CA2195779A1 - Composition and method for protection against drug dependency - Google Patents
Composition and method for protection against drug dependencyInfo
- Publication number
- CA2195779A1 CA2195779A1 CA002195779A CA2195779A CA2195779A1 CA 2195779 A1 CA2195779 A1 CA 2195779A1 CA 002195779 A CA002195779 A CA 002195779A CA 2195779 A CA2195779 A CA 2195779A CA 2195779 A1 CA2195779 A1 CA 2195779A1
- Authority
- CA
- Canada
- Prior art keywords
- agonist
- hexane
- acts
- substance
- acid
- 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.)
- Abandoned
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D235/00—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, condensed with other rings
- C07D235/02—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, condensed with other rings condensed with carbocyclic rings or ring systems
-
- 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
-
- 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
- A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
- A61K31/19—Carboxylic acids, e.g. valproic acid
- A61K31/195—Carboxylic acids, e.g. valproic acid having an amino group
- A61K31/197—Carboxylic acids, e.g. valproic acid having an amino group the amino and the carboxyl groups being attached to the same acyclic carbon chain, e.g. gamma-aminobutyric acid [GABA], beta-alanine, epsilon-aminocaproic acid, pantothenic acid
- A61K31/198—Alpha-aminoacids, e.g. alanine, edetic acids [EDTA]
-
- 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
- A61K31/21—Esters, e.g. nitroglycerine, selenocyanates
- A61K31/215—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids
-
- 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
- A61P25/30—Drugs for disorders of the nervous system for treating abuse or dependence
-
- 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
- A61P25/30—Drugs for disorders of the nervous system for treating abuse or dependence
- A61P25/34—Tobacco-abuse
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C229/00—Compounds containing amino and carboxyl groups bound to the same carbon skeleton
- C07C229/46—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino or carboxyl groups bound to carbon atoms of rings other than six-membered aromatic rings of the same carbon skeleton
- C07C229/50—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino or carboxyl groups bound to carbon atoms of rings other than six-membered aromatic rings of the same carbon skeleton with amino groups and carboxyl groups bound to carbon atoms being part of the same condensed ring system
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C62/00—Compounds having carboxyl groups bound to carbon atoms of rings other than six—membered aromatic rings and containing any of the groups OH, O—metal, —CHO, keto, ether, groups, groups, or groups
- C07C62/30—Unsaturated compounds
- C07C62/38—Unsaturated compounds containing keto groups
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C2602/00—Systems containing two condensed rings
- C07C2602/02—Systems containing two condensed rings the rings having only two atoms in common
- C07C2602/14—All rings being cycloaliphatic
- C07C2602/18—All rings being cycloaliphatic the ring system containing six carbon atoms
Abstract
The present invention provides the use of an agonist which acts at negatively coupled cAMP-linked metabotropic glutamate receptors for the manufacture of a medicament for the treatment of substance dependence disorders.
Description
~ W096/04900 2 1 9 5 7 7 9 PCT~S95/10317 C~",~ c~ n and method for ~,~ut~ iùl~ a~ainst dru~ da,~e"de"~.
The present invention relates to treatments for substance dependence.
Substance ~rprn~rnre is a major problem, both for individuals suffering from it, and for society at large. At the individual level, the condition is characterized by a need for repeated, and often increasing doses of a substance.
At the societal level, the condition is associated, with some substances, with increased levels of crime, inr1n~inr theft and crimes of violence, as sufferers seek to obtain supplies of the substance.
Individuals dependent on a snh~t~nrP find that they have to continue taking the substance, even though it produces harmful effects in them. They may become tolerant to the substance, which means that they need to take greatly increased amounts, for example ten times the amount they originally took, in order to achieve the same effect.
Withdrawal of the substance brings about a variety of undesirable behavioral and physiological changes, inrl~ing craving for the substance, anxiety and irritability.
In the ~ n central nervous system (CNS), the transmission of nerve impulses is controlled by the interaction between a neurotransmitter, that is released by a sending neuron, and a surface receptor on a receiving neuron, causing excitation of this receiving neuron.
L-Glutamate, which is the most ~hnn~nt neurotranSmitter in the CNS, mediates the major excitatory pathway in mammals, and is referred to as an excitatory amino acid (EAA). The receptors that respond to glntr~-to are called excitatory amino acid receptors ~ (EAA receptors). See Watkins & ~vans, Ann. Rev.
Pharmacol. ~oxicol., 21, 165 (1981); Mrn~rh~n, Bridges, and Cotman, Ann. ~ev. Pharmacol. Toxicol., 29, 365 (1989); Watkins, Krogsgaard-Larsen, and Honore, Trans.
Pharm. Sci., 11, 25 (1990). The excitatory amino acids PC~IUS 9 5 ~ 1~ 31 7 ~ X-9329D 2 ~ ~ 5 7 7 9 IPEA]US 1 ~ AUG 1996 are of great physi~logi~Al importance, playing a role in a variety of physiological processes, such as long-term potentiation (l~Arn;ng and memory), the development of synaptic plasticity, motor control, respiration, cardiovascular regulation, emotional states and sensory perception.
Excitatory amino acid receptors are classified into two general types. Receptors that are directly coupled to the opening of cation rhAnn~l R in the cell membrane of the neurons are termed "ionotropic." This type of receptor has been subdivided into at least three subtypes, which are defined by the depolarizing actions of the selective agonists N-methyl-D-aspartate (NMDA), ~-amino-3-hydroxy-5-methylicnYA~ol~-4-propionic acid (AMPA), and kainic acid (KA). The second general type of receptor is the G-protein or second messenger-linked "metabotropic" excitatory amino acid receptor. This second type is coupled to multiple second messenger systems that lead to ~nhAn~e~ pho9rhn; nnc; ti~e hydrolysis, activation of phosphnlirAce D, increases or decreases in cAMP formation, and changes in channel function. Schoepp and Conn, Trends in Pharmacol. Sci., 14, 13 (1993). Both types of receptors appear not only to mediate normal synaptic transmission along excitatory pathways, but also participate in the ~;fi~Ation of synaptic connections during development and throughout life. Schoepp, Bockaert, and S1A~CZ~k, Trends in Phar~acol. Sci., 11, 508 (1990); McDonald and Johnson, Brain Research Reviews, 15, 41 (1990).
The metabotropic glutamate receptors are a highly heteL.g~ ~ q family of glutamate receptors that are linked to multiple second-messenger pathways. Generally, these L~c~L~L~ function to modulate the presynaptic release of glutamate, and the postsynaptic sensitivity of the neuronal cell to glutamate excitation. The metabotropic glutamate receptors (mGluR) have been phArr--olngically divided into two subtypes. One group of receptors is positively coupled _ _ _ _ _ _ , .. .. . . . .. . _ . _ ~ W096/04900 ~l 9~779 PCT~S95110317 to phospholipase C, which causes hydrolysis of cellular phosphoinositides (PI). This first group are termed PI-linked metabotropic glutamate receptors. The second group of receptors is negatively coupled to adenyl cyclase, which prevents the forskolin-stimulated a,- 11 At~ i nn of cyclic adenosine monophosphate (cAMP). Schoepp and Conn, Trends Pharmacol. Sci., 14, 13 (1993). Receptors within this second group are termed cA~P-linked metabotropic glutamate receptors.
There are many different substances on which individuals may become ~pPn~nt. These include opiates, h~n7n~iA7epines, nicotine, cocaine and ethanol.
Nicotine dependence. which is induced through smoking, affects hundreds of millions of people around the world. For many, it leads to illness and premature death. Stopping smoking (smoking cessation) may evoke a range of symptoms in ~pPn~nt individuals, inrlu~ing craving, depression, anxiety, difficulty in concentrating and weight gain.
A variety of treatments are available for smoking ~c~Atin~, ;nrlll~ing counseling, hypnosis, aversion ~nn~ i t; ~n i ng, relaxation training, acupuncture, and nicotine replacement therapy. However, in spite of the avAilAhility of these tr~A ~, and the widespread knowledge of the harmful side effect of smoking, many smokers fail to give up smoking. There is therefore a need for new treatments for smoking cessation.
senzO~iA7~pinP dependence, such as diazepam dependence, arises through the use of the benzodiazepines as pharmaceuticals to treat other disorders. The dependence-in~ ing properties of the bPn7o~iA7epines limits their therapeutic use. Withdrawal produces symptoms such as anxiety, irritability, inq~ iA and impaired uu"c~"~L~tion.
There is therefore a need for new treatments for the treatment of benzodiazepine withdrawal.
Animal models for the treatment of nicotine and diazepam withdrawal have been described in Helton et ~1_;
Psychopharmacology (1993), 113:205-210 and RA -A~n ~ ~1_;
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ . _ .
The present invention relates to treatments for substance dependence.
Substance ~rprn~rnre is a major problem, both for individuals suffering from it, and for society at large. At the individual level, the condition is characterized by a need for repeated, and often increasing doses of a substance.
At the societal level, the condition is associated, with some substances, with increased levels of crime, inr1n~inr theft and crimes of violence, as sufferers seek to obtain supplies of the substance.
Individuals dependent on a snh~t~nrP find that they have to continue taking the substance, even though it produces harmful effects in them. They may become tolerant to the substance, which means that they need to take greatly increased amounts, for example ten times the amount they originally took, in order to achieve the same effect.
Withdrawal of the substance brings about a variety of undesirable behavioral and physiological changes, inrl~ing craving for the substance, anxiety and irritability.
In the ~ n central nervous system (CNS), the transmission of nerve impulses is controlled by the interaction between a neurotransmitter, that is released by a sending neuron, and a surface receptor on a receiving neuron, causing excitation of this receiving neuron.
L-Glutamate, which is the most ~hnn~nt neurotranSmitter in the CNS, mediates the major excitatory pathway in mammals, and is referred to as an excitatory amino acid (EAA). The receptors that respond to glntr~-to are called excitatory amino acid receptors ~ (EAA receptors). See Watkins & ~vans, Ann. Rev.
Pharmacol. ~oxicol., 21, 165 (1981); Mrn~rh~n, Bridges, and Cotman, Ann. ~ev. Pharmacol. Toxicol., 29, 365 (1989); Watkins, Krogsgaard-Larsen, and Honore, Trans.
Pharm. Sci., 11, 25 (1990). The excitatory amino acids PC~IUS 9 5 ~ 1~ 31 7 ~ X-9329D 2 ~ ~ 5 7 7 9 IPEA]US 1 ~ AUG 1996 are of great physi~logi~Al importance, playing a role in a variety of physiological processes, such as long-term potentiation (l~Arn;ng and memory), the development of synaptic plasticity, motor control, respiration, cardiovascular regulation, emotional states and sensory perception.
Excitatory amino acid receptors are classified into two general types. Receptors that are directly coupled to the opening of cation rhAnn~l R in the cell membrane of the neurons are termed "ionotropic." This type of receptor has been subdivided into at least three subtypes, which are defined by the depolarizing actions of the selective agonists N-methyl-D-aspartate (NMDA), ~-amino-3-hydroxy-5-methylicnYA~ol~-4-propionic acid (AMPA), and kainic acid (KA). The second general type of receptor is the G-protein or second messenger-linked "metabotropic" excitatory amino acid receptor. This second type is coupled to multiple second messenger systems that lead to ~nhAn~e~ pho9rhn; nnc; ti~e hydrolysis, activation of phosphnlirAce D, increases or decreases in cAMP formation, and changes in channel function. Schoepp and Conn, Trends in Pharmacol. Sci., 14, 13 (1993). Both types of receptors appear not only to mediate normal synaptic transmission along excitatory pathways, but also participate in the ~;fi~Ation of synaptic connections during development and throughout life. Schoepp, Bockaert, and S1A~CZ~k, Trends in Phar~acol. Sci., 11, 508 (1990); McDonald and Johnson, Brain Research Reviews, 15, 41 (1990).
