|H|||||||||| USOO5278143A United States Patent 19 11 Patent Number: 5,278,143 Shepro et al. (45) Date of Patent: Jan. 11, 1994 (54) PROPHYLACTIC AND THERAPEUTIC OTHER PUBLICATIONS METHODS FOR TREATING NTER LEUKIN-MEDIATED EDEMAS Rudinger, Peptide Hormones, Parsons (Ed.) U. Park t Press, Baltimore, pp. 1-7 (1976). (75) Inventors: David Shepro, Boston, Mass.; J. Doukas et al., Blood, vol. 69, No. 6 pp. 1563-1569 (Jun. Steven Alexander, Nashville, Tenn. 1987). Frimmer, Chem. Abstracts, vol. 71, No. 99937m (1969). 73) Assignee: Trustees of Boston University, Wieland et al., Crit. Rev. Biochem. vol. 5, pp. 185-260 Boston, Mass. (1978). (21) Appl. No.: 807,668 Welbournet al., J. Appl. Physiol 70: 1364-1368 (1991). 22 illed: Dec. 16, 1991 Primary Examiner-Y. Christina Chan 22 Filed ec. 10, Attorney, Agent, or Firm-David Prashker Related U.S. Application Data 57 ABSTRACT m 63 continuation of ser. No. 416,905, Oct. 4, 1989, aban- Unique methods for treating interleukin-mediated ede doned, which is a continuation-in-part of Ser. No. mas in living subjects are provided comprising adminis 47,121, Oct. 4, 1989, abandoned, and a continuation- tering an effective amount of a composition selected in-part of Ser. No. 185,650, Apr. 25, 1988, abandoned. from the group consisting of phallotoxins, phallotoxin analogues, antamanide, or an antananide analogue to 51 Int. Cl. ........................ A61K 37/02; CK /6 the subject. The methods offer prophylactic and thera 52 576.5/35567; 17,353 peutic modes of treatment for both localized and sys s s 8 temic interleukin-mediated edemas. The compositions 58) Field of Search .................... 530/317,321; 514/9, of choice may be applied topically or given parenter 514/11, 870, 922; 424/85.2 ally; and may be employed with other diverse agents for (56) References Cited treatment of both inflammatory and non-inflammatory edemas. U.S. PATENT DOCUMENTS 4,690,915 9/1987 Rosenberg .............................. 514/2 7 Claims, 3 Drawing Sheets U.S. Patent Jan. 11, 1994 Sheet 1 of 3 5,278,143 ** 3: SIG. OIFF. FROM CONTROL SALINE (P<0.5, P<0.0) RESPECTIVELY ) ava SIG. DIFF. FROM IL-2 106 U/M 1 (P<0.05, PCO,O RESPECTIVELY) E at m - - a de e C C UMd a UM (V) MD GMD N. a. N N N a N. Cn N. cn (N ad - - ON t C l - - ad 3 - A. a with P O sed - n - a 5 MD Mo CM) N EA. S SQ s -N Nup CN NN CN and a 22 - b A. a es Ee S2 U.S. Patent Jan. 11, 1994 Sheet 2 of 3 5,278,143 3,3:3: SIG. DIFF. FRON CONTROL SALINE (P<0.05, P & O. O. RESPECTIVELY ) -- ~ SIC. DIFF. FRON IL-2 IO 6U/M (P<0.05, PCOO RESPECTIVELY i FIG. 3 i F G. 4. U.S. Patent Jan. 11, 1994 Sheet 3 of 3 5,278,143 %,3+ SIG. DIFF. FROM CONTROL SALINE (P<0.05, PCOO RESPECTIVELY ) ava SIG. Diff. FROM IL-20 6U/M (P<0.05, P&O.O RESPECTIVELY ) 5 a r 4 s 2250 2000 750 500 250 000 d ae CSS C S CC C C C CC S.C 750 C C S. O C C C C3 CC SS C 500 O C C SSC C 250 C 33. C 3. 3. SALNE SNE pRobinS SSs froE CONTROL IL-2 L-2 IL-2 FIG.6 TREATMENT 5,278,143 1 2 ter "nm') in radius or by slits about 8 nm wide. There is PROPHYLACTIC AND THERAPEUTICMETHODS also believed to be a system of larger sized pores about FOR TREATING INTERLEUKIN-MEDIATED 25 nm in radius which accounts for the small quantities EDEMAS of protein and other large solutes that normally cross the endothelial wall barrier. GOVERNMENTAL SUPPORT A variety of different disturbances can induce a con Research support for the present invention was pro dition of edema. These include: an elevated venous vided by a grant from the Community Technology hydrostatic pressure which may be caused by thrombo Foundation of Boston, University. sis of a vein or any other venous obstruction; hypopro This application is a continuation of application Ser. 10 teinemia with reduced plasma oncotic pressure result No. 416,905, filed Oct. 4, 1989, now abandoned; which ing from either inadequate synthesis or increased loss of is a continuation-in-part of U.S. patent application Ser. albumin; increased osmotic pressure of the interstitial No. 185,650 filed Apr. 25, 1988 now abandoned; and a fluid due to abnormal accumulation of sodium in the continuation-in-part of copending U.S. patent applica body because renal excretion of sodium cannot keep tion Ser. No. 417,121, filed Oct. 4, 1989, now aban 5 pace with the intake; failure of the lymphatics to re doned. move fluid and protein adequately from the interstitial space; an increased capillary permeability to fluids and FIELD OF THE INVENTION proteins as occurs in the inflammatory response to tissue The present invention is concerned with inflamma injury; an increased mucopolysaccharide content tory and non-inflammatory (hydraulic) associated ede 20 