United States Patent (19) 11 Patent Number: 5,728,680 Morozov Et Al

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United States Patent (19) 11 Patent Number: 5,728,680 Morozov Et Al III USOO572868OA United States Patent (19) 11 Patent Number: 5,728,680 Morozov et al. 45 Date of Patent: *Mar. 17, 1998 54 METHODS FOR NORMALZNG NUMBERS 4,517,119 5/1985 Felix et al. OF LYMPHOCYTES 4,526,717 7/1985 Seipke et al., 4,571,336 2/1986 Houck et al. 4.579,840 4/1986 Hahn. (75) Inventors: Vyacheslav G. Morozov, Vladimir Kh. 4.599,231 7/1986 Milich et al.. Khavinson, both of St. Petersburg, 4,612,365 971986 Birr et al. Russian Federation 4,614,731 9/1986 Horecker . 4,621,135 11/1986 Trainin et al.. 73) Assignee: Cytoven J.V. Kirkland, Wash. 4,634,682 1/987 Erickson et al. 4,659,694 4/1987 Horecker. * Notice: The term of this patent shall not extend 4,696,915 9/1987 Horecker. beyond the expiration date of Pat. No. 4,699,898 10/1987 Gottlieb. 5.538,951. 4,711952 12/1987 Kasafirek et al. 4,722.999 2/1988 Handschumacher et al. 4,751.216 6/1988 Gottlieb. 21 Appl. No.: 452,411 4,752,602 6/1988 Lipsky et al. 22 Filed: May 26, 1995 4,814434 3/1989 Goldfarb . 4,826,680 5/1989 Jaeger. Related U.S. Application Data 4,904,643 2/1990 Brunetti et al. 4,910.296 3/1990 Birr et al., 63 Continuation-in-part of Ser. No. 337,341, Nov. 10, 1994, 4.946,945 8/1990 Wojdani. Pat. No. 5,538,951, and a continuation-in-part of Ser. No. 4,983,387 1/1991 Goldstein et al.. 278,463, Jul. 2, 1994, abandoned, which is a continuation 5,070,076 12/1991 Morozov et al. in-part of Ser. No. 257,495, Jun. 7, 1994, abandoned, which 5,538,951 7/1996 Morozov et al. ......................... 514/19 is a continuation of Ser. No. 783,518, Oct. 28, 1991, 5,569,585 10/1996 Goodwin et al. ........................... 435/6 abandoned, which is a continuation-in-part of Ser. No, 678,129, Apr. 1, 1991, abandoned, which is a continuation FOREIGN PATENT DOCUMENTS in-part of Ser. No. 415,283, filed as PCT/SU88.00255, Dec. 14, 1988, abandoned. WO 9006945 6/1990 WIPO . WO 92/17191 10/1992 WIPO 30 Foreign Application Priority Data WO 93,08815 5/1993 WIPO Dec. 30, 1987 SU U.S.S.R. ................................ 4352833 OTHER PUBLICATIONS 51 Int. Cla. A6K 38/OS Rodionov et al, Vestn, Dermoatol. Venerol, 1:42-43, 1990. 52) U.S. Cl. ................................... 514/19:514/9; 514/11 Stites & Terr, Basic & Clinical Immunology, Prentice Hall. 58) Field of Search ..................................... 514/9, 11, 19 Norwalk, 1991, pp. 272-281. Brostoss et al. Clinical Immunol, Gower Med. Pub. London. 56 References Cited 1991, p. 30.10. U.S. PATENT DOCUMENTS van der Zeijst et al., J. Exp. Med, 1978, 147:1253-1266. Anisimov et al. (1982) Mechanisms of Ageing and Devel 3,679,787 7/1972 Bergy et al., opment, 19:245-258. 