The metabotropic glutamate receptors are a highly heteL.g~ ~ q family of glutamate receptors that are linked to multiple second-messenger pathways. Generally, these L~c~L~L~ function to modulate the presynaptic release of glutamate, and the postsynaptic sensitivity of the neuronal cell to glutamate excitation. The metabotropic glutamate receptors (mGluR) have been phArr--olngically divided into two subtypes. One group of receptors is positively coupled _ _ _ _ _ _ , .. .. . . . .. . _ . _ ~ W096/04900 ~l 9~779 PCT~S95110317 to phospholipase C, which causes hydrolysis of cellular phosphoinositides (PI). This first group are termed PI-linked metabotropic glutamate receptors. The second group of receptors is negatively coupled to adenyl cyclase, which prevents the forskolin-stimulated a,- 11 At~ i nn of cyclic adenosine monophosphate (cAMP). Schoepp and Conn, Trends Pharmacol. Sci., 14, 13 (1993). Receptors within this second group are termed cA~P-linked metabotropic glutamate receptors.
There are many different substances on which individuals may become ~pPn~nt. These include opiates, h~n7n~iA7epines, nicotine, cocaine and ethanol.
Nicotine dependence. which is induced through smoking, affects hundreds of millions of people around the world. For many, it leads to illness and premature death. Stopping smoking (smoking cessation) may evoke a range of symptoms in ~pPn~nt individuals, inrlu~ing craving, depression, anxiety, difficulty in concentrating and weight gain.
A variety of treatments are available for smoking ~c~Atin~, ;nrlll~ing counseling, hypnosis, aversion ~nn~ i t; ~n i ng, relaxation training, acupuncture, and nicotine replacement therapy. However, in spite of the avAilAhility of these tr~A ~, and the widespread knowledge of the harmful side effect of smoking, many smokers fail to give up smoking. There is therefore a need for new treatments for smoking cessation.
senzO~iA7~pinP dependence, such as diazepam dependence, arises through the use of the benzodiazepines as pharmaceuticals to treat other disorders. The dependence-in~ ing properties of the bPn7o~iA7epines limits their therapeutic use. Withdrawal produces symptoms such as anxiety, irritability, inq~ iA and impaired uu"c~"~L~tion.
There is therefore a need for new treatments for the treatment of benzodiazepine withdrawal.
Animal models for the treatment of nicotine and diazepam withdrawal have been described in Helton et ~1_;
Psychopharmacology (1993), 113:205-210 and RA -A~n ~ ~1_;
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ . _ .
N~UL~'L~OLL 5, 154-156 (lg93). ~hese models can be used to measure the ability of a test compound to inhibit the increased startle response in an animal ~rat) following withdrawal of nicotine or diazepam.
It has now been found that a , _ fl which is an agonist that acts selectively at negativelY coupled cAME-linked rihntropic glutamate receptors is capable of reducing startle response in rats following the cessation of chronic nicotire or fliA7ep-~m exposure. It is believed that this finding portends that any agonist that acts at negatively coupled cAMP-linked metabotropic glutamate receptors will be useful for treating substance withdrawal, and indeed may be useful for providing protection against substance dependence.
According-to one aspect, therefore, the present invention provides the use of an agonist which acts at negatively coupled cANP-linked metabotropic glutamate receptors for protecting a warm blooded ma~mal from dependence on a-substance.
It has ~een found that a novel compound which is an agonist that acts selectively at negatively coupled cAMP-linked metabotropic glutamate receptors, ~+)-2-aminobicyclo[3.1.0]hexane-2,6-dicarboxylic acid is effective in the rat startle models for nicotine and diazepam withdrawal. Accordingly it is believed that any compound which acts as an agonist at negatively coupled cAMP-linked metabotropic glutamate raceptors, especially any agonist which acts selectively, will be useful for the treatment of the withdrawal or CPqsati nn of these and other dependence-producing substances. Furthermore, since withdrawal isinrir~tPly related to flPpPnflPn~e and to tolerance, it is believed that an agonist which acts at negatively coupled cAME~-linked metabotropic glutamate receptors will also be useful for the treatment of substance dependence and tolerance, and indeed generally to protect a warm blooded mammal from flPpPnflPn~e on a flPpPnflPnnP-producing substance.
~ W096/04900 2 1 9 5 7 7 q PCT~S9~10317 The ~PpPn~Pnre-producing substance may be, for example, an opiate, benzodiazepine, nicotine, cocaine or ethanol.
According to another aspect, the present invention provides the use of an agonist which acts at negatively coupled cAMP-linked metabotropic glutamate receptors for the treatment of drug tolerance, withdrawal or cessation.
According to yet another aspect, the present invention provides the use of an agonist which acts at negatively coupled cA~P-linked metabotropic glutamate receptors for the treatment of smoking cPq~ti nn .
The particular dose of agonist administered will of course be ~PtPrminpd by the particular circumstances surrounding the case, ;nrln~inr the activity of the particular agonist administered, the route of administration, the particular condition being treated, and similar considerations. The agonist can be administered by a variety of routes inr1ll~ing oral, rectal, trAnc~Prr-l, subcutaneous, intravenous, intramuscular, or intranasal routes. Alternatively, the agonist may be administered by continuous infusion. A
typical daily dose will contain from about O.OOl mg/kg to about lO0 mg/kg of the agonist. Preferably, daily doses will be about 0.05 mg/kg to about 50 mg/kg, more preferably from about O.l mg/kg to about 20 mg/kg.
According to preferrea aspect, the present invention provides the use of an agonist as defined hereinabove for use in a method of protecting a warm-blooded mammal from ~PpPn~Pnre on a dependence-producing pharmaceutical, for example, a bPn7o~iA7~r;np such as ~;~7Pp~m. In this method, the agonist may be administered before said phArr--Putical is first administered, after said phArr--Plltir~l has been administered or after said phArr-rPnt;rAl has been withdrawn, or it may be co-administered with said pharmaceutical.
Agonists which act at negatively coupled cANP-linked metabotropic glutamate receptors may be PCTIUS 9 5 / 1~ 3 ~-g329D ~ 9~7~q IPEAIUS lb AUG 1996 identified using the following ~Yp~ri L. Firstly, the affinity of a test compound for metabotropic glutamate receptors may be demonstrated by the selective disrlA~ L of (15,3R)-1-aminocyclopentane-1,3-~io~rh~ylic acid-sensitive [3H]glutamate binding to rat brain cell membranes. The binding of [3H]glutamate ([3H]Glu) is conducted with crude membranes of rat forebrain as described by Schoepp and True. Schoepp and True, Neuroscience Lett., 145, 100-104 (1992); Wright, McDonald, and Schoepp, J. Neurochem., 63, 938-945 (1994). The affinity of a test ~ _od for the receptor may be expressed as the concentration of the test compound that inhibits 50i binding (ICso), or the percent dis~l~, L of [3H]Glu at a 10 yM or 100 yM
concentration of the formula I c ~. In this test, the ICso for (+)-2-aminobicyclo[3.1.0]hexane-2,6-~;o~rh~xylic acid was found to be 0.18 yM.
The ability of a test . _nd to act as an agonist at negatively coupled cAMP-linked metabotropic receptors may be measured using the following method. Test c, ullds are tested for their ability to decrease forskolin-stimulated cAMP formation in the rat h;rporP ,us and the rat cerebral cortex, using the ~-~cedul~s described in Schoepp and Johnson. Schoepp and Johnson, Neurochem. Int., 22, 277-283 (1993). In this test, (+)-2-~min~h;~yclo[3~l.o]hexane-2~6-~ie~rh~ylic acid was found to give the result shown in Table II below.
Table II. Inhibition of Forskolin-6timulated cAMP
F. i~
ECso (yM) Rat cerebral cortex .055 + .017 Rat hi,p~- ~ .036 1 .015 -9329D ~ ~ 1996 21 ~5779 The ability of negatively coupled cAMP-linked metabotropic receptor agonists to protect a warm blooded ma_mal from the effects of drug withdrawal or cessation may be demonstrated using an auditory startle model. In 5 this model, animals are dosed with a drug (nicotine or ~;?70p~m), then dosing is discontinued. This cessation of drug dosing elicits an increased startle response to auditory stimuli. Test compounds are then administered to animals to determine whether they are capable of 10 attenuating the increased startle response.
Long Evans rats t200-400 g; Harlan Sprague Dawley, Columbus, IN) were individually housed in a controlled environment on a 12 hour light-dark cycle and given free access to food (Purina Rodent Chow) and water. Rats 15 were anesthot;7ed with isoflurane and Alzet osmotic pumps (Alza Corporation) were implanted su~cutaneously.
Test c- 1 was dissolved in a vehicle of purified water and neutralized with 5N NaOH to a pH of 7-8 when appl;~hlo. Diazepam (Sigma Chom;~l Company, 20 St. ~ouis, MO) was su~pon~od in a vehicle consisting of 40% PEG 300, 10% EtOH, 2% benzyl alcohol, 1% Tween 80, and 47% purified water. Nicotine (Research Pio~hom;~
Inc., Natick, MA) was dissolved in saline. Control animals received the respective vehicle.
25 Nicotine withdrawal: Pumps were filled to deliver nicotine (6mg/kg/day s.c.), diazepam (lOmg/kg/day s.c.), test compound (0,1,3,1Omg/kg s.c.) or vehicle. Twelve days following subcutaneous implantation of pumps, rats were anesthetized with isoflurane and the pumps were 30 removed. During withdrawal (following pump removal), the auditory startle response (peak amplitude, Vmax) of individual rats was recorded using San Diego Instruments startle rhl ~ (San Diego, CA). Startle sessions consisted of a 5-minute adaptation period at a 35 background noise level of 70+2dBA immediately followed by 25 presentations of auditory stimuli (120+2d~A noise, 50ms duration) presented at 8-second intervals. Peak W096l04900 21 9577~ PCT~S9S/10317 -startle amplitudes were then averaged for~all 25 presPnt~t; nnc of stimuli for each session and all data are preseAted here as overall session means. Auditory startle rPcpnn~; ng was evaluated daily on withdrawal days 1,2,3,4 or 5. saseline startle rpcpnn~ing was evaluated prior to pump removal on day 12.
Auditory startle rPcpnn~ing was significantly increased through the first three days following cPcqstinn of chronic nicotine exposure when compared to control rats receiving water. Rats given a rep~
dose of nicotine at doses of 0.03 mg/kg, i.p. or higher did not display the same heightened startle response seen for aAimals with no nicotine rPpl~n~ ~nt.
Pretreatment with ~+)-2-aminobicyclo[3.1.0]hexane-2,6-dicarboxylic acid produced a dose-dependent hlonk~P Of the withdrawal-induced increase in startle responding as well. A significant ~ttPnn~tinn of the heightened startle was apparent at 3mg/kg, p.o. dose of the compound when compared to nicotine coAErols (ED50=0-7 mg/kg i.p.).
