EP0260312A1 - Method for blocking calcium channels - Google Patents
Method for blocking calcium channelsInfo
- Publication number
- EP0260312A1 EP0260312A1 EP87902204A EP87902204A EP0260312A1 EP 0260312 A1 EP0260312 A1 EP 0260312A1 EP 87902204 A EP87902204 A EP 87902204A EP 87902204 A EP87902204 A EP 87902204A EP 0260312 A1 EP0260312 A1 EP 0260312A1
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- EP
- European Patent Office
- Prior art keywords
- alcohol
- cells
- mammal
- composition
- alcohols
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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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
- A61K31/045—Hydroxy compounds, e.g. alcohols; Salts thereof, e.g. alcoholates
Definitions
- This invention relates to methods and compositions for regulating calcium conductance across cell membranes. More particularly, this invention relates to methods and composi ⁇ tions for blocking "low-threshold" calcium chan ⁇ els and for treating the symptoms of tremors and other pathological condi ⁇ tions associated with, or controlled by, low-threshold calcium conductance.
- Neurons of the central nervous system transmit informa ⁇ tion via electrical impulses. These impulses are generated by electrochemical potentials caused by the movement of charged particles across (i.e. through) the cell membrane.
- the size of the impulse transmitted is a function of the membrane conduc ⁇ tance.
- the term "conductance,” as applied to transmission of ionic charge across a cell membrane, means the incremental cur ⁇ rent (or current-like) response exhibited through the cell mem- brane as a result of the application or a voltage (an incre ⁇ mental change in the electric field) across the membrane.
- a calcium channel is a structure that spans the thickness of the lipid bilayer in cell membranes and allows calcium ions to move passively across this lipid bilayer ac- cording to the calcium electrochemical gradient between the interior of the cell (where calcium concentration is lower) and the extra-cellular fluid (where calcium concentration is high er) .
- These channels demonstrate a higher specificity for the movement of the calcium ion across this channel-like structure, although a calcium channel's selectivity with respect to other divalent cations is never perfect.
- Calcium channels can be activated by changes in the electric field of the cell mem ⁇ brane.
- a calcium pump is a different structural moiety (proba ⁇ bly one or more macromolecules) of the cell membrane. This moiety spans the cell membrane and produces an active movement of calcium ions against their electrochemical gradient. Ion pumps are therefore different from ion channels in that they require energy to generate ionic movement since ionic flow through a pump occurs against the electrochemical gradient.
- Direct stimulation of neurons in vitro produces action potentials having two main components: a fast spike, due to sodium conductance, and a slower, calcium-dependent spike (Llinas, R. and Sugi ori, M. J__ Physiol. 305:197-213 1980).
- Sodium-dependent potentials are the prominant feature of the somatic (or cell-body) response, while calcium-dependent action potentials are more apparent in the dendrites.
- the ADP was followed by an after- hyperpolarization potential (AHP) , 3.
- AHP after- hyperpolarization potential
- the rebound spike (RS) was shown to be due to the activation of low- threshold calcium conductance (LTCC) .
- the shaded areas in Fig. 1 represent the two types of voltage spikes due to the two types of calcium conductance.
- the ADP and AHP are tetrodo- toxin-insensitive (textrodoxin inhibits sodium conductance) and, therefore, they cannot be due to sodium conductance.
- the following can be considered an operative definition of the RS (or LTC spike) :
- the RS is generated by the presence of LTCC as follows:
- the RS occurs as the membrane is abruptly depolarized with square voltage pulses of increasing amplitude, if the membrane potential is more negative than -70mV.
- the threshold in the inferior olivary cells studied by the present inventio is -65mV.
- the RS occurs as a rebound action potential fol lowing hyperpolarization of the cell membrane from a restin potential.
- the firing level of the RS spike for the inferio olivary cells studied by the present inventors is the same a in (a) , i.e. -65mV.
- the RS When a voltage clamp technique (described below) is used, the RS generates an ionic current that occurs at nega tive values of cell membrane potential, rises to a maximum, an is then inactivated — also within the negative potential rang (at about -45mV for the I.O. cells studied) .
- HTCC is involved in dendridic action potentials, i certain components of heart action, and in synaptic transmis sion. LTCC appears to be a somatic response.
