IE83274B1 - Cytokine inhibitors - Google Patents

Description

COMPLETE SPECIF1CATIQN CYTOKINE INHIBITORS SMITHKLINE BEECHAM CORPORATION, a Corporation organised under the iaws of the Commonweaith of Pennsyivania, one of the United States of America, of One Frankiin Piaza, Phiiadeiphia, Pennsylvania, 19101, United States of America BACKGROUND OF THE INVENTION l5 This invention relates to the manufacture of medicaments for use in inhibiting the production of cytokines, particularly inhibiting the production of interleukin-1 and inhibiting the production of tumor necrosis factor, in a mammal, including a human.

Badger et a1., U.S. Patent No. 4,963,557 issued October 16, 1990, discloses compounds of the formula R R 1 /// 3 Formula (I) N wherein: n is 3-7; m is 1 or 2; R1 and R2 are the same or different and are selected from hydrogen or straight chain, branched chain or cyclic alkyl. provided that the total number of carbon atoms contained by R1 and R2 when taken together is 5-10; or R1 and R2 are joined together to form a cyclic alkyl group having 3-7 carbon atoms; R3 and R4 are the same or different and are selected from hydrogen or straight chain alkyl having 1-3 carbon atoms; or R3 and R4 are joined together with the nitrogen atom to form a heterocyclic group having 5-8 atoms; or a pharmaceutically acceptable salt or hydrate or solvate thereof. Badger et al., also discloses that such compounds have utility in inducing immune suppression via induction of suppressor cell like activity based on their activity in the adjuvant-induced arthritis test in rats and their activity in the suppressor cell assay. The adjuvant arthritis test is useful for detecting compounds which are inhibitors of prostanoid synthesis, disclosing or suggesting cytokine production, but is of no utility for compounds which are inhibitors of particularly compounds which are inhibitors of interleukin—1 (IL-1) and/or tumor necrosis factor (TNF). immunosuppressive compounds but is of no known utility for The suppressor cell assay is useful for detecting disclosing or suggesting compounds which are inhibitors of cytokine production, particularly compounds which are inhibitors of IL-1 and/or TNF production.

Cytokines are biological substances produced by a variety of cells, such as monocytes or macrophages. Cytokinesy affect a wide variety of cells and tissues and are important and critical inflammatory mediators of a wide variety of disease states and conditions. The inhibition of these cytokines is of benefit in controlling, reducing and alleviating many of these disease states.

Summary of the Invention This invention relates to the manufacture of medicaments for use in inhibiting the; production of cytokines, particularly inhibiting the production of inter1eukin—1 (IL-1) and inhibiting the production of tumor necrosis factor (TNF), in a mammal including a human uMngzu1cflbcuvc,cyHfldne[Hoducuoninhfififingzunountofa(xnnpound<fiWhe Fonnuh1 R1 R3 /// Formula 1 R2 (CHIN \(czi2)I1/N \ Ra wherein: n is 3-7; m is 1 or 2; R1 and R2 are the same or different and are selected from hydrogen or straight chain, branched chain or cyclic alkyl, provided that the total number of carbon atoms contained by R1 and R2 when taken together is 5-10; or R1 and R2 are joined together to form a cyclic alkyl group having 3-7 carbon atoms; A R3 and R4 are the same or different and are selected from hydrogen or straight chain alkyl having 1-3 carbon atoms; or R3 and R4 are joined together with the nitrogen atom to form a heterocyclic group having 5-8 atoms; or a pharmaceutically acceptable salt or hydrate or solvate thereof.

The discovery of a compound which inhibits cytokine production provides a therapeutic approach for diseases in which excessive or unregulated cytokine production is implicated.