4,002,602 /1977 Goldstein. Anisimov et al. (1989) Mechanisms of Ageing and Devel 4.00,148 3/1977 Goldstein. opment, 49.245-247. 4,077.949 3/1978 Goldstein. 4,079,127 3/1978 Goldstein et al.. Demidov et al. (1991) “Effects of Thymus Preparations and 4,116,951 9/1978 Wang. Antituberculous Drugs on Immunological Reactivity and the 4,120,951 10/1978 Goldstein. Course of Tuberculous Process in Experimental Animals". 4,133,804 1/1979 Bach et al. Probl. Tuberk... 12:52-54 (Chemical Abstracts 116 (17), 4,148,788 4/1979 Wang. Abstract No. 165824Y). 4,167,557 9/1979 Goldstein. Goldstein et al. (1972) Proc. Natl. Acad. Sci. USA. 4.261,886 4/1981 Goldstein et al.. 69:1800-1803. 4.264,571 4/1981 Goldstein et al. Grigoriants et al. (1991) "Immunocorrection in the Com 4.297.276 10/1981 Goldstein et al., 4,339.427 7/1982 Goldstein et al.. bined Treatment of Patients with Osteomyelities Developing 4,353,821 10/1982 Birr et al.. Following Combined Injuries to the Maxillofacial and Cran 4,374.828 2/1983 Folkers et al. iocerebral Areas". Stomatologia (Mosk), 5:53-54 (Medline 4,377,511 3/1983 Lopukhin et al.. Abstract No. 92188368). 4,388,234 6/1983 Horecker . 4.389.343 6/1983 Horecker. (List continued on next page.) 4,395,404 7/1983 Low et al. 4,396,605 8/1983 Birr. Primary Examiner-Lila Feisee 4,426,324 1/1984 Meienhofer. Assistant Examiner-Susan Ungar 4,427,783 1/1984 Newman et al. 57 ABSTRACT 4,428,938 1/1984 Kisfaludy et al.. 4,442,031 4/1984 Felix et al.. This invention provides methods for normalizing the num 4,466,918 8/1984 Birr et al.. bers of lymphocytes in animals by administering the dipep 4,470,926 9/1984 Birr et al., 4,500,450 2/1985 Seipke et al.. tide L-Glu-L-Trp. 4,504,415 3/1985 Felix et al., 4,505,853 3/1985 Goldstein et al.. 12 Claims, 8 Drawing Sheets 5,728,680 Page 2 OTHER PUBLICATIONS Belokrylov, G. et al., "Ability of some amino acids incor porated into protein to stimulate the thymus-dependent Iakovlev et al. (1991) "The Biochemical And Immunologi immune response” Eksp. Biol. Medit. 102:51-53 (1986). cal Indices in the Rehabilitation Period of the Victims of the Bespalov, V.G. et al., "Inhibiting effect of thymogen, a Accident on The Komsomolets Atomic Submarine", Woen, synthetic analog of thymalin, on the development of esopha Med. ZH, 9:28-33 (Medline Abstract No. 92101433). Khmelnitskii et al. (1991) "Morphofunctional Characteris gus and forestomach tumors induced by N-nitrososarcosine tics of the Immunocompetent System in Hypotrophy and Its ethyl ester in rats” Chem. Abstracts 111:23 Abstract No. Correction By Thymogen". Arkh. Patol. 53:24-27 (Medline 146389r (1989). Abstract No. 92171772). Gee, N.S. and Kenny, A.J., "Proteins of the kidney microvil Low et al. (1981) Proceedings of the National Academy of lar membranes' Biochem. J. 246:97-102 (1987). Science, 78:1162-1166. Jaroff, L. "Stop that Germ!" Time, pp.55-64 (May 23, Rodionov et al., (1991) “Natural Killer Activity in Patients 1988). with Chronic Dermatoses". Vestn. Dernatol. Venerol. 