WO2008004067A2 - Alpha-2-delta-1 selective compounds for disorders of the nervous system - Google Patents

Alpha-2-delta-1 selective compounds for disorders of the nervous system Download PDF

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Publication number
WO2008004067A2
WO2008004067A2 PCT/IB2007/001789 IB2007001789W WO2008004067A2 WO 2008004067 A2 WO2008004067 A2 WO 2008004067A2 IB 2007001789 W IB2007001789 W IB 2007001789W WO 2008004067 A2 WO2008004067 A2 WO 2008004067A2
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Prior art keywords
disorder
delta
compound
alpha
selective
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PCT/IB2007/001789
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French (fr)
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WO2008004067A3 (en
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Zheng Li
Jacob Bradley Schwarz
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Pfizer Products Inc.
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Publication of WO2008004067A3 publication Critical patent/WO2008004067A3/en

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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
    • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P15/00—Drugs for genital or sexual disorders; Contraceptives
    • A61P15/12—Drugs for genital or sexual disorders; Contraceptives for climacteric disorders
    • 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
    • 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
    • A61P25/02—Drugs for disorders of the nervous system for peripheral neuropathies
    • 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
    • A61P25/04—Centrally acting analgesics, e.g. opioids
    • 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
    • A61P25/08—Antiepileptics; Anticonvulsants
    • 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
    • A61P25/08—Antiepileptics; Anticonvulsants
    • A61P25/10—Antiepileptics; Anticonvulsants for petit-mal
    • 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
    • A61P25/14—Drugs for disorders of the nervous system for treating abnormal movements, e.g. chorea, dyskinesia
    • 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
    • A61P25/20—Hypnotics; Sedatives
    • 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
    • A61P25/22—Anxiolytics

Definitions

  • the invention is directed to methods of treating disorders or conditions by administering a selective ligand preferring alpha-2-de!ta-1 subunit of calcium channel membrane proteins.
  • liga ⁇ ds are useful for treating CNS and pain-related disorders and conditions and may avoid or reduce adverse effects associated with administration of nonselective alpha-2-delta liga ⁇ ds.
  • Voltage-gated calcium channels are formed by combinations of the pore-forming al ⁇ ha-1 ( ⁇ ,) subunit, and auxiliary alpha-2-delta, beta, and gamma (c-:_ ⁇ , ⁇ and ⁇ , respectively) proteins (Cattera ⁇ W.A. (2000) Annul. Rev. Cell Dev. Biol. 16: 521-555).
  • the oc 2 ⁇ protein is known to regulate both the calcium channel density and voltage-dependent kinetics of these calcium channels (Felix R. ef a/. (1997) J, Neur ⁇ sci. 17: 6884-6891; Klugbauer N, etal. (1999) J. Neurosci. 19: 684-691; Hobom M.
  • the OG 2 S protein is encoded by a single gene and post-translationally cleaved to ⁇ 2 and ⁇ subunits.
  • the ⁇ 2 subunit is a highly glycosylated extracellular protein and is associated with the membrane anchor protein ⁇ by disulfide linkage (Wang M, ef a/. (1999) Biochern. J. 342:313-320; Marais E. et a!. (2001) MoI. Pharmacol.
  • WO 00/20450 which refers to c-z ⁇ -1, ⁇ z ⁇ -2, ⁇ -3, and ⁇ 2 ⁇ -4 as ⁇ a ⁇ -A, O -2 S-B 1 ct2 ⁇ -C, and «28- D, respectively.
  • GenbanK accession numbers for ⁇ 2 ⁇ -1 subunits include: M76559 (human); U73483, U734S4, U73485, U73486, and U73487 (mouse); M86S21 (rat); and AF077 ⁇ 65 (pig). See also Brown and N. Gee. (1998) J. Biol. Chem, 273:25458-25465.
  • the accession numbers for ⁇ 2 ⁇ -2 subunits include: AJ251368, AJ251367.1 , Abo11130.1 , NM-
  • AF040709 AF042792, AF042793 (human); AF247139, NM-02063.2. AF247141, AB093246.1, AK0446D3.1 (mouse); and AF486277.1 and NM-175592.2 (rat).
  • Non-selective ⁇ S ligands bind with high affinity to two of the subtypes of calcium channel ⁇ 2 ⁇ subtypes.
  • Gabapenti ⁇ , 1-ami ⁇ omethyl-oyclohexyl-acetic acid (Neurontin ® ) is recognized as an anti-epileptic drug which binds with high affinity to ⁇ H ⁇ -1 and ⁇ 2 ⁇ -2.
  • Pregabslin, S-(+) 3-aminomethyl-5-methyl-hexanoic acid (Lyrica 10 ) also binds with high affinity to the C-2S-I and ⁇ 2 ⁇ -2 subtypes.
  • Gabape ⁇ ti ⁇ and Pregabalin are co ⁇ sldered to be non-selective ⁇ 2 ⁇ ligands (Gong H.C. ef a/. (2001) J, Membr. Biol. 184:35- 43). •
  • This invention relates to methods of treating disorders or conditions in a mammal, including a human, comprising administering to said mammal a therapeutically effective amount of an aIpha-2-delta-1 ( ⁇ & ⁇ -l) selective ligand, or a pharmaceutically acceptable salt thereof, wherein said disorder or condition is selected from pain, fibromyalgia, epilepsy, ] 0 restless legs syndrome, hot flashes, mood disorders and sleep disorders.
  • an oa ⁇ -i selective ligand avoids or reduces one or more adverse effects associated with the administration of an o ⁇ non-selective ligand, including but not limited to sedation, dizziness, ataxia, and asthenia.
  • the avoidance or reduction of adverse effects following the administration of an ⁇ 2 ⁇ -1 selective ligand is 15 attributed to the high binding affinity of the c- a ⁇ -l selective ligand to the calcium channel subu ⁇ it cfe ⁇ -1 subtype.
  • the present invention is directed to methods of treating a disorders or conditions in a mammal, including a human, comprising administering to said mammal a therapeutically effective amount of an alpha-2-delta-1 (0 5 .6-1) selective ligand, or a pharmaceutically acceptable salt thereof.
  • ligands are useful far treating central nervous system (CNS) and pain-related disorders and conditions and may avoid or reduce adverse effects
  • the CNS and pain-related disorders include, but are not limited to pain, fibromyalgia, epilepsy, restless legs syndrome, hot flashes, mood disorders, and steep disorders.
  • the adverse effects include but are not limited to sedation, dizziness, ataxia, and asthenia.
  • the degree of binding to the O -2 S subunit can be determined using the radioligand
  • ⁇ 2 ⁇ -l selective ligand means an O 3 S-I selective ligand, or pharmaceutically acceptable salt thereof, wherein said ligand or salt is at least 30-fold more effective in displacing bound [ 3 H]-gabapentin from the ⁇ j ⁇ -1 subunit than the ⁇ s ⁇ -2 subunit; as determined by comparing the binding inhibition
  • Kt of the ligand for displacing [ 3 H]-gabapentin bound to the ⁇ j ⁇ -2 subu ⁇ it Kt of the ligand for displacing [ 3 H]-gabapentin bound to the ⁇ j ⁇ -2 subu ⁇ it.
  • selective is in reference to the ration of Ki values of a ligand for displacing [ a H]-gabapentin bound to the ⁇ -1 subunit in comparison to the Ki value of a compound for displacing [ 3 H]-gabapenti ⁇ bound to the ⁇ 2 ⁇ -2 subunit.
  • Cx 3 S-I selective ligands can be identified among test compounds by comparing the K-, of the test Iigand for displacing [ 3 H]-gabapentin bound to the ⁇ 2 ⁇ -1 subunit, to the & of the compound for displacing [ s H]-gabapenti ⁇ bound to the a ⁇ b-i subunit.
  • Alpha-2-delta-1 ligands may include those compounds that are generally or specifically disclosed in the following references: US4024175, EP0641330, including 3-methylgabape ⁇ tin; US5563175, WO97/33858, WO97/33859, WQ/99/31057,
  • WO99/31074 WO97/291Q1, WO02/0SS839, including [(1R,5R,6S)-6- (ami ⁇ omethy!)bicyclo[3,2.0]hept-6-yl]acetio acid
  • WO99/31075 including 3- ⁇ 1-aminomethyl- cyclohexylmethyl)-4H-[1 ,2,4]oxadiazol-5-one and C-[1-(1H-tetrazo!-5-y)methyl)-cycloheptyl]- methylamine
  • WO99/21624 including (3S,4S)-(1-aminomethyi-3,4-dimethyl-oyclopenty[)- acetic acid
  • WO01/90Q52 WO01/28978, including (1 ⁇ ,3 ⁇ ,5 ⁇ ) ⁇ 3-amino-methyl- bicyc
  • WOS8/17627 WO
  • Alpha-2-delta ligands include those depicted by the following formula (I):
  • X is a carboxylic acid or carboxylic acid bioisostere; n is 0, 1 or 2; and R 1 , R 1a , R 2 , R 2a , R 3 , R 3a , R ⁇ and R 4a are independently selected from H and C 1 -C 6 aikyi, or R 1 and R 2 , or R 2 and R 3 , are taken together to form a C 3 -C 7 cycloalkyl ring, which is optionally substituted with one or two substituents selected from C 1 -Ce aikyl, or a pharmaceutically acceptable salt or solvate thereof.
  • X is preferably a carboxylic acid.
  • R 1 , R 1a , R 2a , R 3* , R 4 and R 43 are H and R 2 and R 3 are independently selected from H and methyl, or R 1a , R 2a , R 3a and R 4a are H and R 1 and R 2 or R 2 and R 3 are taken together to form a C 4 -C 5 cycloalkyl ring, or, when ⁇ is 0, R 1 , R 1a , R 2B , R 3a , R 4 and R" 3 are H and R 2 and R 3 form a cydopentyl ring, or, when n is 1, R 1 , R 1a , R 2a , R 33 , R 4 and R* 1 are H and R 2 and R 3 are both methyl or R 1 , R 1a , R 2 *, R 3a , R 4 and R 4a are H and R 2 and R 3 form a cyclobutyl ring, or, when n is 2, R 1 , R 1a , R
  • Alpha2delta ligands also include those depicted by the following formula (II):
  • n is 0 or 1
  • R n is hydrogen or C 1 -C 6 alkyl
  • R 2 is hydrogen or CrC 6 aikyl
  • R 3 is hydrogen gr C 1 -Ce aikyl
  • R 4 is hydrogen or C 1 -C 5 alkyl
  • R s is hydrogen or CrC 6 alkyl
  • R 2 is hydrogen or C 1 -Ca alkyl, or a pharmaceutically acceptable salt, ester or solvate thereof.
  • 0- 2 6 ligands are Compounds of formula (II) wherein R 1 is Ci-C 8 alkyl, R 2 is methyl, R 3 - R 8 are hydrogen and ⁇ is 0 or 1,
  • O 2 S ligands are Compounds of formula (H) wherein R 1 is methyl, ethyl, n-propyl or n-butyl, R 2 is methyl, R 9 -
  • R 6 are hydrogen and n is 0 or 1.
  • R 3 — R 8 are hydrogen and n is 0,
  • R 1 is suitably ethyl, n-propyl or ⁇ -butyl.
  • R 2 is methyl
  • R 3 - R 6 are hydrogen and n is 1
  • R 1 is suitably methyl or n-propyl.
  • Compounds of formula (II) are suitably in the 3S, 5R configuration.
  • Al ⁇ ha-2-deIta ligands also include: f(1R,5R,6S)-6-(ami ⁇ omethyl)bi ⁇ ycle-[3.2.0]hept-6- yl]acetic acid, 3-(1-aminomethyl-cyclohexylmethyl)-4H-[1,2,4]oxadlazol-5-one, C-[1-(1H- tetrazoI-5-ylrnethyl)-cydoheptyl]-methylami ⁇ e, (3S,4S)-(1-amJ ⁇ ornethyl-3.4-dimethyl- cyclopentyl)-acetig acid, (1 ⁇ ,3 ⁇ I 5 ⁇ )(3-ami ⁇ o-methy!-bicyclo[3.2 ⁇ O]h ⁇ pt-3-yl)-acetic acid, (3S,5R)-3-aminomethyl ' 5-methyl-octa ⁇ o ⁇ e acid, (3S,5R)-3-amino
  • c f cS nonselective ligands include gabapenti ⁇ (3S, 5R)-3-ami ⁇ methyI-6-cydopropyl-5-methyl- hexanoic acid, and pregabalin, S-(+) e ⁇ antiomer of 3-aminomethyl-S-methyl-hexanoic acid (Lyrica ® ).
  • an Ot 3 S-I selective ligand, (3S,4R,5R)-3- Arnino-4,5-dimethyloc!tanoic acid hereinafter "Compound A" i$ described.
  • Selectivity is determined by calculating the binding affinity Ki value from the amount °f [ 3 H]-gabapentin bound to calcium channel a ⁇ subu ⁇ it membrane proteins prepared from both recombinant porcine (X 2 S-I cells and recombinant human ⁇ 2 ⁇ -2 cells.
  • the K 1 values for displacement of binding of [ 3 H]-gabapentin to recombinant porcine ⁇ 2 ⁇ -l and recombinant human a. ⁇ -2 cell membranes correspond to the binding affinity of the Qt 2 S ligands.
  • values of the two recombinant cell membrane proteins corresponds to binding selectivity toward ct a S-1.
  • Selectivity of an oc 2 ⁇ -1 selective ligand to the O 8 M subtype is at least 30-, 40-, 50-, 100-, 500-, or 1000-fold more than for binding to the ⁇ a ⁇ -2 subunit
  • the mean K 1 values for Compound A are 35 nM and 1670 nM for recombinant porcine ⁇ j ⁇ -1 and recombinant human ⁇ 2 ⁇ -2 cell membranes, respectively, accounting for a 48-fold selectivity for the Ct 2 S-I subunit
  • ⁇ a ⁇ -1 selective ligand or a pharmaceutically acceptable salt thereof, in an amount that is therapeutically effective to treat one or more of the disorders or conditions described herein.
  • therapeutically effective amount will generally be from about 1 to about 300 mg/kg of subject body weight
  • Typical doses will be from about 10 to about 5000 mg/day for an adult subject of normal weight
  • regulatory agencies such as, for example, the Food and Drug Administration ("FDA") in the US, may require a particular therapeutically effective amount
  • FDA Food and Drug Administration
  • a number of factors will generally be considered by the medical practitioner or veterinarian in view of the experience of the medical practitioner or veterinarian, published clinical studies, the subject's age, sex, weight and genera!
  • the administered dose rnay fall within the ranges or concentrations recited above, or may vary outside, i.e., either below or above, those ranges depending upon the requirements of the individual subject, the severity of the condition being treated, and the particular therapeutic formulation being employed, Determination of a proper dose for a particular situation is within the skill of the medical or veterinary arts, Generally, treatment may be initiated using smaller dosages of the Ct 2 S-I iigand that are less than optimum for a particular subject. Thereafter, the dosage can be increased by small increments until the optimum effect under the circumstance is reached. For convenience, the total daily dosage may be divided and administered in portions during the day, if desired.
