JP2005320345A - Phosphodiesterase iv inhibitor for treating multiple sclerosis - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a new pharmaceutical composition for treating multiple sclerosis. <P>SOLUTION: The pharmaceutical composition comprises an effective amount of a PDE IV inhibitor, an anti-inflammatory or immunomodulatory drug and a pharmaceutically acceptable carrier. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to PDE IV inhibitors for the treatment of multiple sclerosis.

  Demyelination is a serious disease of the brain and spinal cord that involves breakage of the myelin sheath that surrounds nerve fibers. As a result of demyelination, various neurological symptoms appear, including movement disorders, visual loss and sensory changes. Multiple sclerosis (MS) is the most common form of demyelination. It is a disease characterized by optic nerve focus disorder, spinal cord and brain pathogenesis. This is a severe chronic impotence with a characteristic demyelination in the CNS caused by an autoimmune mechanism that can occur in a genetically ill population. Typically, this results in remission (relapse-reduction) recurrent pathogenesis following neurological dysfunction, but it can also be chronic. Although MS is a well-studied disease, the detailed course remains unclear. One explanation is that the most frequent environmental factors in childhood are specific populations of T-cells that are normally controlled by suppressor cells (myelin-associated antigens such as MBP, MAP, MOP or other attack potentials). Is activated). In many MS cases, the result is that nonspecific stress results in BBB initiation, edema, disease progression with T-cell and macrophage transfer and activation, and subsequent destruction of oligodendrocyte-associated myelin Attempts to regenerate myelin are unsuccessful, with a final glial scar (MS recurrence-reduction). These disease progressions that can be visualized by MRI are associated with significant functional failure and increased recurrence depending on the location, and this disease (and the antigen being attacked) is more generalized and has a smaller clear cut interval ( Progressing in MS progressive form (secondary or primary) and increasing and persisting irreparable, causing life damage (in almost young populations), loss of employment and independent life (hospital life and cases) With death).

  Until recently, treatment has been done experimentally but with no success. See Cecil's Textbook of Medicine (Wyngaarden, 1993). Classic non-specific immunosuppressive therapies (including corticosteroids) and static cell drugs usually fail to alter this set of conditions and disease course, probably because they are This is because it is not well tolerated by patients and similarly suppresses endogenous immune suppression mechanisms as well. See, for example, Principles of Neurology, 5th Edition (Adams et al., 1993) for treatment considerations. Recent non-specific immunodegenerative treatment with interferon-β-1b has been shown to prolong disease-free periods in patients with early relapse / relief MS, but in the majority of these patients, The worsening of the lesion cannot be completely prevented, and the effect on the inability of the first three years of treatment is still small. Corticosteroid treatment alleviates some acute symptoms of these disease progressions (possibly due to reduced edema) but does not work for long-term prognosis. However, it seems most important to suppress all disease progressions that are accompanied by activation, extension and augmentation of the autoimmune mechanisms that render this disease progression uncontrollable and impossible. There also remains a need for additional drugs that affect the severity and progression of this disease.

Recent advances in clinical, biochemical and imaging techniques have allowed clinicians to predict and diagnose such disease progression at a very early stage, and thus various approaches to achieve maximum therapeutic effect. It became possible to combine treatment plans. Immunodegenerative treatment using non-specific mechanisms using, for example, interferon-β-1b (or a specific specific immunodegenerative agent such as Copolymer I, as well as inducing tolerance to MS antigens) is Although not acting up to 100% as needed before, the number of clinical disease progression can be clearly and significantly reduced even in lesions in the brain. Fortunately, with regard to targets for fully understanding disease activation, new diagnostic techniques that have been developed (e.g., increased γ-interferon, TNF-α, activated specific T-cells in the blood) Increased numbers, new specific MRI and other imaging techniques and clinical observations) have the ability to detect the onset of disease progression very early.
Cecil's Textbook of Medicine (Wyngaarden, 1993) Principles of Neurology, 5th Edition (Adams et al., 1993)

  Therefore, there is a need for powerful and effective drugs that can not only act on some symptoms of disease progression, but completely block them and thus control disease progression. However, in autoimmune conditions, such as peripheral T-cell activation, endothelial adhesion, blood-brain barrier opening and T-cell-macrophage / microglia interaction activation, subsequent oligodendrocyte injury and demyelination New treatment regimes that are intended to act on protective mechanisms in chronic use, but not against endogenous extrinsic drugs and effects (eg tumorigenesis) as well as damage to extrinsic (eg bacterial, parasite or viral infection) It is also a serious defense mechanism.

  One aspect of the present invention combines well-tolerated long-lasting treatment with the use of novel diagnostic methods to predict or detect early disease progression that can only be aggressively treated with many drugs or combinations thereof. To ensure that it does not produce stubborn CNS lesions that have fatal effects on disease progression, prolongation and progression. Furthermore, the combination of treatment with various mechanisms to achieve maximal effect is more tolerable of treatment (because the effect of interferon-β-1b in MS has been shown to be dose dependent, Higher doses can be expected to achieve a high and 100% effect, but significant increases in side effects prevent this type of treatment), and ultimately this dose maintenance and anti-morbidity treatment The new combination of results in a marked reduction in the risk of side effects that can be caused by high doses and prolonged use of these drugs during monotherapy. In fact, supplemental and synergistic therapies when basic dose maintenance treatment is combined with the use of this new treatment plan only when it is necessary (eg during or just before disease progression) Both side effects rarely occur and this form of treatment can be combined to achieve optimal clinical outcome. In this new concept of short-term treatment of disease progression, various drugs can be combined to prevent further damage and progression. Thus, inhibition of the synthesis of cytotoxic cytokines and chemokines suppresses their release, both of which suppress drugs or eventual tissue damage that can be enhanced by simultaneous inhibition of flow across, for example, the blood-brain barrier- Or it can be combined with a retrograde agent (eg nerve growth factor, calcipotriols, calpain inhibitors, etc.).