D~7f~m Withdrawal: Auditory startle r~Qpnn~ing was qi~ni~in~ntly increased through the first four days following cessation of chronic dia_epam exposure when compared to control rats receiving vehicle Replacement doses of 3 and lO mg/kg, i.p. ~;~7ap~m did not block the increased start~le rPqpnn~;ng and in some ;nqt~nnPR
further iAcreased reactivity indicating toleraAce. Rats which received 30 mg/kg, i.p. ~;~7Pp~m rep~ daily 60 minutes before ev~ln~tinn of startle roqpnn~;ng, did not show increased reactivity following ~;~7Pp~m C~qq~ti nn on days 1 through ~ when I ed to the ~i~7Pp~m control. Pretreatment with (+)-2-innhinyc1O[3~l~o] hexane-2,6-dicarboxylic acid blocked the expected increase in startle rpcpnn~i ng which followed cessation of diazepam exposure. 3Oses of 0.1 aAd 0.3 mg/kg, p.o. of the c _ ~ significaAtly ~ W096/04900 2 1 9 5 7 7 9 PCT~S95/10317 ~ttonn~ted enhanced startle when compared to control rPcprm~;n!J ~ED50=0.1 mg/kg, p.o. ) .
(+)-2-Aminobicyclo[3.1.0]hexane-2,6-dicarboxylic acid may be prepared by reacting carbethoxYmethyl dimethy1c-~1f~nillm bromide with 2-cyclopenten-1-one in the presence of a base, such as 1,8-diazabicyclo[5.4.0]undec-7-ene to afford ethyl 2-oxobicyclo[3.l~o]hexane-6-carboxylate. This ester may then be reacted with an aqueous solution of potassium cyanide or sodium cyanide and . illm carbonate to produce an int~L -~;Ate hydantoin, (the sucherer-sergs reaction), which is then hydrolysed using sodium hydroxide, to afford a diastereomeric mixture of diethyl 2-aminobicyclo[3.1.0]hexane-2,6-~;c~rh~ylates. The desired diastereomer may be obtained by crystallization wi'th oxalic acid. This diastereomer may then be resolved by forming a crystalline salt with (+)-di-p-toluoyl-D-tartaric acid and recovering (-)-diethyl Z-aminobicyclo[3.1.0]hexane-2,6-dicarboxylate. Xydrolysis of this diester using aqueous sodium hydroxide gives (+)-2-aminobicyclo[3.1.0]hexane-2,6-dicarboxylic acid.
Alternatively, the ethyl 2-oxobicyclo[3.1.0]hexane-6-carboxylic acid may be hydrolysed using sodium hydroxide to give 2-oxobicyclo[3.1.0]hexane-6-carboxylic acid.
This acid may then be resolved by formin~ a crystalline salt with (S)-1-phenylethylamine and recovering (+)-2-oxobicyclo[3.1.0]hexane-6-carboxylic acid. This acid may then be converted into (+)-2-aminobicyclo[3.1.0]
hexane-2,6-dicarboxylic acid by reaction with an aqueous solution of potassium cyanide or sodium cyanide and ammonium carbonate to produce an intermediate hydantoin (the sucherer-sergs reaction) followed by hydrolysis of the hydantoin using sodium hydroxide. This procedure may also be modified by performing the resolution step on the hydantoin rather than on the 2-oxobicyclo[3.1.0]hexane-6-carboxylic acid. In this W096/04900 2 1 9 5 7 7 ~ PCT~S9~10317 -case, (~ phenylethylamine has been found to be a suitable resolving agent.
The agonists are preferably formulated prior to administration in , ~ in~t;nn with one or more pharm--Pllt;r~lly-acceptable carriers, ~ lPntq, or ~ripi~ntq. The pharmaceutical formulations are prepared by known procedures using well-known and readily availa~le ingredients. In making the compositions, the active ingredient will usually be mixed with a carrier, or diluted by a carrier, or enclosed within a carrier, and may be in the form of a capsule, sachet, paper, or other nnnt~inPr, When the carrier serves as a diluent, it may be a solid, semi-solid, or liriuid material which acts as a 'vehicle, ~nipi~nt, or medium for t-h-e active ingredient. The compositions can be in the form of tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols, ni ' c cnnt~;ning, for example, up to 10% by weight of active compound, so~t and hard gelatin capsules, suppositories, sterile iniectable Snl--tinnq, dermal patch, snhcllt~n~ous implant, and sterile packaged powders.
Some examples o~ snitRhl~ carriers, excipients, and diluents include lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum, acacia, calcium phnsph~te~
alginates, trAgar~nth, gelatin, calcium silicate, microcrystalline cel1lllose, polyvinylpyrrolidone, cellulose, water syrup, methyl cellulose, methyl and propyl hydroxyhPn7oat~q, talc, r~gn~qinm stearate, stearic acid, and mineral oil. The f~ lat;nnq can additionally include lubricating agents, wetting agents (surfactants), emulsifying and suspending agents, preserving agents, sweetening agents, or flavoring agents. Compositions may be f~ l~t~ 50 as to provide r~uick, sustained, or delayed release of the active ~ W096/~900 2 ~ ~5779 P~ 17 ingredient after administration to the patient by employing procedures well known in the art.
The compositions are preferably formulated in a unit dosage form, each dosage crntAininr from about 1 mg to about 500 mg, more preferably about 5 mg to about 200 mg of the active ingredient. The term ~unit dosage form'~ refers to a physically discrete unit suitable as unitary dosages for human subjects and other mammals, each unit rnntA;ninr a predetrrmin~d quantity of active material calculated to produce the desired therapeutic effect, in association with a suitable ph~rrace1ltir~
carrier, diluent, or excipient. The following formulation examples are illustrative only and are not --int~n~ to limit the scope of the invention in any way.
2~ 95779 W096/04900 PCT~S95/10317 -Formulation ~ard gelatin capsules are prepared using the ~ollowing ingredients:
Quantity (mg/capsule) 2-Aminobicyclo[3.1.0]hexane-2,6-dicarboxylic Acid 250 Starch, dried 200 ~agnesium stearate 10 Total 460 mg 20The above ingredients are mixed and filled into hard gelatin capsules in 460 mg quantities.
Formulation 2 A tablet is prepared using the ingredients below:
ouantity 30~mg/tablet) 2-~mi nnh; cyclo[3.1.0]hexane-2,6-dicarboxylic Acid 250 35 rPl l n l nse, microcrystalline400 Silicon dioxide, fumed 10 Stearic acid . 5 Total 665 mg The c ~ Pntc are blended and compressed to form tablets each weighing 665 mg.
~ W096104900 2 1 9 ~ 7 7 9 PCT~S95/10317 Formulation 3 An aerosol solution is prepared ~ntAining the following , ~ntc:
. .
Weight 2-Aminobicyclo[3.1.0]hexane-2,6-~iCArh~xylic Acid 0.25 Ethanol 29 . 75 Propellant 22 70 . 00 ~chlorodifluoromethane) Total 100.00 The active compound is mixed with ethanol and the mixture added to a portion of the PropPl 1 Anr 22, cooled to -30~C and transferred to a filling device. The re~uired amount is then fed to a stainless steel r~ntAinPr and diluted with the 1 ;n~Pr of the propellant. The valve units are then fitted to the rrlr~ t A; n Pr, Formulation 4 Tahlets each containing 60 mg of active ingredient are made as follows:
2 -Aminobicyclo[3.1.0]hexane-2,6-~;~Arh~xylic Acid 60 mg Starch 45 mg Microcrystalline ~Plll,loqe 35 mg Polyvinylpyrrolidone 4 mg Sodium carboxymethyl starch 4.5 mg - MAgnPq;nm stearate 0.5 mg Talc ~ m~
Total 150 mg 2~ 9577~
W096/04900 PCT~Sg~l03l7 The active ingredient, starch, and cellulose are passed through a No. 45 mesh U.S. sieve and mixed thoroughly. The solution of polyvinylpyrrolidone is mixed with the resultant powders which are then passed through a No. 14 mesh U.S. sieve. The granules so produced are dried at 50~C and passed through a No. 18 mesh U.s~ sieve. The sodium carboxymethyl starch, magnesium stearate, and talc, previously passed through a No. 60 mesh U.S. sieve, are then added to the granules which, after mixing, are compressed on a tablet machine to yield tablets each weighing 150 mg.
FormulatioA 5 Capsules each ~nr~ining 80 mg of active ingredient are made as follows:
2-~min~hi~yclo[3,1.0]hexane-2,6-dicarboxylic Acid 80 mg Starch 59 mg Microcrystalline cellulose 59 mg M~gn~qinm stearate 2 ma Total 200 mg The active_ingredient, cellulose, starch, and magnesium stearate are blended, passed through a No. 45 sieve, and filled into hard gelatin capsules in 200 mg quantities.
Formulation 6 Suppositories each c~nr~;n;ng 225 mg of active ingredient may be made as follows:
2-Pmin~h;cyclo[3.1.0]hexane-2,6-dicarboxylic Acid 225 mg Saturated fatty acid glycerides2.000 ma Total 2,225 mg ~ W096/04900 2 ~ q ~ 7 7 ~ r~
The active ingredient is passed through a No. 60 mesh U.S. sieve and suspended in the saturated fatty acid glycerides previously melted using the minimum heat n~cecq~ry. The mixture is then poured into a Suppository mold of nominal 2 g capacity and allowed to cool.
Formulation 7 Suspensions each ~nnt~;ning 50 mg of active ingredient per 5 ml dose are made as follows:
2-Aminobicyclo[3.1.0]hexane-2,6-dicarboxylic Acid 50 mg Sodium carboxymethyl cellulose 50 mg Syrup 1.25 ml senzoic acid solution 0.10 ml Flavor q.v.
Color q.v.
Purified water to total 5 ml The ~~l;~ nt iS passed through a No. 45 mesh U.S.
sieve and mixed with the sodium carboxymethyl cellulose and syrup to form a smooth paste. The benzoic acid solution, flavor and color are diluted with some of the water and added, with stirring. Sufficient water is then added to produce the required volume.
Formulation 8 An inLldv~llols f~ l~r;on may be prepared as follows:
2-P~~;nnbi-~yclo[3.1.0]hexane-2,6-dicarboxylic Acid 100 mg Mannitol 100 mg 5 N Sodium hydroxide 200 ml Purified water to total 5 ml The following ~ _lPq further illustrate methods for their synthesis of (+)-2-aminobicyclo[3.1.0]hexane-. _ .. .. .... . .. . _ _ _ _ _ _ . . .
~1 95779 Wo96/04soo ~I/U~,3~ 7 -2,6-dicarboxylic acid. The Examples are not intended to be limiting to the scope of the invention in any respect, and should not be so construed All experiments were run under a positive pressure of dry nitrogen or argon. All solvents and reagents were purchased from commercial sources and used as received, unless otherwise indicated. Dry tetrahydrofuran (THF) was rhti~in Pd by disti~ ti~n from sodium or sodium benzophenone ketyl prior to use. Proton nuclear magnetic resonance tl~ N~R) spectra were nhtilirrd on a GE QE-300 spectrometer at 300.15 M~z, a Bruker AM-500 i~e~LL~ ?trr at 500 M~z, or a Bruker AC-200P
spe~LL, -trr at 200 M~z. Free atom bombardment mass spectroscopy (FAsMS) was performed on a VG ZAB-2SE
instrume t. Field desorption mass spectroscopy tFDMS) was performed using either a VG 70SE or a Varian MAT 731 in~LLI . Optical rotations were measured with a Perkin-Elmer 2G1 polarimeter. Chromatographic separation on a Waters Prep 500 LC was generally carried out using a linear gradient o~ the solvents in~ t~ in the text. The reactions were generally monitored for completion using thin layer chromatography (~LC~. Thin layer chromatography was performed using E. Merck Kieselgel 60 F254 plates, 5 cm x ~LO cm, 0.25 mm thirkn~c8 Spots were detected using a , ,ini~tir,n of W and chemical detection (plates dipped in a ceric i ;nm molybdate solution [75 g of i inm molybdate and 4 g of cerium (IV~ sulfate in 500 mL of 10% aqueous sulfuric acid] and then heated on a hot plate). Flash chromatography was performed as described by Still, et al. Still, Kahn, and Mitra, J. Org. C~em., 43, 2923 (1978). ~ l analyses for carbon, hydrogen, and nitrogen were ~=rtrrm;ned on a Control F~li, ' Corporation 440 r,l. ti~l Analyzer, or were performed by the Universidad Complutense Analytical Centre (Facultad de Farmacia, Madrid, Spain). Melting points were ~etQrmin~d in open glass capillaries on a Gallenkamp hot .. , ,, .. . , . , .. ... ... .. .. . _ . . . .. . . _ . _ _ 21 9577~
096/04900 PCT~S95/10317 air bath melting point apparatus or a shchi melting point apparatus, and are uncorrected.
