (Pham, T. A. et al., Neuron, 22(l):63-72 (1999)).
- CREB appears to be affected in human neurodegenerative disease and brain injury.
- CREB activation and/or expression is disrupted in Alzheimer's disease (Ikezu, T. et al., EMBO J., 15(10):2468-2475 (1996); Sato, N. et al., Biochem. Biophys. Res. Commun, 232(3):637-642 (1997); and Yamamoto-Sasaki, M. et al., Brain. Res., 824(2):300-303 (1999).
- CREB activation and/or expression is also elevated after seizures or ischemia (Blendy, J. A. et al, Brain Res., 681(l-2):8-14 (1995); and Tanaka, K.
- CREB functions during drug sensitivity and withdrawal.
- CREB is affected by ethanol (Pandey, S. C. et al., Alcohol Clin. Exp. Res., 23(9):1425-1434 (1999);
- a signal transduction pathway that can stimulate the CREB/CRE transcriptional pathway is the cAMP regulatory system. Consistent with this, mice lacking both adenylate cyclase 1 (ACl) and AC8 enzymes fail to learn (Wong S. T. et al., Neuron, 23(4):787-798 (1999)). In these mice, administration of forskolin to area CAl of the hippocampus restores learning and memory of hippocampal-dependent tasks. Furthermore, treatment of aged rats with drugs that elevate cAMP levels (such as rolipram and Dl receptor agonists) ameliorates an age-dependent loss of hippocampal-dependent memory and cellular long-term potentiation (Barad, M. et al., Proc.
- drugs that elevate cAMP levels such as rolipram and Dl receptor agonists
- ACT can be used to exercise appropriate neuronal circuits to fine-tune the synaptic connections of newly acquired, transplanted stem cells that differentiate into neurons.
- exercise appropriate neuronal circuit(s) is meant the induction in the appropriate neuronal circuit(s) of a pattern of neuronal activity, which corresponds to that produced by a particular cognitive training protocol.
- the cognitive training protocol can be used to induce such neuronal activity.
- neuronal activity can be induced by direct electrical stimulation of the neuronal circuitry.
- Neuronal activity and “neural activity” are used interchangeably herein.
- Memory agents may enhance CREB pathway function, which is required to consolidate newly acquired information into LTM.
- enhance CREB pathway function is meant the ability to enhance or improve CREB-dependent gene expression.
- CREB-dependent gene expression can be enhanced or improved by increasing endogenous CREB production, for example by directly or indirectly stimulating the endogenous gene to produce increased amounts of CREB, or by increasing functional (biologically active) CREB. See, e.g., U.S. Pat. No. 5,929,223; U.S. Pat. No. 6,051,559; and International Publication No. WO9611270 (published Apr. 18, 1996), which references are incorporated herein in their entirety by reference.
- Administration of a memory agent may decrease the training needed to yield a performance gain relative to that yielded with training alone.
- performance gain is meant an improvement in an aspect of memory performance.
- the invention provides methods for evaluating, testing and/or identifying the enhancement of a specific aspect of memory in an animal (particularly in a human or other mammal or vertebrate) (in some embodiments, in need thereof) wherein the enhancement may be due to the (a) administering to the animal a memory agent, for example one that enhances CREB pathway function; and (b) training the animal under conditions sufficient to produce an improvement in performance of a particular memory task by the animal.
- a memory agent for example one that enhances CREB pathway function
- Training can comprise one or multiple training sessions and is training appropriate to produce an improvement in performance of the memory task of interest. For example, if an improvement in language acquisition is desired, training would focus on language acquisition. If an improvement in ability to learn to play a musical instrument is desired, training would focus on learning to play the musical instrument. If an improvement in a particular motor skill is desired, training would focus on acquisition of the particular motor skill. The specific memory task of interest is matched with appropriate training.
- the invention also provides methods for evaluating, testing and/or identifying the enhancement of memory performance in an animal caused by repeated stimulation of neuronal activity or a pattern of neuronal activity, such as that underlying a specific neuronal circuit(s), in an animal comprising (a) administering to the animal a memory agent which enhances CREB pathway function; and (b) training the animal under conditions sufficient to stimulate or induce neuronal activity or a pattern of neuronal activity in the animal.
- training refers to training appropriate to stimulate or induce neuronal activity or a pattern of neuronal activity in the animal.
- multiple training sessions two or more training sessions.
- the memory agent can be administered before, during or after one or more of the training sessions. In a particular embodiment, the memory agent is administered before and during each training session.
- the invention also relates to methods of enhancing long term memory in an animal (particularly in a human or other mammal or vertebrate) in need of said treatment comprising (a) administering to the animal a memory agent identified, tested or evaluated by the methods of assessing, evaluating and/or testing the effectiveness of a memory agent, selecting a memory agent as a drug compound, or testing a memory agent as a long term memory enhancer, and (b) training the animal under conditions sufficient to enhance long term memory in the animal.
- the invention also relates to a method of treating an age-associated memory impairment in an animal in need of said treatment comprising (a) administering to the animal a memory agent identified, tested or evaluated by the methods of assessing, evaluating and/or testing the effectiveness of a memory agent, selecting a memory agent as a drug compound, or testing a memory agent as a long term memory enhancer; and (b) training the animal under conditions sufficient to enhance long term memory in the animal.
- the invention in another embodiment, relates to a method of treating a memory deficit associated with a neurodegenerative disease (e.g., Alzheimer's disease, Parkinson's disease, Huntington's disease, other senile dementia) in an animal in need of said treatment comprising (a) administering to the animal a memory agent identified, tested or evaluated by the methods of assessing, evaluating and/or testing the effectiveness of a memory agent, selecting a memory agent as a drug compound, or testing a memory agent as a long term memory enhancer; and (b) training the animal under conditions sufficient to enhance long term memory in the animal. .
- a neurodegenerative disease e.g., Alzheimer's disease, Parkinson's disease, Huntington's disease, other senile dementia
- the invention in another embodiment, relates to a method of treating a memory deficit associated with a psychiatric disease (e.g., depression, schizophrenia, autism, attention deficit disorder) in an animal in need of said treatment comprising (a) administering to the animal a memory agent identified, tested or evaluated by the methods of assessing, evaluating and/or testing the effectiveness of a memory agent, selecting a memory agent as a drug compound, or testing a memory agent as a long term memory enhancer; and (b) training the animal under conditions sufficient to enhance long term memory in the animal.