  Although many compounds can attack one or more of these MS mechanisms and usually have the typical risks and side effects associated with special mechanisms, our experimental studies have surprisingly shown that inhibition of phosphodiesterase IV Agents such as rolipram show very high activity on human MS cells as well as in various animal models of the disease (eg EAE, EAN in different species); Safety and efficacy on various biological parameters have already been observed. Using these combined mechanisms, low doses, either alone or in combination with other dose maintenance treatments, have very low side effects (others) for the prevention or reduction or treatment of MS disease progression. It is clear that it can be used in comparison to treatment).

  In addition, at the same or lower (exceptionally higher) doses, these drugs have additional and synergistic effects (and thus inhibition of progression as described above) with minimal side effects to achieve maximum effect. Can be combined with other compounds as described in 1-9.

  Accordingly, one aspect of the present invention relates to a method of treating or preventing MS comprising applying an effective amount of a type IV phosphodiesterase inhibitor (PDE IV inhibitor) and an anti-inflammatory or immunodegenerative agent.

Phosphodiesterase PDE inhibitors suitable for use in the present invention are cycloadenosine-- according to the latest classification (JA Beavo and DA Reifsnyder, Trends Pharmacol. Sci. 11; 150-155, 1990). Advantageously, it is a 3 ′, 5′-monophosphate (cAMP) PDE type IV (PED IV) inhibitor; S. 4193629, WO 92/02220, U.S. Pat. S. 4186129; EP247725; S. 5064854; A. Saccamono et al. , J .; Med. Chem. 34, 291-298, 1991; J. et al. Vinick et al. , J .; Med. Chem. 34, 86-89, 1991; A. Lowe et al. , J .; Med. Chem. 34. , 624-628, 1991, including but not limited to:
1,3-dibutyl-3,7-dihydro-7- (2-oxopropyl) -1H-purine-2,6-dione (denbufylline, BRL30892);
4-[(3-butoxy-4-methoxyphenyl) methyl] -2-imidazolidinone (RO20-1724);
5,6-diethoxybenzo [b] thiophene-2-carboxylic acid (tibenlast, LY186655);
3-ethyl-1- (3-nitrophenyl) -2,4 (1H, 3H) -quinazolinedione (nitrazone, TVX 2706);
6- (3,6-dihydro-6-methyl-2-oxo-2H-1,3,4-thiadiazin-5-yl) -1- (3,4-dimethoxybenzoyl) -1,2,3,4 -Tetrahydro-4,4-dimethylquinoline (EMD 54622);
1-ethyl-4-[(1-methylethylidene) hydrazino] -1H-pyrazolo [3,4-b] pyridine-5-carboxylic acid ethyl ester (etazolate);
N-hydroxy-5,6-dimethoxy-benzo [b] thiophene-2-carboxyimidamide (Org 30029);
2-Amino-6-methyl-4-propyl (1,2,4) triazolo [1,5-a] pyrimidin-5 (4H) -one (ICI 63197);
6- [4- (Difluoromethoxy) -3-methoxyphenyl] -3 (2H) -pyridazinone pentoxifylline; propentofylline; vinpocetine; and their pharmacology Tolerable salt.

  Preferred PDE IV inhibitors are those of formula I:

［式中、Ｒ^１は、Ｃ_１〜Ｃ_６−アルキル、ヘテロ環、又はＯＲ^５であり、ここで、Ｒ^５は、Ｃ_１〜Ｃ_６−アルキル、Ｃ_３〜Ｃ_７−シクロアルキル、Ｃ_２〜Ｃ_６−アルケニル、Ｃ_３〜Ｃ_７−アルキニル、Ｃ_３〜Ｃ_７−シクロアルキル−Ｃ_１〜Ｃ_２−アルキル、アリール、アラルキル、ヘテロ環又は１個以上のハロゲン原子で置換されたＣ_１〜Ｃ_６−アルキル、ヒドロキシ、カルボキシ、Ｃ_１〜Ｃ_４−アルコキシ、Ｃ_１〜Ｃ_４−アルコキシカルボニル又はアルキルで置換されていてよいアミノ基であり；
Ｒ^２は、Ｃ_１〜Ｃ_４−アルキル、Ｃ_２〜Ｃ_４−アルケニル又はＣ_２〜Ｃ_４−アルキニルであり；
Ｒ^３は、水素原子、Ｃ_１〜Ｃ_６−アルキル、アリール、アラルキル、又は１個以上のメチル基で置換されていてよいアリール又はＣ_１〜Ｃ_６−アルカノイルであり；
Ｒ^４は、水素原子又はＣ_１〜Ｃ_６−アルキルであり；
Ｙは、直接結合又はＣＨ_２基であり；
Ｘは、ＣＨ_２、ＣＨ_２−ＣＨ_２、ＮＨ又は酸素原子である］のラセミ性及び光学的に活性の化合物及びその薬物学的に認容性の塩である。

[Wherein R ¹ is C ₁ -C ₆ -alkyl, heterocycle, or OR ⁵ , wherein R ⁵ is C ₁ -C ₆ -alkyl, C ₃ -C ₇ -cycloalkyl, C 5 C ₂ -C ₆ -alkenyl, C ₃ -C ₇ -alkynyl, C ₃ -C ₇ -cycloalkyl-C ₁ -C ₂ -alkyl, aryl, aralkyl, heterocycle or C substituted with one or more halogen atoms ₁ -C ₆ - alkyl, hydroxy, _carboxy, C 1 -C ₄ - _alkoxy, C 1 -C ₄ - There alkoxycarbonyl or alkyl substituted may have an amino group;
R ² is C ₁ -C ₄ -alkyl, C ₂ -C ₄ -alkenyl or C ₂ -C ₄ -alkynyl;
R ³ is a hydrogen atom, C ₁ -C ₆ -alkyl, aryl, aralkyl, or aryl optionally substituted with one or more methyl groups or C ₁ -C ₆ -alkanoyl;
R ⁴ is a hydrogen atom or C ₁ -C ₆ -alkyl;
Y is a direct bond or a CH ₂ group;
X is CH ₂ , CH ₂ —CH ₂ , NH or an oxygen atom], and racemic and optically active compounds and pharmaceutically acceptable salts thereof.