5:4-6, Pettit, G.R. in: Synthetic Peptides, vol. 3. Academic Press, (Biological Abstract vol. 92, Abstract No. 100385). NY. pp. 130, 134, 143, 424, 427, 429, and 434 (1975). Rodionov et al. (1990) "The Immunocorrective Therapy of Remington's Pharmaceutical Sciences. Mack Publishing Pyoderma Caused by Staphylococci Multiply Resistant to Co., Easton, PA. pp. 996-997, 1465-1469, 1553–1557 Antibiotics", Vestn. Dermatol. Wenerol. 1:42-45 (Medline (1980). Abstract No. 90224329). Sandström, E.G. and Kaplan, J.C.. "Antiviral therapy in Werner, G. H. et al. (1986) "Immunomodulating peptides." AIDS: Clinical pharmacological properties and therapeutic Experientia, 42:521-531. experience to date" Drugs 34:372-390 (1987). Aliev, M.G. et al., "Simulation of thymus dysfunction in Sievertsson, H. et al., "Synthesis of Di-and tripeptides and guinea pigs by using immunomodulators” Chem. Abstracts assay in Vivo for activity in the thyrotrol releasing hormone 116:664 Abstract No. 171986u (1992). and the luteinizing releasing hormone systems' J. Med. ASM News 56:368 (1990). Chem. 15(1):8-11 (1971). U.S. Patent Mar. 17, 1998 Sheet 1 of 8 5,728,680 L-Glu-L-Trp A/6 /4 - - 0 00 as 80 es a 60 40 & 20 O 0 0 20 30 40 50 60 0 80 TIME (HRS.) L-Glu-L-Trp + AMPICILLIN A/G As -- 0 -O-- 0 -O-- 00 -V- 000 O O 20 30 40 50 60 70 80 TIME (HRS.) U.S. Patent Mar. 17, 1998 Sheet 2 of 8 5,728,680 HRS. POST-CHALLENGE -- 0 A/6 WC -O-24 90 III, III 80 |A || || III 60 M II 50 |M|| || || || 40 MII-PH 30 A II AH-1H Us1H III e1 - O 00 l,000 DOSE (mcg/kg) L-Glu-L-Trp HRS. POST-CHALLENGE -- 0 -O- 24 -O- 48 - V - 72 p-Ho IIM III || || |A || || 1 |M|| ||U21|| AAI-21 U11 II || ||II /1 || || || A 1 ZY | | | | || A III O 00 1,000 DOSE (mcg/kg) L-Gly-L-Trp-AMPICILLIN U.S. Patent Mar. 17, 1998 Sheet 3 of 8 5,728,680 A/G 24 tl." s e s s t : A O IO 20 30 40 50 60 70 80 90 100 TIME (HRS) A/G 2A L-Glu-L-Trp--GENTAMYCIN 0 0 20 30 40 50 60 70 80 90 00 TIME (HRS.) U.S. Patent Mar. 17, 1998 Sheet 4 of 8 5,728,680 HRs POST-CHA.EGE -- 24 A/G 2C -O- 48 90 ---, 72 - 1||||IH || 1I - 50 1I-111T - II THIII ulti-HiIII III O 1II II 0. O 00 DOSE (mcg/kg) L-Glu-L-Trp HRS. POST - CHALLENGE - - 24 A/6 2 -O-96-O- 48 U-1 8 O III. 6 O 11|| 40 M1M || || 2O MM|| || || 0 MI, III100 1,000 DOSE (mcg/kg) L-Glu-L-Trp- GENTANYCIN U.S. Patent Mar. 17, 1998 Sheet 5 of 8 5,728,680 A/G 34 L-Glu--Tr"" -O--0. s 5CV s s 0 0 20 30 40 50 60 70 80 TIME (HRS) A/G 3A s 2 S s O IO 20 30 40 50 60 70 80 TIME (R.S.) U.S. Patent Mar. 17, 1998 Sheet 6 of 8 5,728,680 HRS. POST-CHALLENGE -- 24 A/G 3C -O- 48 -O-96 00 90 U-1H-1H 80 O - I 60 || | | | | || 50 III al-III 40 |A || ||U 30 A S1 20 LM-1 || Oc 4-1 I TTTI,O III100 I IIIl,000 DOSE (mcg/kg) L-Glu-L-Trp HRS.
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