  • a preferred embodiment relates to a method of treating a disorder or condition selected from the group consisting of pain such as acute pain, chronic pain, pain resulting from soft tissue and peripheral damage such as acute trauma; complex regional pain syndrome also referred to as reflex sympathetic dystrophy, postherpetic neuralgia, occipital neuralgia, trigeminal neuralgia, segmental or intercostal neuralgia and other neuralgias; musculoskeletal pain such as pain associated with strains, sprains and trauma such as broken bones; central nervous system pain such as pain associated with spinal pain, spinal pain associated wfth cord or brain stem damage; lower back pain, sciatica, dental pain, myofascial pain syndromes, episiotomy pain, gout pain, and pains resulting from burns; deep and visceral pain such as heart pain, musde pain, eye pain, inflammatory pain, orofascial pain, for example odontalgia; abdominal pain, and gynecological pain, for
  • Another preferred embodiment relates to a method of treating a disorder or condition selected from the group consisting c-f mood disorders, such as depression, or more particularly, depressive disorders, for example, single episodic or recurrent major depressive disorder, severe unipolar recurrent major depressive episodes, and melancholic depression; seasonal affective disorder, conduct disorder and disruptive behavior disorder, obsessive compulsive disorder, stress related somatic disorders and anxiety disorders, for example, generalized anxiety disorder, social anxiety disorder; stress disorders including post traumatic stress disorder and acute stress disorder in a mammal, comprising administering to a mammal in need of such treatment a therapeutically effective amount of an O 2 S-I ligand, or a pharmaceutically acceptable salt thereof.
  • a disorder or condition selected from the group consisting c-f mood disorders, such as depression, or more particularly, depressive disorders, for example, single episodic or recurrent major depressive disorder, severe unipolar recurrent major depressive episodes, and melancholic depression; seasonal affective disorder, conduct disorder and disruptive
  • insomnia ⁇ e.g., primary insomnia including psychophysiological and idiopathic insomnia, secondary insomnia including insomnia secondary to restless legs syndrome, insomnia secondary to anxiety, insomnia secondary to fibromyalgia, insomnia secondary to pain and neuropathic pain, and transient insomnia), sleep deprivation, REM sleep disorders, sleep apnea, hypersomnia, parasomnras, sleep-wake cycle disorders, jet lag, narcolepsy, sleep disorders associated with shiftwork or irregular work schedules, deficient sleep quality due to a decrease in slow wave sleep caused by medications or other sources, and other sleep disorders in a mammal in need of such treatment a therapeutically effective amount of an Qt 2 S-I ligand, or a pharmaceutically acceptable salt thereof.
  • insomnia ⁇ e.g., primary insomnia including psychophysiological and idiopathic insomnia, secondary insomnia including insomnia secondary to restless legs syndrome, insomnia secondary to anxiety, insomnia secondary to fibromyalgia, insomnia secondary to pain and neuropathic pain, and transient insomnia
  • Fibromyalgia is a chronic syndrome characterized mainly by widespread pain, un-refreshing sleep, disturbed mood, and fatigue. Syndromes commonly associated with FWl include irritable bowel syndrome, and migraine headaches, among others. Success of treating FWl with a single pharmacological agent has been characterized as modest and results of clinical trials have been characterized as disappointing. It is believed that based on current understanding of the mechanisms and pathways involved in FM, multiple agents will be required, aimed at the major symptoms of pain, disturbed sleep, mood disturbances, and fatigue.
  • Fibromyalgia patients are often sensitive to side effects of medications, a characteristic perhaps related to the pathophysiology of this disorder (Barkhuizen A, Rational and Targeted pharmacologic treatment of fibromyalgia Rheum Dis Clin N Am 2002; 28: 261-290; Leventhal LJ. Management of fibromyalgia, Ann intern Med 1999;131:850-S).
  • FM is a complex disorder with multiple facets, this complexity can be well assessed (Yunus MB, A comprehensive medical evaluation of patients with fibromyalgia syndrome, Rheum Dis N Am 2002; 28:201-217). The diagnosis of FM is usually based on the
  • Another preferred embodiment relates to a method of treating a disorder or condition selected from the group consisting of epilepsy.
  • Epilepsy is an episodic disturbance of consciousness during which generalized convulsions may occur. The condition is of unknown etiology, often hereditary, and is manifested by symptoms of a peculiar sensation, smell or feeling, called an "aura" proceeding the loss of consciousness and often convulsions.
  • epileptic seizures in a mammal, including a human, are classified on the basis of the clinical manifestation ⁇ of the onset and the EKG pattern. Sei ⁇ ure types are often used to classify the particular type or types of epilepsy which include gran mal (generalized t ⁇ nic- chronic), absence (petit mal), cortical focal, temporal lobe (psychomotor), and infantile that often occur in adolescents.
  • Restless legs syndrome is a common, potentially disabling condition that affects about 10% to 15% of the general population and yet is often unrecognized and misdiagnosed. It is mainly diagnosed clinically and only rarely requires polysomnography. The condition is usually primary and treatable. Restless legs syndrome is a chronic condition. Symptoms may worsen with age, and the most disabling feature is sleep onset insomnia. Restless legs syndrome is a sensory-motor
  • Hot flashes occur in both male and females mammals, including humans.
  • Females having a low level of estrogen are prone to suffer from hot flashes.
  • This deficiency can be due to radiation therapy, which can prematurely induce menopause, or pan be caused by specific medications such as anti-estrogen treatment or certain drugs (e.g. Tamoxifen (Nolvadex)).
  • hot flashes may be secondary to menopause or postmenopause, medical treatment, and cancer. In men, androgen deprivation can be a cause of hot flashes.
  • hormone imbalance can be drug-induced
  • Another preferred embodiment relates to a method of treating a disorder or condition selected from the invention methods, comprising administration of a therapeutically effective amount of an ⁇ 2 ⁇ -1 ligand, or a pharmaceutically acceptable salt thereof, to a mammal, including a human, wherein at least one adverse effect is avoided or reduced.
  • Another embodiment relates to a method of neuroprotection for brain damage caused by stroke, cardiac arrest, Alzheimer's disease, and other related conditions.
  • Another embodiment relates to any of the above methods wherein the « 5 ,6-1 ligand, or a pharmaceutically acceptable salt thereof, is administered to a mammal, including a human, for the treatment of any two or more comorbid disorders and conditions referred to in any of the above methods.
  • Another embodiment relates to any of the above methods for treating any of the invention methods, comprising administration of an ⁇ j ⁇ -1 selective ligand, wherein the said ligand is (3S,4R,5R)-3-Amino-4,5-dimethylocta ⁇ oic acid or a pharmaceutically acceptable salt thereof, to a mammal, including a human.
  • Another embodiment relates to adverse effects being avoided or reduced in a mammal, including a human, comprising the administration of an ⁇ .a ⁇ -1 selective ligand, or pharmaceutically acceptable salt thereof.
  • abnormal vision includes, but is not limited to: dizziness, asthenia, ataxia, sedation, somnolence, fatigue, nystagmus, weight gain, ernesis, peripheral edema, dyspepsia, tremor, nervousness, amnesia, depression, twitching, myalgia, rhinitis, diplopia, amblyopia, malaise, hypertension, flatulence, purpura most often described as bruises resulting from physical trauma, arthralgia, vertigo, hyperkinesia, decreased or absent reflexes, increased reflexes, hostility, and abnormal vision.
  • avoid means that the adverse effect does not occur in the mammal following administration of an o.j ⁇ -1 selective liga ⁇ d
  • the term “reduce”, “reduces”, or “reduced” as used herein means that the frequency or incidence of one or more adverse effects has been decreased by at least 10 percent.
  • Another embodiment relates to the administration of an Os 2 S-I selective ligand in an amount of about 10 to 5000 mg per day.
  • Another embodiment relates to the administration of an Cx 2 S-I selective ligand that is in a liquid or solid dosage form.
  • Administration of a dosage form comprising an ⁇ a ⁇ -1 selective ligand. or a pharmaceutically acceptable salt thereof may be administered orally, parenteral ⁇ , subcutaneously, intravenously, intramuscularly, intraperitoneally, intracerebroventricularly, by intranasal instillation, by implantation, by intracavitary or intravesical instillation, i ⁇ traocularly, intraarterially, i ⁇ tralesionaliy, intracerebroventricular, or by application to dermal, sublingual, or mucous membranes.
  • compositions comprising an O 2 S-I selective ligand and a pharmaceutically acceptable carrier.
  • pharmaceutically acceptable refers to those properties and/or substances which are acceptable to the mammal, including a human, from a pharmacological and toxicological perspective regarding composition, formulation, stability, safety, and bioavailability,
  • compositions of an O 2 S-I ligand, or a pharmaceutically acceptable salt thereof are produced by formulating the active compound in dosage unit form with a pharmaceutical carrier.
  • dosage unit forms are tablets, capsules, pills, powders, aqueous and nonaqueous oral solutions and suspensions, transdermals, and parenteral solutions packaged in containers containing either one or some larger number of dosage units and capable of being subdivided into individual doses.
  • suitable pharmaceutical carriers including pharmaceutical diluents
  • suitable pharmaceutical carriers are gelatin capsules; sugars such as lactose and sucrose; starches such as corn starch and potato starch; cellulose derivatives such as sodium carboxymethyl cellulose, ethyl cellulose, methyl cellulose, and cellulose acetate phthaiate; gelatin; talc; stearic acid; magnesium stearate; vegetable oils such as peanut oil, cottonseed oil, sesame oil, olive oil, corn oil, and oil of theobroma; propylene glycol, glycerin; sorbitol; polyethylene glycol; water; agar; alginic acid; isotonic saline, and phosphate buffer solutions; as well as other compatible substances normally used in pharmaceutical formulations,
  • compositions to be employed in the invention can also contain other components such as coloring agents, flavoring agents, and/or preservatives. These materials, if present, are usually used in relatively small amounts.
  • the compositions can, if desired, also contain other therapeutic agents commonly employed to treat the disorder or condition being treated.
  • the percentage of the active ingredients in the foregoing compositions can be varied within wide limits, but for practical purposes it is preferably present in a concentration of at least 10% in a solid composition and at least 2% in a primary liquid composition. The most ⁇ atisfactory compositions are those in which a much higher proportion of the active ingredient is present, for example, up to about 95%.
  • the OC 2 S-I ligand, or a pharmaceutically acceptable salt thereof may be administered in any form.
  • administration is in unit dosage form
  • a unit dosage form of the ⁇ 2 ⁇ -l ⁇ ga ⁇ d, or a pharmaceutically acceptable salt thereof, to be used in this invention may also comprise other compounds useful in the therapy of the disorder or condition for which the ⁇ 2 ⁇ - 1 ligand is being administered or a disorder or condition that is secondary to the disorder or treatment for which the ⁇ a ⁇ -1 iiga ⁇ d is being administered.
  • Some of the compounds utilized in a method of the present invention are capable of further forming pharmaceutically acceptable salts, including, but not limited to, acid addition and/or base salts.
  • the acid addition salts are formed from basic compounds, whereas the base addition salts are formed from acidic compounds.
  • AIi of these forms are within the scope of the compounds useful in the method of the present invention.
  • Pharmaceutically acceptable acid addition salts of the basic compounds useful in the method of the present invention include nontoxic salts derived from inorganic acids such as hydrochloric, nitric, phosphoric, sulfuric, hydr ⁇ brom ⁇ c, hydroiodic, hydrofluoric, phosphorous, and the like, as well nontoxic salts derived from organic acids, such as aliphatic mono- and dicarboxylic acids, phenyl-substituted alkartoic acids, hydroxy alkanoio acids, alkanedioic acids, aromatic acids, aliphatic and aromatic sulfonic acids, etc.
  • inorganic acids such as hydrochloric, nitric, phosphoric, sulfuric, hydr ⁇ brom ⁇ c, hydroiodic, hydrofluoric, phosphorous, and the like
  • organic acids such as aliphatic mono- and dicarboxylic acids, phenyl-substituted alkartoic acids, hydroxy alkanoio acids, alkanedi
  • Such salts thus include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, m ⁇ ohydrogenphosphate, dihydrogenphosphate, r ⁇ etaphosphste, pyrophosphate, chloride, bromide, iodide, acetate, trifluoroacetate, propionate, capryiate, isobutyrate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, mandelate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, phthalate, benzenesulf ⁇ nate, t ⁇ lue ⁇ esulfonate, phenylacetate, citrate, lactate, malate, tartrate, methanesulfo ⁇ ate, and the like.
  • salts of amino acids such as arginate and the like and gluconate, galacturonate (see, for example, Berge S. Wl, et a!., "Pharmaceutical Salts,” J, otPharma. ScL, 1977;66:1).
  • An acid addition salt of a basic compound useful in the method of the present invention is prepared by contacting the free base form of the compound with a sufficient amount of a desired acid to produce a nontoxic salt in the conventional manner.
  • the free base form of the compound may be regenerated by contacting the acid addition salt so formed with a base, and isolating the free base form of the compound in the conventional manner,
  • the free base forms of compounds prepared according to a process of the present invention differ from their respective acid addition salt forms somewhat in certain physical properties such as solubility, crystal structure, hygroscopicity, and the like, but otherwise free base forms of the compounds and their respective acid addition salt forms are equivalent for purposes of the present invention.
  • a pharmaceutically acceptable base addition salt of an acidic compound useful in the method of the present invention may be prepared by contacting the free acid form of the compound with a nontoxic metal cation such as an alkali or alkaline earth metal cation, or an amine, especially an organic amine.
  • a nontoxic metal cation such as an alkali or alkaline earth metal cation, or an amine, especially an organic amine.
  • suitable metal cations include sodium cation
  • Suitable amines are N.N'-dibenzylethyle ⁇ ediamine, chloroprocai ⁇ e, choline, diethanolamine, dicyclohexylamine, ethylenediami ⁇ e, N-methylglucamine, and procaine (see, for example, Berge, supra., 1977).
  • a base addition salt of an acidic compound useful in the method of the present invention may be prepared by contacting the free acid form of the compound with a sufficient amount of a desired base to produce the salt in the conventional manner.
  • the free acid form of the compound may be regenerated by contacting the salt form so formed with an acid, and isolating the free acid of the compound in the conventional manner.
  • the free acid forms of the compounds useful in the method of the present invention differ from their respective salt forms somewhat in certain physical properties such as solubility, crystal structure, hygrosc ⁇ p ⁇ city, and the like, but otherwise the saits are equivalent to their respective free acid for purposes of the present invention.
  • Certain of the compounds useful in the method of the present invention can exist in unsolvated forms as well as solvated forms, including hydrated forms. In general, the solvated forms, including hydrated forms, are equivalent to unsolvated forms and are intended to be encompassed within the scope of the present invention. Certain of the compounds useful in the method of the present invention possess one or more stereoce ⁇ ters, and each center may exist in the R or S configuration. A method of the present invention may utilize any diastereomeric, enantiomeric, or epimeric form of an ⁇ 2 ⁇ -1 ligand, or a pharmaceutically acceptable salt thereof, as well as mixtures thereof,
  • certain compounds useful in the method of the present invention may exist as geometric isomers such as the
  • E and
  • Z isomers of alke ⁇ yl groups.
  • a method of the present invention may utilize any cis, trans, syn, anti,
  • E and
  • Z isomer of an ⁇ j ⁇ -1 ligand, or a pharmaceutically acceptable salt thereof, as well as mixtures thereof.
  • Certain compounds useful in the method of the present invention can exist as two or more tautomeric forms. Tautomeric forms of the compounds may interchange, for example, via enol ⁇ zatio ⁇ /de-e ⁇ olization and the like.
  • a method of the present invention may utilize any tautomeric form of an ⁇ 2 ⁇ -1 ligand, or a pharmaceutically acceptable salt thereof, as well as mixtures thereof.
  • pharmaceutically acceptable carriers can be either solid or liquid. Solid form preparations include powders, tablets, pills, capsules, cachets, suppositories, and dispersible granules.
  • a solid carrier can be one or more substances wnich may also act as diluents, flavoring agents, binders, preservatives, tablet disintegrating agents, or an encapsulating material.
  • the carrier is a finely divided solid which is in a mixture with the finely divided active component.
  • the active component is mixed with the carrier having the necessary binding properties in suitable proportions and compacted in the shape and size desired.