Preferred compounds of the formula I are those in which R ² is methyl, R ³ is a hydrogen atom or C ₁ -C ₆ -alkanoyl, R ¹ is OR ⁵ and R ⁵ is C ₁ -C _{6- alkyl,} C 3 -C ₇ - cycloalkyl or 3- tetrahydrofuranyl, ^{R 4} is hydrogen or _C 1 -C ₄ - alkyl, a compound wherein X is CH ₂ group or oxygen.

Particularly preferred compounds of the formula I are those in which R ³ is hydrogen.

  A particular example is 4- (3- (cyclopentyloxy) -4-methoxyphenyl) -2-pyrrolidinone (rolipram).

  The term “alkyl” as used herein includes straight or branched alkyl groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, s-butyl, isobutyl, t-butyl, pentyl, Includes 2-methyl-butyl, 2,2-dimethylpropyl and hexyl.

  The term “alkenyl” as used herein is meant to include, but is not limited to, vinyl, 1-propenyl, 2-propenyl, 2-propynyl or 3-methyl-2-propenyl.

  As used herein, “cycloalkyl” or “cycloalkylalkyl” is meant to include groups having 3 to 7 carbon atoms, such as cyclopropyl, cyclopropylmethyl, cyclopentyl, or cyclohexyl.

  The term “aryl” or “aralkyl” as used herein means an aromatic ring or a ring system having 6 to 10 carbon atoms, preferably a single ring such as phenyl, benzyl, phenethyl or naphthyl.

  The term “heterocycle” as used herein is a 5-6 membered saturated ring having one oxygen, sulfur or nitrogen atom, such as 2- and 3-tetrahydropyranyl, 2- and 3-tetrahydrofuranyl, Means pyrrolidino, 2- and 3-pyrrolidyl, piperidinino, 2-, 3- and 4-piperidyl and the corresponding N-alkylpyrrolidyl and piperidyl rings, wherein alkyl is of 1 to 4 carbon atoms, However, it is not limited to these. Also included within the scope of the invention are heterocycles having one or more heteroatoms, such as morpholino, piperazino or N-alkyl piperazino.

  The term “halogen” as used herein means all halogens, ie chlorine, fluorine, bromine and iodine.

  The preparation of the compound of formula I can be carried out in the manner described in the above patent specification or in US Pat. Nos. 4,153,713; 4,186,129; and 5,298,628; WO 86/02268; or EP 0247725. it can. Rolipram is 4-[(3-cyclopentyloxy) -4-methoxyphenyl] -2-pyrrolidinone. For example, Merck Index, 11th. See edition, pp 1312-1313. Rolipram and related compounds can be prepared, for example, according to US Pat. No. 4,193,926.

Anti-inflammatory and immunodegenerative agents suitable for use in the present invention include, but are not limited to:
1. Interferon derivatives such as betaserone, β-interferon, β-interferon muteins,
2. Prostan derivatives such as compounds described in PCT / DE93 / 0013, such as iloprost, cicaprost;
3. Glucocorticoids such as cortisol, prednisolone, methylprednisolone, dexamethasone;
4). Immunosuppressants, such as cyclosporin A, FK-506, methoxalene, thalidomide, sulfasalazine, azathioprine, methotrexate;
5). Lipoxygenase inhibitors such as zileuton, MK-886, WY-50295, SC-46562, SC-41661A, BI-L-357;
6). Leukotriene antagonists such as, for example, German Patent (DE) 4009117; German Patent Application P4242390.2; compounds described in WO 9201675; SC-41930; SC-50605; SC-51146; LY255283 (Dk Herron et al., FASEB J Abstr. 4729, 1988); LY 223882 (DM Gapinski et al., J. Med. Chem. 33: 2789-2813, 1990); Morris et al., Tetrahedron Lett. 29: 143-146, 1988, C.I. E. Burgos et al. Tetrahedron Lett. 30: 5081-5084, 1989; M.M. Taylor et al. , Prostaglandins 42: 211-224, 1991, U-75302 and analogs; S. 5019573; and e.g. Kishikawa et al., Adv. Prostagl. Thrombox. Leukotriene Res. 21: 407-410, 1990; Konno et al., Adv. Prostagl. Thrombox. Leukotriene Res. 21: 411-414, 1990; ONO-LB-457 and analogs; S. WF-11605 and analogs described in 4963583; compounds described in WO91118601, WO91118879; WO9118880 and WO9118883;
7). Anti-inflammatory substances such as L. Noronha-Blab. NPC 16570, NPC17923 described in Gastroentrology 102 (Suppl.) A672, 1992; N. Buch et al., Proc. Nat. Acad. Sci. USA 88: 355-359, 1991; Pou et al., Biochem. Pharmacol. 45: 2123-2127, 1993; NPC 15669 and analogs;
8). Peptide derivatives such as ACTH and analogs; soluble TNF-receptors; TNF-antibodies; interleukins, other cytokines, soluble receptors for T-cell proteins; antibodies to receptors for interleukins, other cytokines, T-cell proteins;
9. Calcipotriols as activators of the synthesis of various nerve growth factors and their analogs or growth factors themselves or promote oligodendrogenesis (or prevent their apoptosis or destruction) ), A small peptide that enhances myelin regeneration.

  The data according to the present invention show that, in terms of its effect in various animal models of human MS cells as well as demyelination (eg, various EAE models), these various novel drugs are intended to achieve good protection with few side effects It shows that they can be combined.