- Tremor consists of a more or less regular rhythmic os cillation of a part of the body about a fixed point. The rat of this oscillation varies from individual to individual but, in a particular patient, the rate is fairly constant in al affected' parts of the " ody. Tremor may be caused by specifi pathological diseases (_.g. Parkinson's Disease) or be due to specific lesions in the central nervous system, or it may be of unknown origin.
- specifi pathological diseases _.g. Parkinson's Disease
- tremor can be seen as a modification of the basic electrophysiological properties of cells comprising the central nervous system.
- cer ⁇ tain compounds such as aliphatic alcohols
- these compounds when used in small amounts, do not affect the high-threshold calcium conductance.
- the inventors have found that although lower alkyl alcohols have a blocking effect, the higher alcohols do so at extraordinarily low concentrations.
- Requena et al state that octanol inter- feres with the ability of the calcium ions to leave the cell by crossing the cell membrane. This phenomenon is unrelated to blockage or non-blockage of calcium channels (high- or low threshold) .
- a calcium channel is a passive transport mechanism by which calcium ions move down their elec ⁇ trochemical gradient.
- calcium concentration is low inside the- cell (e.g., 10 ⁇ 7 M) and high in the extracellular medium (e.g., 10 ⁇ 3 M) and so a calcium channel allows calcium to go into the cell.
- a calcium pump an entirely different mechanism which trans- 0 ports calcium against a concentrational gradient (from the low concentration inside to the high concentration outside) .
- An ion pump is therefore an active membrane structure, usually an enzyme (e.g., sodium ATPase) which requires energy (ATP: adeno- sine triphosphate) to carry ions across the membrane.
- an enzyme e.g., sodium ATPase
- ATP adeno- sine triphosphate
- Beta-adrenergic blockers such as propranolol hydrochloride and its derivatives. These drugs act via a mechanism totall different from that of the present invention, and affect muscl cells as opposed to neuron cells. Beta-adrenergic blocker cause a myriad of side-effects (e.g. , bronchodilation, light
- 35 preferred alcohol concentrations primarily only LTCC would b affected and a larger patient population could be treated usin the present invention instead of beta-adrenergic blockers.
- use of alcohols in accordance with the present invention could be made in conjunction with use of beta-block- ers.
- the present invention has several objects including, but not limited to, the following:
- tremor to provide a method and composition for treating different types of tremor including, but not limited to, en ⁇ hanced physiological tremor, essential tremor, severe essential tremor, and rubral tremor.
- tremor including, but not limited to, en ⁇ hanced physiological tremor, essential tremor, severe essential tremor, and rubral tremor.
- Parkinson's tremor which can be used as an adjunct to conven ⁇ tional therapy.
- Fig. 1 is a diagram serving as an illustration of the action potential obtained by electrically stimulating I.O. cells.
- Fig. 2 is a perspective view in two sections of a brain slice perfusion and recording system in accordance with the present invention.
- Fig. 3 is a number of oscilloscope tracings and a graph showing the effect of alcohol on the low-threshold calcium con ⁇ ductance and action potential.
- Fig. 4 is an oscilloscope tracing obtained by stimulat ⁇ ing I.O. cells with incremental increases in current both i the presence and absence of 10 ⁇ 4 M butanol.
- Fig. 5 is a series of oscilloscope tracings, A throug H, showing the I.O. cell response to an external square curren injection, in the presence of 0.1% pentanol; in the presence o 0.05% pentanol; in the presence of 0.25% butanol; in the pre sence of 0.5% propanol.
- Fig. 6 (A and B) is a series of oscilloscope tracing of the ionic current obtained from I.O. cells at differen voltage amplitudes applied using a voltage clamp technique.
- the graph on the right (C) is a plot of the difference in thi ionic current (between the alcohol and the control state) against the applied external voltage.
- Fig. 7 is a series of oscilloscope tracings of th voltage response across the cell membrane in the absence (A)
- Fig. 8 is an oscilloscope tracing of electromyogram (left) and autocorrelograms (right) obtained from control o harmaline-treated rats.