- a psychiatric disease e.g., depression, schizophrenia, autism, attention deficit disorder
- the invention in another embodiment, relates to a method of treating a memory deficit associated with trauma dependent loss of memory function (e.g., cerebrovascular diseases (e.g., stroke, ischemia), brain tumor, head or brain injury) in an animal in need of said treatment comprising (a) administering to the animal a memory agent identified, tested or evaluated by the methods of assessing, evaluating and/or testing the effectiveness of a memory agent, selecting a memory agent as a drug compound, or testing a memory agent as a long term memory enhancer; and (b) training the animal under conditions sufficient to enhance long term memory in the animal.
- trauma dependent loss of memory function e.g., cerebrovascular diseases (e.g., stroke, ischemia), brain tumor, head or brain injury
- the invention in another embodiment, relates to a method of treating a memory deficit associated with a genetic defect (e.g., Rubinstein-Taybi syndrome, down syndrome) in an animal in need of said treatment comprising (a) administering to the animal a memory agent identified, tested or evaluated by the methods of assessing, evaluating and/or testing the effectiveness of a memory agent, selecting a memory agent as a drug compound, or testing a memory agent as a long term memory enhancer; and (b) training the animal under conditions sufficient to enhance long term memory in the animal.
- a genetic defect e.g., Rubinstein-Taybi syndrome, down syndrome
- the invention also relates to methods of therapy of a memory deficit associated with a CNS disorder or condition in an animal having undergone neuronal stem cell manipulation (a) administering to the animal a memory agent identified, tested or evaluated by the methods of assessing, evaluating and/or testing the effectiveness of a memory agent, selecting a memory agent as a drug compound, or testing a memory agent as a long term memory enhancer; and (b) training the animal under conditions sufficient to enhance long term memory in the animal.
- neuronal stem cell manipulation is meant that (1) exogenous neuronal stem cells are transplanted into the brain or spinal chord of an animal or (2) endogenous neuronal stem cells are stimulated or induced to proliferate in the animal.
- Methods of transplanting neuronal stem cells into the brain or spinal chord of an animal are known and readily available in the art (see, e.g., Cameron, H. A. and McKay, R. D., Nat. Neurosci., 2:894-897 (1999); Kurimoto, Y. et al, Neurosci. Lett., 306:57-60 (2001); and Singh, G., Neuropathology, 21 :110-114 (2001)).
- Methods of stimulating or inducing proliferation of endogenous neuronal stem cells in an animal are known and readily available in the art (see, e.g., Gould, E. et al., Trends Cogn. Sci, 3:186- 192 (1999); Gould, E.
- the invention further relates to methods of improving or enhancing memory and/or performance in an animal with a learning, language or reading disability, or combinations of any of the foregoing, comprising (a) administering to the animal a memory agent identified, tested or evaluated by the methods of assessing, evaluating and/or testing the effectiveness of a memory agent, selecting a memory agent as a drug compound, or testing a memory agent as a long term memory enhancer; and (b) training the animal under conditions sufficient to enhance long term memory in the animal.
- Memory agents are compounds with pharmacological activity relative to memory disorders and/or memory dysfunctions, and include drugs, chemical compounds, ionic compounds, organic compounds, organic ligands, including cofactors, saccharides, recombinant and synthetic peptides, proteins, peptoids, nucleic acid sequences, including genes, nucleic acid products, and other molecules and compositions.
- Memory agents may, for example, enhance or impair short or long term memory.
- memory agents may be any of the compounds set forth in United States Patent Application Serial No. 11/679,782 entitled "Therapeutic Piperazones" and filed on February 27, 2007; United States Patent Application Serial No.
- a memory impairer are compounds with pharmacological activity relating to retained memories (for example, post-traumatic stress disorder), and include drugs, chemical compounds, ionic compounds, organic compounds, organic ligands, including cofactors, saccharides, recombinant and synthetic peptides, proteins, peptoids, nucleic acid sequences, including genes, nucleic acid products, and other molecules and compositions.
- the memory impairers and memory enhancers of the present invention can be used with any of the present methods and systems.
- memory agents can be cell permeant cAMP analogs (e.g., 8-bromo cAMP); activators of adenylate cyclase 1 (ACl) (e.g., forskolin); agents affecting G-protein linked receptor, such as, but not limited to adrenergic receptors and opioid receptors and their ligands (e.g., phenethylamines); modulators of intracellular calcium concentration (e.g., thapsigargin, N- methyl-D-aspartate (NMDA) receptor agonists); inhibitors of the phosphodiesterases responsible for cAMP breakdown (e.g., rolipram (which inhibits phosphodiesterase 4), iso-buto-metho- xanthine (IBMX) (which inhibits phosphodiesterases 1 and 2)); modulators of protein kinases and/or protein phosphatases, which mediate CREB protein activation and CREB-dependent gene
- Memory agents can also be exogenous CREB, CREB analogs, CREB-like molecules, biologically active CREB fragments, CREB fusion proteins, and/or nucleic acid sequences encoding exogenous CREB, CREB analogs, CREB-like molecules, biologically active CREB fragments and/or CREB fusion proteins.
- Memory agents can also be CREB function modulators, and/or nucleic acid sequences encoding CREB function modulators.
- CREB function modulators as used herein, have the ability to modulate CREB pathway function.
- modulate is meant the ability to change (increase or decrease) or alter CREB pathway function.
- Memory agents can be compounds which are capable of enhancing CREB function in the CNS. Such compounds include, but are not limited to, compounds which affect membrane stability and fluidity and specific immunostimulation. In a particular embodiment, the memory agent is capable of transiently enhancing CREB pathway function in the CNS.
- CREB analogs or derivatives, are defined herein as proteins having amino acid sequences analogous to endogenous CREB.
- Analogous amino acid sequences are defined herein to mean amino acid sequences with sufficient identity of amino acid sequence of endogenous CREB to possess the biological activity of endogenous CREB, but with one or more "silent" changes in the amino acid sequence.
- CREB analogs include mammalian CREM, mammalian ATF-I and other CREB/CREM/ATF-1 subfamily members.
- CREB-like molecule refers to a protein which functionally resembles (mimics) CREB. CREB-like molecules need not have amino acid sequences analogous to endogenous CREB.