  “Immunodegenerative agent” refers to stimulating or inhibiting the cellular activity of cells in the immune system, eg, T-cells, B-cells, macrophages or other APC cells, for example directly or indirectly on the immune system. Means agents that act by acting on or outside components of the immune system, such as hormones, receptor agonists or antagonists and neurotransmitters that stimulate, suppress or degenerate the immune system; It may be an immunosuppressant or immunostimulant.

  By “anti-inflammatory agent” is meant an agent that treats, for example, an inflammatory response, ie, a tissue response to injury, such as an agent that treats the immune, vascular or lymphatic system.

In addition, there are many processes in this new combination treatment plan, where the assumed symptoms of MS include an initial specific or including an impaired suppressor T-cell action on myelin-specific autoimmune T-cells. Can be associated with existing or future drugs after a non-specific course (see simple schedule):
The invention also relates to a method for treating or preventing MS comprising applying an effective amount of a compound of formula I or II alone, preferably rolipram, type IV phosphodiesterase.

  The present invention, in one aspect, relates to a racemic or optically active 4- (polyalkoxyphenyl) -2-pyrrolidone of formula II that is useful in the prevention or treatment of multiple sclerosis:

［式中、Ｒ_１及びＲ_２は、それぞれ等しい又は異なるもので、少なくとも一つはメチル以外の炭素原子数１８までを有する炭化水素、ヘテロ環、又は１個以上のハロゲン原子、ヒドロキシ、カルボキシ、アルコキシ、アルコキシカルボニル、又はアミノ基で置換されている炭素原子数１〜５のアルキルであり；Ｒ’は水素原子、アルキル、アリール又はアシルであり；Ｘは酸素原子又は硫黄原子である］。

[Wherein R ₁ and R ₂ are the same or different, and at least one is a hydrocarbon having up to 18 carbon atoms other than methyl, a heterocycle, or one or more halogen atoms, hydroxy, carboxy, An alkoxy, alkoxycarbonyl, or alkyl of 1 to 5 carbon atoms substituted with an amino group; R ′ is a hydrogen atom, alkyl, aryl, or acyl; X is an oxygen atom or a sulfur atom].

  These compounds and methods for producing them are described, for example, in U.S. Pat. S. No. 4,193,926 and WO 92/02220.

  A preferred compound according to the invention is rolipram. Rolipram is 4-[(3-cyclopentyloxy) -4-methoxyphenyl] -2-pyrrolidinone. See Merck Index 11th edition, 1312-1313. This is commercially available from Schering AG, Berlin, Germany or can be produced, for example, by Belgian Patent No. 826923 or US Pat. No. 4,193,926. This is usually useful as an antidepressant, e.g. Schwab et al. Mol. Pharmacol. 12, 900 (1976); Wachtel, Neuropharmacol. 22, 267 (1983); Wachtel and H.M. Schneider, Neuropharmacol. 25, 1119 (1986); Krause and G. Kuehne, Xenobiotica 18, 561 (1988). The clinical evaluation of rolipram for depression is Zeller et al. , Pharmacopsychiatry 17, 188 (1984). Comparative clinical trials with amitriptyline (qv) in severe depression are described in Eckmann et al., Curr. Ther. Res. 43, 291 (1988). Derivatives of rolipram, ie compounds structurally related to rolipram, such as those of formula I, can also be used according to the invention and are effective in the prevention and / or treatment of MS.

  The present invention provides a type IV phosphodiesterase inhibitor, preferably a compound of formula I, especially rolipram, in multiple sclerosis (MS) to prevent the severity, symptoms and / or recurrence periodicity of the disease and / or Or it is used for amelioration, for example, to extend the time interval of a disease state in which symptoms are acute and / or to suppress the progressive or autoimmune response associated with the disease.

  The present invention therefore relates to the application of such compounds to patients for the prevention or treatment of MS, for example of an effective amount of formula I or II, preferably rolipram. The applied amount of said compound, for example rolipram, is used to prevent or ameliorate the symptoms or recurrence of the disease or affect the final course of the disease, eg inflammatory response in the brain, inflammatory lesions Effective to block the typical symptoms associated with the expression, neuronal or glial cell death and / or demyelination and the pathogenesis of the disease.

  The present invention provides a pharmaceutical composition useful for the prevention or treatment of multiple sclerosis comprising a compound of formula I or II, preferably a type IV phosphodiesterase inhibitor, preferably rolipram or a combination of such agents. Also provide. According to this method, a compound of formula I or II or a drug mixture can be applied, for example, in a single dose, in multiple doses, for example by intradermal injection or by sustained release means such as an implanted osmotic pump.

  According to the present invention, a pharmaceutical composition of formula I or II or a composition such as described above containing an effective amount of each of the compounds described is treated with multiple sclerosis, eg various multiple sclerosis metastases such as optic neuromyelitis. It can be applied to patients with (Devi's disease), diffuse sclerosis, metastatic sclerosis, acute diffuse encephalomyelitis and optic neuritis.

  The syndromes of MS that are prevented, ameliorated, or treated include: weakening and / or paralysis of one or more limbs; extreme tight band-like sensations around the body or limbs Irritation; fatigue or weak control of one or both legs, leading to convulsions or ataxic palsy; superactive tendon reflexes; disappearance of abdominal reflexes; Lhermitte's signs; retrobulbar or optic neuritis; Stable; brainstem symptoms (double vision, dizziness, vomiting); dysuria; half body insufficiency; trigeminal tract; other pain syndromes; nystagmus and ataxia; cerebellar-type ataxia; Charcot's 3 signs; compound eyes; Intranuclear eye pain; muscle paralysis or sensory paralysis of the facial muscles; hearing loss, tinnitus; irregular hallucinations (by cochlear coupling); dizziness and vomiting (vestibular coupling); temporary facial paralysis or trigeminal neuralgia paralysis; Happiness Depression; dementia; numbness; lower back pain; sharp, burning, slight local and lower back pain in the hands or both feet; sudden attack of nerve deficits; stuttering and ataxia; And perceptual dysfunction; flash; seizure itch; and / or stiff stroke in the form of flexion (muscular tension failure) stiffness ranging from the hands, wrists and elbows to lower limbs. Patients with MS have one or more of these symptoms or other clinical manifestations typically associated with MS, one or more can be improved by applying the compounds of the invention.