- the present invention is directed to a method fo blocking or reducing the low-threshold calcium conductance i mammalian cell membranes comprising exposing said cells to a LTCC-blocking agent (such as an aliphatic alcohol) at a concen tration sufficient to block or reduce said low-threshold cal cium conductance selectively.
- a LTCC-blocking agent such as an aliphatic alcohol
- the agent will b used at a concentration sufficiently low so that it does no affect the high-threshold calcium conductance of said cells.
- Preferred agents are aliphatic alcohols and particularly pre ferred are C 3 -C 10 alkyl alcohols.
- Another aspect of the present invention is directed t a method for inhibiting the manifestation of tremor in th muscle cells of a mammal comprising exposing central nervou system cells of said mammal to an LTCC-blocking agent (such a an aliphatic alcohol) at a concentration sufficient to imped transmission of a tremor signal from said central nervous sys tem cells to said muscle cells.
- an LTCC-blocking agent such a an aliphatic alcohol
- the amount of sai agent will be insufficient to interfere with the high-threshol calcium conductance in either type of cell.
- compositions useful in blocking LTCC or in inhibiting tremor comprising an effective amount of an LTCC-blocking or a tremor- inhibiting agent and a physiologically acceptable carrier or diluent.
- the present inventors have discovered that certain agents, such as aliphatic alcohols, block or reduce specifi ⁇ cally the so-called low-threshold calcium channel in mammalian cells, and in mammalian central neurons in particular.
- This channel is known to be an important component in modulating the frequency of electrical discharges in central nuclei such as the inferior olive or thalamus.
- These agents may be used in amounts sufficient to block or reduce specifically the LTCC, without (measurably) affecting the HTCC which is involved in dendritic action potentials, in certain components of the myocardium action potential, and in synaptic transmission.
- the present invention provides a method for selectively blocking or reducing LTCC and thereby permits isolation of the HTCC.
- One use of the present invention is, therefore, in iso ⁇ lating and studying HTCC unencumbered by the LTCC, i.e. in a manner akin to that using tetrodotoxin to block sodium conduct- ance.
- the LTCC plays a basic role in the clocking properties of the brain, and pro ⁇ vides a basic frequency (continuous) for the coordination of movement (which is a series of discontinuous motions) .
- Tremor has been described as an exacerbation of the basic frequency of oscillation to the point of interference with the coordination of movement (Llinas, R.R. in Movement Disorders: Tremor (Findley, L.J. & Capildeo, R. Eds) pp. 165-182 Macmillan 1984, incorporated in this application by reference) .
- the present inventors have .further found that the same agents (e.g., aliphatic alcohols) used in the same low amounts also inhibit the symptoms of tremor.
- the importance of this finding is evident from the fac that enhanced physiological tremor, essential tremor, and th tremor produced in senile patients can be completely incapac itating.
- the method of the present invention is advantageou in that it does not affect HTCC of either the muscle or th neuron cells.
- the present invention provides a method fo treating the symptoms of tremor by acting on the central ner vous system rather than on the muscle that exhibits the tremo response, without affecting the neuron or muscle cell function that are associated with HTCC.
- the prese invention can be used with all cells that possess low-threshol calcium channels.
- aliphatic alcohol are effective in blocking LTCC and in inhibiting tremor extraordinarily low concentrations, which substantially reduc the risk of side-effects. Therefore, generally speaking, t lower the effective amount of particular alcohol, the mo desirable its use.
- the degree of inhibition of the low-conductance calci channels appears to be related to the molecular weight of t alcohol (as well as the amount used) .
- the molecular weig increases, the effective concentration of the alcohol necessa to reduce the low-conductance potential decreases.
- t calcium-conductance inhibition is greater as the alcohol m lecular weight is increased. This is demonstrated in detail Example 6, below, where it is shown that pentanol, at a 5-fo lower concentration, was more effective than propanol in i hibiting LTCC (compare Fig. 5, H with E) .
- the increasing Ca-channel-blocki ability of the higher alcohols may be due to many differe factors, such as the increasing hydrophobicity of alcohols wi increasing length of the hydrocarbon chain, and/or the lipid solubility of such alcohols.
- the operability of the present invention has been dem ⁇ onstrated on living mammalian cells both in vitro and in vivo.