  • the powders and tablets preferably contain from five or ten to about seventy percent of the active compound, Suitable carriers are magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, tragaca ⁇ th, methyleeltulose, sodium carbo ⁇ ymethylcellulose, a low melting wax, cocoa butter, and the like.
  • preparation is Intended to include the formulation of the active compound with encapsulating material as a carrier providing a capsule in which the active component, with or without other carriers, is surrounded by a carrier, which is thus in association with it.
  • carrier providing a capsule in which the active component, with or without other carriers, is surrounded by a carrier, which is thus in association with it.
  • cachets and lozenges are included. Tablets, powders, capsules, pills, cachets, and lozenges can be used as solid dosage forms suitable for oral administration.
  • a low melting wax such as a mixture of fatty acid glycerides or cocoa butter
  • the active component is dispersed homogeneously therein, as by stirring.
  • the molten homogenous mixture is then poured into convenient sized molds, allowed to cool, and thereby to solidify.
  • Liquid form preparations include solutions, suspensions, and emulsions, for example, water or water propylene glycol solutions.
  • liquid preparations can be formulated in solution in aqueous polyethylene glycol solution.
  • Aqueous solutions suitable for oral use can be prepared by dissolving the active component in water and adding suitable colorants, flavors, stabilizing and thickening agents as desired.
  • Aqueous suspensions suitable for oral use can be made by dispersing the finely divided active component in water with viscous material, such as natural or synthetic gums, resins, methylcellulose, sodium carboxymethyleellulose, and other well-known suspending agents.
  • viscous material such as natural or synthetic gums, resins, methylcellulose, sodium carboxymethyleellulose, and other well-known suspending agents.
  • solid form preparations which are intended to be converted, shortly before use, to liquid form preparations for oral administration.
  • liquid forms include solutions, suspensions, and emulsions.
  • These preparations may contain, in addition to the active component, colorants, flavors, stabilizers, buffers, artificial and natural sweeteners, dispersants, thickeners, solubilizing agents, and the like.
  • the pharmaceutical preparation is preferably in unit dosage form. In such form the preparation is subdivided into unit doses containing appropriate quantities of the active -J 4-
  • the unit dosage form can be a packaged preparation, the package containing discrete quantities of preparation, such as packeted tablets, capsules, and powders In vials or ampoules. Also, the unit dosage form can be a capsule, tablet, cachet, or lozenge itself, or it can be the appropriate number of any of these in packaged form.
  • the quantity of active component in a unit dose preparation may be varied or adjusted from 0.1 mg to 1 g according to the particular application and the potency of the active component In medical use the drug may be administered three times daily as, for example, capsules of 100 or 300 mg.
  • the composition can, if desired, also contain other compatible therapeutic agents.
  • the compounds utilized in the pharmaceutical method of this invention are administered at the initial dosage of about 0.01 mg to about 100 mg/kg daily.
  • a daily dose range of about 0.01 mg to about 100 mg/kg is preferred.
  • the dosages may be varied depending upon the requirements of the patient, the severity of the condition being treated, and the compound being employed. Determination of the proper dosage for a particular situation is within the skill of the art. Generally, treatment is initiated with smaller dosages which are less than the optimum dose of the compound. Thereafter, the dosage is increased by small increments until the optimum effect under the circumstances is reached. For convenience, the total daily dosage may be divided and administered in portions during the day, if desired.
  • treating refers to reversing, alleviating, inhibiting the progress of, or preventing the disorder or condition to which such term applies, or preventing one or more symptoms of such condition or disorder.
  • treatment refers to the act of treating, as “treating” is defined immediately above.
  • terapéuticaally effective refers to the treatment of a mammal, including a human, with an amount of an ⁇ 2 ⁇ -1 ⁇ gand, or a pharmaceutically acceptable salt thereof, to treat one or more of the invention disorders or conditions as described herein.
  • lower alkyl means a straight or branched alkyl group or radical having from 1 to 6 carbon atoms, and includes methyl, ethyl, n-propyl, /-propyl, ⁇ -butyl, /-butyl, sec- butyl, ferf-butyl, ⁇ -pentyl, n-hexyl, and the like.
  • alky as used herein, unless otherwise indicated, includes saturated monovalent hydrocarbon radicals having straight, branched or cyclic moieties or combinations thereof.
  • alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, iso- sec- and tert-butyl, pentyl, hexyl, heptyl, 3-ethylbutyl, cyclopropyl, cyclobutyl, cyclopentyl, cycloh ⁇ xyl, cycloheptyl, norbornyl, and the like,
  • the cycloalkyl groups are saturated monovalent carbocyclic groups containing from 3 to 7 carbons and are selected from cyclopr ⁇ pyl, cyclobutyl, cyclope ⁇ tyl, cyclohexyl, and cycloheptyi, unless otherwise stated.
  • this invention relates to methods of treatment utilizing the O 2 S-I selective liga ⁇ d, ⁇ 3$,4R,5R)-3-Amino-4,5-dirnetr ⁇ yloctanoic acid (hereinafter
  • Compound A or a pharmaceutically acceptable salt thereof; which compound can be prepared by:
  • the 20 L reactor was fitted with a distillation head and the organic layer was distilled to remove in succession: 13.5 L of EtOAc, after which 5 L of heptane was added to the reactor; 5 L of EtOAc/heptane, after which 5 L of heptane was added to the reactor; and 2.7 ' L of EtOAc/heptane, after which 2.7L of heptane was added to the reactor.
  • the contents of the reactor were cooled to 25 ⁇ C and the resulting mixture was filtered under 5 psig nitrogen while washing with 4 L of heptane.
  • reaction mixture was transferred over a 2 hour period into another 22 L flask equipped with a mechanical stirrer, transfer line, vacuum line, and containing 4 L of 1:1 acetic acid; THF solution cooled in an ice- water bath.
  • the quenched solution was stirred for 30 minutes and then diluted with 4 L of 2M NH 4 OH in saturated aqueous NH 4 CI and 2 L of water.
  • the biphas ⁇ c mixture was stirred for 15 minutes and the phases separated.
  • the organic phase was washed 4x with 4 L aliquots ofthe2M NH4OH solution.
  • the resulting oil was taken up in 50 ml water and washed with 2x50 mL diethyl ether.
  • the aqueous layer was poured into 50 mL saturated sodium bicarbonate to give a solution of pH 7-8 (add more sodium bicarbonate if necessary) to give a white solid which was extracted with 4x50 mL diethyl ether.
  • the ether layers were combined and washed with b ⁇ ne, dried over anhydrous sodium sulfate, filtered into a flask containing 9.4 g (104 mmol) oxalic acid dissolved in 60 mL diethyl ether.
  • HEK 293 cells stably expressing recombinant porcine ⁇ 2 ⁇ -1 and human ⁇ 2 ⁇ -2 subu ⁇ ts were constructed previously (Gong KC. etal. (2001) J. Membr. Biol. 184:35-43) were grown under normal cell culture conditions (RPMI-1640 media with io% FBS, 200 ⁇ g G418, and 1% penicillin/ streptomycin at 37° C with 5% CO 2 ) until reaching 90% confluency in T-75 flasks, at which time they were harvested. Cells were suspended in ice-cold 5mM
  • Protein concentration was measured by well known methods.
  • the fH]-gabapentin SPA binding assay was performed in Costar 363296-well, clear bottom assay plates using wheat germ agglutinin coated polyvinyl toluene scintillation proximity assay (SPA) beads (Amersham Biosciences).
  • SPA polyvinyl toluene scintillation proximity assay
  • Alpha-2-delta-1 or o ⁇ -2 membrane proteins (10-20 ⁇ g protein per well) and SPA beads (0,5 mg per well) were mixed with 30 nM
  • mice Male, C57BL/6J mice (Jackson Laboratories, Bar Harbor, Maine) were used for vogel conflict, locomotor activity and accelerating rotarod testing. Animals were received at 5 weeks old and acclimated to the facility 1 week prior to testing. Male, DBA/2J mice 3 week old (Jackson Laboratories, Bar Harbor, Maine) were used for evaluating anticonvulsant activity, Mice were housed 5/isolator in a temperature/humidity-controlled room under a 12; 12 hour lighfcdark schedule (lights op at 6:00 AM) with food and water available ad libitum. AH procedures were carried out in compliance with the NlH Guide for the Care and Use of Laboratory Animals under a protocol approved by the PGRD Animal Use Committee.
  • the test apparatus consists of 12 modular operant chambers (Coulboum Instruments).
  • the front and back of the test chambers are made of clear Plexiglas.
  • the front doors are covered to reduce distractions from inside the test room.
  • the backs face a wail, away from the flow of traffic in the testing room and remain uncovered to provide the opportunity for observations.
  • All chambers have stainless steel grid floors and measure 7 x 7 x 12 inches.
  • Each test chamber is modified with an internal chamber made of clear Plexiglas measuring 6.75 x 3.5 x 1.5 inches.
  • the reduced chamber space limits the animal's activity and directs behavior towards the opening on the side of the chamber 1.5 inches above the floor.
  • a module optical lickometer mounted at the opening is used to measure licking.
  • a water bottle attaches to the outside of the cage and the drink tube extends into the opening through the module.
  • the reinforcer consists of a 1:1 mixture of evaporated milicwater.
  • a photo beam is piped across two glass rods that reside adjacent to the tip of the drink tube. Each time the animal licks the drink tube the beam is broken and licks are automatically recorded. Shock is delivered between the grid floor and the drink tube using a (Coulboum Instruments) programmable universal shocker for one second, but terminates Immediately when contact between the animal and drink tube is broken.
  • mice were placed upon a wire mesh, 4- inch square, suspended from a steel rod 12 to 18 inches from the top of a table. The square was slowly Inverted through 180 degrees and mice observed for 30 seconds. Any mouse falling from the wire mesh was scored ataxic.
  • Anticonvulsant testing started by placing individual mice into an enclosed acrylic plastic chamber (21-cm height, approximately 30-crn diameter) with a high-frequency speaker (4-cm diameter) in the center of the top lid, An audio signai generator (Protek Model B-810) was used to produce a continuous sinusoidal tone that was swept linearly in frequency between 8 and 16 kHz once each 10 milliseconds. The average sound pressure level duing stimulation was approximately 100 dB at the floor of the chamber. Mice were placed within the chamber and allowed to acclimatize for 1 minute.
  • DBA/2J mice in the vehicle-treated group responded to the sound stimulus (applied until tonic extension occurred, or for a maximum of 60 seconds) with a characteristic seizure sequence consisting of wild running followed by clonic seizures, and later by tonic extension, and finally by respiratory arrest and death in 90% or more of the mice.
  • a characteristic seizure sequence consisting of wild running followed by clonic seizures, and later by tonic extension, and finally by respiratory arrest and death in 90% or more of the mice.
  • vehicle-treated mice the entire sequence of seizures to respiratory arrest lasted approximately 10 to 15 seconds.
  • Locomotor activity (LMA) testing is performed using 16-Beam Digiscan Animal Activity Monitors (Acousca ⁇ Electronic, Colombus, OH). Each chamber consists of a Plexiglas box measuring 16 x 16 inches with a Plexiglas insert that divides the box into 4 equal quadrants 7.5 x 7,5 inches each. Two mice are placed in diagonal quadrants for testing with the lights on and infrared beams located on the perimeter of the chamber detect movement The entire test chamber is enclosed in a sound reduction chamber to reduce extraneous noise during the test session. Prior to the test day all mice are fasted overnight On the test day mice receive vehicle or test Compound 30 minutes prior to testing and total distance (cm) is recorded in 5-minute blocks for 1 hour, Data is reported as group Means + SEM
  • the test equipment consists of four programmable SmartRod® chambers (AccuScan Instruments, Columbus, OH). Each chamber is 36 cm (h) x 11 cm (w) x 30 (d) and equipped with a rotating rod horizontally affixed 32,5 cm on center above the grid floor. The rod spans the 11 cm width of the chamber and is 3 cm in diameter.
  • the equipment is programmed to accelerate the rod at a rate of 0.25 rev/sec to a maximum speed of 20 rpm over 62 seconds, after which the rod decelerates over the final 5 seconds to the stop position.
  • the total cycle time for one trial is 67 seconds. For each trial the mouse is suspended by the tail directly above the rod for three seconds simultaneous to the start of the cycle, then gently lowered onto the rotating rod.
  • Compound A and Compound B were each separately dissolved in sterile saline and each dose was delivered in a total volume of 5 ⁇ l_ at doses of 3 -30 ⁇ g/mouse.
  • Vehicle control mice received 5 ⁇ l_ of sterile saline.
  • a 27g steel needle calibrated to a depth of 3 mm was attached to a 25 ⁇ L Hamilton syringe for intraeerebroventricular (ICV) administration. Mice were manually restrained and the injection site was found by locating bregma on the skullcap with the tip of the needle. One mm to the lateral and 2 mm to the posterior of bregma the needle was dropped through the skull and into the ventricles and drug or vehicie was administered in 5 ⁇ L bolus.
  • ICV intraeerebroventricular
  • Compound A was found to displace [ 3 H]-gabapenti ⁇ binding with moderate-to-high affinity to membrane proteins prepared from recombinant porcine ⁇ 2 ⁇ -1 cells, whereas Compound A displaced [ 3 H]-gabapentin binding at very low affinity to membrane proteins prepared from recombinant human ⁇ 2 ⁇ -2 cells.
  • the K, values for displacement of binding of [ 3 H]-gabapenti ⁇ to recombinant porcine Ot 2 S-I and recombinant human ⁇ *&-1 cell membranes were 35 nWl, and 1670 ⁇ M, respectively, Selectivity for the ⁇ 2 ⁇ -1 is 48-fold greater than for the ⁇ 2 ⁇ -2 site, in comparison, the non-selective Compound (Compound B) displaced [ 3 H]- gabapentin in each of the membrane preparations with a Ki value of 32.8 nM for recombinant porcine ⁇ 2 ⁇ -1 and 34.9 nM for recombinant human ⁇ z ⁇ -2 cell membranes.
  • the Kj value determined for gabapentin was 75 nM for recombinant porcine a ⁇ - ⁇ , and 114 nM for recombinant human ⁇ 2 ⁇ -2 cell membranes (all values in this section are mean),
  • Compound A was compared to Compound B in the Vogel Conflict Test, sound- induced tonic seizures, and the locomotor activity and rotarod adverse effect models.
  • the effects of ICV dosing on C57BL/SJ mice following a 30-minute pre-treatme ⁇ t time using Compound A and Compound B in the Vogel Conflict test are shown in Tables 1 and 2, respectively.
  • the ⁇ 2 ⁇ -1 selective ligand, Compound A (330 ⁇ g/mouse), produced a dose dependent a ⁇ xiolytic-like effect in Vogel Conflict as reflected by an increase in mean shock episodes compared to the concurrent vehicle control group (Table 1).
  • the vehicle control group mean + SEWl was 10.8 + 1.1.
  • the minimally effective dose (MED) for Compound B was 3 ⁇ g/mouse with a magnitude of the response at this dose of 31,2 + 8.2. Therefore, both compounds were shown to produce an anxiolytiotike effect.
  • Compound A (3-30 ⁇ g/rnouse), produced a dose dependent increase in protecting DBA/2J mice from tonic seizures.
  • the ED50 value for Compound A was determined to be 6.6 ⁇ g ICV; with a S5% confidence interval of [4.2 to 10.3].
  • Compound B (3-30 ⁇ g/mouse) produced a dose dependent increase in protection of DBA/2J mice from tonic seizures.
  • the ED50 value for Compound B was determined to be 11.3 ⁇ g ICV; with a 95% confidence interval of [5.7 to 2ZT].
  • (percent) of one or more adverse effects following administration of an O 2 S-I selective ligand may be reduced by at least 10 percent or greater.
