USOO5620964A United States Patent (19) 11) Patent Number: 5,620,964 Roth et al. 45) Date of Patent: Apr. 15, 1997 54) COMPOSITIONS FORTREATING AND Podolsky, J. of Biological Chemistry vol. 260, No. 14, pp. NHBITING GASTRIC AND DUODENAL 8262-82.71. ULCERS Evans, et al., Infection and Immunity(1988) 56: 2896–2906. Evans, et al., Infection and Immunity(1989) 57:2272-2278. 75 Inventors: Stephen Roth, Gladwyne; Edward J. Lingwood, et al., The Lancet(1989) 2:238-241. McGuire, Furlong, both of Pa.; Dennis H. Langer, Princeton, N.J. (List continued on next page.) 73) Assignee: Neose Technologies, Inc., Horsham, Pa. Primary Examiner-John W. Rollins Assistant Examiner-Francisco C. Prats (21) Appl. No.: 461,000 (57) ABSTRACT (22 Filed: Jun. 5, 1995 A composition for treating and/or inhibiting gastric and duodenal ulcers, comprising an oligosaccharide selected Related U.S. Application Data from the group consisting of Formula I 63 Continuation of Ser. No. 104.483, Jul. 28, 1993, abandoned, OHO-W" O-W which is a continuation-in-part of Ser. No. 922,519, Jul. 31, 1992, abandoned. O H. O (51) Int. Cl. .................................... A61K 3/715 W-O O O-Y; 52 U.S. Cl. ................................ 514/53; 514/42, 514/54; OH X 514/58; 514/61; 514/925; 514/926; 514/927 O -W" Formula II 58) Field of Search .................................. 514/42, 53, 58, H - O O-W 514/61, 925,926,927, 54 O O 56) References Cited W-O O O-Y OH HO X U.S. PATENT DOCUMENTS or a mixture thereof; wherein: 4,935,406 6/1990 Colmena et al. ......................... 514/54 4,938,967 7/1990 Newton et al. .. ... 424/458 X is independently OH or NHAc; 5,318,780 6/1994 Wiegas et al. ........................... 424/427 Y is independently H, or an amino acid or a peptide of OTHER PUBLICATIONS 2-100, preferably 2-20, amino acids; and Boren et al., Science, vol. 262, 1892-1895 17 Dec. 1993. W, W, and W" are each independently H or Evans et al., Journal of Bacteriology, Feb. 1993, pp. 674-683, vol. 15, No. 3. HO Saitoh et al., Federation of European Biochemical Societies, May 1991, vol. 282, No. 2 385-387. O CO2Z; Lingwood, et al., Infection and Immunity, Jul. 1993, pp. HO 2 2474-2478, vol. 61, No. 6. Fauchere, et al., Microbial Pathogenesis 1990:9: 427-439. Lingwood, et al., Infection and Immunity, Jun. 1992, pp. where Z is independently H or a pharmaceutically 2470-2474, vol. 60, No. 6. acceptable cation, and Robinson, et al., J. Med. Microbiol., vol. 33 (1990). P is independently H or 277-284, 1990 The Pathological Society of Great Britain and Ireland. HO Figueroa, et al., Journal of Infection (1992) 24,263-267. HO Talley, et al., Journal of the National Cancer Institute (1991) AcNH O COZ 83, 1734-1739. HO Wotherspoon, et al., The Lancet, vol. 342, Sep. 4, 1993, pp. 575-577. OH Russell, et al., The Lancet, vol. 342, Sep. 4, 1993, pp. 571-574. where Z is defined as above; National Institutes of Health, Concensus Development Con wherein at least one of W, W" or W" is an O-N- ference Statement, Helicobacter pylori in Peptic Ulcer Dis acetylneuraminic acid moiety, and ease, Feb. 7–9, 1994, 19 pages. wherein W" and W" are not simultaneously o-N-acetyl Falk, et al., Proc. Natl. Acad. Sci, USA, vol. 90 pp. neuraminic acid moiety 2035-2039, Mar. 1993. with the proviso that the compound of Formula II is not Huang, et al., Journal of General Microbiology (1992), 138, NAN o(2-3)Gal B1-4 Glu or NAN O(2-6)Gal 1503-153. 31-4 Glu; and a kit for detecting the presence of Koybayashi et al., Infection and Immunity, Oct. 1993, pp. Helicobacter pylori comprising at least one com 4058-4063. vol. 61, No. 10. pound of the Formula I or II, are described. Clyde, et al., Infection and Immunity, Oct. 1993, pp. 4058-4063, vol. 61, No. 10. 19 Claims, No Drawings 5,620,964 Page 2 OTHER PUBLICATIONS Mizuochi et al., J. Biological Chemistry, 255(8):3526–3531, (1980). Tzouvelekis, et al., Infection and Immunity(1991) 59.4252-42.54. Kessler et al., J. Biological Chemistry, 254(16):7909-7914, Lambert, Reviews of Infectious Diseases(1991) 13 (1979). (Suppl.8):S691-5. Spiro et al., J. Biological Chemistry, 249(18):5704-5717, Dunn, et al., Reviews of Infectious Diseases(1991) 13 (Suppl.8):S657-64. (1974). Hayes et al, J. Biological Chemistry, 254(18):8777-8780, Pelczar et al, Elements of Microbiology, McGraw-Hill (1979). Book Company:New York, 1981, pp. 222-223. 5,620,964 1. 2 COMPOSITIONS FOR TREATING AND was isolated from red blood cells, and mucosal scrapings of NHIBITING GASTRIC AND DUODENAL pig stomach and human stomach. The investigators postu ULCERS lated that the material was a sulphated alkylacylglycero lipid, but the actual structure of this material was not been reported. Subsequent investigations (Lingwood et al., Infec RELATED APPLICATIONS tion and Immunity (1992) 60:2470-2474) showed that this This application is a continuation of application Ser. No. receptor is phospatidylethanolamine. 