- the absence of any abnormal behavior in the test ani ⁇ mals employed to demonstrate the operability of the present invention provides a further strong indication that the risk of undesirable side-effects is very small.
- the cells are exposed to a culture or perfu- sion medium containing a LTCC-blocking effective amount of the alcohol.
- This amount is preferably sufficiently small so as not to interfere with HTCC.
- ⁇ ⁇ nd - not detectable 1. (or the parenterally administered compositions) employed in the experiments of the present inventors were made up by mixing an amount of alcohol with an amount of medium sufficient to make up a 10" 6 M solution, if all of the alcohol were soluble in the
- alcohols When alcohols are administered in vivo they can generally be administered orally, or parenterally in solid or liquid form with or without a carrier or diluent. Of course, particularly in case of oral administration, allowances should be made in the amount thus administered for any amount
- the amount of the alcohol should be calculated to produce an effective LTCC-blocking (or LTCC-reducing) alcohol
- the concentration of the alcohol in the composition administered to the mammal will fall in the range at which it will produce the requisite alcohol concentration in the blood, (or in the cere-
- the alcohol will be admini ⁇ stered in a composition, also comprising a physiologically acceptable carrier or diluent. Ringer's solution or isotonic saline are preferred diluents for parenteral administration.
- octyl alcohol in an amount sufficient to generate a blood or CSF (or perfusion medium) concentration of at most 10"" 6 M.
- the amount of the LTCC-blocking or tremor-inhibiting agent will vary according to the activity of the particular agent employed and according to the mode of administration.
- the frequency of administration may also vary according to the extent of the tremor symptoms and according to how often thoy occur.
- LTCC rebound spike was generated by each of the three methods involved in the operational definition given in the background section of the present application. Regardless of the method (voltage clamp, square current injection or depo- larization voltage) used to generate the RS, alcohols block this response, indicating that a single mechanism is at work and demonstrating the operability of the present invention.
- Example l Tissue Preparation Adult Hartley guinea pigs (400-600g) from Camm Research Institute, Wayne, New Jersey were decapitated, after ethe anesthesia, using a small animal guillotine. Immediately there after, two longitudinal sections were made along the latera edge of the squamous portion of the occipital bone. The resul ting bone slab was cut transversely, and pulled caudalward t expose the cerebellum and brain stem.
- the brain stem was swiftly removed and placed i aerated Ringer's solution (containing 124 mM NaCl; 5 mM KC1 1.2 mM KH 2 P0 ; 2.4 mM CaCl 2 ; 1.3 mM MgS0 4 ; 2.6 mM NaHC0 3 and 1 mM glucose) at about 5°C.
- i aerated Ringer's solution containing 124 mM NaCl; 5 mM KC1 1.2 mM KH 2 P0 ; 2.4 mM CaCl 2 ; 1.3 mM MgS0 4 ; 2.6 mM NaHC0 3 and 1 mM glucose
- the brain slice was placed on a Sylgard plate 11 Corning Glass, Corning, N.Y.) at the bottom of the recording chamber 2 and secured with a bipolar stimulating electrode 5 pressing lightly on the brain tissue.
- the chamber was main ⁇ tained at 37 ⁇ C by a surrounding water bath 6, which kept it at the same temperature as the perfusing solution (37 ⁇ C) .
- the saline solution itself was temperature-regulated by passage through a heat exchanger 7 at 37 ⁇ C (Fig. 2A) .
- the standard perfusion fluid was Ringer's solution; this medium was used during cutting and incubating, and during most of the recording time. Ringer's solution provided excel- lent pH-buffering properties at different temperatures. This was especially significant during the sectioning and incubation periods.
- alcohols were added to the perfusion fluid by direct application into chamber 1 via the perfusion solution.
- Direct stimulation of the inferior oliv cells was implemented with a high-input impedance (10-- ohms) bridge amplifier. Capacity compensation allowed a frequenc response of 10-15 kHz, depending on the microelectrode proper ties.
- Example 4 Blockage of Low-Threshold Calcium Conductance Wit Octanol
- Figure 3A-C (upper traces) shows intracellula recordings from inferior olivary (I.O.) nuclear cell demonstrating the typical calcium-dependent action potentia after the sodium-dependent spike was blocked with 10 ⁇ 6 tetrodotoxin (Sigma Chemical Co., St. Louis, MO).