  Application of type IV phosphodiesterase inhibitors such as rolipram or the latter in combination with other agents may cause physiological and pathological deterioration associated with MS, such as inflammatory responses in other areas of the brain and nervous system, blood- Blocking or reducing brain barrier breakage, manifestation of lesions in the brain, tissue destruction, demyelination, autoimmune inflammatory response, acute or chronic inflammatory response, neuronal cell death and / or glial death it can.

  The agents of the present invention may be used in various types of MS, including multifocal, CNS, relapse and relief processes; multifocal, CNS progressive course; unilateral recurrence and relief courses; and other sclerotic metastases Useful for treatment. For example, James B. See Wingaarden, 1988, Cecil's Textbook of Medicine.

  The effects of application of rolipram and other type IV phosphodiesterase inhibitors and combinations of this with other drugs include, for example, prevention of the disease, improvement of symptoms of the disease, reduction of the annual rate of exacerbation (eg per year) Reduction in the number of pathologies of the disease), slowing the progression of the disease or reducing the occurrence of brain damage (such as identified on an MRI scan). Relapse of the disease, for example MS pathology, reduces the severity of this symptom associated with the onset of MS (symptom as described above), or if the pathogenesis is characterized by exacerbation and worsening of symptoms, The period between pathogenesis can be improved, for example, by extending days, weeks, months or years, or by preventing or delaying the development of cerebral inflammatory lesions. For example, Adams, R.M. D. , Principle of Neurology, 1993, page 777, description of neurological inflammatory lesions.

Other particularly suitable type IV phosphodiesterase inhibitors that can be used in the present invention include compound RO 20-1724 (4-[(3-butoxy-4-methoxyphenyl) methyl]-), a compound described in WO 93/19068. 2-imidazolidinone), ICI 63197 (2-amino-6-methyl-4-propyl [1,2,4] triazolo [1,5-a] pyrimidin-5 (4H) -one), denbufylline And etazolate.
“Type IV phosphodiesterase inhibitors”, “specific type IV phosphodiesterase inhibitors” and similar expressions indicate that a compound selectively binds to a type IV phosphodiesterase when compared to a known type, eg, a type I, II or III phosphodiesterase. selective like or inhibit, i.e. mean specific inhibitors, for example where the compound has a lower IC 50 _(more effective) with respect to type IV phosphodiesterase, other known, for example, I The IC ₅₀ is, for example, 2 times, 5 times, 10 times, 50 times or more effective for type IV phosphoesterases compared to type II, III or type phosphodiesterases. Such selectivity of the compounds according to the invention with respect to type IV phosphodiesterase is achieved by other means, for example in a manner that is diffused to its target, for example the compound targets a specific type of tissue or cell with type IV phosphodiesterase. A system that can be combined with a drug that does it; a system that interacts with host metabolism and / or physiologically; or activation of a PDE inhibitor is present in the desired cell or tissue but not in others It can also be provided by synthesizing PDE inhibitor precursors where achieved by the enzyme.

  Specific inhibition of type IV phosphodiesterase is usually described in, for example, Reeves et al. Biochem. J. et al. 241: 535-541, 1977; for example, Schade et al. , Europ. J. et al. Pharmacol. , 230: 9-14, 1993; or WO 93/19068. Regarding the specificity of phosphodiesterase and its measuring method, see, for example, Nicholson et al. Terends Pharmacol. Sci. 12: 19-27 (1991).

  The activity of a type IV phosphodiesterase inhibitor according to the present invention, eg rolipram, can be assayed in animals suffering from experimental allergic demyelinating encephalitis (EAE), experimental T-lymphocyte initiation disease of the CNS, for example. . This can be formed, for example, in rodents, guinea pigs, pupae and primates by immunizing animals with, for example, myelin from the human brain and / or by prolonged corticosteroid application. This can also be formed by injecting animals with T-lymphocytes obtained from EAE diseased animals.

  In particular, this activity can be assayed, for example, with Calithrix Jacks immunized with myelin from human brain. This wilt develops EAE with histopathology and neurological symptoms that are substantially similar to those in a particular stage of MS disease in a person.

  The present invention generally uses a combination of a PDE IV inhibitor and an interferon derivative, prostan derivative, glucocorticoid, immunosuppressant, lipoxygenase inhibitor, leukotriene antagonist, anti-inflammatory substance, peptide derivative or calcipoteriol or the like It relates to the treatment of MS.

  Advantageous combinations include PDE IV inhibitors and interferon derivatives, prostan derivatives or leukotriene antagonists such as betacellone, iloprost, cicaprost or 5-[(E)-(2S) -2 (1E, 3E)-(5S)- 5-hydroxy-6,6-trimethylene-9-phenyl-1,3-nonadiene-8-ynyl) -cyclohexylidene] -pentanoic acid or its ester.

  The pharmaceutical composition according to the present invention usually contains pharmaceutically commonly used substances including excipients, carriers, adjuvants and buffers (see, for example, Remington's Pharmaceutical Sciences, 18th. Mack Publishing Company (1990)). ). This composition can be applied to, for example, parenteral, enteral, oral, intramuscular, topical, intraperitoneal, intravenous, aerosol, directly into CNS cerebrospinal fluid, or implanted in mini-osmote. Can be applied by sustained release using an Ick pump (eg, ALZET pump manufactured by ALZA Corporation, PO Box 10950, Palo Alto, CA 94303) or in a manner useful to achieve other effects .