08/104.483, filed on Jul. 28, 1993, abandoned, which is a Tzovelekis et al (Infection and Immunity (1991) Continuation-in-Part of Ser. No. 07/922,519, filed on Jul. 31, 59:4252-4253) reported binding inhibition of H. pylori to 1992, abandoned. 10 HEp-2 cells by gastric mucin. The investigators observed that purified mucin showed the greatest inhibition of H. BACKGROUND OF THE INVENTION pylori binding while asialomucin exhibits somewhat dimin ished inhibition and periodate-oxidized mucin exhibited the 1. Field of the Invention lowest level of binding. On these observations, the research The present invention relates to compounds and compo ers concluded that sialic acids are at least partially respon sitions for treating and inhibiting gastric and duodenal 15 sible for the binding interaction between H. pylori and ulcers, and to methods of treating and inhibiting gastric and human gastric mucin. duodenal ulcers. Thus the binding inhibition studies all point to a H. pylori 2. Discussion of the Background binding specific receptor which possesses an N-acetyl Infection by the gram-negative, spiral, microaerophilic 20 neuraminic acid (sialic acid) (Tzouvelekis et all and Evans et bacterium Helicobacter pylori (H. pylori), formerly known al) bound in a 2->3 manner to a lactose (Evans et al). as Campylobactor pylori (C. pylori), is a primary cause of In addition to the numerous binding inhibition studies, non-autoimmune gastritis, is a factor in peptic ulcer disease methods have been pursued to treat gastric and duodenal and is more common in patients with gastric carcinoma. ulcer patients. Colloidal bismuth subcitrate (CBS) has been First isolated by Warren (Lancet (1983) 1:1273) and Marshal 25 used successfully in treating both gastric and duodenal ulcer (Lancet (1983) 1:1273-5), H. pylori has been isolated in diseases (for a review, see Lambert in Reviews of Infectious gastric tissue biopsies in patients throughout the world. Diseases (1991) 13 (Suppl. 8):S691-5. CBS has proven While the precise mechanism of inflammation is not well effective as a histamine H2 antagonist and has been asso understood, H. pylori is found in association with the apical ciated with lower relapse rates after cessation of therapy surfaces of gastric mucous-secreting cells. 30 attributed to CBS's ability to eradicate H. pylori. Bismuth Due to the site specificity of attachment, it has been subsalicylate (BSS) has also been observed to inhibit H. suggested that there are specific attachment sites for H. pylori. pylori which exist on gastric and duodenal mucous-secreting Additional studies in eliminating H. pylori have been cells. Numerous studies have been undertaken to attempt to conducted using the proton pump inhibitor, omeprazole. identify the specific binding site of H. pylori. 35 Coleman et al (U.S. Pat. No. 4,935,406) reported a Evans et al (Infection and Immunity (1988) method for relieving gastrointestinal disorder, resulting from 56:2896–2906) reported that H. pylori binding to an eryth H. pylori population, through the administration of bismuth rocyte receptor is preferentially inhibited by N-acetyl (phosph/sulfated saccharide compositions. The saccharide neuraminyl-o.(2->3)-Gal 1-4 GlcNeuAc(2->3)-lactose) compositions according to this method are simple phos as compared with NeuAc(2->6)-lactose. Sialoproteins 40 phates and sulfates of aldose and ketose monosaccharides. which contain the NeuAc(2-3)Gal isomer of NeuAc-lac Clinical trials have been reported (Evans et al, Ann. tose, i.e., human erythrocyte glycophorin A, fetuin, and Internal Med. (1991) Aug. 15, 115(4):266-9) in treating H. human o-macroglobulin, also inhibited H. pylori binding, pylori using ranitidine in conjunction with a "triple therapy' but at much higher concentrations (mg/ml) than that of amoxicillin or tetracycline, metronidazole (an antiproto observed for NeuAc(2->3)-lactose, while no inhibition was 45 zoal), and BSS. The clinical studies suggested that ulcer observed for the corresponding asialoproteins. healing was more rapid in patients receiving ranitidine plus These researchers further observed that NeuAc-lactose the "triple therapy” than in patients receiving ranitidine (also called sialyllactose) containing primarily the alone. NeuAc(2-6)Gal isomer showed no inhibition of binding, However, long-term eradication of this organism has been leading the researchers to conclude that the receptor on the 50 difficult with these therapies. The antibiotic approach runs erythrocytes is a sialoprotein containing NeuAc(2->3)Gal. the risk of the development of new antibiotic resistant Although the NeuAc(2-3)Gal moiety, which Evans et al strains.