Preparation 5~ArhPt~n~ymethyl Dimethylsulfonium sromide A solution of ethyl bromoacetate (265g) and dimethyl sulfide (114g) in acetone ~500m3) was stirred at room temperature. ~fter three days, the title ~ ~ ~ was isolated by filtration of the reaction mixture. Melting point 88-90~C.
3xample (lSR,5Rs,6SR) Ethyl 2-Oxobicyclo[3.1.0]h OE ane-6-15carboxylate A suspension of carbethoxymethyl diethylsulfonium bromide (~5.5g) in toluene (350mL) was treated with 1,8-~;A7~hi~yclo[5.4.0]undec-7-ene (30.2g). The resulting mixture was stirred at room temperature. After one hour, the reaction mixture was treated with 2-cyclopenten-l-one (19.57g). After an additional 18 hours, the reaction mixture was added to a 1 N
hydrochloric acid/sodium chloride solution. The resulting mixture was OE tracted with diethyl ether. The ~ ;nP~ ether extracts were dried over r-gn~q;~lm sulfate, filtered, and ~nn~Pntrated in vacuo. The residue was purified using silica-gel ~l1L~ tn~raPhY, eluting with a linear gradient of 10% ethyl acetate/hexanes to 50% ethyl acetate/h OE anes, to give 22.81g of the title compound. Melting point: 36-38 C.
FDMS: m/z = 168 (M+).
Analysis calculated for CgH12O3: C, 64.27; H, 7.19.
Found: C, 64.54; H, 7.11.
7 7 ~
W096!04900 ~CTNS95/l0317 Exampl e 2 (lSR,2RS,SRs,65R) Diethyl 2-P~;nnhiryclo[3.1.0]-hexane-2,6-dicarboxylate and (lSR,25R,5RS,6SR) Diethyl 2-~minnh;ryclo[3.1.0]-hexane-2,6-~;rArhn~ylate A solution of the compound prepared as described in Example 1 (22. 81g) in ethanol (200mL) was treated with an aqueous solution of potassium cyanide (9.71g) and i1lm carbonate (21.2g) in water (200mL). The resulting mixture was heated to about 50 C. After about 18 hours, the reaction mixture was allowed to cool to room temperature and treated with sodium hydroxide (16.2g). The resulting mixture was heated to reflux.
After about 18 hours, the reaction mixture was allowed to cool to room temperature, then cooled to O C. The pH
of the cold mixture was adjusted to pH 1 by the A~;t;nn of concentrated hydrochloric acid. This mixture was concentrated to dryness in vacuo. The residue was dissolved in ethanol, cooled to O C, and treated with thionyl chloride~(80.6g). ~he resulting mixture was heated to reflux. After about 48 hours, the reaction was concentrated to dryness i~ vacuo. ~he residue was 25 treated with 1 N sodium hydroxide, and the resulting mixture extracted with diethyl ether. The - ' in~d ether extracts were dried over potassium rArhnnAre, filtered, and concentrated in vac~o to give 24.6g of a mixture of the title compounds.
~xample 3 (lSR, 2SR, 5RS, 6SR) Diethyl 2-Aminobicyclo[3~l~o]
hexane-2,6-~;cArbn~ylate A solution of the _ ~c prepared as described in Example 2 (20.71g) in ethyl acetate (200mL) was treated with a solution of oxalic acid ~15.46g) in ~ WO9~/04900 2 1 q57 7~ PCT~S95/10317 ethanol (5omr~). The resulting mixture was stirred at room temperature. After one hour, the reaction mixture was treated with additional ethanol (50m~). After 18 hours, the mixture was filtered, and the filtrate was evaporated to dryness in vacuo. The residue was treated with 1 N sodium hydroxide, and the resulting mixture extracted with diethyl ether. The , ';nP~ ether extracts were washed with brine, dried over potassium carbonate, filtered, and concentrated in vacuo. The residue was purified by silica-gel chromatography, eluting with methylene chloride:5% i inm hydroxide/--~hi~nnl (97:3), to give 15.41g of the title compound.
PDMS: m/z = 242 (M+H).
Analysis calculated for C12Hl9N~4: C, 59.74; H~ 7.94;
N, 5.81. Found: C, 59.78; H, 8.13; N, 5.77.
Exam~le 4 20(-)-Diethyl 2-~m; nnh; ~yclo[3.1.0]hexane-2,6-dicarboxylate ~ solution of the racemic mixture of compounds prepared as described in Example 3 (6.56g) in ethyl acetate (lOOmL~ was treated with a solution of (+~-di-p-toluoyl-D-tartaric acid (12.0g) in ethyl acetate (loomL). After standing overnight at room temperature, the crystalline solid was removed by filtration and dried to give 14.7g. Additional crystalline solid was obtained by cooling the filtrate to O C. The combined crystalline solids were dissolved in hot ethyl acetate, cnntiq;n;n~ enough 2- propanol for complete dissolution.
~After cooling to O C, the crystalline solid was isolated by filtration, to give 2.3g of a solid having an ~ni~nt; iC excess of ~ 95%. The freebase form was oht~;no~ by partitioning the salt between a~ueous sodium h;~arh~ni~te and ethyl acetate. The organic phase was 2~ q~77~
w096/04900 PCT~59~10317 separated, dried over potassium carbonate, filtered, and c~nrpntrRtp~ in vaCuo to give .77g of the title compound.
EDMS: m/z = 242 (M+H).
Optical rotation: aD = -5.15- (c = 1, EtOH).
Analysis calculated for C12H19N~4: C~ 59-74; H~ 7-g4;
N, 5.81. Found: C, 59.68; H, 8.13; N, 5.58.
~xample 5 (+)-2-~minnhicyclo[3.1.0]hexane-2,6-dicarboxylic Acid A solution of the compound prepared as described in Example 4 (0.69g~ in tetrahydrofuran (lOm~) was treated with 1 N sodium hydroxide (lOm~1, and the resulting mixture vigorously stirred at room te-m~perature~ After several days, the title compound was i crl ~tP~ by anion-exchange chromatography (sio-Rad AG1-X8), eluting with 50% acetic acid/water, to give 0.53g of the title 1 _ln~
FDMS: m/z = 186 tM+H).
Optical rotation: aD = 21.32- (c = 1, 1 N HC1).
Analysis calculated for CgH11N04-1.25H20: C, 46.26; E~, 6.55; N, 6.74. Found: C, 46.68; ~, 6.47; N, 6.49.
Example 6 2-Oxobicyclor3.1.0]hexane-6-carboxylic acid A mixture of 60~g of (lSR,5RS,6SR) ethyl 2-oxobicyclo[3.1.0]
hexane-6-carboxylate and 300 ml of 1 N sodium hydroxide was stirred at 25-30~C. After 2.5 hours, concentrated hydrochloric acid was added to adjust the pH to 0.8-1.2. The resulting solution was extracted with ethyl acetate. The extracts were dried over r~gnPq;llm sulfate, filtered, and concentrated to give 49.1 g (98%) of the crude material.
~ W096/04900 2 1 9 5 7 7 9 pCT~S9~10317 RecrysrAl1;~atinn from 100 ml of ethyl acetate ~ave the title c ~, mp 123.5-128~C.
S: m/z = 140 (M+) Analysis calculated for C7HgO3 C, 60.00; H, 5.75. Found:
C, 60.14; H, 5.79.
Example 7 2-Oxobicyclo[3.1.0]hexane-6-carboxylic acid salt with (S)-1-phenylethylamine A solution of 14 g of the compound prepared in Example 6 in 140 ml of 25% ethanol in ethyl acetate was ~ -in~ with (S)-1-phenylethylamine (1 eq.l. After stirring overnight, the precipitated salt was isolated by filtration and dried to give 11.87 g (45.4%) of the desired salt. Conversion of the salt to the partially resolved 2-oxobicyclo[3.1.0]hexane-6-carboxylic acid by the method of Example 8 and analysis indicated that the salt was 68% ee. The ~n~nti~ iC excess was determined by conversion to the methyl ester with diazomethane followed by chiral ~PLC on a Chiralpak AS column at 40~C eluted with 10% isopropanol/90% hexane at 1 ml/min with detection at 210 nm.
Exam~le 8 (+)-2-Oxobicyclo[3.1.0]hexane-6-carboxylic acid A mixture of 1.31 g of the product of Example 7 and 10 ml of lN hydrochloric acid was stirred for 5 minutes and extracted with ethyl acetate. The extracts were dried over sodium sulfate, filtered, and concentrated to give 0.61 g of the title compound, mp 110-115~C. The product was detPrmi n~d to be 68% ee by chiral HPLC ~method of Example 7).
FDMS: m/z = 141 (M+H) Optical Rotation: ~ D = 49.85~
21 ~5779 W096/04900 ~ ,J/l0 Fxam~le 9 ~-)-2-Spiro-5'-hy~Antn;nh;cyclo[3.1.0]hexane-6-carboxylic acid A solution of the ~ dL ~d as described in Example 8 ~68~ ee, 1 eq.), potasgium cyanide ~1.25 eg.), and , ;nm ri~rhnnAte ~2.5 eq) were combined and stirred in ethanol/water at 25~C for 40 hours. The mixture was Ac;~;f;~ with 6N
hydrochloric acid, ~nnr~ntrated, diluted with water, and filtered to give a 79% yield of a 90:10 mixture of diastereomers, mp 286-290~C. ~he diastereomeric mixture was recrystAl1;7P~ ~rom isopropanol/water to give in 48% yield the title ~- ~ in 100% diastereomeric and 100%
enantiomeric purity (~nAnt;~ ic ratio ~t~rm;ne~ by chiral HPLC on a 4.6 x 150 mm Chiralcel OD-H column, eluted with 15 ~ isopropanol/85% hexane at 1 ml/min at 40~C with detection at 220 nm; diastereomeric ration determined by HPLC on a Zorbax ss-phenyl column at 40~C with elution with 90:10 buffer~acetonitrile eluted at 2 ml/min with ~t~t;nn at 220 nm ~buffer = 0_1 ~ dibasic sodium phns~hAt~ monohydrate adjusted to pH 2 1 with phosphoric acid).
FDMS: m/z = 211 ~M + H) Optical Rotation: a D = - 25.98~
Analysis calculated for C9HlON204: C, 51.43: H, 4.79; N, 13.33. Found: C, 51.38; H, 4.80; N, 13.26.
~xample 10 Ethyl 2-spiro-5l-hydantoinbicyclo[3~l~o]hexane-6-carboxylate A mixture of 5.~5 g of ethyl 2-oxobicyclo[3.1.0]hexane-6-carboxylate, 2.15 g of potassium cyanide, 5.77 g of i ;~lm ~ArhnnAte~ 30 ml of 2s-3 ethanol, and 12 ml of water was stirred at 35~C until the reaction was complete hy HPLC.
After 15 hours, the reaction mixture was cooled to 0~C and 33 ~ W096/04900 219~7~9 ~"~ r~l7 ml of-water was added to the mixture. After 2 hours at 0~C, the precipitate was isolated by filtration and dried to give 5.23 g (73~) of the title compound, mp 217-220~C.