- Examples of memory agents can include:
- Examples of memory agents can also include:
- Examples may also include a compound having the structure:
- each of Yi, Y 2 , Y 3 , and Y 4 is independently — H; straight chained or branched C 1 - C 7 alkyl, monofluoroalkyl or polyfluoroalkyl; straight chained or branched C 2 -C 7 alkenyl or alkynyl; C 3 -C 7 cycloalkyl, or C 5 -C 7 cycloalkenyl; --F, -Cl, -Br, or -I; -NO 2 ; -N 3 ; -CN; - OR 4 , -OCOR 4 , -COR 4 , -NCOR 4 , -N(R 4 ) 2 , -CON(R 4 ) 2 , or -COOR 4 ; aryl or heteroaryl; or any two of Yi, Y 2 , Y 3 and Y 4 present on adjacent carbon atoms can constitute a methylenedioxy group; wherein each R 4 is independently
- Ri and R 2 are each independently H, straight chained or branched Ci-C 7 alkyl, - F, --Cl, --Br, —I, -NO 2 , or --CN; wherein R 3 is H, straight chained or branched Ci-C 7 alkyl, -F, --Cl, --Br, —I, -NO 2 , --CN, -OR 6 , aryl or heteroaryl; wherein R5 is straight chained or branched Ci-C 7 alkyl, -N(R 4 ) 2 , — OR 4 or aryl; wherein R 6 is straight chained or branched Ci-C 7 alkyl or aryl; wherein B is C 3 -C 7 cycloalkyl, Cs-C 7 cycloalkenyl, adamantyl, aryl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazin
- A is aryl, heteroaryl, or heteroaryl (Ci-Ce)alkyl; and wherein aryl is substituted with — F, -Cl, --Br, —I, -NO 2 , --CN, straight chained or branched C 1 -C 7 alkyl, straight chained or branched C 1 -C 7 monofluoroalkyl, straight chained or branched Ci-C 7 polyfluoroalkyl, straight chained or branched C 2 -C 7 alkenyl, straight chained or branched C 2 -C 7 alkynyl, C 3 -C 7 cycloalkyl, C 3 -C 7 monofluorocycloalkyl, C 3 -C 7 polyfluorocycloalkyl, Cs-C 7 cycloalkenyl, -N(R-O 2 , -OR 4 , -SR 4 , -OCOR 4 , -COR
- A may also be aryl, heteroaryl, heteroaryl(Ci-C 6 )alkyl or ⁇ (CH 2 ) n ⁇ CC ⁇ R 4 ; wherein the aryl is substituted with —OH.
- A is A' and A' is
- each of Yi, Y 2 , Y 3 , and Y 4 is independently — H; straight chained or branched Ci-C 7 alkyl, --CF 3 , ⁇ F,-C1, -Br, -I, -OR 4 , -N(R-O 2 , or ⁇ CON(R 4 ) 2 ; wherein each R 4 is independently — H; straight chained or branched Ci-C 7 alkyl, -CF 3 , or phenyl; wherein A is A', straight chained or branched Ci-C 7 alkyl, aryl, heteroaryl, aryl(Ci-C 6 )alkyl or heteroaryl(Ci- Ce)alkyl; and wherein A' is
- B is C 3 -C 7 cycloalkyl or adamantyl.
- B is pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, indolizinyl, indol-4-yl, indol-5-yl, indol- 6- yl, indol-7-yl, isoindolyl, benzo[b]furan-4-yl, benzo[b]furan-5-yl, benzo[b]furan-6-yl, benzo[b]furan-7-yl, benzo[b]thiophen-4-yl, benzo[b]thiophen-5-yl, benzo[b]thiophen-6-yl, benzo[b]thiophen-7-yl, indazolyl, benzimidazolyl, benzo[b]thiazolyl, purinyl, imidazo[[b]thiazolyl
- B is aryl or is phenyl and the phenyl is optionally substituted with one or more of the following: — F, -Cl, --Br, -CF 3 , straight chained or branched Ci-C 7 alkyl, --OR 4 , -COR 4 , -NCOR 4 , -CO 2 R 4 , or -CON(R 4 ) 2 .
- each of Yi, Y 2 , Y3, and Y 4 is independently — H; straight chained or branched Ci- C 7 alkyl, monofluoroalkyl or polyfluoroalkyl; straight chained or branched C 2 -C 7 alkenyl or alkynyl; C 3 -C 7 cycloalkyl or C 5 -C 7 cycloalkenyl; -F, -Cl, -Br, or -I; -NO 2 ; -N 3 ; -CN; -OR 4 , -SR 4 , -OCOR 4 , -COR 4 , -NCOR 4 , -N(R-O 2 , -CON(R-O 2 , or -COOR 4 ; aryl or heteroaryl; or any two of Yi, Y 2 , Y 3 and Y 4 present on adjacent carbon atoms can constitute a methylenedioxy group; wherein each R 4 is independently
- Ri and R 2 are each independently H, straight chained or branched C 1 -C 7 alkyl, -
- R 3 is H, straight chained or branched Ci-C 7 alkyl, — F, --Cl, --Br, —I, --NO 2 , --CN, -OR 6 , aryl or heteroaryl; wherein R5 is straight chained or branched Ci-C 7 alkyl, -N(R-I) 2 , --OR 4 or aryl; wherein R 6 is straight chained or branched Ci-C 7 alkyl or aryl; wherein B is aryl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, indolizinyl, indol- 4-yl, indol-5-yl, indol-6-yl, indol-7-yl, isoindolyl, benzo[b]furan-4-yl
- each of Yi, Y 2 , Y 3 , and Y 4 is independently -H; straight chained or branched Ci-C 7 alkyl, -CF 3 , ⁇ F, --Cl, --Br, —I, ⁇ -OR 4 , -N(R-O 2 , or -CON(R 4 ) 2 .
- A is aryl, heteroaryl, heteroaryl(Ci-C 6 )alkyl or --(CH 2 ) J1 -CC-R 4 ; wherein the aryl is substituted with -OH.
- A is aryl, heteroaryl, or heteroaryl(Ci-C 6 )alkyl; and wherein aryl is substituted with — F, -Cl, -Br, —I, -NO 2 , -CN, straight chained or branched Ci-C 7 alkyl, straight chained or branched Ci-C 7 monofluoroalkyl, straight chained or branched Ci-C 7 polyfluoroalkyl, straight chained or branched C 2 -C 7 alkenyl, straight chained or branched C 2 -C 7 alkynyl, C 3 -C 7 cycloalkyl, C 3 -C 7 monofluorocycloalkyl, C 3 -C 7 monofluor
- A may also be aryl or aryl(Ci_C 6 )alkyl.