  Conventional excipients include pharmaceutically acceptable organic or inorganic support materials suitable for parenteral, enteral or topical application that do not adversely react with the drug. Suitable pharmaceutically acceptable adjuvants include water, salt solution, alcohol, gum arabic, vegetable oil, polyethylene glycol, gelatin, lactose, amylose, magnesium stearate, talc, salicylic acid, viscose paraffin, perfume oil, fatty acid monoglyceride And diglyceride, pentaerythritol fatty acid ester, hydroxy-methylcellulose, polyvinylpyrrolidone, cyclodextrin and the like, but are not limited thereto. The formulations can be sterilized and, if desired, stabilizers, wetting agents, emulsifiers, salts to affect osmotic pressure, buffering agents, coloring agents, flavoring and / or fragrance that do not adversely react with the active substance. Can be mixed with substances.

  For parenteral application, sterile solutions for injection, advantageously oily or aqueous solutions, as well as implants, including suspensions, emulsions or suppositories are particularly suitable. Ampoules are an advantageous unit application.

  For enteral application, tablets, dragees, suppositories or capsules with a hydrocarbon carrier or binder are particularly suitable. The support may be lactose, corn starch, potato starch or a mixture thereof. When a sweetened vehicle is used, syrup or electric seal can be used.

  This composition may optionally comprise pharmaceutically conventional additives such as buffers, electrolytes such as sodium chloride, antioxidants such as ascorbic acid, adjuvants such as methylcellulose, lactose and mannitol and / or surfactants, For example, lecithin and tweens and / or flavoring fragrances such as essential oils can be added to formulate as an aqueous solution.

  The amounts of the type IV phosphodiesterase inhibitor and the pharmaceutical composition can usually be determined based on the information described herein, for example using an EAE model. However, any amount effective in MS treatment can be applied to ameliorate or treat this disease. The dose can be determined in a conventional manner, for example, see Remington's Pharmaceutical Sciences, 18th ed. See Mack Pulishing Company (1990). The composition can be applied in a single application unit or multiple application units, for example 2, 3, or 4 times per day, or using an osmotically pump that continuously diffuses the drug. The type IV phosphodiesterase inhibitor can be applied simultaneously as an anti-inflammatory agent, an immunodegenerative agent, etc., in single or divided dose units, or at different times or eg continuously.

  The exact dose of the ingredient or composition to be applied will be determined by the clinician's instructions and by, for example, the ability of the compound to be applied, the patient's age, weight, symptoms and response.

  Generally, the PDE IV inhibitor alone is about 0.005 to 2 mg / kg / day, preferably 0.1 to 7 mg / kg / day, or 0.5 mg / kg / day, more preferably 0.005. In the amount of ~ 0.1 mg / kg / day and immunodegenerative or anti-inflammatory drugs etc. alone, in the amount of about 0.01 μg / kg / day for prostacyclin, or about 10 mg / kg for steroids / Day applies in amounts. According to the present invention, the latter is at a lower dose than would be expected with a simple additive effect, eg about 0.0005 mg to 0.01 mg / kg / day for a PDE IV inhibitor, and an immunodegenerative agent Or for anti-inflammatory drugs, it can be applied at about 0.001 μg / kg / day to about 1 mg / kg / day.

  Since the present invention relates to the treatment of MS using a combination of active ingredients to which the components can be applied separately, the present invention also relates to combining separate pharmaceutical compositions in kit form. This kit form is particularly advantageous when the separate components are applied in different application forms (ie oral and parenteral) or at different application intervals.

  For those skilled in the art, using the foregoing description, it is believed without difficulty that the present invention can be utilized to its fullest extent. Accordingly, the following advantageous specific embodiments are merely illustrative and are not intended to limit the remainder of this description.

In the foregoing and following examples, all temperatures are unmodified Celsius; and unless otherwise indicated, “parts” and “%” are “parts by weight” and “wt%”.
BRIEF DESCRIPTION OF THE DRAWINGS Various other objects, aspects and attendant advantages of the present invention will become more fully apparent when considered in conjunction with the accompanying drawings, wherein like-related characters are referred to throughout the drawings, wherein: Similar or identical parts are shown here:
FIG. 1 shows the prevention of experimental allergic encephalomyelitis (EAE) by rolipram in a rodent. A, B and C applied rolipram (10 mg / kg) in DMSO and D and F applied the same amount of DMSO. Rolipram or placebo was applied to rodents immunized with human spinal cord homogenate 5 days after immunization.

  FIG. 2 shows a treatment with a rolipram of a rodent with EAE.

Example 1
Rolipram is manufactured by Schering AG (Berlin) and consists of (+) and (-) racemates of 4-[(3-cyclopentyloxy) -4-methoxypentyl] -2-pyrrolidinone. This was dissolved at 20 mg / ml in dimethyl sulfoxide (DMSO).

  From autopsy material, Human brain white substance homogenate in complete Freund's adjuvant (CFA) containing Tuberculosis (H37 Ra strain) was prepared.

  Bordetella pertussis vaccine was obtained from Massachusetts Public Health Department, Biological Laboratories, Boston, Massachusetts.

  Zofran was obtained from the University of California Medical Center Pharmacy.

  Caries were obtained from the New England Regional Primate Center and raised according to the guidelines of the International Animal Care and Use Committee of the University of California, San Francisco.

Animals were killed by intradermal injection (0.6 ml) at 4 sites on the axilla and groin. Immunization with human brain white substance homogenate (200 mg) in CEA containing 3 mg / ml Tuberculosis (H37 Ra strain). On the day of immunization and again 2 days later, inactivated Bordetella pertussis 10 × 10 ¹⁰ was injected intravenously in 10 ml of saline.

  On day 5 after immunization, DMSO (placebo) or DMSO containing rolipram was injected subcutaneously into the back neck of the animals at a dose of 10 mg / kg. To prevent salivation, vomiting, excessive grooming and swinging, treatment with DMSO or rolipram-containing DMSO was preceded 20 minutes by intramuscular injection of 0.3 to 0.6 mg / kg of Zofuran (Odantron Hydrochloride, Glaxo) I let you. Such treatment was repeated every 48 hours throughout the study.

