USOO61239.43A United States Patent (19) 11 Patent Number: 6,123,943 Baba et al. (45) Date of Patent: Sep. 26, 2000 54 NF-KBACTIVITY INHIBITOR 8-301761 11/1996 Japan ............................. A61K 31/47 75 Inventors: Masanori Baba, Kagoshima; Minoru OTHER PUBLICATIONS Ono, Tokyo, both of Japan Sato et al. Eur. J. Pharmacol. (1982) 83: 91-95. 73 Assignee: Kaken Shoyaku Co., Ltd., Tokyo, Japan Primary Examiner Jean C. Witz Attorney, Agent, or Firm Sughrue, Mion, Zinn, Macpeak 21 Appl. No.: 09/037,712 & Seas, PLLC 22 Filed: Mar 10, 1998 57 ABSTRACT 30 Foreign Application Priority Data This invention relates to an NF-kB activity inhibitor which contains alkaloids originated from a plant belonging to the Dec. 22, 1997 JP Japan .................................... 9-353879 genus Stephania of the family Menspermaceae, derivatives thereof and Salts thereof, as the active components, to an (51) Int. Cl." ..................................................... A61K 35/78 agent for use in the treatment and prevention of diseases 52 U.S. Cl. ....................... 424/195.1; 514/308; 514/387; upon which the NF-kB activity inhibiting action is effective 514/415 and to an inhibitor of the expression of related genes. Since 58 Field of Search ......................... 424/195.1; 514/387, Said active components exert an action to inhibit transcrip 514/308, 415; 546/140,139; 534/790 tion of DNA having an NF-kB recognition sequence by inhibiting the activity of NF-kB, the drug of the present 56) References Cited invention can inhibit expression of genes of certain Sub U.S. PATENT DOCUMENTS stances Such as cytokines, inflammatory cytokine receptor antagonists, MHC class I, MHC class II, B2 microglobulin, 5,025,020 6/1991 Van Dyke ............................... 514/280 5,534,523 7/1996 Ono et al. ............................... 514/308 immunoglobulin light chain, Serum amyloid A, 5,627,195 5/1997 Hu ................ ... 514/321 angiotensinogen, complement B, complement C4, C-myc 5,641,773 6/1997 Pardee et al. ........................... 514/221 gene, HIV, SV40, CMV, adenovirus and the like, so that the inventive drug is useful in treating and/or preventing various FOREIGN PATENT DOCUMENTS diseases in which these Substances are taking roles. 0 476 391 A2 3/1992 European Pat. Off. ... A61K 31f47 1-233220 9/1989 Japan. 6 Claims, No Drawings 6,123,943 1 2 NF-KBACTIVITY INHIBITOR genus Stephania of the family Menspermaceae, as its active ingredient, inhibits production of TNFC, interleukin-6 (to be FIELD OF THE INVENTION referred to as IL-6 hereinafter in Some cases) and This invention relates to an NF-kB activity inhibitor, to interleukin-8 (to be referred to as IL-8 hereinafter in some agents for use in the treatment and prevention of diseases cases) (JP-A-8-301761, the term “JP-A” as used herein upon which NF-kB activity inhibiting action is effective, means an “unexamined published Japanese patent particularly an agent for the treatment and prevention of application'). inflammatory diseases, an agent for the treatment and pre Phospholipid which constitutes the biological membrane vention of autoimmune diseases and an agent for the treat releases arachidonic acid by the action of phospholipase A. ment and prevention of Viral diseases, and to a gene expres Leukotriene, thromboxane, prostaglandine and the like are Sion inhibitor. produced from the arachidonic acid by the action of 5-lipoxygenase or cyclooxygenase. These Substances exert BACKGROUND OF THE INVENTION complex physiological activities and take important roles in DNA as the Substance of genes is regulated by various the maintenance and regulation of the living body. In the factors, and expression of its genetic information is con 15 living body, various cytokines are released by receiving trolled thereby. For example, transcription of genetic infor various types of Stimulation and cause inflammatory reac mation from DNA to RNA is controlled by a plurality of tions. The prior art drugs inhibit expression of histamine and DNA binding proteins which recognizes Several to Scores of leukotriene B4 or prostaglandine E2 or the like inflamma nucleotide Sequences on the gene and bind thereto. NF-KB tory protein by the antagonism on mediator receptors of (nuclear factor-KB) known as one of such DNA binding histamine and the like or by the inhibition of lipoxygenase, proteins is present in the nuclear extract of B cells which are cyclooxygenase and the like metabolic enzymes in the antibody-producing cells and has been identified as a factor arachidonic acid cascade. However, effects of non-Steroidal that binds to the enhancer of immunoglobulin K chain (Igk) drugs are expected for only Symptomatic therapy and not gene. With the progreSS of Studies on this factor, it has been Sufficient as radical therapy, while Steroid drugs are effective revealed that this is a transcription factor