within the interstitial spaces; and an iatrogennic induced mas and is particularly directed to prophylactic and or mediated increase or accumulation of fluid and pro therapeutic methods for treating localized and systemic tein resulting from the administration of a pharmacolog edemas caused or mediated by interleukins. ically active lymphokine during the course of treatment BACKGROUND OF THE INVENTION by a physician or surgeon. This last-identified distur 25 bance merits a more detailed examination in view of Edema is the term generally used to describe the recent developments in immunotherapy. accumulation of excess fluid in the intercellular (inter Lymphokines comprise a broad class of biologically stitial) tissue spaces or body cavities. Edema may occur active substances which are secreted by various types of as a localized phenomenon such as the swelling of a leg lymphocytes in-vivo and in-vitro, especially by differ. when the venous outflow is obstructed; or it may be 30 ent populations of T-cell lymphocytes. Perhaps the systemic as in congestive heart failure or renal failure. When edema is severe and generalized, there is diffuse most controversial presently is the family of interleu swelling of all tissues and organs in the body and partic kins, which presently comprises: Interleukin-1 (“IL-1") ularly pronounced areas are given their own individual involved in the activation of resting T-cells; Interleukin names. For example, collection of edema in the perito 35 2 ("IL-2') which mediates the proliferation of T-cells neal cavity is known as "ascites'; accumulations of fluid and induces cytotoxic activity in T-cells; Interleukin-3 in the pleural cavity are termed "hydrothorax'; and ("IL-3') which causes the proliferation of mast cells edema of the pericardial sac is termed "pericardial effu and granulocytes; Interleukin-4 ("IL-4') which medi sion' or "hydropericardium'. Non-inflammatory ates the proliferation, activation, and differentiation of edema fluid such as accumulates in heart failure and B-cells, T-cells, and natural killer cells; and Interleukin renal disease is protein poor and referred to as a "transu 6 ("IL-6”) which induces the growth and differentiation date'. In contrast, inflammatory edema related to in of B-cells and T-cells Spits et al., J. Immunol. 139:1142 creased endothelial permeability is protein rich and is (1987); Kawakami et al., J. Exp. Med 168:2183 (1988); caused by the escape of plasma proteins (principally Spits et al., J. Immunol. 141:29 (1988); and Tartakovsky albumin) and polymorphonuclear leukocytes (hereinaf 45 et al., J. Immunol. 141:3863 (1983)). ter "PMNs') to form an exudate. Of particular interest is Interleukin-2 ("IL-2') and its Edena, whether inflammatory or non-inflammatory analogues whose isolation, chemical formulation and in nature, is thus an abnormality in the fluid balance structure, and synthesis by conventional wet chemis within the microcirculation which includes the small tries and by recombinant DNA techniques have been arterioles, capillaries, and post-capillary venules of the 50 intensively pursued see for example U.S. Pat. Nos. circulatory system. Normal fluid balance and exchange 4,490,289; 4,138,927; 4,569,790; 4,578,335; 4,761,375; is critically dependent on the presence of an intact and 4,518,584; 4,604,377; 4,564,593; and the references cited metabolically active endothelium within the vascula therein). The use of Interleukin-2 as a therapeutic com ture. Normal endothelium is a thin, squamous epit position has followed two very different approaches. belium adapted to permit free, rapid exchange of water 55 The first line of development is exemplified by the IL-2 and small molecules between plasma and interstitium; immunotherapy technique for treating cancers origi but one which limits the passage of plasma proteins with nated by Dr. Steven A. Rosenberg Rosenberg, S.A., increases in protein size. Immuno. Today 9:58-62 (1988) and the references cited The endothelial lining of all arterioles and venules, therein). That approach involves the removal of lym and most capillaries in the body, is of the continuous phoid cells from a tumor-bearing host; an in-vitro ex type, having an unbroken cytoplasmic layer with pansion of the host's cells by culture in Interleukin-2 closely apposed intercellular junctions. Physiological containing culture media to produce lymphokine studies Renkin, E., Circ. Res. 41:735-743 (1977); Ren activated killer ("LAK') cells; and a re-introduction of kin, E., ACTA Physiol. Scand. (Suppl.) 463:81 (1979); the LAK cell culture into the living host accompanied Bottaro et al., Microvasc.