Detailed Description of the Invention The preparation of all compounds of Formula (I) and pharmaceutically acceptable salts, hydrates and solvates thereof is disclosed in U.s. Patent No. 4,963,557 issued to Badger et al. on October 16, 1990 the entire disclosure of which is hereby incorporated by reference. - 4 _ By the term "cytokine" as used herein is meant any secreted polypeptide that affects the functions of other cells, and is a molecule which modulates interactions between cells in the immune or inflammatory response. A cytokine includes, but is not limited to monokines and lymphokines regardless of which cells produce them. For instance, a monokine is generally referred to as being produced and secreted by a mononuclear cell, such as a.macrophage and/or monocyte but many other cells produce monokines, such as natural killer cells, fibroblasts. basophils, neutrophils, endothelial cells, brain astrocytes, bone marrow stromal cells, epideral keratinocytes, and B- lymphocytes. Lymphokines are generally referred to as being produced by lymphocyte cells. but are not limited to, interleukin—1 (IL-1), tumor necrosis factor—a1pha (TNFa) and tumor necrosis factor beta (TNFB).

Examples of cytokines include.

By the term "cytokine production inhibiting amount" is‘ meant an effective amount of a compound of Formula (I) which will, when given for the treatment, prophylacticaly or therapeutically, of any disease state which is exacerbated or caused by excessive unregulated cytokine production, cause a decrease in the in yiyg levels of the cytokine to normal or below normal levels.

By the term "inhibiting the production of cytokines" is meant a) a decrease of excessive in yiyg cytokine levels in a mammal, including a human, to normal levels or below normal levels by inhibition of the in yigg release of cytokines by all cells, including but not limited to monocytes or macrophages; b) a down regulation, at the level of transcription or translation, of excessive in yiyg cytokine levels in a mammal, including a human, to normal levels or below normal levels; or c) a down regulation, by inhibition of the direct synthesis of a cytokines as a postranslational event.

By the term "inhibiting the production of IL-1" is meant a) a decrease of excessive in yiyg IL-1 levels in a mammal, including a human, to normal levels or below normal _ 5 _ levels by inhibition of the ip vivo release of IL-1 by all cells, including but not limited to monocytes or macrophages: b) a down regulation. at the level of transcription or translation, of excessive in vivo IL-1 levels in a mammal, including a human, to normal levels or below normal levels; or c) a down regulation, by inhibition of the direct synthesis of IL-1 as a postranslational event. meant By the term "inhibiting the production of TNF" is a) a decrease of excessive in vivo TNF levels in a mammal, including a human, to normal levels or below normal levels by inhibition of the in vivo release of TNF by all cells, including but not limited to monocytes or macrophages; b) a-down regulation, at the level of transcription or translation, of excessive in vivo TNF levels in a mammal, including a human, to normal levels or below normal levels; or c) a down regulation, by inhibition of the direct synthesis of TNF as a postranslational event.

As TNF-B (also known as lymphotoxin) has close structural homology with TNF-a (also known as cachectin) and since each induces similar biologic responses and binds to the same cellular receptor, both TNF—a and TNF—B are inhibited by the compounds of the present invention and thus are herein referred to collectively as "TNF" unless specifically delineated otherwise.

Studies have indicated that TNF is a serum glycoprotein and that its activity is associated with a high molecular weight components.

Mouse and rabbit TNF have been isolated, as has human TNF which sequence is taught in US Patent 4,879,226, issued November 7, 1989.

TNF is synthesized as a prohormone and subsequently cleaved at several sites to yield the mature hormone.

While the active polypeptide itself has been evaluated for treatment of tumors due to its earlier reported antineoplastic activity, this administration has not been without many severe toxicities. overproduction of TNF has further been implicated in the pathogenesis of endotoxin/septic shock.

See, e.g., Carswell et al., Proc. Natl. Acad. Sci. USA, _ 5 _ 1;, 3666-3670 (1975). Endotoxin comprises the lipolysaccharide component of the cell wall of gram—negative bacteria, and is a macrophage activator which induces the synthesis and secretion of cytokines and other biologically active molecules such as TNF. In sepsis, TNF production leads to hypotension, vascular endothelial permeability, and organ damage, i.e., some of the results of endotoxic shock. Adult Respiratory Distress Syndrome (ARDS) is frequently associated with sepsis and multiple organ failure which has led to the suggestion of a role for TNF in the pathogenesis of ARDS. TNF is also the agent responsible for the weight loss (cachexia) found in chronic catabolic disease states, such as long term parasitic and viral infections, and in malignancies. This weight loss is a handicap to recovery and may even be fatal.