  • Compound A is an ⁇ 2 ⁇ -1 selective ligand.
  • Compound A prefers recombinant porcine O 2 S-I protein with binding inhibition Ki of 35 nM, whereas its binding inhibition Ki to recombinant human 0 3 6-2 protein was 48-fold less at 1670 nM.

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Abstract

The invention is directed to a method of treating a disorder or condition using an alpha-2-delta ligand with high affinity and selectivity for the alpha-2-delta-1 calcium channel subunit subtype. Administration of the alρha-2-delta-1 selective ligand, or a pharmaceutically acceptable salt, to a mammal, including a human, avoids or reduces the adverse effects compared to the administration of an alpha-2-delta non-selective ligand.

Description

METHODS OF TREATMENT USING ALPHA-2-DELTA-1 SELECTIVE COMPOUNDS
FIELD OF THE INVENTION
The invention is directed to methods of treating disorders or conditions by administering a selective ligand preferring alpha-2-de!ta-1 subunit of calcium channel membrane proteins. Such ligaπds are useful for treating CNS and pain-related disorders and conditions and may avoid or reduce adverse effects associated with administration of nonselective alpha-2-delta ligaπds.
BACKGROUND, OF THE INVENTION
Voltage-gated calcium channels are formed by combinations of the pore-forming alρha-1 (α,) subunit, and auxiliary alpha-2-delta, beta, and gamma (c-:_δ, β and γ, respectively) proteins (Catteraϋ W.A. (2000) Annul. Rev. Cell Dev. Biol. 16: 521-555). The oc2δ protein is known to regulate both the calcium channel density and voltage-dependent kinetics of these calcium channels (Felix R. ef a/. (1997) J, Neurαsci. 17: 6884-6891; Klugbauer N, etal. (1999) J. Neurosci. 19: 684-691; Hobom M. ef a/.(2000) Eur. J. Neurosci. 12:1217-1226; and Qϊn N, ef a/. (2002) MoI. Pharmacol. 62:485-496). The OG2S protein is encoded by a single gene and post-translationally cleaved to α2 and δ subunits. The α2 subunit is a highly glycosylated extracellular protein and is associated with the membrane anchor protein δ by disulfide linkage (Wang M, ef a/. (1999) Biochern. J. 342:313-320; Marais E. et a!. (2001) MoI. Pharmacol. 59:1243-1248; and Gong H.C, er a/. (2001) J. Mernbr. Biol. 164:35-43). Molecular cloning of the α2S subunits has revealed four α2δ subtypes in various species (Qin N. etal. (2002) MoI, Pharmacol. 62:485-496). See also
WO 00/20450, which refers to c-zδ-1, αzδ-2, α∑δ-3, and α2δ-4 as αaδ-A, O-2S-B1 ct2δ-C, and «28- D, respectively. The GenbanK accession numbers for α2δ-1 subunits include: M76559 (human); U73483, U734S4, U73485, U73486, and U73487 (mouse); M86S21 (rat); and AF077Θ65 (pig). See also Brown and N. Gee. (1998) J. Biol. Chem, 273:25458-25465. The accession numbers for α2δ-2 subunits include: AJ251368, AJ251367.1 , Abo11130.1 , NM-
006030.1, AF040709, AF042792, AF042793 (human); AF247139, NM-02063.2. AF247141, AB093246.1, AK0446D3.1 (mouse); and AF486277.1 and NM-175592.2 (rat).
Non-selective α≤S ligands bind with high affinity to two of the subtypes of calcium channel α2δ subtypes. For example, Gabapentiπ, 1-amiπomethyl-oyclohexyl-acetic acid (Neurontin®), is recognized as an anti-epileptic drug which binds with high affinity to αHδ-1 and α2δ-2. Additionally, Pregabslin, S-(+) 3-aminomethyl-5-methyl-hexanoic acid (Lyrica10), also binds with high affinity to the C-2S-I and α2δ-2 subtypes. Both Gabapeπtiπ and Pregabalin are coπsldered to be non-selective α2δ ligands (Gong H.C. ef a/. (2001) J, Membr. Biol. 184:35- 43). •
SUMMARY OF THE INVENTION 5
This invention relates to methods of treating disorders or conditions in a mammal, including a human, comprising administering to said mammal a therapeutically effective amount of an aIpha-2-delta-1 (α&δ-l) selective ligand, or a pharmaceutically acceptable salt thereof, wherein said disorder or condition is selected from pain, fibromyalgia, epilepsy, ] 0 restless legs syndrome, hot flashes, mood disorders and sleep disorders.
As shown below, the administration of an oaδ-i selective ligand avoids or reduces one or more adverse effects associated with the administration of an o^δ non-selective ligand, including but not limited to sedation, dizziness, ataxia, and asthenia. The avoidance or reduction of adverse effects following the administration of an α2δ-1 selective ligand is 15 attributed to the high binding affinity of the c-aδ-l selective ligand to the calcium channel subuπit cfeδ-1 subtype.
DETAILED DESCRIPTION OF THE INVENTION
20 The present invention is directed to methods of treating a disorders or conditions in a mammal, including a human, comprising administering to said mammal a therapeutically effective amount of an alpha-2-delta-1 (05.6-1) selective ligand, or a pharmaceutically acceptable salt thereof. Such ligands are useful far treating central nervous system (CNS) and pain-related disorders and conditions and may avoid or reduce adverse effects
25 associated with the administration of αgδ non-selective ligands. The CNS and pain-related disorders include, but are not limited to pain, fibromyalgia, epilepsy, restless legs syndrome, hot flashes, mood disorders, and steep disorders. The adverse effects include but are not limited to sedation, dizziness, ataxia, and asthenia.
The degree of binding to the O-2S subunit can be determined using the radioligand
30 binding assay using [3H]gabapentiπ and the oςδ subunit derived from porcine brain tissue, as described by N. S. Gee et a!., J. Biol. Client., 1996, 27^:5879-5776, An "α2δ-l selective" ligand means an O3S-I selective ligand, or pharmaceutically acceptable salt thereof, wherein said ligand or salt is at least 30-fold more effective in displacing bound [3H]-gabapentin from the αjδ-1 subunit than the αsδ-2 subunit; as determined by comparing the binding inhibition
35 value (Kj) of the compound for displacing [3H]-gabapeπtin bound to the α≥δ-i subunit, to the
. Kt of the ligand for displacing [3H]-gabapentin bound to the αjδ-2 subuπit. Thus, for the purposes of the invention, the term "selective" is in reference to the ration of Ki values of a ligand for displacing [aH]-gabapentin bound to the ααδ-1 subunit in comparison to the Ki value of a compound for displacing [3H]-gabapentiπ bound to the α2δ-2 subunit.
Methods for determining such K, values are known in the art, and also described herein. Therefore, Cx3S-I selective ligands can be identified among test compounds by comparing the K-, of the test Iigand for displacing [3H]-gabapentin bound to the α2δ-1 subunit, to the & of the compound for displacing [sH]-gabapentiπ bound to the a^b-i subunit.
For the purposes of the invention, one or more αjδ-1 selective ligands can be identified among α2δ ligands. Alpha-2-delta-1 ligands may include those compounds that are generally or specifically disclosed in the following references: US4024175, EP0641330, including 3-methylgabapeπtin; US5563175, WO97/33858, WO97/33859, WQ/99/31057,
WO99/31074, WO97/291Q1, WO02/0SS839, including [(1R,5R,6S)-6- (amiπomethy!)bicyclo[3,2.0]hept-6-yl]acetio acid; WO99/31075, including 3-{1-aminomethyl- cyclohexylmethyl)-4H-[1 ,2,4]oxadiazol-5-one and C-[1-(1H-tetrazo!-5-y)methyl)-cycloheptyl]- methylamine; WO99/21624, including (3S,4S)-(1-aminomethyi-3,4-dimethyl-oyclopenty[)- acetic acid; WO01/90Q52, WO01/28978, including (1α,3α,5α){3-amino-methyl- bicyc|o[3.2,0]hept-3-y|)-acetic acid; WOS8/17627, WO00/76958, including (35,SR)- 3-aminαmethyl-5-methyl-octanoio acid; WO03/0S2807, including (3S,5R}-3«arπiπo-5-rπethyI- heptaπoic acid, (SS.δRJ-S-amino-S-rnethyl-octaπαϊc acid; WO04/039367, including (2S,4S)-4- (34luorobenzyl)proline and (2S,4S)-4-(3-ohlorophenoxy) proline; EP1178034. EP12Q1240, W099/31074, WO03/000642, WO02/22568, WOQ2/30871, WO02/308S1, WO02/100392,
WO02/100347, WO02/42414, WO02/32736; WO02/28881; US 10/935,826 (publication 20050059654); US 10/640,515 (publication 200400092522; WO04/054566; US 10/401,060 (publication 20030195251) and US 10/949,908 (publication 20050124668); or pharmaceutically acceptable salts, esters or solvates thereof, all of which publications are hereby incorporated herein by reference in their entireties.
Alpha-2-delta ligands include those depicted by the following formula (I):
Figure imgf000004_0001
wherein X is a carboxylic acid or carboxylic acid bioisostere; n is 0, 1 or 2; and R1, R1a, R2, R2a, R3, R3a, RΔ and R4a are independently selected from H and C1-C6 aikyi, or R1 and R2, or R2 and R3, are taken together to form a C3-C7 cycloalkyl ring, which is optionally substituted with one or two substituents selected from C1-Ce aikyl, or a pharmaceutically acceptable salt or solvate thereof.
In formula (I), suitably, R1, R1a, R23, R3=, R4 and R4a are H and R2 and R3 are independently selected from H and methyl, or R1a, R2a, H35 and R4a are H and R1 and R2 or R2 and R3 are taken together to form a C3-Cj cycloalkyl ring, which is optionally substituted with one or two methyl substituents. A suitable carboxylic acid bioisostere fe selected from tetrazolyl and oxadiazoloπyl. X is preferably a carboxylic acid.
In formula (I), preferably, R1, R1a, R2a, R3*, R4 and R43 are H and R2 and R3 are independently selected from H and methyl, or R1a, R2a, R3a and R4a are H and R1 and R2 or R2 and R3 are taken together to form a C4-C5 cycloalkyl ring, or, when π is 0, R1, R1a, R2B, R3a, R4 and R"3 are H and R2 and R3form a cydopentyl ring, or, when n is 1, R1, R1a, R2a, R33, R4 and R*1 are H and R2 and R3 are both methyl or R1, R1a, R2*, R3a, R4 and R4a are H and R2 and R3 form a cyclobutyl ring, or, when n is 2, R1, R1a, R2, R23, R3, R3a, R4 and R"*9 are H, or, π is 0, R1, R1a, R2a, R3a, R" and R4a are H and R2 and R3form a cydopentyl ring.
Alpha2delta ligands also include those depicted by the following formula (II):
Figure imgf000005_0001
(H) wherein: n is 0 or 1, Rn is hydrogen or C1-C6 alkyl; R2 is hydrogen or CrC6 aikyl; R3 is hydrogen gr C1-Ce aikyl; R4 is hydrogen or C1-C5 alkyl; Rs is hydrogen or CrC6 alkyl and R2 is hydrogen or C1-Ca alkyl, or a pharmaceutically acceptable salt, ester or solvate thereof.
In one embodiment, 0-26 ligands are Compounds of formula (II) wherein R1 is Ci-C8 alkyl, R2 is methyl, R3 - R8 are hydrogen and π is 0 or 1, In another, O2S ligands are Compounds of formula (H) wherein R1 is methyl, ethyl, n-propyl or n-butyl, R2 is methyl, R9 -
R6 are hydrogen and n is 0 or 1. When R2 is methyl, R3 — R8 are hydrogen and n is 0, R1 is suitably ethyl, n-propyl or π-butyl. When R2 is methyl, R3 - R6 are hydrogen and n is 1 , R1 is suitably methyl or n-propyl. Compounds of formula (II) are suitably in the 3S, 5R configuration. Alρha-2-deIta ligands also include: f(1R,5R,6S)-6-(amiπomethyl)biαycle-[3.2.0]hept-6- yl]acetic acid, 3-(1-aminomethyl-cyclohexylmethyl)-4H-[1,2,4]oxadlazol-5-one, C-[1-(1H- tetrazoI-5-ylrnethyl)-cydoheptyl]-methylamiπe, (3S,4S)-(1-amJπornethyl-3.4-dimethyl- cyclopentyl)-acetig acid, (1α,3αI5α)(3-amiπo-methy!-bicyclo[3.2ιO]hβpt-3-yl)-acetic acid, (3S,5R)-3-aminomethyl'5-methyl-octaπoϊe acid, (3S,5R)-3-amino-5-methyI-heptanoic acid, (SS.SRJ-S-amino-S-methyl-nonanoic acid, (SS.δRJ-S-aminα-S-rnethyl-octanoic acid, {2S,4S)^t- S-
(3-chlorophenoxy) proline and (2S,4S)-4-(3-fluorobenzyl) proline and the pharmaceutically acceptable salts and solvates thereof, For the purposes of the present invention, cfcS nonselective ligands include gabapentiπ (3S, 5R)-3-amiπσmethyI-6-cydopropyl-5-methyl- hexanoic acid, and pregabalin, S-(+) eπantiomer of 3-aminomethyl-S-methyl-hexanoic acid (Lyrica®). For purposes of the present invention, an Ot3S-I selective ligand, (3S,4R,5R)-3- Arnino-4,5-dimethyloc!tanoic acid, hereinafter "Compound A" i$ described.
Figure imgf000006_0001
Compound A
Selectivity is determined by calculating the binding affinity Ki value from the amount °f [3H]-gabapentin bound to calcium channel aφ subuπit membrane proteins prepared from both recombinant porcine (X2S-I cells and recombinant human α2δ-2 cells. The K1 values for displacement of binding of [3H]-gabapentin to recombinant porcine α2δ-l and recombinant human a.φ-2 cell membranes correspond to the binding affinity of the Qt2S ligands. The ratio of the respective K| values of the two recombinant cell membrane proteins corresponds to binding selectivity toward ctaS-1. Selectivity of an oc2δ-1 selective ligand to the O8M subtype is at least 30-, 40-, 50-, 100-, 500-, or 1000-fold more than for binding to the αaδ-2 subunit The mean K1 values for Compound A are 35 nM and 1670 nM for recombinant porcine αjδ-1 and recombinant human α2δ-2 cell membranes, respectively, accounting for a 48-fold selectivity for the Ct2S-I subunit
All that is required to practice the method is to administer an αaδ-1 selective ligand, or a pharmaceutically acceptable salt thereof, in an amount that is therapeutically effective to treat one or more of the disorders or conditions described herein. Such therapeutically effective amount will generally be from about 1 to about 300 mg/kg of subject body weight
Typical doses will be from about 10 to about 5000 mg/day for an adult subject of normal weight In a clinical setting, regulatory agencies such as, for example, the Food and Drug Administration ("FDA") in the US, may require a particular therapeutically effective amount In determining what constitutes an effective amount or a therapeutically effective amount of an α3δ-1 selective iigand, or a pharmaceutically acceptable salt thereof, fortreating one or more of the disorders or conditions referred to herein according to the invention method, a number of factors will generally be considered by the medical practitioner or veterinarian in view of the experience of the medical practitioner or veterinarian, published clinical studies, the subject's age, sex, weight and genera! condition, as well as the type and extent of the disorder or condition being treated, and the use of other medications, if any, by the subject. As such, the administered dose rnay fall within the ranges or concentrations recited above, or may vary outside, i.e., either below or above, those ranges depending upon the requirements of the individual subject, the severity of the condition being treated, and the particular therapeutic formulation being employed, Determination of a proper dose for a particular situation is within the skill of the medical or veterinary arts, Generally, treatment may be initiated using smaller dosages of the Ct2S-I iigand that are less than optimum for a particular subject. Thereafter, the dosage can be increased by small increments until the optimum effect under the circumstance is reached. For convenience, the total daily dosage may be divided and administered in portions during the day, if desired.