which takes part in 25 but have a problem in that they cannot be administered for the expression induction of a large number of genes that are a prolonged period of time due to their Strong Side effects. induced by Stimulation and is broadly concerned in the Particularly, autoimmune disease and the like inflammatory regulation of Vital phenomenon. diseases become chronic in many cases and therefore require This NF-kB is generally present in the cytoplasm in the prolonged medical treatments, So that drugs having Side form of a complex in which its homodimer of proteins effects are not applicable to Such diseases. In addition, having a molecular weight of 50 kD or its heterodimer of a NF-kB takes an important role in the replication of HIV-1, protein of 50 kD in molecular weight and a protein of 65 kD so that search for a substance capable of inhibiting NF-KB in molecular weight is bonded to a protein called I-KB which activity is expected for not only its anti-inflammatory effects inhibits activity of the dimer. When a certain stimulation is but also inhibition of acquired immunodeficiency Syndrome given to the cells, I-KB is modified and released from the 35 (AIDS and the like) by its effect to inhibit transcription of complex to cause activation of NF-kB, so that the dimer is long terminal repeat (LTR) of HIV-1, namely replication of transferred into the nucleus and its DNA binding activity the virus. becomes detectable. It is known that this activity is gener SUMMARY OF THE INVENTION ated as a result of direct activation, not mediated by the 40 expression of other genes Such as Second messengers and the In view of the above, it therefore becomes an object of the like. present invention to provide an NF-kB activity inhibitor. In addition, the NF-kB binding sequence on DNA has Another object of the present invention is to provide an been found in various genes and it has been shown that it is agent for the treatment and prevention of diseases upon actually important for the expression of the function of 45 which NF-kB activity inhibiting action is effective. Sill genes. The binding sequence of NF-kB (KB motif) is com another object of the present invention is to provide an posed of about 10 bases having a common Sequence which inhibitor of the expression of genes based on the NF-KB Starts with a cluster of G (guanine) and ends with a cluster activity inhibiting action. of C (cytosine) (consensus sequence 5'-GGGRNNYCCC The inventors of the present invention have conducted 3')(SEQ ID NO:1. However, a number of sequences to 50 intensive Studies on the methods and Substances which can which DNA binding proteins can be bonded are present on radically inhibit various inflammatory cytokines and found the genes of interleukin-1 (to be referred to as IL-1 herein as the results that alkaloids originated from a plant belong after in Some cases) and tumor necrosis factor (to be referred ing to the genus Stephania of the family Menspermaceae, to as TNF hereinafter in some cases) which are known as derivatives thereof and salts thereof can inhibit various inflammatory proteins, and it is known that the NF-KB 55 cytokines and the like at the gene level based on their binding sequence is also present therein (Clark, B. D. et al., transcription factor NF-kB activity inhibiting action and Nucl. Acids Res., 14, 7898, 1984; Nedospasov, S. A. et al., have an activity to inhibit transcription of HIV-1 LTR. The Cold Spring Harb. Symp. Quant. Biol., 51, 611, 1986). present invention has been accomplished on the basis of Actually, it has been reported that the binding of NF-KB these findings. inhibits transcription to mRNA (Hiscott, J. et al., Mol. Cell. 60 Accordingly, the present invention relates to an NF-KB Biol., 13, 6231, 1993; Collart, M. A. et al., Mol. Cell. Biol., activity inhibitor which comprises, as its active ingredient, at 10, 1498, 1990). least one compound Selected from the group consisting of AS a Substance which inhibits the transcription factor of alkaloids originated from a plant belonging to the genus NF-kB, an NF-kB binding protein has been disclosed in Stephania of the family Menspermaceae, derivatives thereof European Patent 584238. 65 and Salts thereof; to an agent for the treatment and preven In addition, it has been reported that a composition which tion of diseases upon which NF-kB activity inhibiting action contains an alkaloid originated from a plant belonging to the is effective (particularly an agent for the treatment and 6,123,943 3 4 prevention of inflammatory diseases, an agent for the treat isoquinoline alkaloids, FK-3000, Sinomenine, cephamonine, ment and prevention of autoimmune diseases and an agent tannagine, cephamuline and the like morphinan alkaloids, for the treatment and prevention of viral diseases); and to an lastourVilline, isocorydine, corydine and the like aporphine gene expression inhibitor.