Animals were observed daily and presented with a standardized recording system for recording the severity of clinical symptoms:
0. Normal Lethargy, loss of appetite, weight loss 2. Ataxia, tremor 3. Blind, lower limb paralysis or unilateral paralysis Limb paralysis 5. Drowning.

  At different times, animals were anesthetized and subjected to MRI.

Experiment 1: Prevention of EAE by rolipram treatment The rodents were immunized with cerebrospinal homogenate as described above. On the 5th day after immunization, three rodents were applied with rolipram (10 mg / kg) in DMSO. See A, B and C in FIG. Two rodents received the same amount of DMSO at the same interval. See D and E in FIG. This treatment was repeated every 48 hours.

  Animals treated with DMSO (placebo) constantly developed clinical symptoms with EAE 15 days after immunization; see FIGS. 1D and 1E. None of the rolipram-treated animals developed symptoms during the 8 weeks of observation; see FIG. 1, A, B and C.

  Magnetic resonance imaging (MRI) analysis shows that two animals with EAE symptoms show more than one lesion in the brain, which is enhanced with maglevist (gadotolinium, DTPA) and is found in EAE or MS Active edema response consistent with (Alvord et al.). None of the rolipram-treated animals showed detectable lesions during the 8 weeks of observation.

Development of EAE after Rolipram Exclusion Sixty days after immunization, treated animals were removed from treatment and observed for signs of EAE. Two rodents began to show clinical signs of EAE 17 days after rolipram exclusion.

Experiment 2: Treatment of active EAE with rolipram Previous experiments clearly showed that rolipram was able to prevent EAE. It is important to determine whether rolipram can act on active EAE. In this experiment, the rodents were immunized as described above to develop chronic EAE symptoms and subsequently treated with rolipram. Animals were treated with increasing doses of rolipram in DMSO applied as described in previous experiments. Physiological symptoms were monitored as described and MRI analysis was performed before and after treatment (see FIG. 2).

  The animals showed a marked improvement 10 days after the start of treatment. The animals showed MRI improvement 14 days after the start of rolipram treatment, from which time the symptoms stabilized with a gradual improvement.

  The results of Experiment 1 indicate that rolipram treatment blocked EAE neurological symptoms. The MRI results indicate that this inflammatory response was blocked and demyelination did not occur. Untreated control animals clearly developed EAE and inflammatory lesions according to MRI analysis. The fact that the treated animals developed EAE when the treatment was removed indicates that the immune response to brain homogenate has adequately initiated the disease but the subsequent process in etiology has been blocked. The results of Experiment 2 show that rolipram treatment can suppress active disease.

Example 2
To assess the ability of rolipram alone or various combinations as described above to modify the autoimmune process, TNF production in vitro by MBP-specific T-cells from MS patients and Lewis rats The impact on MBP is the major major candinate antigen in MS, and T-cell mediated immunity is critical in its pathogenesis. Similar to EAE, MBP-specific T-cells in humans often recognize epitopes that are Th1-type secretory interferon (INF) -γ and TNF / LT cytotoxic and are also encephalitogenic in EAE.

  Rolipram selectively suppressed TNF production by human MBP-specific T-cell lines (TCL), either alone or in combination, in a dose-dependent manner.

Similar results were found using encephalitogenic CD4 ⁺ MBP-specific rat TCL (L1402). TNF / LT (Lymphotoxin) production as measured by cytotoxicity bioassay was suppressed at a dose range comparable to the human line. Furthermore, the suppression was stereospecific and was 55 times more effective for the (−)-enantiomer than for the (+)-enantiomer. The ED ₅₀ of (−) rolipram, (+) rolipram and single application (−)-rolipram was 20 nM, 280 nM and 1100 nM, respectively. Previous studies have shown that the inhibition of cAMP PDE by rolipram is stereospecific. In vivo and in vitro binding data with ³ H-rolipram in forebrain tissue in mice and lattes, the (−)-enantiomer demonstrated a 15-30 fold higher affinity than the (+)-enantiomer. . In this discovery line, the data in the present invention strongly suggest that rolipram suppresses TNF / LT production in human and latte autoreactive T-cells by an intracellular cAMP PDE-dependent mechanism. .

  Both TNF and LT are produced by autoreactive T-cells. CD4 + cells have been reported to be a major source of TEN in autoimmune insulinis of NOD mice. The cytokine bioassay for TNF / LT testing used here is sensitive to TNF and LT, but 200 times more sensitive to the former.

  This result of in vivo testing gave us advice for conducting EAE (aEAE) and adoptive cell transfer (tEAE) treatment experiments after active immunization in Lewis latte. When rolipram was applied to aEAE for 7-23 days as a single treatment or as various combinations, the development of neurological symptoms was completely prevented. Treatment was initiated within 6 hours of onset of symptoms with clinically apparent EAE. In the treatment group, the disease only progressed slowly, while the control developed severe EAE. None of the rolipram-treated animals developed a fourth degree (paraplegia), while the vehicle-treated control group of 4-7 animals reached the level of injury in a typical experiment. Similar effects were observed with tEAE. Prophylactic treatment results in 0.3 ± 0.11 (n = 5) in the treated group compared to 2.5 ± 2.5 (n = 5) (p <0.01) in the control. Only symptoms with a low mean maximum score (MMN) were shown. Treatment after the onset of symptoms also resulted in a significant decrease in maximum severity (treatment group: MS 0.7 ± 0.10, n = 5; vehicle treated control group: 2.45 ± 0.56, n = 5, p0.01).