TNF also appears to play a role as an early product in the inflammatory response. See, e.g., Old, Nature, 330, 602-03 (1987). It further appears that among the cytokines, while TNF production precedes and augments the function of IL-1 and other cytokines there is no clear data on how the relationship among these molecules contributes to inflammation-related disease TNF activates macrophages and enhances their cytotoxic TNF has been shown to be chemotactic for states. potential ig yitrg. monocytes, suggesting that the production of TNF at sites of injury may function to recruit additional macrophages and activate those macrophages already present.

Among the various mammalian conditions for which TNF is implicated in mediating or exacerbating are rheumatiod arthritis, rheumatiod spondylitis, osteoarthritis, gouty arthritis and other arthritic conditions; sepsis, septic shock, » endotoxic shock, gram negative sepsis, toxic shock syndrome, adult respiratory distress syndrome, malaria, pulmonary inflammatory disease, bone resorption diseases, reperfusion injury, graft vs. host reaction, fever and myalgias due to infection, such as influenza, cachexia secondary to infection or malignancy, cachexia secondary to acquired immune deficiency syndrome (AIDS), AIDS, keloid formation, scar tissue formation, Crohn's disease, ulcerative colitis, or pyresis.

The human acquired immune deficiency syndrome (AIDS) results from the infection of lymphocytes, and perhaps macrophages, with Human Immunodeficiency Virus (HIV). At least three types or strains of HIV have been identified, i.e., HIV-1, HIV-2 and HIV-3.

T-cell mediated immunity is impaired and infected individuals As a consequence of HIV infection, manifest severe opportunistic infections and/or unusual neoplasms. There is a continuing peed for agents which are useful in inhibiting further disease progress in an already infected individual. TNF has been implicated in various roles with the AIDS virus as described below.

Clouse et al., J. Immunol., 15;, 431 (1989), discuss that monokines secreted by activated human monocytes induced elevated levels of HIV expression in a chronically infected human T cell clone. The monokine involved in this process was identified as TNFa.

Gowda et al., J . Immunol., 42, 773 (1989), discuss that T cell activation is required for HIV entry and HIV—dependent cell fusion.

Zagury et al., Science, ggi, 850 (1986), discuss that T cell activation is required for HIV gene expression.

Wright et al., J. Immunol., lgl, 99 (1988), discuss that monocytes from HIV—infected patients produced large amounts of TNFa and interleukin—1 (IL-1 hereinafter) upon culturing ip yiggg.

Beutler et al., Nature (London), 316, 552-554 (1985), discuss the role of TNFa in cachexia.

Chiebowski et al., Nutr. Cancer, 1, 85 (1985), discuss HIV-associated states of cachexia and muscle degradation.

Wright_et al., J. Immunol. 14l(l):99-104 (1988) suggests a possible role for TNF in AIDS cachexia by elevated serum TNF and high levels of spontaneous TNF production in peripheral blood monocytes from patients.

Yale University, European Patent Application Publication Number 0,230,574 A2, published August 6, 1987, claims a method for producing pharmaceutical compositions for treating patients infected with LAV/HTLV III virus wherein such composition contains a compound which inhibits the production and/or the activity of mononuclear cell derived cytotoxic factors, such as lymphotokin, tumor necrosis factor, leukoregulin and natural killer cytotoxic factor.

It is concluded from the above references that compounds which inhibit the production of TNF will have a therapeutic effect on the treatment of acquired immune deficiency syndrome (AIDS) and/or the treatment of AIDS related complications.