- I.O. cells were isolated as in Example l an recordings were made as in Example 3.
- the upper row in A, B, (controls conducted in the absence of octanol) shows the LTC rebound spike as the high spike.
- the HTCC is not presen because the applied current is subthreshold for the HTCC.
- Th records in the lower row in A, B, C show a complete blockage o the calcium-dependent rebound potentials due to the blockage o LTCC by the application of 10 ⁇ 6 M octanol to the bath.
- This i also shown in the "action potential” v. "membrane potential graph D on the right of Figure 3 which demonstrates the rate o rise of the low-threshold action potential versus membrane pol arization in the control records (closed circles) .
- the lowe set of data points (squares) indicates the rate of rise of th low-threshold action potential after octanol at 10 ⁇ 6 M is adde to the perfusion medium.
- the difference between the contro and the octanol graph is a measure of the blocking effect o 10 ⁇ 6 M octanol on LTCC.
- Figure 4A depicts the action potential observed in I.O neurons under conditions where the sodium channel has bee blocked by the addition of tetrodotoxin. Incremental increase in injected square current are applied producing action poten tial responses shown in Fig. 4A. When a sufficient threshol value is reached, the characteristic spikes of the action po tential are produced. The first (left-most) spike is the high threshold calcium spike and the second (or rebound) spike i the low-threshold calcium spike.
- Figure 4B depicts the resultant action potential upon the administration of 10 M butanol and the inhibition of the LTCC or rebound spike. This effect occurs in the absence of any effect on the HTCC.
- Example 6 Comparison of the LTCC-Blocking Ability and Relative Efficacy of Alcohols
- Figure 5 depicts a series of tracings, A through H, showing the I.O. cell response to an external square current injection following polarization of the membrane, both in the absence (graphs A-D) and in the presence (graphs E-H) of dif ⁇ ferent types and different concentrations of alcohol.
- Th reduction in the ratn of rise of the spike first differential of the voltage, shown in the middle record
- This Figure illus trates that higher alkyl alcohols are more effective than lowe alcohols or, conversely, that higher alkyl alcohols are a effective but at lower concentrations.
- the upper trace de picts the action potential observed across the membrane.
- Th middle trace shows the rate of change in the action potentia through the cell membrane.
- the bottom trace shows th magnitude of the injected current.
- Example 7 Voltage Clamp Study of Octanol-Induced Blockage of LTCC
- the results presented in this Example were recorded i I.O. neurons from different parts of the nucleus. These con ductances were characterized by an inward current having a ver low threshold (-70mV) which could be observed after the sodiu conductance had been blocked by the addition of tetrodoxin an the potassium conductance had been blocked by addition of te traethyl-ammonium and cesium.
- the ioni current across the membrane was measured after a voltage clam pulse was applied across the membrane.
- a current In order to maintai the voltage constant, a current must be injected into the cell This current is equal and opposite to the ionic current acros the membrane and thus can be used to determine the size an duration of the ionic current directly.
- the ionic curren generates under normal conditions (not voltage-clamped) th low-threshold calcium spike.
- FIGS. 6A and 6B show the amplitude an time course of the low-threshold calcium ionic currents fol lowing depolarization steps of 5mV from a holding potential o -80mV to a value of -45mV.
- These records represent the differ ence in ionic current observed in the control state minus tha from the passive ionic current observed after octanol was in troduced in the bath.
- the records A and B illustrat those ionic currents that were blocked by octanol at 10 " - (wjr.'.-h completely blocked the ionic conductance) .
- An inwar currant is first observed at -65mV.
- FIG. 6C depicts a plot of ionic current against the applied external voltage. The value of this current is the difference between the ionic currents before and after octanol treatment. This difference is maximum at a membrane voltage between about -45 and about -60 millivolts. This graph charac- terizes the voltage dependence of the LTCC.
- the graph D on the right of Figure 7 shows the depen dence of the rebound calcium spike on the amplitude of th injected hyperpolarizing current.