A preferred embodiment relates to a method of treating a disorder or condition selected from the group consisting of pain such as acute pain, chronic pain, pain resulting from soft tissue and peripheral damage such as acute trauma; complex regional pain syndrome also referred to as reflex sympathetic dystrophy, postherpetic neuralgia, occipital neuralgia, trigeminal neuralgia, segmental or intercostal neuralgia and other neuralgias; musculoskeletal pain such as pain associated with strains, sprains and trauma such as broken bones; central nervous system pain such as pain associated with spinal pain, spinal pain associated wfth cord or brain stem damage; lower back pain, sciatica, dental pain, myofascial pain syndromes, episiotomy pain, gout pain, and pains resulting from burns; deep and visceral pain such as heart pain, musde pain, eye pain, inflammatory pain, orofascial pain, for example odontalgia; abdominal pain, and gynecological pain, for example, dysmenorrhea, labor pain and pain associated with endometriosis; somatogenic pain; pain associated with nerve and root damage, such as pain associated with peripheral nerve disorders, for example, nerve entrapment, brachial plexus avulsions, and peripheral neuropathies; pain associated w'rth limb amputation, tic douioureaux, neuroma, or vasculitis; diabetic neuropathy, ehemo-therapy-induced neuropathy, acute herpetic and postherpetic neuralgia; atypical fascial pain and arachnoiditis; occipital neuralgia, allodynia, hyperalgesia, idiopathic pain, pain associated with restless legs syndrome, pain associated with gallstones, neuropathic and non-neuropathic pain associated with carcinoma, phantom limb pain, temperomaπdibular pain and maxillary sinus pain, pain associated with gastrointestinal disorders, for example, ulcerative colitis, dyspepsia, proctitis, chronic pancreatitis; post operative pain, scar pain, and non-neuropathic pain such as pain associated with arthralgia, myalgia, vasculitis, and fibromyalgia in a mammal, comprising administering to a mammal in need of such treatment a therapeutically effective amount of an oφ-Λ ligand, or a pharmaceutically acceptable salt thereof.
Another preferred embodiment relates to a method of treating a disorder or condition selected from the group consisting c-f mood disorders, such as depression, or more particularly, depressive disorders, for example, single episodic or recurrent major depressive disorder, severe unipolar recurrent major depressive episodes, and melancholic depression; seasonal affective disorder, conduct disorder and disruptive behavior disorder, obsessive compulsive disorder, stress related somatic disorders and anxiety disorders, for example, generalized anxiety disorder, social anxiety disorder; stress disorders including post traumatic stress disorder and acute stress disorder in a mammal, comprising administering to a mammal in need of such treatment a therapeutically effective amount of an O2S-I ligand, or a pharmaceutically acceptable salt thereof. See Diagnostic and Statistical manual of Mental Disαrtders, Fourth Edition (DSM-IV)1 American Psychiatric Association, Washington, D. C, May 1194, pp. 435-436.
Another preferred embodiment relates to a method of treating a disorder or condition selected from the group consisting of sleep disorders, for example, insomnia {e.g., primary insomnia including psychophysiological and idiopathic insomnia, secondary insomnia including insomnia secondary to restless legs syndrome, insomnia secondary to anxiety, insomnia secondary to fibromyalgia, insomnia secondary to pain and neuropathic pain, and transient insomnia), sleep deprivation, REM sleep disorders, sleep apnea, hypersomnia, parasomnras, sleep-wake cycle disorders, jet lag, narcolepsy, sleep disorders associated with shiftwork or irregular work schedules, deficient sleep quality due to a decrease in slow wave sleep caused by medications or other sources, and other sleep disorders in a mammal in need of such treatment a therapeutically effective amount of an Qt2S-I ligand, or a pharmaceutically acceptable salt thereof. Another preferred embodiment relates to a method of treating a disorder or condition selected from the group consisting of fibromyalgia (FIVl). Fibromyalgia is a chronic syndrome characterized mainly by widespread pain, un-refreshing sleep, disturbed mood, and fatigue. Syndromes commonly associated with FWl include irritable bowel syndrome, and migraine headaches, among others. Success of treating FWl with a single pharmacological agent has been characterized as modest and results of clinical trials have been characterized as disappointing. It is believed that based on current understanding of the mechanisms and pathways involved in FM, multiple agents will be required, aimed at the major symptoms of pain, disturbed sleep, mood disturbances, and fatigue. Fibromyalgia patients are often sensitive to side effects of medications, a characteristic perhaps related to the pathophysiology of this disorder (Barkhuizen A, Rational and Targeted pharmacologic treatment of fibromyalgia Rheum Dis Clin N Am 2002; 28: 261-290; Leventhal LJ. Management of fibromyalgia, Ann intern Med 1999;131:850-S).
While FM is a complex disorder with multiple facets, this complexity can be well assessed (Yunus MB, A comprehensive medical evaluation of patients with fibromyalgia syndrome, Rheum Dis N Am 2002; 28:201-217). The diagnosis of FM is usually based on the
1990 recommendations of the American College of Rheumatology classification criteria (Bennett RM, The rational management of fibromyalgia patients. Rheum Dis Clin N Am 2002; 28: 161-199; Wolfe F, Smythe HA, Yunus MB, Bennett RM, Bombardier C, Goldenberg DL, et al. The American College of Rheumatology 1990 criteria for the classification of fibromyalgia: Report of the Multicenter Criteria Committee. Arthritis Rheum 1990; 33:160-72). Evaluation, management, and pharmacological treatment of fibromyalgia have been reviewed (Barkhuizeπ A, Rational and Targeted pharmacologic treatment of fibromyalgia. Rheum Dis Clin N Am 2002; Buskila D, Fϊbomyalgia, chronic fatigue syndrome and myofaoial pain syndrome. Current opinions in Rheumatology 2001; 13: 117-127; Leventhal LJ.
Management of fibromyalgia. Ann Intern Med 1999;131:850-8; Bennett RM, The rational management of fibromyalgia patients. Rheum Dis. Clin N Am 2002; 2δ: 181-199; Yuπus MB, A comprehensive medical evaluation of patients with fibromyalgia syndrome, Rheum Dis N Am 2002; 28:201-217). Another preferred embodiment relates to a method of treating a disorder or condition selected from the group consisting of epilepsy. Epilepsy is an episodic disturbance of consciousness during which generalized convulsions may occur. The condition is of unknown etiology, often hereditary, and is manifested by symptoms of a peculiar sensation, smell or feeling, called an "aura" proceeding the loss of consciousness and often convulsions. The term "epilepsy" is a collective designation for a group of chronic central nervous system disorders having in common brief episodes (seizures) associated with loss or disturbance of consciousness, usually but not always with characteristic body movements (convulsions) and sometimes autonomic hyperactivity, and always correlated with abnormal and excessive EKG disturbances. Epileptic seizures in a mammal, including a human, are classified on the basis of the clinical manifestation© of the onset and the EKG pattern. Sei∑ure types are often used to classify the particular type or types of epilepsy which include gran mal (generalized tσnic- chronic), absence (petit mal), cortical focal, temporal lobe (psychomotor), and infantile that often occur in adolescents.
Another preferred embodiment relates to a method of treating a disorder or condition selected from the group consisting of restless legs syndrome (RLS). Restless legs syndrome is a common, potentially disabling condition that affects about 10% to 15% of the general population and yet is often unrecognized and misdiagnosed. It is mainly diagnosed clinically and only rarely requires polysomnography. The condition is usually primary and treatable. Restless legs syndrome is a chronic condition. Symptoms may worsen with age, and the most disabling feature is sleep onset insomnia. Restless legs syndrome is a sensory-motor
(movement) disorder characterized by uncomfortable sensations in the legs, which are worse during periods of inactivity or rest while sitting or lying down. Individuals affected with RLS describe the sensations as pulling, drawing, crawling, tingling, pins and needles, and sometimes painful sensations that are usually accompanied by an overwhelming urge to move the legs. Sudden muscle jerks may also occur. Movement provides temporary relief from the discomfort In rare cases, the arms may also be affected. Symptoms may also interfere with sleep onset (sleep onset insomnia). A variety of etiologies have been proposed for RLS including pregnancy, polyneuropathy, and drug withdrawal. However, the common theme appears to be Inflammation in the central nervous system associated with impaired blood flow to spinal nerve roots, Changes in nerve conduction have also been reported, suggesting abnormalities in spinal cord function.
Another preferred embodiment relates to a method of treating a disorder or condition selected from the group consisting of hot flashes, Hot flashes occur in both male and females mammals, including humans. Females having a low level of estrogen are prone to suffer from hot flashes. This deficiency can be due to radiation therapy, which can prematurely induce menopause, or pan be caused by specific medications such as anti-estrogen treatment or certain drugs (e.g. Tamoxifen (Nolvadex)). Further, hot flashes may be secondary to menopause or postmenopause, medical treatment, and cancer. In men, androgen deprivation can be a cause of hot flashes. Similarly, hormone imbalance can be drug-induced
(e,g. Lupron (Leuprolide) and Zoladex (Goserelin)) or radiation-induced. Surgery such as bilateral orchiectomy for prostate cancer or testicular cancer may also cause hot flashes in males.
The foregoing methods are also referred to herein, collectively, as "the invention methods".
Another preferred embodiment relates to a method of treating a disorder or condition selected from the invention methods, comprising administration of a therapeutically effective amount of an α2δ-1 ligand, or a pharmaceutically acceptable salt thereof, to a mammal, including a human, wherein at least one adverse effect is avoided or reduced. Another embodiment relates to a method of neuroprotection for brain damage caused by stroke, cardiac arrest, Alzheimer's disease, and other related conditions.
Another embodiment relates to any of the above methods wherein the «5,6-1 ligand, or a pharmaceutically acceptable salt thereof, is administered to a mammal, including a human, for the treatment of any two or more comorbid disorders and conditions referred to in any of the above methods.
Another embodiment relates to any of the above methods for treating any of the invention methods, comprising administration of an αjδ-1 selective ligand, wherein the said ligand is (3S,4R,5R)-3-Amino-4,5-dimethyloctaπoic acid or a pharmaceutically acceptable salt thereof, to a mammal, including a human. Another embodiment relates to adverse effects being avoided or reduced in a mammal, including a human, comprising the administration of an α.aδ-1 selective ligand, or pharmaceutically acceptable salt thereof. The term "adverse effects" includes, but is not limited to: dizziness, asthenia, ataxia, sedation, somnolence, fatigue, nystagmus, weight gain, ernesis, peripheral edema, dyspepsia, tremor, nervousness, amnesia, depression, twitching, myalgia, rhinitis, diplopia, amblyopia, malaise, hypertension, flatulence, purpura most often described as bruises resulting from physical trauma, arthralgia, vertigo, hyperkinesia, decreased or absent reflexes, increased reflexes, hostility, and abnormal vision. The term "avoid", "avoids", or "avoided" as used herein means that the adverse effect does not occur in the mammal following administration of an o.jδ-1 selective ligaπd, The term "reduce", "reduces", or "reduced" as used herein means that the frequency or incidence of one or more adverse effects has been decreased by at least 10 percent. Another embodiment relates to the administration of an Os2S-I selective ligand in an amount of about 10 to 5000 mg per day.
Another embodiment relates to the administration of an Cx2S-I selective ligand that is in a liquid or solid dosage form. Administration of a dosage form comprising an αaδ-1 selective ligand. or a pharmaceutically acceptable salt thereof, may be administered orally, parenteral^, subcutaneously, intravenously, intramuscularly, intraperitoneally, intracerebroventricularly, by intranasal instillation, by implantation, by intracavitary or intravesical instillation, iπtraocularly, intraarterially, iπtralesionaliy, intracerebroventricular, or by application to dermal, sublingual, or mucous membranes.
Another embodiment relates to a pharmaceutical composition comprising an O2S-I selective ligand and a pharmaceutically acceptable carrier. The term "pharmaceutically acceptable" refers to those properties and/or substances which are acceptable to the mammal, including a human, from a pharmacological and toxicological perspective regarding composition, formulation, stability, safety, and bioavailability,
Pharmaceutical compositions of an O2S-I ligand, or a pharmaceutically acceptable salt thereof, are produced by formulating the active compound in dosage unit form with a pharmaceutical carrier. Some examples of dosage unit forms are tablets, capsules, pills, powders, aqueous and nonaqueous oral solutions and suspensions, transdermals, and parenteral solutions packaged in containers containing either one or some larger number of dosage units and capable of being subdivided into individual doses. Some examples of suitable pharmaceutical carriers, including pharmaceutical diluents, are gelatin capsules; sugars such as lactose and sucrose; starches such as corn starch and potato starch; cellulose derivatives such as sodium carboxymethyl cellulose, ethyl cellulose, methyl cellulose, and cellulose acetate phthaiate; gelatin; talc; stearic acid; magnesium stearate; vegetable oils such as peanut oil, cottonseed oil, sesame oil, olive oil, corn oil, and oil of theobroma; propylene glycol, glycerin; sorbitol; polyethylene glycol; water; agar; alginic acid; isotonic saline, and phosphate buffer solutions; as well as other compatible substances normally used in pharmaceutical formulations,
The compositions to be employed in the invention can also contain other components such as coloring agents, flavoring agents, and/or preservatives. These materials, if present, are usually used in relatively small amounts. The compositions can, if desired, also contain other therapeutic agents commonly employed to treat the disorder or condition being treated. The percentage of the active ingredients in the foregoing compositions can be varied within wide limits, but for practical purposes it is preferably present in a concentration of at least 10% in a solid composition and at least 2% in a primary liquid composition. The most εatisfactory compositions are those in which a much higher proportion of the active ingredient is present, for example, up to about 95%.
The OC2S-I ligand, or a pharmaceutically acceptable salt thereof, may be administered in any form. Preferably, administration is in unit dosage form, A unit dosage form of the α2δ-l ϋgaπd, or a pharmaceutically acceptable salt thereof, to be used in this invention may also comprise other compounds useful in the therapy of the disorder or condition for which the α2δ- 1 ligand is being administered or a disorder or condition that is secondary to the disorder or treatment for which the αaδ-1 iigaπd is being administered.
Some of the compounds utilized in a method of the present invention are capable of further forming pharmaceutically acceptable salts, including, but not limited to, acid addition and/or base salts. The acid addition salts are formed from basic compounds, whereas the base addition salts are formed from acidic compounds. AIi of these forms are within the scope of the compounds useful in the method of the present invention,
Pharmaceutically acceptable acid addition salts of the basic compounds useful in the method of the present invention include nontoxic salts derived from inorganic acids such as hydrochloric, nitric, phosphoric, sulfuric, hydrαbromϊc, hydroiodic, hydrofluoric, phosphorous, and the like, as well nontoxic salts derived from organic acids, such as aliphatic mono- and dicarboxylic acids, phenyl-substituted alkartoic acids, hydroxy alkanoio acids, alkanedioic acids, aromatic acids, aliphatic and aromatic sulfonic acids, etc. Such salts thus include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, mαπohydrogenphosphate, dihydrogenphosphate, rπetaphosphste, pyrophosphate, chloride, bromide, iodide, acetate, trifluoroacetate, propionate, capryiate, isobutyrate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, mandelate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, phthalate, benzenesulfσnate, tαlueπesulfonate, phenylacetate, citrate, lactate, malate, tartrate, methanesulfoπate, and the like. Also contemplated are salts of amino acids such as arginate and the like and gluconate, galacturonate (see, for example, Berge S. Wl, et a!., "Pharmaceutical Salts," J, otPharma. ScL, 1977;66:1).