Interleukin—1 (IL-1) has been demonstrated to mediate a variety of biological activities thought to be important in immunoregulation and other physiological conditions such as inflammation [See, e.g. Dinarello et al., Rev. Infect Disease, g, 51 (1984)]. The myriad of known biological activities of IL-1 include the activation of T helper cells, induction of fever, stimulation of prostaglandin or collagenase production, neutrophil chemotaxis, induction of acute phase proteins and the suppression of plasma iron levels. Specifically, there are several disease states in which excessive or unregulated IL-1 production by monocytes and/or macrophages is implicated in exacerbating and/or causing the disease. These include rheumatoid arthritis [See, e.g., Fontana et al., Arthritis ghgum, gg, 49-53 (1982)]; osteoarthritis [See, e.g., wood et al., Arthritis Rheum. gg, 975 (1983)]; toxic shock syndrome [See, e.g., Ikejima and Dinarello, J. Leukocyte Biology, g1, 714 (1985)]; other acute or chronic inflammatory disease states such as the inflammatory reaction induced by endotoxin [See, e.g., Habicht and Beck, J. Leukocyte Biology, g1, 709 (1985)]; and other chronic inflammatory disease states such as tuberculosis. [See, e.g,, Chesque et al., J. Leukocyte Biology, g1, 690 (1985)]. Benjamin et a1., "Annual Reports in Medicinal Chemistry, gg", Chapter 18, pages 173-183 (1985), Academic Press, Inc., disclose that excessive IL-1 production is implicated in: psoriatic arthritis, Reiter's syndrome, rheumatoid arthritis, osteoarthritis, gout, traumatic arthritis, rubella arthritis, and acure synovitis. (1985), reviews the biological activities which have been attributed to IL-1 and such activities are summarized in Table A.

TABLE A Biological Activities Attributed to IL-1 Fever (in rabbits, mice and rats) Hypoferremia Hypozincemia Hypercupremia Increased V Blood neutrophils Hepatic acute—phase proteins Bone resorption, including; osteoprosis and Paget's disease Cartilage breakdown Muscle proteolysis S1ow—wave sleep Endothelial procoagulant Chondrocyte proteases Synovial collagenase Endothelial neutrophil adherence Neutrophil degranulation Neutrophil superoxide Interferon production Proliferation of Fibroblasts Glial cells Mesangial cells Synovial fibroblasts EBV B—cell lines Chemotaxis of Monocytes Neutrophils Lymphocytes Stimulation of PGE Hypothalamus Cortex Skeletal muscle Dermal fibroblast Chondrocyte in Macrophage/monocyte Endothelium (PGI2) Decreased Hepatic albumin synthesis Appetite Brain binding of opioids Augmentation of T—cell responses B—cell responses NK activity IL-2 production Lymphokine production.

The discovery of a compound which inhibits Il—1 production provides a therapeutic approach for diseases in which excessive or unregulated I1-1 production is implicated.

It has now been discovered that compounds of Formula (I) and pharmaceutically acceptable salts or hydrates or solvates thereof are useful for inhibiting cytokine production in a mammals, including humans, in need of such inhibition.

An effective, cytokine production inhibiting amount of a compound of Formula (I) or a pharmaceutically acceptable salt or hydrate or solvate thereof is useful in treating, prophlactically or thereapeutically, any disease state in a mammal, including a human, which is exacerbated or caused by excessive or unregulated cytokine production. Preferably, the inhibited cytokines are IL-1 and TNF. Preferably, the disease state is selected from; increased bone resorption, endotoxic shock, cachexia secondary to acquired immune deficiency syndrome (AIDS), AIDS or malaria. Particularly preferred is the disease state of increased bone resorption, including osteoporosis and Paget's disease.

This invention relates to a method of inhibiting the production of cytokines, particularly inhibiting the production of IL-1 and TNF, in a mammal, including a human, in need thereof which comprises administering an effective, cytokine production inhibiting amount of a compound of Formula (I) or a pharmaceutically acceptable salt or hydrate or solvate thereof. A compound of Formula (I) or a pharmaceutically acceptable salt or hydrate or solvate thereof can be administered to such mammal, including a human, in a conventional dosage form prepared by combining a compound of Formula (I), or a pharmaceutically acceptable salt or hydrate or solvate thereof, with a conventional pharmaceutically acceptable carrier or diluent according to known techniques, such as those described in Badger et al. U.S. Patent No. 4,963,557 issued on October 16, 1990.