S FDMS: m/z = 238.1 (~+) ~ Analytical r~1r~ tPd for C11H14N2O4: C, 55.46; H, 5.92;
N, 11.76. Found: C, 55.74; H, 5.88; N, 11.50.
Exam~le 11 2-Spiro-5'-hy~nto;rhicyclo[3.1.0]hexane-6-carboxylic acid A mixture of 16.32 g of the product of Example 10 and 137 ml of 2N NaOH was stirred at 25~C. After 1 hour, concentrated hydrochloric acid was added to adjust the pH to 1Ø The resultiny precipitate was isolated by filtration and dried to give 13.70 g (95%) of the title compound, mp 277-279~C.
FD~S: m/z = 210.1 ~+) Analysis r~lcl~latPd for C9~10N2~4: C, 51.43; H, 4.79; N, 13.33. Found: C, 51.70; ~, 4.93; N, 13.43.
Exam~le 12 2-Spiro-5~-hy~nto;nh;cyclo[3.1.0]hexane-6-carboxylic acid, (S)-l-phenyle t hylamine salt A mixture of 1.05 g of the product of Example 11 and 16.6 ml of a 1.6 : 1 solution of acetone : water was stirred at 25~C
while adding 1.53 g of R-(+)-1-phenylethylamine. The mixture was stirred for 2 hours at room temperature. The crystals were filtered, rinsed with acetone, and dried to give 0.74 g (45~) of the title compound, mp 205-212~C.
Optical Rotation: ~ D = -31.88~ (c = 1, methanol) 21 ~779 W096/04900 pcTNss~llo3l7 -~xa~ple 13 ~ 2-Spiro-5 ' -hy~An t n i nh; cyclo[3.1.0]hexane-6-carboxylic acid A mixture of 0.74 g of the product of Example 12 and 10 ml of water was stirred at 25~C while the pH was adjusted from 6.81 to 1.0 using lN HCl. The reaction mixture was stirred for 1 hour and the product was rgll ~rt~d by filtration and dried to give 0.35 g ~75~ of the til-le , __ ~, mp 310~C (decomp).
FDMS: 210.1 (M+) Optical Rotation: a D = -24.22~ (c = 1, methanol) Analysis calculated for CgHloN2O4: C, 51.43: H, 4.80;
N,13.33. Found: C, 51.67; H, 4.87; N, 13.61.
Rxa~le 74 (+)-2-Aminobicyclo[3.1.0]hexane-2,6-dicarboxylic Acid A solution of 184 g of (-)-2-spiro-5~-hydantoinbicyclo[3.1.0]
hexane-6-carboxylic acid and 1750 ml of 3N NaOH was heated at reflux until the reaction was complete by HPLC. After 28 hours, the solution was cgoled to room temperature and filtered through glass paper to remove trace amounts of insoluble material. The p~ of the sslnt;nn was adjusted to 3.0 using concentrated ~Cl. ~he reaction mixture was stirred 1 hour at room temperature and two hours at 0~C. The precipitated product was collected by filtration, washed with 170 ml of cold water ar,d dried to give 152.5 grams (86~) of the title compound.
FDMS: m/z = 186.1 (~+1) optical rotation: a D = 23.18~ (c = 1, lN ~Cl)
It has now been found that a , _ fl which is an agonist that acts selectively at negativelY coupled cAME-linked rihntropic glutamate receptors is capable of reducing startle response in rats following the cessation of chronic nicotire or fliA7ep-~m exposure. It is believed that this finding portends that any agonist that acts at negatively coupled cAMP-linked metabotropic glutamate receptors will be useful for treating substance withdrawal, and indeed may be useful for providing protection against substance dependence.
According-to one aspect, therefore, the present invention provides the use of an agonist which acts at negatively coupled cANP-linked metabotropic glutamate receptors for protecting a warm blooded ma~mal from dependence on a-substance.
It has ~een found that a novel compound which is an agonist that acts selectively at negatively coupled cAMP-linked metabotropic glutamate receptors, ~+)-2-aminobicyclo[3.1.0]hexane-2,6-dicarboxylic acid is effective in the rat startle models for nicotine and diazepam withdrawal. Accordingly it is believed that any compound which acts as an agonist at negatively coupled cAMP-linked metabotropic glutamate raceptors, especially any agonist which acts selectively, will be useful for the treatment of the withdrawal or CPqsati nn of these and other dependence-producing substances. Furthermore, since withdrawal isinrir~tPly related to flPpPnflPn~e and to tolerance, it is believed that an agonist which acts at negatively coupled cAME~-linked metabotropic glutamate receptors will also be useful for the treatment of substance dependence and tolerance, and indeed generally to protect a warm blooded mammal from flPpPnflPn~e on a flPpPnflPnnP-producing substance.
~ W096/04900 2 1 9 5 7 7 q PCT~S9~10317 The ~PpPn~Pnre-producing substance may be, for example, an opiate, benzodiazepine, nicotine, cocaine or ethanol.
According to another aspect, the present invention provides the use of an agonist which acts at negatively coupled cAMP-linked metabotropic glutamate receptors for the treatment of drug tolerance, withdrawal or cessation.
According to yet another aspect, the present invention provides the use of an agonist which acts at negatively coupled cA~P-linked metabotropic glutamate receptors for the treatment of smoking cPq~ti nn .
The particular dose of agonist administered will of course be ~PtPrminpd by the particular circumstances surrounding the case, ;nrln~inr the activity of the particular agonist administered, the route of administration, the particular condition being treated, and similar considerations. The agonist can be administered by a variety of routes inr1ll~ing oral, rectal, trAnc~Prr-l, subcutaneous, intravenous, intramuscular, or intranasal routes. Alternatively, the agonist may be administered by continuous infusion. A
typical daily dose will contain from about O.OOl mg/kg to about lO0 mg/kg of the agonist. Preferably, daily doses will be about 0.05 mg/kg to about 50 mg/kg, more preferably from about O.l mg/kg to about 20 mg/kg.
According to preferrea aspect, the present invention provides the use of an agonist as defined hereinabove for use in a method of protecting a warm-blooded mammal from ~PpPn~Pnre on a dependence-producing pharmaceutical, for example, a bPn7o~iA7~r;np such as ~;~7Pp~m. In this method, the agonist may be administered before said phArr--Putical is first administered, after said phArr--Plltir~l has been administered or after said phArr-rPnt;rAl has been withdrawn, or it may be co-administered with said pharmaceutical.
Agonists which act at negatively coupled cANP-linked metabotropic glutamate receptors may be PCTIUS 9 5 / 1~ 3 ~-g329D ~ 9~7~q IPEAIUS lb AUG 1996 identified using the following ~Yp~ri L. Firstly, the affinity of a test compound for metabotropic glutamate receptors may be demonstrated by the selective disrlA~ L of (15,3R)-1-aminocyclopentane-1,3-~io~rh~ylic acid-sensitive [3H]glutamate binding to rat brain cell membranes. The binding of [3H]glutamate ([3H]Glu) is conducted with crude membranes of rat forebrain as described by Schoepp and True. Schoepp and True, Neuroscience Lett., 145, 100-104 (1992); Wright, McDonald, and Schoepp, J. Neurochem., 63, 938-945 (1994). The affinity of a test ~ _od for the receptor may be expressed as the concentration of the test compound that inhibits 50i binding (ICso), or the percent dis~l~, L of [3H]Glu at a 10 yM or 100 yM
concentration of the formula I c ~. In this test, the ICso for (+)-2-aminobicyclo[3.1.0]hexane-2,6-~;o~rh~xylic acid was found to be 0.18 yM.
The ability of a test . _nd to act as an agonist at negatively coupled cAMP-linked metabotropic receptors may be measured using the following method. Test c, ullds are tested for their ability to decrease forskolin-stimulated cAMP formation in the rat h;rporP ,us and the rat cerebral cortex, using the ~-~cedul~s described in Schoepp and Johnson. Schoepp and Johnson, Neurochem. Int., 22, 277-283 (1993). In this test, (+)-2-~min~h;~yclo[3~l.o]hexane-2~6-~ie~rh~ylic acid was found to give the result shown in Table II below.
Table II. Inhibition of Forskolin-6timulated cAMP
F. i~
ECso (yM) Rat cerebral cortex .055 + .017 Rat hi,p~- ~ .036 1 .015 -9329D ~ ~ 1996 21 ~5779 The ability of negatively coupled cAMP-linked metabotropic receptor agonists to protect a warm blooded ma_mal from the effects of drug withdrawal or cessation may be demonstrated using an auditory startle model. In 5 this model, animals are dosed with a drug (nicotine or ~;?70p~m), then dosing is discontinued. This cessation of drug dosing elicits an increased startle response to auditory stimuli. Test compounds are then administered to animals to determine whether they are capable of 10 attenuating the increased startle response.
Long Evans rats t200-400 g; Harlan Sprague Dawley, Columbus, IN) were individually housed in a controlled environment on a 12 hour light-dark cycle and given free access to food (Purina Rodent Chow) and water. Rats 15 were anesthot;7ed with isoflurane and Alzet osmotic pumps (Alza Corporation) were implanted su~cutaneously.
Test c- 1 was dissolved in a vehicle of purified water and neutralized with 5N NaOH to a pH of 7-8 when appl;~hlo. Diazepam (Sigma Chom;~l Company, 20 St. ~ouis, MO) was su~pon~od in a vehicle consisting of 40% PEG 300, 10% EtOH, 2% benzyl alcohol, 1% Tween 80, and 47% purified water. Nicotine (Research Pio~hom;~
Inc., Natick, MA) was dissolved in saline. Control animals received the respective vehicle.
25 Nicotine withdrawal: Pumps were filled to deliver nicotine (6mg/kg/day s.c.), diazepam (lOmg/kg/day s.c.), test compound (0,1,3,1Omg/kg s.c.) or vehicle. Twelve days following subcutaneous implantation of pumps, rats were anesthetized with isoflurane and the pumps were 30 removed. During withdrawal (following pump removal), the auditory startle response (peak amplitude, Vmax) of individual rats was recorded using San Diego Instruments startle rhl ~ (San Diego, CA). Startle sessions consisted of a 5-minute adaptation period at a 35 background noise level of 70+2dBA immediately followed by 25 presentations of auditory stimuli (120+2d~A noise, 50ms duration) presented at 8-second intervals. Peak W096l04900 21 9577~ PCT~S9S/10317 -startle amplitudes were then averaged for~all 25 presPnt~t; nnc of stimuli for each session and all data are preseAted here as overall session means. Auditory startle rPcpnn~; ng was evaluated daily on withdrawal days 1,2,3,4 or 5. saseline startle rpcpnn~ing was evaluated prior to pump removal on day 12.
Auditory startle rPcpnn~ing was significantly increased through the first three days following cPcqstinn of chronic nicotine exposure when compared to control rats receiving water. Rats given a rep~
dose of nicotine at doses of 0.03 mg/kg, i.p. or higher did not display the same heightened startle response seen for aAimals with no nicotine rPpl~n~ ~nt.
Pretreatment with ~+)-2-aminobicyclo[3.1.0]hexane-2,6-dicarboxylic acid produced a dose-dependent hlonk~P Of the withdrawal-induced increase in startle responding as well. A significant ~ttPnn~tinn of the heightened startle was apparent at 3mg/kg, p.o. dose of the compound when compared to nicotine coAErols (ED50=0-7 mg/kg i.p.).