An acid addition salt of a basic compound useful in the method of the present invention is prepared by contacting the free base form of the compound with a sufficient amount of a desired acid to produce a nontoxic salt in the conventional manner. The free base form of the compound may be regenerated by contacting the acid addition salt so formed with a base, and isolating the free base form of the compound in the conventional manner, The free base forms of compounds prepared according to a process of the present invention differ from their respective acid addition salt forms somewhat in certain physical properties such as solubility, crystal structure, hygroscopicity, and the like, but otherwise free base forms of the compounds and their respective acid addition salt forms are equivalent for purposes of the present invention.
A pharmaceutically acceptable base addition salt of an acidic compound useful in the method of the present invention may be prepared by contacting the free acid form of the compound with a nontoxic metal cation such as an alkali or alkaline earth metal cation, or an amine, especially an organic amine. Examples of suitable metal cations include sodium cation
(Na+), potassium cation (K+), magnesium cation (Mg2+), calcium cation (Ca2+), and the like. EΞxamples of suitable amines are N.N'-dibenzylethyleπediamine, chloroprocaiπe, choline, diethanolamine, dicyclohexylamine, ethylenediamiπe, N-methylglucamine, and procaine (see, for example, Berge, supra., 1977).
A base addition salt of an acidic compound useful in the method of the present invention may be prepared by contacting the free acid form of the compound with a sufficient amount of a desired base to produce the salt in the conventional manner. The free acid form of the compound may be regenerated by contacting the salt form so formed with an acid, and isolating the free acid of the compound in the conventional manner. The free acid forms of the compounds useful in the method of the present invention differ from their respective salt forms somewhat in certain physical properties such as solubility, crystal structure, hygroscσpϊcity, and the like, but otherwise the saits are equivalent to their respective free acid for purposes of the present invention.
Certain of the compounds useful in the method of the present invention can exist in unsolvated forms as well as solvated forms, including hydrated forms. In general, the solvated forms, including hydrated forms, are equivalent to unsolvated forms and are intended to be encompassed within the scope of the present invention. Certain of the compounds useful in the method of the present invention possess one or more stereoceπters, and each center may exist in the R or S configuration. A method of the present invention may utilize any diastereomeric, enantiomeric, or epimeric form of an α2δ-1 ligand, or a pharmaceutically acceptable salt thereof, as well as mixtures thereof,
Additionally, certain compounds useful in the method of the present invention may exist as geometric isomers such as the entgegen (E) and zusammen (Z) isomers of alkeπyl groups. A method of the present invention may utilize any cis, trans, syn, anti, entgegen (E), or zusammen (Z) isomer of an αjδ-1 ligand, or a pharmaceutically acceptable salt thereof, as well as mixtures thereof.
Certain compounds useful in the method of the present invention can exist as two or more tautomeric forms. Tautomeric forms of the compounds may interchange, for example, via enolϊzatioπ/de-eπolization and the like. A method of the present invention may utilize any tautomeric form of an α2δ-1 ligand, or a pharmaceutically acceptable salt thereof, as well as mixtures thereof. For preparing pharmaceutical compositions from the ligands of the present invention, pharmaceutically acceptable carriers can be either solid or liquid. Solid form preparations include powders, tablets, pills, capsules, cachets, suppositories, and dispersible granules. A solid carrier can be one or more substances wnich may also act as diluents, flavoring agents, binders, preservatives, tablet disintegrating agents, or an encapsulating material.
In powders, the carrier is a finely divided solid which is in a mixture with the finely divided active component.
In tablets, the active component is mixed with the carrier having the necessary binding properties in suitable proportions and compacted in the shape and size desired. The powders and tablets preferably contain from five or ten to about seventy percent of the active compound, Suitable carriers are magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, tragacaπth, methyleeltulose, sodium carboκymethylcellulose, a low melting wax, cocoa butter, and the like. The term "preparation" is Intended to include the formulation of the active compound with encapsulating material as a carrier providing a capsule in which the active component, with or without other carriers, is surrounded by a carrier, which is thus in association with it. Similarly, cachets and lozenges are included. Tablets, powders, capsules, pills, cachets, and lozenges can be used as solid dosage forms suitable for oral administration.
For preparing suppositories, a low melting wax, such as a mixture of fatty acid glycerides or cocoa butter, is first melted, and the active component is dispersed homogeneously therein, as by stirring. The molten homogenous mixture is then poured into convenient sized molds, allowed to cool, and thereby to solidify.
Liquid form preparations include solutions, suspensions, and emulsions, for example, water or water propylene glycol solutions. For parenteral injection liquid preparations can be formulated in solution in aqueous polyethylene glycol solution.
Aqueous solutions suitable for oral use can be prepared by dissolving the active component in water and adding suitable colorants, flavors, stabilizing and thickening agents as desired.
Aqueous suspensions suitable for oral use can be made by dispersing the finely divided active component in water with viscous material, such as natural or synthetic gums, resins, methylcellulose, sodium carboxymethyleellulose, and other well-known suspending agents.
Also included are solid form preparations which are intended to be converted, shortly before use, to liquid form preparations for oral administration. Such liquid forms include solutions, suspensions, and emulsions. These preparations may contain, in addition to the active component, colorants, flavors, stabilizers, buffers, artificial and natural sweeteners, dispersants, thickeners, solubilizing agents, and the like.
The pharmaceutical preparation is preferably in unit dosage form. In such form the preparation is subdivided into unit doses containing appropriate quantities of the active -J 4-
compoπent. The unit dosage form can be a packaged preparation, the package containing discrete quantities of preparation, such as packeted tablets, capsules, and powders In vials or ampoules. Also, the unit dosage form can be a capsule, tablet, cachet, or lozenge itself, or it can be the appropriate number of any of these in packaged form. The quantity of active component in a unit dose preparation may be varied or adjusted from 0.1 mg to 1 g according to the particular application and the potency of the active component In medical use the drug may be administered three times daily as, for example, capsules of 100 or 300 mg. The composition can, if desired, also contain other compatible therapeutic agents. In therapeutic use, the compounds utilized in the pharmaceutical method of this invention are administered at the initial dosage of about 0.01 mg to about 100 mg/kg daily. A daily dose range of about 0.01 mg to about 100 mg/kg is preferred. The dosages, however, may be varied depending upon the requirements of the patient, the severity of the condition being treated, and the compound being employed. Determination of the proper dosage for a particular situation is within the skill of the art. Generally, treatment is initiated with smaller dosages which are less than the optimum dose of the compound. Thereafter, the dosage is increased by small increments until the optimum effect under the circumstances is reached. For convenience, the total daily dosage may be divided and administered in portions during the day, if desired. The term "treating", as used herein, refers to reversing, alleviating, inhibiting the progress of, or preventing the disorder or condition to which such term applies, or preventing one or more symptoms of such condition or disorder. The term "treatment", as used herein, refers to the act of treating, as "treating" is defined immediately above.
It should be appreciated that the terms "uses", "utilizes", and "employs" are used interchangeably when describing an embodiment of the present invention.
The term "therapeutically effective", as used herein, refers to the treatment of a mammal, including a human, with an amount of an α2δ-1 ϋgand, or a pharmaceutically acceptable salt thereof, to treat one or more of the invention disorders or conditions as described herein. The phrase "lower alkyl" means a straight or branched alkyl group or radical having from 1 to 6 carbon atoms, and includes methyl, ethyl, n-propyl, /-propyl, π-butyl, /-butyl, sec- butyl, ferf-butyl, π-pentyl, n-hexyl, and the like.
The term "alky!", as used herein, unless otherwise indicated, includes saturated monovalent hydrocarbon radicals having straight, branched or cyclic moieties or combinations thereof. Examples of "alkyl" groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, iso- sec- and tert-butyl, pentyl, hexyl, heptyl, 3-ethylbutyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohβxyl, cycloheptyl, norbornyl, and the like, The cycloalkyl groups are saturated monovalent carbocyclic groups containing from 3 to 7 carbons and are selected from cycloprαpyl, cyclobutyl, cyclopeπtyl, cyclohexyl, and cycloheptyi, unless otherwise stated.
In certain embodiments, this invention relates to methods of treatment utilizing the O2S-I selective ligaπd, {3$,4R,5R)-3-Amino-4,5-dirnetrιyloctanoic acid (hereinafter
"Compound A"), or a pharmaceutically acceptable salt thereof; which compound can be prepared by:
(R)-3-((R)-3-Methyl-riexaπoyl}-4-prienyl-oxazθlϊdiπ-2-oπe To the copper (1) bromide dimethylsulfjde complex (13.34 gτ 64,87 mmo!) in dry tetrahydrofuran (THF) (150 rnL) at -30°C under Nitrogen was added a 2M ether solution of Propylmagnesiurnchloride (64.87 rnL, 129.7 mrnol) and stirred for 20 minutes. (R)-3-But-2- eπoyl-4-pheπyl-oxa2olidin-2-oπe (15.0 g, 64.87 mmαl) in THF (60 ml_) was added over a 15 minute period at -35αC and let slowly warm up to room temperature over 4 hours. The mixture was cooled to 0*C and quenched with saturated ammonium chlc-ride solution. The suspension was extracted into ether, washed with 5% ammonium hydroxide solution and then with brine and dried over MgSO4. The solution was concentrated under reduced pressure to afford the titled Compound (13.34 g; 100% yield) as a white solid: 1H NMR (400 MHz, CDCI3) δ ppm Q.β (m, 6 H) 1.2 (m, 3 H) 1.6 (s, 1 H) 2.0 (m, 1 H) 2.7 (dd, »/=16.1, 8.5 Hs1 1 H) 3.0 (dd, J=15.9, 5.4 Hz1 1 H) 4.3 (dd, J=8.9, 3.8 Hz, 1 H) 4.7 (t, ./=8.3 Hz1 1 H) 5.4 (dd, J=8.8, 3.9 Hz1
1 H) 5.4 (dd, J=8.Q, 3.9 Hz, 1 H) 7.3 (m, 5 H). MS, rn/z (relative intensity): 276 [M+1H, 100%].
(R)~3-((2R,3R)-2,3-DimethyI-hexaπoyl)-4-phenyl-oχazolidin-2-one
To a 1M THF solution of Sodium hexamethyldisylamide (1S.2 g, 88.3 mmoi) at -7SDC was added via cannula a 00C solution of (R)-3-((R)-3-methyl4iexanoy!)-4-pheπyl-oxazolidin-2- one (18.7 g, 67.9 mmol) in 70 mL dry THF. The resulting solution was stirred at -780C for 30 minutes. Methyl iodide (48.2 g, 339.5 mmol) was added and stirring at -785C was continued for 4 hours. The reaction was quenched with saturated ammonium chloride solution, extracted into CHgCi2 and washed with 1M sodium Bisulfite. The solution was dried over MgSO4, concentrated and chromatographed in 10% ethylacetate in hexane to give the titled
Compound (11.1 g, 56.5% yield) as an oil. MS, m/z (relative intensity); 1H NMR (400 MHz, CDCI3) 6 ppm 0.8 (t, J=7.0 Hz, 3 H) 0.9 (d, J=6.6 Hz, 3 H) 1.0 (d, J=6.S Hz, 3 H) 1,0 (d, J=8.5 Hz, 1 H) 1.1 (m, 1 H) 1.4 (m, 1 H) 1.7 (m, 1 H) 3.7 (m, 1 H) 4.2 (dd, J=8.S, 3.4 Hz, 1 H) 4.6 (t, J=BJ Hz1 1 H) 5,4 (dd, J=8.7, 3.3 Hz, 1 H) 7.2 (m, 2 H) 7.3 (m, 3 H). MS, m/z (relative intensity):290 fM+1H, 100%]. (4S,5R)-4,5-Diphenyl-oxazolidϊn-2-one
To a 5 L round bottom flask equipped wϊth an overhead stirrer, thermocouple and distillation head, was charged 550 g (2.579 mol) of (IR^S^diphenyl-^-aminoethanol, 457 g (3.868 mol, 1.5eq) of diethyicarbonate, 18 g (0.258 mol, 0.1 eq) of NsOEt in 100 ml_ of EtOH and 3.5 L of toluene. The reaction was heated until an internal temperature of 90αC was reached and EtOH distillation began. The reaction was refluxed until an internal temperature of 110aC was reached (7 hours). For every 500 mL of solvent that was removed via the distillation head, 500 mL of toluene was added back to the reaction. A total of about 1.6 L of solvent was removed. The reaction was allowed to cool to room temperature and then filtered on a 3 L coarse fritted funnel with 2 psig N2. Nitrogen was blown over the cake overnight to give 580 g (94% yield) of the titled Compound: 1H NMR (DMSO) 7.090-6.985 (m, 6H), 6.930- 6.877 (m, 4H)1 5.900 (d, 1H, J = 8.301), 5.206 (d, 1H, J = 8.301).
(4S,5R)-3-((E}-2-Methyl-hex-2-enoyl)-4,5-dlphenyl-ox<3zolIdin-2-one (Alternative A) A 20 L jacketed reactor was fitted with a reflux condenser. To the reactor was charged 1100 g (4.597 mol) of (4S,5R)-4,5-diphenyl-oxazσlidin-2-one, 884 g (6.896 mot) (E)- 2-ιnethyl-2-peπtenαic acid, 1705 g (6.896 mol) of EEDQ, 48 g (1 ,149 rnol) of LiCI and 16 L of EtOAc, The reaction mixture was heated to 65 0C and was held for 200 minutes. The reaction mixture was cooled to room temperature and was extracted 3x with 3.5 L aliquots of 1N HCI. The combined aqueous extracts were filtered to give a white solid. The recovered white solid was added back to the organic layer. The 20 L reactor was fitted with a distillation head and the organic layer was distilled to remove in succession: 13.5 L of EtOAc, after which 5 L of heptane was added to the reactor; 5 L of EtOAc/heptane, after which 5 L of heptane was added to the reactor; and 2.7 ' L of EtOAc/heptane, after which 2.7L of heptane was added to the reactor. The contents of the reactor were cooled to 25αC and the resulting mixture was filtered under 5 psig nitrogen while washing with 4 L of heptane. The wet cake was dried under nitrogen pressure overnight to give 1521 g of the titled Compound: 1H NMR (DMSO) 7.12-6.94 (m, 8H), 6.834 (dd, 2H, J = 7.813, 1.709), 6.060 (d, 1H, J - 8.057), 6.050 (td, 1H, J = 7.447, 1.221), 5.795 (d, 1H, J = 8.057), 2119-2.064 (m, 2H), 1.778 (d, 3H, J = 0.997), 1.394 (m, 2H), 0.874 (t, 3H, J = 7.324); Anal. Calod for C22H53N1O3; C, 75.62; H1 6.63;
N, 4.01. Found: C, 75,26; H, 6.72; N, 3.95,
(4S,5R)-3-<2-(E)-Methyl-hex-2-enoyO-4I5-<(ipheny|Oxazolidin-2-^ne (Alternative B)
To a solution of (E)-2-methyl-2-hexenoic acid (6.0 g, 47 rnmol) in 250 mL of THF at 0*C was added 16.3 mL (117 mmol) of triethylamine, then 5.8 mL (47 rnrnol) of pivaloyl chloride resulting in a thick suspension. The mixture was stirred for 1 hour at 00C at which time 2.0 g (47 mmol) of lithium chloride was added in one portion, followed by 10,0 g (42 rnmol) of (4S,5R)-4,5-dϊphenyI-2-oxazαlidinone in four batches. Stirring was maintained throughout the solid additions. The resulting mixture was stirred for 1 hour at 00C1 then for 1 hour at ambient temperature, and was vacuum filtered through a coarse frit and concentrated. The residue was partitioned between EtOAc/water, and the orgaπics were dried over MgSO4 and concentrated, To the residue was added 100 rnL of methyl tert-butyl ether (MTBE) and the mixture warmed cautiously with swirling. The warm slurry was filtered to provide 10.5 g (64% yield) of the titled Compound as a colorless solid: 1H NMR (CDCI3) S 7.12 (rπ, 3H)1 7.07
(m, 3H), 6.94 (m, 2H)1 6.84 (m, 2H), 6.17 (m, 1H), 5,89 (d, J= 7.6 Hz, 1H), 5.68 (d, J = 7.8 Hz, 1H), 2.18 (m, 2H), 1.92 (s, 3H)1 1.50 (m, 2H), 0.96 (t, J = 7.6 Hz, 3H).