  Furthermore, the drug was applied to animals from the day of active immunization to 11 days in order to distinguish between long-term preventive or temporary inhibitory effects. In this way, aEAE in treated animals was delayed by 3.5 days, but the severity and duration of the disease was similar compared to controls. This indicates that the suppression of EAE and expected TNF / LT production is temporary, and the loss of autoreactive T-cells by rolipram is not as promising as for example with cyclophosphamide. ing.

  Histological analysis was performed on selected animals with aEAE. There were only a few mild cell lesions in the prophylactically treated animals. In the controls, 2-3 animals treated after the development of clinical signs showed similar cell infiltration as in the controls.

  Previous studies have shown that inflammatory infiltration in the central nervous system does not have to be associated with the degree of neurological damage. In studies on EAE induced by myelin oligodendrocyte glycoprotein (MOG) -specific T-cells, a decrease in neurological signs was attributed to a decrease in macrophages and abnormally developed tissue inflammation, but perivascular Inflammation and synthesis of TNF, IFN-gamma and interleukin-6 were clearly present. However, the system of the present invention discusses the temporal occurrence of paralysis and the morphologically similar manifestation of infiltration in some of the treated animals against such phenomena. The present invention shows that during clinically apparent EAE rolipram treatment, the suppression of local TNF production in addition to other effects is critical regardless of any conflict with histological and clinical evaluation. Propose that there is. Using in situ hybridization, it has recently been shown that the presence of TNF-expressing cells in the CNS is associated with clinical signs in EAE. This study can be used to identify rolipram sensitive cell types in EAE and MS lesions-inflammatory cells, glial cells or both.

  In contrast, rolipram has, in its many effects, a significant amount in MS and other patients, especially in the appropriate combination, as shown in these or other experiments, as in the situations already described. Expected for therapeutic use.

  The experiment can be repeated with similar success rates by changing the reaction components and / or processing conditions of the invention generally or specifically described in the experiment.

  From the foregoing, those skilled in the art can readily ascertain the main features of the present invention, and various modifications of the present invention to adapt to various uses and conditions without departing from this spirit and scope. Can be changed and modified.

The graph which shows the result of the MRI analysis of Experiment 1. FIG. The graph which shows the result of the MRI analysis of Experiment 2. FIG.

Claims (8)

In a pharmaceutical composition for the treatment or prevention of multiple sclerosis, an interferon derivative and formula II:

[Wherein R ₁ and R ₂ are the same or different, and at least one of them is C ₁ -C ₁₈ -alkyl other than methyl, a hetero ring, or one or more halogen atoms, hydroxy, carboxy, alkoxy, And a drug thereof, which is alkoxycarbonyl or C ₁ -C ₅ -alkyl substituted with an amino group; R ′ is a hydrogen atom, alkyl, aryl, or acyl; X is an oxygen atom or a sulfur atom] A pharmaceutical composition for the treatment or prevention of multiple sclerosis, comprising an effective amount of a combination of PDE IV inhibitors that are pharmaceutically acceptable salts.   The composition according to claim 1, wherein the interferon derivative is interferon-β.   The composition according to claim 2, wherein the interferon-β is interferon-β-1b. Formula II:

[Wherein R ₁ and R ₂ are the same or different, and at least one of them is C ₁ -C ₁₈ -alkyl other than methyl, a hetero ring, or one or more halogen atoms, hydroxy, carboxy, alkoxy, And a drug thereof, which is alkoxycarbonyl or C ₁ -C ₅ -alkyl substituted with an amino group; R ′ is a hydrogen atom, alkyl, aryl, or acyl; X is an oxygen atom or a sulfur atom] Use of PDE IV inhibitors and interferon derivatives, which are pharmaceutically acceptable salts thereof, for the manufacture of pharmaceutical compositions useful for the treatment or prevention of multiple sclerosis. In a pharmaceutical composition for the treatment or prevention of multiple sclerosis, in a pharmaceutically acceptable carrier, the compound of formula II:

[Wherein R ₁ and R ₂ are the same or different, and at least one of them is C ₁ -C ₁₈ -alkyl other than methyl, a hetero ring, or one or more halogen atoms, hydroxy, carboxy, alkoxy, And a drug thereof, which is alkoxycarbonyl or C ₁ -C ₅ -alkyl substituted with an amino group; R ′ is a hydrogen atom, alkyl, aryl, or acyl; X is an oxygen atom or a sulfur atom] A pharmaceutical composition for treating or preventing multiple sclerosis, comprising an effective amount of a PDE IV inhibitor which is a pharmaceutically acceptable salt. Formula II:

[Wherein R ₁ and R ₂ are the same or different, and at least one of them is C ₁ -C ₁₈ -alkyl other than methyl, a hetero ring, or one or more halogen atoms, hydroxy, carboxy, alkoxy, And a drug thereof, which is alkoxycarbonyl or C ₁ -C ₅ -alkyl substituted with an amino group; R ′ is a hydrogen atom, alkyl, aryl, or acyl; X is an oxygen atom or a sulfur atom] Use of a PDE IV inhibitor, a pharmaceutically acceptable salt, for the manufacture of a pharmaceutical composition useful for the treatment or prevention of multiple sclerosis. In a kit containing, in a single package, a pharmaceutical composition for use in combination to treat or prevent multiple sclerosis in separate containers, the first container has Formula II:

[Wherein R ₁ and R ₂ are the same or different, and at least one of them is C ₁ -C ₁₈ -alkyl other than methyl, a hetero ring, or one or more halogen atoms, hydroxy, carboxy, alkoxy, And a drug thereof, which is alkoxycarbonyl or C ₁ -C ₅ -alkyl substituted with an amino group; R ′ is a hydrogen atom, alkyl, aryl, or acyl; X is an oxygen atom or a sulfur atom] A kit comprising a pharmaceutical formulation comprising a PDE IV inhibitor that is a pharmaceutically acceptable salt and, in a second container, comprising a pharmaceutical formulation comprising an interferon derivative.   The kit according to claim 7, wherein the interferon derivative is interferon-β.

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