D~7f~m Withdrawal: Auditory startle r~Qpnn~ing was qi~ni~in~ntly increased through the first four days following cessation of chronic dia_epam exposure when compared to control rats receiving vehicle Replacement doses of 3 and lO mg/kg, i.p. ~;~7ap~m did not block the increased start~le rPqpnn~;ng and in some ;nqt~nnPR
further iAcreased reactivity indicating toleraAce. Rats which received 30 mg/kg, i.p. ~;~7Pp~m rep~ daily 60 minutes before ev~ln~tinn of startle roqpnn~;ng, did not show increased reactivity following ~;~7Pp~m C~qq~ti nn on days 1 through ~ when I ed to the ~i~7Pp~m control. Pretreatment with (+)-2-innhinyc1O[3~l~o] hexane-2,6-dicarboxylic acid blocked the expected increase in startle rpcpnn~i ng which followed cessation of diazepam exposure. 3Oses of 0.1 aAd 0.3 mg/kg, p.o. of the c _ ~ significaAtly ~ W096/04900 2 1 9 5 7 7 9 PCT~S95/10317 ~ttonn~ted enhanced startle when compared to control rPcprm~;n!J ~ED50=0.1 mg/kg, p.o. ) .
(+)-2-Aminobicyclo[3.1.0]hexane-2,6-dicarboxylic acid may be prepared by reacting carbethoxYmethyl dimethy1c-~1f~nillm bromide with 2-cyclopenten-1-one in the presence of a base, such as 1,8-diazabicyclo[5.4.0]undec-7-ene to afford ethyl 2-oxobicyclo[3.l~o]hexane-6-carboxylate. This ester may then be reacted with an aqueous solution of potassium cyanide or sodium cyanide and . illm carbonate to produce an int~L -~;Ate hydantoin, (the sucherer-sergs reaction), which is then hydrolysed using sodium hydroxide, to afford a diastereomeric mixture of diethyl 2-aminobicyclo[3.1.0]hexane-2,6-~;c~rh~ylates. The desired diastereomer may be obtained by crystallization wi'th oxalic acid. This diastereomer may then be resolved by forming a crystalline salt with (+)-di-p-toluoyl-D-tartaric acid and recovering (-)-diethyl Z-aminobicyclo[3.1.0]hexane-2,6-dicarboxylate. Xydrolysis of this diester using aqueous sodium hydroxide gives (+)-2-aminobicyclo[3.1.0]hexane-2,6-dicarboxylic acid.
Alternatively, the ethyl 2-oxobicyclo[3.1.0]hexane-6-carboxylic acid may be hydrolysed using sodium hydroxide to give 2-oxobicyclo[3.1.0]hexane-6-carboxylic acid.
This acid may then be resolved by formin~ a crystalline salt with (S)-1-phenylethylamine and recovering (+)-2-oxobicyclo[3.1.0]hexane-6-carboxylic acid. This acid may then be converted into (+)-2-aminobicyclo[3.1.0]
hexane-2,6-dicarboxylic acid by reaction with an aqueous solution of potassium cyanide or sodium cyanide and ammonium carbonate to produce an intermediate hydantoin (the sucherer-sergs reaction) followed by hydrolysis of the hydantoin using sodium hydroxide. This procedure may also be modified by performing the resolution step on the hydantoin rather than on the 2-oxobicyclo[3.1.0]hexane-6-carboxylic acid. In this W096/04900 2 1 9 5 7 7 ~ PCT~S9~10317 -case, (~ phenylethylamine has been found to be a suitable resolving agent.
The agonists are preferably formulated prior to administration in , ~ in~t;nn with one or more pharm--Pllt;r~lly-acceptable carriers, ~ lPntq, or ~ripi~ntq. The pharmaceutical formulations are prepared by known procedures using well-known and readily availa~le ingredients. In making the compositions, the active ingredient will usually be mixed with a carrier, or diluted by a carrier, or enclosed within a carrier, and may be in the form of a capsule, sachet, paper, or other nnnt~inPr, When the carrier serves as a diluent, it may be a solid, semi-solid, or liriuid material which acts as a 'vehicle, ~nipi~nt, or medium for t-h-e active ingredient. The compositions can be in the form of tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols, ni ' c cnnt~;ning, for example, up to 10% by weight of active compound, so~t and hard gelatin capsules, suppositories, sterile iniectable Snl--tinnq, dermal patch, snhcllt~n~ous implant, and sterile packaged powders.
Some examples o~ snitRhl~ carriers, excipients, and diluents include lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum, acacia, calcium phnsph~te~
alginates, trAgar~nth, gelatin, calcium silicate, microcrystalline cel1lllose, polyvinylpyrrolidone, cellulose, water syrup, methyl cellulose, methyl and propyl hydroxyhPn7oat~q, talc, r~gn~qinm stearate, stearic acid, and mineral oil. The f~ lat;nnq can additionally include lubricating agents, wetting agents (surfactants), emulsifying and suspending agents, preserving agents, sweetening agents, or flavoring agents. Compositions may be f~ l~t~ 50 as to provide r~uick, sustained, or delayed release of the active ~ W096/~900 2 ~ ~5779 P~ 17 ingredient after administration to the patient by employing procedures well known in the art.
The compositions are preferably formulated in a unit dosage form, each dosage crntAininr from about 1 mg to about 500 mg, more preferably about 5 mg to about 200 mg of the active ingredient. The term ~unit dosage form'~ refers to a physically discrete unit suitable as unitary dosages for human subjects and other mammals, each unit rnntA;ninr a predetrrmin~d quantity of active material calculated to produce the desired therapeutic effect, in association with a suitable ph~rrace1ltir~
carrier, diluent, or excipient. The following formulation examples are illustrative only and are not --int~n~ to limit the scope of the invention in any way.
2~ 95779 W096/04900 PCT~S95/10317 -Formulation ~ard gelatin capsules are prepared using the ~ollowing ingredients:
Quantity (mg/capsule) 2-Aminobicyclo[3.1.0]hexane-2,6-dicarboxylic Acid 250 Starch, dried 200 ~agnesium stearate 10 Total 460 mg 20The above ingredients are mixed and filled into hard gelatin capsules in 460 mg quantities.
Formulation 2 A tablet is prepared using the ingredients below:
ouantity 30~mg/tablet) 2-~mi nnh; cyclo[3.1.0]hexane-2,6-dicarboxylic Acid 250 35 rPl l n l nse, microcrystalline400 Silicon dioxide, fumed 10 Stearic acid . 5 Total 665 mg The c ~ Pntc are blended and compressed to form tablets each weighing 665 mg.
~ W096104900 2 1 9 ~ 7 7 9 PCT~S95/10317 Formulation 3 An aerosol solution is prepared ~ntAining the following , ~ntc:
. .
Weight 2-Aminobicyclo[3.1.0]hexane-2,6-~iCArh~xylic Acid 0.25 Ethanol 29 . 75 Propellant 22 70 . 00 ~chlorodifluoromethane) Total 100.00 The active compound is mixed with ethanol and the mixture added to a portion of the PropPl 1 Anr 22, cooled to -30~C and transferred to a filling device. The re~uired amount is then fed to a stainless steel r~ntAinPr and diluted with the 1 ;n~Pr of the propellant. The valve units are then fitted to the rrlr~ t A; n Pr, Formulation 4 Tahlets each containing 60 mg of active ingredient are made as follows:
2 -Aminobicyclo[3.1.0]hexane-2,6-~;~Arh~xylic Acid 60 mg Starch 45 mg Microcrystalline ~Plll,loqe 35 mg Polyvinylpyrrolidone 4 mg Sodium carboxymethyl starch 4.5 mg - MAgnPq;nm stearate 0.5 mg Talc ~ m~
Total 150 mg 2~ 9577~
W096/04900 PCT~Sg~l03l7 The active ingredient, starch, and cellulose are passed through a No. 45 mesh U.S. sieve and mixed thoroughly. The solution of polyvinylpyrrolidone is mixed with the resultant powders which are then passed through a No. 14 mesh U.S. sieve. The granules so produced are dried at 50~C and passed through a No. 18 mesh U.s~ sieve. The sodium carboxymethyl starch, magnesium stearate, and talc, previously passed through a No. 60 mesh U.S. sieve, are then added to the granules which, after mixing, are compressed on a tablet machine to yield tablets each weighing 150 mg.
FormulatioA 5 Capsules each ~nr~ining 80 mg of active ingredient are made as follows:
2-~min~hi~yclo[3,1.0]hexane-2,6-dicarboxylic Acid 80 mg Starch 59 mg Microcrystalline cellulose 59 mg M~gn~qinm stearate 2 ma Total 200 mg The active_ingredient, cellulose, starch, and magnesium stearate are blended, passed through a No. 45 sieve, and filled into hard gelatin capsules in 200 mg quantities.
Formulation 6 Suppositories each c~nr~;n;ng 225 mg of active ingredient may be made as follows:
2-Pmin~h;cyclo[3.1.0]hexane-2,6-dicarboxylic Acid 225 mg Saturated fatty acid glycerides2.000 ma Total 2,225 mg ~ W096/04900 2 ~ q ~ 7 7 ~ r~
The active ingredient is passed through a No. 60 mesh U.S. sieve and suspended in the saturated fatty acid glycerides previously melted using the minimum heat n~cecq~ry. The mixture is then poured into a Suppository mold of nominal 2 g capacity and allowed to cool.
Formulation 7 Suspensions each ~nnt~;ning 50 mg of active ingredient per 5 ml dose are made as follows:
2-Aminobicyclo[3.1.0]hexane-2,6-dicarboxylic Acid 50 mg Sodium carboxymethyl cellulose 50 mg Syrup 1.25 ml senzoic acid solution 0.10 ml Flavor q.v.
Color q.v.
Purified water to total 5 ml The ~~l;~ nt iS passed through a No. 45 mesh U.S.
sieve and mixed with the sodium carboxymethyl cellulose and syrup to form a smooth paste. The benzoic acid solution, flavor and color are diluted with some of the water and added, with stirring. Sufficient water is then added to produce the required volume.
Formulation 8 An inLldv~llols f~ l~r;on may be prepared as follows:
2-P~~;nnbi-~yclo[3.1.0]hexane-2,6-dicarboxylic Acid 100 mg Mannitol 100 mg 5 N Sodium hydroxide 200 ml Purified water to total 5 ml The following ~ _lPq further illustrate methods for their synthesis of (+)-2-aminobicyclo[3.1.0]hexane-. _ .. .. .... . .. . _ _ _ _ _ _ . . .
~1 95779 Wo96/04soo ~I/U~,3~ 7 -2,6-dicarboxylic acid. The Examples are not intended to be limiting to the scope of the invention in any respect, and should not be so construed All experiments were run under a positive pressure of dry nitrogen or argon. All solvents and reagents were purchased from commercial sources and used as received, unless otherwise indicated. Dry tetrahydrofuran (THF) was rhti~in Pd by disti~ ti~n from sodium or sodium benzophenone ketyl prior to use. Proton nuclear magnetic resonance tl~ N~R) spectra were nhtilirrd on a GE QE-300 spectrometer at 300.15 M~z, a Bruker AM-500 i~e~LL~ ?trr at 500 M~z, or a Bruker AC-200P
spe~LL, -trr at 200 M~z. Free atom bombardment mass spectroscopy (FAsMS) was performed on a VG ZAB-2SE
instrume t. Field desorption mass spectroscopy tFDMS) was performed using either a VG 70SE or a Varian MAT 731 in~LLI . Optical rotations were measured with a Perkin-Elmer 2G1 polarimeter. Chromatographic separation on a Waters Prep 500 LC was generally carried out using a linear gradient o~ the solvents in~ t~ in the text. The reactions were generally monitored for completion using thin layer chromatography (~LC~. Thin layer chromatography was performed using E. Merck Kieselgel 60 F254 plates, 5 cm x ~LO cm, 0.25 mm thirkn~c8 Spots were detected using a , ,ini~tir,n of W and chemical detection (plates dipped in a ceric i ;nm molybdate solution [75 g of i inm molybdate and 4 g of cerium (IV~ sulfate in 500 mL of 10% aqueous sulfuric acid] and then heated on a hot plate). Flash chromatography was performed as described by Still, et al. Still, Kahn, and Mitra, J. Org. C~em., 43, 2923 (1978). ~ l analyses for carbon, hydrogen, and nitrogen were ~=rtrrm;ned on a Control F~li, ' Corporation 440 r,l. ti~l Analyzer, or were performed by the Universidad Complutense Analytical Centre (Facultad de Farmacia, Madrid, Spain). Melting points were ~etQrmin~d in open glass capillaries on a Gallenkamp hot .. , ,, .. . , . , .. ... ... .. .. . _ . . . .. . . _ . _ _ 21 9577~
096/04900 PCT~S95/10317 air bath melting point apparatus or a shchi melting point apparatus, and are uncorrected.