- A is A', straight chained or branched Ci-C 7 alkyl, aryl, heteroaryl, aryl(Ci- Ce)alkyl or heteroaryl(Ci-C 6 )alkyl; and A' is
- the compounds described herein may be enantiomerically and/or diastereomerically pure.
- The may also be a pure Z imine isomer or a pure Z alkene isome, or a pure E imine isomer or a pure E alkene isomer.
- Additional examples include:
- Additional examples may include a compound of the following formula:
- Ri, R 2 , Rd, and R 4 are each independently selected from the group consisting of hydrogen, hydroxy, halo, cyano, -CONRaRb, -NR 3 Rb, hydroxy(Ci-C6)alkyl , aryl, heteroaryl, heterocycle, amino(Ci-C 6 )alkyl, (Ci-Ce)alkyl optionally substituted with up to 5 fluoro, and (Ci- Ce)alkoxy optionally substituted with up to 5 fluoro; each R a and R b are independently hydrogen , (Ci-Ce)alkyl, aryl, (Ci-C 6 )alkylOC(O)-, or arylOC(O)-, or R a and R b are taken together with the nitrogen to which they are attached to form a heterocycle group optionally substituted with one or more Rj; wherein the heterocycle group optionally include one or more groups selected from O (oxygen), S(O) 2 , and NRc
- R c and R r are each independently selected from the group consisting of hydrogen, (C 1 - C 6 )alkyl, aryl, -S(O),(d-C 6 )alkyl, -S(O),aryl, -CONRg(Ci-C 6 alkyl), (Ci-C 6 )alky IC(O)-, aryl C(O)-, (Ci-C 6 )alkylOC(O)-, and arylOC(O)-;
- R g is hydrogen or (Ci-Ce)alkyl
- R 5 and R 6 are each independently selected from the group consisting of hydrogen, (Ci- Ce)alkyl, (C 2 -Ce)alkenyl, (C 2 -Ce)alkynyl, and aryl, or R 5 and Re are taken together with the nitrogen to which they are attached to form a heterocycle group optionally substituted with one or more Ra; wherein the heterocycle group optionally include one or more groups selected from O (oxygen), S(O) 2 , and NR C ; R 7 is selected from the group consisting of hydrogen, hydroxy, halo, hydroxy(Ci- Ce)alkyl, (Ci-Ce)alkyl optionally substituted with up to 5 fluoro, and (Ci-Ce)alkoxy optionally substituted with up to 5 fluoro;
- Ar is aryl, or heteroaryl, each optionally substituted with one or more Rs; and each R 8 is independently hydrogen, halo, CF 3 , CF 2 H, hydroxy, cyano, nitro,( Ci-Ce)alkyl, hydroxy(Ci-Ce)alkyl, (Ci-Ce)alkoxy, -NR 3 Rb, aryl, heteroaryl or heterocycle.
- R 5 and R 6 may, for example, together with the nitrogen to which they are attached, form a piperidinyl, pyrrolidinyl, morpholinyl, or thiomorpholinyl ring in the compound of the above formula.
- X is N(R C ), O (oxygen), C(Rd) 2 , or S(O) 2 ;
- z is an integer selected from 0, 1, and 2;
- each R d is independently selected from the group consisting of hydrogen, halo, oxo, hydroxy, -C(O)NRaRb, -NR 3 Rb, hydroxy(Ci-Ce)alkyl, aryl, aryl(Ci-Ce)alkyl, (Ci-Ce)alkyl optionally substituted with up to 5 fluoro , and (Ci-Ce)alkoxy optionally substituted with up to 5 fluoro; and
- n 0, 1, or 2.
- R 2 is Methyl, fluro, or OMe.
- R3 is Methyl, fluro, or OMe.
- R 2 and R3 are fluoro.
- R 2 and R3 are Methyl.
- each Y is independently N or
- R5 and R 6 together with the nitrogen to which they are attached, form a piperidinyl, pyrrolidinyl, morpholinyl, or thiomorpholinyl ring, each optionally substituted with one or more Rd.
- n can be 0, 1 or 2.
- R 2 may be Methyl, fluoro or OMe.
- R 3 can be Methyl, fluoro or OMe.
- R 2 and R3 can be fluoro.
- R 2 and R3 can be Methyl.
- Examples also include a compound of the following formula:
- W is O, S, or two hydrogens;
- X is 0 or N-Y-R 4 ;
- R a is H, (Ci-C 6 )alkyl, (C 3 -C 8 )cycloalkyl, (Ci-C 6 )alkoxy(C 2 -C 6 )alkyl, or (C 3 - C 8 )cycloalkyl(Ci-C 6 )alkyl;
- Z is a phenyl ring substituted with one or more substituents independently selected from (Ci-C 6 )alkyl, halo(d-C 6 )alkyl, (C 3 -C 8 )cycloalkyl, (C 3 -C 8 )cycloalkyl(Ci-C 6 )alkyl, (C 1 - Ce)alkoxy, halo(Ci-Ce)alkoxy, (C 3 -C 8 )cycloalkyloxy, and (C 3 -C 8 )cycloalkyl(Ci-C6)alkoxy; or Z is a phenyl ring that is fused to a saturated, partially unsaturated, or aromatic, mono- or bicyclic ring system comprising from about 3 to about 8 atoms selected from carbon, oxygen, and NR b , wherein the mono- or bicyclic ring system of Z is optionally substituted with one or more Rb, and wherein the phenyl ring that is
- R b is absent, H, (Ci-C 6 )alkyl, (C 3 -C 8 )cycloalkyl, (Ci-C 6 )alkoxy(C 2 -C 6 )alkyl, or (C 3 - C 8 )cycloalkyl(Ci-C 6 )alkyl;
- Rc is (Ci-C 6 )alkyl, (Ci-C 6 )alkoxy, (Ci-C 6 )alkanoyl, (Ci-C 6 )alkoxycarbonyl, (C 1 -
- each R d and R e is independently H, hydroxy, (Ci-Ce)alkyl, (Ci-Ce)alkenyl, (C 2 - C 6 )alkynyl, (Ci-C 6 )alkoxy, (C 2 -C 6 )alkenyloxy, (C 2 -C 8 )alkyl, halo(C 1 -C 6 )alkyl, aryl, aryl(Ci-C 6 )alkyl, aryl(Ci-C 6 )alkoxy, aryl(C 1 -C 6 )alkanoyl, het, het(C 1 -C 6 )alkyl, het(Ci-C 6 )alkoxy, or het(Ci-C6)alkanoyl; each R d and R e is independently H, hydroxy, (Ci-Ce)alkyl, (Ci-Ce)alkenyl, (C 2