It will be recognized by one of skill in the art that the form and character of the pharmaceutically acceptable carrier or diluent is dictated by the amount of active ingredient with which it is to be combined, the route of administration and other well-known variables. A compound of Formula (I) or a pharmaceutically acceptable salt or hydrate or solvate thereof is administered to a mammal, including a human, in need of inhibition of cytokine production in an amount l sufficient to inhibit such excessive cytokine production to the extent that it is regulated down to normal levels. The route of administration may be oral, parenteral or topical. The term parenteral as used herein includes intravenous, intramuscular. subcutaneous, intranasal, intrarectal, intravaginal or intraperitoneal administration. The subscutaneous and intramuscular forms of parenteral administration are generally preferred. The daily oral dosage regimen will preferably be from about 0.1 to about 1000 mg/kilogram of total body weight.

The daily parenteral dosage regimen will preferably be from about 0.1 to about 800 mg per kilogram (kg) of total body weight, most preferably from about 1 to about 100 mg/kg. The daily topical dosage regimen will preferably be from about 1 mg to about 100 mg per site of administration. It will be recognized by one of skill in the art that the optimal quantity and spacing of individual dosages of a compound of Formula (I) or a pharmaceutically acceptable salt or hydrate or solvate thereof will be determined by the nature and extent of the condition being treated, the form, route and site of administration, and the particular patient being treated, and that such optimums can be determined by conventional It will also be appreciated by one of skill in the techniques. art that the optimal course of treatment, i.e., the number of doses of a compound of Formula (I) or a pharmaceutically acceptable salt or hydrate or solvate thereof given per day for a defined number of days, can be ascertained by those skilled in the art using conventional course of treatment determination tests.

Without further elaboration, it is believed that one skilled in the art can, using the preceding description, utilize the present invention to its fullest extent.

As used herein, the term "compound 1" refers to a compound of Formula (I) where R1 and R2 are propyl, R3 and R4 are methyl, m is 1 and n is 3 which is N,N—dimethyl— ,8-dipropyl—2-azaspiro[4,5]decane—2—propanamine.

MEASUREMENT OF IN VIVO CYTOKINE ACTIVITY Lipopolysalcharide stimulated macrophages from adjuvant arthritic rats treated with compound I produce 50% less TNF than untreated controls.

Compound 1 demonstrated a positive in yiyg response of about 75% reduction in levels of IL-1 in the above assay.

Claims (10)

Claims:

1. Use of a compound of the Formula (I) 1 R3 /// Formula 1 (CH ).N N 2 7-m \(CH2)% n \ Ra wherein: n is 3-7; m is 1 or 2; R1 and R2 selected from hydrogen or straight chain, branched chain or cyclic alkyl, provided that the total number of are the same or different and are carbon atoms contained by R1 and R2 when taken together is 5-10; cyclic alkyl group having 3-7 carbon atoms; or R1 and R2 are joined together to form a Rf and R are the same or different and are selectgd from4hydrogen or straight chain alkyl having 1- 3 carbon atoms; or R3 and R4 are joined together with the nitrogen to form a heterocyclic group having S-8 atoms; or a pharmaceutically acceptable salt or hydrate or solvate thereof; in the manufacture of a medicament for use in inhibiting the production of cytokines in a mammal, including a human.

2. A use according to claim 1 wherein the compound is N,N-dimethyl-8,8—dipropyl azaspiro[4,Sldecane-2—propanamine dihydrochloride.

3. A use according to claim 1 wherein the compound is N,N-diethyl-8,8-dipropyl azaspiro[4,5]decanepropanamine dihydrochloride.

4. A use according to claim 1 wherein the compound is administered orally.

5. A use according to claim 4 wherein from about 1 to about 2000 mg of compound are administered per day.

6. A use according to claim 1 wherein the compound is administered parenterally.

7. A use according to claim l wnerein the compound is N,N—diethyl-8,8—dipropyl azaspiro[4,5]decane—2~propanamine; or a pharmaceutically acceptable salt or hydrate or solvate thereof.

8. A use according to claim 1 wherein the mammal is afflicted with a bone resorption disease.

9. A use according to claim 8 wherein the bone resorption disease is osteoporosis.

10. therapeutic effect is the inhibition of prostaglandin A use according to claim 1 wherein the desired production.