Preparation 5~ArhPt~n~ymethyl Dimethylsulfonium sromide A solution of ethyl bromoacetate (265g) and dimethyl sulfide (114g) in acetone ~500m3) was stirred at room temperature. ~fter three days, the title ~ ~ ~ was isolated by filtration of the reaction mixture. Melting point 88-90~C.
3xample (lSR,5Rs,6SR) Ethyl 2-Oxobicyclo[3.1.0]h OE ane-6-15carboxylate A suspension of carbethoxymethyl diethylsulfonium bromide (~5.5g) in toluene (350mL) was treated with 1,8-~;A7~hi~yclo[5.4.0]undec-7-ene (30.2g). The resulting mixture was stirred at room temperature. After one hour, the reaction mixture was treated with 2-cyclopenten-l-one (19.57g). After an additional 18 hours, the reaction mixture was added to a 1 N
hydrochloric acid/sodium chloride solution. The resulting mixture was OE tracted with diethyl ether. The ~ ;nP~ ether extracts were dried over r-gn~q;~lm sulfate, filtered, and ~nn~Pntrated in vacuo. The residue was purified using silica-gel ~l1L~ tn~raPhY, eluting with a linear gradient of 10% ethyl acetate/hexanes to 50% ethyl acetate/h OE anes, to give 22.81g of the title compound. Melting point: 36-38 C.
FDMS: m/z = 168 (M+).
Analysis calculated for CgH12O3: C, 64.27; H, 7.19.
Found: C, 64.54; H, 7.11.
7 7 ~
W096!04900 ~CTNS95/l0317 Exampl e 2 (lSR,2RS,SRs,65R) Diethyl 2-P~;nnhiryclo[3.1.0]-hexane-2,6-dicarboxylate and (lSR,25R,5RS,6SR) Diethyl 2-~minnh;ryclo[3.1.0]-hexane-2,6-~;rArhn~ylate A solution of the compound prepared as described in Example 1 (22. 81g) in ethanol (200mL) was treated with an aqueous solution of potassium cyanide (9.71g) and i1lm carbonate (21.2g) in water (200mL). The resulting mixture was heated to about 50 C. After about 18 hours, the reaction mixture was allowed to cool to room temperature and treated with sodium hydroxide (16.2g). The resulting mixture was heated to reflux.
After about 18 hours, the reaction mixture was allowed to cool to room temperature, then cooled to O C. The pH
of the cold mixture was adjusted to pH 1 by the A~;t;nn of concentrated hydrochloric acid. This mixture was concentrated to dryness in vacuo. The residue was dissolved in ethanol, cooled to O C, and treated with thionyl chloride~(80.6g). ~he resulting mixture was heated to reflux. After about 48 hours, the reaction was concentrated to dryness i~ vacuo. ~he residue was 25 treated with 1 N sodium hydroxide, and the resulting mixture extracted with diethyl ether. The - ' in~d ether extracts were dried over potassium rArhnnAre, filtered, and concentrated in vac~o to give 24.6g of a mixture of the title compounds.
~xample 3 (lSR, 2SR, 5RS, 6SR) Diethyl 2-Aminobicyclo[3~l~o]
hexane-2,6-~;cArbn~ylate A solution of the _ ~c prepared as described in Example 2 (20.71g) in ethyl acetate (200mL) was treated with a solution of oxalic acid ~15.46g) in ~ WO9~/04900 2 1 q57 7~ PCT~S95/10317 ethanol (5omr~). The resulting mixture was stirred at room temperature. After one hour, the reaction mixture was treated with additional ethanol (50m~). After 18 hours, the mixture was filtered, and the filtrate was evaporated to dryness in vacuo. The residue was treated with 1 N sodium hydroxide, and the resulting mixture extracted with diethyl ether. The , ';nP~ ether extracts were washed with brine, dried over potassium carbonate, filtered, and concentrated in vacuo. The residue was purified by silica-gel chromatography, eluting with methylene chloride:5% i inm hydroxide/--~hi~nnl (97:3), to give 15.41g of the title compound.
PDMS: m/z = 242 (M+H).
Analysis calculated for C12Hl9N~4: C, 59.74; H~ 7.94;
N, 5.81. Found: C, 59.78; H, 8.13; N, 5.77.
Exam~le 4 20(-)-Diethyl 2-~m; nnh; ~yclo[3.1.0]hexane-2,6-dicarboxylate ~ solution of the racemic mixture of compounds prepared as described in Example 3 (6.56g) in ethyl acetate (lOOmL~ was treated with a solution of (+~-di-p-toluoyl-D-tartaric acid (12.0g) in ethyl acetate (loomL). After standing overnight at room temperature, the crystalline solid was removed by filtration and dried to give 14.7g. Additional crystalline solid was obtained by cooling the filtrate to O C. The combined crystalline solids were dissolved in hot ethyl acetate, cnntiq;n;n~ enough 2- propanol for complete dissolution.
~After cooling to O C, the crystalline solid was isolated by filtration, to give 2.3g of a solid having an ~ni~nt; iC excess of ~ 95%. The freebase form was oht~;no~ by partitioning the salt between a~ueous sodium h;~arh~ni~te and ethyl acetate. The organic phase was 2~ q~77~
w096/04900 PCT~59~10317 separated, dried over potassium carbonate, filtered, and c~nrpntrRtp~ in vaCuo to give .77g of the title compound.
EDMS: m/z = 242 (M+H).
Optical rotation: aD = -5.15- (c = 1, EtOH).
Analysis calculated for C12H19N~4: C~ 59-74; H~ 7-g4;
N, 5.81. Found: C, 59.68; H, 8.13; N, 5.58.
~xample 5 (+)-2-~minnhicyclo[3.1.0]hexane-2,6-dicarboxylic Acid A solution of the compound prepared as described in Example 4 (0.69g~ in tetrahydrofuran (lOm~) was treated with 1 N sodium hydroxide (lOm~1, and the resulting mixture vigorously stirred at room te-m~perature~ After several days, the title compound was i crl ~tP~ by anion-exchange chromatography (sio-Rad AG1-X8), eluting with 50% acetic acid/water, to give 0.53g of the title 1 _ln~
FDMS: m/z = 186 tM+H).
Optical rotation: aD = 21.32- (c = 1, 1 N HC1).
Analysis calculated for CgH11N04-1.25H20: C, 46.26; E~, 6.55; N, 6.74. Found: C, 46.68; ~, 6.47; N, 6.49.
Example 6 2-Oxobicyclor3.1.0]hexane-6-carboxylic acid A mixture of 60~g of (lSR,5RS,6SR) ethyl 2-oxobicyclo[3.1.0]
hexane-6-carboxylate and 300 ml of 1 N sodium hydroxide was stirred at 25-30~C. After 2.5 hours, concentrated hydrochloric acid was added to adjust the pH to 0.8-1.2. The resulting solution was extracted with ethyl acetate. The extracts were dried over r~gnPq;llm sulfate, filtered, and concentrated to give 49.1 g (98%) of the crude material.
~ W096/04900 2 1 9 5 7 7 9 pCT~S9~10317 RecrysrAl1;~atinn from 100 ml of ethyl acetate ~ave the title c ~, mp 123.5-128~C.
S: m/z = 140 (M+) Analysis calculated for C7HgO3 C, 60.00; H, 5.75. Found:
C, 60.14; H, 5.79.
Example 7 2-Oxobicyclo[3.1.0]hexane-6-carboxylic acid salt with (S)-1-phenylethylamine A solution of 14 g of the compound prepared in Example 6 in 140 ml of 25% ethanol in ethyl acetate was ~ -in~ with (S)-1-phenylethylamine (1 eq.l. After stirring overnight, the precipitated salt was isolated by filtration and dried to give 11.87 g (45.4%) of the desired salt. Conversion of the salt to the partially resolved 2-oxobicyclo[3.1.0]hexane-6-carboxylic acid by the method of Example 8 and analysis indicated that the salt was 68% ee. The ~n~nti~ iC excess was determined by conversion to the methyl ester with diazomethane followed by chiral ~PLC on a Chiralpak AS column at 40~C eluted with 10% isopropanol/90% hexane at 1 ml/min with detection at 210 nm.
Exam~le 8 (+)-2-Oxobicyclo[3.1.0]hexane-6-carboxylic acid A mixture of 1.31 g of the product of Example 7 and 10 ml of lN hydrochloric acid was stirred for 5 minutes and extracted with ethyl acetate. The extracts were dried over sodium sulfate, filtered, and concentrated to give 0.61 g of the title compound, mp 110-115~C. The product was detPrmi n~d to be 68% ee by chiral HPLC ~method of Example 7).
FDMS: m/z = 141 (M+H) Optical Rotation: ~ D = 49.85~
21 ~5779 W096/04900 ~ ,J/l0 Fxam~le 9 ~-)-2-Spiro-5'-hy~Antn;nh;cyclo[3.1.0]hexane-6-carboxylic acid A solution of the ~ dL ~d as described in Example 8 ~68~ ee, 1 eq.), potasgium cyanide ~1.25 eg.), and , ;nm ri~rhnnAte ~2.5 eq) were combined and stirred in ethanol/water at 25~C for 40 hours. The mixture was Ac;~;f;~ with 6N
hydrochloric acid, ~nnr~ntrated, diluted with water, and filtered to give a 79% yield of a 90:10 mixture of diastereomers, mp 286-290~C. ~he diastereomeric mixture was recrystAl1;7P~ ~rom isopropanol/water to give in 48% yield the title ~- ~ in 100% diastereomeric and 100%
enantiomeric purity (~nAnt;~ ic ratio ~t~rm;ne~ by chiral HPLC on a 4.6 x 150 mm Chiralcel OD-H column, eluted with 15 ~ isopropanol/85% hexane at 1 ml/min at 40~C with detection at 220 nm; diastereomeric ration determined by HPLC on a Zorbax ss-phenyl column at 40~C with elution with 90:10 buffer~acetonitrile eluted at 2 ml/min with ~t~t;nn at 220 nm ~buffer = 0_1 ~ dibasic sodium phns~hAt~ monohydrate adjusted to pH 2 1 with phosphoric acid).
FDMS: m/z = 211 ~M + H) Optical Rotation: a D = - 25.98~
Analysis calculated for C9HlON204: C, 51.43: H, 4.79; N, 13.33. Found: C, 51.38; H, 4.80; N, 13.26.
~xample 10 Ethyl 2-spiro-5l-hydantoinbicyclo[3~l~o]hexane-6-carboxylate A mixture of 5.~5 g of ethyl 2-oxobicyclo[3.1.0]hexane-6-carboxylate, 2.15 g of potassium cyanide, 5.77 g of i ;~lm ~ArhnnAte~ 30 ml of 2s-3 ethanol, and 12 ml of water was stirred at 35~C until the reaction was complete hy HPLC.