- each Rf and R g is independently H, (Ci-Ce)alkyl, (Ci-Ce)alkoxy, (Ci-Ce)alkanoyl, (C 1 - C 6 )alkoxycarbonyl, (C 3 -C 8 )cycloalkyl, (C 3 -C 8 )cycloalkyl(C 1 -C 6 )alkyl, aryl, aryl(d-C 6 )alkyl, or aryl(Ci-Ce)alkoxy; or Rf and R g together with the nitrogen to which they are attached form a pyrrolidin
- R 1 is (Ci-Ce)alkyl, (Ci -C 6 )haloalkyl, or aryl , unsubstituted or substituted with one or more R 6 ; one of R 2 and R 3 is absent and the other is hydrogen, (Ci-Ce)alkyl, halo(Ci-Ce)alkyl, hydroxy(Ci-Ce)alkyl, (C 3 -C 8 )cycloalkyl, amino(C 2 -C 6 )alkyl, or aryl, each unsubstituted or substituted with one or more groups selected from alkyl, halo, haloalkyl or nitro, Het, (C 3 - C 8 )cycloalkyl(Ci-C 6 )alkyl, aryl(d-C 6 )alkyl, (C 1 -C 6 ) or Het(C r C 6 )alkyl;
- B is aryl, thiophene, heteroaryl, furan or pyrrole
- Y is R 4 , -N(R 4 ) 2 , -OR 4 , -SR 4 , or -C(R 4 ) 3 ; each R 4 is independently selected from the group consisting of hydrogen, (CiC 6 ) alkyl, (C 2 -C 6 )alkenyl, (C 2 -C 6 )alkynyl, (Ci-C 6 )alkoxy, (Ci-C 6 )alkanoyl, (C 1 C 6 )alkoxycarbonyl, (C 3 - C 8 )cycloalkyl, (C 3 -C 8 )cycloalkyl(C 1 -C 6 )alkyl, (Ci-C 6 )alkoxy(C 2 -C 6 )alkyl, hydroxy(C 2 -C 6 )alkyl, cyano(Ci-C 6 )alkyl, (Ci-C 6 )alkthio(Cz-C 6 )alky
- R 1 is (Ci-Ce)alkyl, (Ci-C 6 )haloalkyl, or aryl, unsubstituted or substituted with one or more R 6 ; one of R 2 and R 3 is absent and the other is hydrogen, (Ci-Ce)alkyl, halo(Ci-Ce)alkyl, hydroxy(Ci-Ce)alkyl, (C 3 -Cg)cycloalkyl, amino(C 2 -C 6 )alkyl, or aryl, each unsubstituted or substituted with one or more groups selected from alkyl, halo, haloalkyl or nitro, Het, (C 3 - C 8 )cycloalkyl(Ci-C 6 )alkyl, aryl(d-C 6 )alkyl, (C 1 -C 6 ) or Het(C r C 6 )alkyl; each of Z 1 , Z 2 , and Z 3 is independently
- At least one of Z 1 or Z 2 must be N(R% O, or S; each p is independently 0, 1, or 2; each q is independently 0 or 1 ;
- each R e is independently selected from halo, hydroxy, cyano, nitro, azido, (Ci-C6)alkyl, Het, aryl, (C 1 -C 6 )alkylHet, (Ci-C 6 )alkylaryl, (C 1 -C 6 )alkylHet(C 1 -C 6 )alkyl, (Ci-C 6 )alkyla
- Biologically active polypeptide fragments of CREB can include only a part of the full- length amino acid sequence of CREB, yet possess biological activity. Such fragments can be produced by carboxyl or amino terminal deletions, as well as internal deletions.
- Fusion proteins comprise a CREB protein as described herein, referred to as a first moiety, linked to a second moiety not occurring in the CREB protein.
- the second moiety can be a single amino acid, peptide or polypeptide or other organic moiety, such as, without limitation, a carbohydrate, a lipid or an inorganic molecule.
- Nucleic acid sequences are defined herein as heteropolymers of nucleic acid molecules.
- the nucleic acid molecules can be double stranded or single stranded and can be a deoxyribonucleotide (DNA) molecule, such as cDNA or genomic DNA, or a ribonucleotide (RNA) molecule.
- the nucleic acid sequence can, for example, include one or more exons, with or without, as appropriate, introns, as well as one or more suitable control sequences.
- the nucleic acid molecule contains a single open reading frame which encodes a desired nucleic acid product.
- the nucleic acid sequence can be operably linked to a suitable promoter.
- a nucleic acid sequence encoding a desired CREB protein, CREB analog (including CREM, ATF-I), CREB-like molecule, biologically active CREB fragment, CREB fusion protein or CREB function modulator can be isolated from nature, modified from native sequences or manufactured de novo, as described in, for example, Ausubel et al., Current Protocols in Molecular Biology, John Wiley & Sons, New York (1998); and Sambrook et al., Molecular Cloning: A Laboratory Manual, 2nd edition, Cold Spring Harbor University Press, New York. (1989). Nucleic acids can be isolated and fused together by methods known in the art, such as exploiting and manufacturing compatible cloning or restriction sites.
- the nucleic acid sequence will be a gene which encodes the desired CREB protein, CREB analog, CREB-like molecule, CREB fusion protein or CREB function modulator.
- a gene is typically operably linked to suitable control sequences capable of effecting the expression of the CREB protein or CREB function modulator, preferably in the CNS.
- operably linked is defined to mean that the gene (or the nucleic acid sequence) is linked to control sequences in a manner which allows expression of the gene (or the nucleic acid sequence). Generally, but not always, operably linked means contiguous.
- Control sequences include a transcriptional promoter, an optional operator sequence to control transcription, a sequence encoding suitable messenger RNA (mRNA) ribosomal binding sites and sequences which control termination of transcription and translation.