- octanol was administered to rats. Alcohols were diluted with physiological saline and sonicated to ensur mixing to create a concentration of alcohol around the neuron equivalent to 10 —6M (at most) . The alcohol was administere intraperitoneally at a concentration of 1 to lOmM. Tremo state was induced in the rats by injection of har aline, a alkaloid derived from Pegamus harmala which has been known t induce tremor in mammals. (Harmaline is available from Sigm
- the thus-induced tremor has periodicity similar to physiological or enhanced physiological tremor.
- I.O. cells have been found to be responsible for trans mitting the tremor signal to mammalian muscles, since destruc tion of I.O. cells abolishes tremor in harmaline-treated ani als (Llinas, et al. , Science 190: 1230-1231, 1975). There fore, study of I.O. cells of mammals in which tremor has bee induced with harmaline provides an excellent in vivo model fo studying tremor response in mammals.
- Tremor can be measured by electromyogram conducted ac cording to Buchthal, F. , et al., Acta Phvsio. Scand. 39: 83 104, 1957, incorporated by reference.
- the electric activity in the left platysma muscle o 278 to 300 g Sprague-Dawley rats was recorded differentially using two teflon coated stranded silver wires (2 mm of wire exposed) placed i the belly of the muscle.
- the electromyogram (emg) was recorde throughout the experiment and the data were analyzed at th following times: (1) before administration of any drugs; (2) after intraperitoneal injection of 15 mg/kg harmaline; and (3) after subsequent intraperitoneal injection of 2 ⁇ c of 10 mM 1 octanol (99% octanol sonicated in physiological saline) give to create a concentration of 9.3 micrograms/g of rat bod weight or 7 x 10 -5M m. the vi.ci.ni.ty of the neuronal cells (i all the alcohol was to be absorbed, and none metabolized) .
- Th emg was smoothed and differentiated (to further decrease th background noise) using a Nicolet Explorer Model 4060 digita oscilloscope (from Nicolet Instrument Corp. , Madison, WI) . This instrument was also used to obtain autocorrelograms o 800-ms sections of the emg's. Autocorrelograms were obtaine by superimposing 800-ms sections of the emg signals. Th emerging patterns (if any) can be used to determine frequenc and amplitude characteristics of the emg that may not b readily discernible from the naked emg signal.
- Control emg's demonstrated occasional periods of lo amplitude oscillations (upper left, Figure 8) .
- Autocorrelatio revealed an 8 Hz periodicity in t e emg, but with a low cor relation function (upper right, Figure 8) .
- the animals demonstrated periods of generalized tremor. The emg recorded during such a period is illustrated in the middle left-hand trace of Figure 8; the tremor had a dominant frequency of 7.5 Hz, close to the control value, and showed a high degree of autocorrelation (middle right-hand trace).
- the tremor stopped and the baseline muscle activity was reduced below control levels (bottom left-hand trace) .
- the dominant frequency was at 10 Hz, and the emg amplitude was so low during this period that it was necessary to amplify the signal in order to obtain an auto ⁇ orrelogram (bottom, right) .
- a 10-Hz periodicity is evident (as shown by the arrows)
- the 60-Hz frequency due to line voltage (amplifier noise) is the dominant correlation due to the large gain. If the bottom right signal had been ampli- fied only to the extent of the two upper autocorrelograms, it would have appeared as a straight line.
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Abstract
Procédé permettant de bloquer partiellement ou totalement les canaux de calcium de seuil dans les membranes cellulaires. Le procédé consiste à exposer les cellules à une quantité efficace d'un agent bloquant la conductance de calcium à seuil faible.Method for partially or totally blocking threshold calcium channels in cell membranes. The method involves exposing the cells to an effective amount of a low threshold calcium conductance blocking agent.