After 15 hours, the reaction mixture was cooled to 0~C and 33 ~ W096/04900 219~7~9 ~"~ r~l7 ml of-water was added to the mixture. After 2 hours at 0~C, the precipitate was isolated by filtration and dried to give 5.23 g (73~) of the title compound, mp 217-220~C.
S FDMS: m/z = 238.1 (~+) ~ Analytical r~1r~ tPd for C11H14N2O4: C, 55.46; H, 5.92;
N, 11.76. Found: C, 55.74; H, 5.88; N, 11.50.
Exam~le 11 2-Spiro-5'-hy~nto;rhicyclo[3.1.0]hexane-6-carboxylic acid A mixture of 16.32 g of the product of Example 10 and 137 ml of 2N NaOH was stirred at 25~C. After 1 hour, concentrated hydrochloric acid was added to adjust the pH to 1Ø The resultiny precipitate was isolated by filtration and dried to give 13.70 g (95%) of the title compound, mp 277-279~C.
FD~S: m/z = 210.1 ~+) Analysis r~lcl~latPd for C9~10N2~4: C, 51.43; H, 4.79; N, 13.33. Found: C, 51.70; ~, 4.93; N, 13.43.
Exam~le 12 2-Spiro-5~-hy~nto;nh;cyclo[3.1.0]hexane-6-carboxylic acid, (S)-l-phenyle t hylamine salt A mixture of 1.05 g of the product of Example 11 and 16.6 ml of a 1.6 : 1 solution of acetone : water was stirred at 25~C
while adding 1.53 g of R-(+)-1-phenylethylamine. The mixture was stirred for 2 hours at room temperature. The crystals were filtered, rinsed with acetone, and dried to give 0.74 g (45~) of the title compound, mp 205-212~C.
Optical Rotation: ~ D = -31.88~ (c = 1, methanol) 21 ~779 W096/04900 pcTNss~llo3l7 -~xa~ple 13 ~ 2-Spiro-5 ' -hy~An t n i nh; cyclo[3.1.0]hexane-6-carboxylic acid A mixture of 0.74 g of the product of Example 12 and 10 ml of water was stirred at 25~C while the pH was adjusted from 6.81 to 1.0 using lN HCl. The reaction mixture was stirred for 1 hour and the product was rgll ~rt~d by filtration and dried to give 0.35 g ~75~ of the til-le , __ ~, mp 310~C (decomp).
FDMS: 210.1 (M+) Optical Rotation: a D = -24.22~ (c = 1, methanol) Analysis calculated for CgHloN2O4: C, 51.43: H, 4.80;
N,13.33. Found: C, 51.67; H, 4.87; N, 13.61.
Rxa~le 74 (+)-2-Aminobicyclo[3.1.0]hexane-2,6-dicarboxylic Acid A solution of 184 g of (-)-2-spiro-5~-hydantoinbicyclo[3.1.0]
hexane-6-carboxylic acid and 1750 ml of 3N NaOH was heated at reflux until the reaction was complete by HPLC. After 28 hours, the solution was cgoled to room temperature and filtered through glass paper to remove trace amounts of insoluble material. The p~ of the sslnt;nn was adjusted to 3.0 using concentrated ~Cl. ~he reaction mixture was stirred 1 hour at room temperature and two hours at 0~C. The precipitated product was collected by filtration, washed with 170 ml of cold water ar,d dried to give 152.5 grams (86~) of the title compound.
FDMS: m/z = 186.1 (~+1) optical rotation: a D = 23.18~ (c = 1, lN ~Cl)
Claims (22)
1. The use of an agonist which acts at negatively coupled cAMP-linked metabotropic glutamate receptors for the manufacture of a medicament for protecting a warm blooded mammal from dependence on a substance.
2. Use as claimed in Claim 1, in which said substance is an opiate, benzodiazepine, nicotine, cocaine or ethanol.
3. Use as claimed in Claim 1, in which said substance is a pharmaceutical.
4. Use as claimed in Claim 3, in which said agonist is to be administered before said pharmaceutical is first administered.
5. Use as claimed in Claim 3 in which said agonist is to be administered after said pharmaceutical has been administered.
6. Use as claimed in Claim 5, in which said agonist is to be administered after said pharmaceutical has been withdrawn.
7. Use as claimed in Claim 3, in which said agonist is to be co-administered with said pharmaceutical.
8. Use as claimed in Claim 3, in which said pharmaceutical is a benzodiazepine.
9. Use as claimed in Claim 8, in which said benzodiazepine is diazepam.
10. Use as claimed in Claim 2, in which said substance is nicotine.
11. Use as claimed in Claim 10, in which said mammal is ceasing smoking.
12. Use as claimed in Claim 1, in which said agonist acts selectively.
13. Use of an agonist which acts at negatively coupled cAMP-linked metabotropic glutamate receptors for the manufacture of metabotropic for the treatment of drug tolerance, withdrawal or cessation.
14. Use as claimed in Claim 13, in which said drug is an opiate, benzodiazepine, nicotine, cocaine or ethanol.
15. Use as claimed in Claim 14, in which the drug is a benzodiazepine.
16. Use as claimed in claim 15, in which the benzodiazepine is diazepam.
17. Use as claimed in Claim 13, in which said agonist acts selectively.
18. Use of an agonist which acts at negatively coupled cAMP-linked metabotropic glutamate receptors for the manufacture of a medicament for the treatment of smoking cessation.
19. Use as claimed in Claim 18, in which said agonist acts selectively.
20. Use of an agonist which acts at negatively coupled cAMP-linked metabotropic glutamate receptors for protecting a warm-blooded mammal from dependence on a dependence producing substance.
21. A pharmaceutical composition which comprises an agonist that acts at negatively coupled cAMP-linked metabotropic glutamate receptors, for use in protecting a warm-blooded mammal from dependence on a dependence-producing substance.
22. A pharmaceutical composition, which comprises an agonist that acts at negatively coupled cAMP-linked metabotropic glutamate receptors and a dependence-producing pharmaceutical.
Applications Claiming Priority (6)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US28995794A | 1994-08-12 | 1994-08-12 | |
| US08/289,957 | 1994-08-12 | ||
| US33734994A | 1994-11-10 | 1994-11-10 | |
| US08/337,349 | 1994-11-10 | ||
| US08/496,642 | 1995-06-29 | ||
| US08/496,642 US5661184A (en) | 1994-08-12 | 1995-06-29 | Psychiatric agents |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2195779A1 true CA2195779A1 (en) | 1996-02-22 |
Family
ID=27403948
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002195779A Abandoned CA2195779A1 (en) | 1994-08-12 | 1995-08-14 | Composition and method for protection against drug dependency |
Country Status (5)
| Country | Link |
|---|---|
| EP (1) | EP0776200A4 (en) |
| JP (1) | JP3618104B2 (en) |
| AU (1) | AU3325095A (en) |
| CA (1) | CA2195779A1 (en) |
| WO (1) | WO1996004900A1 (en) |
Families Citing this family (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1997018199A1 (en) * | 1995-11-16 | 1997-05-22 | Eli Lilly And Company | Excitatory amino acid derivatives |
| US5688826A (en) * | 1995-11-16 | 1997-11-18 | Eli Lilly And Company | Excitatory amino acid derivatives |
| EA000894B1 (en) * | 1995-11-16 | 2000-06-26 | Эли Лилли Энд Компани | substituted bicyclohexanecarboxylic acids and derivatives thereof as excitatory amino acid receptor antagonists, method for producing and use thereof |
| US5912248A (en) * | 1995-11-16 | 1999-06-15 | Eli Lilly And Company | Excitatory amino acid receptor antagonists |
| GB9605429D0 (en) * | 1995-11-16 | 1996-05-15 | Lilly Co Eli | Excitatory amino acid receptor antagonists |
| ZA969485B (en) * | 1995-11-16 | 1998-05-12 | Lilly Co Eli | Excitatory amino acid receptor antagonists. |
| ZA983930B (en) * | 1997-05-14 | 1999-11-08 | Lilly Co Eli | Excitatory amino acid receptor modulators. |
| JP4192331B2 (en) | 1999-04-16 | 2008-12-10 | 住友化学株式会社 | Process for producing optically active 2-oxobicyclo [3.1.0] hexane-6-carboxylic acid derivative |
| EP3661500A1 (en) | 2017-07-31 | 2020-06-10 | Novartis AG | Use of mavoglurant in the reduction of cocaine use or in preventing relapse into cocaine use |
-
1995
- 1995-08-14 WO PCT/US1995/010317 patent/WO1996004900A1/en not_active Application Discontinuation
- 1995-08-14 AU AU33250/95A patent/AU3325095A/en not_active Abandoned
- 1995-08-14 JP JP50756396A patent/JP3618104B2/en not_active Expired - Fee Related
- 1995-08-14 CA CA002195779A patent/CA2195779A1/en not_active Abandoned
- 1995-08-14 EP EP95929519A patent/EP0776200A4/en not_active Withdrawn
Also Published As
| Publication number | Publication date |
|---|---|
| EP0776200A4 (en) | 2001-01-17 |
| JP3618104B2 (en) | 2005-02-09 |
| EP0776200A1 (en) | 1997-06-04 |
| JPH10504038A (en) | 1998-04-14 |
| AU3325095A (en) | 1996-03-07 |
| WO1996004900A1 (en) | 1996-02-22 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US5661184A (en) | Psychiatric agents | |
| AP758A (en) | Synthetic excitatory amino acids. | |
| RU2001614C1 (en) | R(+)-n-propargyl-1-aminoindan and its pharmaceutically acceptable acid-additive salts, and pharmaceutical composition showing inhibitory activity relative to b-form of enzyme monoamineoxidase | |
| EP0776201B1 (en) | Treating anxiety | |
| CA2195779A1 (en) | Composition and method for protection against drug dependency | |
| JPH11509846A (en) | Heterocyclic compounds, their preparation and use | |
| CA2285192A1 (en) | Pharmacological agents | |
| US5521174A (en) | N-acyl-2,3-benzodiazepine derivatives and a method of treating spasms of the skeletal musculature therewith | |
| EP1512679B1 (en) | Novel adamantane derivatives with neuroprotective, antidepressant and anti-ischaemic activities, and process for preparing them | |
| EP1194400A1 (en) | 2-aminoindane analogs | |
| Tencé et al. | Synergistic effects of acetylcholine and glutamate on the release of arachidonic acid from cultured striatal neurons | |
| WO2001032176A1 (en) | Beta-carboline derivatives for use as analgesics | |
| EP1194399A2 (en) | Novel aminoindanes | |
| JP3023987B2 (en) | (S) -α-phenyl-2-pyridineethanamine (S) -malate and its use as a medicament | |
| EP0928792B1 (en) | Bicyclic(3.1.0)hexanes and related compounds | |
| IL133493A (en) | 2-spiro-5'-hydantoinbicyclo [3.1.0] hexane-6-carboxylic acid derivatives | |
| WO2013184732A1 (en) | Crystalline (2s)-(4e)-n-methyl-5-[3-(5-isopropoxypyridin)yl]4-penten-2-amine 1,5-naphthalenedisulfonate | |
| WO2004024709A2 (en) | Xanthenyl cubane analogs with activity at the metabotropic glutamate receptors | |
| CA2407777A1 (en) | Novel spiro¬2.4|heptane amino carboxy compounds and derivatives thereof | |
| MXPA99009140A (en) | Pharmacological agents |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| EEER | Examination request | ||
| FZDE | Dead |