- mRNA messenger RNA
- a recombinant gene or a nucleic acid sequence
- encoding a CREB protein, CREB analog, CREB-like molecule, biologically active CREB fragment, CREB fusion protein or CREB function modulator can be placed under the regulatory control of a promoter which can be induced or repressed, thereby offering a greater degree of control with respect to the level of the product.
- promoter refers to a sequence of DNA, usually upstream (5') of the coding region of a structural gene, which controls the expression of the coding region by providing recognition and binding sites for RNA polymerase and other factors which may be required for initiation of transcription. Suitable promoters are well known in the art. Exemplary promoters include the SV40 and human elongation factor (EFI).
- EFI human elongation factor
- Memory agents can enhance CREB pathway function by a variety of mechanisms.
- a memory agent can affect a signal transduction pathway which leads to induction of CREB -dependent gene expression.
- Induction of CREB-dependent gene expression can be achieved, for example, via up-regulation of positive effectors of CREB function and/or down- regulation of negative effectors of CREB function.
- Positive effectors of CREB function include adenylate cyclases and CREB activators.
- Negative effectors of CREB function include cAMP phosphodiesterase (cAMP PDE) and CREB repressors.
- a memory agent can enhance CREB pathway function by acting biochemically upstream of or directly acting on an activator or repressor form of a CREB protein and/or on a CREB protein containing transcription complex.
- CREB pathway function can be affected by increasing CREB protein levels transcriptionally, post-transcriptionally, or both transcriptionally and post-transcriptionally; by altering the affinity of CREB protein to other necessary components of the of the transcription complex, such as, for example, to CREB- binding protein (CBP protein); by altering the affinity of a CREB protein containing transcription complex for DNA CREB responsive elements in the promoter region; or by inducing either passive or active immunity to CREB protein isoforms.
- CBP protein CREB- binding protein
- Memory agents can be administered directly to an animal in a variety of ways.
- memory agents are administered systemically.
- Other routes of administration are generally known in the art and include intravenous including infusion and/or bolus injection, intracerebroventricularly, intrathecal, parenteral, mucosal, implant, intraperitoneal, oral, intradermal, transdermal (e.g., in slow release polymers), intramuscular, subcutaneous, topical, epidural, etc. routes.
- Other suitable routes of administration can also be used, for example, to achieve absorption through epithelial or mucocutaneous linings.
- Particular memory agents can also be administered by gene therapy, wherein a DNA molecule encoding a particular therapeutic protein or peptide is administered to the animal, e.g., via a vector, which causes the particular protein or peptide to be expressed and secreted at therapeutic levels in vivo.
- a vector refers to a nucleic acid vector, e.g., a DNA plasmid, virus or other suitable replicon (e.g., viral vector).
- Viral vectors include retrovirus, adenovirus, parvovirus (e.g., adeno-associated viruses), coronavirus, negative strand RNA viruses such as orthomyxovirus (e.g., influenza virus), rhabdovirus (e.g., rabies and vesicular stomatitis virus), paramyxovirus (e.g.
- RNA viruses such as picomavirus and alphavirus
- double stranded DNA viruses including adenovirus, herpesvirus (e.g., Herpes Simplex virus types 1 and 2, Epstein-Barr virus, cytomegalovirus), and poxvirus (e.g., vaccinia, fowlpox and canarypox).
- herpesvirus e.g., Herpes Simplex virus types 1 and 2, Epstein-Barr virus, cytomegalovirus
- poxvirus e.g., vaccinia, fowlpox and canarypox
- Other viruses include Norwalk virus, togavirus, flavivirus, reoviruses, papovavirus, hepadnavirus, and hepatitis virus, for example.
- retroviruses examples include: avian leukosis-sarcoma, mammalian C-type, B-type viruses, D-type viruses, HTLV-BLV group, lentivirus, spumavirus (Coffin, J. M., Retroviridae: The viruses and their replication, In Fundamental Virology, Third Edition, B. N. Fields, et al., Eds., Lippincott-Raven Publishers, Philadelphia, 1996).
- murine leukemia viruses include murine leukemia viruses, murine sarcoma viruses, mouse mammary tumor virus, bovine leukemia virus, feline leukemia virus, feline sarcoma virus, avian leukemia virus, human T-cell leukemia virus, baboon endogenous virus, Gibbon ape leukemia virus, Mason Pfizer monkey virus, simian immunodeficiency virus, simian sarcoma virus, Rous sarcoma virus and lentiviruses.
- vectors are described, for example, in McVey et al., U.S. Pat. No. 5,801,030, the teachings of which are incorporated herein by reference.
- a nucleic acid sequence encoding a protein or peptide e.g., CREB protein, CREB analog
- CREB-like molecule biologically active CREB fragment, CREB fusion protein, CREB function modulator
- methods generally known in the art see, e.g., Ausubel et al., Eds., Current Protocols in Molecular Biology, John Wiley & Sons, Inc., New York (1998); Sambrook et al., Eds., Molecular Cloning: A Laboratory Manual, 2nd edition, Cold Spring Harbor University Press, New York (1989)).
- the mode of administration is preferably at the location of the target cells. In a particular embodiment, the mode of administration is to neurons.
- Memory agents can be administered together with other components of biologically inactive agents, such as pharmaceutically acceptable surfactants or carriers (e.g., glycerides), excipients (e.g., lactose), stabilizers, preservatives, humectants, emollients, antioxidants, carriers, diluents and vehicles. If desired, certain sweetening, flavoring and/or coloring agents can also be added.
- a memory agent may also be referred to as a pharmaceutical composition.
- Memory agents can be formulated as a solution, suspension, emulsion or lyophilized powder in association with a pharmaceutically acceptable parenteral vehicle.
- a pharmaceutically acceptable parenteral vehicle examples include water, saline, Ringer's solution, isotonic sodium chloride solution, dextrose solution, and 5% human serum albumin. Liposomes and nonaqueous vehicles such as fixed oils can also be used.
- the vehicle or lyophilized powder can contain additives that maintain isotonicity (e.g., sodium chloride, mannitol) and chemical stability (e.g., buffers and preservatives).
- the formulation can be sterilized by commonly used techniques. Suitable pharmaceutical carriers are described in Remington's Pharmaceutical Sciences.
- the dosage of memory agent administered to an animal is that amount required to effect a change in CREB-dependent gene expression, particularly in neurons.