(4S,5R)-3-((2R,3R)-2,3-Dimethyl-hexanoyl)4,5-dϊph6nyI-ox;azolidliπ-2-one A 22 L 4-neck round bottom flask was equipped with an addition funnel, mechanical stirrer, and nitrogen inlet. The system was purged with nitrogen for 1 hour. THF (6 L) were charged to the flask followed by 1236 g (6.01 rnol) OfCuBr-S(CHg)2 and 364 g (8,59 mol) of LiCI. The reaction was stirred for 1 S minutes at ambient temperature. The solution was cooled to -35°C and 3.96 L (11.88 mol) of a 3M solution of CH3MgCI in THF was charged at a rate as to keep the internal temperature of the reaction mixture below -25DC. The reaction was stirred for 1 hour after the addition of CH3MgCI was complete. (4S,5R)-3-((E)-2-Methyl- hex-2-er»oylH,5-diphenyI-oxazolidin-2-one (1,00 Kg, 2.86mol) was added as a solid in one portion and the reaction was stirred at -300C for 4 hours. The reaction mixture was transferred over a 2 hour period into another 22 L flask equipped with a mechanical stirrer, transfer line, vacuum line, and containing 4 L of 1:1 acetic acid; THF solution cooled in an ice- water bath. The quenched solution was stirred for 30 minutes and then diluted with 4 L of 2M NH4OH in saturated aqueous NH4CI and 2 L of water. The biphasϊc mixture was stirred for 15 minutes and the phases separated. The organic phase was washed 4x with 4 L aliquots ofthe2M NH4OH solution. No more blue color was observed in the washes or the organic phase so the organic phase was diluted with 8 L of water and the THF was distilled off until the internal temperature of the distillation pot reached 950C. The suspension was cooled to ambient temperature and filtered. The solids were washed with 4 L of water and suction dried to give 868.2 g of an off white solid, This material was recrystallized from 2 L of 95;5 heptanevtoluene with a cooling rate of 5DC per hour to provide 317,25 g of the titled Compound as a white solid: 1H NMR (CDCI3) 7.12-6,85 (m, 10H), 5,90 (d, 1H, J=8,06Hz),
5.72 (d, 1H1 J=7.81), 3.83-3.76 (m, 1H), 1.95-1.89 (m, 1H)1 1.35-1.31 (m, 1H). 1.11 (d, 3H, J=6.84), 1.10-0.95 (m, 3H), 0.92 (d, 3H1 J=6.59), 0.76 (t, 3H, J-7.20) MS (APCl) M+1=366.2.
(2R,3R)-2,3-Dimethyl-hexan-1 -ol A 1 M THF solution of LAH (95,9 ml_ 95,9 mmol) was added to (R)-3-((2R,3R)-2,3- dimethyl-hexanoyl)-4-phenyl-oxazo!idin-2-one in THF (30OmL) under nitrogen at -78°C and stirred for 3 hours at that temperature. Water was added drop-wise to quench the excess LAH and the solution was poured into a mixture of ice and ether. The mixture was extracted into ether, washed with water and dried over MgSO*. The solution was concentrated followed by the addition of excess hexane. The resulting white precipitate was filtered and washed with haxane. The filtrate was concentrated to afford the titled Compound (5.05 g, 100% yield) as an oil: 1H NMR (400 MHz, CDCI3) δ ppm 0.9 (m, 9 H) 1,0 (d, J=6.8 Hz, 1 H) 1,1 (m, 1 H) 1.2 (m, 3 H) 1.6 (m, 2 H) 3.4 (m, 1 H) 3.6 (m, 1 H).
(2R,3R)-2,3-Dime.hyl-hexaπa!
Pyridiπium chlorαchromate (27.35 g, 126.S mmol) and neutral alumina (96 g, 3,5 g per gram of pyridinium chlorochromate) in dry dichloromethane (200 mL) was stirred under nitrogen for 0.25 hours. (aR^R^.S-Dimethyl-hexan-i-ol (5.0 g, 38.46 mmol) in dichloromethane (60 mL) was added and the resulting dark slurry was stirred at room temperature for 3 hours. The slurry was filtered through a short pad of silica eluting with excess dichloromethane. Evaporation of the solvent afforded the titled Compound (4.1 g, 64% yield) as an oil: 1H NWlR (400 MHz1 CDCI3) 5 ppm 0.8 (m, 3 H) 0.9 (d, J=6.6 Hz, 3 H) 1.0 (d, J-6.6 Hz1 3 H) 1.2 (m, 4 H) 1.8 (m, 1 H) 2.2 (m, 1 H) 9.6 (S, 1H).
(S)-N-(1-JPheny!ethyI)-hydroxy|amiπe oxalate [Uskokovic, Tetrahedron 1985, 41, 3455]
A mixture of 20 g (166 mmol) anhydrous magnesium sulfate, 60 mL dichloromethane, 9.63 g (80 mmol) (S)-alρha-meihylbenzylamine, and 11 g (80 mmol) p-aπtealdehyde was stirred under nitrogen at room temperature overnight. The mixture was filtered through a pad of anhydrous MgSO1, washing with 140 mL of dichloromethane. The filtrate was cooled to 0αC under nitrogen and 20,8 g (121 mmol) of 85% meta-chloroperbeπzoio acid slurried in 40 mL dichloromethane was added (slight exotherm). Stirring was continued for 1.5 h at which time the cooling bath was removed and stirring continued for 2.5 hours. The mixture was filtered and the solid washed with 50 mL dichloromethane. The filtrate was washed with 100 mL 0,5 M sodium sulfite, 100 mL 0,5 IW potassium carbonate, 100 mL brine, dried over anhydrous sodium sulfate, filtered and the solvent removed by rotary evaporation (bath temp <30DC). The residue was dissolved in 100 ml absolute ethanol, then cooled to 0gC under nitrogen and treated with 7.55 g (109 mmol) hydroxylamine hydrochloride. The mixture was stirred overnight at ambient temperature. Dichloromethane (150 mL) was added to precipitate excess hydroxylamine hydrochloride and allowed to sit 2 hours then filtered and concentrated.
The resulting oil was taken up in 50 ml water and washed with 2x50 mL diethyl ether. The aqueous layer was poured into 50 mL saturated sodium bicarbonate to give a solution of pH 7-8 (add more sodium bicarbonate if necessary) to give a white solid which was extracted with 4x50 mL diethyl ether. The ether layers were combined and washed with bπne, dried over anhydrous sodium sulfate, filtered into a flask containing 9.4 g (104 mmol) oxalic acid dissolved in 60 mL diethyl ether. The resulting white solid was collected by filtration and dried in vacuo at 500C to give 12.1 g (67%) of the title Compound. MS (ion mode: APCI) m£=i79 [M+H], 1H-NMR (CD3OD) 7.5-7.4 (m, 5H), 4.51-4,46 (q, 1H, J = 4.2), 1,66-1.64 (d, 3H1 J = 4.2), [(2R, 3R)-DimethyIhexyHdine]-2-(S)-methyIbeπ-;ylamine N-oxide [Overton, J. Chem. SOB, Perkin Trans. 7 1991, 1041]
(2R,3R)-2,3-dimethylhexanal (3.33 g, 26.4 mmol), 6.00 g (26.4 mrnol) (S)-N-(I- Phenylethyl)-hydroxylamine oxalate and 4.0 mL (29 mmol) triethylamine in 40 mL dichloromethane was stirred at room temperature overnight. The reaction solution was washed with 25 mL saturated sodium bicarbonate solution and 25 mL brine, dried over anhydrous sodium sulfate, filtered, and chromatographed with ethy! acetate in hexaπe. Appropriate fractions were combined and concentrated to an oil, 5.50 g (85%). MS (ion mode; APC!) mte* 248 [M+H]. 1H-NMR (CDCI3) 7.43-7.24 (m, 5H), 6.60-6.55 (m, 1H), 5.00-
4.93 (rri, 1H), 3.09-3.02 (m, 1H), 1.79-1.76 (m, 3H)1 1.63-1.57 (m, 1H)1 1.37-0.77 ( m, 11H).
3-(S)-[I-(R), 2-(R)-Df rnethylpentyl]-2-(S)-1 -phenylethylj-ϊsoxazolϊdiπ-5one
[2-(R), 3-(R)-Dimethylhexylidine]-2-(S)-methylbeπzylamiπe N-Oxide (4,30 g, 17.4 mmol) and 63 mg (0.17 rnmol) zinc triflate in 50 mL dϊchlorometharie at O0C were treated with
7.6 mL (35 mmol) 1-{ferf-buty|dimethylsϊlyloxy)-1-rnethoxy-etheπe, stirred for 5 rnin and then the cooling bath was removed. Stirring was continued at ambient temperature overnight. The solution was washed with water and brine, concentrated to an oil, and then dissolved in 45 mL THF. 2N aqueous hydrochloric acid (45 mL) was added and the mixture heated at 600C for 2 hours. The cooled mixture was diluted with 50 mL ethyl acetate, the layers separated and the organic layer washed with water and brine, dried over anhydrous sodium sulfate, filtered, concentrated, and chromatographed on silica gel with ethyl acetate/hexane to give an oil which was crystallized from 20 ml hot 80% methaπol/waterto give 2.26 g (45%) of a white solid. MS (ion mode; APCi) mfz~ 290 [M+H]. 1H-NMR (CDCI3) 7.35-7.24 (rn, 5H), 4.03-3.98 (q, 1H, J = 4.2), 3.38-3.32 (m, 1H), 2.40-2.34 (dd, 1H, J = 4.5 and 11.1), 2.08-2.02 (dd, 1H, J
= 5.2 and 11.1), 1.63-0.77 (m, 1SH).
(3S,4R,5R)-3-Am(πo-4s5-dirnethyloctaπoic acid
3-(SHI-(R), 2-(R)-DimethylpentylJ-2-(S)-1-phenylethyl]-isoxaZθlidiπ-5-orιe (2.26 g, 7.8 mmol) was hydrogeπated for 17.8 h with 0.8 g 20% palladium on carbon in 100 mL of ethanol. The mixture was filtered, concentrated to a white solid which was crystallized from 10-12 mL hot methanol and acetonitrile (8:2) to give 1.26 g (86%) of a white solid. MS (ion mode: APCI) m/z= 138 [M+H]. 1H-NMR (CD3OD) 3.47-3.42 (m, 1 H), 2.45-2.40 (dd, 1H, J = ZO and 10.5), 2.24-2.20 (dd, 1H1 J - 6.5 and 10.5), 1.63-1.02 (m, 6H), 0.96-0.88 (m, 12H). The following illustrations of the invention are provided by way of example and not by way of limitation. EXAMFLES
The following methods, results, and summary are presented as examples of in vitro receptor binding and behavioral effects of the Ot2S-I selective ligand, Compound A, compared to the α2δ non-selective ligand, (SS.δRJ-S-aminomethyl-e-cyclopropyl-δ-methyl-hexaπαie acid, hereinafter "Compound B", as described in WOQO/076958.
Figure imgf000021_0001
Compound B
ME=THODS Receptor Binding:
HEK 293 cells stably expressing recombinant porcine α2δ-1 and human α2δ-2 subuπϊts were constructed previously (Gong KC. etal. (2001) J. Membr. Biol. 184:35-43) were grown under normal cell culture conditions (RPMI-1640 media with io% FBS, 200μg G418, and 1% penicillin/ streptomycin at 37° C with 5% CO2) until reaching 90% confluency in T-75 flasks, at which time they were harvested. Cells were suspended in ice-cold 5mM
Tris/5mM EDTA buffer, pH 7.4 (TE buffer) containing PMSF (0.ImM) and protease inhibitor cocktail (Roche) and allowed to sit on ice for 30 minutes. Cells were broken by sonication using 20 bursts, 40-50 cycles, and then centrifuged at 3000 x g for 10 min. The supernatant was transferred to a newtube and centrifuged at 50,000 x g for 30 min. The resulting pellet was re-suspended in 1OmM HEPES buffer, pH 7,4 and homogenized before storage at-
800C. Protein concentration was measured by well known methods.
The fH]-gabapentin SPA binding assay was performed in Costar 363296-well, clear bottom assay plates using wheat germ agglutinin coated polyvinyl toluene scintillation proximity assay (SPA) beads (Amersham Biosciences). Alpha-2-delta-1 or o^δ-2 membrane proteins (10-20 μg protein per well) and SPA beads (0,5 mg per well) were mixed with 30 nM
[*Hl-gabapentin (60 Ci/mmol; synthesized by Pfizer Global Research and Development Michigan Labs) in 10 mM HEPES/10mM MgSO4 assay buffer, pH 7.4 using KOH. The final well volume was 200 μl and non-specific binding was determined in the presence of 10 μM unlabeled (cold) pregabaliπ. The mixture containing membrane protein with SPA beads and f HJ-gabapentiπ was incubated at room temperature over night (16-20 hrs), and plates were then counted on a Wallace Trilux 1450 Microbets scintillation counter. Curve fitting and IC50 values were calculated using a four-parameter, non-linear regression equation from GraphPad Prism 4.0 software, while K-, values were determined using previously
ra Λ ΛO determined K0 values for α2δ-1 (K0 = 41 nM), and α2δ-2 (KD = 146 πM) and the equation of Cheng and Prussoff (Bioehem Pharmacol. (1973) 22(23): 3099-3108.
Behavioral Testing Animals:
Male, C57BL/6J mice (Jackson Laboratories, Bar Harbor, Maine) were used for vogel conflict, locomotor activity and accelerating rotarod testing. Animals were received at 5 weeks old and acclimated to the facility 1 week prior to testing. Male, DBA/2J mice 3 week old (Jackson Laboratories, Bar Harbor, Maine) were used for evaluating anticonvulsant activity, Mice were housed 5/isolator in a temperature/humidity-controlled room under a 12; 12 hour lighfcdark schedule (lights op at 6:00 AM) with food and water available ad libitum. AH procedures were carried out in compliance with the NlH Guide for the Care and Use of Laboratory Animals under a protocol approved by the PGRD Animal Use Committee.
Vogel Apparatus:
The test apparatus consists of 12 modular operant chambers (Coulboum Instruments). The front and back of the test chambers are made of clear Plexiglas. The front doors are covered to reduce distractions from inside the test room. The backs face a wail, away from the flow of traffic in the testing room and remain uncovered to provide the opportunity for observations. All chambers have stainless steel grid floors and measure 7 x 7 x 12 inches. Each test chamber is modified with an internal chamber made of clear Plexiglas measuring 6.75 x 3.5 x 1.5 inches. The reduced chamber space limits the animal's activity and directs behavior towards the opening on the side of the chamber 1.5 inches above the floor. A module optical lickometer mounted at the opening is used to measure licking. A water bottle attaches to the outside of the cage and the drink tube extends into the opening through the module. The reinforcer consists of a 1:1 mixture of evaporated milicwater. A photo beam is piped across two glass rods that reside adjacent to the tip of the drink tube. Each time the animal licks the drink tube the beam is broken and licks are automatically recorded. Shock is delivered between the grid floor and the drink tube using a (Coulboum Instruments) programmable universal shocker for one second, but terminates Immediately when contact between the animal and drink tube is broken.