Description
Claims
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
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US83708886A | 1986-03-06 | 1986-03-06 | |
US837088 | 1986-03-06 |
Publications (2)
Publication Number | Publication Date |
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EP0260312A1 true EP0260312A1 (en) | 1988-03-23 |
EP0260312A4 EP0260312A4 (en) | 1989-11-07 |
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EP19870902204 Withdrawn EP0260312A4 (en) | 1986-03-06 | 1987-02-23 | Method for blocking calcium channels. |
Country Status (5)
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EP (1) | EP0260312A4 (en) |
JP (1) | JPS63500525A (en) |
AU (1) | AU7206087A (en) |
IL (1) | IL81720A0 (en) |
WO (1) | WO1987005214A1 (en) |
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US2903396A (en) * | 1957-05-08 | 1959-09-08 | Ciba Pharm Prod Inc | Therapeutic compositions and method for treating parkinsonism |
US3041237A (en) * | 1960-08-15 | 1962-06-26 | Warren Teed Products Company | Systemic composition for the reduction of inflammation and edema and method of administering |
US3778509A (en) * | 1971-05-28 | 1973-12-11 | Cotton Inc | Control of fungi on cotton plants |
-
1987
- 1987-02-23 WO PCT/US1987/000413 patent/WO1987005214A1/en not_active Application Discontinuation
- 1987-02-23 JP JP62502074A patent/JPS63500525A/en active Pending
- 1987-02-23 AU AU72060/87A patent/AU7206087A/en not_active Abandoned
- 1987-02-23 EP EP19870902204 patent/EP0260312A4/en not_active Withdrawn
- 1987-03-01 IL IL81720A patent/IL81720A0/en unknown
Non-Patent Citations (9)
Title |
---|
Biochem. Pharmacology, Vol. 32, No. 6, 1983, pages 963-969, Pergamon Press Ltd., GB; M.L. MICHAELIS et al.: "Alcohol and local anesthetic effects on Na dependent Ca2+ fluxes in brain synaptic membrane vesicles", Abstract; figures 2,3,4a,4b; page 965, left-hand column, line 1 - page 965, right-hand column, line 47; page 967, right-hand column, line 50 - page 968, left-hand column, line 2. * |
Can. J. Physiol. Pharmacol., Vol. 60, No. 5, May 1982, pages 610-614, Nat. Research Council of Canada, J.G. SINCLAIR et al.: "Ethanol effects on the olivocerebellar system", Summary, page 610, section "Introduction"; figure 2; page 612, right-hand column, lines 12-16; page 613, right-hand column, line 10 - page 614, left-hand column, line 6. * |
Fed. Proc., Vol. 36, No. 3, 1977, page 353, Abstract 444; W.D. KNOWLES et al.: "Effects of amesthetics on cerebellar neuroms recorded chromically after harmaline", whole Abstract. * |
J. Pharm. Exp. Ther., Vol. 225, No. 8, 1983, pages 571-575, US; S.W. LESLIE et al.: "Inhibition of fast- and slow-phase depolarization-dependent synaptosomal calcium uptake by ethanol", Abstract; table 1; pages 572-573, section "Results"; page 573, right-hand column, line 18 - page 574, left-hand column, line 3. * |
Life Sciences, Vol. 34, January 1984, pages 49-56, Pergamon Press, US; M.S. RAPPAPORT et al.: "Ethanol effects on harmaline-induced tremor and increase of cerebellar cyclic GMP", Summary; figures 1,2; pages 51,52, section "Results", page 54, last paragraph. * |
Pfluegers Arch, Vol. 414, No. 1, May 1989, pages 31-36, European Journal of Physiology, Springer Verlag, DE, C.M. SINTON et al.: "The effectiveness of different isomers of octanol as blockers of harmalinc-induced tremor", Abstract; page 34, left-hand column, line 9 - page 34, right-hand column, line 27; page 36, left-hand column, lines 7-47. * |
See also references of WO8705214A1 * |
Soc. Neurosci. Abstr., Vol. 12, No. 1, November 1986, page 174; Abstr. 49.3, R. LLINAS et al.: "Specific blockage of the low threshold calcium channel by high molecular weight alcohols", whole Abstract. * |
Therapie, Vol. 32, 1977, pages 355-364; O. SPREUX-VAROQUAUX et al.: "Ethanol et tremblements experimentaux chez la souris et le rat", Summary; figures 2,3; page 362, lines 24-33. * |
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Publication number | Publication date |
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JPS63500525A (en) | 1988-02-25 |
EP0260312A4 (en) | 1989-11-07 |
WO1987005214A1 (en) | 1987-09-11 |
AU7206087A (en) | 1987-09-28 |
IL81720A0 (en) | 1987-10-20 |
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