- the dosage administered to an animal, including frequency of administration will vary depending upon a variety of factors, including pharmacodynamic characteristics of the particular memory agent, mode and route of administration; size, age, sex, health, body weight and diet of the recipient; nature and extent of symptoms being treated or nature and extent of the memory being enhanced or modulated, kind of concurrent treatment, frequency of treatment, and the effect desired.
- Memory agents can be administered in single or divided doses (e.g., a series of doses separated by intervals of days, weeks or months), or in a sustained release form, depending upon factors such as nature and extent of symptoms, kind of concurrent treatment and the effect desired.
- Other therapeutic protocols or agents can be used in conjunction with the present invention.
- animals are presented with 2 pairs of stimuli (e.g. 1 pair of shapes and 1 pair of lines) over the course of a daily test session.
- one stimulus is arbitrarily designated as the positive stimulus (i.e., touch of that stimulus is rewarded).
- Each pair of stimuli is presented 14 times during a daily session that consists of 28 trials.
- the pairs of stimuli are presented in a pseudo randomized fashion during each session. Animals have to touch the "positive" shape or line in order to receive a sugar pellet reward accompanied by a pleasant tone. Touching the alternative shape or line results in a negative tone, no sugar pellet and a time out.
- Animals are tested daily until they reach a performance criterion of 85% correct responses during a daily session. When this criterion is reached, animals receive no further testing for the next 7 days. On day 8 post-criterion, animals are tested again using the same pairs of stimuli that they have seen previously learned in order to assess long term retrieval of the learned discriminations.
- the treatment schedule is shown in Table 1. Treatments were administered sequentially starting with treatment 1. Each treatment (dose of HT-0712 or vehicle) was administered once per day for 4 days before behavioral training began and then continued at 1 treatment (dose of HT-0712 or vehicle) per day, delivered four hours before testing, for the duration of the training period. When each individual animal reached performance criterion of at least 85% correct responses during a daily session, training and dosing ceased for seven days. Then, on post- criterion day 8, animals were tested again on the DL LTR task (retention testing).
- a second vehicle study was conducted at the end of the drug trials using the first 3 animals that completed the 100mg/kg dosing study.
- the percent correct responses on the total number of daily trials were recorded as well as the number of days needed to reach the criterion (at least 85% correct responses in a single test session).
- the percent correct responses on the retention trial was recorded for the entire retention trial session and sub-analyzed for performance differences on trials 1-14 versus trials 15-28.
- the second (post drug study) vehicle study was performed using the first 3 animals that finished the drug trials. During this second vehicle study, animals reached criterion over a period of 20.0 ⁇ 4.5 days. Their mean criterion value was 86.3 ⁇ 1.3% correct responses.
- Performance at the retention trials averaged 64.0 ⁇ 4.0% correct responses.
- retention trial performance was divided between trials 1 to 14 versus trials 15 to 28, animals performed 59.3 ⁇ 8.4% correct responses and 69.0 ⁇ 10.4% correct responses, respectively.
- DL LTR Discrimination Learning - Long Term Retrieval task
- the results of the vehicle control studies indicate that normal aged animals require a long time (ex. > 20 days) to reach an 85% correct response criterion on this test. Additionally, the post-drug vehicle test results indicate that the increase in performance seen with HT-0712 was not due to an increase in the animal's proficiency in the test or a practice effect as the second vehicle control test results were not significantly different from the results of the first vehicle study.
- the response of the animals could be separated into 2 general groups.
- the best dose for improving performance was the lmg/kg dose of HT- 0712.
- These animals showed a decrement in performance as the dose administered increased from 1 to 10 to 100mg/kg. That is, with increasing doses of HT-0712, it took longer for the animals to reach criterion (i.e., 4, 5, and 10 days to criterion (old-5) and 3, 12, and 18 days to criterion for (old-8) after administration of 1, 10, or 100 mg/kg, respectively).
- criterion i.e., 4, 5, and 10 days to criterion (old-5) and 3, 12, and 18 days to criterion for (old-8) after administration of 1, 10, or 100 mg/kg, respectively.
- the second group of animals (old-1, old-2 and old-7) showed a more typical dose-response effect in which performance improved with increasing doses (21, 15, and 3 days to criterion; 17, 18, and 11 days to criterion, and 15, 9, and 3 days to criterion for old-1, old2 and old-7 respectively with doses of 1, 10, and 100 mg/kg).
- the present animals were previously tested and assessed for cognitive deficits using both delayed matching to sample (DMTS) and paired associative learning (PAL) tasks. These animals were ranked accordingly to their level of performance on these tasks (i.e., animals were ranked per task from 1 to 6, 1 being the best performer and 6 the worst (see Table 10). This ranking showed that the animal that did not respond to HT-0712 was ranked 1 on performance of the PAL and 2 on performance of the DMTS. This same animal also had the best performance on the first vehicle study performed on the current project.
- DMTS delayed matching to sample
- PAL paired associative learning
- RT retention test
- RT 1-14 Retention test trials 1 to 14
- RT 15-28 retention test trials 15 to 28
- RT retention test
- RT 1-14 Retention test trials 1 to 14
- RT 15-28 retention test trials 15 to 28
- RT retention test
- RT 1-14 Retention test trials 1 to 14
- RT 15-28 retention test trials 15 to 28
- Table 10 Ranking of animals used in the study on PAL and DMTS
- PAL Mean number of trials needed to complete the most difficult level of the task (3 stimuli / 3 locations).
- DMTS Mean number of correct response at the longer delay.
- EXAMPLE 2 SINGLE-BLIND STUDY IN HEALTHY, ELDERLY, HUMAN SUBJECTS
- a long term retrieval task can utilize stimuli in the form of face - name and/or occupation pairings.
- the human subjects can be shown a picture of a face along with the name and/or occupation of a person. An occupation may be listed for each name-face pair.
- the human subject is asked to decide if the name fits with the occupation.
- the human subject is shown intact, rearranged, and novel face-name pairs (presented without the occupation) and asked to decide if the face and name pairings are intact, rearranged, or novel.
- human subjects can then be evaluated in a manner as set forth in Example 1.