Vogel Procedure: On day one, after 24 hours of water deprivation, experimental subjects were placed in the test chambers and allowed to drink unpunished for a 10-minute training session. Mice were required to complete a 100-150 lick criteria during the training session. Subjects that completed the training criteria prior to the end of the 10-rninute session were removed from the test chamber. This served to restrict intake in all subjects to -25% Qf their total daily coπsumption. Mice that failed to complete 100 licks or exceeded 150 licks were eliminated from the study. Immediately after the day one session mice were returned to their home cages and water deprivation continued. All mice were then fasted overnight prior to testing. On test day two, mice were dosed with vehicle or test Compound and tested 30 minutes later, During the 10-mϊnute test session mice received a mild shock (0.4 mA) every 10m lick [fixed ratio (FR) 10 schedule]. Hence, a conflict situation exists; mice were motivated to drink but drinking was inhibited by the shock. The low number of shock episodes received reflects anxiety-related behavior. Standard anxiolytic drugs significantly increase the number of shock episodes over the concurrent vehicle control group and produce an aπxiolytic-Iϊke effect Group means + SEWI (N=12/group) were calculated and all data were analyzed using Anova on Ranks/Dunn's Method compared to the concurrent vehicle control group.
Anticonvulsant Activity (DBA/2J Mice)
Immediately before anticonvulsant testing, mice were placed upon a wire mesh, 4- inch square, suspended from a steel rod 12 to 18 inches from the top of a table. The square was slowly Inverted through 180 degrees and mice observed for 30 seconds. Any mouse falling from the wire mesh was scored ataxic.
Anticonvulsant testing started by placing individual mice into an enclosed acrylic plastic chamber (21-cm height, approximately 30-crn diameter) with a high-frequency speaker (4-cm diameter) in the center of the top lid, An audio signai generator (Protek Model B-810) was used to produce a continuous sinusoidal tone that was swept linearly in frequency between 8 and 16 kHz once each 10 milliseconds. The average sound pressure level duing stimulation was approximately 100 dB at the floor of the chamber. Mice were placed within the chamber and allowed to acclimatize for 1 minute. DBA/2J mice in the vehicle-treated group responded to the sound stimulus (applied until tonic extension occurred, or for a maximum of 60 seconds) with a characteristic seizure sequence consisting of wild running followed by clonic seizures, and later by tonic extension, and finally by respiratory arrest and death in 90% or more of the mice. In vehicle-treated mice the entire sequence of seizures to respiratory arrest lasted approximately 10 to 15 seconds.
Locomotor Activity:
Locomotor activity (LMA) testing is performed using 16-Beam Digiscan Animal Activity Monitors (Acouscaπ Electronic, Colombus, OH). Each chamber consists of a Plexiglas box measuring 16 x 16 inches with a Plexiglas insert that divides the box into 4 equal quadrants 7.5 x 7,5 inches each. Two mice are placed in diagonal quadrants for testing with the lights on and infrared beams located on the perimeter of the chamber detect movement The entire test chamber is enclosed in a sound reduction chamber to reduce extraneous noise during the test session. Prior to the test day all mice are fasted overnight On the test day mice receive vehicle or test Compound 30 minutes prior to testing and total distance (cm) is recorded in 5-minute blocks for 1 hour, Data is reported as group Means + SEM
(N=10/group) and subjected to One Way Anova/Duππett" s or Anova on Ranks/Dunn's versus the concurrent vehicle group for statistical analysis as appropriate.
Accelerating Rotarod:
The test equipment consists of four programmable SmartRod® chambers (AccuScan Instruments, Columbus, OH). Each chamber is 36 cm (h) x 11 cm (w) x 30 (d) and equipped with a rotating rod horizontally affixed 32,5 cm on center above the grid floor. The rod spans the 11 cm width of the chamber and is 3 cm in diameter. The equipment is programmed to accelerate the rod at a rate of 0.25 rev/sec to a maximum speed of 20 rpm over 62 seconds, after which the rod decelerates over the final 5 seconds to the stop position. The total cycle time for one trial is 67 seconds. For each trial the mouse is suspended by the tail directly above the rod for three seconds simultaneous to the start of the cycle, then gently lowered onto the rotating rod. Mice must traverse the rod or fall to bottom of the test chamber. Two centimeters above the floor of the test chamber reside two infrared (IR) beams linked to a programmable timer. A mouse that falls from the rod breaks the IR beams and triggers the automatic shut off of the timerto record fall latency (seconds), One day prior to testing mice must complete 2 of 6 consecutive training trials with fall latency greater than 10 seconds. Animals that complete the training criteria are fasted overnight Day two testing consists of the mean of 3 consecutive trials on the rod following a 30-miπute pretreatmeπttime. Data are reported as group Means, + SEM (N=12/group) and subjected to One Way Anova/Dunnett's or Anova on Ranks/Dunn's versus the concurrent vehicle group for statistical analysis as appropriate,
Compounds:
Compound A and Compound B were each separately dissolved in sterile saline and each dose was delivered in a total volume of 5 μl_ at doses of 3 -30 μg/mouse. Vehicle control mice received 5 μl_ of sterile saline. A 27g steel needle calibrated to a depth of 3 mm was attached to a 25 μL Hamilton syringe for intraeerebroventricular (ICV) administration. Mice were manually restrained and the injection site was found by locating bregma on the skullcap with the tip of the needle. One mm to the lateral and 2 mm to the posterior of bregma the needle was dropped through the skull and into the ventricles and drug or vehicie was administered in 5 μL bolus.
RESULTS
Receptor Binding
Compound A was found to displace [3H]-gabapentiπ binding with moderate-to-high affinity to membrane proteins prepared from recombinant porcine α2δ-1 cells, whereas Compound A displaced [3H]-gabapentin binding at very low affinity to membrane proteins prepared from recombinant human α2δ-2 cells. The K, values for displacement of binding of [3H]-gabapentiπ to recombinant porcine Ot2S-I and recombinant human <*&-1 cell membranes were 35 nWl, and 1670 πM, respectively, Selectivity for the α2δ-1 is 48-fold greater than for the α2δ-2 site, in comparison, the non-selective Compound (Compound B) displaced [3H]- gabapentin in each of the membrane preparations with a Ki value of 32.8 nM for recombinant porcine α2δ-1 and 34.9 nM for recombinant human αzδ-2 cell membranes. The Kj value determined for gabapentin was 75 nM for recombinant porcine aφ-Λ, and 114 nM for recombinant human α2δ-2 cell membranes (all values in this section are mean),
Behavioral Testing
Compound A was compared to Compound B in the Vogel Conflict Test, sound- induced tonic seizures, and the locomotor activity and rotarod adverse effect models. The effects of ICV dosing on C57BL/SJ mice following a 30-minute pre-treatmeπt time using Compound A and Compound B in the Vogel Conflict test are shown in Tables 1 and 2, respectively.
Figure imgf000025_0001
Figure imgf000025_0002
The α2δ-1 selective ligand, Compound A (330 μg/mouse), produced a dose dependent aπxiolytic-like effect in Vogel Conflict as reflected by an increase in mean shock episodes compared to the concurrent vehicle control group (Table 1). The vehicle control group mean + SEWl was 10.8 + 1.1. Statistical significance was observed at 10 and 30 μg/rnouse (Mean ± SEM = 33.3 + 5.3 and 43,4 + 3.0, respectively).
The non-selective Compound, Compound B {0.33 μg/mouse), produced a dose dependent anxiolytiσ-like effect in the Vogel Conflict model reflected by an increase in mean shock episodes compared to the concurrent vehicle control group (Table 2), The vehicle control group mean ± SEM was 9.9 + 1.0. The minimally effective dose (MED) for Compound B was 3 μg/mouse with a magnitude of the response at this dose of 31,2 + 8.2. Therefore, both compounds were shown to produce an anxiolytiotike effect.
The effects of ICV dosing on DBA/2J mice following a 2-hour pre-treatmeπt time using Compound A and Compound B on the prevention of sound-induced seizure tonic seizures is shown in Tables 3 and 4, respectively.
Figure imgf000026_0001
Figure imgf000026_0002
Compound A (3-30 μg/rnouse), produced a dose dependent increase in protecting DBA/2J mice from tonic seizures. The ED50 value for Compound A was determined to be 6.6 μg ICV; with a S5% confidence interval of [4.2 to 10.3]. Similarly, Compound B (3-30 μg/mouse) produced a dose dependent increase in protection of DBA/2J mice from tonic seizures. The ED50 value for Compound B was determined to be 11.3 μg ICV; with a 95% confidence interval of [5.7 to 2ZT].
The affects of ICV dosing on C57BU6J mice following a 30-minute pre-treatmerrt time using Compound A and Compound B on locomotor activity and the accelerating rotarod activity test is shown in Tables 5 and 6, and 7 and 3, respectively.
Table 5. Effects of Compound A on locomotor activity (adverse effect)
Treatment Group Dose (μg/mαuse)a Route Mean Distance Traveled (cm) D
Vehicle - ICV 2,386.9 + 273.5
Compound A 3 ICV 2,634.7 + 266.8
Compound A 10 ICV 2,214.0 + 179.2
Compound A 30 ICV 2,444.3 + 318.7 aDosed in 5 μL volume bData are mean ± SEM N=10/group *= pθ.05 ANOVA/post hoc versus concurrent Vehicle
Figure imgf000027_0001
Overall, there was an 11.4 to 84.3% reduction in mean distance traveled in mice administered escalating doses (3 - 30 μg /mouse, ICV) of Compound B1 relative to the vehicle. Statistical significance was observed at doses of 10 and 30 μg /mouse. The MED for Compound B was 10 μg/mouse. Compound A1 (3 - 30 μg /mouse, ICV) had no effect on mean distance traveled across the entire dose range tested. Therefore, the locomotor adverse effect was avoided in mice administered an αsδ-1 selective ligand. Higher doses were not explored due to lack of solubility at higher concentrations. Table 7. Effects of Compound A in the accelerating rotarod test (adverse effect)
Treatment Group Dose (μg/rnouse)a Routθ Mean Fall Latency (sec) "
Vehicle — ICV 54.5 + 4.0
Compound A 3 ICV 41.1 + 4.2
Compound A 10 ICV 49,0 + 2.6
Compound A 30 ICV 42.9 + 4.2
"Dosed in 5 μl_ volume "Data are mean ± SEM N=i2/group *= p<0.05 ANOVA/post hoc versus concurrent Vehicle
Table 8. Effects of Compound B in the accelerating rotarod test (adverse effect)
Treatment Group Dose (μg/mouse)a Route Mean Fall Latency (sec)"
Vehicle — ICV 43.7 + 4.2
Compound B 3 ICV 33.0 + 3.0
Compound B 10 ICV 31.6 + 3.3*
Compound B 30 ICV 20.2 + 1.8* aDosed in 10 μL volume "Data are mean ± SEM N=12/group •a p<0.05 ANOVA/post hoc versus concurrent Vehicle
Overall, there was a dose dependent decrease in mean fall latency in mice administered escalating doses (3-30 μg/mouse, ICV) of Compound B, relative to the vehicle.
Statistical significance was observed at doses of 10 and 30 μg/mouse, which resulted in a 24.5 and 53.3% decrease in mean fall latency compared to vehicle. The MED for Compound B was 10 μg/mouse. Compound A had no statistically significant effect on mean fell latency across the entire dose range. Therefore, administration of an Ci2S-I selective ligand was shown to have an improved adverse effect profile. Overall, the frequency or incidence
(percent) of one or more adverse effects following administration of an O2S-I selective ligand may be reduced by at least 10 percent or greater.
Compound A is an α2δ-1 selective ligand. In competition binding studies to displace [3H]-gabapentin, Compound A prefers recombinant porcine O2S-I protein with binding inhibition Ki of 35 nM, whereas its binding inhibition Ki to recombinant human 036-2 protein was 48-fold less at 1670 nM. In contrast, the non-selective α2§ ligand like Compound B showed similar binding affinity to both the Ot2B-I (K, = 32.8 nM) and α2δ-2 (K,- = 34.9 nM). In behavioral testing, both Compound A and Compound B produced robust anxiolytic-like effects in C57BL/6J mice in the Vogel Conflict model and anticonvulsant activity in the mouse DBA/2 sound-induced seizure model following ICV administration. In contrast, at doses required for anxiolytic and anticonvulsant activity, Compound B had significant activity in two behavioral models commonly used to evaluate potential adverse effects, accelerating rotarod and locomotor activity, while Compound A showed no significant effect in these adverse effect rnodels. In conclusion, these data show an improved therapeutic window for Compound A, a compound with preference for α2δ-1 protein; by exhibiting robust arrxiolytϊc-like efficacy with reduced locomotor and motor control adverse effects when compared to an o.2δ non-selective ligand.
While the forms of the invention herein disclosed constitute presently preferred embodiments, many others are possible. It is not intended herein to mention all of the possible equivalent forms or ramifications of the invention. It is understood that the terms used herein are merely descriptive, rather than limiting, and that various changes may be made without departing from the spirit or scope of the invention. All numerical ranges described herein having one or more endpoϊnts include the endpoiπts and all numerical values between the endpoints. Ail literature citations cited herein are hereby incorporated by reference.

Claims

What is claimed is:
1, A method of treating a disorder or condition in a mammal, including a human, comprising administering to said mammal a therapeutically effective amount of an alpha-2- delta-1 selective ligand, or a pharmaceutically acceptable salt thereof; wherein said disorder is selected from pain, fibromyalgia, epilepsy, restless legs syndrome, hot flashes, mood disorders and sleep disorders.
2, The method according to claim 1, wherein the alpha-2-delta~1 selective ligand is
(3S,4R,5R)-3-Amino-4,5-dϊmethyloctaπoic acid or a pharmaceutically acceptable salt thereof.
3. The method according to claim 1, wherein said a!pha-2-delta-1 selective ligand is at least 30-, 40-, 5Q-, 100-, 50CK or 1000-fold more selective for the a(pha-2-delta-1 subuπlt than the alpha-2-delta-2 subunϊt.
4. The method according to claim 1, wherein said alpha-2-delta-1 selective ligand binding is at least 48-fold more selective for the alpha-2-defta-1 subuπit than the alphas- delta^ εubuπit,
5. The method according to claim 1 wherein said pain disorder is neuropathic pain,
6. The method according to claim 1 wherein said sleep disorder is insomnia.
7 The method according to claim 1 wherein said disorder is hot flashes
8 The method according to claim 7, wherein hot flashes is surgically or drug induced.
9 The method according to claim 1 wherein said mood disorder is selected from generalized anxiety disorder and social anxiety disorder.
10 The method according to claim 1, wherein said disorder is epilepsy.
11 The method according to claim 1 wherein said disorder is restless legs syndrome.
12 The method according to claim 1 wherein said disorder is fibromyalgia.
13 The method according to claim 1 wherein said alpha-2-delta-1 selective ligand is administered in an amount of 10 to 5000 mg per day. 14 The method according to claim 1, wherein said administration is carried out orally, parenterally, subcutaneously, intravenously, intramuscularly, intraperitoneal^, by intranasal instillation, by implantation, by intracavitary or intravesical instillation, intraocularly, intraarterially, intralesioπally, transdermally, or by. application to mucous membranes.
15 The method according to claim 1, wherein said alρha-2-delta-1 selective ligand is present in a pharmaceutical composition comprising the ligand and a pharmaeeutically- acceptable carrier.
16 The method according to claim 15, wherein the pharmaceutical composition is in a liquid or solid dosage form.
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