- a single site, double-blind, three-way cross-over study was performed that tested the following three dosing regimens: (1) HT-0712 45 mg po for 7 days; (2) HT-0712 45 mg po plus warfarin 2.5 mg po for 7 days; and (3) Warfarin 2.5 mg po for 7 days. There was a seven-day washout periods between dosing periods. Subjects remained in- house for three (72 hours) of the seven days washout days, for study related procedures. Subjects were furloughed from the clinic for four of the seven washout days. With regard to subject selection criteria, there were no inclusion and/or exclusion criteria for the paired associate test. Study Schedule
- Group I subjects received 45 mg doses of HT-0712 on Days 1,2,3,4,5,6, and 7; 45 mg doses of HT-0712 plus 2.5 mg doses of warfarin on Days, 15, 16, 17, 18, 19, 20, and 21; and 2.5 mg doses of warfarin on Days 29, 30, 31, 32,33, 34, and 35.
- Group II subjects received 45 mg doses of HT-0712 plus 2.5 mg doses of warfarin on Days 1,2,3,4,5,6, and 7; 2.5 mg doses of warfarin on Days 15, 16, 17, 18, 19, 20, and 21; and 45 mg doses of HT-0712 on Days 29, 30, 31, 32,33, 34, and 35.
- Group III subjects received 2.5 mg doses of warfarin on Days 1, 2,3,4,5,6, and 7; 45 mg doses of HT-0712 on Days 15, 16, 17, 18, 19, 20, and 21; and 45 mg doses of HT-0712 plus 2.5 mg doses of warfarin on Days 29, 30, 31, 32, 33, 34, and 35.
- Paired Associates Testing Materials Materials consisted of 21 Lenovo notebook computers. Each notebook was configured to eliminate all auto-configuration software and reduce settings which might interfere with the internal clock settings and E-Run software. E-Run (vl .2.1.68) was installed on all the notebook computers to administer the paired associate test. 120 grayscale faces (male and female) were paired with a two-word professions, such as "fire fighter.” Grayscale face images were drawn from a face database from Lund University Cognitive Science Lab. Only two-word or compound- word professions were used. The 120 face-name pairs were divided into three groups of 40. During each round the same 40 face-name pairs were used throughout the round.
- Procedure - Timing of Administration Memory testing commenced approximately 6 hours post-drug administration. All memory testing was completed prior to daily blood samples being taken. The training was performed in three Rounds (or Segments) consisting of 14 days as shown in Table 12. The testing was performed over 42 days. In a given round on day 1, Subjects were familiarized with the computers and trained how to use the test software. On day 2 subjects were given an encoding and retrieval session. On days 3-7, subjects were tested daily for recall. On day 10 subjects were tested on half the face-name pairs, and on day 14, subjects were tested on the remaining half of the face-name pairs followed by a complete test of all face- name pairs used in that round.
- a group of items linked with the conjunction "and” should not be read as requiring that each and every one of those items be present in the grouping, but rather should be read as “and/or” unless expressly stated otherwise.
- a group of items linked with the conjunction "or” should not be read as requiring mutual exclusivity among that group, but rather should also be read as “and/or” unless expressly stated otherwise.
- items, elements or components of the invention may be described or claimed in the singular, the plural is contemplated to be within the scope thereof unless limitation to the singular is explicitly stated.
Abstract
Description
Claims
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IL210696A IL210696A (en) | 2008-07-18 | 2011-01-17 | Method for assessing the effectiveness of a memory agent administered to a primate in enhancing long term memory |
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Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20070179197A1 (en) * | 2000-05-01 | 2007-08-02 | Accera, Inc. | Compositions and methods for improving or preserving brain function |
US20080039496A1 (en) * | 2006-06-26 | 2008-02-14 | Blackburn Thomas P | Method of Modulating Neurite Outgrowth by the use of a Galanin-3 Receptor Antagonist |
US20080160534A1 (en) * | 2002-06-25 | 2008-07-03 | Tully Timothy P | Altering memory by affecting staufen |
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US8124589B2 (en) * | 2000-05-01 | 2012-02-28 | Accera, Inc. | Use of ketogenic compounds for treatment of age-associated memory impairment |
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JP2002020291A (en) * | 2000-06-30 | 2002-01-23 | Sumitomo Pharmaceut Co Ltd | Therapeutic agent for cognitive dysfunction |
US20040224316A1 (en) * | 2000-08-10 | 2004-11-11 | Tully Timothy P. | Augmented cognitive training |
ATE526020T1 (en) * | 2000-08-10 | 2011-10-15 | Cold Spring Harbor Lab | INCREASED COGNITIVE TRAINING |
US7642281B2 (en) * | 2002-08-07 | 2010-01-05 | Helicon Therapeutics, Inc. | Indolone compounds useful to treat cognitive impairment |
PT1540335E (en) * | 2002-08-19 | 2010-06-23 | Helicon Therapeutics Inc | Screening methods for cognitive enhancers |
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BRPI0711816A2 (en) * | 2006-05-19 | 2011-12-13 | Helicon Therapeutics Inc | phosphodiesterase 4 inhibitors for motor and cognitive rehabilitation |
GB0611458D0 (en) * | 2006-06-09 | 2006-07-19 | Univ Cambridge Tech | Assessment of functional status |
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Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20070179197A1 (en) * | 2000-05-01 | 2007-08-02 | Accera, Inc. | Compositions and methods for improving or preserving brain function |
US20080160534A1 (en) * | 2002-06-25 | 2008-07-03 | Tully Timothy P | Altering memory by affecting staufen |
US20080039496A1 (en) * | 2006-06-26 | 2008-02-14 | Blackburn Thomas P | Method of Modulating Neurite Outgrowth by the use of a Galanin-3 Receptor Antagonist |
Non-Patent Citations (1)
Title |
---|
See also references of EP2313098A4 * |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US8277842B1 (en) | 2012-01-20 | 2012-10-02 | Dart Neuroscience (Cayman) Ltd. | Enteric-coated HT-2157 compositions and methods of their use |
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BRPI0916228A2 (en) | 2015-11-03 |
CA2730940A1 (en) | 2010-01-21 |
IL210696A0 (en) | 2011-03-31 |
JP2011528667A (en) | 2011-11-24 |
EP2313098A4 (en) | 2011-12-21 |
JP5789511B2 (en) | 2015-10-07 |
SG195572A1 (en) | 2013-12-30 |
CN102143749A (en) | 2011-08-03 |
IL210696A (en) | 2014-12-31 |
AU2009270697A1 (en) | 2010-01-21 |
KR20110049801A (en) | 2011-05-12 |
MX2011000652A (en) | 2011-06-16 |
EP2313098A1 (en) | 2011-04-27 |
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